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Sample records for nostoc sp pcc

  1. Arsenic biotransformation by a cyanobacterium Nostoc sp. PCC 7120.

    Science.gov (United States)

    Xue, Xi-Mei; Yan, Yu; Xiong, Chan; Raber, Georg; Francesconi, Kevin; Pan, Ting; Ye, Jun; Zhu, Yong-Guan

    2017-09-01

    Nostoc sp. PCC 7120 (Nostoc), a typical filamentous cyanobacterium ubiquitous in aquatic system, is recognized as a model organism to study prokaryotic cell differentiation and nitrogen fixation. In this study, Nostoc cells incubated with arsenite (As(III)) for two weeks were extracted with dichloromethane/methanol (DCM/MeOH) and the extract was partitioned between water and DCM. Arsenic species in aqueous and DCM layers were determined using high performance liquid chromatography - inductively coupled plasma mass spectrometer/electrospray tandem mass spectrometry (HPLC-ICPMS/ESIMSMS). In addition to inorganic arsenic (iAs), the aqueous layer also contained monomethylarsonate (MAs(V)), dimethylarsinate (DMAs(V)), and the two arsenosugars, namely a glycerol arsenosugar (Oxo-Gly) and a phosphate arsenosugar (Oxo-PO4). Two major arsenosugar phospholipids (AsSugPL982 and AsSugPL984) were detected in DCM fraction. Arsenic in the growth medium was also investigated by HPLC/ICPMS and shown to be present mainly as the inorganic forms As(III) and As(V) accounting for 29%-38% and 29%-57% of the total arsenic respectively. The total arsenic of methylated arsenic, arsenosugars, and arsenosugar phospholipids in Nostoc cells with increasing As(III) exposure were not markedly different, indicating that the transformation to organoarsenic in Nostoc was not dependent on As(III) concentration in the medium. Our results provide new insights into the role of cyanobacteria in the biogeochemical cycling of arsenic. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Transcription and Regulation of the Bidirectional Hydrogenase in the Cyanobacterium Nostoc sp. Strain PCC 7120▿

    Science.gov (United States)

    Sjöholm, Johannes; Oliveira, Paulo; Lindblad, Peter

    2007-01-01

    The filamentous, heterocystous cyanobacterium Nostoc sp. strain PCC 7120 (Anabaena sp. strain PCC 7120) possesses an uptake hydrogenase and a bidirectional enzyme, the latter being capable of catalyzing both H2 production and evolution. The completely sequenced genome of Nostoc sp. strain PCC 7120 reveals that the five structural genes encoding the bidirectional hydrogenase (hoxEFUYH) are separated in two clusters at a distance of approximately 8.8 kb. The transcription of the hox genes was examined under nitrogen-fixing conditions, and the results demonstrate that the cluster containing hoxE and hoxF can be transcribed as one polycistronic unit together with the open reading frame alr0750. The second cluster, containing hoxU, hoxY, and hoxH, is transcribed together with alr0763 and alr0765, located between the hox genes. Moreover, alr0760 and alr0761 form an additional larger operon. Nevertheless, Northern blot hybridizations revealed a rather complex transcription pattern in which the different hox genes are expressed differently. Transcriptional start points (TSPs) were identified 66 and 57 bp upstream from the start codon of alr0750 and hoxU, respectively. The transcriptions of the two clusters containing the hox genes are both induced under anaerobic conditions concomitantly with the induction of a higher level of hydrogenase activity. An additional TSP, within the annotated alr0760, 244 bp downstream from the suggested translation start codon, was identified. Electrophoretic mobility shift assays with purified LexA from Nostoc sp. strain PCC 7120 demonstrated specific interactions between the transcriptional regulator and both hox promoter regions. However, when LexA from Synechocystis sp. strain PCC 6803 was used, the purified protein interacted only with the promoter region of the alr0750-hoxE-hoxF operon. A search of the whole Nostoc sp. strain PCC 7120 genome demonstrated the presence of 216 putative LexA binding sites in total, including recA and rec

  3. Transcription of the extended hyp-operon in Nostoc sp. strain PCC 7120

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    Lindblad Peter

    2008-04-01

    Full Text Available Abstract Background The maturation of hydrogenases into active enzymes is a complex process and e.g. a correctly assembled active site requires the involvement of at least seven proteins, encoded by hypABCDEF and a hydrogenase specific protease, encoded either by hupW or hoxW. The N2-fixing cyanobacterium Nostoc sp. strain PCC 7120 may contain both an uptake and a bidirectional hydrogenase. The present study addresses the presence and expression of hyp-genes in Nostoc sp. strain PCC 7120. Results RT-PCRs demonstrated that the six hyp-genes together with one ORF may be transcribed as a single operon. Transcriptional start points (TSPs were identified 280 bp upstream from hypF and 445 bp upstream of hypC, respectively, demonstrating the existence of several transcripts. In addition, five upstream ORFs located in between hupSL, encoding the small and large subunits of the uptake hydrogenase, and the hyp-operon, and two downstream ORFs from the hyp-genes were shown to be part of the same transcript unit. A third TSP was identified 45 bp upstream of asr0689, the first of five ORFs in this operon. The ORFs are annotated as encoding unknown proteins, with the exception of alr0692 which is identified as a NifU-like protein. Orthologues of the four ORFs asr0689-alr0692, with a highly conserved genomic arrangement positioned between hupSL, and the hyp genes are found in several other N2-fixing cyanobacteria, but are absent in non N2-fixing cyanobacteria with only the bidirectional hydrogenase. Short conserved sequences were found in six intergenic regions of the extended hyp-operon, appearing between 11 and 79 times in the genome. Conclusion This study demonstrated that five ORFs upstream of the hyp-gene cluster are co-transcribed with the hyp-genes, and identified three TSPs in the extended hyp-gene cluster in Nostoc sp. strain PCC 7120. This may indicate a function related to the assembly of a functional uptake hydrogenase, hypothetically in the

  4. Transcription of the extended hyp-operon in Nostoc sp. strain PCC 7120

    Science.gov (United States)

    Agervald, Åsa; Stensjö, Karin; Holmqvist, Marie; Lindblad, Peter

    2008-01-01

    Background The maturation of hydrogenases into active enzymes is a complex process and e.g. a correctly assembled active site requires the involvement of at least seven proteins, encoded by hypABCDEF and a hydrogenase specific protease, encoded either by hupW or hoxW. The N2-fixing cyanobacterium Nostoc sp. strain PCC 7120 may contain both an uptake and a bidirectional hydrogenase. The present study addresses the presence and expression of hyp-genes in Nostoc sp. strain PCC 7120. Results RT-PCRs demonstrated that the six hyp-genes together with one ORF may be transcribed as a single operon. Transcriptional start points (TSPs) were identified 280 bp upstream from hypF and 445 bp upstream of hypC, respectively, demonstrating the existence of several transcripts. In addition, five upstream ORFs located in between hupSL, encoding the small and large subunits of the uptake hydrogenase, and the hyp-operon, and two downstream ORFs from the hyp-genes were shown to be part of the same transcript unit. A third TSP was identified 45 bp upstream of asr0689, the first of five ORFs in this operon. The ORFs are annotated as encoding unknown proteins, with the exception of alr0692 which is identified as a NifU-like protein. Orthologues of the four ORFs asr0689-alr0692, with a highly conserved genomic arrangement positioned between hupSL, and the hyp genes are found in several other N2-fixing cyanobacteria, but are absent in non N2-fixing cyanobacteria with only the bidirectional hydrogenase. Short conserved sequences were found in six intergenic regions of the extended hyp-operon, appearing between 11 and 79 times in the genome. Conclusion This study demonstrated that five ORFs upstream of the hyp-gene cluster are co-transcribed with the hyp-genes, and identified three TSPs in the extended hyp-gene cluster in Nostoc sp. strain PCC 7120. This may indicate a function related to the assembly of a functional uptake hydrogenase, hypothetically in the assembly of the small subunit of

  5. BMAA Inhibits Nitrogen Fixation in the Cyanobacterium Nostoc sp. PCC 7120

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    Birgitta Bergman

    2013-08-01

    Full Text Available Cyanobacteria produce a range of secondary metabolites, one being the neurotoxic non-protein amino acid β-N-methylamino-L-alanine (BMAA, proposed to be a causative agent of human neurodegeneration. As for most cyanotoxins, the function of BMAA in cyanobacteria is unknown. Here, we examined the effects of BMAA on the physiology of the filamentous nitrogen-fixing cyanobacterium Nostoc sp. PCC 7120. Our data show that exogenously applied BMAA rapidly inhibits nitrogenase activity (acetylene reduction assay, even at micromolar concentrations, and that the inhibition was considerably more severe than that induced by combined nitrogen sources and most other amino acids. BMAA also caused growth arrest and massive cellular glycogen accumulation, as observed by electron microscopy. With nitrogen fixation being a process highly sensitive to oxygen species we propose that the BMAA effects found here may be related to the production of reactive oxygen species, as reported for other organisms.

  6. BMAA Inhibits Nitrogen Fixation in the Cyanobacterium Nostoc sp. PCC 7120

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    Berntzon, Lotta; Erasmie, Sven; Celepli, Narin; Eriksson, Johan; Rasmussen, Ulla; Bergman, Birgitta

    2013-01-01

    Cyanobacteria produce a range of secondary metabolites, one being the neurotoxic non-protein amino acid β-N-methylamino-L-alanine (BMAA), proposed to be a causative agent of human neurodegeneration. As for most cyanotoxins, the function of BMAA in cyanobacteria is unknown. Here, we examined the effects of BMAA on the physiology of the filamentous nitrogen-fixing cyanobacterium Nostoc sp. PCC 7120. Our data show that exogenously applied BMAA rapidly inhibits nitrogenase activity (acetylene reduction assay), even at micromolar concentrations, and that the inhibition was considerably more severe than that induced by combined nitrogen sources and most other amino acids. BMAA also caused growth arrest and massive cellular glycogen accumulation, as observed by electron microscopy. With nitrogen fixation being a process highly sensitive to oxygen species we propose that the BMAA effects found here may be related to the production of reactive oxygen species, as reported for other organisms. PMID:23966039

  7. Arsenic Demethylation by a C·As Lyase in Cyanobacterium Nostoc sp. PCC 7120.

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    Yan, Yu; Ye, Jun; Xue, Xi-Mei; Zhu, Yong-Guan

    2015-12-15

    Arsenic, a ubiquitous toxic substance, exists mainly as inorganic forms in the environment. It is perceived that organoarsenicals can be demethylated and degraded into inorganic arsenic by microorganisms. Few studies have focused on the mechanism of arsenic demethylation in bacteria. Here, we investigated arsenic demethylation in a typical freshwater cyanobacterium Nostoc sp. PCC 7120. This bacterium was able to demethylate monomethylarsenite [MAs(III)] rapidly to arsenite [As(III)] and also had the ability to demethylate monomethylarsenate [MAs(V)] to As(III). The NsarsI encoding a C·As lyase responsible for MAs(III) demethylation was cloned from Nostoc sp. PCC 7120 and heterologously expressed in an As-hypersensitive strain Escherichia coli AW3110 (ΔarsRBC). Expression of NsarsI was shown to confer MAs(III) resistance through arsenic demethylation. The purified NsArsI was further identified and functionally characterized in vitro. NsArsI existed mainly as the trimeric state, and the kinetic data were well-fit to the Hill equation with K0.5 = 7.55 ± 0.33 μM for MAs(III), Vmax = 0.79 ± 0.02 μM min(-1), and h = 2.7. Both of the NsArsI truncated derivatives lacking the C-terminal 10 residues (ArsI10) or 23 residues (ArsI23) had a reduced ability of MAs(III) demethylation. These results provide new insights for understanding the important role of cyanobacteria in arsenic biogeochemical cycling in the environment.

  8. Biochemical Characterization of Putative Adenylate Dimethylallyltransferase and Cytokinin Dehydrogenase from Nostoc sp. PCC 7120.

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    Frébortová, Jitka; Greplová, Marta; Seidl, Michael F; Heyl, Alexander; Frébort, Ivo

    2015-01-01

    Cytokinins, a class of phytohormones, are adenine derivatives common to many different organisms. In plants, these play a crucial role as regulators of plant development and the reaction to abiotic and biotic stress. Key enzymes in the cytokinin synthesis and degradation in modern land plants are the isopentyl transferases and the cytokinin dehydrogenases, respectively. Their encoding genes have been probably introduced into the plant lineage during the primary endosymbiosis. To shed light on the evolution of these proteins, the genes homologous to plant adenylate isopentenyl transferase and cytokinin dehydrogenase were amplified from the genomic DNA of cyanobacterium Nostoc sp. PCC 7120 and expressed in Escherichia coli. The putative isopentenyl transferase was shown to be functional in a biochemical assay. In contrast, no enzymatic activity was detected for the putative cytokinin dehydrogenase, even though the principal domains necessary for its function are present. Several mutant variants, in which conserved amino acids in land plant cytokinin dehydrogenases had been restored, were inactive. A combination of experimental data with phylogenetic analysis indicates that adenylate-type isopentenyl transferases might have evolved several times independently. While the Nostoc genome contains a gene coding for protein with characteristics of cytokinin dehydrogenase, the organism is not able to break down cytokinins in the way shown for land plants.

  9. Near infrared fluorescent biliproteins generated from bacteriophytochrome AphB of Nostoc sp. PCC 7120.

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    Yuan, Che; Li, Hui-Zhen; Tang, Kun; Gärtner, Wolfgang; Scheer, Hugo; Zhou, Ming; Zhao, Kai-Hong

    2016-04-01

    The genome of the cyanobacterium Nostoc sp. PCC 7120 encodes a large number of putative bacteriophytochrome and cyanobacteriochrome photoreceptors that, due to their long-wavelength absorption and fluorescence emission, might serve as fluorescent tags in intracellular investigations. We show that the PAS-GAF domain of the bacteriophytochrome, AphB, binds biliverdin covalently and exhibits, besides its reversible photochemistry, a moderate fluorescence in the near infrared (NIR) spectral region. It was selected for further increasing the brightness while retaining the NIR fluorescence. In the first step, amino acids assumed to improve fluorescence were selectively mutated. The resulting variants were then subjected to several rounds of random mutagenesis and screened for enhanced fluorescence in the NIR. The brightness of optimized PAS-GAF variants increased more than threefold compared to that of wt AphB(1-321), with only insignificant spectral shifts (Amax around 695 nm, and Fmax around 720 nm). In general, the brightness increases with decreasing wavelengths, which allows for a selection of the fluorophore depending on the optical properties of the tissue. A spectral heterogeneity was observed when residue His260, located in close proximity to the chromophore, was mutated to Tyr, emphasizing the strong effects of the environment on the electronic properties of the bound biliverdin chromophore.

  10. Diversity and transcription of proteases involved in the maturation of hydrogenases in Nostoc punctiforme ATCC 29133 and Nostoc sp. strain PCC 7120

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    2009-01-01

    Background The last step in the maturation process of the large subunit of [NiFe]-hydrogenases is a proteolytic cleavage of the C-terminal by a hydrogenase specific protease. Contrary to other accessory proteins these hydrogenase proteases are believed to be specific whereby one type of hydrogenases specific protease only cleaves one type of hydrogenase. In cyanobacteria this is achieved by the gene product of either hupW or hoxW, specific for the uptake or the bidirectional hydrogenase respectively. The filamentous cyanobacteria Nostoc punctiforme ATCC 29133 and Nostoc sp strain PCC 7120 may contain a single uptake hydrogenase or both an uptake and a bidirectional hydrogenase respectively. Results In order to examine these proteases in cyanobacteria, transcriptional analyses were performed of hupW in Nostoc punctiforme ATCC 29133 and hupW and hoxW in Nostoc sp. strain PCC 7120. These studies revealed numerous transcriptional start points together with putative binding sites for NtcA (hupW) and LexA (hoxW). In order to investigate the diversity and specificity among hydrogeanse specific proteases we constructed a phylogenetic tree which revealed several subgroups that showed a striking resemblance to the subgroups previously described for [NiFe]-hydrogenases. Additionally the proteases specificity was also addressed by amino acid sequence analysis and protein-protein docking experiments with 3D-models derived from bioinformatic studies. These studies revealed a so called "HOXBOX"; an amino acid sequence specific for protease of Hox-type which might be involved in docking with the large subunit of the hydrogenase. Conclusion Our findings suggest that the hydrogenase specific proteases are under similar regulatory control as the hydrogenases they cleave. The result from the phylogenetic study also indicates that the hydrogenase and the protease have co-evolved since ancient time and suggests that at least one major horizontal gene transfer has occurred. This co

  11. Diversity and transcription of proteases involved in the maturation of hydrogenases in Nostoc punctiforme ATCC 29133 and Nostoc sp. strain PCC 7120

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    Lindblad Peter

    2009-03-01

    Full Text Available Abstract Background The last step in the maturation process of the large subunit of [NiFe]-hydrogenases is a proteolytic cleavage of the C-terminal by a hydrogenase specific protease. Contrary to other accessory proteins these hydrogenase proteases are believed to be specific whereby one type of hydrogenases specific protease only cleaves one type of hydrogenase. In cyanobacteria this is achieved by the gene product of either hupW or hoxW, specific for the uptake or the bidirectional hydrogenase respectively. The filamentous cyanobacteria Nostoc punctiforme ATCC 29133 and Nostoc sp strain PCC 7120 may contain a single uptake hydrogenase or both an uptake and a bidirectional hydrogenase respectively. Results In order to examine these proteases in cyanobacteria, transcriptional analyses were performed of hupW in Nostoc punctiforme ATCC 29133 and hupW and hoxW in Nostoc sp. strain PCC 7120. These studies revealed numerous transcriptional start points together with putative binding sites for NtcA (hupW and LexA (hoxW. In order to investigate the diversity and specificity among hydrogeanse specific proteases we constructed a phylogenetic tree which revealed several subgroups that showed a striking resemblance to the subgroups previously described for [NiFe]-hydrogenases. Additionally the proteases specificity was also addressed by amino acid sequence analysis and protein-protein docking experiments with 3D-models derived from bioinformatic studies. These studies revealed a so called "HOXBOX"; an amino acid sequence specific for protease of Hox-type which might be involved in docking with the large subunit of the hydrogenase. Conclusion Our findings suggest that the hydrogenase specific proteases are under similar regulatory control as the hydrogenases they cleave. The result from the phylogenetic study also indicates that the hydrogenase and the protease have co-evolved since ancient time and suggests that at least one major horizontal gene transfer

  12. Structure-Function, Stability, and Chemical Modification of the Cyanobacterial Cytochrome b6f Complex from Nostoc sp. PCC 7120*

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    Baniulis, Danas; Yamashita, Eiki; Whitelegge, Julian P.; Zatsman, Anna I.; Hendrich, Michael P.; Hasan, S. Saif; Ryan, Christopher M.; Cramer, William A.

    2009-01-01

    The crystal structure of the cyanobacterial cytochrome b6f complex has previously been solved to 3.0-Å resolution using the thermophilic Mastigocladus laminosus whose genome has not been sequenced. Several unicellular cyanobacteria, whose genomes have been sequenced and are tractable for mutagenesis, do not yield b6f complex in an intact dimeric state with significant electron transport activity. The genome of Nostoc sp. PCC 7120 has been sequenced and is closer phylogenetically to M. laminosus than are unicellular cyanobacteria. The amino acid sequences of the large core subunits and four small peripheral subunits of Nostoc are 88 and 80% identical to those in the M. laminosus b6f complex. Purified b6f complex from Nostoc has a stable dimeric structure, eight subunits with masses similar to those of M. laminosus, and comparable electron transport activity. The crystal structure of the native b6f complex, determined to a resolution of 3.0Å (PDB id: 2ZT9), is almost identical to that of M. laminosus. Two unique aspects of the Nostoc complex are: (i) a dominant conformation of heme bp that is rotated 180° about the α- and γ-meso carbon axis relative to the orientation in the M. laminosus complex and (ii) acetylation of the Rieske iron-sulfur protein (PetC) at the N terminus, a post-translational modification unprecedented in cyanobacterial membrane and electron transport proteins, and in polypeptides of cytochrome bc complexes from any source. The high spin electronic character of the unique heme cn is similar to that previously found in the b6f complex from other sources. PMID:19189962

  13. Transcript analysis of the extended hyp-operon in the cyanobacteria Nostoc sp. strain PCC 7120 and Nostoc punctiforme ATCC 29133

    Science.gov (United States)

    2011-01-01

    Background Cyanobacteria harbor two [NiFe]-type hydrogenases consisting of a large and a small subunit, the Hup- and Hox-hydrogenase, respectively. Insertion of ligands and correct folding of nickel-iron hydrogenases require assistance of accessory maturation proteins (encoded by the hyp-genes). The intergenic region between the structural genes encoding the uptake hydrogenase (hupSL) and the accessory maturation proteins (hyp genes) in the cyanobacteria Nostoc PCC 7120 and N. punctiforme were analysed using molecular methods. Findings The five ORFs, located in between the uptake hydrogenase structural genes and the hyp-genes, can form a transcript with the hyp-genes. An identical genomic localization of these ORFs are found in other filamentous, N2-fixing cyanobacterial strains. In N. punctiforme and Nostoc PCC 7120 the ORFs upstream of the hyp-genes showed similar transcript level profiles as hupS (hydrogenase structural gene), nifD (nitrogenase structural gene), hypC and hypF (accessory hydrogenase maturation genes) after nitrogen depletion. In silico analyzes showed that these ORFs in N. punctiforme harbor the same conserved regions as their homologues in Nostoc PCC 7120 and that they, like their homologues in Nostoc PCC 7120, can be transcribed together with the hyp-genes forming a larger extended hyp-operon. DNA binding studies showed interactions of the transcriptional regulators CalA and CalB to the promoter regions of the extended hyp-operon in N. punctiforme and Nostoc PCC 7120. Conclusions The five ORFs upstream of the hyp-genes in several filamentous N2-fixing cyanobacteria have an identical genomic localization, in between the genes encoding the uptake hydrogenase and the maturation protein genes. In N. punctiforme and Nostoc PCC 7120 they are transcribed as one operon and may form transcripts together with the hyp-genes. The expression pattern of the five ORFs within the extended hyp-operon in both Nostoc punctiforme and Nostoc PCC 7120 is similar to

  14. Identification and characterization of a carboxysomal γ-carbonic anhydrase from the cyanobacterium Nostoc sp. PCC 7120.

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    de Araujo, Charlotte; Arefeen, Dewan; Tadesse, Yohannes; Long, Benedict M; Price, G Dean; Rowlett, Roger S; Kimber, Matthew S; Espie, George S

    2014-09-01

    Carboxysomes are proteinaceous microcompartments that encapsulate carbonic anhydrase (CA) and ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco); carboxysomes, therefore, catalyze reversible HCO3 (-) dehydration and the subsequent fixation of CO2. The N- and C-terminal domains of the β-carboxysome scaffold protein CcmM participate in a network of protein-protein interactions that are essential for carboxysome biogenesis, organization, and function. The N-terminal domain of CcmM in the thermophile Thermosynechococcus elongatus BP-1 is also a catalytically active, redox regulated γ-CA. To experimentally determine if CcmM from a mesophilic cyanobacterium is active, we cloned, expressed and purified recombinant, full-length CcmM from Nostoc sp. PCC 7120 as well as the N-terminal 209 amino acid γ-CA-like domain. Both recombinant proteins displayed ethoxyzolamide-sensitive CA activity in mass spectrometric assays, as did the carboxysome-enriched TP fraction. NstCcmM209 was characterized as a moderately active and efficient γ-CA with a k cat of 2.0 × 10(4) s(-1) and k cat/K m of 4.1 × 10(6) M(-1) s(-1) at 25 °C and pH 8, a pH optimum between 8 and 9.5 and a temperature optimum spanning 25-35 °C. NstCcmM209 also catalyzed the hydrolysis of the CO2 analog carbonyl sulfide. Circular dichroism and intrinsic tryptophan fluorescence analysis demonstrated that NstCcmM209 was progressively and irreversibly denatured above 50 °C. NstCcmM209 activity was inhibited by the reducing agent tris(hydroxymethyl)phosphine, an effect that was fully reversed by a molar excess of diamide, a thiol oxidizing agent, consistent with oxidative activation being a universal regulatory mechanism of CcmM orthologs. Immunogold electron microscopy and Western blot analysis of TP pellets indicated that Rubisco and CcmM co-localize and are concentrated in Nostoc sp. PCC 7120 carboxysomes.

  15. New Insight into the Cleavage Reaction of Nostoc sp. Strain PCC 7120 Carotenoid Cleavage Dioxygenase in Natural and Nonnatural Carotenoids

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    Heo, Jinsol; Kim, Se Hyeuk

    2013-01-01

    Carotenoid cleavage dioxygenases (CCDs) are enzymes that catalyze the oxidative cleavage of carotenoids at a specific double bond to generate apocarotenoids. In this study, we investigated the activity and substrate preferences of NSC3, a CCD of Nostoc sp. strain PCC 7120, in vivo and in vitro using natural and nonnatural carotenoid structures. NSC3 cleaved β-apo-8′-carotenal at 3 positions, C-13C-14, C-15C-15′, and C-13′C-14′, revealing a unique cleavage pattern. NSC3 cleaves the natural structure of carotenoids 4,4′-diaponeurosporene, 4,4′-diaponeurosporen-4′-al, 4,4′-diaponeurosporen-4′-oic acid, 4,4′-diapotorulene, and 4,4′-diapotorulen-4′-al to generate novel cleavage products (apo-14′-diaponeurosporenal, apo-13′-diaponeurosporenal, apo-10′-diaponeurosporenal, apo-14′-diapotorulenal, and apo-10′-diapotorulenal, respectively). The study of carotenoids with natural or nonnatural structures produced by using synthetic modules could provide information valuable for understanding the cleavage reactions or substrate preferences of other CCDs in vivo and in vitro. PMID:23524669

  16. Retinal is formed from apo-carotenoids in Nostoc sp. PCC7120: in vitro characterization of an apo-carotenoid oxygenase

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    Scherzinger, Daniel; Ruch, Sandra; Kloer, Daniel P.; Wilde, Annegret; Al-Babili, Salim

    2006-01-01

    The sensory rhodopsin from Anabaena (Nostoc) sp. PCC7120 is the first cyanobacterial retinylidene protein identified. Here, we report on NosACO (Nostoc apo-carotenoid oxygenase), encoded by the ORF (open reading frame) all4284, as the candidate responsible for the formation of the required chromophore, retinal. In contrast with the enzymes from animals, NosACO converts β-apo-carotenals instead of β-carotene into retinal in vitro. The identity of the enzymatic products was proven by HPLC and gas chromatography–MS. NosACO exhibits a wide substrate specificity with respect to chain lengths and functional end-groups, converting β-apo-carotenals, (3R)-3-hydroxy-β-apo-carotenals and the corresponding alcohols into retinal and (3R)-3-hydroxyretinal respectively. However, kinetic analyses revealed very divergent Km and Vmax values. On the basis of the crystal structure of SynACO (Synechocystis sp. PCC6803 apo-carotenoid oxygenase), a related enzyme showing similar enzymatic activity, we designed a homology model of the native NosACO. The deduced structure explains the absence of β-carotene-cleavage activity and indicates that NosACO is a monotopic membrane protein. Accordingly, NosACO could be readily reconstituted into liposomes. To localize SynACO in vivo, a Synechocystis knock-out strain was generated expressing SynACO as the sole carotenoid oxygenase. Western-blot analyses showed that the main portion of SynACO occurred in a membrane-bound form. PMID:16759173

  17. HupW Protease Specifically Required for Processing of the Catalytic Subunit of the Uptake Hydrogenase in the Cyanobacterium Nostoc sp. Strain PCC 7120

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    Lindberg, Pia; Devine, Ellenor; Stensjö, Karin

    2012-01-01

    The maturation process of [NiFe] hydrogenases includes a proteolytic cleavage of the large subunit. We constructed a mutant of Nostoc strain PCC 7120 in which hupW, encoding a putative hydrogenase-specific protease, is inactivated. Our results indicate that the protein product of hupW selectively cleaves the uptake hydrogenase in this cyanobacterium. PMID:22020512

  18. Colour evaluation of a phycobiliprotein-rich extract obtained from Nostoc PCC9205 in acidic solutions and yogurt.

    Science.gov (United States)

    de O Moreira, Isabela; Passos, Thaís S; Chiapinni, Claudete; Silveira, Gabrielle K; Souza, Joana C M; Coca-Vellarde, Luis Guillermo; Deliza, Rosires; de Lima Araújo, Kátia G

    2012-02-01

    Phycobiliproteins are coloured proteins produced by cyanobacteria, which have several applications because of their colour properties. However, there is no available information about the colour stability of phycobiliproteins from Nostoc sp. in food systems. The aim of this work was to study the colour stability of a purple-coloured phycobiliprotein-rich extract from the cyanobacterium Nostoc PCC9205 in acidic solutions and yogurt. Variations of pH for Nostoc PCC9205 extract have shown stability for the L* (lightness) and a* (redness) indexes in the range 1.0-7.0. The b* index (blueness), however, increased at pH values below 4.0, indicating loss of the blue colour. The Nostoc PCC9205 extract was used as colorant in yogurt (pH 4.17) stored for 60 days. Instrumental colour analysis showed no changes for the L* and a* indexes during storage, whereas the b* index changed after 20 days of storage. A multiple comparison test showed colour instability after 20 days of storage. A hedonic scale test performed on the 60th day of storage showed acceptability of the product. The red component of the phycobiliprotein-rich extract from Nostoc PCC9205 presented an improved stability in acidic media and yogurt compared with the blue component of this extract. Copyright © 2011 Society of Chemical Industry.

  19. Discovery of Rare and Highly Toxic Microcystins from Lichen-Associated Cyanobacterium Nostoc sp. Strain IO-102-I

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    Oksanen, Ilona; Jokela, Jouni; Fewer, David P.; Wahlsten, Matti; Rikkinen, Jouko; Sivonen, Kaarina

    2004-01-01

    The production of hepatotoxic cyclic heptapeptides, microcystins, is almost exclusively reported from planktonic cyanobacteria. Here we show that a terrestrial cyanobacterium Nostoc sp. strain IO-102-I isolated from a lichen association produces six different microcystins. Microcystins were identified with liquid chromatography-UV mass spectrometry by their retention times, UV spectra, mass fragmentation, and comparison to microcystins from the aquatic Nostoc sp. strain 152. The dominant microcystin produced by Nostoc sp. strain IO-102-I was the highly toxic [ADMAdda5]microcystin-LR, which accounted for ca. 80% of the total microcystins. We assigned a structure of [DMAdda5]microcystin-LR and [d-Asp3,ADMAdda5]microcystin-LR and a partial structure of three new [ADMAdda5]-XR type of microcystin variants. Interestingly, Nostoc spp. strains IO-102-I and 152 synthesized only the rare ADMAdda and DMAdda subfamilies of microcystin variants. Phylogenetic analyses demonstrated congruence between genes involved directly in microcystin biosynthesis and the 16S rRNA and rpoC1 genes of Nostoc sp. strain IO-102-I. Nostoc sp. strain 152 and the Nostoc sp. strain IO-102-I are distantly related, revealing a sporadic distribution of toxin production in the genus Nostoc. Nostoc sp. strain IO-102-I is closely related to Nostoc punctiforme PCC 73102 and other symbiotic Nostoc strains and most likely belongs to this species. Together, this suggests that other terrestrial and aquatic strains of the genus Nostoc may have retained the genes necessary for microcystin biosynthesis. PMID:15466511

  20. CalA, a Cyanobacterial AbrB Protein, Interacts with the Upstream Region of hypC and Acts as a Repressor of Its Transcription in the Cyanobacterium Nostoc sp. Strain PCC 7120▿ †

    Science.gov (United States)

    Agervald, Åsa; Zhang, Xiaohui; Stensjö, Karin; Devine, Ellenor; Lindblad, Peter

    2010-01-01

    The filamentous, heterocystous, nitrogen-fixing cyanobacterium Nostoc sp. strain PCC 7120 may contain, depending on growth conditions, up to two hydrogenases directly involved in hydrogen metabolism. HypC is one out of at least seven auxiliary gene products required for synthesis of a functional hydrogenase, specifically involved in the maturation of the large subunit. In this study we present a protein, CalA (Alr0946 in the genome), belonging to the transcription regulator family AbrB, which in protein-DNA assays was found to interact with the upstream region of hypC. Transcriptional investigations showed that calA is cotranscribed with the downstream gene alr0947, which encodes a putative protease from the abortive infection superfamily, Abi. CalA was shown to interact specifically not only with the upstream region of hypC but also with its own upstream region, acting as a repressor on hypC. The bidirectional hydrogenase activity was significantly downregulated when CalA was overexpressed, demonstrating a correlation with the transcription factor, either direct or indirect. In silico studies showed that homologues to both CalA and Alr0947 are highly conserved proteins within cyanobacteria with very similar physical organizations of the corresponding structural genes. Possible functions of the cotranscribed downstream protein Alr0947 are presented. In addition, we present a three-dimensional (3D) model of the DNA binding domain of CalA and putative DNA binding mechanisms are discussed. PMID:20023111

  1. Nostoc thermotolerans sp. nov., a soil-dwelling species of Nostoc (Cyanobacteria).

    Science.gov (United States)

    Suradkar, Archana; Villanueva, Chelsea; Gaysina, Lira A; Casamatta, Dale A; Saraf, Aniket; Dighe, Gandhali; Mergu, Ratnaprabha; Singh, Prashant

    2017-05-01

    A filamentous, soil-dwelling cyanobacterial strain (9C-PST) was isolated from Mandsaur, Madhya Pradesh, India, and is described as a new species of the genus Nostoc. Extensive morphological and molecular characterization along with a thorough assessment of ecology was performed. The style of filament orientation, type and nature of the sheath (e.g. distribution and visibility across the trichome), and vegetative and heterocyte cell dimensions and shape were assessed for over one year using both the laboratory grown culture and the naturally occurring samples. Sequencing of the 16S rRNA gene showed 94 % similarity with Nostocpiscinale CENA21 while analyses of the secondary structures of the 16S-23S ITS region showed unique folding patterns that differentiated this strain from other species of Nostoc. The level of rbcl and rpoC1 gene sequence similarity was 91 and 94 % to Nostocsp. PCC 7524 and Nostocpiscinale CENA21, respectively, while the nifD gene sequence similarity was found to be 99 % with Nostocpiscinale CENA21. The phenotypic, ecological, genetic and phylogenetic observations indicate that the strain 9C-PST represents a novel species of the genus Nostoc with the name proposed being Nostoc thermotolerans sp. nov. according to the International Code of Nomenclature for Algae, Fungi, and Plants.

  2. Nostoc PCC7524, a cyanobacterium which contains five sequence-specific deoxyribonucleases

    NARCIS (Netherlands)

    Reaston, J.; Duybesteyn, M.G.C.; Waard, Adrian de

    1982-01-01

    Five nucleotide sequence-specific deoxyribonucleases present in cell-free extracts of the filamentous cyanobacterium Nostoc PCC7524 have been purified and characterized. One of these enzymes, designated Nsp(7524)I cleaves at a new kind of nucleotide sequence i.e. 5'-PuCATG λ Py-3'. The other four

  3. Secondary metabolite from Nostoc XPORK14A inhibits photosynthesis and growth of Synechocystis PCC 6803.

    Science.gov (United States)

    Shunmugam, Sumathy; Jokela, Jouni; Wahlsten, Matti; Battchikova, Natalia; Ateeq ur Rehman; Vass, Imre; Karonen, Maarit; Sinkkonen, Jari; Permi, Perttu; Sivonen, Kaarina; Aro, Eva-Mari; Allahverdiyeva, Yagut

    2014-06-01

    Screening of 55 different cyanobacterial strains revealed that an extract from Nostoc XPORK14A drastically modifies the amplitude and kinetics of chlorophyll a fluorescence induction of Synechocystis PCC6803 cells.After 2 d exposure to the Nostoc XPORK14A extract, Synechocystis PCC 6803 cells displayed reduced net photosynthetic activity and significantly modified electron transport properties of photosystem II under both light and dark conditions. However, the maximum oxidizable amount of P700 was not strongly affected. The extract also induced strong oxidative stress in Synechocystis PCC 6803 cells in both light and darkness. We identified the secondary metabolite of Nostoc XPORK14A causing these pronounced effects on Synechocystis cells. Mass spectrometry and nuclear magnetic resonance analyses revealed that this compound, designated as M22, has a non-peptide structure. We propose that M22 possesses a dualaction mechanism: firstly, by photogeneration of reactive oxygen species in the presence of light, which in turn affects the photosynthetic machinery of Synechocystis PCC 6803; and secondly, by altering the in vivo redox status of cells, possibly through inhibition of protein kinases.

  4. Taxonomy Icon Data: Synechocystis sp.PCC 6803 [Taxonomy Icon

    Lifescience Database Archive (English)

    Full Text Available Synechocystis sp.PCC 6803 Synechocystis sp.PCC 6803 Synechocystis_sp_PCC_6803_L.png Synecho...cystis_sp_PCC_6803_NL.png Synechocystis_sp_PCC_6803_S.png Synechocystis_sp_PCC_6803_NS.png http://bi...osciencedbc.jp/taxonomy_icon/icon.cgi?i=Synechocystis+sp%2ePCC+6803&t=L http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Synecho...cystis+sp%2ePCC+6803&t=NL http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Synecho...cystis+sp%2ePCC+6803&t=S http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Synechocystis

  5. Viruses Infecting a Freshwater Filamentous Cyanobacterium (Nostoc sp. Encode a Functional CRISPR Array and a Proteobacterial DNA Polymerase B

    Directory of Open Access Journals (Sweden)

    Caroline Chénard

    2016-06-01

    Full Text Available Here we present the first genomic characterization of viruses infecting Nostoc, a genus of ecologically important cyanobacteria that are widespread in freshwater. Cyanophages A-1 and N-1 were isolated in the 1970s and infect Nostoc sp. strain PCC 7210 but remained genomically uncharacterized. Their 68,304- and 64,960-bp genomes are strikingly different from those of other sequenced cyanophages. Many putative genes that code for proteins with known functions are similar to those found in filamentous cyanobacteria, showing a long evolutionary history in their host. Cyanophage N-1 encodes a CRISPR array that is transcribed during infection and is similar to the DR5 family of CRISPRs commonly found in cyanobacteria. The presence of a host-related CRISPR array in a cyanophage suggests that the phage can transfer the CRISPR among related cyanobacteria and thereby provide resistance to infection with competing phages. Both viruses also encode a distinct DNA polymerase B that is closely related to those found in plasmids of Cyanothece sp. strain PCC 7424, Nostoc sp. strain PCC 7120, and Anabaena variabilis ATCC 29413. These polymerases form a distinct evolutionary group that is more closely related to DNA polymerases of proteobacteria than to those of other viruses. This suggests that the polymerase was acquired from a proteobacterium by an ancestral virus and transferred to the cyanobacterial plasmid. Many other open reading frames are similar to a prophage-like element in the genome of Nostoc sp. strain PCC 7524. The Nostoc cyanophages reveal a history of gene transfers between filamentous cyanobacteria and their viruses that have helped to forge the evolutionary trajectory of this previously unrecognized group of phages.

  6. Evolution of space food in Nostoc sp. HK-01

    Science.gov (United States)

    Tomita-Yokotani, Kaori; Yamashita, Masamichi; Hashimoto, Hirofumi; Sato, Seigo; Kimura, Yasuko; Katoh, Hiroshi; Arai, Mayumi

    2012-07-01

    Habitation in outer space is one of our challenges. We have been studying future space agriculture to provide food and oxygen for the habitation area in the space environment, on Mars. A cyanobacteria, Nostoc sp. HK-01, has high several outer space environmental tolerance. We have already confirmed that Nostoc sp.HK-01 had an ability to grow for over several years on the Martian regolith simulant in a laboratory experiment. Nostoc sp HK-01 would have high contribution to change the atmosphere in Mars as a photosynthetic creature. In outer environment, all of materials have to circulate for all of creature living in artificial eco-systems on Mars. This material has several functions as the utilization in space agriculture. Here, we are proposing using them as a food after its growing on Mars. We are trying to determine the best conditions and evolution for space food using Nostoc sp.HK-01 and studying the proposal of utilization of cyanobacteria, Nostoc sp HK-01, for the variation of meal as space agriculture.

  7. Secondary metabolites of cyanobacteria Nostoc sp.

    Science.gov (United States)

    Kobayashi, Akio; Kajiyama, Shin-Ichiro

    1998-03-01

    Cyanobacteria attracted much attention recently because of their secondary metabolites with potent biological activities and unusual structures. This paper reviews some recent studies on the isolation, structural, elucidation and biological activities of the bioactive compounds from cyanobacteria Nostoc species.

  8. Biochemical changes induced by fungicides in nitrogen fixing Nostoc sp.

    Science.gov (United States)

    Deviram, G V N S; Pant, Gaurav; Prasuna, R Gyana

    2013-01-01

    The present study indicates the effect of fungicides (approved by WHO) and their behavior on nitrogen fixer of rice eco system Nostoc sp. Application of plant protecting chemicals at recommended levels braced up the growth of blue green algae thereby enhancing heterocyst formation and nitrogenase activity. Nostoc sp demoed varying degrees of sensitivity to fungicides. Biomass yield, protein, carbohydrate content reduced after 3pg/mL concentration. Heterocyst damage was observed from 4μg/mL, Proline content increased with increase in fungicide concentration, utmost yellowing of the culture started from 4μg/mL. The decreasing order of the toxicity to Nostoc sp with fungicides was Mancozeb> Ediphenphos> Carbendazim> Hexaconazole.

  9. Viruses Infecting a Freshwater Filamentous Cyanobacterium (Nostoc sp.) Encode a Functional CRISPR Array and a Proteobacterial DNA Polymerase B.

    Science.gov (United States)

    Chénard, Caroline; Wirth, Jennifer F; Suttle, Curtis A

    2016-06-14

    Here we present the first genomic characterization of viruses infecting Nostoc, a genus of ecologically important cyanobacteria that are widespread in freshwater. Cyanophages A-1 and N-1 were isolated in the 1970s and infect Nostoc sp. strain PCC 7210 but remained genomically uncharacterized. Their 68,304- and 64,960-bp genomes are strikingly different from those of other sequenced cyanophages. Many putative genes that code for proteins with known functions are similar to those found in filamentous cyanobacteria, showing a long evolutionary history in their host. Cyanophage N-1 encodes a CRISPR array that is transcribed during infection and is similar to the DR5 family of CRISPRs commonly found in cyanobacteria. The presence of a host-related CRISPR array in a cyanophage suggests that the phage can transfer the CRISPR among related cyanobacteria and thereby provide resistance to infection with competing phages. Both viruses also encode a distinct DNA polymerase B that is closely related to those found in plasmids of Cyanothece sp. strain PCC 7424, Nostoc sp. strain PCC 7120, and Anabaena variabilis ATCC 29413. These polymerases form a distinct evolutionary group that is more closely related to DNA polymerases of proteobacteria than to those of other viruses. This suggests that the polymerase was acquired from a proteobacterium by an ancestral virus and transferred to the cyanobacterial plasmid. Many other open reading frames are similar to a prophage-like element in the genome of Nostoc sp. strain PCC 7524. The Nostoc cyanophages reveal a history of gene transfers between filamentous cyanobacteria and their viruses that have helped to forge the evolutionary trajectory of this previously unrecognized group of phages. Filamentous cyanobacteria belonging to the genus Nostoc are widespread and ecologically important in freshwater, yet little is known about the genomic content of their viruses. Here we report the first genomic analysis of cyanophages infecting

  10. Allophycocyanin forms isolated from Nostoc sp. phycobilisomes

    Energy Technology Data Exchange (ETDEWEB)

    Zilinskas, B.A.; Zimmerman, B.K.; Gantt, E.

    1978-01-01

    Allophycocyanin from dissociated phycobilisomes of Nostoc sp. occurs in three spectrally identifiable forms that fractionate on calcium phosphate adsorption chromatography as: allophycocyanin (APC) I (15 to 20%), APC II (40 to 50%), and APC III (30 to 40%). APC I has a single absorption maximum at 654 nm, and a fluorescence emission peak at 678 nm. The absorption peaks of APC II and III are both at 650 nm, but the relative absorbance at 620/650 nm of APC III is less than that of APC II. The emission of both is maximum at 660 nm. On zone sedimentation in sucrose, their S/sub 20 w/ values of 6.0 +- 0.1 (APC I), 5.0 +- 0.1 (APC II), and 5.3 +-0.2 (APC III) were comparable to the order of their elution from Sephadex G-200. On SDS acrylamide gel electrophoresis two subunits were resolved with apparent molecular weights of 16,900 and 18,400 daltons. When stained by Coomassie blue, they were present in a ratio of 1..cap alpha..:1..beta.. in APC II and III, and a probable ratio of 2..cap alpha..:3..beta.. in APC I. The larger size of APC I may be accounted for by additional ..beta.. subunits, by the presence of an additional polypeptide of 35,000 daltons, or both. Over several days, bleaching as noted by a decrease in absorbance at 650 nm, occurred in all three forms; in addition, the more pronounced bleaching at 650 nm, relative to 620 nm, results in APC III becoming spectrally identical to APC II. A trace of a fourth pigment, probably comparable to allophycocyanin-B, was occasionally detected. The results suggest that several in vitro APC forms (sharing similar subunits) arise upon phycobilisome dissociation, and that APC I is the form most closely related to the final fluorescence emitter of intact phycobilisomes. In this form it probably serves as the bridging pigment in energy transfer from the phycobilisomes to chlorophyll.

  11. Multiple ketolases involved in light regulation of canthaxanthin biosynthesis in Nostoc punctiforme PCC 73102.

    Science.gov (United States)

    Schöpf, Lotte; Mautz, Jürgen; Sandmann, Gerhard

    2013-05-01

    In the genome of Nostoc punctiforme PCC 73102, three functional β-carotene ketolase genes exist, one of the crtO and two of the crtW type. They were all expressed and their corresponding enzymes were functional inserting 4-keto groups into β-carotene as shown by functional pathway complementation in Escherichia coli. They all synthesized canthaxanthin but with different efficiencies. Canthaxanthin is the photoprotective carotenoid of N. punctiforme PCC 73102. Under high-light stress, its synthesis was enhanced. This was caused by up-regulation of the transcripts of two genes in combination. The first crtB-encoding phytoene synthase is the gate way enzyme of carotenogenesis resulting in an increased inflow into the pathway. The second was the ketolase gene crtW148 which in high light takes over β-carotene conversion into canthaxanthin from the other ketolases. The other ketolases were down-regulated under high-light conditions. CrtW148 was also exclusively responsible for the last step in 4-keto-myxoxanthophyll synthesis.

  12. Dried Colony in Cyanobacterium, Nostoc sp. HK-01 — Several high Space Environment Tolerances for ``Tanpopo'' Mission

    Science.gov (United States)

    Tomita-Yokotani, K.; Kimura, S.; Kimura, Y.; Igarashi, Y.; Ajioka, R.; Sato, S.; Katoh, H.; Baba, K.

    2013-11-01

    A cyanobacterium, Nostoc sp. HK-01, has high several space environmental tolerance. Nostoc sp HK-01 would have high contribution for the “Tanpopo” mission in Japan Experimental Module of the International Space Station.

  13. Impacts of varying light regimes on phycobiliproteins of Nostoc sp. HKAR-2 and Nostoc sp. HKAR-11 isolated from diverse habitats.

    Science.gov (United States)

    Kannaujiya, Vinod K; Sinha, Rajeshwar P

    2015-11-01

    The adaptability of cyanobacteria in diverse habitats is an important factor to withstand harsh conditions. In the present investigation, the impacts of photosynthetically active radiation (PAR; 400-700 nm), ultraviolet-B (UV-B; 280-315 nm), and PAR + UV-B radiations on two cyanobacteria viz., Nostoc sp. HKAR-2 and Nostoc sp. HKAR-11 inhabiting diverse habitats such as hot springs and rice fields, respectively, were studied. Cell viability was about 14 % in Nostoc sp. HKAR-2 and Nostoc sp. HKAR-11 after 48 h of UV-B exposure. PAR had negligible negative impact on the survival of both cyanobacteria. The continuous exposure of UV-B and PAR + UV-B showed rapid uncoupling, bleaching, fragmentation, and degradation in both phycocyanin (C-PC) and phycoerythrin (C-PE) subunits of phycobiliproteins (PBPs). Remarkable bleaching effect of C-PE and C-PC was not only observed with UV-B or PAR + UV-B radiation, but longer period (24-48 h) of exposure with PAR alone also showed noticeable negative impact. The C-PE and C-PC subunits of the rice field isolate Nostoc sp. HKAR-11 were severely damaged in comparison to the hot spring isolate Nostoc sp. HKAR-2 with rapid wavelength shifting toward shorter wavelengths denoting the bleaching of both the accessory light harvesting pigments. The results indicate that PBPs of the hot spring isolate Nostoc sp. HKAR-2 were more stable under various light regimes in comparison to the rice field isolate Nostoc sp. HKAR-11 that could serve as a good source of valuable pigments to be used in various biomedical and biotechnological applications.

  14. Investigation of a Quadruplex-Forming Repeat Sequence Highly Enriched in Xanthomonas and Nostoc sp.

    Science.gov (United States)

    Rehm, Charlotte; Wurmthaler, Lena A; Li, Yuanhao; Frickey, Tancred; Hartig, Jörg S

    2015-01-01

    In prokaryotes simple sequence repeats (SSRs) with unit sizes of 1-5 nucleotides (nt) are causative for phase and antigenic variation. Although an increased abundance of heptameric repeats was noticed in bacteria, reports about SSRs of 6-9 nt are rare. In particular G-rich repeat sequences with the propensity to fold into G-quadruplex (G4) structures have received little attention. In silico analysis of prokaryotic genomes show putative G4 forming sequences to be abundant. This report focuses on a surprisingly enriched G-rich repeat of the type GGGNATC in Xanthomonas and cyanobacteria such as Nostoc. We studied in detail the genomes of Xanthomonas campestris pv. campestris ATCC 33913 (Xcc), Xanthomonas axonopodis pv. citri str. 306 (Xac), and Nostoc sp. strain PCC7120 (Ana). In all three organisms repeats are spread all over the genome with an over-representation in non-coding regions. Extensive variation of the number of repetitive units was observed with repeat numbers ranging from two up to 26 units. However a clear preference for four units was detected. The strong bias for four units coincides with the requirement of four consecutive G-tracts for G4 formation. Evidence for G4 formation of the consensus repeat sequences was found in biophysical studies utilizing CD spectroscopy. The G-rich repeats are preferably located between aligned open reading frames (ORFs) and are under-represented in coding regions or between divergent ORFs. The G-rich repeats are preferentially located within a distance of 50 bp upstream of an ORF on the anti-sense strand or within 50 bp from the stop codon on the sense strand. Analysis of whole transcriptome sequence data showed that the majority of repeat sequences are transcribed. The genetic loci in the vicinity of repeat regions show increased genomic stability. In conclusion, we introduce and characterize a special class of highly abundant and wide-spread quadruplex-forming repeat sequences in bacteria.

  15. Contribution of two ζ-carotene desaturases to the poly-cis desaturation pathway in the cyanobacterium Nostoc PCC 7120.

    Science.gov (United States)

    Breitenbach, Jürgen; Bruns, Marius; Sandmann, Gerhard

    2013-07-01

    The presence of two completely unrelated ζ-carotene desaturases CrtQa and CrtQb in some Nostoc strains is unique. CrtQb is the ζ-carotene desaturase, which was acquired by almost all cyanobacteria. The additional CrtQa can be regarded as an evolutionary relict of the CrtI desaturase present in non-photosynthetic bacteria. By reconstruction of the carotene desaturation pathway, we showed that both enzymes from Nostoc PCC 7120 were active. However, they differed in their preferred utilization of ζ-carotene Z isomers. CrtQa converted ζ-carotene isomers that were poorly metabolized by CrtQb. In this respect, CrtQa complemented the reactions of CrtQb, which is an advantage avoiding dead ends in the poly-cis desaturation pathway. In addition to ζ-carotene desaturation, CrtQa still possesses the Z to E isomerase function of the ancestral desaturase CrtI. Biochemical characterization showed that CrtQb is an enzyme with one molecule of tightly bound FAD and acts as a dehydrogenase transferring hydrogen to oxidized plastoquinone.

  16. Investigation of a Quadruplex-Forming Repeat Sequence Highly Enriched in Xanthomonas and Nostoc sp.

    Directory of Open Access Journals (Sweden)

    Charlotte Rehm

    Full Text Available In prokaryotes simple sequence repeats (SSRs with unit sizes of 1-5 nucleotides (nt are causative for phase and antigenic variation. Although an increased abundance of heptameric repeats was noticed in bacteria, reports about SSRs of 6-9 nt are rare. In particular G-rich repeat sequences with the propensity to fold into G-quadruplex (G4 structures have received little attention. In silico analysis of prokaryotic genomes show putative G4 forming sequences to be abundant. This report focuses on a surprisingly enriched G-rich repeat of the type GGGNATC in Xanthomonas and cyanobacteria such as Nostoc. We studied in detail the genomes of Xanthomonas campestris pv. campestris ATCC 33913 (Xcc, Xanthomonas axonopodis pv. citri str. 306 (Xac, and Nostoc sp. strain PCC7120 (Ana. In all three organisms repeats are spread all over the genome with an over-representation in non-coding regions. Extensive variation of the number of repetitive units was observed with repeat numbers ranging from two up to 26 units. However a clear preference for four units was detected. The strong bias for four units coincides with the requirement of four consecutive G-tracts for G4 formation. Evidence for G4 formation of the consensus repeat sequences was found in biophysical studies utilizing CD spectroscopy. The G-rich repeats are preferably located between aligned open reading frames (ORFs and are under-represented in coding regions or between divergent ORFs. The G-rich repeats are preferentially located within a distance of 50 bp upstream of an ORF on the anti-sense strand or within 50 bp from the stop codon on the sense strand. Analysis of whole transcriptome sequence data showed that the majority of repeat sequences are transcribed. The genetic loci in the vicinity of repeat regions show increased genomic stability. In conclusion, we introduce and characterize a special class of highly abundant and wide-spread quadruplex-forming repeat sequences in bacteria.

  17. Polyhydroxybutyrate particles in Synechocystis sp PCC 6803: facts and fiction

    Energy Technology Data Exchange (ETDEWEB)

    Tsang, TK; Roberson, RW; Vermaas, WFJ

    2013-09-20

    Transmission electron microscopy has been used to identify poly-3-hydroxybutyrate (PHB) granules in cyanobacteria for over 40 years. Spherical inclusions inside the cell that are electron-transparent and/or slightly electron-dense and that are found in transmission electron micrographs of cyanobacteria are generally assumed to be PHB granules. The aim of this study was to test this assumption in different strains of the cyanobacterium Synechocystis sp. PCC 6803. Inclusions that resemble PHB granules were present in strains lacking a pair of genes essential for PHB synthesis and in wild-type cells under conditions that no PHB granules could be detected by fluorescence staining of PHB. Indeed, in these cells PHB could not be demonstrated chemically by GC/MS either. Based on the results gathered, it is concluded that not all the slightly electron-dense spherical inclusions are PHB granules in Synechocystis sp. PCC 6803. This result is potentially applicable to other cyanobacteria. Alternate assignments for these inclusions are discussed.

  18. Simultaneous Production of Anabaenopeptins and Namalides by the Cyanobacterium Nostoc sp. CENA543.

    Science.gov (United States)

    Shishido, Tânia K; Jokela, Jouni; Fewer, David P; Wahlsten, Matti; Fiore, Marli F; Sivonen, Kaarina

    2017-11-17

    Anabaenopeptins are a diverse group of cyclic peptides, which contain an unusual ureido linkage. Namalides are shorter structural homologues of anabaenopeptins, which also contain an ureido linkage. The biosynthetic origins of namalides are unknown despite a strong resemblance to anabaenopeptins. Here, we show the cyanobacterium Nostoc sp. CENA543 strain producing new (nostamide B-E (2, 4, 5, and 6)) and known variants of anabaenopeptins (schizopeptin 791 (1) and anabaenopeptin 807 (3)). Surprisingly, Nostoc sp. CENA543 also produced namalide B (8) and the new namalides D (7), E (9), and F (10) in similar amounts to anabaenopeptins. Analysis of the complete Nostoc sp. CENA543 genome sequence indicates that both anabaenopeptins and namalides are produced by the same biosynthetic pathway through module skipping during biosynthesis. This unique process involves the skipping of two modules present in different nonribosomal peptide synthetases during the namalide biosynthesis. This skipping is an efficient mechanism since both anabaenopeptins and namalides are synthesized in similar amounts by Nostoc sp. CENA543. Consequently, gene skipping may be used to increase and possibly broaden the chemical diversity of related peptides produced by a single biosynthetic gene cluster. Genome mining demonstrated that the anabaenopeptin gene clusters are widespread in cyanobacteria and can also be found in tectomicrobia bacteria.

  19. The Pkn22 Ser/Thr kinase in Nostoc PCC 7120: role of FurA and NtcA regulators and transcript profiling under nitrogen starvation and oxidative stress.

    Science.gov (United States)

    Yingping, Fan; Lemeille, Sylvain; González, Andrés; Risoul, Véronique; Denis, Yann; Richaud, Pierre; Lamrabet, Otmane; Fillat, Maria F; Zhang, Cheng-Cai; Latifi, Amel

    2015-07-29

    The filamentous cyanobacterium Nostoc sp. strain PCC 7120 can fix N2 when combined nitrogen is not available. Furthermore, it has to cope with reactive oxygen species generated as byproducts of photosynthesis and respiration. We have previously demonstrated the synthesis of Ser/Thr kinase Pkn22 as an important survival response of Nostoc to oxidative damage. In this study we wished to investigate the possible involvement of this kinase in signalling peroxide stress and nitrogen deprivation. Quantitative RT-PCR experiments revealed that the pkn22 gene is induced in response to peroxide stress and to combined nitrogen starvation. Electrophoretic motility assays indicated that the pkn22 promoter is recognized by the global transcriptional regulators FurA and NtcA. Transcriptomic analysis comparing a pkn22-insertion mutant and the wild type strain indicated that this kinase regulates genes involved in important cellular functions such as photosynthesis, carbon metabolism and iron acquisition. Since metabolic changes may lead to oxidative stress, we investigated whether this is the case with nitrogen starvation. Our results rather invalidate this hypothesis thereby suggesting that the function of Pkn22 under nitrogen starvation is independent of its role in response to peroxide stress. Our analyses have permitted a more complete functional description of Ser/Thr kinase in Nostoc. We have decrypted the transcriptional regulation of the pkn22 gene, and analysed the whole set of genes under the control of this kinase in response to the two environmental changes often encountered by cyanobacteria in their natural habitat: oxidative stress and nitrogen deprivation.

  20. Discovery of Rare and Highly Toxic Microcystins from Lichen-Associated Cyanobacterium Nostoc sp. Strain IO-102-I

    OpenAIRE

    Oksanen, Ilona; Jokela, Jouni; Fewer, David P.; Wahlsten, Matti; Rikkinen, Jouko; Sivonen, Kaarina

    2004-01-01

    The production of hepatotoxic cyclic heptapeptides, microcystins, is almost exclusively reported from planktonic cyanobacteria. Here we show that a terrestrial cyanobacterium Nostoc sp. strain IO-102-I isolated from a lichen association produces six different microcystins. Microcystins were identified with liquid chromatography-UV mass spectrometry by their retention times, UV spectra, mass fragmentation, and comparison to microcystins from the aquatic Nostoc sp. strain 152. The dominant micr...

  1. CyanOmics: an integrated database of omics for the model cyanobacterium Synechococcus sp. PCC 7002

    OpenAIRE

    Yang, Yaohua; Feng, Jie; Li, Tao; Ge, Feng; Zhao, Jindong

    2015-01-01

    Cyanobacteria are an important group of organisms that carry out oxygenic photosynthesis and play vital roles in both the carbon and nitrogen cycles of the Earth. The annotated genome of Synechococcus sp. PCC 7002, as an ideal model cyanobacterium, is available. A series of transcriptomic and proteomic studies of Synechococcus sp. PCC 7002 cells grown under different conditions have been reported. However, no database of such integrated omics studies has been constructed. Here we present Cyan...

  2. Nodularin, a cyanobacterial toxin, is synthesized in planta by symbiotic Nostoc sp.

    Science.gov (United States)

    Gehringer, Michelle M; Adler, Lewis; Roberts, Alexandra A; Moffitt, Michelle C; Mihali, Troco K; Mills, Toby J T; Fieker, Claus; Neilan, Brett A

    2012-01-01

    The nitrogen-fixing bacterium, Nostoc, is a commonly occurring cyanobacterium often found in symbiotic associations. We investigated the potential of cycad cyanobacterial endosymbionts to synthesize microcystin/nodularin. Endosymbiont DNA was screened for the aminotransferase domain of the toxin biosynthesis gene clusters. Five endosymbionts carrying the gene were screened for bioactivity. Extracts of two isolates inhibited protein phosphatase 2A and were further analyzed using electrospray ionization mass spectrometry (ESI-MS)/MS. Nostoc sp. ‘Macrozamia riedlei 65.1' and Nostoc sp. ‘Macrozamia serpentina 73.1' both contained nodularin. High performance liquid chromatography (HPLC) HESI-MS/MS analysis confirmed the presence of nodularin at 9.55±2.4 ng μg−1 chlorophyll a in Nostoc sp. ‘Macrozamia riedlei 65.1' and 12.5±8.4 ng μg−1 Chl a in Nostoc sp. ‘Macrozamia serpentina 73.1' extracts. Further scans indicated the presence of the rare isoform [L-Har2] nodularin, which contains ℒ-homoarginine instead of ℒ-arginine. Nodularin was also present at 1.34±0.74 ng ml−1 (approximately 3 pmol per g plant ww) in the methanol root extracts of M. riedlei MZ65, while the presence of [L-Har2] nodularin in the roots of M. serpentina MZ73 was suggested by HPLC HESI-MS/MS analysis. The ndaA-B and ndaF genomic regions were sequenced to confirm the presence of the hybrid polyketide/non-ribosomal gene cluster. A seven amino-acid insertion into the NdaA-C1 domain of N. spumigena NSOR10 protein was observed in all endosymbiont-derived sequences, suggesting the transfer of the nda cluster from N. spumigena to terrestrial Nostoc species. This study demonstrates the synthesis of nodularin and [L-Har2] nodularin in a non-Nodularia species and the production of cyanobacterial hepatotoxin by a symbiont in planta. PMID:22456448

  3. Nodularin, a cyanobacterial toxin, is synthesized in planta by symbiotic Nostoc sp.

    Science.gov (United States)

    Gehringer, Michelle M; Adler, Lewis; Roberts, Alexandra A; Moffitt, Michelle C; Mihali, Troco K; Mills, Toby J T; Fieker, Claus; Neilan, Brett A

    2012-10-01

    The nitrogen-fixing bacterium, Nostoc, is a commonly occurring cyanobacterium often found in symbiotic associations. We investigated the potential of cycad cyanobacterial endosymbionts to synthesize microcystin/nodularin. Endosymbiont DNA was screened for the aminotransferase domain of the toxin biosynthesis gene clusters. Five endosymbionts carrying the gene were screened for bioactivity. Extracts of two isolates inhibited protein phosphatase 2A and were further analyzed using electrospray ionization mass spectrometry (ESI-MS)/MS. Nostoc sp. 'Macrozamia riedlei 65.1' and Nostoc sp. 'Macrozamia serpentina 73.1' both contained nodularin. High performance liquid chromatography (HPLC) HESI-MS/MS analysis confirmed the presence of nodularin at 9.55±2.4 ng μg-1 chlorophyll a in Nostoc sp. 'Macrozamia riedlei 65.1' and 12.5±8.4 ng μg-1 Chl a in Nostoc sp. 'Macrozamia serpentina 73.1' extracts. Further scans indicated the presence of the rare isoform [L-Har(2)] nodularin, which contains L-homoarginine instead of L-arginine. Nodularin was also present at 1.34±0.74 ng ml(-1) (approximately 3 pmol per g plant ww) in the methanol root extracts of M. riedlei MZ65, while the presence of [L-Har(2)] nodularin in the roots of M. serpentina MZ73 was suggested by HPLC HESI-MS/MS analysis. The ndaA-B and ndaF genomic regions were sequenced to confirm the presence of the hybrid polyketide/non-ribosomal gene cluster. A seven amino-acid insertion into the NdaA-C1 domain of N. spumigena NSOR10 protein was observed in all endosymbiont-derived sequences, suggesting the transfer of the nda cluster from N. spumigena to terrestrial Nostoc species. This study demonstrates the synthesis of nodularin and [L-Har(2)] nodularin in a non-Nodularia species and the production of cyanobacterial hepatotoxin by a symbiont in planta.

  4. Flux Balance Analysis of Cyanobacterial Metabolism.The Metabolic Network of Synechocystis sp. PCC 6803

    Czech Academy of Sciences Publication Activity Database

    Knoop, H.; Gründel, M.; Zilliges, Y.; Lehmann, R.; Hoffmann, S.; Lockau, W.; Steuer, Ralf

    2013-01-01

    Roč. 9, č. 6 (2013), e1003081-e1003081 ISSN 1553-7358 R&D Projects: GA MŠk(CZ) EE2.3.20.0256 Institutional support: RVO:67179843 Keywords : SP STRAIN PCC-6803 * SP ATCC 51142 * photoautotrophic metabolism * anacystis-nidulans * reconstructions * pathway * plants * models * growth Subject RIV: EI - Biotechnology ; Bionics Impact factor: 4.829, year: 2013

  5. Phenotypic characterization of Synechocystis sp PCC 6803 substrains reveals differences in sensitivity to abiotic stress

    Czech Academy of Sciences Publication Activity Database

    Zavřel, Tomáš; Očenášová, Petra; Červený, Jan

    2017-01-01

    Roč. 12, č. 12 (2017), č. článku e0189130. E-ISSN 1932-6203 R&D Projects: GA MŠk(CZ) LO1415; GA ČR(CZ) GA15-17367S; GA ČR(CZ) GA15-17367S Institutional support: RVO:86652079 Keywords : sp strain pcc-6803 * high-temperature * sp pcc6803 * cyanobacterium * growth * responses * acclimation * gene * photobioreactor * identification Subject RIV: EI - Biotechnology ; Bionics OBOR OECD: Environmental biotechnology Impact factor: 2.806, year: 2016

  6. Characterization and Optimization of Bioflocculant Exopolysaccharide Production by Cyanobacteria Nostoc sp. BTA97 and Anabaena sp. BTA990 in Culture Conditions.

    Science.gov (United States)

    Tiwari, Onkar Nath; Khangembam, Romi; Shamjetshabam, Minerva; Sharma, Aribam Subhalaxmi; Oinam, Gunapati; Brand, Jerry J

    2015-08-01

    Bioflocculant exopolysaccharide (EPS) production by 40 cyanobacterial strains during their photoautotrophic growth was investigated. Highest levels of EPS were produced by Nostoc sp. BTA97 and Anabaena sp. BTA990. EPS production was maximum during stationary growth phase, when nitrogenase activity was very low. Maximum EPS production occurred at pH 8.0 in the absence of any combined nitrogen source. The cyanobacterial EPS consisted of soluble protein and polysaccharide that included substantial amounts of neutral sugars and uronic acid. The EPS isolated from Anabaena sp. BTA990 and Nostoc sp. BTA97 demonstrated high flocculation capacity. There was a positive correlation between uronic acid content and flocculation activity. The flocculant bound a cationic dye, Alcian Blue, indicating it to be polyanionic. The 16S rRNA gene sequences for Nostoc sp. BTA97 and Anabaena sp. BTA990 were deposited at NCBI GenBank, and accession numbers were obtained as KJ830951 and KJ830948, respectively. The results of these experiments indicate that strains Anabaena sp. BTA990 and Nostoc sp. BTA97 are good candidates for the commercial production of EPS and might be utilized in industrial applications as an alternative to synthetic and abiotic flocculants.

  7. Merocyclophanes C and D from the Cultured Freshwater Cyanobacterium Nostoc sp. (UIC 10110).

    Science.gov (United States)

    May, Daniel S; Chen, Wei-Lun; Lantvit, Daniel D; Zhang, Xiaoli; Krunic, Aleksej; Burdette, Joanna E; Eustaquio, Alessandra; Orjala, Jimmy

    2017-04-28

    Merocyclophanes C and D (1 and 2) were isolated from the cell extract of the cultured cyanobacterium UIC 10110. The structures were determined by one-dimensional nuclear magnetic resonance (NMR) and high-resolution electrospray ionization mass spectrometry and confirmed by 2D NMR techniques. The absolute configurations were determined using electronic circular dichroism spectroscopy. Merocyclophanes C and D represent the first known analogues of the merocyclophane core structure, a recently discovered scaffold of [7,7] paracyclophanes characterized by an α-branched methyl at C-1/C-14; 1 and 2 showed antiproliferative activity against the MDA-MB-435 cell line with IC 50 values of 1.6 and 0.9 μM, respectively. Partial 16S analysis determined UIC 10110 to be a Nostoc sp., and it was found to clade with UIC 10062 Nostoc sp., the only other strain known to produce merocyclophanes. The genome of UIC 10110 was sequenced, and a biosynthetic gene cluster was identified that is proposed to encode type I and type III polyketide synthases that are potentially responsible for production of the merocyclophanes; however, further experiments will be required to verify the true function of the gene cluster. The gene cluster provides a genetic basis for the observed structural differences of the [7,7] paracyclophane core structures.

  8. Effects of Halide Ions on the Carbamidocyclophane Biosynthesis in Nostoc sp. CAVN2

    Science.gov (United States)

    Preisitsch, Michael; Heiden, Stefan E.; Beerbaum, Monika; Niedermeyer, Timo H. J.; Schneefeld, Marie; Herrmann, Jennifer; Kumpfmüller, Jana; Thürmer, Andrea; Neidhardt, Inga; Wiesner, Christoph; Daniel, Rolf; Müller, Rolf; Bange, Franz-Christoph; Schmieder, Peter; Schweder, Thomas; Mundt, Sabine

    2016-01-01

    In this study, the influence of halide ions on [7.7]paracyclophane biosynthesis in the cyanobacterium Nostoc sp. CAVN2 was investigated. In contrast to KI and KF, supplementation of the culture medium with KCl or KBr resulted not only in an increase of growth but also in an up-regulation of carbamidocyclophane production. LC-MS analysis indicated the presence of chlorinated, brominated, but also non-halogenated derivatives. In addition to 22 known cylindrocyclophanes and carbamidocyclophanes, 27 putative congeners have been detected. Nine compounds, carbamidocyclophanes M−U, were isolated, and their structural elucidation by 1D and 2D NMR experiments in combination with HRMS and ECD analysis revealed that they are brominated analogues of chlorinated carbamidocyclophanes. Quantification of the carbamidocyclophanes showed that chloride is the preferably utilized halide, but incorporation is reduced in the presence of bromide. Evaluation of the antibacterial activity of 30 [7.7]paracyclophanes and related derivatives against selected pathogenic Gram-positive and Gram-negative bacteria exhibited remarkable effects especially against methicillin- and vancomycin-resistant staphylococci and Mycobacterium tuberculosis. For deeper insights into the mechanisms of biosynthesis, the carbamidocyclophane biosynthetic gene cluster in Nostoc sp. CAVN2 was studied. The gene putatively coding for the carbamoyltransferase has been identified. Based on bioinformatic analyses, a possible biosynthetic assembly is discussed. PMID:26805858

  9. CyanOmics: an integrated database of omics for the model cyanobacterium Synechococcus sp. PCC 7002.

    Science.gov (United States)

    Yang, Yaohua; Feng, Jie; Li, Tao; Ge, Feng; Zhao, Jindong

    2015-01-01

    Cyanobacteria are an important group of organisms that carry out oxygenic photosynthesis and play vital roles in both the carbon and nitrogen cycles of the Earth. The annotated genome of Synechococcus sp. PCC 7002, as an ideal model cyanobacterium, is available. A series of transcriptomic and proteomic studies of Synechococcus sp. PCC 7002 cells grown under different conditions have been reported. However, no database of such integrated omics studies has been constructed. Here we present CyanOmics, a database based on the results of Synechococcus sp. PCC 7002 omics studies. CyanOmics comprises one genomic dataset, 29 transcriptomic datasets and one proteomic dataset and should prove useful for systematic and comprehensive analysis of all those data. Powerful browsing and searching tools are integrated to help users directly access information of interest with enhanced visualization of the analytical results. Furthermore, Blast is included for sequence-based similarity searching and Cluster 3.0, as well as the R hclust function is provided for cluster analyses, to increase CyanOmics's usefulness. To the best of our knowledge, it is the first integrated omics analysis database for cyanobacteria. This database should further understanding of the transcriptional patterns, and proteomic profiling of Synechococcus sp. PCC 7002 and other cyanobacteria. Additionally, the entire database framework is applicable to any sequenced prokaryotic genome and could be applied to other integrated omics analysis projects. Database URL: http://lag.ihb.ac.cn/cyanomics. © The Author(s) 2015. Published by Oxford University Press.

  10. Hydrogen production by the engineered cyanobacterial strain Nostoc PCC 7120 ΔhupW examined in a flat panel photobioreactor system.

    Science.gov (United States)

    Nyberg, Marcus; Heidorn, Thorsten; Lindblad, Peter

    2015-12-10

    Nitrogenase based hydrogen production was examined in a ΔhupW strain of the filamentous heterocystous cyanobacterium Nostoc PCC 7120, i.e., cells lacking the last step in the maturation system of the large subunit of the uptake hydrogenase and as a consequence with a non-functional uptake hydrogenase. The cells were grown in a developed flat panel photobioreactor system with 3.0L culture volume either aerobically (air) or anaerobically (Ar or 80% N2/20% Ar) and illuminated with a mixture of red and white LED. Aerobic growth of the ΔhupW strain of Nostoc PCC 7120 at 44μmolar photons m(-2)s(-1) PAR gave the highest hydrogen production of 0.7mL H2 L(-1)h(-1), 0.53mmol H2 mg chlorophyll a(-1)h(-1), and a light energy conversion efficiency of 1.2%. Anaerobic growth using 100% argon showed a maximal hydrogen production of 1.7mLL(-1)h(-1), 0.85mmol per mg chlorophyll a(-1) h(-1), and a light energy conversion efficiency of 2.7%. Altering between argon/N2 (20/80) and 100% argon phases resulted in a maximal hydrogen production at hour 128 (100% argon phase) with 6.2mL H2L(-1)h(-1), 0.71mL H2 mg chlorophyll a(-1)h(-1), and a light energy efficiency conversion of 4.0%. The highest buildup of hydrogen gas observed was 6.89% H2 (100% argon phase) of the total photobioreactor system with a maximal production of 4.85mL H2 L(-1)h(-1). The present study clearly demonstrates the potential to use purpose design cyanobacteria in developed flat panel photobioreactor systems for the direct production of the solar fuel hydrogen. Further improvements in the strain used, environmental conditions employed, and growth, production and collection systems used, are needed before a sustainable and economical cyanobacterial based hydrogen production can be realized. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Novel Aeruginosin-865 from Nostoc sp. as a potent anti-inflammatory agent.

    Science.gov (United States)

    Kapuścik, Aleksandra; Hrouzek, Pavel; Kuzma, Marek; Bártová, Simona; Novák, Petr; Jokela, Jouni; Pflüger, Maren; Eger, Andreas; Hundsberger, Harald; Kopecký, Jiří

    2013-11-25

    Aeruginosin-865 (Aer-865), isolated from terrestrial cyanobacterium Nostoc sp. Lukešová 30/93, is the first aeruginosin-type peptide containing both a fatty acid and a carbohydrate moiety, and is the first aeruginosin to be found in the genus Nostoc. Mass spectrometry, chemical and spectroscopic analysis as well as one- and two-dimensional NMR and chiral HPLC analysis of Marfey derivatives were applied to determine the peptidic sequence: D-Hpla, D-Leu, 5-OH-Choi, Agma, with hexanoic and mannopyranosyl uronic acid moieties linked to Choi. We used an AlphaLISA assay to measure the levels of proinflammatory mediators IL-8 and ICAM-1 in hTNF-α-stimulated HLMVECs. Aer-865 showed significant reduction of both: with EC50 values of (3.5±1.5) μg mL(-1) ((4.0±1.7) μM) and (50.0±13.4) μg mL(-1) ((57.8±15.5) μM), respectively. Confocal laser scanning microscopy revealed that the anti-inflammatory effect of Aer-865 was directly associated with inhibition of NF-κB translocation to the nucleus. Moreover, Aer-865 did not show any cytotoxic effect. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Space-environmental tolerances in a cyanobacterium, Nostoc sp. HK-01

    Science.gov (United States)

    Tomita-Yokotani, Kaori; Yokobori, Shin-ichi; Kimura, Shunta; Sato, Seigo; Katoh, Hiroshi; Ajioka, Reiko; Yamagishi, Akihiko; Inoue, Kotomi

    2016-07-01

    We have been investigating the tolerances to space-environments of a cyanobacterium, Nostoc sp. HK-01 (hereafter referred to as HK-01). Dry colonies of HK-01 had high tolerance to dry conditions, but more detailed information about tolerance to high-temperature, UV, gamma-ray and heavy particle beams were not deeply investigated. The obtained dry colonies of HK-01 after exposure to each of the conditions described above were investigated. In all of the tested colonies of HK-01 after exposure, all or some of the cells in the colonies were alive. One of the purposes of space agriculture is growing plants on Mars. In the early stages, of our research, cyanobacteria are introduced on Mars to promote the oxidation of the atmosphere and the formation of soil from Mars's regolith. HK-01 will contribute to each of these factors in the future.

  13. Utilization of a terrestrial cyanobacterium, Nostoc sp. HK-01, for space habitation

    Science.gov (United States)

    Kimura, Shunta; Tomita-Yokotani, Kaori; Arai, Mayumi; Yamashita, Masamichi; Katoh, Hiroshi; Ajioka, Reiko; Inoue, Kotomi

    2016-07-01

    A terrestrial cyanobacterium, Nostoc sp. HK-01 (hereafter HK-01), has several useful abilities for space habitation; photosynthesis, nitrogen fixation, and space environmental tolerances to vacuum, UV, gamma-ray, heavy particle beam, low and high temperature. Space environmental tolerances are important for transportation to Mars. HK-01 can grow on Martian regolith simulant (MRS) in vitro. Furthermore, HK-01 is useful as food. HK-01 may be utilized as oxygen supply, soil formation and food material for bio-chemical circulation in closed bio-ecosystems, including space habitation such as Mars. HK-01 was adopted as a biological material for the "TANPOPO" mission (JAXA et al.,), because of their high environmental tolerances. The "TANPOPO" mission is performing the space exposure experiments on the Japan Experimental Module (JEM) of the International Space Station (ISS). The results of these experiments will show the ability of HK-01 to survive in space.

  14. Physicochemical parameters optimization, and purification of phycobiliproteins from the isolated Nostoc sp.

    Science.gov (United States)

    Johnson, Eldin M; Kumar, Kanhaiya; Das, Debabrata

    2014-08-01

    The present study investigated the effects of several physicochemical parameters on the improvement of phycobiliproteins (especially phycocyanin) synthesis in a newly isolated species of Nostoc sp. Standard BG11₀ medium was modified to enhance the biomass productivity in different photobioreactors. The initial pH of 8, light intensity of 40 μmol m(-2)s(-1), temperature of 35 °C, diurnal cycle of 16:8 h (light:dark regime), 75.48 μM Na₂CO₃ and 17.65 mM NaNO₃ were found most suitable for the phycobiliproteins synthesis. Cyanobacteria exhibited chromatic adaptation, causing overexpression of phycocyanin in red and phycoerythrin in green light. The maximum phycobiliproteins yield of 0.13 gg(-1) dry cell weight was obtained in green light. Phycocyanin was further purified using thin layer chromatography (TLC), anion exchange chromatography and SDS-PAGE (denaturing gel) electrophoresis. Copyright © 2014 Elsevier Ltd. All rights reserved.

  15. Production of High Amounts of Hepatotoxin Nodularin and New Protease Inhibitors Pseudospumigins by the Brazilian Benthic Nostoc sp. CENA543

    Science.gov (United States)

    Jokela, Jouni; Heinilä, Lassi M. P.; Shishido, Tânia K.; Wahlsten, Matti; Fewer, David P.; Fiore, Marli F.; Wang, Hao; Haapaniemi, Esa; Permi, Perttu; Sivonen, Kaarina

    2017-01-01

    Nostoc is a cyanobacterial genus, common in soils and a prolific producer of natural products. This research project aimed to explore and characterize Brazilian cyanobacteria for new bioactive compounds. Here we report the production of hepatotoxins and new protease inhibitors from benthic Nostoc sp. CENA543 isolated from a small, shallow, saline-alkaline lake in the Nhecolândia, Pantanal wetland area in Brazil. Nostoc sp. CENA543 produces exceptionally high amounts of nodularin-R. This is the first free-living Nostoc that produces nodularin at comparable levels as the toxic, bloom-forming, Nodularia spumigena. We also characterized pseudospumigins A–F, which are a novel family of linear tetrapeptides. Pseudospumigins are structurally related to linear tetrapeptide spumigins and aeruginosins both present in N. spumigena but differ in respect to their diagnostic amino acid, which is Ile/Leu/Val in pseudospumigins, Pro/mPro in spumigins, and Choi in aeruginosins. The pseudospumigin gene cluster is more similar to the spumigin biosynthetic gene cluster than the aeruginosin gene cluster. Pseudospumigin A inhibited trypsin (IC50 4.5 μM after 1 h) in a similar manner as spumigin E from N. spumigena but was almost two orders of magnitude less potent. This study identifies another location and environment where the hepatotoxic nodularin has the potential to cause the death of eukaryotic organisms. PMID:29062311

  16. Production of High Amounts of Hepatotoxin Nodularin and New Protease Inhibitors Pseudospumigins by the Brazilian Benthic Nostoc sp. CENA543

    Directory of Open Access Journals (Sweden)

    Jouni Jokela

    2017-10-01

    Full Text Available Nostoc is a cyanobacterial genus, common in soils and a prolific producer of natural products. This research project aimed to explore and characterize Brazilian cyanobacteria for new bioactive compounds. Here we report the production of hepatotoxins and new protease inhibitors from benthic Nostoc sp. CENA543 isolated from a small, shallow, saline-alkaline lake in the Nhecolândia, Pantanal wetland area in Brazil. Nostoc sp. CENA543 produces exceptionally high amounts of nodularin-R. This is the first free-living Nostoc that produces nodularin at comparable levels as the toxic, bloom-forming, Nodularia spumigena. We also characterized pseudospumigins A–F, which are a novel family of linear tetrapeptides. Pseudospumigins are structurally related to linear tetrapeptide spumigins and aeruginosins both present in N. spumigena but differ in respect to their diagnostic amino acid, which is Ile/Leu/Val in pseudospumigins, Pro/mPro in spumigins, and Choi in aeruginosins. The pseudospumigin gene cluster is more similar to the spumigin biosynthetic gene cluster than the aeruginosin gene cluster. Pseudospumigin A inhibited trypsin (IC50 4.5 μM after 1 h in a similar manner as spumigin E from N. spumigena but was almost two orders of magnitude less potent. This study identifies another location and environment where the hepatotoxic nodularin has the potential to cause the death of eukaryotic organisms.

  17. Role of Two Cell Wall Amidases in Septal Junction and Nanopore Formation in the Multicellular Cyanobacterium Anabaena sp. PCC 7120

    Directory of Open Access Journals (Sweden)

    Jan Bornikoel

    2017-09-01

    Full Text Available Filamentous cyanobacteria have developed a strategy to perform incompatible processes in one filament by differentiating specialized cell types, N2-fixing heterocysts and CO2-fixing, photosynthetic, vegetative cells. These bacteria can be considered true multicellular organisms with cells exchanging metabolites and signaling molecules via septal junctions, involving the SepJ and FraCD proteins. Previously, it was shown that the cell wall lytic N-acetylmuramyl-L-alanine amidase, AmiC2, is essential for cell–cell communication in Nostoc punctiforme. This enzyme perforates the septal peptidoglycan creating an array of nanopores, which may be the framework for septal junction complexes. In Anabaena sp. PCC 7120, two homologs of AmiC2, encoded by amiC1 and amiC2, were identified and investigated in two different studies. Here, we compare the function of both AmiC proteins by characterizing different Anabaena amiC mutants, which was not possible in N. punctiforme, because there the amiC1 gene could not be inactivated. This study shows the different impact of each protein on nanopore array formation, the process of cell–cell communication, septal protein localization, and heterocyst differentiation. Inactivation of either amidase resulted in significant reduction in nanopore count and in the rate of fluorescent tracer exchange between neighboring cells measured by FRAP analysis. In an amiC1 amiC2 double mutant, filament morphology was affected and heterocyst differentiation was abolished. Furthermore, the inactivation of amiC1 influenced SepJ localization and prevented the filament-fragmentation phenotype that is characteristic of sepJ or fraC fraD mutants. Our findings suggest that both amidases are to some extent redundant in their function, and describe a functional relationship of AmiC1 and septal proteins SepJ and FraCD.

  18. Phylogenetically distant clade of Nostoc-like taxa with the description of Aliinostoc gen. nov. and Aliinostoc morphoplasticum sp. nov.

    Science.gov (United States)

    Bagchi, Suvendra Nath; Dubey, Neelam; Singh, Prashant

    2017-09-01

    Nostoc is a complex and tough genus to differentiate, and its morphological plasticity makes it taxonomically complicated. Its cryptic diversity and almost no distinguishable morphological characteristics make this genus incredibly heterogeneous to evaluate on taxonomic scales. The strain NOS, isolated from a eutrophic water body, is being described as a new genus Aliinostoc with the strain showing motile hormogonia with gas vesicles as an atypical feature, which is currently considered as the diacritical feature of the genus but should be subjected to critical evaluation in the near future. The phylogenetic placement of Aliinostoc along with some other related sequences of Nostoc clearly separated this clade from Nostoc sensu stricto with high bootstrap support and robust topology in all the methods tested, thus providing strong proof of the taxa being representative of a new genus which morphologically appears to be Nostoc-like. Subsequent phylogenetic assessment using the rbcL, psbA, rpoC1 and tufA genes was done with the aim of facilitating future multi-locus studies on the proposed genus for better taxonomic clarity and resolution. Folding of the 16S-23S internal transcribed spacer region and subsequent comparisons with members of the genera Nostoc, Anabaena, Aulosira, Cylindrospermum, Sphaerospermopsis, Raphidiopsis, Desmonostoc and Mojavia gave entirely new secondary structures for the D1-D1' and box-B helix. Clear and separate clustering from Nostoc sensu stricto supports the establishment of Aliinostoc gen. nov. with the type species being Aliinostoc morphoplasticum sp. nov. in accordance with the International Code of Nomenclature for algae, fungi and plants.

  19. Deep sequencing-based identification of small regulatory RNAs in Synechocystis sp. PCC 6803.

    Directory of Open Access Journals (Sweden)

    Wen Xu

    Full Text Available Synechocystis sp. PCC 6803 is a genetically tractable model organism for photosynthesis research. The genome of Synechocystis sp. PCC 6803 consists of a circular chromosome and seven plasmids. The importance of small regulatory RNAs (sRNAs as mediators of a number of cellular processes in bacteria has begun to be recognized. However, little is known regarding sRNAs in Synechocystis sp. PCC 6803. To provide a comprehensive overview of sRNAs in this model organism, the sRNAs of Synechocystis sp. PCC 6803 were analyzed using deep sequencing, and 7,951,189 reads were obtained. High quality mapping reads (6,127,890 were mapped onto the genome and assembled into 16,192 transcribed regions (clusters based on read overlap. A total number of 5211 putative sRNAs were revealed from the genome and the 4 megaplasmids, and 27 of these molecules, including four from plasmids, were confirmed by RT-PCR. In addition, possible target genes regulated by all of the putative sRNAs identified in this study were predicted by IntaRNA and analyzed for functional categorization and biological pathways, which provided evidence that sRNAs are indeed involved in many different metabolic pathways, including basic metabolic pathways, such as glycolysis/gluconeogenesis, the citrate cycle, fatty acid metabolism and adaptations to environmentally stress-induced changes. The information from this study provides a valuable reservoir for understanding the sRNA-mediated regulation of the complex physiology and metabolic processes of cyanobacteria.

  20. Production of squalene in Synechocystis sp. PCC 6803.

    Directory of Open Access Journals (Sweden)

    Elias Englund

    Full Text Available In recent years, there has been an increased interest in the research and development of sustainable alternatives to fossil fuels. Using photosynthetic microorganisms to produce such alternatives is advantageous, since they can achieve direct conversion of carbon dioxide from the atmosphere into the desired product, using sunlight as the energy source. Squalene is a naturally occurring 30-carbon isoprenoid, which has commercial use in cosmetics and in vaccines. If it could be produced sustainably on a large scale, it could also be used instead of petroleum as a raw material for fuels and as feedstock for the chemical industry. The unicellular cyanobacterium Synechocystis PCC 6803 possesses a gene, slr2089, predicted to encode squalene hopene cyclase (Shc, an enzyme converting squalene into hopene, the substrate for forming hopanoids. Through inactivation of slr2089 (shc, we explored the possibility to produce squalene using cyanobacteria. The inactivation led to accumulation of squalene, to a level over 70 times higher than in wild type cells, reaching 0.67 mg OD750(-1 L(-1. We did not observe any significant growth deficiency in the Δshc strain compared to the wild type Synechocystis, even at high light conditions, suggesting that the observed squalene accumulation was not detrimental to growth, and that formation of hopene by Shc is not crucial for growth under normal conditions, nor for high-light stress tolerance. Effects of different light intensities and growth stages on squalene accumulation in the Δshc strain were investigated. We also identified a gene, sll0513, as a putative squalene synthase in Synechocystis, and verified its function by inactivation. In this work, we show that it is possible to use the cyanobacterium Synechocystis to generate squalene, a hydrocarbon of commercial interest and a potential biofuel. We also report the first identification of a squalene hopene cyclase, and the second identification of squalene synthase

  1. Complete genome sequence of cyanobacterium Nostoc sp. NIES-3756, a potentially useful strain for phytochrome-based bioengineering.

    Science.gov (United States)

    Hirose, Yuu; Fujisawa, Takatomo; Ohtsubo, Yoshiyuki; Katayama, Mitsunori; Misawa, Naomi; Wakazuki, Sachiko; Shimura, Yohei; Nakamura, Yasukazu; Kawachi, Masanobu; Yoshikawa, Hirofumi; Eki, Toshihiko; Kanesaki, Yu

    2016-01-20

    To explore the diverse photoreceptors of cyanobacteria, we isolated Nostoc sp. strain NIES-3756 from soil at Mimomi-Park, Chiba, Japan, and determined its complete genome sequence. The Genome consists of one chromosome and two plasmids (total 6,987,571 bp containing no gaps). The NIES-3756 strain carries 7 phytochrome and 12 cyanobacteriochrome genes, which will facilitate the studies of phytochrome-based bioengineering. Copyright © 2015. Published by Elsevier B.V.

  2. Biochemical characterization of putative adenylate dimethylallyltransferase and cytokinin dehydrogenase from Nostoc sp. PCC 7120

    NARCIS (Netherlands)

    Frébortová, Jitka; Greplová, Marta; Seidl, Michael F.; Heyl, Alexander; Frébort, Ivo

    2015-01-01

    Cytokinins, a class of phytohormones, are adenine derivatives common to many different organisms. In plants, these play a crucial role as regulators of plant development and the reaction to abiotic and biotic stress. Key enzymes in the cytokinin synthesis and degradation in modern land plants are

  3. Molecular exploration of the highly radiation resistant cyanobacterium Arthrospira sp. PCC 8005

    Science.gov (United States)

    Badri, Hanène; Leys, Natalie; Wattiez, Ruddy

    Arthrospira (Spirulina) is a photosynthetic cyanobacterium able to use sunlight to release oxygen from water and remove carbon dioxide and nitrate from water. In addition, it is suited for human consumption (edible). For these traits, the cyanobacterium Arthrospira sp. PCC 8005 was selected by the European Space Agency (ESA) as part of the life support system MELiSSA for recycling oxygen, water, and food during future long-haul space missions. However, during such extended missions, Arthrospira sp. PCC 8005 will be exposed to continuous artificial illumination and harmful cosmic radiation. The aim of this study was to investigate how Arthrospira will react and behave when exposed to such stress environment. The cyanobacterium Arthrospira sp. PCC 8005 was exposed to high gamma rays doses in order to unravel in details the response of this bacterium following such stress. Test results showed that after acute exposure to high doses of 60Co gamma radiation upto 3200 Gy, Arthrospira filaments were still able to restart photosynthesis and proliferate normally. Doses above 3200 Gy, did have a detrimental effect on the cells, and delayed post-irradiation proliferation. The photosystem activity, measured as the PSII quantum yield immediately after irradiation, decreased significantly at radiation doses above 3200 Gy. Likewise through pigment content analysis a significant decrease in phycocyanin was observed following exposure to 3200 Gy. The high tolerance of this bacterium to 60Co gamma rays (i.e. ca. 1000x more resistant than human cells for example) raised our interest to investigate in details the cellular and molecular mechanisms behind this amazing resistance. Optimised DNA, RNA and protein extraction methods and a new microarray chip specific for Arthrospira sp. PCC 8005 were developed to identify the global cellular and molecular response following exposure to 3200 Gy and 5000 Gy A total of 15,29 % and 30,18 % genes were found differentially expressed in RNA

  4. Glycogen production for biofuels by the euryhaline cyanobacteria Synechococcus sp. strain PCC 7002 from an oceanic environment.

    Science.gov (United States)

    Aikawa, Shimpei; Nishida, Atsumi; Ho, Shih-Hsin; Chang, Jo-Shu; Hasunuma, Tomohisa; Kondo, Akihiko

    2014-01-01

    Oxygenic photosynthetic microorganisms such as cyanobacteria and microalgae have attracted attention as an alternative carbon source for the next generation of biofuels. Glycogen abundantly accumulated in cyanobacteria is a promising feedstock which can be converted to ethanol through saccharification and fermentation processes. In addition, the utilization of marine cyanobacteria as a glycogen producer can eliminate the need for a freshwater supply. Synechococcus sp. strain PCC 7002 is a fast-growing marine coastal euryhaline cyanobacteria, however, the glycogen yield has not yet been determined. In the present study, the effects of light intensity, CO2 concentration, and salinity on the cell growth and glycogen content were investigated in order to maximize glycogen production in Synechococcus sp. strain PCC 7002. The optimal culture conditions for glycogen production in Synechococcus sp. strain PCC 7002 were investigated. The maximum glycogen production of 3.5 g L(-1) for 7 days (a glycogen productivity of 0.5 g L(-1) d(-1)) was obtained under a high light intensity, a high CO2 level, and a nitrogen-depleted condition in brackish water. The glycogen production performance in Synechococcus sp. strain PCC 7002 was the best ever reported in the α-polyglucan (glycogen or starch) production of cyanobacteria and microalgae. In addition, the robustness of glycogen production in Synechococcus sp. strain PCC 7002 to salinity was evaluated in seawater and freshwater. The peak of glycogen production of Synechococcus sp. strain PCC 7002 in seawater and freshwater were 3.0 and 1.8 g L(-1) in 7 days, respectively. Glycogen production in Synechococcus sp. strain PCC 7002 maintained the same level in seawater and half of the level in freshwater compared with the optimal result obtained in brackish water. We conclude that Synechococcus sp. strain PCC 7002 has high glycogen production activity and glycogen can be provided from coastal water accompanied by a fluctuation

  5. Effects of fatty acid activation on photosynthetic production of fatty acid-based biofuels in Synechocystis sp. PCC6803

    Directory of Open Access Journals (Sweden)

    Gao Qianqian

    2012-03-01

    Full Text Available Abstract Background Direct conversion of solar energy and carbon dioxide to drop in fuel molecules in a single biological system can be achieved from fatty acid-based biofuels such as fatty alcohols and alkanes. These molecules have similar properties to fossil fuels but can be produced by photosynthetic cyanobacteria. Results Synechocystis sp. PCC6803 mutant strains containing either overexpression or deletion of the slr1609 gene, which encodes an acyl-ACP synthetase (AAS, have been constructed. The complete segregation and deletion in all mutant strains was confirmed by PCR analysis. Blocking fatty acid activation by deleting slr1609 gene in wild-type Synechocystis sp. PCC6803 led to a doubling of the amount of free fatty acids and a decrease of alkane production by up to 90 percent. Overexpression of slr1609 gene in the wild-type Synechocystis sp. PCC6803 had no effect on the production of either free fatty acids or alkanes. Overexpression or deletion of slr1609 gene in the Synechocystis sp. PCC6803 mutant strain with the capability of making fatty alcohols by genetically introducing fatty acyl-CoA reductase respectively enhanced or reduced fatty alcohol production by 60 percent. Conclusions Fatty acid activation functionalized by the slr1609 gene is metabolically crucial for biosynthesis of fatty acid derivatives in Synechocystis sp. PCC6803. It is necessary but not sufficient for efficient production of alkanes. Fatty alcohol production can be significantly improved by the overexpression of slr1609 gene.

  6. Anti-MRSA-acting carbamidocyclophanes H-L from the Vietnamese cyanobacterium Nostoc sp. CAVN2.

    Science.gov (United States)

    Preisitsch, Michael; Harmrolfs, Kirsten; Pham, Hang T L; Heiden, Stefan E; Füssel, Anna; Wiesner, Christoph; Pretsch, Alexander; Swiatecka-Hagenbruch, Monika; Niedermeyer, Timo H J; Müller, Rolf; Mundt, Sabine

    2015-03-01

    The methanol extract of the Vietnamese freshwater cyanobacterium Nostoc sp. CAVN2 exhibited cytotoxic effects against MCF-7 and 5637 cancer cell lines as well as against nontumorigenic FL and HaCaT cells and was active against methicillin-resistant Staphylococcus aureus (MRSA) and Streptococcus pneumoniae. High-resolution mass spectrometric analysis indicated the presence of over 60 putative cyclophane-like compounds in an antimicrobially active methanol extract fraction. A paracyclophanes-focusing extraction and separation methodology led to the isolation of 5 new carbamidocyclophanes (1-5) and 11 known paracyclophanes (6-16). The structures and their stereochemical configurations were elucidated by a combination of spectrometric and spectroscopic methods including HRMS, 1D and 2D NMR analyses and detailed comparative CD analysis. The newly described monocarbamoylated [7.7]paracyclophanes (1, 2, 4 and 5) differ by a varying degree of chlorination in the side chains. Carbamidocyclophane J (3) is the very first reported carbamidocyclophane bearing a single halogenation in both butyl residues. Based on previous studies a detailed phylogenetic examination of cyclophane-producing cyanobacteria was carried out. The biological evaluation of 1-16 against various clinical pathogens highlighted a remarkable antimicrobial activity against MRSA with MICs of 0.1-1.0 μM, and indicated that the level of antibacterial activity is related to the presence of carbamoyl moieties.

  7. Raman spectroscopic analysis of the responds of desert cyanobacterium Nostoc sp under UV-B radiation

    Science.gov (United States)

    Wang, Gaohong; Hao, Zongjie; Hu, Chunxiang; Liu, Yongding

    Cyanobacteria are renowned for tolerating extremes of desiccation, UV radiation, freezethaw cycles, hypersalinity and oligotrophy, which make them as candidate par excellence for terraforming in extraterrestrial planet. Recently Raman spectrum was applied to study the biochemical information changes in different field of life science. In this study, we investigated the respond of desert cyanobactreium Nostoc sp under UV-B radiation via FT-Raman spectra. It was found that the spectral biomarkers of protectant molecular of UV radiation such as β-carotene and scytonemin were induced by UV-B radiation, but Chlorophyll a content was decreased, and also the photosynthesis activity was inhibited significantly. After light adaptation without UV-B radiation, the Chlorophyll a content and photosynthesis activity returned to high level, butβ-carotene and scytonemin content remained in the cells. Those results indicated that desert Cyanobacteria have good adaptation ability for UV-B radiation and synthesis of protectant molecular may be an effective strategy for its adaptation in evolution.

  8. Structural elucidation and molecular docking of a novel antibiotic compound from cyanobacterium Nostoc sp. MGL001

    Directory of Open Access Journals (Sweden)

    Niveshika No Name

    2016-11-01

    Full Text Available Cyanobacteria are rich source of array of bioactive compounds. The present study reports a novel antibacterial bioactive compound purified from cyanobacterium Nostoc sp. MGL001 using various chromatographic techniques viz. thin layer chromatography (TLC and high performance liquid chromatography (HPLC. Further characterization was done using electrospray ionisation mass spectroscopy (ESIMS and nuclear magnetic resonance (NMR and predicted structure of bioactive compound was 9-Ethyliminomethyl-12-(morpholin - 4 - ylmethoxy -5, 8, 13, 16 – tetraaza – hexacene - 2, 3 dicarboxylic acid (EMTAHDCA. Structure of EMTAHDCA clearly indicated that it is a novel compound that was not reported in literature or natural product database. The compound exhibited growth inhibiting effects mainly against the gram negative bacterial strains and produced maximum zone of inhibition at 150 μg/mL concentration. The compound was evaluated through in silico studies for its ability to bind 30S ribosomal fragment (PDB ID: 1YRJ, 1MWL, 1J7T and 1LC4 and OmpF porin protein (4GCP, 4GCQ and 4GCS which are the common targets of various antibiotic drugs. Comparative molecular docking study revealed that EMTAHDCA has strong binding affinity for these selected targets in comparison to a number of most commonly used antibiotics. The ability of EMTAHDCA to bind the active sites on the proteins and 30S ribosomal fragments where the antibiotic drugs generally bind indicated that it is functionally similar to the commercially available drugs.

  9. Coping with iron limitation: a metabolomic study of Synechocystis sp PCC 6803

    Czech Academy of Sciences Publication Activity Database

    Rivas-Ubach, A.; Poret-Peterson, A. T.; Penuelas, J.; Sardans, J.; Pérez-Trujillo, M.; Legido-Quigley, C.; Oravec, Michal; Urban, Otmar; Elser, J. J.

    2018-01-01

    Roč. 40, č. 2 (2018), č. článku 28. ISSN 0137-5881 R&D Projects: GA MŠk(CZ) LO1415; GA MŠk(CZ) LM2015061 Institutional support: RVO:86652079 Keywords : sp strain pcc-6803 * ocean acidification * unicellular cyanobacterium * marine-phytoplankton * foliar metabolomes * nitrogen-fixation * metal homeostasis * oxidative stress * pacific-ocean * responses * Metabolomics * Metallomics * Iron limitation * Cyanobacteria * Ecological stoichiometry Subject RIV: EH - Ecology, Behaviour OBOR OECD: Environmental sciences (social aspects to be 5.7) Impact factor: 1.364, year: 2016

  10. ORF Sequence: NC_003240 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available Nostoc sp. PCC 7120] MEYPQINIKLLSEEDRELVKQAKQRAEDLRLTFKEFVLDCIRNALFEESPDDADSATAAEIEALKAQIAALSEKVNIPSASKTEVHTLQERLNQLRVGISNEIKKDREQLASLALALSRLESQIEQLVPSLNLQVDGEVNSNSDGDWLFGEDSGAELPLT

  11. Protein (Cyanobacteria): 427708671 [PGDBj - Ortholog DB

    Lifescience Database Archive (English)

    Full Text Available H endonuclease Nostoc sp. PCC 7107 MSSLYINAELRRLVARRADYICEYCLVSESDRSSGCQVDHIISVKHGGATTADNLCYACIFCNLQKGTDLGSINWQTGELVRFFNPRRDFWGEHFRLGEGVIQPLTDIGEVTARIFDFNCDERVIERQALILSGQYPSKSALKRINK

  12. Physiological and biochemical responses of Synechococcus sp. PCC7942 to Irgarol 1051 and diuron.

    Science.gov (United States)

    Deng, Xiangyuan; Gao, Kun; Sun, Junlong

    2012-10-15

    Cyanobacteria are prokaryotic algae found in oceans and freshwaters worldwide. These organisms are important primary producers in aquatic ecosystems because they can provide essential food for grazers and herbivores. In this study, the physiological and biochemical responses of the freshwater cyanobacterium Synechococcus sp. PCC7942 to two organic booster biocides Irgarol 1051 and diuron were compared and evaluated using 96 h growth tests in a batch-culture system. The 96 h median effective concentrations (EC(50)) were 0.019 and 0.097 μmol L(-1) for Irgarol 1051 and diuron, respectively, which indicate that Irgarol 1051 is about 5 times more toxic than diuron to cyanobacteria. Moreover, remarkable physiological and biochemical responses occurred in the Irgarol 1051 and diuron treatments. Irgarol 1051 and diuron stimulated cyanobacterial growth, increased the soluble protein content, and enhanced the catalase (CAT) activity at low concentrations, but inhibited them at high concentrations. However, the malondialdehyde (MDA) and polysaccharide content of the cyanobacteria were only significantly affected by Irgarol 1051. These observations suggest that Irgarol 1051 and diuron are toxic to Synechococcus sp. PCC7942, and their use should be restricted in maritime industries. Copyright © 2012 Elsevier B.V. All rights reserved.

  13. In silico characterization and transcriptomic analysis of nif family genes from Anabaena sp. PCC7120.

    Science.gov (United States)

    Singh, Shilpi; Shrivastava, Alok Kumar

    2017-10-01

    In silico approaches in conjunction with morphology, nitrogenase activity, and qRT-PCR explore the impact of selected abiotic stressor such as arsenic, salt, cadmium, copper, and butachlor on nitrogen fixing (nif family) genes of diazotrophic cyanobacterium Anabaena sp. PCC7120. A total of 19 nif genes are present within the Anabaena genome that is involved in the process of nitrogen fixation. Docking studies revealed the interaction between these nif gene-encoded proteins and the selected abiotic stressors which were further validated through decreased heterocyst frequency, fragmentation of filaments, and downregulation of nitrogenase activity under these stresses indicating towards their toxic impact on nitrogen fixation potential of filamentous cyanobacterium Anabaena sp. PCC7120. Another appealing finding of this study is even though having similar binding energy and similar interacting residues between arsenic/salt and copper/cadmium to nif-encoded proteins, arsenic and cadmium are more toxic than salt and copper for nitrogenase activity of Anabaena which is crucial for growth and yield of rice paddy and soil reclamation.

  14. The Anabaena sp. PCC 7120 Exoproteome: Taking a Peek outside the Box

    Directory of Open Access Journals (Sweden)

    Paulo Oliveira

    2015-01-01

    Full Text Available The interest in examining the subset of proteins present in the extracellular milieu, the exoproteome, has been growing due to novel insights highlighting their role on extracellular matrix organization and biofilm formation, but also on homeostasis and development. The cyanobacterial exoproteome is poorly studied, and the role of cyanobacterial exoproteins on cell wall biogenesis, morphology and even physiology is largely unknown. Here, we present a comprehensive examination of the Anabaena sp. PCC 7120 exoproteome under various growth conditions. Altogether, 139 proteins belonging to 16 different functional categories have been identified. A large fraction (48% of the identified proteins is classified as “hypothetical”, falls into the “other categories” set or presents no similarity to other proteins. The evidence presented here shows that Anabaena sp. PCC 7120 is capable of outer membrane vesicle formation and that these vesicles are likely to contribute to the exoproteome profile. Furthermore, the activity of selected exoproteins associated with oxidative stress has been assessed, suggesting their involvement in redox homeostasis mechanisms in the extracellular space. Finally, we discuss our results in light of other cyanobacterial exoproteome studies and focus on the potential of exploring cyanobacteria as cell factories to produce and secrete selected proteins.

  15. Root colonization and phytostimulation by phytohormones producing entophytic Nostoc sp. AH-12.

    Science.gov (United States)

    Hussain, Anwar; Hamayun, Muhammad; Shah, Syed Tariq

    2013-11-01

    Nostoc, a nitrogen-fixing cyanobacterium, has great potential to make symbiotic associations with a wide range of plants and benefit its hosts with nitrogen in the form of nitrates. It may also use phytohormones as a tool to promote plant growth. Phytohormones [cytokinin (Ck) and IAA] were determined in the culture of an endophytic Nostoc isolated from rice roots. The strain was able to accumulate as well as release phytohormones to the culture media. Optimum growth conditions for the production of zeatin and IAA were a temperature of 25 °C and a pH of 8.0. Time-dependent increase in the accumulation and release of phytohormones was recorded. To evaluate the impact of cytokinins, an ipt knockout mutant in the background of Nostoc was generated by homologous recombination method. A sharp decline (up to 80 %) in the zeatin content was observed in the culture of mutant strain Nostoc AHM-12. Association of the mutant and wild type strain with rice and wheat roots was studied under axenic conditions. The efficacy of Nostoc to colonize plant root was significantly reduced (P Nostoc to colonize plant root and promote its growth.

  16. The use of fluorescence microscopy and image analysis for rapid detection of non-producing revertant cells of Synechocystis sp. PCC6803 and Synechococcus sp. PCC7002.

    Science.gov (United States)

    Schulze, Katja; Lang, Imke; Enke, Heike; Grohme, Diana; Frohme, Marcus

    2015-04-17

    Ethanol production via genetically engineered cyanobacteria is a promising solution for the production of biofuels. Through the introduction of a pyruvate decarboxylase and alcohol dehydrogenase direct ethanol production becomes possible within the cells. However, during cultivation genetic instability can lead to mutations and thus loss of ethanol production. Cells then revert back to the wild type phenotype. A method for a rapid and simple detection of these non-producing revertant cells in an ethanol producing cell population is an important quality control measure in order to predict genetic stability and the longevity of a producing culture. Several comparable cultivation experiments revealed a difference in the pigmentation for non-producing and producing cells: the accessory pigment phycocyanin (PC) is reduced in case of the ethanol producer, resulting in a yellowish appearance of the culture. Microarray and western blot studies of Synechocystis sp. PCC6803 and Synechococcus sp. PCC7002 confirmed this PC reduction on the level of RNA and protein. Based on these findings we developed a method for fluorescence microscopy in order to distinguish producing and non-producing cells with respect to their pigmentation phenotype. By applying a specific filter set the emitted fluorescence of a producer cell with a reduced PC content appeared orange. The emitted fluorescence of a non-producing cell with a wt pigmentation phenotype was detected in red, and dead cells in green. In an automated process multiple images of each sample were taken and analyzed with a plugin for the image analysis software ImageJ to identify dead (green), non-producing (red) and producing (orange) cells. The results of the presented validation experiments revealed a good identification with 98 % red cells in the wt sample and 90 % orange cells in the producer sample. The detected wt pigmentation phenotype (red cells) in the producer sample were either not fully induced yet (in 48 h induced

  17. Photophysiological and photosynthetic complex changes during iron starvation in Synechocystis sp. PCC 6803 and Synechococcus elongatus PCC 7942.

    Directory of Open Access Journals (Sweden)

    Jared M Fraser

    Full Text Available Iron is an essential component in many protein complexes involved in photosynthesis, but environmental iron availability is often low as oxidized forms of iron are insoluble in water. To adjust to low environmental iron levels, cyanobacteria undergo numerous changes to balance their iron budget and mitigate the physiological effects of iron depletion. We investigated changes in key protein abundances and photophysiological parameters in the model cyanobacteria Synechococcus PCC 7942 and Synechocystis PCC 6803 over a 120 hour time course of iron deprivation. The iron stress induced protein (IsiA accumulated to high levels within 48 h of the onset of iron deprivation, reaching a molar ratio of ~42 IsiA : Photosystem I in Synechococcus PCC 7942 and ~12 IsiA : Photosystem I in Synechocystis PCC 6803. Concomitantly the iron-rich complexes Cytochrome b6f and Photosystem I declined in abundance, leading to a decrease in the Photosystem I : Photosystem II ratio. Chlorophyll fluorescence analyses showed a drop in electron transport per Photosystem II in Synechococcus, but not in Synechocystis after iron depletion. We found no evidence that the accumulated IsiA contributes to light capture by Photosystem II complexes.

  18. Composición bioquímica y toxicidad de colonias comestibles de la cianobacteria andina Nostoc sp. Llayta

    OpenAIRE

    Galetovic, Alexandra; E. Araya, Jorge; Gómez-Silva, Benito

    2017-01-01

    RESUMEN Llayta es el nombre asignado a la biomasa seca de macrocolonias de una cianobacteria filamentosa, clasificada como Nostoc sp. Llayta crece en humedales andinos de Sudamérica y ha sido considerada como un ingrediente alimenticio desde tiempos precolombinos. Este estudio pretende contribuir a la definición de la calidad nutritiva de la Llayta, proporcionando información sobre su composición bioquímica e identificar aspectos críticos de interés toxicológico. Los resultados mostraron que ...

  19. Effects of light intensity on components and topographical structures of extracellular polysaccharides from the cyanobacteria Nostoc sp.

    Science.gov (United States)

    Ge, Hongmei; Xia, Ling; Zhou, Xuping; Zhang, Delu; Hu, Chunxiang

    2014-02-01

    A study on the effects of light intensity (40 and 80 μE/m(2)/sec) on the components and topographical structures of extracellular polysaccharides (EPS) was carried out in cyanobacteria Nostoc sp.. EPS yield increased with light intensity. However, light intensity did not significantly affect the EPS fractions and monosaccharide composition. Higher light intensity generally resulted in higher protein content of EPS in similar fractions. The topographical structure of EPS, investigated by atomic force microscopy, appeared as spherical lumps, chains and networks. The long chains were observed at higher light intensity. Thus, light intensity affected the yield and nature of EPS.

  20. Synechocystis sp. PCC 6803 CruA (sll0147) encodes lycopene cyclase and requires bound chlorophyll a for activity.

    Science.gov (United States)

    Xiong, Wei; Shen, Gaozhong; Bryant, Donald A

    2017-03-01

    The genome of the model cyanobacterium, Synechococcus sp. PCC 7002, encodes two paralogs of CruA-type lycopene cyclases, SynPCC7002_A2153 and SynPCC7002_A0043, which are denoted cruA and cruP, respectively. Unlike the wild-type strain, a cruA deletion mutant is light-sensitive, grows slowly, and accumulates lycopene, γ-carotene, and 1-OH-lycopene; however, this strain still produces β-carotene and other carotenoids derived from it. Expression of cruA from Synechocystis sp. PCC 6803 (cruA 6803 ) in Escherichia coli strains that synthesize either lycopene or γ-carotene did not lead to the synthesis of either γ-carotene or β-carotene, respectively. However, expression of this orthologous cruA 6803 gene (sll0147) in the Synechococcus sp. PCC 7002 cruA deletion mutant produced strains with phenotypic properties identical to the wild type. CruA 6803 was purified from Synechococcus sp. PCC 7002 by affinity chromatography, and the purified protein was pale yellow-green due to the presence of bound chlorophyll (Chl) a and β-carotene. Native polyacrylamide gel electrophoresis of the partly purified protein in the presence of lithium dodecylsulfate at 4 °C confirmed that the protein was yellow-green in color. When purified CruA 6803 was assayed in vitro with either lycopene or γ-carotene as substrate, β-carotene was synthesized. These data establish that CruA 6803 is a lycopene cyclase and that it requires a bound Chl a molecule for activity. Possible binding sites for Chl a and the potential regulatory role of the Chl a in coordination of Chl and carotenoid biosynthesis are discussed.

  1. In vivo assessment of the hepatotoxicity of a new Nostoc isolate from the Nile River: Nostoc sp. strain NRI.

    Science.gov (United States)

    Abu-Serie, Marwa M; Nasser, Nermine; Abd El-Wahab, Abeer; Shehawy, Rehab; Pienaar, Harrison; Baddour, Nahed; Amer, Ranya

    2018-03-01

    Nostoc sp. is one of the most widely distributed cyanobacterial genera that produce potentially protein phosphatase (PP) inhibitor; microcystins (MCs). MCs have posed a worldwide concern due to predominant hepatotoxicity to human health. We have previously isolated a Nostoc strain (NR1) from the Nile River (the main water supply in Egypt) and this strain exerted production of rare and highly toxic MC; demethylated microcystin-LR. There is no data concerning risk factors of liver diseases for human and animal exposure to NR1-contaminated drinking water yet. It is thus important to evaluate acute (LD 50 dose), subacute (0.01% and 10% of LD 50 dose) and subchronic (0.01% and 10% of LD 50 dose) hepatotoxicity's NR1 extract using experimental mice. Mice groups, who orally received 0.01% LD 50 , represented a permissible concentration of the World Health Organization (WHO) for MC in drinking water. Several parameters were detected, including hepatotoxicity (i.e. PP activity, liver function, oxidative stress markers and DNA fragmentation), pro-inflammatory cytokine (TNF-α) and liver histopathology. Our results demonstrated LD 50 of NR1 extract was at 15,350 mg/kg body weight and caused hepatotoxicity that attributed to PP inhibition and a significant increase of hepatic damage biomarkers with lipid accumulation. Moreover, NR1 extract induced hepatic oxidative damage that may have led to DNA fragmentation and production of TNF-α. As demonstrated from the histopathological study, NR1 extract caused a severe collapse of cytoskeleton with subsequent focal degeneration of hepatocytes, necroinflammation and steatosis. The grade of hepatotoxicity in subacute (10% of LD 50 ) group was higher than that in the subchronic (10% of LD 50 and 0.01% of LD 50 , WHOch, respectively) groups. No significant hepatotoxicity was detectable for subacute (0.01% of LD 50 , WHOac) group. NR1 is therefore considered as one of the harmful and life-threatening cyanobacteria for Egyptian people

  2. Dry heat tolerance of the dry colony in Nostoc sp. HK-01 for useful usage in space agriculture

    Science.gov (United States)

    Kimura, Shunta; Tomita-Yokotani, Kaori; Yamashita, Masamichi; Sato, Seigo; Katoh, Hiroshi

    Space agriculture producing foods is important as one of approach for space habitation. Nostoc sp. HK-01 is one of terrestrial cyanobacterium having a high dry tolerance and it has several ability, photosynthesis, nitrogen fixation and usefulness as a food, it is thought that it can be used for space agriculture. Besides, a study on each tolerance predicted at the time of introduction to space agriculture is necessary. Therefore, as one of the tolerance that are intended to space environment, dry heat ( 100(°) C, 10 h ) tolerance of dry colony in Nostoc sp. HK-01 has been investigated, but the detail function of them has not yet been elucidated. We focused on the extracellular polysaccharides ( EPS ) having the various tolerance, desiccation, low temperature, NaCl, and heavy particle beam. We will consider the function and useful usage of this cyanobacterum in space agriculture after the consideration of the results of contribution of the possibility that EPS improves dry heat tolerance under a dry condition.

  3. Improved Eco-Friendly Recombinant Anabaena sp. Strain PCC7120 with Enhanced Nitrogen Biofertilizer Potential▿

    Science.gov (United States)

    Chaurasia, Akhilesh Kumar; Apte, Shree Kumar

    2011-01-01

    Photosynthetic, nitrogen-fixing Anabaena strains are native to tropical paddy fields and contribute to the carbon and nitrogen economy of such soils. Genetic engineering was employed to improve the nitrogen biofertilizer potential of Anabaena sp. strain PCC7120. Constitutive enhanced expression of an additional integrated copy of the hetR gene from a light-inducible promoter elevated HetR protein expression and enhanced functional heterocyst frequency in the recombinant strain. The recombinant strain displayed consistently higher nitrogenase activity than the wild-type strain and appeared to be in homeostasis with compatible modulation of photosynthesis and respiration. The enhanced combined nitrogen availability from the recombinant strain positively catered to the nitrogen demand of rice seedlings in short-term hydroponic experiments and supported better growth. The engineered strain is stable, eco-friendly, and useful for environmental application as nitrogen biofertilizer in paddy fields. PMID:21057013

  4. Improved eco-friendly recombinant Anabaena sp. strain PCC7120 with enhanced nitrogen biofertilizer potential.

    Science.gov (United States)

    Chaurasia, Akhilesh Kumar; Apte, Shree Kumar

    2011-01-01

    Photosynthetic, nitrogen-fixing Anabaena strains are native to tropical paddy fields and contribute to the carbon and nitrogen economy of such soils. Genetic engineering was employed to improve the nitrogen biofertilizer potential of Anabaena sp. strain PCC7120. Constitutive enhanced expression of an additional integrated copy of the hetR gene from a light-inducible promoter elevated HetR protein expression and enhanced functional heterocyst frequency in the recombinant strain. The recombinant strain displayed consistently higher nitrogenase activity than the wild-type strain and appeared to be in homeostasis with compatible modulation of photosynthesis and respiration. The enhanced combined nitrogen availability from the recombinant strain positively catered to the nitrogen demand of rice seedlings in short-term hydroponic experiments and supported better growth. The engineered strain is stable, eco-friendly, and useful for environmental application as nitrogen biofertilizer in paddy fields.

  5. A bioelectrochemical approach to characterize extracellular electron transfer by Synechocystis sp. PCC6803.

    Directory of Open Access Journals (Sweden)

    Angelo Cereda

    Full Text Available Biophotovoltaic devices employ photosynthetic organisms at the anode of a microbial fuel cell to generate electrical power. Although a range of cyanobacteria and algae have been shown to generate photocurrent in devices of a multitude of architectures, mechanistic understanding of extracellular electron transfer by phototrophs remains minimal. Here we describe a mediatorless bioelectrochemical device to measure the electrogenic output of a planktonically grown cyanobacterium, Synechocystis sp. PCC6803. Light dependent production of current is measured, and its magnitude is shown to scale with microbial cell concentration and light intensity. Bioelectrochemical characterization of a Synechocystis mutant lacking Photosystem II demonstrates conclusively that production of the majority of photocurrent requires a functional water splitting aparatus and electrons are likely ultimately derived from water. This shows the potential of the device to rapidly and quantitatively characterize photocurrent production by genetically modified strains, an approach that can be used in future studies to delineate the mechanisms of cyanobacterial extracellular electron transport.

  6. An integrative approach to energy, carbon, and redox metabolism in the cyanobacterium Synechocystis sp. PCC 6803

    Energy Technology Data Exchange (ETDEWEB)

    Overbeek, Ross; Fonstein, Veronika; Osterman, Andrei; Gerdes, Svetlana; Vassieva, Olga; Zagnitko, Olga; Rodionov, Dmitry

    2005-02-15

    The team of the Fellowship for Interpretation of Genomes (FIG) under the leadership of Ross Overbeek, began working on this Project in November 2003. During the previous year, the Project was performed at Integrated Genomics Inc. A transition from the industrial environment to the public domain prompted us to adjust some aspects of the Project. Notwithstanding the challenges, we believe that these adjustments had a strong positive impact on our deliverables. Most importantly, the work of the research team led by R. Overbeek resulted in the deployment of a new open source genomic platform, the SEED (Specific Aim 1). This platform provided a foundation for the development of CyanoSEED a specialized portal to comparative analysis and metabolic reconstruction of all available cyanobacterial genomes (Specific Aim 3). The SEED represents a new generation of software for genome analysis. Briefly, it is a portable and extendable system, containing one of the largest and permanently growing collections of complete and partial genomes. The complete system with annotations and tools is freely available via browsing or via installation on a user's Mac or Linux computer. One of the important unique features of the SEED is the support of metabolic reconstruction and comparative genome analysis via encoding and projection of functional subsystems. During the project period, the FIG research team has validated the new software by developing a significant number of core subsystems, covering many aspects of central metabolism (Specific Aim 2), as well as metabolic areas specific for cyanobacteria and other photoautotrophic organisms (Specific Aim 3). In addition to providing a proof of technology and a starting point for further community-based efforts, these subsystems represent a valuable asset. An extensive coverage of central metabolism provides the bulk of information required for metabolic modeling in Synechocystis sp.PCC 6803. Detailed analysis of several subsystems

  7. Unravelling the cross-talk between iron starvation and oxidative stress responses highlights the key role of PerR (alr0957) in peroxide signalling in the cyanobacterium Nostoc PCC 7120.

    Science.gov (United States)

    Yingping, Fan; Lemeille, Sylvain; Talla, Emmanuel; Janicki, Annick; Denis, Yann; Zhang, Cheng-Cai; Latifi, Amel

    2014-10-01

    The cyanobacterial phylum includes oxygenic photosynthetic prokaryotes of a wide variety of morphologies, metabolisms and ecologies. Their adaptation to their various ecological niches is mainly achieved by sophisticated regulatory mechanisms and depends on a fine cross-talk between them. We assessed the global transcriptomic response of the filamentous cyanobacterium Nostoc PCC 7120 to iron starvation and oxidative stress. More than 20% of the differentially expressed genes in response to iron stress were also responsive to oxidative stress. These transcripts include antioxidant proteins-encoding genes that confirms that iron depletion leads to reactive oxygen accumulation. The activity of the Fe-superoxide dismutase was not significantly decreased under iron starvation, indicating that the oxidative stress generated under iron deficiency is not a consequence of (SOD) deficiency. The transcriptional data indicate that the adaptation of Nostoc to iron-depleted conditions displays important differences with what has been shown in unicellular cyanobacteria. While the FurA protein that regulates the response to iron deprivation has been well characterized in Nostoc, the regulators in charge of the oxidative stress response are unknown. Our study indicates that the alr0957 (perR) gene encodes the master regulator of the peroxide stress. PerR is a peroxide-sensor repressor that senses peroxide by metal-catalysed oxidation.

  8. Network analysis of transcriptomics expands regulatory landscapes in Synechococcus sp. PCC 7002

    Energy Technology Data Exchange (ETDEWEB)

    McClure, Ryan S.; Overall, Christopher C.; McDermott, Jason E.; Hill, Eric A.; Markillie, Lye Meng; McCue, Lee Ann; Taylor, Ronald C.; Ludwig, Marcus; Bryant, Donald A.; Beliaev, Alexander S.

    2016-08-27

    Cyanobacterial regulation of gene expression must contend with a genome organization that lacks apparent functional context, as the majority of cellular processes and metabolic pathways are encoded by genes found at disparate locations across the genome. In addition, the fact that coordinated regulation of cyanobacterial cellular machinery takes place with significantly fewer transcription factors, compared to other Eubacteria, suggests the involvement of post-transcriptional mechanisms and regulatory adaptations which are not fully understood. Global transcript abundance from model cyanobacterium Synechococcus sp. PCC 7002 grown under 42 different conditions was analyzed using context-likelihood of relatedness. The resulting 903-gene network, which was organized into 11 modules, not only allowed classification of cyanobacterial responses to specific environmental variables but provided insight into the transcriptional network topology and led to the expansion of predicted regulons. When used in conjunction with genome sequence, the global transcript abundance allowed identification of putative post-transcriptional changes in expression as well as novel potential targets of both DNA binding proteins and asRNA regulators. The results offer a new perspective into the multi-level regulation that governs cellular adaptations of fast-growing physiologically robust cyanobacterium Synechococcus sp. PCC 7002 to changing environmental variables. It also extends a methodological knowledge-based framework for studying multi-scale regulatory mechanisms that operate in cyanobacteria. Finally, it provides valuable context for integrating systems-level data to enhance evidence-driven genomic annotation, especially in organisms where traditional context analyses cannot be implemented due to lack of operon-based functional organization.

  9. Analysis of carbohydrate storage granules in the diazotrophic cyanobacterium Cyanothece sp. PCC 7822

    Energy Technology Data Exchange (ETDEWEB)

    Welkie, David G. [Purdue Univ., West Lafayette, IN (United States); Sherman, Debra M. [Purdue Univ., West Lafayette, IN (United States); Chrisler, William B. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Orr, Galya [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Sherman, Louis A. [Purdue Univ., West Lafayette, IN (United States)

    2013-10-19

    The unicellular diazotrophic cyanobacteria of the genus Cyanothece demonstrate oscillations in nitrogenase activity and H2 production when grown under 12h light-12h dark cycles. We established that Cyanothece sp. PCC 7822 allows for the construction of knock-out mutants and our objective was to improve the growth characteristics of this strain and to identify the nature of the intracellular storage granules. We report the physiological and morphological effects of reduction in nitrate and phosphate concentrations in BG-11 media on this strain. We developed a series of BG-11-derived growth media and monitored batch culture growth, nitrogenase activity and nitrogenase-mediated hydrogen production, culture synchronicity, and intracellular storage content. Reduction in NaNO3 and K2HPO4 concentrations from 17.6 and 0.23 mM to 4.41 and 0.06 mM, respectively, improved growth characteristics such as cell size and uniformity, and enhanced the rate of cell division. Cells grown in this low NP BG-11 were less complex, a parameter that related to the composition of the intracellular storage granules. Cells grown in low NP BG-11 had less polyphosphate, fewer polyhydroxybutyrate granules and many smaller granules became evident. Biochemical analysis and transmission electron microscopy using the histocytochemical PATO technique demonstrated that these small granules contained glycogen. The glycogen levels and the number of granules per cell correlated nicely with a 2.3 to 3.3-fold change from the minimum at L0 to the maximum at D0. The differences in granule morphology and enzymes between Cyanothece ATCC 51142 and Cyanothece PCC 7822 provide insights into the formation of large starch-like granules in some cyanobacteria.

  10. Reconstruction and comparison of the metabolic potential of cyanobacteria Cyanothece sp. ATCC 51142 and Synechocystis sp. PCC 6803.

    Directory of Open Access Journals (Sweden)

    Rajib Saha

    Full Text Available Cyanobacteria are an important group of photoautotrophic organisms that can synthesize valuable bio-products by harnessing solar energy. They are endowed with high photosynthetic efficiencies and diverse metabolic capabilities that confer the ability to convert solar energy into a variety of biofuels and their precursors. However, less well studied are the similarities and differences in metabolism of different species of cyanobacteria as they pertain to their suitability as microbial production chassis. Here we assemble, update and compare genome-scale models (iCyt773 and iSyn731 for two phylogenetically related cyanobacterial species, namely Cyanothece sp. ATCC 51142 and Synechocystis sp. PCC 6803. All reactions are elementally and charge balanced and localized into four different intracellular compartments (i.e., periplasm, cytosol, carboxysome and thylakoid lumen and biomass descriptions are derived based on experimental measurements. Newly added reactions absent in earlier models (266 and 322, respectively span most metabolic pathways with an emphasis on lipid biosynthesis. All thermodynamically infeasible loops are identified and eliminated from both models. Comparisons of model predictions against gene essentiality data reveal a specificity of 0.94 (94/100 and a sensitivity of 1 (19/19 for the Synechocystis iSyn731 model. The diurnal rhythm of Cyanothece 51142 metabolism is modeled by constructing separate (light/dark biomass equations and introducing regulatory restrictions over light and dark phases. Specific metabolic pathway differences between the two cyanobacteria alluding to different bio-production potentials are reflected in both models.

  11. Utility of Synechocystis sp. PCC glutaredoxin A as a platform to study high-resolution mutagenesis of proteins.

    Directory of Open Access Journals (Sweden)

    David B Knaff

    2013-11-01

    Full Text Available Glutaredoxin from the cyanobacterium Synechocystis sp. PCC 6803 is a small protein, containing only 88 amino acids, that participates in a large number of redox reactions, serving both as an electron donor for enzyme-catalyzed reductions and as a regulator of diverse metabolic pathways. The crystal structures of glutaredoxins from several species have been solved, including the glutaredoxin A isoform from the cyanobacterium Synechocystis sp. PCC 6803. We have utilized the small size of Synechocystis glutaredoxin A and its propensity to form protein crystals that diffract to high resolution to explore a long-standing question in biochemistry; i.e., what are the effects of mutations on protein structure and function? Taking advantage of these properties, we have initiated a long-term educational project that would examine the structural and biochemical changes in glutaredoxin as a function of single-point mutational replacements. Here, we report some of the mutational effects that we have observed to date.

  12. Lack of Phosphatidylglycerol Inhibits Chlorophyll Biosynthesis at Multiple Sites and Limits Chlorophyllide Reutilization in Synechocystis sp Strain PCC 6803

    Czech Academy of Sciences Publication Activity Database

    Kopečná, Jana; Pilný, Jan; Krynická, Vendula; Tomčala, Aleš; Kis, M.; Gombos, Z.; Komenda, Josef; Sobotka, Roman

    2015-01-01

    Roč. 169, č. 2 (2015), s. 1307-1317 ISSN 0032-0889 R&D Projects: GA MŠk LO1416; GA MŠk EE2.3.30.0059; GA ČR GBP501/12/G055 Institutional support: RVO:61388971 ; RVO:60077344 Keywords : II REACTION-CENTER * PHOTOSYSTEM-II * SP PCC-6803 Subject RIV: CE - Biochemistry Impact factor: 6.280, year: 2015

  13. Resistance to the photosystem II herbicide diuron is dominant to sensitivity in the cyanobacterium Synechococcus sp. PCC7942

    OpenAIRE

    Brusslan, Judy; Haselkorn, Robert

    1989-01-01

    The transformable cyanobacterium, Synechococcus sp. PCC7942, was used to study the genetics of resistance to the herbicide diuron. In wild-type cells, diuron binds to one of the core proteins, called D1, of photosystem II reaction centres. This binding prevents the transfer of electrons from QA, the primary quinone acceptor, to QB, which is necessary to create the charge separation that drives ATP synthesis. A single amino acid substitution in the D1 protein reduces diuron binding and confers...

  14. Construction of new synthetic biology tools for the control of gene expression in the cyanobacterium Synechococcus sp. strain PCC 7002.

    Science.gov (United States)

    Zess, Erin K; Begemann, Matthew B; Pfleger, Brian F

    2016-02-01

    Predictive control of gene expression is an essential tool for developing synthetic biological systems. The current toolbox for controlling gene expression in cyanobacteria is a barrier to more in-depth genetic analysis and manipulation. Towards relieving this bottleneck, this work describes the use of synthetic biology to construct an anhydrotetracycline-based induction system and adapt a trans-acting small RNA (sRNA) system for use in the cyanobacterium Synechococcus sp. strain PCC 7002. An anhydrotetracycline-inducible promoter was developed to maximize intrinsic strength and dynamic range. The resulting construct, PEZtet , exhibited tight repression and a maximum 32-fold induction upon addition of anhydrotetracycline. Additionally, a sRNA system based on the Escherichia coli IS10 RNA-IN/OUT regulator was adapted for use in Synechococcus sp. strain PCC 7002. This system exhibited 70% attenuation of target gene expression, providing a demonstration of the use of sRNAs for differential gene expression in cyanobacteria. These systems were combined to produce an inducible sRNA system, which demonstrated 59% attenuation of target gene expression. Lastly, the role of Hfq, a critical component of sRNA systems in E. coli, was investigated. Genetic studies showed that the Hfq homolog in Synechococcus sp. strain PCC 7002 did not impact repression by the engineered sRNA system. In summary, this work describes new synthetic biology tools that can be applied to physiological studies, metabolic engineering, or sRNA platforms in Synechococcus sp. strain PCC 7002. © 2015 Wiley Periodicals, Inc.

  15. Mutation of the murC and murB Genes Impairs Heterocyst Differentiation in Anabaena sp. Strain PCC 7120

    OpenAIRE

    Videau, Patrick; Rivers, Orion S.; Ushijima, Blake; Oshiro, Reid T.; Kim, Min Joo; Philmus, Benjamin; Cozy, Loralyn M.

    2016-01-01

    To stabilize cellular integrity in the face of environmental perturbations, most bacteria, including cyanobacteria, synthesize and maintain a strong, flexible, three-dimensional peptidoglycan lattice. Anabaena sp. strain PCC 7120 is a filamentous cyanobacterium capable of differentiating morphologically distinct nitrogen-fixing heterocyst cells in a periodic pattern. While heterocyst development has been shown to require proper peptidoglycan remodeling, the role of peptidoglycan synthesis has...

  16. An Integrative Approach to Energy, Carbon, and Redox Metabolism in the Cyanobacterium Synechocystis sp. PCC 6803. Special Report

    Energy Technology Data Exchange (ETDEWEB)

    Overbeek, R.

    2003-06-30

    The main objectives for the first year were to produce a detailed metabolic reconstruction of synechocystis sp. PCC 6803 especially in interrelated areas of photosynthesis, respiration, and central carbon metabolism to support a more complete understanding and modeling of this organism. Additionally, Integrated Genomics, Inc., provided detailed bioinformatic analysis of selected functional systems related to carbon and energy generation and utilization, and of the corresponding pathways, functional roles and individual genes to support wet lab experiments by collaborators.

  17. A Cluster of Five Genes Essential for the Utilization of Dihydroxamate Xenosiderophores in Synechocystis sp. PCC 6803.

    Science.gov (United States)

    Obando S, Tobias A; Babykin, Michael M; Zinchenko, Vladislav V

    2018-05-21

    The unicellular freshwater cyanobacterium Synechocystis sp. PCC 6803 is capable of using dihydroxamate xenosiderophores, either ferric schizokinen (FeSK) or a siderophore of the filamentous cyanobacterium Anabaena variabilis ATCC 29413 (SAV), as the sole source of iron in the TonB-dependent manner. The fecCDEB1-schT gene cluster encoding a siderophore transport system that is involved in the utilization of FeSK and SAV in Synechocystis sp. PCC 6803 was identified. The gene schT encodes TonB-dependent outer membrane transporter, whereas the remaining four genes encode the ABC-type transporter FecB1CDE formed by the periplasmic binding protein FecB1, the transmembrane permease proteins FecC and FecD, and the ATPase FecE. Inactivation of any of these genes resulted in the inability of cells to utilize FeSK and SAV. Our data strongly suggest that Synechocystis sp. PCC 6803 can readily internalize Fe-siderophores via the classic TonB-dependent transport system.

  18. Improved sugar-free succinate production by Synechocystis sp. PCC 6803 following identification of the limiting steps in glycogen catabolism

    Directory of Open Access Journals (Sweden)

    Tomohisa Hasunuma

    2016-12-01

    Full Text Available Succinate produced by microorganisms can replace currently used petroleum-based succinate but typically requires mono- or poly-saccharides as a feedstock. The cyanobacterium Synechocystis sp. PCC6803 can produce organic acids such as succinate from CO2 not supplemented with sugars under dark anoxic conditions using an unknown metabolic pathway. The TCA cycle in cyanobacteria branches into oxidative and reductive routes. Time-course analyses of the metabolome, transcriptome and metabolic turnover described here revealed dynamic changes in the metabolism of Synechocystis sp. PCC6803 cultivated under dark anoxic conditions, allowing identification of the carbon flow and rate-limiting steps in glycogen catabolism. Glycogen biosynthesized from CO2 assimilated during periods of light exposure is catabolized to succinate via glycolysis, the anaplerotic pathway, and the reductive TCA cycle under dark anoxic conditions. Expression of the phosphoenolpyruvate (PEP carboxylase gene (ppc was identified as a rate-limiting step in succinate biosynthesis and this rate limitation was alleviated by ppc overexpression, resulting in improved succinate excretion. The sugar-free succinate production was further enhanced by the addition of bicarbonate. In vivo labeling with NaH13CO3 clearly showed carbon incorporation into succinate via the anaplerotic pathway. Bicarbonate is in equilibrium with CO2. Succinate production by Synechocystis sp. PCC6803 therefore holds significant promise for CO2 capture and utilization. Keywords: Autofermentation, Cyanobacteria, Dynamic metabolic profiling, Metabolomics, Succinate, Synechocystis

  19. Biocomputional construction of a gene network under acid stress in Synechocystis sp. PCC 6803.

    Science.gov (United States)

    Li, Yi; Rao, Nini; Yang, Feng; Zhang, Ying; Yang, Yang; Liu, Han-ming; Guo, Fengbiao; Huang, Jian

    2014-01-01

    Acid stress is one of the most serious threats that cyanobacteria have to face, and it has an impact at all levels from genome to phenotype. However, very little is known about the detailed response mechanism to acid stress in this species. We present here a general analysis of the gene regulatory network of Synechocystis sp. PCC 6803 in response to acid stress using comparative genome analysis and biocomputational prediction. In this study, we collected 85 genes and used them as an initial template to predict new genes through co-regulation, protein-protein interactions and the phylogenetic profile, and 179 new genes were obtained to form a complete template. In addition, we found that 11 enriched pathways such as glycolysis are closely related to the acid stress response. Finally, we constructed a regulatory network for the intricate relationship of these genes and summarize the key steps in response to acid stress. This is the first time a bioinformatic approach has been taken systematically to gene interactions in cyanobacteria and the elaboration of their cell metabolism and regulatory pathways under acid stress, which is more efficient than a traditional experimental study. The results also provide theoretical support for similar research into environmental stresses in cyanobacteria and possible industrial applications. Copyright © 2014 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  20. Characterization of three bioenergetically active respiratory terminal oxidases in the cyanobacterium Synechocystis sp. strain PCC 6803.

    Science.gov (United States)

    Pils, D; Schmetterer, G

    2001-09-25

    Synechocystis sp. PCC 6803 contains three respiratory terminal oxidases (RTOs): cytochrome c oxidase (Cox), quinol oxidase (Cyd), and alternate RTO (ARTO). Mutants lacking combinations of the RTOs were used to characterize these key enzymes of respiration. Pentachlorophenol and 2-heptyl-4-hydroxy-quinoline-N-oxide inhibited Cyd completely, but had little effect on electron transport to the other RTOs. KCN inhibited all three RTOs but the in vivo K(I) for Cox and Cyd was quite different (7 vs. 27 microM), as was their affinity for oxygen (K(M) 1.0 vs. 0.35 microM). ARTO has a very low respiratory activity. However, when uptake of 3-O-methylglucose, an active H+ co-transport, was used to monitor energization of the cytoplasmic membrane, ARTO was similarly effective as the other RTOs. As removal of the gene for cytochrome c(553) had the same effects as removal of ARTO genes, we propose that the ARTO might be a second Cox. The possible functions, localization and regulation of the RTOs are discussed.

  1. Monitoring Photosynthesis in Individual Cells of Synechocystis sp. PCC 6803 on a Picosecond Timescale

    Science.gov (United States)

    Krumova, S.B.; Laptenok, S.P.; Borst, J.W.; Ughy, B.; Gombos, Z.; Ajlani, G.; van Amerongen, H.

    2010-01-01

    Picosecond fluorescence kinetics of wild-type (WT) and mutant cells of Synechocystis sp. PCC 6803, were studied at the ensemble level with a streak-camera and at the cell level using fluorescence-lifetime-imaging microscopy (FLIM). The FLIM measurements are in good agreement with the ensemble measurements, but they (can) unveil variations between and within cells. The BE mutant cells, devoid of photosystem II (PSII) and of the light-harvesting phycobilisomes, allowed the study of photosystem I (PSI) in vivo for the first time, and the observed 6-ps equilibration process and 25-ps trapping process are the same as found previously for isolated PSI. No major differences are detected between different cells. The PAL mutant cells, devoid of phycobilisomes, show four lifetimes: ∼20 ps (PSI and PSII), ∼80 ps, ∼440 ps, and 2.8 ns (all due to PSII), but not all cells are identical and variations in the kinetics are traced back to differences in the PSI/PSII ratio. Finally, FLIM measurements on WT cells reveal that in some cells or parts of cells, phycobilisomes are disconnected from PSI/PSII. It is argued that the FLIM setup used can become instrumental in unraveling photosynthetic regulation mechanisms in the future. PMID:20858447

  2. Physical, chemical, and metabolic state sensors expand the synthetic biology toolbox for Synechocystis sp. PCC 6803.

    Science.gov (United States)

    Immethun, Cheryl M; DeLorenzo, Drew M; Focht, Caroline M; Gupta, Dinesh; Johnson, Charles B; Moon, Tae Seok

    2017-07-01

    Many under-developed organisms possess important traits that can boost the effectiveness and sustainability of microbial biotechnology. Photoautotrophic cyanobacteria can utilize the energy captured from light to fix carbon dioxide for their metabolic needs while living in environments not suited for growing crops. Various value-added compounds have been produced by cyanobacteria in the laboratory; yet, the products' titers and yields are often not industrially relevant and lag behind what have been accomplished in heterotrophic microbes. Genetic tools for biological process control are needed to take advantage of cyanobacteria's beneficial qualities, as tool development also lags behind what has been created in common heterotrophic hosts. To address this problem, we developed a suite of sensors that regulate transcription in the model cyanobacterium Synechocystis sp. PCC 6803 in response to metabolically relevant signals, including light and the cell's nitrogen status, and a family of sensors that respond to the inexpensive chemical, l-arabinose. Increasing the number of available tools enables more complex and precise control of gene expression. Expanding the synthetic biology toolbox for this cyanobacterium also improves our ability to utilize this important under-developed organism in biotechnology. Biotechnol. Bioeng. 2017;114: 1561-1569. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  3. Effect of culture density on biomass production and light utilization efficiency of Synechocystis sp. PCC 6803.

    Science.gov (United States)

    Straka, Levi; Rittmann, Bruce E

    2018-02-01

    The viability of large-scale microalgae cultivation depends on providing optimal growth conditions, for which a key operational parameter is culture density. Using Synechocystis sp. PCC 6803, we conducted a series of fixed-density, steady-state experiments and one batch-growth experiment to investigate the role of culture density on biomass production and light utilization efficiency. In all cases, the fixed-density, steady-state experiments and batch-growth experiment showed good agreement. The highest biomass production rates (260 mg L -1  d -1 ) and efficiency for converting light energy to biomass (0.80 μg (μmol photons) -1 ) occurred together at a culture density near 760 mg L -1 , which approximately corresponded to the lowest culture density where almost all incident light was absorbed. The ratio of OD 680 /OD 735 increased with culture density up to the point of maximum productivity, where it plateaued (at a value of 2.4) for higher culture densities. This change in OD 680 /OD 735 indicates a photoacclimation effect that depended on culture density. Very high culture densities led to a sharp decline in efficiency of biomass production per photons absorbed, likely due to a combination of increased decay relative to growth, metabolic changes due to cell-cell interactions, and photodamage due to mixing between regions with high light intensity and zero light intensity. © 2017 Wiley Periodicals, Inc.

  4. Genomic responses to arsenic in the cyanobacterium Synechocystis sp. PCC 6803.

    Directory of Open Access Journals (Sweden)

    Ana María Sánchez-Riego

    Full Text Available Arsenic is a ubiquitous contaminant and a toxic metalloid which presents two main redox states in nature: arsenite [As(III] and arsenate [As(V]. Arsenic resistance in Synechocystis sp. strain PCC 6803 is mediated by the arsBHC operon and two additional arsenate reductases encoded by the arsI1 and arsI2 genes. Here we describe the genome-wide responses to the presence of arsenate and arsenite in wild type and mutants in the arsenic resistance system. Both forms of arsenic produced similar responses in the wild type strain, including induction of several stress related genes and repression of energy generation processes. These responses were transient in the wild type strain but maintained in time in an arsB mutant strain, which lacks the arsenite transporter. In contrast, the responses observed in a strain lacking all arsenate reductases were somewhat different and included lower induction of genes involved in metal homeostasis and Fe-S cluster biogenesis, suggesting that these two processes are targeted by arsenite in the wild type strain. Finally, analysis of the arsR mutant strain revealed that ArsR seems to only control 5 genes in the genome. Furthermore, the arsR mutant strain exhibited hypersentivity to nickel, copper and cadmium and this phenotype was suppressed by mutation in arsB but not in arsC gene suggesting that overexpression of arsB is detrimental in the presence of these metals in the media.

  5. Metabolic flux analysis of the hydrogen production potential in Synechocystis sp. PCC6803

    Energy Technology Data Exchange (ETDEWEB)

    Navarro, E. [Departamento de Lenguajes y Ciencias de la Computacion, Campus de Teatrinos, Universidad de Malaga, 29071 Malaga (Spain); Montagud, A.; Fernandez de Cordoba, P.; Urchueguia, J.F. [Instituto Universitario de Matematica Pura y Aplicada, Universidad Politecnica de Valencia, Camino de Vera 14, 46022 Valencia (Spain)

    2009-11-15

    Hydrogen is a promising energy vector; however, finding methods to produce it from renewable sources is essential to allow its wide-scale use. In that line, biological hydrogen production, although it is considered as a possible alternative, requires substantial improvements to overcome its present low yields. In that direction, genetic manipulation probably will play a central role and from that point of view metabolic flux analysis (MFA) constitutes an important tool to guide a priori most suitable genetic modifications oriented to a hydrogen yield increase. In this work MFA has been applied to analyze hydrogen photoproduction of Synechocystis sp. PCC6803. Flux analysis was carried out based on literature data and several basic fluxes were estimated in different growing conditions of the system. From this analysis, an upper limit for hydrogen photoproduction has been determined indicating a wide margin for improvement. MFA was also used to find a feasible operating space for hydrogen production, which avoids oxygen inhibition, one of the most important limitations to make hydrogen production cost effective. In addition, a set of biotechnological strategies are proposed that would be consistent with the performed mathematical analysis. (author)

  6. Production of γ-linolenic acid and stearidonic acid by Synechococcus sp. PCC7002 containing cyanobacterial fatty acid desaturase genes

    Science.gov (United States)

    Dong, Xuewei; He, Qingfang; Peng, Zhenying; Yu, Jinhui; Bian, Fei; Li, Youzhi; Bi, Yuping

    2016-07-01

    Genetic modification is useful for improving the nutritional qualities of cyanobacteria. To increase the total unsaturated fatty acid content, along with the ratio of ω-3/ω-6 fatty acids, genetic engineering can be used to modify fatty acid metabolism. Synechococcus sp. PCC7002, a fast-growing cyanobacterium, does not contain a Δ6 desaturase gene and is therefore unable to synthesize γ-linolenic acid (GLA) and stearidonic acid (SDA), which are important in human health. In this work, we constructed recombinant vectors Syd6D, Syd15D and Syd6Dd15D to express the Δ15 desaturase and Δ6 desaturase genes from Synechocystis PCC6803 in Synechococcus sp. PCC7002, with the aim of expressing polyunsaturated fatty acids. Overexpression of the Δ15 desaturase gene in Synechococcus resulted in 5.4 times greater accumulation of α-linolenic acid compared with the wild-type while Δ6 desaturase gene expression produced both GLA and SDA. Co-expression of the two genes resulted in low-level accumulation of GLA but much larger amounts of SDA, accounting for as much to 11.64% of the total fatty acid content.

  7. Development of a high-frequency in vivo transposon mutagenesis system for Synechocystis sp. PCC 6803 and Synechococcus elongatus PCC 7942.

    Science.gov (United States)

    Watabe, Kazuyuki; Mimuro, Mamoru; Tsuchiya, Tohru

    2014-11-01

    Synechocystis sp. PCC 6803 (Synechocystis) is the first sequenced photosynthetic organism and has two advantages: natural transformation and light-activated heterotrophic growth. Such characteristics have mainly promoted reverse genetic analysis in this organism, however, to date approximately 50% of genes are still annotated as 'unknown protein' or 'hypothetical protein'. Therefore, forward genetic analysis is required for the identification of significant genes responsible for photosynthesis and other physiological phenomena among the genes of unknown function. The in vivo transposon mutagenesis system is one of the major methods for random mutagenesis. However, present in vivo transposon mutagenesis systems for cyanobacteria face problems such as relatively low frequency of transposition and repeated transposition in the host cells. In this study, we constructed vectors based on a mini-Tn5-derived vector that was designed to prevent repeated transposition. Our vectors carry a hyperactive transposase and optimized recognition sequence of transposase, which were reported to enhance frequency of transposition. Using the vector, we succeeded in highly frequent transposition (9×10(-3) per recipient cell) in Synechocystis. Transposon insertion sites of 10 randomly selected mutants indicated that the insertion sites spread throughout the genome with low sequence dependency. Furthermore, one of the 10 mutants exhibited the slow-growing phenotype, and the mutant was functionally complemented by using our expression vector. Our system also worked with another model cyanobacterium, Synechococcus elongatus PCC 7942, with high frequency. These results indicate that the developed system can be applied to the forward genetic analysis of a broad range of cyanobacteria. © The Author 2014. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  8. Distinguishing the Roles of Thylakoid Respiratory Terminal Oxidases in the Cyanobacterium Synechocystis sp. PCC 6803.

    Science.gov (United States)

    Ermakova, Maria; Huokko, Tuomas; Richaud, Pierre; Bersanini, Luca; Howe, Christopher J; Lea-Smith, David J; Peltier, Gilles; Allahverdiyeva, Yagut

    2016-06-01

    Various oxygen-utilizing electron sinks, including the soluble flavodiiron proteins (Flv1/3), and the membrane-localized respiratory terminal oxidases (RTOs), cytochrome c oxidase (Cox) and cytochrome bd quinol oxidase (Cyd), are present in the photosynthetic electron transfer chain of Synechocystis sp. PCC 6803. However, the role of individual RTOs and their relative importance compared with other electron sinks are poorly understood, particularly under light. Via membrane inlet mass spectrometry gas exchange, chlorophyll a fluorescence, P700 analysis, and inhibitor treatment of the wild type and various mutants deficient in RTOs, Flv1/3, and photosystem I, we investigated the contribution of these complexes to the alleviation of excess electrons in the photosynthetic chain. To our knowledge, for the first time, we demonstrated the activity of Cyd in oxygen uptake under light, although it was detected only upon inhibition of electron transfer at the cytochrome b6f site and in ∆flv1/3 under fluctuating light conditions, where linear electron transfer was drastically inhibited due to impaired photosystem I activity. Cox is mostly responsible for dark respiration and competes with P700 for electrons under high light. Only the ∆cox/cyd double mutant, but not single mutants, demonstrated a highly reduced plastoquinone pool in darkness and impaired gross oxygen evolution under light, indicating that thylakoid-based RTOs are able to compensate partially for each other. Thus, both electron sinks contribute to the alleviation of excess electrons under illumination: RTOs continue to function under light, operating on slower time ranges and on a limited scale, whereas Flv1/3 responds rapidly as a light-induced component and has greater capacity. © 2016 American Society of Plant Biologists. All Rights Reserved.

  9. Mechanisms of High Temperature Resistance of Synechocystis sp. PCC 6803: An Impact of Histidine Kinase 34

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    Jan Červený

    2015-03-01

    Full Text Available Synechocystis sp. PCC 6803 is a widely used model cyanobacterium for studying responses and acclimation to different abiotic stresses. Changes in transcriptome, proteome, lipidome, and photosynthesis in response to short term heat stress are well studied in this organism, and histidine kinase 34 (Hik34 is shown to play an important role in mediating such response. Corresponding data on long term responses, however, are fragmentary and vary depending on parameters of experiments and methods of data collection, and thus are hard to compare. In order to elucidate how the early stress responses help cells to sustain long-term heat stress, as well as the role of Hik34 in prolonged acclimation, we examined the resistance to long-term heat stress of wild-type and ΔHik34 mutant of Synechocystis. In this work, we were able to precisely control the long term experimental conditions by cultivating Synechocystis in automated photobioreactors, measuring selected physiological parameters within a time range of minutes. In addition, morphological and ultrastructural changes in cells were analyzed and western blotting of individual proteins was used to study the heat stress-affected protein expression. We have shown that the majority of wild type cell population was able to recover after 24 h of cultivation at 44 °C. In contrast, while ΔHik34 mutant cells were resistant to heat stress within its first hours, they could not recover after 24 h long high temperature treatment. We demonstrated that the early induction of HspA expression and maintenance of high amount of other HSPs throughout the heat incubation is critical for successful adaptation to long-term stress. In addition, it appears that histidine kinase Hik34 is an essential component for the long term high temperature resistance.

  10. Mechanisms of High Temperature Resistance of Synechocystis sp. PCC 6803: An Impact of Histidine Kinase 34.

    Science.gov (United States)

    Červený, Jan; Sinetova, Maria A; Zavřel, Tomáš; Los, Dmitry A

    2015-03-02

    Synechocystis sp. PCC 6803 is a widely used model cyanobacterium for studying responses and acclimation to different abiotic stresses. Changes in transcriptome, proteome, lipidome, and photosynthesis in response to short term heat stress are well studied in this organism, and histidine kinase 34 (Hik34) is shown to play an important role in mediating such response. Corresponding data on long term responses, however, are fragmentary and vary depending on parameters of experiments and methods of data collection, and thus are hard to compare. In order to elucidate how the early stress responses help cells to sustain long-term heat stress, as well as the role of Hik34 in prolonged acclimation, we examined the resistance to long-term heat stress of wild-type and ΔHik34 mutant of Synechocystis. In this work, we were able to precisely control the long term experimental conditions by cultivating Synechocystis in automated photobioreactors, measuring selected physiological parameters within a time range of minutes. In addition, morphological and ultrastructural changes in cells were analyzed and western blotting of individual proteins was used to study the heat stress-affected protein expression. We have shown that the majority of wild type cell population was able to recover after 24 h of cultivation at 44 °C. In contrast, while ΔHik34 mutant cells were resistant to heat stress within its first hours, they could not recover after 24 h long high temperature treatment. We demonstrated that the early induction of HspA expression and maintenance of high amount of other HSPs throughout the heat incubation is critical for successful adaptation to long-term stress. In addition, it appears that histidine kinase Hik34 is an essential component for the long term high temperature resistance.

  11. Identification of a transporter Slr0982 involved in ethanol tolerance in cyanobacterium Synechocystis sp. PCC 6803

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    Yanan eZhang

    2015-05-01

    Full Text Available Cyanobacteria have been engineered to produce ethanol through recent synthetic biology efforts. However, one major challenge to the cyanobacterial systems for high-efficiency ethanol production is their low tolerance to the ethanol toxicity. With a major goal to identify novel transporters involved in ethanol tolerance, we constructed gene knockout mutants for 58 transporter-encoding genes of Synechocystis sp. PCC 6803 and screened their tolerance change under ethanol stress. The efforts allowed discovery of a mutant of slr0982 gene encoding an ATP-binding cassette transporter which grew poorly in BG11 medium supplemented with 1.5% (v/v ethanol when compared with the wild type, and the growth loss could be recovered by complementing slr0982 in the ∆slr0982 mutant, suggesting that slr0982 is involved in ethanol tolerance in Synechocystis. To decipher the tolerance mechanism involved, a comparative metabolomic and network-based analysis of the wild type and the ethanol-sensitive ∆slr0982 mutant was performed. The analysis allowed the identification of four metabolic modules related to slr0982 deletion in the ∆slr0982 mutant, among which metabolites like sucrose and L-pyroglutamic acid which might be involved in ethanol tolerance, were found important for slr0982 deletion in the ∆slr0982 mutant. This study reports on the first transporter related to ethanol tolerance in Synechocystis, which could be a useful target for further tolerance engineering. In addition, metabolomic and network analysis provides important findings for better understanding of the tolerance mechanism to ethanol stress in Synechocystis.

  12. Ethylene Regulates the Physiology of the Cyanobacterium Synechocystis sp. PCC 6803 via an Ethylene Receptor.

    Science.gov (United States)

    Lacey, Randy F; Binder, Brad M

    2016-08-01

    Ethylene is a plant hormone that plays a crucial role in the growth and development of plants. The ethylene receptors in plants are well studied, and it is generally assumed that they are found only in plants. In a search of sequenced genomes, we found that many bacterial species contain putative ethylene receptors. Plants acquired many proteins from cyanobacteria as a result of the endosymbiotic event that led to chloroplasts. We provide data that the cyanobacterium Synechocystis (Synechocystis sp. PCC 6803) has a functional receptor for ethylene, Synechocystis Ethylene Response1 (SynEtr1). We first show that SynEtr1 directly binds ethylene. Second, we demonstrate that application of ethylene to Synechocystis cells or disruption of the SynEtr1 gene affects several processes, including phototaxis, type IV pilus biosynthesis, photosystem II levels, biofilm formation, and spontaneous cell sedimentation. Our data suggest a model where SynEtr1 inhibits downstream signaling and ethylene inhibits SynEtr1. This is similar to the inverse-agonist model of ethylene receptor signaling proposed for plants and suggests a conservation of structure and function that possibly originated over 1 billion years ago. Prior research showed that SynEtr1 also contains a light-responsive phytochrome-like domain. Thus, SynEtr1 is a bifunctional receptor that mediates responses to both light and ethylene. To our knowledge, this is the first demonstration of a functional ethylene receptor in a nonplant species and suggests that that the perception of ethylene is more widespread than previously thought. © 2016 American Society of Plant Biologists. All Rights Reserved.

  13. [Increasing reductant NADPH content via metabolic engineering of PHB synthesis pathway in Synechocystis sp. PCC 6803].

    Science.gov (United States)

    Xie, Juan; Zhou, Jie; Zhang, Haifeng; Li, Yin

    2011-07-01

    Cyanobacteria have become attractive hosts for renewable chemicals production. The low productivity, however, prevents it from industrial application. Reductant NAD(P)H availability is a chief hurdle for the production of reductive metabolites in microbes. To increase NADPH content in Synechocystis sp. PCC 6803, PHB synthase encoding gene phaC and phaE in Synechocystis was inactivated by replacing phaC&E genes with chloromycetin resistance cassette via homologous recombination. PCR analysis showed that mutant S.delta phaC&E with complete genome segregation was generated. The comparison between growth curves of S.wt and S.delta phaC&E indicated the knockout of phaC & phaE genes did not affect obviously the cell growth. Gas chromatography analysis showed that the accumulation of PHB in wild type was about 2.3% of the dry cell weight, whereas no PHB was detected in the mutant S.delta phaC&E. The data indicated that inactivation of PHB synthase gene phaC and phaE interrupted the synthesis of PHB. Further comparative study of wild type and mutant demonstrated that NADPH content in S.delta phaC&E was obviously increased. On the third day, the NADPH content in S.delta phaC&E was up to 1.85 fold higher than that in wild type. These results indicated that deleting PHB synthase gene phaC and phaE not only can block the synthesis of PHB, but also can save NADPH to contribute reductant sink in cyanobacteria. Hence, the engineered cyanobacterial strain S.delta phaC&E, in which carbon flux was redirected and NADPH was increased, will be a potential host strain for chemicals production in cyanobacteria.

  14. Phosphoproteome of the cyanobacterium Synechocystis sp. PCC 6803 and its dynamics during nitrogen starvation.

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    Philipp eSpät

    2015-03-01

    Full Text Available Cyanobacteria have shaped the earth’s biosphere as the first oxygenic photoautotrophs and still play an important role in many ecosystems. The ability to adapt to changing environmental conditions is an essential characteristic in order to ensure survival. To this end, numerous studies have shown that bacteria use protein post-translational modifications such as Ser/Thr/Tyr phosphorylation in cell signalling, adaptation and regulation. Nevertheless, our knowledge of cyanobacterial phosphoproteomes and their dynamic response to environmental stimuli is relatively limited. In this study, we applied gel-free methods and high accuracy mass spectrometry towards the unbiased detection of Ser/Thr/Tyr phosphorylation events in the model cyanobacterium Synechocystis sp. PCC 6803. We could identify over 300 phosphorylation events in cultures grown on nitrate as exclusive nitrogen source. Chemical dimethylation labelling was applied to investigate proteome and phosphoproteome dynamics during nitrogen starvation. Our dataset describes the most comprehensive (phosphoproteome of Synechocystis to date, identifying 2,382 proteins and 183 phosphorylation events and quantifying 2,111 proteins and 148 phosphorylation events during nitrogen starvation. Global protein phosphorylation levels were increased in response to nitrogen depletion after 24 hours. Among the proteins with increased phosphorylation, the PII signalling protein showed the highest fold-change, serving as positive control. Other proteins with increased phosphorylation levels comprised functions in photosynthesis and in carbon and nitrogen metabolism. This study reveals dynamics of Synechocystis phosphoproteome in response to environmental stimuli and suggests an important role of protein Ser/Thr/Tyr phosphorylation in fundamental mechanisms of homeostatic control in cyanobacteria.

  15. The application of two-step linear temperature program to thermal analysis for monitoring the lipid induction of Nostoc sp. KNUA003 in large scale cultivation.

    Science.gov (United States)

    Kang, Bongmun; Yoon, Ho-Sung

    2015-02-01

    Recently, microalgae was considered as a renewable energy for fuel production because its production is nonseasonal and may take place on nonarable land. Despite all of these advantages, microalgal oil production is significantly affected by environmental factors. Furthermore, the large variability remains an important problem in measurement of algae productivity and compositional analysis, especially, the total lipid content. Thus, there is considerable interest in accurate determination of total lipid content during the biotechnological process. For these reason, various high-throughput technologies were suggested for accurate measurement of total lipids contained in the microorganisms, especially oleaginous microalgae. In addition, more advanced technologies were employed to quantify the total lipids of the microalgae without a pretreatment. However, these methods are difficult to measure total lipid content in wet form microalgae obtained from large-scale production. In present study, the thermal analysis performed with two-step linear temeperature program was applied to measure heat evolved in temperature range from 310 to 351 °C of Nostoc sp. KNUA003 obtained from large-scale cultivation. And then, we examined the relationship between the heat evolved in 310-351 °C (HE) and total lipid content of the wet Nostoc cell cultivated in raceway. As a result, the linear relationship was determined between HE value and total lipid content of Nostoc sp. KNUA003. Particularly, there was a linear relationship of 98% between the HE value and the total lipid content of the tested microorganism. Based on this relationship, the total lipid content converted from the heat evolved of wet Nostoc sp. KNUA003 could be used for monitoring its lipid induction in large-scale cultivation. Copyright © 2014 Elsevier Inc. All rights reserved.

  16. Engineering Limonene and Bisabolene Production in Wild Type and a Glycogen-Deficient Mutant of Synechococcus sp. PCC 7002

    Energy Technology Data Exchange (ETDEWEB)

    Davies, Fiona K., E-mail: fdavies@mines.edu [Department of Chemistry and Geochemistry, Colorado School of Mines, Golden, CO (United States); Work, Victoria H. [Civil and Environmental Engineering Division, Colorado School of Mines, Golden, CO (United States); Beliaev, Alexander S. [Biological Sciences Division, Pacific Northwest National Laboratory, Richland, WA (United States); Posewitz, Matthew C. [Department of Chemistry and Geochemistry, Colorado School of Mines, Golden, CO (United States)

    2014-06-19

    The plant terpenoids limonene (C{sub 10}H{sub 16}) and α-bisabolene (C{sub 15}H{sub 24}) are hydrocarbon precursors to a range of industrially relevant chemicals. High-titer microbial synthesis of limonene and α-bisabolene could pave the way for advances in in vivo engineering of tailor-made hydrocarbons, and production at commercial scale. We have engineered the fast-growing unicellular euryhaline cyanobacterium Synechococcus sp. PCC 7002 to produce yields of 4 mg L{sup −1} limonene and 0.6 mg L{sup −1} α-bisabolene through heterologous expression of the Mentha spicatal-limonene synthase or the Abies grandis (E)-α-bisabolene synthase genes, respectively. Titers were significantly higher when a dodecane overlay was applied during culturing, suggesting either that dodecane traps large quantities of volatile limonene or α-bisabolene that would otherwise be lost to evaporation, and/or that continuous product removal in dodecane alleviates product feedback inhibition to promote higher rates of synthesis. We also investigate limonene and bisabolene production in the ΔglgC genetic background, where carbon partitioning is redirected at the expense of glycogen biosynthesis. The Synechococcus sp. PCC 7002 ΔglgC mutant excreted a suite of overflow metabolites (α-ketoisocaproate, pyruvate, α-ketoglutarate, succinate, and acetate) during nitrogen-deprivation, and also at the onset of stationary growth in nutrient-replete media. None of the excreted metabolites, however, appeared to be effectively utilized for terpenoid metabolism. Interestingly, we observed a 1.6- to 2.5-fold increase in the extracellular concentration of most excreted organic acids when the ΔglgC mutant was conferred with the ability to produce limonene. Overall, Synechococcus sp. PCC 7002 provides a highly promising platform for terpenoid biosynthetic and metabolic engineering efforts.

  17. Engineering limonene and bisabolene production in wild type and a glycogen-deficient mutant of Synechococcus sp. PCC 7002

    Energy Technology Data Exchange (ETDEWEB)

    Davies, Fiona K.; Work, Victoria H.; Beliaev, Alex S.; Posewitz, Matthew C.

    2014-06-19

    The plant terpenoids limonene (C10H16) and α-bisabolene (C15H24) are hydrocarbon precursors to a range of industrially-relevant chemicals. High-titer microbial synthesis of limonene and α- bisabolene could pave the way for advances in in vivo engineering of tailor-made hydrocarbons, and production at commercial scale. We have engineered the fast-growing unicellular euryhaline cyanobacterium Synechococcus sp. PCC 7002 to produce yields of 4 mg L-1 limonene and 0.6 mg L-1 α-bisabolene through heterologous expression of the Mentha spicata L-limonene synthase or the Abies grandis (E)-α-bisabolene synthase genes, respectively. Titers were significantly higher when a dodecane overlay was applied during culturing, suggesting either that dodecane traps large quantities of volatile limonene and α-bisabolene that would otherwise be lost to evaporation, and/or that continuous product removal in dodecane alleviates product feedback inhibition to promote higher rates of synthesis. We also investigate limonene and bisabolene production in the ΔglgC genetic background, where carbon partitioning is redirected at the expense of glycogen biosynthesis. The Synechococcus sp. PCC 7002 ΔglgC mutant excreted a suite of overflow metabolites (α-ketoisocaproate, pyruvate, α-ketoglutarate, succinate and acetate) during nitrogen deprivation, and also at the onset of stationary growth in nutrient-replete media. None of the excreted metabolites, however, appeared to be effectively utilized for terpenoid metabolism. Interestingly, we observed a 1.6 to 2.5-fold increase in the extracellular concentration of most excreted organic acids when the ΔglgC mutant was conferred with the ability to produce limonene. Overall, Synechococcus sp. PCC 7002 provides a highly promising platform for terpenoid biosynthetic and metabolic engineering efforts.

  18. Engineering Limonene and Bisabolene Production in Wild Type and a Glycogen-Deficient Mutant of Synechococcus sp. PCC 7002.

    Science.gov (United States)

    Davies, Fiona K; Work, Victoria H; Beliaev, Alexander S; Posewitz, Matthew C

    2014-01-01

    The plant terpenoids limonene (C10H16) and α-bisabolene (C15H24) are hydrocarbon precursors to a range of industrially relevant chemicals. High-titer microbial synthesis of limonene and α-bisabolene could pave the way for advances in in vivo engineering of tailor-made hydrocarbons, and production at commercial scale. We have engineered the fast-growing unicellular euryhaline cyanobacterium Synechococcus sp. PCC 7002 to produce yields of 4 mg L(-1) limonene and 0.6 mg L(-1) α-bisabolene through heterologous expression of the Mentha spicatal-limonene synthase or the Abies grandis (E)-α-bisabolene synthase genes, respectively. Titers were significantly higher when a dodecane overlay was applied during culturing, suggesting either that dodecane traps large quantities of volatile limonene or α-bisabolene that would otherwise be lost to evaporation, and/or that continuous product removal in dodecane alleviates product feedback inhibition to promote higher rates of synthesis. We also investigate limonene and bisabolene production in the ΔglgC genetic background, where carbon partitioning is redirected at the expense of glycogen biosynthesis. The Synechococcus sp. PCC 7002 ΔglgC mutant excreted a suite of overflow metabolites (α-ketoisocaproate, pyruvate, α-ketoglutarate, succinate, and acetate) during nitrogen-deprivation, and also at the onset of stationary growth in nutrient-replete media. None of the excreted metabolites, however, appeared to be effectively utilized for terpenoid metabolism. Interestingly, we observed a 1.6- to 2.5-fold increase in the extracellular concentration of most excreted organic acids when the ΔglgC mutant was conferred with the ability to produce limonene. Overall, Synechococcus sp. PCC 7002 provides a highly promising platform for terpenoid biosynthetic and metabolic engineering efforts.

  19. Engineering Limonene and Bisabolene Production in Wild Type and a Glycogen-Deficient Mutant of Synechococcus sp. PCC 7002

    International Nuclear Information System (INIS)

    Davies, Fiona K.; Work, Victoria H.; Beliaev, Alexander S.; Posewitz, Matthew C.

    2014-01-01

    The plant terpenoids limonene (C 10 H 16 ) and α-bisabolene (C 15 H 24 ) are hydrocarbon precursors to a range of industrially relevant chemicals. High-titer microbial synthesis of limonene and α-bisabolene could pave the way for advances in in vivo engineering of tailor-made hydrocarbons, and production at commercial scale. We have engineered the fast-growing unicellular euryhaline cyanobacterium Synechococcus sp. PCC 7002 to produce yields of 4 mg L −1 limonene and 0.6 mg L −1 α-bisabolene through heterologous expression of the Mentha spicatal-limonene synthase or the Abies grandis (E)-α-bisabolene synthase genes, respectively. Titers were significantly higher when a dodecane overlay was applied during culturing, suggesting either that dodecane traps large quantities of volatile limonene or α-bisabolene that would otherwise be lost to evaporation, and/or that continuous product removal in dodecane alleviates product feedback inhibition to promote higher rates of synthesis. We also investigate limonene and bisabolene production in the ΔglgC genetic background, where carbon partitioning is redirected at the expense of glycogen biosynthesis. The Synechococcus sp. PCC 7002 ΔglgC mutant excreted a suite of overflow metabolites (α-ketoisocaproate, pyruvate, α-ketoglutarate, succinate, and acetate) during nitrogen-deprivation, and also at the onset of stationary growth in nutrient-replete media. None of the excreted metabolites, however, appeared to be effectively utilized for terpenoid metabolism. Interestingly, we observed a 1.6- to 2.5-fold increase in the extracellular concentration of most excreted organic acids when the ΔglgC mutant was conferred with the ability to produce limonene. Overall, Synechococcus sp. PCC 7002 provides a highly promising platform for terpenoid biosynthetic and metabolic engineering efforts.

  20. Diurnal Regulation of Cellular Processes in the Cyanobacterium Synechocystis sp. Strain PCC 6803: Insights from Transcriptomic, Fluxomic, and Physiological Analyses

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    Rajib Saha

    2016-05-01

    Full Text Available Synechocystis sp. strain PCC 6803 is the most widely studied model cyanobacterium, with a well-developed omics level knowledgebase. Like the lifestyles of other cyanobacteria, that of Synechocystis PCC 6803 is tuned to diurnal changes in light intensity. In this study, we analyzed the expression patterns of all of the genes of this cyanobacterium over two consecutive diurnal periods. Using stringent criteria, we determined that the transcript levels of nearly 40% of the genes in Synechocystis PCC 6803 show robust diurnal oscillating behavior, with a majority of the transcripts being upregulated during the early light period. Such transcripts corresponded to a wide array of cellular processes, such as light harvesting, photosynthetic light and dark reactions, and central carbon metabolism. In contrast, transcripts of membrane transporters for transition metals involved in the photosynthetic electron transport chain (e.g., iron, manganese, and copper were significantly upregulated during the late dark period. Thus, the pattern of global gene expression led to the development of two distinct transcriptional networks of coregulated oscillatory genes. These networks help describe how Synechocystis PCC 6803 regulates its metabolism toward the end of the dark period in anticipation of efficient photosynthesis during the early light period. Furthermore, in silico flux prediction of important cellular processes and experimental measurements of cellular ATP, NADP(H, and glycogen levels showed how this diurnal behavior influences its metabolic characteristics. In particular, NADPH/NADP+ showed a strong correlation with the majority of the genes whose expression peaks in the light. We conclude that this ratio is a key endogenous determinant of the diurnal behavior of this cyanobacterium.

  1. Involvement of sulfoquinovosyl diacylglycerol in DNA synthesis in Synechocystis sp. PCC 6803

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    Aoki Motohide

    2012-02-01

    Full Text Available Abstract Background Sulfoquinovosyl diacylglycerol (SQDG is present in the membranes of cyanobacteria and their postulated progeny, plastids, in plants. A cyanobacterium, Synechocystis sp. PCC 6803, requires SQDG for growth: its mutant (SD1 with the sqdB gene for SQDG synthesis disrupted can grow with external supplementation of SQDG. However, upon removal of SQDG from the medium, its growth is retarded, with a decrease in the cellular content of SQDG throughout cell division, and finally ceases. Concomitantly with the decrease in SQDG, the maximal activity of photosynthesis at high-light intensity is repressed by 40%. Findings We investigated effects of SQDG-defect on physiological aspects in Synechocystis with the use of SD1. SD1 cells defective in SQDG exhibited normal photosynthesis at low-light intensity as on culturing. Meanwhile, SD1 cells defective in SQDG were impaired in light-activated heterotrophic growth as well as in photoautotrophic growth. Flow cytometric analysis of the photoautotrophically growing cells gave similar cell size histograms for the wild type and SD1 supplemented with SQDG. However, the profile of SD1 defective in SQDG changed such that large part of the cell population was increased in size. Of particular interest was the microscopic observation that the mitotic index, i.e., population of dumbbell-like cells with a septum, increased from 14 to 29% in the SD1 culture without SQDG. Flow cytometric analysis also showed that the enlarged cells of SD1 defective in SQDG contained high levels of Chl, however, the DNA content was low. Conclusions Our experiments strongly support the idea that photosynthesis is not the limiting factor for the growth of SD1 defective in SQDG, and that SQDG is responsible for some physiologically fundamental process common to both photoautotrophic and light-activated heterotrophic growth. Our findings suggest that the SQDG-defect allows construction of the photosynthetic machinery at an

  2. Gene expression patterns of sulfur starvation in Synechocystis sp. PCC 6803

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    Pendse Ninad D

    2008-07-01

    Full Text Available Abstract Background The unicellular cyanobacterium Synechocystis sp. PCC 6803 is a model microbe for studying biochemistry, genetics and molecular biology of photobiological processes. Importance of this bacterium in basic and applied research calls for a systematic, genome-wide description of its transcriptional regulatory capacity. Characteristic transcriptional responses to changes in the growth environment are expected to provide a scaffold for describing the Synechocystis transcriptional regulatory network as well as efficient means for functional annotation of genes in the genome. Results We designed, validated and used Synechocystis genome-wide oligonucleotide (70-mer microarray (representing 96.7% of all chromosomal ORFs annotated at the time of the beginning of this project to study transcriptional activity of the cyanobacterial genome in response to sulfur (S starvation. The microarray data were verified by quantitative RT-PCR. We made five main observations: 1 Transcriptional changes upon sulfate starvation were relatively moderate, but significant and consistent with growth kinetics; 2 S acquisition genes encoding for a high-affinity sulfate transporter were significantly induced, while decreased transcription of genes for phycobilisome, photosystems I and II, cytochrome b6/f, and ATP synthase indicated reduced light-harvesting and photosynthetic activity; 3 S starvation elicited transcriptional responses associated with general growth arrest and stress; 4 A large number of genes regulated by S availability encode hypothetical proteins or proteins of unknown function; 5 Hydrogenase structural and maturation accessory genes were not identified as differentially expressed, even though increased hydrogen evolution was observed. Conclusion The expression profiles recorded by using this oligonucleotide-based microarray platform revealed that during transition from the condition of plentiful S to S starvation, Synechocystis undergoes

  3. Glutaredoxins are essential for stress adaptation in the cyanobacterium Synechocystis sp. PCC 6803.

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    Ana M Sánchez-Riego

    2013-11-01

    Full Text Available Glutaredoxin are small redox proteins able to reduce disulfides and mixed disulfides between GSH and proteins. Synechocystis sp. PCC 6803 contains three genes coding for glutaredoxins: ssr2061 (grxA and slr1562 (grxB code for dithiolic glutaredoxins while slr1846 (grxC codes for a monothiolic glutaredoxin. We have analyzed the expression of these glutaredoxins in response to different stresses, such as high light, H2O2 and heat shock. Analysis of the mRNA levels showed that grxA is only induced by heat while grxC is repressed by heat shock and is induced by high light and H2O2. In contrast, grxB expression was maintained almost constant under all conditions. Analysis of GrxA and GrxC protein levels by western blot showed that GrxA increases in response to high light, heat or H2O2 while GrxC is only induced by high light and H2O2, in accordance with its mRNA levels. In addition, we have also generated mutants that have interrupted one, two or three glutaredoxin genes. These mutants were viable and did not show any different phenotype from the WT under standard growth conditions. Nevertheless, analysis of these mutants under several stress conditions revealed that single grxA mutants grow slower after H2O2, heat and high light treatments, while mutants in grxB are indistinguishable from WT. grxC mutants were hypersensitive to treatments with H2O2, heat, high light and metals. A double grxAgrxC mutant was found to be even more sensitive to H2O2 than each corresponding single mutants. Surprisingly a mutation in grxB suppressed totally or partially the phenotypes of grxA and grxC mutants except the H2O2 sensitivity of the grxC mutant. This suggests that grxA and grxC participate in independent pathways while grxA and grxB participate in a common pathway for H2O2 resistance. The data presented here show that glutaredoxins are essential for stress adaptation in cyanobacteria, although their targets and mechanism of action remain unidentified.

  4. Photoautotrophic Production of Biomass, Laurate, and Soluble Organics by Synechocystis sp. PCC 6803

    Science.gov (United States)

    Nguyen, Binh Thanh

    Photosynthesis converts sunlight to biomass at a global scale. Among the photosynthetic organisms, cyanobacteria provide an excellent model to study how photosynthesis can become a practical platform of large-scale biotechnology. One novel approach involves metabolically engineering the cyanobacterium Synechocystis sp. PCC 6803 to excrete laurate, which is harvested directly. This work begins by defining a working window of light intensity (LI). Wild-type and laurate-excreting Synechocystis required an LI of at least 5 muE/m2-s to sustain themselves, but are photo-inhibited by LI of 346 to 598 muE/m2-s. Fixing electrons into valuable organic products, e.g., biomass and excreted laurate, is critical to success. Wild-type Synechocystis channeled 75% to 84% of its fixed electrons to biomass; laurate-excreting Synechocystis fixed 64 to 69% as biomass and 6.6% to 10% as laurate. This means that 16 to 30% of the electrons were diverted to non-valuable soluble products, and the trend was accentuated with higher LI. How the Ci concentration depended on the pH and the nitrogen source was quantified by the proton condition and experimentally validated. Nitrate increased, ammonium decreased, but ammonium nitrate stabilized alkalinity and Ci. This finding provides a mechanistically sound tool to manage Ci and pH independently. Independent evaluation pH and Ci on the growth kinetics of Synechocystis showed that pH 8.5 supported the fastest maximum specific growth rate (mumax): 2.4/day and 1.7/day, respectively, for the wild type and modified strains with LI of 202 muE/m2-s. Half-maximum-rate concentrations (KCi) were less than 0.1 mM, meaning that Synechocystis should attain its mumax with a modest Ci concentration (≥1.0 mM). Biomass grown with day-night cycles had a night endogenous decay rate of 0.05-1.0/day, with decay being faster with higher LI and the beginning of dark periods. Supplying light at a fraction of daylight reduced dark decay rate and improved overall

  5. Directional RNA deep sequencing sheds new light on the transcriptional response of Anabaena sp. strain PCC 7120 to combined-nitrogen deprivation

    Directory of Open Access Journals (Sweden)

    Head Steven R

    2011-06-01

    Full Text Available Abstract Background Cyanobacteria are potential sources of renewable chemicals and biofuels and serve as model organisms for bacterial photosynthesis, nitrogen fixation, and responses to environmental changes. Anabaena (Nostoc sp. strain PCC 7120 (hereafter Anabaena is a multicellular filamentous cyanobacterium that can "fix" atmospheric nitrogen into ammonia when grown in the absence of a source of combined nitrogen. Because the nitrogenase enzyme is oxygen sensitive, Anabaena forms specialized cells called heterocysts that create a microoxic environment for nitrogen fixation. We have employed directional RNA-seq to map the Anabaena transcriptome during vegetative cell growth and in response to combined-nitrogen deprivation, which induces filaments to undergo heterocyst development. Our data provide an unprecedented view of transcriptional changes in Anabaena filaments during the induction of heterocyst development and transition to diazotrophic growth. Results Using the Illumina short read platform and a directional RNA-seq protocol, we obtained deep sequencing data for RNA extracted from filaments at 0, 6, 12, and 21 hours after the removal of combined nitrogen. The RNA-seq data provided information on transcript abundance and boundaries for the entire transcriptome. From these data, we detected novel antisense transcripts within the UTRs (untranslated regions and coding regions of key genes involved in heterocyst development, suggesting that antisense RNAs may be important regulators of the nitrogen response. In addition, many 5' UTRs were longer than anticipated, sometimes extending into upstream open reading frames (ORFs, and operons often showed complex structure and regulation. Finally, many genes that had not been previously identified as being involved in heterocyst development showed regulation, providing new candidates for future studies in this model organism. Conclusions Directional RNA-seq data were obtained that provide

  6. Enzyme kinetics, inhibitors, mutagenesis and electron paramagnetic resonance analysis of dual-affinity nitrate reductase in unicellular N(2)-fixing cyanobacterium Cyanothece sp. PCC 8801.

    Science.gov (United States)

    Wang, Tung-Hei; Chen, Yung-Han; Huang, Jine-Yung; Liu, Kang-Cheng; Ke, Shyue-Chu; Chu, Hsiu-An

    2011-11-01

    The assimilatory nitrate reductase (NarB) of N(2)-fixing cyanobacterium Cyanothece sp. PCC 8801 is a monomeric enzyme with dual affinity for substrate nitrate. We purified the recombinant NarB of Cyanothece sp. PCC 8801 and further investigated it by enzyme kinetics analysis, site-directed mutagenesis, inhibitor kinetics analysis, and electron paramagnetic resonance (EPR) spectroscopy. The NarB showed 2 kinetic regimes at pH 10.5 or 8 and electron-donor conditions methyl viologen or ferredoxin (Fd). Fd-dependent NR assay revealed NarB with very high affinity for nitrate (K(m)1, ∼1μM; K(m)2, ∼270μM). Metal analysis and EPR results showed that NarB contains a Mo cofactor and a [4Fe-4S] cluster. In addition, the R352A mutation on the proposed nitrate-binding site of NarB greatly altered both high- and low-affinity kinetic components. Furthermore, the effect of azide on the NarB of Cyanothece sp. PCC 8801 was more complex than that on the NarB of Synechococcus sp. PCC 7942 with its single kinetic regime. With 1mM azide, the kinetics of the wild-type NarB was transformed from 2 kinetic regimes to hyperbolic kinetics, and its activity was enhanced significantly under medium nitrate concentrations. Moreover, EPR results also suggested a structural difference between the two NarBs. Taken together, our results show that the NarB of Cyanothece sp. PCC 8801 contains only a single Mo-catalytic center, and we rule out that the enzyme has 2 independent, distinct catalytic sites. In addition, the NarB of Cyanothece sp. PCC 8801 may have a regulatory nitrate-binding site. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

  7. MapA, an iron-regulated, cytoplasmic membrane protein in the cyanobacterium Synechococcus sp. strain PCC7942.

    Science.gov (United States)

    Webb, R; Troyan, T; Sherman, D; Sherman, L A

    1994-08-01

    Growth of Synechococcus sp. strain PCC 7942 in iron-deficient media leads to the accumulation of an approximately 34-kDa protein. The gene encoding this protein, mapA (membrane-associated protein A), has been cloned and sequenced (GenBank accession number, L01621). The mapA transcript is not detectable in normally grown cultures but is stably accumulated by cells grown in iron-deficient media. However, the promoter sequence for this gene does not resemble other bacterial iron-regulated promoters described to date. The carboxyl-terminal region of the derived amino acid sequence of MapA resembles bacterial proteins involved in iron acquisition, whereas the amino-terminal end of MapA has a high degree of amino acid identity with the abundant, chloroplast envelope protein E37. An approach employing improved cellular fractionation techniques as well as electron microscopy and immunocytochemistry was essential in localizing MapA protein to the cytoplasmic membrane of Synechococcus sp. strain PCC 7942. When these cells were grown under iron-deficient conditions, a significant fraction of MapA could also be localized to the thylakoid membranes.

  8. Growth of Chlamydomonas reinhardtii in acetate-free medium when co-cultured with alginate-encapsulated, acetate-producing strains of Synechococcus sp. PCC 7002.

    Science.gov (United States)

    Therien, Jesse B; Zadvornyy, Oleg A; Posewitz, Matthew C; Bryant, Donald A; Peters, John W

    2014-01-01

    The model alga Chlamydomonas reinhardtii requires acetate as a co-substrate for optimal production of lipids, and the addition of acetate to culture media has practical and economic implications for algal biofuel production. Here we demonstrate the growth of C. reinhardtii on acetate provided by mutant strains of the cyanobacterium Synechococcus sp. PCC 7002. Optimal growth conditions for co-cultivation of C. reinhardtii with wild-type and mutant strains of Synechococcus sp. 7002 were established. In co-culture, acetate produced by a glycogen synthase knockout mutant of Synechococcus sp. PCC 7002 was able to support the growth of a lipid-accumulating mutant strain of C. reinhardtii defective in starch production. Encapsulation of Synechococcus sp. PCC 7002 using an alginate matrix was successfully employed in co-cultures to limit growth and maintain the stability. The ability of immobilized strains of the cyanobacterium Synechococcus sp. PCC 7002 to produce acetate at a level adequate to support the growth of lipid-accumulating strains of C. reinhartdii offers a potentially practical, photosynthetic alternative to providing exogenous acetate into growth media.

  9. Light influences cytokinin biosynthesis and sensing in Nostoc (cyanobacteria).

    Science.gov (United States)

    Frébortová, Jitka; Plíhal, Ondřej; Florová, Vendula; Kokáš, Filip; Kubiasová, Karolina; Greplová, Marta; Šimura, Jan; Novák, Ondřej; Frébort, Ivo

    2017-06-01

    Cytokinins are an important group of plant hormones that are also found in other organisms, including cyanobacteria. While various aspects of cytokinin function and metabolism are well understood in plants, the information is limited for cyanobacteria. In this study, we first experimentally confirmed a prenylation of tRNA by recombinant isopentenyl transferase NoIPT2 from Nostoc sp. PCC 7120, whose encoding gene we previously identified in Nostoc genome along with the gene for adenylate isopentenyl transferase NoIPT1. In contrast to NoIPT2, the transcription of NoIPT1 was strongly activated during the dark period and was followed by an increase in the cytokinin content several hours later in the light period. Dominant cytokinin metabolites detected at all time points were free bases and monophosphates of isopentenyladenine and cis-zeatin, while N-glucosides were not detected at all. Whole transcriptome differential expression analysis of cultures of the above Nostoc strain treated by cytokinin compared to untreated controls indicated that cytokinin together with light trigger expression of several genes related to signal transduction, including two-component sensor histidine kinases and two-component hybrid sensors and regulators. One of the affected histidine kinases with a cyclase/histidine kinase-associated sensory extracellular domain similar to the cytokinin-binding domain in plant cytokinin receptors was able to modestly bind isopentenyladenine. The data show that the genetic disposition allows Nostoc not only to produce free cytokinins and prenylate tRNA but also modulate the cytokinin biosynthesis in response to light, triggering complex changes in sensing and regulation. © 2017 Phycological Society of America.

  10. Concerted changes in gene expression and cell physiology of the cyanobacterium Synechocystis sp. strain PCC 6803 during transitions between nitrogen and light-limited growth

    NARCIS (Netherlands)

    Aquirre von Wobeser, E.; Ibelings, B.W.; Bok, J.M.; Krasikov, V.; Huisman, J.; Matthijs, H.C.P.

    2011-01-01

    Physiological adaptation and genome-wide expression profiles of the cyanobacterium Synechocystis sp. strain PCC 6803 in response to gradual transitions between nitrogen-limited and light-limited growth conditions were measured in continuous cultures. Transitions induced changes in pigment

  11. The Genome Sequence of the Cyanobacterium Oscillatoria sp. PCC 6506 Reveals Several Gene Clusters Responsible for the Biosynthesis of Toxins and Secondary Metabolites▿

    Science.gov (United States)

    Méjean, Annick; Mazmouz, Rabia; Mann, Stéphane; Calteau, Alexandra; Médigue, Claudine; Ploux, Olivier

    2010-01-01

    We report a draft sequence of the genome of Oscillatoria sp. PCC 6506, a cyanobacterium that produces anatoxin-a and homoanatoxin-a, two neurotoxins, and cylindrospermopsin, a cytotoxin. Beside the clusters of genes responsible for the biosynthesis of these toxins, we have found other clusters of genes likely involved in the biosynthesis of not-yet-identified secondary metabolites. PMID:20675499

  12. The role of Slr0151, a tetratricopeptide repeat protein from Synechocystis sp. PCC 6803, during Photosystem II assembly and repair

    Directory of Open Access Journals (Sweden)

    Anna eRast

    2016-05-01

    Full Text Available The assembly and repair of photosystem II (PSII is facilitated by a variety of assembly factors. Among those, the tetratricopeptide repeat (TPR protein Slr0151 from Synechocystis sp. PCC 6803 (hereafter Synechocystis has previously been assigned a repair function under high light conditions (Yang et al., 2014, J. Integr. Plant Biol. 56, 1136-50. Here, we show that inactivation of Slr0151 affects thylakoid membrane ultrastructure even under normal light conditions. Moreover, the level and localization of Slr0151 are affected in a variety of PSII-related mutants. In particular, the data suggest a close functional relationship between Slr0151 and Sll0933, which interacts with Ycf48 during PSII assembly and is homologous to PAM68 in Arabidopsis thaliana. Immunofluorescence analysis revealed a punctate distribution of Slr0151 within several different membrane types in Synechocystis cells.

  13. Responses of Synechocystis sp. PCC 6803 to heterologous biosynthetic pathways

    DEFF Research Database (Denmark)

    Vavitsas, Konstantinos; Rue, Emil Østergaard; Stefánsdóttir, Lára Kristín

    2017-01-01

    BACKGROUND: There are an increasing number of studies regarding genetic manipulation of cyanobacteria to produce commercially interesting compounds. The majority of these works study the expression and optimization of a selected heterologous pathway, largely ignoring the wholeness and complexity...... different compounds, the cyanogenic glucoside dhurrin and the diterpenoid 13R-manoyl oxide in Synechocystis PCC 6803. We used genome-scale metabolic modelling to study fluxes in individual reactions and pathways, and we determined the concentrations of key metabolites, such as amino acids, carotenoids...

  14. Higher production of C-phycocyanin by nitrogen-free (diazotrophic) cultivation of Nostoc sp. NK and simplified extraction by dark-cold shock.

    Science.gov (United States)

    Lee, Na Kyeong; Oh, Hee-Mock; Kim, Hee-Sik; Ahn, Chi-Yong

    2017-03-01

    Nostoc sp. NK (KCTC 12772BP) was isolated and cultivated in a BG11 medium and a nitrate-free BG11 medium (BG11 0 ). To enhance C-phycocyanin (C-PC) content in the cells, different fluorescent lamps (white, plant, and red) were used as light sources for complementary chromatic adaptation (CCA). The maximum biomass productivity was 0.42g/L/d and 0.32g/L/d under BG11 and BG11 0 conditions, respectively. The maximum C-PC contents were 8.4% (w/w) under white lamps, 13.6% (w/w) under plant lamps, and 18% (w/w) under BG11 0 and the red light condition. The maximum C-PC productivity was 57.4mg/L/d in BG11 0 under the red lamp condition. These results indicate that a higher C-PC content could be obtained under a diazotrophic condition and a CCA reaction. The C-PC could be released naturally from cells without any extraction processes, when Nostoc sp. NK was cultivated in the BG11 0 medium with CO 2 aeration and put in dark conditions at 5°C. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. Quantitative proteomics reveals dynamic responses of Synechocystis sp. PCC 6803 to next-generation biofuel butanol.

    Science.gov (United States)

    Tian, Xiaoxu; Chen, Lei; Wang, Jiangxin; Qiao, Jianjun; Zhang, Weiwen

    2013-01-14

    Butanol is a promising biofuel, and recent metabolic engineering efforts have demonstrated the use of photosynthetic cyanobacterial hosts for its production. However, cyanobacteria have very low tolerance to butanol, limiting the economic viability of butanol production from these renewable producing systems. The existing knowledge of molecular mechanism involved in butanol tolerance in cyanobacteria is very limited. To build a foundation necessary to engineer robust butanol-producing cyanobacterial hosts, in this study, the responses of Synechocystis PCC 6803 to butanol were investigated using a quantitative proteomics approach with iTRAQ - LC-MS/MS technologies. The resulting high-quality dataset consisted of 25,347 peptides corresponding to 1452 unique proteins, a coverage of approximately 40% of the predicted proteins in Synechocystis. Comparative quantification of protein abundances led to the identification of 303 differentially regulated proteins by butanol. Annotation and GO term enrichment analysis showed that multiple biological processes were regulated, suggesting that Synechocystis probably employed multiple and synergistic resistance mechanisms in dealing with butanol stress. Notably, the analysis revealed the induction of heat-shock protein and transporters, along with modification of cell membrane and envelope were the major protection mechanisms against butanol. A conceptual cellular model of Synechocystis PCC 6803 responses to butanol stress was constructed to illustrate the putative molecular mechanisms employed to defend against butanol stress. Copyright © 2012 Elsevier B.V. All rights reserved.

  16. Two-Step Separation of Nostotrebin 6 from Cultivated Soil Cyanobacterium (Nostoc sp.) by High Performance Countercurrent Chromatography

    Czech Academy of Sciences Publication Activity Database

    Cheel, José; Kučerová, P.; Garrard, I.; Ignatova, S.; Hrouzek, Pavel; Kopecký, Jiří

    2014-01-01

    Roč. 19, č. 4 (2014), s. 8773-8787 ISSN 1420-3049 R&D Projects: GA MŠk ED2.1.00/03.0110; GA MŠk EE2.3.30.0059 Institutional support: RVO:61388971 Keywords : nostotrebin 6 * cyanobacterium * Nostoc * HPLC separation Subject RIV: EE - Microbiology, Virology Impact factor: 2.416, year: 2014

  17. Photosynthetic Versatility in the Genome of Geitlerinema sp. PCC 9228 (Formerly Oscillatoria limnetica 'Solar Lake'), a Model Anoxygenic Photosynthetic Cyanobacterium.

    Science.gov (United States)

    Grim, Sharon L; Dick, Gregory J

    2016-01-01

    Anoxygenic cyanobacteria that use sulfide as the electron donor for photosynthesis are a potentially influential but poorly constrained force on Earth's biogeochemistry. Their versatile metabolism may have boosted primary production and nitrogen cycling in euxinic coastal margins in the Proterozoic. In addition, they represent a biological mechanism for limiting the accumulation of atmospheric oxygen, especially before the Great Oxidation Event and in the low-oxygen conditions of the Proterozoic. In this study, we describe the draft genome sequence of Geitlerinema sp. PCC 9228, formerly Oscillatoria limnetica 'Solar Lake', a mat-forming diazotrophic cyanobacterium that can switch between oxygenic photosynthesis and sulfide-based anoxygenic photosynthesis (AP). Geitlerinema possesses three variants of psbA , which encodes protein D1, a core component of the photosystem II reaction center. Phylogenetic analyses indicate that one variant is closely affiliated with cyanobacterial psbA genes that code for a D1 protein used for oxygen-sensitive processes. Another version is phylogenetically similar to cyanobacterial psbA genes that encode D1 proteins used under microaerobic conditions, and the third variant may be cued to high light and/or elevated oxygen concentrations. Geitlerinema has the canonical gene for sulfide quinone reductase (SQR) used in cyanobacterial AP and a putative transcriptional regulatory gene in the same operon. Another operon with a second, distinct sqr and regulatory gene is present, and is phylogenetically related to sqr genes used for high sulfide concentrations. The genome has a comprehensive nif gene suite for nitrogen fixation, supporting previous observations of nitrogenase activity. Geitlerinema possesses a bidirectional hydrogenase rather than the uptake hydrogenase typically used by cyanobacteria in diazotrophy. Overall, the genome sequence of Geitlerinema sp. PCC 9228 highlights potential cyanobacterial strategies to cope with fluctuating

  18. МОРФОЛОГИЧЕСКАЯ И МОЛЕКУЛЯРНАЯ ХАРАТЕРИСТИКА ЦИАНОБАКТЕРИИ NOSTOC SP., ИЗОЛИРОВАННОЙ ИЗ ПОЧВЫ МУРМАНСКОЙ ОБЛАСТИ

    OpenAIRE

    ШАЛЫГИНА Р.Р.; ШАЛЫГИН С.С.; РЕДЬКИНА В.В.

    2016-01-01

    Штамм Nostoc sp., выделенный в чистую культуру из почв Кольского полуострова в районе Кандалакшского алюминиевого завода (КАЗ), отличается намного большим размером гетероцит и акинет по сравнению с другими видами рода Nostoc. Филогенетический анализ участка 16S рРНК показал, что данный штамм располагается точно в середине Nostoc sensu lato в кладе с Nostoc Bashkir 6A и Nostoc PCC9709. Мы предполагаем, что выделенный штамм является новым видом рода Nostoc sp., но для точного определения видово...

  19. Proteomic analysis reveals resistance mechanism against biofuel hexane in Synechocystis sp. PCC 6803

    Directory of Open Access Journals (Sweden)

    Liu Jie

    2012-09-01

    Full Text Available Abstract Background Recent studies have demonstrated that photosynthetic cyanobacteria could be an excellent cell factory to produce renewable biofuels and chemicals due to their capability to utilize solar energy and CO2 as the sole energy and carbon sources. Biosynthesis of carbon-neutral biofuel alkanes with good chemical and physical properties has been proposed. However, to make the process economically feasible, one major hurdle to improve the low cell tolerance to alkanes needed to be overcome. Results Towards the goal to develop robust and high-alkane-tolerant hosts, in this study, the responses of model cyanobacterial Synechocystis PCC 6803 to hexane, a representative of alkane, were investigated using a quantitative proteomics approach with iTRAQ - LC-MS/MS technologies. In total, 1,492 unique proteins were identified, representing about 42% of all predicted protein in the Synechocystis genome. Among all proteins identified, a total of 164 and 77 proteins were found up- and down-regulated, respectively. Functional annotation and KEGG pathway enrichment analyses showed that common stress responses were induced by hexane in Synechocystis. Notably, a large number of transporters and membrane-bound proteins, proteins against oxidative stress and proteins related to sulfur relay system and photosynthesis were induced, suggesting that they are possibly the major protection mechanisms against hexane toxicity. Conclusion The study provided the first comprehensive view of the complicated molecular mechanism employed by cyanobacterial model species, Synechocystis to defend against hexane stress. The study also provided a list of potential targets to engineer Synechocystis against hexane stress.

  20. Impacts of diurnal variation of ultraviolet-B and photosynthetically active radiation on phycobiliproteins of the hot-spring cyanobacterium Nostoc sp. strain HKAR-2.

    Science.gov (United States)

    Kannaujiya, Vinod K; Sinha, Rajeshwar P

    2017-01-01

    The effects of diurnal variation of photosynthetically active radiation (PAR; 400-700 nm) and ultraviolet-B (UV-B; 280-315 nm) radiation on phycobiliproteins (PBPs) and photosynthetic pigments (PP) have been studied in the hot-spring cyanobacterium Nostoc sp. strain HKAR-2. The variations in PBPs and PP were monitored by alternating light and dark under PAR, UV-B, and PAR + UV-B radiations over a period of 25 h. There was a decline in the amount of Chl a and PBPs during light periods of UV-B and PAR + UV-B and an increase during dark periods showing a circadian rhythm by destruction and resynthesis of pigment-protein complex. However, a marked induction in carotenoids was recorded during light periods of the same radiations. Moreover, the ratio of Chl a/PE and Chl a/PC was increased in dark periods showing the resynthesis of bleached Chl a. The wavelength shift in emission fluorescence of PBPs toward shorter wavelengths further indicated the bleaching and destruction of PBPs during light periods. Oxidative damage upon exposure to PAR, UV-B, and PAR + UV-B was alleviated by induction of antioxidative enzymes such as superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX). The studied cyanobacterium exhibits a significant increase in the activities of SOD, CAT, and APX upon exposure to UV-B and PAR + UV-B radiations. The results indicate that pigment-protein composition of Nostoc sp. stain HKAR-2 was significantly altered during diurnal variation of light/radiation, which might play an important role in optimization for their productivity in a particular cyanobacterium.

  1. Increasing the Photoautotrophic Growth Rate of Synechocystis sp. PCC 6803 by Identifying the Limitations of Its Cultivation.

    Science.gov (United States)

    van Alphen, Pascal; Abedini Najafabadi, Hamed; Branco Dos Santos, Filipe; Hellingwerf, Klaas J

    2018-03-25

    Many conditions have to be optimized in order to be able to grow the cyanobacterium Synechocystis sp. PCC 6803 (Synechocystis) for an extended period of time under physiologically well-defined and constant conditions. It is still poorly understood what limits growth of this organism in batch and continuous cultures in BG-11, the standard medium used to grow Synechocystis. Through a series of batch experiments in flasks and continuous mode experiments in advanced photobioreactors, it is shown that the limiting nutrient during batch cultivation is sulfate, the depletion of which leads to ROS formation and rapid bleaching of pigments after entry into stationary phase. In continuous mode, however, the limiting nutrient is iron. Optimizing these growth conditions resulted in a so far highest growth rate of 0.16 h -1 (4.3 h doubling time), which is significantly higher than the textbook value of 0.09 h -1 (8 h doubling time). An improved medium, BG-11 for prolonged cultivation (BG-11-PC) is introduced, that allows for controlled, extended cultivation of Synechocystis, under well-defined physiological conditions. The data present here have implications for mass-culturing of cyanobacteria. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Physiological regulation of isocitrate dehydrogenase and the role of 2-oxoglutarate in Prochlorococcus sp. strain PCC 9511.

    Directory of Open Access Journals (Sweden)

    María Agustina Domínguez-Martín

    Full Text Available The enzyme isocitrate dehydrogenase (ICDH; EC 1.1.1.42 catalyzes the oxidative decarboxylation of isocitrate, to produce 2-oxoglutarate. The incompleteness of the tricarboxylic acids cycle in marine cyanobacteria confers a special importance to isocitrate dehydrogenase in the C/N balance, since 2-oxoglutarate can only be metabolized through the glutamine synthetase/glutamate synthase pathway. The physiological regulation of isocitrate dehydrogenase was studied in cultures of Prochlorococcus sp. strain PCC 9511, by measuring enzyme activity and concentration using the NADPH production assay and Western blotting, respectively. The enzyme activity showed little changes under nitrogen or phosphorus starvation, or upon addition of the inhibitors DCMU, DBMIB and MSX. Azaserine, an inhibitor of glutamate synthase, induced clear increases in the isocitrate dehydrogenase activity and icd gene expression after 24 h, and also in the 2-oxoglutarate concentration. Iron starvation had the most significant effect, inducing a complete loss of isocitrate dehydrogenase activity, possibly mediated by a process of oxidative inactivation, while its concentration was unaffected. Our results suggest that isocitrate dehydrogenase responds to changes in the intracellular concentration of 2-oxoglutarate and to the redox status of the cells in Prochlorococcus.

  3. Seawater cultivation of freshwater cyanobacterium Synechocystis sp. PCC 6803 drastically alters amino acid composition and glycogen metabolism

    Directory of Open Access Journals (Sweden)

    Hiroko eIijima

    2015-04-01

    Full Text Available Water use assessment is important for bioproduction using cyanobacteria. For eco-friendly reasons, seawater should preferably be used for cyanobacteria cultivation instead of freshwater. In this study, we demonstrated that the freshwater unicellular cyanobacterium Synechocystis sp. PCC 6803 could be grown in a medium based on seawater. The Synechocystis wild-type strain grew well in an artificial seawater (ASW medium supplemented with nitrogen and phosphorus sources. The addition of HEPES buffer improved cell growth overall, although the growth in ASW medium was inferior to that in the synthetic BG-11 medium. The levels of proteins involved in sugar metabolism changed depending on the culture conditions. The biosynthesis of several amino acids including aspartate, glutamine, glycine, proline, ornithine, and lysine, was highly up-regulated by cultivation in ASW. Two types of natural seawater (NSW were also made available for the cultivation of Synechocystis cells, with supplementation of both nitrogen and phosphorus sources. These results revealed the potential use of seawater for the cultivation of freshwater cyanobacteria, which would help to reduce freshwater consumption during biorefinery using cyanobacteria.

  4. NblA1/A2-Dependent Homeostasis of Amino Acid Pools during Nitrogen Starvation in Synechocystis sp. PCC 6803.

    Science.gov (United States)

    Kiyota, Hiroshi; Hirai, Masami Yokota; Ikeuchi, Masahiko

    2014-06-30

    Nutrient balance is important for photosynthetic growth and biomass production in microalgae. Here, we investigated and compared metabolic responses of amino acid pools to nitrogen and sulfur starvation in a unicellular model cyanobacterium, Synechocystis sp. PCC 6803, and its mutant nblA1/A2. It is known that NblA1/A2-dependent and -independent breakdown of abundant photosynthetic phycobiliproteins and other cellular proteins supply nutrients to the organism. However, the contribution of the NblA1/A2-dependent nutrient supply to amino acid pool homeostasis has not been studied. Our study demonstrates that changes in the pool size of many amino acids during nitrogen starvation can be categorized as NblA1/A2-dependent (Gln, Glu, glutathione, Gly, Ile, Leu, Met, Phe, Pro, Ser, Thr, Tyr and Val) and NblA1/A2-independent (Ala, Asn, Lys, and Trp). We also report unique changes in amino acid pool sizes during sulfur starvation in wild type and the mutant and found a generally marked increase in the Lys pool in cyanobacteria during nutrient starvation. In conclusion, the NblA1/A2-dependent protein turnover contributes to the maintenance of many amino acid pools during nitrogen starvation.

  5. NblA1/A2-Dependent Homeostasis of Amino Acid Pools during Nitrogen Starvation in Synechocystis sp. PCC 6803

    Directory of Open Access Journals (Sweden)

    Hiroshi Kiyota

    2014-06-01

    Full Text Available Nutrient balance is important for photosynthetic growth and biomass production in microalgae. Here, we investigated and compared metabolic responses of amino acid pools to nitrogen and sulfur starvation in a unicellular model cyanobacterium, Synechocystis sp. PCC 6803, and its mutant nblA1/A2. It is known that NblA1/A2-dependent and -independent breakdown of abundant photosynthetic phycobiliproteins and other cellular proteins supply nutrients to the organism. However, the contribution of the NblA1/A2-dependent nutrient supply to amino acid pool homeostasis has not been studied. Our study demonstrates that changes in the pool size of many amino acids during nitrogen starvation can be categorized as NblA1/A2-dependent (Gln, Glu, glutathione, Gly, Ile, Leu, Met, Phe, Pro, Ser, Thr, Tyr and Val and NblA1/A2-independent (Ala, Asn, Lys, and Trp. We also report unique changes in amino acid pool sizes during sulfur starvation in wild type and the mutant and found a generally marked increase in the Lys pool in cyanobacteria during nutrient starvation. In conclusion, the NblA1/A2-dependent protein turnover contributes to the maintenance of many amino acid pools during nitrogen starvation.

  6. Enhanced accumulation of glycogen, lipids and polyhydroxybutyrate under optimal nutrients and light intensities in the cyanobacterium Synechocystis sp. PCC 6803.

    Science.gov (United States)

    Monshupanee, T; Incharoensakdi, A

    2014-04-01

    Glycogen (GL) and lipids (LP) are efficient biofuel substrates, whereas polyhydroxybutyrate (PHB) is a potent biodegradable plastic. This study aimed to increase accumulation of these three compounds in Synechocystis sp. PCC 6803. Under autophototrophic growth, co-accumulation of the three compounds reached maximum level in a mid-stationary phase at 39·2% of dry weight (22·7% GL, 14·1% LP and 2·4% PHB). Nitrogen deprivation increased this to 61·5% (36·8% GL, 11·2% LP and 13·5% PHB), higher than that achieved by phosphorus, sulfur, iron or calcium deprivation. Combining nitrogen deprivation with 0·4% (w/v) glucose addition for heterophototrophic growth and optimizing the light intensity (200 μmol photons m(-2) s(-1) ) synergistically enhanced combined accumulation to 71·1% of biomass (41·3% GL, 16·7% LP and 13·1% PHB), a higher level than previously reported in Synechocystis. However, the maximum coproductivity of GL, LP and PHB (at 0·72 g l(-1) ) was obtained in the 12-day heterophototrophic culture without nitrogen deprivation. Accumulation of GL, LP and PHB was enhanced under both autophototrophic and heterophototrophic conditions by optimizations of nutrient and light supplies. This study provides a means for increasing co-production of potent bioenergy substrates and useful biomaterials in Synechocystis which may also be applicable to other cyanobacteria. © 2013 The Society for Applied Microbiology.

  7. Effects of selected electron transport chain inhibitors on 24-h hydrogen production by Synechocystis sp. PCC 6803.

    Science.gov (United States)

    Burrows, Elizabeth H; Chaplen, Frank W R; Ely, Roger L

    2011-02-01

    One factor limiting biosolar hydrogen (H(2)) production from cyanobacteria is electron availability to the hydrogenase enzyme. In order to optimize 24-h H(2) production this study used Response Surface Methodology and Q2, an optimization algorithm, to investigate the effects of five inhibitors of the photosynthetic and respiratory electron transport chains of Synechocystis sp. PCC 6803. Over 3 days of diurnal light/dark cycling, with the optimized combination of 9.4 mM KCN (3.1 μmol 10(10) cells(-1)) and 1.5 mM malonate (0.5 μmol 10(10) cells(-1)) the H(2) production was 30-fold higher, in EHB-1 media previously optimized for nitrogen (N), sulfur (S), and carbon (C) concentrations (Burrows et al., 2008). In addition, glycogen concentration was measured over 24 h with two light/dark cycling regimes in both standard BG-11 and EHB-1 media. The results suggest that electron flow as well as glycogen accumulation should be optimized in systems engineered for maximal H(2) output. Copyright © 2010 Elsevier Ltd. All rights reserved.

  8. Absorption and emission spectroscopic characterization of blue-light receptor Slr1694 from Synechocystis sp. PCC6803.

    Science.gov (United States)

    Zirak, P; Penzkofer, A; Lehmpfuhl, C; Mathes, T; Hegemann, P

    2007-01-03

    The BLUF protein Slr1694 from the cyanobacterium Synechocystis sp. PCC6803 is characterized by absorption and emission spectroscopy. Slr1694 expressed from E. coli which non-covalently binds FAD, FMN, and riboflavin (called Slr1694(I)), and reconstituted Slr1694 which dominantly contains FAD (called Slr1694(II)) are investigated. The receptor conformation of Slr1694 (dark adapted form Slr1694(r)) is transformed to the putative signalling state (light adapted form Slr1694(s)) with red-shifted absorption and decreased fluorescence efficiency by blue-light excitation. In the dark at 22 degrees C, the signalling state recovers back to the initial receptor state with a time constants of about 14.2s for Slr1694(I) and 17s for Slr1694(II). Quantum yields of signalling state formation of approximately 0.63+/-0.07 for both Slr1694(I) and Slr1694(II) were determined by transient transmission measurements and intensity dependent steady-state transmission measurements. Extended blue-light excitation causes some bound flavin conversion to the hydroquinone form and some photo-degradation, both with low quantum efficiency. The flavin-hydroquinone re-oxidizes slowly back (time constant 5-9 min) to the initial flavoquinone form in the dark. A photo-cycle dynamics scheme is presented.

  9. Absorption and emission spectroscopic characterization of BLUF protein Slr1694 from Synechocystis sp. PCC6803 with roseoflavin cofactor.

    Science.gov (United States)

    Zirak, P; Penzkofer, A; Mathes, T; Hegemann, P

    2009-11-09

    The wild-type BLUF protein Slr1694 from Synechocystis sp. PCC6803 (BLUF=blue-light sensor using FAD) has flavin adenosine dinucleotide (FAD) as natural cofactor. This light sensor causes positive phototaxis of the marine cyanobacterium. In this study the FAD cofactor of the wild-type Slr1694 was replaced by roseoflavin (RoF) and the roseoflavin derivatives RoFMN and RoFAD during heterologous expression in a riboflavin auxotrophic E. coli strain. An absorption and emission spectroscopic characterization of the cofactor-exchanged-Slr1694 (RoSlr) was carried out both under dark conditions and under illuminated conditions. The behaviour of RoF embedded in RoSlr in aqueous solution at pH 8 is compared with the behaviour of RoF in aqueous solution. The fluorescence of RoF and RoSlr is quenched by photo-induced twisted intra-molecular charge transfer at room temperature with stronger effect for RoF. The fluorescence quenching is diminished at liquid nitrogen temperature. Light exposure of RoSlr causes irreversible conversion of the protein embedded roseoflavins to 8-methylamino-flavins, 8-dimethylamino-lumichrome and 8-methylamino-lumichrome.

  10. Amino Acid Transporters and Release of Hydrophobic Amino Acids in the Heterocyst-Forming Cyanobacterium Anabaena sp. Strain PCC 7120

    Directory of Open Access Journals (Sweden)

    Rafael Pernil

    2015-04-01

    Full Text Available Anabaena sp. strain PCC 7120 is a filamentous cyanobacterium that can use inorganic compounds such as nitrate or ammonium as nitrogen sources. In the absence of combined nitrogen, it can fix N2 in differentiated cells called heterocysts. Anabaena also shows substantial activities of amino acid uptake, and three ABC-type transporters for amino acids have been previously characterized. Seven new loci encoding predicted amino acid transporters were identified in the Anabaena genomic sequence and inactivated. Two of them were involved in amino acid uptake. Locus alr2535-alr2541 encodes the elements of a hydrophobic amino acid ABC-type transporter that is mainly involved in the uptake of glycine. ORF all0342 encodes a putative transporter from the dicarboxylate/amino acid:cation symporter (DAACS family whose inactivation resulted in an increased uptake of a broad range of amino acids. An assay to study amino acid release from Anabaena filaments to the external medium was set up. Net release of the alanine analogue α-aminoisobutyric acid (AIB was observed when transport system N-I (a hydrophobic amino acid ABC-type transporter was engaged in the uptake of a specific substrate. The rate of AIB release was directly proportional to the intracellular AIB concentration, suggesting leakage from the cells by diffusion.

  11. CyanoEXpress: A web database for exploration and visualisation of the integrated transcriptome of cyanobacterium Synechocystis sp. PCC6803.

    Science.gov (United States)

    Hernandez-Prieto, Miguel A; Futschik, Matthias E

    2012-01-01

    Synechocystis sp. PCC6803 is one of the best studied cyanobacteria and an important model organism for our understanding of photosynthesis. The early availability of its complete genome sequence initiated numerous transcriptome studies, which have generated a wealth of expression data. Analysis of the accumulated data can be a powerful tool to study transcription in a comprehensive manner and to reveal underlying regulatory mechanisms, as well as to annotate genes whose functions are yet unknown. However, use of divergent microarray platforms, as well as distributed data storage make meta-analyses of Synechocystis expression data highly challenging, especially for researchers with limited bioinformatic expertise and resources. To facilitate utilisation of the accumulated expression data for a wider research community, we have developed CyanoEXpress, a web database for interactive exploration and visualisation of transcriptional response patterns in Synechocystis. CyanoEXpress currently comprises expression data for 3073 genes and 178 environmental and genetic perturbations obtained in 31 independent studies. At present, CyanoEXpress constitutes the most comprehensive collection of expression data available for Synechocystis and can be freely accessed. The database is available for free at http://cyanoexpress.sysbiolab.eu.

  12. Metabolic engineering of the pentose phosphate pathway for enhanced limonene production in the cyanobacterium Synechocysti s sp. PCC 6803.

    Science.gov (United States)

    Lin, Po-Cheng; Saha, Rajib; Zhang, Fuzhong; Pakrasi, Himadri B

    2017-12-13

    Isoprenoids are diverse natural compounds, which have various applications as pharmaceuticals, fragrances, and solvents. The low yield of isoprenoids in plants makes them difficult for cost-effective production, and chemical synthesis of complex isoprenoids is impractical. Microbial production of isoprenoids has been considered as a promising approach to increase the yield. In this study, we engineered the model cyanobacterium Synechocystis sp. PCC 6803 for sustainable production of a commercially valuable isoprenoid, limonene. Limonene synthases from the plants Mentha spicata and Citrus limon were expressed in cyanobacteria for limonene production. Production of limonene was two-fold higher with limonene synthase from M. spicata than that from C. limon. To enhance isoprenoid production, computational strain design was conducted by applying the OptForce strain design algorithm on Synechocystis 6803. Based on the metabolic interventions suggested by this algorithm, genes (ribose 5-phosphate isomerase and ribulose 5-phosphate 3-epimerase) in the pentose phosphate pathway were overexpressed, and a geranyl diphosphate synthase from the plant Abies grandis was expressed to optimize the limonene biosynthetic pathway. The optimized strain produced 6.7 mg/L of limonene, a 2.3-fold improvement in productivity. Thus, this study presents a feasible strategy to engineer cyanobacteria for photosynthetic production of isoprenoids.

  13. Roles of xanthophyll carotenoids in protection against photoinhibition and oxidative stress in the cyanobacterium Synechococcus sp. strain PCC 7002.

    Science.gov (United States)

    Zhu, Yuehui; Graham, Joel E; Ludwig, Marcus; Xiong, Wei; Alvey, Richard M; Shen, Gaozhong; Bryant, Donald A

    2010-12-01

    Synechococcus sp. strain PCC 7002 is a robust, genetically tractable cyanobacterium that produces six different xanthophyll carotenoids (zeaxanthin, cryptoxanthin, myxoxanthophyll (myxol-2'-fucoside), echinenone, 3'-hydroxyechinenone, and synechoxanthin) and tolerates many environmental stresses, including high light intensities. Targeted mutations were introduced to block the branches of the carotenoid biosynthetic pathway leading to specific xanthophylls, and a mutant lacking all xanthophylls was constructed. Some of the mutants showed severe growth defects at high light intensities, and multi-locus mutants had somewhat lower chlorophyll contents and lower photosystem I levels. The results suggested that xanthophylls, particularly zeaxanthin and echinenone, might play regulatory roles in thylakoid biogenesis. Measurements of reactive oxygen (ROS) and nitrogen (RNS) species in the mutants showed that all xanthophylls participate in preventing ROS/RNS accumulation and that a mutant lacking all xanthophylls accumulated very high levels of ROS/RNS. Results from transcription profiling showed that mRNA levels for most genes encoding the enzymes of carotenogenesis are significantly more abundant after exposure to high light. These studies indicated that all xanthophylls contribute to protection against photo-oxidative stress. Copyright © 2010 Elsevier Inc. All rights reserved.

  14. The photosynthetic bacteria Rhodobacter capsulatus and Synechocystis sp. PCC 6803 as new hosts for cyclic plant triterpene biosynthesis.

    Directory of Open Access Journals (Sweden)

    Anita Loeschcke

    Full Text Available Cyclic triterpenes constitute one of the most diverse groups of plant natural products. Besides the intriguing biochemistry of their biosynthetic pathways, plant triterpenes exhibit versatile bioactivities, including antimicrobial effects against plant and human pathogens. While prokaryotes have been extensively used for the heterologous production of other classes of terpenes, the synthesis of cyclic triterpenes, which inherently includes the two-step catalytic formation of the universal linear precursor 2,3-oxidosqualene, is still a major challenge. We thus explored the suitability of the metabolically versatile photosynthetic α-proteobacterium Rhodobacter capsulatus SB1003 and cyanobacterium Synechocystis sp. PCC 6803 as alternative hosts for biosynthesis of cyclic plant triterpenes. Therefore, 2,3-oxidosqualene production was implemented and subsequently combined with different cyclization reactions catalyzed by the representative oxidosqualene cyclases CAS1 (cycloartenol synthase, LUP1 (lupeol synthase, THAS1 (thalianol synthase and MRN1 (marneral synthase derived from model plant Arabidopsis thaliana. While successful accumulation of 2,3-oxidosqualene could be detected by LC-MS analysis in both hosts, cyclase expression resulted in differential production profiles. CAS1 catalyzed conversion to only cycloartenol, but expression of LUP1 yielded lupeol and a triterpenoid matching an oxidation product of lupeol, in both hosts. In contrast, THAS1 expression did not lead to cyclic product formation in either host, whereas MRN1-dependent production of marnerol and hydroxymarnerol was observed in Synechocystis but not in R. capsulatus. Our findings thus indicate that 2,3-oxidosqualene cyclization in heterologous phototrophic bacteria is basically feasible but efficient conversion depends on both the respective cyclase enzyme and individual host properties. Therefore, photosynthetic α-proteo- and cyanobacteria are promising alternative candidates

  15. Ethylene Regulates the Physiology of the Cyanobacterium Synechocystis sp. PCC 6803 via an Ethylene Receptor1[OPEN

    Science.gov (United States)

    2016-01-01

    Ethylene is a plant hormone that plays a crucial role in the growth and development of plants. The ethylene receptors in plants are well studied, and it is generally assumed that they are found only in plants. In a search of sequenced genomes, we found that many bacterial species contain putative ethylene receptors. Plants acquired many proteins from cyanobacteria as a result of the endosymbiotic event that led to chloroplasts. We provide data that the cyanobacterium Synechocystis (Synechocystis sp. PCC 6803) has a functional receptor for ethylene, Synechocystis Ethylene Response1 (SynEtr1). We first show that SynEtr1 directly binds ethylene. Second, we demonstrate that application of ethylene to Synechocystis cells or disruption of the SynEtr1 gene affects several processes, including phototaxis, type IV pilus biosynthesis, photosystem II levels, biofilm formation, and spontaneous cell sedimentation. Our data suggest a model where SynEtr1 inhibits downstream signaling and ethylene inhibits SynEtr1. This is similar to the inverse-agonist model of ethylene receptor signaling proposed for plants and suggests a conservation of structure and function that possibly originated over 1 billion years ago. Prior research showed that SynEtr1 also contains a light-responsive phytochrome-like domain. Thus, SynEtr1 is a bifunctional receptor that mediates responses to both light and ethylene. To our knowledge, this is the first demonstration of a functional ethylene receptor in a nonplant species and suggests that that the perception of ethylene is more widespread than previously thought. PMID:27246094

  16. Specific role of the cyanobacterial PipX factor in the heterocysts of Anabaena sp. strain PCC 7120.

    Science.gov (United States)

    Valladares, Ana; Rodríguez, Virginia; Camargo, Sergio; Martínez-Noël, Giselle M A; Herrero, Antonia; Luque, Ignacio

    2011-03-01

    The PipX factor is a regulatory protein that seems to occur only in cyanobacteria. In the filamentous, heterocyst-forming Anabaena sp. strain PCC 7120, open reading frame (ORF) asr0485, identified as the pipX gene, is expressed mainly under conditions of combined-nitrogen deprivation dependent on the global N regulator NtcA and the heterocyst-specific regulator HetR. Primer extension and 5' rapid amplification of cDNA ends (RACE) analyses detected three transcription start points corresponding to a canonical NtcA-activated promoter (to which direct binding of NtcA was observed), an NtcA- and HetR-dependent promoter, and a consensus-type promoter, the last with putative -35 and -10 determinants. Activation of pipX took place in cells differentiating into heterocysts at intermediate to late stages of the process. Accordingly, disruption of pipX led to impaired diazotrophic growth, reduced nitrogenase activity, and impaired activation of the nitrogenase structural genes. The nitrogenase activity of the mutant was low under oxic conditions, likely resulting from inefficient protection against oxygen. In line with this, the activation of the coxB2A2C2 and coxB3A3C3 operons, encoding heterocyst-specific terminal respiratory oxidases responsible for internal oxygen removal, was deficient in the pipX mutant. Therefore, the Anabaena PipX factor shows a spatiotemporal specificity contributing to normal heterocyst function, including full activation of the nitrogenase structural genes and genes of the nitrogenase-protective features of the heterocyst.

  17. Identification of OmpR-family response regulators interacting with thioredoxin in the Cyanobacterium Synechocystis sp. PCC 6803.

    Directory of Open Access Journals (Sweden)

    Taro Kadowaki

    Full Text Available The redox state of the photosynthetic electron transport chain is known to act as a signal to regulate the transcription of key genes involved in the acclimation responses to environmental changes. We hypothesized that the protein thioredoxin (Trx acts as a mediator connecting the redox state of the photosynthetic electron transport chain and transcriptional regulation, and established a screening system to identify transcription factors (TFs that interact with Trx. His-tagged TFs and S-tagged mutated form of Trx, TrxMC35S, whose active site cysteine 35 was substituted with serine to trap the target interacting protein, were co-expressed in E. coli cells and Trx-TF complexes were detected by immuno-blotting analysis. We examined the interaction between Trx and ten OmpR family TFs encoded in the chromosome of the cyanobacterium Synechocystis sp. PCC 6803 (S.6803. Although there is a highly conserved cysteine residue in the receiver domain of all OmpR family TFs, only three, RpaA (Slr0115, RpaB (Slr0947 and ManR (Slr1837, were identified as putative Trx targets [corrected].The recombinant forms of wild-type TrxM, RpaA, RpaB and ManR proteins from S.6803 were purified following over-expression in E. coli and their interaction was further assessed by monitoring changes in the number of cysteine residues with free thiol groups. An increase in the number of free thiols was observed after incubation of the oxidized TFs with Trx, indicating the reduction of cysteine residues as a consequence of interaction with Trx. Our results suggest, for the first time, the possible regulation of OmpR family TFs through the supply of reducing equivalents from Trx, as well as through the phospho-transfer from its cognate sensor histidine kinase.

  18. CopM is a novel copper-binding protein involved in copper resistance in Synechocystis sp. PCC 6803

    Science.gov (United States)

    Giner-Lamia, Joaquín; López-Maury, Luis; Florencio, Francisco J

    2015-01-01

    Copper resistance system in the cyanobacterium Synechocystis sp. PCC 6803 comprises two operons, copMRS and copBAC, which are expressed in response to copper in the media. copBAC codes for a heavy-metal efflux–resistance nodulation and division (HME-RND) system, while copMRS codes for a protein of unknown function, CopM, and a two-component system CopRS, which controls the expression of these two operons. Here, we report that CopM is a periplasmic protein able to bind Cu(I) with high affinity (KD ∼3 × 10−16). Mutants lacking copM showed a sensitive copper phenotype similar to mutants affected in copB, but lower than mutants of the two-component system CopRS, suggesting that CopBAC and CopM constitute two independent resistance mechanisms. Moreover, constitutive expression of copM is able to partially suppress the copper sensitivity of the copR mutant strain, pointing out that CopM per se is able to confer copper resistance. Furthermore, constitutive expression of copM was able to reduce total cellular copper content of the copR mutant to the levels determined in the wild-type (WT) strain. Finally, CopM was localized not only in the periplasm but also in the extracellular space, suggesting that CopM can also prevent copper accumulation probably by direct copper binding outside the cell. PMID:25545960

  19. Photoautotrophic Polyhydroxybutyrate Granule Formation Is Regulated by Cyanobacterial Phasin PhaP in Synechocystis sp. Strain PCC 6803

    Science.gov (United States)

    Hauf, Waldemar; Watzer, Björn; Roos, Nora; Klotz, Alexander

    2015-01-01

    Cyanobacteria are photoautotrophic microorganisms which fix atmospheric carbon dioxide via the Calvin-Benson cycle to produce carbon backbones for primary metabolism. Fixed carbon can also be stored as intracellular glycogen, and in some cyanobacterial species like Synechocystis sp. strain PCC 6803, polyhydroxybutyrate (PHB) accumulates when major nutrients like phosphorus or nitrogen are absent. So far only three enzymes which participate in PHB metabolism have been identified in this organism, namely, PhaA, PhaB, and the heterodimeric PHB synthase PhaEC. In this work, we describe the cyanobacterial PHA surface-coating protein (phasin), which we term PhaP, encoded by ssl2501. Translational fusion of Ssl2501 with enhanced green fluorescent protein (eGFP) showed a clear colocalization to PHB granules. A deletion of ssl2501 reduced the number of PHB granules per cell, whereas the mean PHB granule size increased as expected for a typical phasin. Although deletion of ssl2501 had almost no effect on the amount of PHB, the biosynthetic activity of PHB synthase was negatively affected. Secondary-structure prediction and circular dichroism (CD) spectroscopy of PhaP revealed that the protein consists of two α-helices, both of them associating with PHB granules. Purified PhaP forms oligomeric structures in solution, and both α-helices of PhaP contribute to oligomerization. Together, these results support the idea that Ssl2501 encodes a cyanobacterial phasin, PhaP, which regulates the surface-to-volume ratio of PHB granules. PMID:25911471

  20. Lipid and carotenoid cooperation-driven adaptation to light and temperature stress in Synechocystis sp. PCC6803.

    Science.gov (United States)

    Zakar, Tomas; Herman, Eva; Vajravel, Sindhujaa; Kovacs, Laszlo; Knoppová, Jana; Komenda, Josef; Domonkos, Ildiko; Kis, Mihaly; Gombos, Zoltan; Laczko-Dobos, Hajnalka

    2017-05-01

    Polyunsaturated lipids are important components of photosynthetic membranes. Xanthophylls are the main photoprotective agents, can assist in protection against light stress, and are crucial in the recovery from photoinhibition. We generated the xanthophyll- and polyunsaturated lipid-deficient ROAD mutant of Synechocystis sp. PCC6803 (Synechocystis) in order to study the little-known cooperative effects of lipids and carotenoids (Cars). Electron microscopic investigations confirmed that in the absence of xanthophylls the S-layer of the cellular envelope is missing. In wild-type (WT) cells, as well as the xanthophyll-less (RO), polyunsaturated lipid-less (AD), and the newly constructed ROAD mutants the lipid and Car compositions were determined by MS and HPLC, respectively. We found that, relative to the WT, the lipid composition of the mutants was remodeled and the Car content changed accordingly. In the mutants the ratio of non-bilayer-forming (NBL) to bilayer-forming (BL) lipids was found considerably lower. Xanthophyll to β-carotene ratio increased in the AD mutant. In vitro and in vivo methods demonstrated that saturated, monounsaturated lipids and xanthophylls may stabilize the trimerization of Photosystem I (PSI). Fluorescence induction and oxygen-evolving activity measurements revealed increased light sensitivity of RO cells compared to those of the WT. ROAD showed a robust increase in light susceptibility and reduced recovery capability, especially at moderate low (ML) and moderate high (MH) temperatures, indicating a cooperative effect of xanthophylls and polyunsaturated lipids. We suggest that both lipid unsaturation and xanthophylls are required for providing the proper structure and functioning of the membrane environment that protects against light and temperature stress. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Crescimento mixotrófico de Nostoc sp. Glucose, sacarose e melaço de cana-de-açúcar foram testados como substratos para produção de biomassa e ficobiliproteinas - DOI: 10.4025/actascibiolsci.v29i1.121 Mixotrophic growth of Nostoc sp. on glucose, sucrose and sugarcane molasses for phycobiliprotein production

    Directory of Open Access Journals (Sweden)

    Maria Helena Pimenta Pinotti

    2007-11-01

    Full Text Available Todos os substratos aumentaram a biomassa e ficobiliproteinas em relação ao controle, meio mineral BG11. Melaço de cana-de-açúcar foi o melhor substrato tanto para a produção de biomassa como de ficobiliproteinas. A maior produção de biomassa ocorreu usando melaço de cana-de-açúcar 1,0 g L-1 sendo 5,7 vezes maior que o controle. Com glucose foi em 2,5 g L-1 e sacarose 1,5 g L-1, sendo 2,5 e 4,8 vezes maior que o controle, respectivamente. A maior produção de ficobiliproteinas ocorreu usando melaço de cana-de-açúcar 1,0 g L-1 sendo 12,5 vezes maior que o controle. Com glucose foi em 1,0 g L-1 e sacarose 0,5 g L-1, 3,0 e 4,5 vezes maior que o controle, respectivamente. Nostoc sp. testado pode crescer mixotroficamente, usando glucose, sacarose e melaço de cana-deaçúcar como substratos orgânicos, uma maior produção de biomassa e ficobiliproteinas podendo ser alcançada nessas condições quando comparadas com o crescimento autotrófico.Glucose, sacarose, and sugarcane molasses were tested as substrates for production of biomass and phycobiliproteins by Nostoc sp., varying their concentrations in relation to a mineral medium, BG11. All substrates increased the biomass and phycobiliproteins when compared with the control. Sugarcane molasses showed to be the best substrate for production of both biomass and phycobiliproteins. Greater biomass production occurred in sugarcane molasses 1.0 g L-1 and it was 5.7 times greater than the control. With glucose, it was in 2.5 g L-1 and sucrose, in 1.5 g L-1, reaching 2.5 and 4.8 times greater than the control, respectively. For phycobiliproteins, the major production was in sugarcane molasses 1.0 g L-1, 12.5 times greater than the control. With glucose, it was in 1.0 g L-1 and sucrose, in 0,5 g L-1, reaching 3.0 and 4.5 times greater than the control, respectively. The Nostoc sp. assayed can grow mixotrophically, using glucose, sucrose, and sugarcane molasses as organic substrates, and a

  2. Mixotrophic growth of Nostoc sp. on glucose, sucrose and sugarcane molasses for phycobiliprotein production = Crescimento mixotrófico de Nostoc sp. Glucose, sacarose e melaço de cana-de-açúcar foram testados como substratos para produção de biomassa e ficobiliproteinas

    Directory of Open Access Journals (Sweden)

    Maria Helena Pimenta Pinotti

    2007-01-01

    Full Text Available Glucose, sacarose, and sugarcane molasses were tested as substrates for production of biomass and phycobiliproteins by Nostoc sp., varying their concentrations in relation to a mineral medium, BG11. All substrates increased the biomass and phycobiliproteins when compared with the control. Sugarcane molasses showed to be thebest substrate for production of both biomass and phycobiliproteins. Greater biomass production occurred in sugarcane molasses 1.0 g L-1 and it was 5.7 times greater than the control. With glucose, it was in 2.5 g L-1 and sucrose, in 1.5 g L-1, reaching 2.5 and 4.8 timesgreater than the control, respectively. For phycobiliproteins, the major production was in sugarcane molasses 1.0 g L-1, 12.5 times greater than the control. With glucose, it was in 1.0 g L-1 and sucrose, in 0,5 g L-1, reaching 3.0 and 4.5 times greater than the control, respectively. The Nostoc sp. assayed can grow mixotrophically, using glucose, sucrose, and sugarcane molasses as organic substrates, and a greater production of biomass andphycobiliproteins can be reached when compared with the autotrophic growth.Todos os substratos aumentaram a biomassa e ficobiliproteinas emrelação ao controle, meio mineral BG11. Melaço de cana-de-açúcar foi o melhor substrato tanto para a produção de biomassa como de ficobiliproteinas. A maior produção de biomassa ocorreu usando melaço de cana-de-açúcar 1,0 g L-1 sendo 5,7 vezes maior que o controle. Com glucose foi em 2,5 g L-1 e sacarose 1,5 g L-1, sendo 2,5 e 4,8 vezes maior que o controle, respectivamente. A maior produção de ficobiliproteinas ocorreu usando melaço de cana-de-açúcar 1,0 g L-1 sendo 12,5 vezes maior que o controle. Com glucose foi em 1,0g L-1 e sacarose 0,5 g L-1, 3,0 e 4,5 vezes maior que o controle, respectivamente. Nostoc sp. testado pode crescer mixotroficamente, usando glucose, sacarose e melaço de cana-deaçúcar como substratos orgânicos, uma maior produção de biomassa e

  3. Mutation of the murC and murB Genes Impairs Heterocyst Differentiation in Anabaena sp. Strain PCC 7120.

    Science.gov (United States)

    Videau, Patrick; Rivers, Orion S; Ushijima, Blake; Oshiro, Reid T; Kim, Min Joo; Philmus, Benjamin; Cozy, Loralyn M

    2016-04-01

    To stabilize cellular integrity in the face of environmental perturbations, most bacteria, including cyanobacteria, synthesize and maintain a strong, flexible, three-dimensional peptidoglycan lattice. Anabaena sp. strain PCC 7120 is a filamentous cyanobacterium capable of differentiating morphologically distinct nitrogen-fixing heterocyst cells in a periodic pattern. While heterocyst development has been shown to require proper peptidoglycan remodeling, the role of peptidoglycan synthesis has remained unclear. Here we report the identification of two peptidoglycan synthesis genes, murC (alr5065) and murB (alr5066), as required for heterocyst development. The murC and murB genes are predicted to encode a UDP-N-acetylmuramate:L-alanine ligase and a UDP-N-acetylenolpyruvoylglucosamine reductase, respectively, and we confirm enzymatic function through complementation of Escherichia coli strains deficient for these enzymes. Cells depleted of either murC or murB expression failed to differentiate heterocysts under normally inducing conditions and displayed decreased filament integrity. To identify the stage(s) of development affected by murC or murB depletion, the spatial distribution of expression of the patterning marker gene, patS, was examined. Whereas murB depletion did not affect the pattern of patS expression, murC depletion led to aberrant expression of patS in all cells of the filament. Finally, expression of gfp controlled by the region of DNA immediately upstream of murC was enriched in differentiating cells and was repressed by the transcription factor NtcA. Collectively, the data in this work provide evidence for a direct link between peptidoglycan synthesis and the maintenance of a biological pattern in a multicellular organism. Multicellular organisms that differentiate specialized cells must regulate morphological changes such that both cellular integrity and the dissemination of developmental signals are preserved. Here we show that the multicellular

  4. Co-ordination of NDH and Cup proteins in CO2 uptake in cyanobacterium Synechocystis sp. PCC 6803.

    Science.gov (United States)

    Han, Xunling; Sun, Nan; Xu, Min; Mi, Hualing

    2017-06-01

    High and low affinity CO2-uptake systems containing CupA (NDH-1MS) and CupB (NDH-1MS'), respectively, have been identified in Synechocystis sp. PCC 6803, but it is yet unknown how the complexes function in CO2 uptake. In this work, we found that deletion of cupB significantly lowered the growth of cells, and deletion of both cupA and cupB seriously suppressed the growth below pH 7.0 even under 3% CO2. The rate of photosynthetic oxygen evolution was decreased slightly by deletion of cupA but significantly by deletion of cupB and more severely by deletion of both cupA and cupB, especially in response to changed pH conditions under 3% CO2. Furthermore, we found that assembly of CupB into NDH-1MS' was dependent on NdhD4 and NdhF4. NDH-1MS' was not affected in the NDH-1MS-degradation mutant and NDH-1MS was not affected in the NDH-1MS'-degradation mutants, indicating the existence of independent CO2-uptake systems under high CO2 conditions. The light-induced proton gradient across thylakoid membranes was significantly inhibited in ndhD-deletion mutants, suggesting that NdhDs functions in proton pumping. The carbonic anhydrase activity was suppressed partly in the cupA- or cupB-deletion mutant but severely in the mutant with both cupA and cupB deletion, indicating that CupA and CupB function in conversion of CO2 to HCO3-. In turn, deletion of cup genes lowered the transthylakoid membrane proton gradient and deletion of ndhDs decreased the CO2 hydration. Our results suggest that NDH-1M provides an alkaline region to activate Cup proteins involved in CO2 uptake. © The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  5. Spectral and Temporal Properties of the Alpha and Beta Subunits and (alpha Beta) Monomer Isolated from Nostoc SP. Using Picosecond Laser Spectroscopy.

    Science.gov (United States)

    Dagen, Aaron J.

    1985-12-01

    The fluorescence decay profiles, relative quantum yield and transmission of the (alpha), (beta) and ((alpha)(beta)) complexes from phycoerythrin isolated from the photosynthetic antenna system of Nostoc sp. and measured by single picosecond laser spectroscopic techniques is studied. The fluorescence decay profiles of all three complexes are found to be intensity independent for the intensity range investigated ((TURN)4 x 10('13) to (TURN)4 x 10('15) photons-cm('-2) per pulse). The apparent decrease in the relative quantum yield of all three complexes as intensity increases is offset by a corresponding increase in the relative transmission. This evidence, along with the intensity independent fluorescence kinetics, suggests that exciton annihilation is absent in these complexes. The decay profiles are fit to models assuming energy transfer amongst fluorescing chromophores. The intraprotein transfer rate is found to be 100 ps in the (alpha) subunit, 666 ps in the (beta) subunit. Constraining these rates to be identical in the monomer results in explaining the monomer kinetics by an increase in the nonradiative rate of the f(,(beta)) chromophore, an apparent result of aggregation effects.

  6. Protein (Cyanobacteria): 130081 [PGDBj - Ortholog DB

    Lifescience Database Archive (English)

    Full Text Available n all1672 Nostoc sp. PCC 7120 MFKILFDSDLILDAVMNRTELAEDVRTLLENLHPSIRLYLTDVGLQKVSTYTYCLKNSQIPEIIVDWLQEQIQICPIDQGLLQKARYSPLRDFESAVELACINHYQLNAIVTNKPEDFIVTAHPLCVWSFADLWLRVNLESQLQATIHS ...

  7. Protein (Cyanobacteria): 464885 [PGDBj - Ortholog DB

    Lifescience Database Archive (English)

    Full Text Available tein Nos7524_5075 Nostoc sp. PCC 7524 MSLSYDISSILNLLRSLPSTELRTVKEEIDSILKERGTTIRIPDPFKIVPAQVVLKDSNLEESTSEVKLEEEYQQINEDISEPSGVLNLSSIKDATDNKAEKKEAIQEIPRPLGIWKGKVEISEDFYETTNDILSEFGIEE ...

  8. ORF Sequence: NC_003272 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available rotein [Nostoc sp. PCC 7120] MALMGATGSGKSTLLENLIGIKQPQSGKIWINDISLEPQTLPQVRRYIGFGFQDANDQLFMPTILEDITFGPLNYGVPAAIARDQARQLLADFGLEAYANRSAH...ELSGGQRRLAALAAILALEPAILILDEPTTGLDPAWRRHLARVLFNLPVQVMLIASHELHWLGKVTQRALVLSNGRIQLDNEIQPLLQNGEILEQLGLPIDW

  9. Sll0528, a Site-2-Protease, Is Critically Involved in Cold, Salt and Hyperosmotic Stress Acclimation of Cyanobacterium Synechocystis sp. PCC 6803

    Directory of Open Access Journals (Sweden)

    Haijin Lei

    2014-12-01

    Full Text Available Site-2-proteases (S2Ps mediated proteolysis of transmembrane transcriptional regulators is a conserved mechanism to regulate transmembrane signaling. The universal presence of S2P homologs in different cyanobacterial genomes suggest conserved and fundamental functions, though limited data has been available. Here we provide the first evidence that Sll0528, a site-2-protease in Synechocystis sp. PCC 6803 is crucial for salt, cold and hyperosmotic stress acclimation. Remarkable induction of sll0528 gene expression was observed under salt, cold and hyperosmotic stress, much higher than induction of the other three S2Ps. Knock-out of sll0528 gene in wild type Synechocystis sp. PCC 6803 increased their sensitivity to salt, cold and hyperosmotic stress, as revealed by retarded growth, reduced pigments and disrupted photosystems. The sll0528 gene was induced to a much smaller extent by high light and mixotrophic growth with glucose. Similar growth responses of the sll0528 knockout mutant and wild type under high light and mixotrophic growth indicated that sll0528 was dispensable for these conditions. Recombinant Sll0528 protein could cleave beta-casein into smaller fragments. These results together suggest that the Sll0528 metalloprotease plays a role in the stress response and lays the foundation for further investigation of its mechanism, as well as providing hints for the functional analysis of other S2Ps in cyanobacteria.

  10. Integrated in silico Analyses of Regulatory and Metabolic Networks of Synechococcus sp. PCC 7002 Reveal Relationships between Gene Centrality and Essentiality

    Directory of Open Access Journals (Sweden)

    Hyun-Seob Song

    2015-03-01

    Full Text Available Cyanobacteria dynamically relay environmental inputs to intracellular adaptations through a coordinated adjustment of photosynthetic efficiency and carbon processing rates. The output of such adaptations is reflected through changes in transcriptional patterns and metabolic flux distributions that ultimately define growth strategy. To address interrelationships between metabolism and regulation, we performed integrative analyses of metabolic and gene co-expression networks in a model cyanobacterium, Synechococcus sp. PCC 7002. Centrality analyses using the gene co-expression network identified a set of key genes, which were defined here as “topologically important.” Parallel in silico gene knock-out simulations, using the genome-scale metabolic network, classified what we termed as “functionally important” genes, deletion of which affected growth or metabolism. A strong positive correlation was observed between topologically and functionally important genes. Functionally important genes exhibited variable levels of topological centrality; however, the majority of topologically central genes were found to be functionally essential for growth. Subsequent functional enrichment analysis revealed that both functionally and topologically important genes in Synechococcus sp. PCC 7002 are predominantly associated with translation and energy metabolism, two cellular processes critical for growth. This research demonstrates how synergistic network-level analyses can be used for reconciliation of metabolic and gene expression data to uncover fundamental biological principles.

  11. Identification of a New Target slr0946 of the Response Regulator Sll0649 Involving Cadmium Tolerance in Synechocystis sp. PCC 6803

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    Tao Sun

    2017-08-01

    Full Text Available Survival of photosynthetic cyanobacteria is challenged by environmental contaminations like heavy metals. Among them, deciphering the regulatory mechanisms for cadmium (Cd in cyanobacteria would facilitate the construction of Cd-resistant strains. In this study, the DNA-Affinity-Purified-chromatin immunoprecipitation assay was employed to identify the direct targets of Sll0649, which was a Cd2+-related response regulator identified in our previous work in model cyanobacteria Synechocystis sp. PCC 6803. As a result, the promoter region of slr0946 encoding the arsenate reductase was enriched fourfolds by quantitative real time PCR analysis. Further, deletion of slr0946 led to a sensitive phenotype to Cd2+ stress compared with the wild type (WT and the sensitive phenotype of Δslr0946 could be rescued by complementation assay via introducing slr0946 back into Δslr0946. Finally, individually overexpression of slr0946 as well as two Cd2+-related genes identified priviously (i.e., sll1598 and slr0798 in WT could significantly improve the tolerance of Synechocystis sp. PCC 6803 to Cd2+. This study provided a better understanding of the tolerance mechanism to Cd2+ in cyanobacteria and also feasible strategies for tolerance modifications to heavy metals in the future.

  12. The Multiple Functions of Common Microbial Carbon Polymers, Glycogen and PHB, during Stress Responses in the Non-Diazotrophic Cyanobacterium Synechocystis sp. PCC 6803.

    Science.gov (United States)

    Damrow, Ramon; Maldener, Iris; Zilliges, Yvonne

    2016-01-01

    Classical microbial carbon polymers such as glycogen and polyhydroxybutyrate (PHB) have a crucial impact as both a sink and a reserve under macronutrient stress conditions. Most microbial species exclusively synthesize and degrade either glycogen or PHB. A few bacteria such as the phototrophic model organism Synechocystis sp. PCC 6803 surprisingly produce both physico-chemically different polymers under conditions of high C to N ratios. For the first time, the function and interrelation of both carbon polymers in non-diazotrophic cyanobacteria are analyzed in a comparative physiological study of single- and double-knockout mutants (ΔglgC; ΔphaC; ΔglgC/ΔphaC), respectively. Most of the observed phenotypes are explicitly related to the knockout of glycogen synthesis, highlighting the metabolic, energetic, and structural impact of this process whenever cells switch from an active, photosynthetic 'protein status' to a dormant 'glycogen status'. The carbon flux regulation into glycogen granules is apparently crucial for both phycobilisome degradation and thylakoid layer disassembly in the presence of light. In contrast, PHB synthesis is definitely not involved in this primary acclimation response. Moreover, the very weak interrelations between the two carbon-polymer syntheses indicate that the regulation and role of PHB synthesis in Synechocystis sp. PCC 6803 is different from glycogen synthesis.

  13. The interplay between siderophore secretion and coupled iron and copper transport in the heterocyst-forming cyanobacterium Anabaena sp. PCC 7120.

    Science.gov (United States)

    Nicolaisen, Kerstin; Hahn, Alexander; Valdebenito, Marianne; Moslavac, Suncana; Samborski, Anastazia; Maldener, Iris; Wilken, Corinna; Valladares, Ana; Flores, Enrique; Hantke, Klaus; Schleiff, Enrico

    2010-11-01

    Iron uptake is essential for Gram-negative bacteria including cyanobacteria. In cyanobacteria, however, the iron demand is higher than in proteobacteria due to the function of iron as a cofactor in photosynthesis and nitrogen fixation, but our understanding of iron uptake by cyanobacteria stands behind the knowledge in proteobacteria. Here, two genes involved in this process in the heterocyst-forming cyanobacterium Anabaena sp. PCC 7120 were identified. ORF all4025 encodes SchE, a putative cytoplasmic membrane-localized transporter involved in TolC-dependent siderophore secretion. Inactivation of schE resulted in an enhanced sensitivity to high metal concentrations and decreased secretion of hydroxamate-type siderophores. ORF all4026 encodes a predicted outer membrane-localized TonB-dependent iron transporter, IacT. Inactivation of iacT resulted in decreased sensitivity to elevated iron and copper levels. Expression of iacT from the artificial trc promoter (P(trc)) resulted in sensitization against tested metals. Further analysis showed that iron and copper effects are synergistic because a decreased supply of iron induced a significant decrease of copper levels in the iacT insertion mutant but an increase of those levels in the strain carrying P(trc)-iacT. Our results unravel a link between iron and copper homeostasis in Anabaena sp. PCC 7120. Copyright © 2010 Elsevier B.V. All rights reserved.

  14. An alternative methionine aminopeptidase, MAP-A, is required for nitrogen starvation and high-light acclimation in the cyanobacterium Synechocystis sp. PCC 6803.

    Science.gov (United States)

    Drath, Miriam; Baier, Kerstin; Forchhammer, Karl

    2009-05-01

    Methionine aminopeptidases (MetAPs or MAPs, encoded by map genes) are ubiquitous and pivotal enzymes for protein maturation in all living organisms. Whereas most bacteria harbour only one map gene, many cyanobacterial genomes contain two map paralogues, the genome of Synechocystis sp. PCC 6803 even three. The physiological function of multiple map paralogues remains elusive so far. This communication reports for the first time differential MetAP function in a cyanobacterium. In Synechocystis sp. PCC 6803, the universally conserved mapC gene (sll0555) is predominantly expressed in exponentially growing cells and appears to be a housekeeping gene. By contrast, expression of mapA (slr0918) and mapB (slr0786) genes increases during stress conditions. The mapB paralogue is only transiently expressed, whereas the widely distributed mapA gene appears to be the major MetAP during stress conditions. A mapA-deficient Synechocystis mutant shows a subtle impairment of photosystem II properties even under non-stressed conditions. In particular, the binding site for the quinone Q(B) is affected, indicating specific N-terminal methionine processing requirements of photosystem II components. MAP-A-specific processing becomes essential under certain stress conditions, since the mapA-deficient mutant is severely impaired in surviving conditions of prolonged nitrogen starvation and high light exposure.

  15. Overexpression of FurA in Anabaena sp. PCC 7120 reveals new targets for this regulator involved in photosynthesis, iron uptake and cellular morphology.

    Science.gov (United States)

    González, Andrés; Bes, M Teresa; Barja, François; Peleato, M Luisa; Fillat, María F

    2010-11-01

    Previous genomic analyses of the filamentous nitrogen-fixing cyanobacterium Anabaena sp. PCC 7120 have identified three ferric uptake regulator (Fur) homologs with low sequence identities and probably different functions in the cell. FurA is a constitutive protein that shares the highest homology with Fur from heterotrophic bacteria and appears to be essential for in vitro growth. In this study, we have analysed the effects of FurA overexpression on the Anabaena sp. phenotype and investigated which of the observed alterations were directly operated by FurA. Overexpression of the regulator led to changes in cellular morphology, resulting in shorter filaments with rounded cells of different sizes. The furA-overexpressing strain showed a slower photoautotrophic growth and a marked decrease in the oxygen evolution rate. Overexpression of the regulator also decreased both catalase and superoxide dismutase activities, but did not lead to an increase in the levels of intracellular reactive oxygen species. By combining phenotypic studies, reverse transcription-PCR analyses and electrophoretic mobility shift assays, we identified three novel direct targets of FurA, including genes encoding a siderophore outer membrane transporter (schT), bacterial actins (mreBCD) and the PSII reaction center protein D1 (psbA). The affinity of FurA for these novel targets was markedly affected by the absence of divalent metal ions, confirming previous evidence of a critical role for the metal co-repressor in the function of the regulator in vivo. The results unravel new cellular processes modulated by FurA, supporting its role as a global transcriptional regulator in Anabaena sp. PCC 7120.

  16. Regulation of intracellular free calcium concentration during heterocyst differentiation by HetR and NtcA in Anabaena sp. PCC 7120.

    Science.gov (United States)

    Shi, Yunming; Zhao, Weixing; Zhang, Wei; Ye, Zi; Zhao, Jindong

    2006-07-25

    Calcium ions are important to some prokaryotic cellular processes, such as heterocyst differentiation of cyanobacteria. Intracellular free Ca(2+)concentration, [Ca(2+)](i), increases several fold in heterocysts and is regulated by CcbP, a Ca(2+)-binding protein found in heterocyst-forming cyanobacteria. We demonstrate here that CcbP is degraded by HetR, a serine-type protease that controls heterocyst differentiation. The degradation depends on Ca(2+) and appears to be specific because HetR did not digest other tested proteins. CcbP was found to bind two Ca(2+) per molecule with K(D) values of 200 nM and 12.8 microM. Degradation of CcbP releases bound Ca(2+) that contributes significantly to the increase of [Ca(2+)](i) during the process of heterocyst differentiation in Anabaena sp. strain PCC 7120. We suggest that degradation of CcbP is a mechanism of positive autoregulation of HetR. The down-regulation of ccbP in differentiating cells and mature heterocysts, which also is critical to the regulation of [Ca(2+)](i), depends on NtcA. Coexpression of ntcA and a ccbP promoter-controlled gfp in Escherichia coli diminished production of GFP, and the decrease is enhanced by alpha-ketoglutarate. It was also found that NtcA could bind a fragment of the ccbP promoter containing an NtcA-binding sequence in a alpha-ketoglutarate-dependent fashion. Therefore, [Ca(2+)](i) is regulated by a collaboration of HetR and NtcA in heterocyst differentiation in Anabaena sp. strain PCC 7120.

  17. Optimization of pH and nitrogen for enhanced hydrogen production by Synechocystis sp. PCC 6803 via statistical and machine learning methods.

    Science.gov (United States)

    Burrows, Elizabeth H; Wong, Weng-Keen; Fern, Xiaoli; Chaplen, Frank W R; Ely, Roger L

    2009-01-01

    The nitrogen (N) concentration and pH of culture media were optimized for increased fermentative hydrogen (H(2)) production from the cyanobacterium, Synechocystis sp. PCC 6803. The optimization was conducted using two procedures, response surface methodology (RSM), which is commonly used, and a memory-based machine learning algorithm, Q2, which has not been used previously in biotechnology applications. Both RSM and Q2 were successful in predicting optimum conditions that yielded higher H(2) than the media reported by Burrows et al., Int J Hydrogen Energy. 2008;33:6092-6099 optimized for N, S, and C (called EHB-1 media hereafter), which itself yielded almost 150 times more H(2) than Synechocystis sp. PCC 6803 grown on sulfur-free BG-11 media. RSM predicted an optimum N concentration of 0.63 mM and pH of 7.77, which yielded 1.70 times more H(2) than EHB-1 media when normalized to chlorophyll concentration (0.68 +/- 0.43 micromol H(2) mg Chl(-1) h(-1)) and 1.35 times more when normalized to optical density (1.62 +/- 0.09 nmol H(2) OD(730) (-1) h(-1)). Q2 predicted an optimum of 0.36 mM N and pH of 7.88, which yielded 1.94 and 1.27 times more H(2) than EHB-1 media when normalized to chlorophyll concentration (0.77 +/- 0.44 micromol H(2) mg Chl(-1) h(-1)) and optical density (1.53 +/- 0.07 nmol H(2) OD(730) (-1) h(-1)), respectively. Both optimization methods have unique benefits and drawbacks that are identified and discussed in this study. (c) 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009.

  18. A Toxin-Antitoxin System VapBC15 from Synechocystis sp. PCC 6803 Shows Distinct Regulatory Features

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    Qian Fei

    2018-03-01

    Full Text Available Type II toxin–antitoxin (TA systems play important roles in bacterial stress survival by regulating cell growth or death. They are highly abundant in cyanobacteria yet remain poorly characterized. Here, we report the identification and regulation of a putative type II TA system from Synechocystis PCC6803, VapBC15. The VapBC15 system is encoded by the chromosomal operon vapBC15. Exogenous expression of VapC15 dramatically arrested cell growth of Escherichia coli and reduced the numbers of colony-forming units (CFU. The VapC15 toxicity could be which was counteracted neutralized by simultaneous or delayed production of VapB15. Biochemical analysis demonstrated the formation of VapB15-VapC15 complexes by the physical interaction between VapB15 and VapC15. Notably, the VapB15 antitoxin up-regulated the transcription of the vapBC15 operon by directly binding to the promoter region, and the VapC15 toxin abolished the up-regulatory effect by destabilizing the binding. Moreover, VapB15 can be degraded by the proteases Lons and ClpXP2s from Synechocystis PCC6803, thus activating the latent toxicity of VapBC15. These findings suggest that VapBC15 represents a genuine TA system that utilizes a distinct mechanism to regulate toxin activity.

  19. Production of 10S-hydroxy-8(E)-octadecenoic acid from oleic acid by whole recombinant Escherichia coli cells expressing 10S-dioxygenase from Nostoc punctiforme PCC 73102 with the aid of a chaperone.

    Science.gov (United States)

    Kim, Min-Ji; Seo, Min-Ju; Shin, Kyung-Chul; Oh, Deok-Kun

    2017-01-01

    To increase the production of 10S-hydroxy-8(E)-octadecenoic acid from oleic acid by whole recombinant Escherichia coli cells expressing Nostoc punctiforme 10S-dioxygenase with the aid of a chaperone. The optimal conditions for 10S-hydroxy-8(E)-octadecenoic acid production by recombinant cells co-expressing chaperone plasmid were pH 9, 35 °C, 15 % (v/v) dimethyl sulfoxide, 40 g cells l -1 , and 10 g oleic acid l -1 . Under these conditions, recombinant cells co-expressing chaperone plasmid produced 7.2 g 10S-hydroxy-8(E)-octadecenoic acid l -1 within 30 min, with a conversion yield of 72 % (w/w) and a volumetric productivity of 14.4 g l -1 h -1 . The activity of recombinant cells expressing 10S-dioxygenase was increased by 200 % with the aid of a chaperone, demonstrating the first biotechnological production of 10S-hydroxy-8(E)-octadecenoic acid using recombinant cells expressing 10S-dioxygenase.

  20. The Combined Use of in Silico, in Vitro, and in Vivo Analyses to Assess Anti-cancerous Potential of a Bioactive Compound from Cyanobacterium Nostoc sp. MGL001

    Directory of Open Access Journals (Sweden)

    Niveshika

    2017-11-01

    Full Text Available Escalating incidences of cancer, especially in developed and developing countries, demand evaluation of potential unexplored natural drug resources. Here, anticancer potential of 9-Ethyliminomethyl-12-(morpholin-4-ylmethoxy-5,8,13,16-tetraaza -hexacene-2,3-dicarboxylic acid (EMTAHDCA isolated from fresh water cyanobacterium Nostoc sp. MGL001 was screened through in silico, in vitro, and in vivo studies. For in silico analysis, EMTAHDCA was selected as ligand and 11 cancer related proteins (Protein Data Bank ID: 1BIX, 1NOW, 1TE6, 2RCW, 2UVL, 2VCJ, 3CRY, 3HQU, 3NMQ, 5P21, and 4B7P which are common targets of various anticancer drugs were selected as receptors. The results obtained from in silico analysis showed that EMTAHDCA has strong binding affinity for all the 11 target protein receptors. The ability of EMTAHDCA to bind active sites of cancer protein targets indicated that it is functionally similar to commercially available anticancer drugs. For assessing cellular metabolic activities, in vitro studies were performed by using calorimetric assay viz. 3-(4,5-dimethylthiazol-2-yl-2,5 diphenyltetrazolium bromide (MTT. Results showed that EMTAHDCA induced significant cytotoxic response against Dalton's lymphoma ascites (DLA cells in a dose and time dependent manner with an inhibitory concentration (IC50 value of 372.4 ng/mL after 24 h of incubation. However, in case of normal bone marrow cells, the EMTAHDCA did not induce cytotoxicity as the IC50 value was not obtained even with higher dose of 1,000 ng/mL EMTAHDCA. Further, in vivo studies revealed that the median life span/survival days of tumor bearing mice treated with EMTAHDCA increased significantly with a fold change of ~1.9 and 1.81 corresponding to doses of 5 and 10 mg/kg body weight (B.W. of EMTAHDCA respectively, as compared to the DL group. Our results suggest that 5 mg/kg B.W. is effective since the dose of 10 mg/kg B.W. did not show any significant difference as compared to 5 mg/kg B

  1. Looking at the stability of life-support microorganisms in space : the MELGEN activity highlights the cyanobacterium Arthrospira sp. PCC8005

    Science.gov (United States)

    Morin, Nicolas

    The MELGEN activity (MELiSSA Genetic Stability Study) mainly covers the molecular aspects of the regenerative life-support system MELiSSA (Micro-Ecological Life Support System Alternative) of the European Space Agency (ESA). The general objective of MELGEN is to establish and validate methods and the related hardware in order to detect genetic instability and microbial contaminants in the MELISSA compartments. This includes (1) a genetic description of the MELISSA strains, (2) studies of microbial behavior and genetic stability in bioreactors and (3) the detection of chemical, genetical and biological contamination and their effect on microbial metabolism. Selected as oxygen producer and complementary food source, the cyanobacterium Arthrospira sp. PCC8005 plays a major role within the MELiSSA loop. As the genomic information on this organism was insufficient, sequencing of its genome was proposed at the French National Sequencing Center, Genoscope, as a joint effort between ESA and different laboratories. So far, a preliminary assembly of 16 contigs representing circa 6.3 million basepairs was obtained. Even though the finishing of the genome is on its way, automatic annotation of the contigs has already been performed on the MaGe annotation platform, and curation of the sequence is currently being carried out, with a special focus on biosynthesis pathways, photosynthesis, and maintenance processes of the cell. According to the index of repetitiveness described by Haubold and Wiehe (2006), we discovered that the genome of Arthrospira sp. is among the 50 most repeated bacterial genomes sequenced to date. Thanks to the sequencing project, we have identified and catalogued mobile genetics elements (MGEs) dispersed throughout the unique chromosome of this cyanobacterium. They represent a quite large proportion of the genome, as genes identified as putative transposases are indeed found in circa 5 Results : We currently have a first draft of the complete genome of

  2. Alterations in protein synthesis in the cyanobacterium Synechococcus sp. Strain PCC 6301 in response to Calendula Micrantha extract with the Molluscicidal activity

    International Nuclear Information System (INIS)

    Hammouda, O.H.E.; Borbely, G.

    1995-01-01

    The response to the extract of the egyptian wild herb Calendula Micrantha, with the Molluscicidal activity, was examined in the unicellular no bacterium Synechococcus sp. strain PCC 6301. growth and chlorophyll of the cells were only slightly affected by low plant extract concentrations but were drastically reduced by high concentration. the rate of protein synthesis progressively decreased by increasing extract concentration. the cells preferentially induced the synthesis of a limited number of polypeptides in response to the treatment. Among the induced polypeptides were those with apparent molecular weights of 161 K (161.000), 96.7 K, 93.4 K, 69.9 K, 59 K, 49 K, 45 K, 35 K, 32.4 K, 28 K, 24 K, 21.7 K, 18 K and 16 K based on their mobilities in gel electrophoresis. these initial studies suggest that the plant extract exerted certain stress which stimulated alteration in the pattern of protein synthesis in Synechococcus sp. some of induced polypeptides are similar to that known to occur in other stresses specially heat shock stress. 3 figs

  3. Alterations in protein synthesis in the cyanobacterium Synechococcus sp. Strain PCC 6301 in response to Calendula Micrantha extract with the Molluscicidal activity.

    Energy Technology Data Exchange (ETDEWEB)

    Hammouda, O H.E. [Botany Department, Faculty of Science, Cairo University, Beni-Suef branch. Beni Suef (Egypt); Borbely, G [Institute of plant physiology, Biological Research Center, Szeged H-6701, (Hungary)

    1995-10-01

    The response to the extract of the egyptian wild herb Calendula Micrantha, with the Molluscicidal activity, was examined in the unicellular no bacterium Synechococcus sp. strain PCC 6301. growth and chlorophyll of the cells were only slightly affected by low plant extract concentrations but were drastically reduced by high concentration. the rate of protein synthesis progressively decreased by increasing extract concentration. the cells preferentially induced the synthesis of a limited number of polypeptides in response to the treatment. Among the induced polypeptides were those with apparent molecular weights of 161 K (161.000), 96.7 K, 93.4 K, 69.9 K, 59 K, 49 K, 45 K, 35 K, 32.4 K, 28 K, 24 K, 21.7 K, 18 K and 16 K based on their mobilities in gel electrophoresis. these initial studies suggest that the plant extract exerted certain stress which stimulated alteration in the pattern of protein synthesis in Synechococcus sp. some of induced polypeptides are similar to that known to occur in other stresses specially heat shock stress. 3 figs.

  4. Quantitative iTRAQ LC-MS/MS proteomics reveals metabolic responses to biofuel ethanol in cyanobacterial Synechocystis sp. PCC 6803.

    Science.gov (United States)

    Qiao, Jianjun; Wang, Jiangxin; Chen, Lei; Tian, Xiaoxu; Huang, Siqiang; Ren, Xiaoyue; Zhang, Weiwen

    2012-11-02

    Recent progress in metabolic engineering has led to autotrophic production of ethanol in various cyanobacterial hosts. However, cyanobacteria are known to be sensitive to ethanol, which restricts further efforts to increase ethanol production levels in these renewable host systems. To understand the mechanisms of ethanol tolerance so that engineering more robust cyanobacterial hosts can be possible, in this study, the responses of model cyanobacterial Synechocystis sp. PCC 6803 to ethanol were determined using a quantitative proteomics approach with iTRAQ LC-MS/MS technologies. The resulting high-quality proteomic data set consisted of 24,887 unique peptides corresponding to 1509 identified proteins, a coverage of approximately 42% of the predicted proteins in the Synechocystis genome. Using a cutoff of 1.5-fold change and a p-value less than 0.05, 135 and 293 unique proteins with differential abundance levels were identified between control and ethanol-treated samples at 24 and 48 h, respectively. Functional analysis showed that the Synechocystis cells employed a combination of induced common stress response, modifications of cell membrane and envelope, and induction of multiple transporters and cell mobility-related proteins as protection mechanisms against ethanol toxicity. Interestingly, our proteomic analysis revealed that proteins related to multiple aspects of photosynthesis were up-regulated in the ethanol-treated Synechocystis cells, consistent with increased chlorophyll a concentration in the cells upon ethanol exposure. The study provided the first comprehensive view of the complicated molecular mechanisms against ethanol stress and also provided a list of potential gene targets for further engineering ethanol tolerance in Synechocystis PCC 6803.

  5. Viability of dried filaments, survivability and reproduction under water stress, and survivability following heat and UV exposure in Lyngbya martensiana, Oscillatoria agardhii, Nostoc calcicola, Hormidium fluitans, Spirogyra sp. and Vaucheria geminata

    International Nuclear Information System (INIS)

    Agrawal, S.C.; Singh, V.

    2002-01-01

    The aim of our study was to determine how long and to what extent Lyngbya martensiana, Oscillatoria agardhii, Nostoc calcicola, Hormidium fluitans and Vaucheria geminata tolerate dry storage at different temperatures, UV-light radiation and water stress imposed by growing them on media with a high agar content and/or in NaCl-containing liquid media. Dried vegetative filaments of Spirogyra sp., Vaucheria geminata and Nostoc calcicola died within 0,5, 1 and 4 h, respectively; those of Hormidium fluitans, Oscillatoria agardhii and Lyngbya martensiana retained viability for 3, 5 and 10 d, respectively. L. martensiana and O. agardhii tolerated 0.8 mol/L NaCl. The resistance to desiccation in L. martensiana and O. agardhii exhibited similar dependence as that to frost, to heat and UV light. The water stress imposed on growing algae either on high-agar solid media or in NaCl-containing liquid media reduced hormogonium formation in L. martensiana and O. agardhii; hetero-cyst and akinete formation in N. calcicola and fragmentation in H. fluitans. In all studied algae the stress reduced at various levels the survival of vegetative parts. Generally, algal body form and composition rather than habitats seem to decide primarily the level of resistance against various stress conditions

  6. Photosynthetic versatility in the genome of Geitlerinema sp. PCC 9228 (formerly Oscillatoria limnetica ‘Solar Lake’, a model anoxygenic photosynthetic cyanobacterium

    Directory of Open Access Journals (Sweden)

    Sharon L Grim

    2016-10-01

    Full Text Available Anoxygenic cyanobacteria that use sulfide as the electron donor for photosynthesis are a potentially influential but poorly constrained force on Earth’s biogeochemistry. Their versatile metabolism may have boosted primary production and nitrogen cycling in euxinic coastal margins in the Proterozoic. In addition, they represent a biological mechanism for limiting the accumulation of atmospheric oxygen, especially before the Great Oxidation Event and in the low-oxygen conditions of the Proterozoic. In this study, we describe the draft genome sequence of Geitlerinema sp. PCC 9228, formerly Oscillatoria limnetica ‘Solar Lake’, a mat-forming diazotrophic cyanobacterium that can switch between oxygenic photosynthesis and sulfide-based anoxygenic photosynthesis. Geitlerinema possesses three variants of psbA, which encodes protein D1, a core component of the photosystem II reaction center. Phylogenetic analyses indicate that one variant is closely affiliated with cyanobacterial psbA genes that code for a D1 protein used for oxygen-sensitive processes. Another version is phylogenetically similar to cyanobacterial psbA genes that encode D1 proteins used under microaerobic conditions, and the third variant may be cued to high light and/or elevated oxygen concentrations. Geitlerinema has the canonical gene for sulfide quinone reductase (SQR used in cyanobacterial anoxygenic photosynthesis and a putative transcriptional regulatory gene in the same operon. Another operon with a second, distinct sqr and regulatory gene is present, and is phylogenetically related to sqr genes used for high sulfide concentrations. The genome has a comprehensive nif gene suite for nitrogen fixation, supporting previous observations of nitrogenase activity. Geitlerinema possesses a bidirectional hydrogenase rather than the uptake hydrogenase typically used by cyanobacteria in diazotrophy. Overall, the genome sequence of Geitlerinema sp. PCC 9228 highlights potential

  7. Photosynthetic Versatility in the Genome of Geitlerinema sp. PCC 9228 (Formerly Oscillatoria limnetica ‘Solar Lake’), a Model Anoxygenic Photosynthetic Cyanobacterium

    Science.gov (United States)

    Grim, Sharon L.; Dick, Gregory J.

    2016-01-01

    Anoxygenic cyanobacteria that use sulfide as the electron donor for photosynthesis are a potentially influential but poorly constrained force on Earth’s biogeochemistry. Their versatile metabolism may have boosted primary production and nitrogen cycling in euxinic coastal margins in the Proterozoic. In addition, they represent a biological mechanism for limiting the accumulation of atmospheric oxygen, especially before the Great Oxidation Event and in the low-oxygen conditions of the Proterozoic. In this study, we describe the draft genome sequence of Geitlerinema sp. PCC 9228, formerly Oscillatoria limnetica ‘Solar Lake’, a mat-forming diazotrophic cyanobacterium that can switch between oxygenic photosynthesis and sulfide-based anoxygenic photosynthesis (AP). Geitlerinema possesses three variants of psbA, which encodes protein D1, a core component of the photosystem II reaction center. Phylogenetic analyses indicate that one variant is closely affiliated with cyanobacterial psbA genes that code for a D1 protein used for oxygen-sensitive processes. Another version is phylogenetically similar to cyanobacterial psbA genes that encode D1 proteins used under microaerobic conditions, and the third variant may be cued to high light and/or elevated oxygen concentrations. Geitlerinema has the canonical gene for sulfide quinone reductase (SQR) used in cyanobacterial AP and a putative transcriptional regulatory gene in the same operon. Another operon with a second, distinct sqr and regulatory gene is present, and is phylogenetically related to sqr genes used for high sulfide concentrations. The genome has a comprehensive nif gene suite for nitrogen fixation, supporting previous observations of nitrogenase activity. Geitlerinema possesses a bidirectional hydrogenase rather than the uptake hydrogenase typically used by cyanobacteria in diazotrophy. Overall, the genome sequence of Geitlerinema sp. PCC 9228 highlights potential cyanobacterial strategies to cope with

  8. Long-Term Acclimation of the Cyanobacterium Synechocystis sp PCC 6803 to High Light Is Accompanied by an Enhanced Production of Chlorophyll That Is Preferentially Channeled to Trimeric Photosystem I

    Czech Academy of Sciences Publication Activity Database

    Kopečná, Jana; Komenda, Josef; Bučinská, Lenka; Sobotka, Roman

    2012-01-01

    Roč. 160, č. 4 (2012), s. 2239-2250 ISSN 0032-0889 R&D Projects: GA MŠk(CZ) ED2.1.00/03.0110; GA ČR GAP501/10/1000; GA ČR GBP501/12/G055 Institutional support: RVO:61388971 Keywords : CAB-LIKE-PROTEINS * SP PCC-6803 * PHOTOSYNTHETIC APPARATUS Subject RIV: EE - Microbiology, Virology Impact factor: 6.555, year: 2012

  9. Role of phosphatidylglycerol in the function and assembly of Photosystem II reaction center, studied in a cdsA-inactivated PAL mutant strain of Synechocystis sp. PCC6803 that lacks phycobilisomes

    Czech Academy of Sciences Publication Activity Database

    Laczkó-Dobos, H.; Ughy, B.; Tóth, S. Z.; Komenda, Josef; Zsiros, O.; Domonkos, I.; Párducz, A.; Bogos, B.; Komura, M.; Itoh, S.; Gombos, Z.

    2008-01-01

    Roč. 1777, č. 9 (2008), s. 1184-1194 ISSN 0005-2728 R&D Projects: GA AV ČR IAA400200801 Grant - others:HU(HU) OTKA T60109; HU(HU) OTKA T68692 Institutional research plan: CEZ:AV0Z50200510 Keywords : synechocystis sp. pcc6803 * phosphatidylglycerol * photosystem II Subject RIV: EE - Microbiology, Virology Impact factor: 4.447, year: 2008

  10. Transcription activation by NtcA and 2-oxoglutarate of three genes involved in heterocyst differentiation in the cyanobacterium Anabaena sp. strain PCC 7120.

    Science.gov (United States)

    Valladares, Ana; Flores, Enrique; Herrero, Antonia

    2008-09-01

    In Anabaena sp. strain PCC 7120, differentiation of heterocysts takes place in response to the external cue of combined nitrogen deprivation, allowing the organism to fix atmospheric nitrogen in oxic environments. NtcA, a global transcriptional regulator of cyanobacteria, is required for activation of the expression of multiple genes involved in heterocyst differentiation, including key regulators that are specific to the process. We have set up a fully defined in vitro system, which includes the purified Anabaena RNA polymerase, and have studied the effects of NtcA and its signaling effector 2-oxoglutarate on RNA polymerase binding, open complex formation, and transcript production from promoters of the hetC, nrrA, and devB genes that are activated by NtcA at different stages of heterocyst differentiation. Both RNA polymerase and NtcA could specifically bind to the target DNA in the absence of any effector. 2-Oxoglutarate had a moderate positive effect on NtcA binding, and NtcA had a limited positive effect on RNA polymerase recruitment at the promoters. However, a stringent requirement of both NtcA and 2-oxoglutarate was observed for the detection of open complexes and transcript production at the three investigated promoters. These results support a key role for 2-oxoglutarate in transcription activation in the developing heterocyst.

  11. Anchoring a plant cytochrome P450 via PsaM to the thylakoids in Synechococcus sp. PCC 7002: evidence for light-driven biosynthesis.

    Directory of Open Access Journals (Sweden)

    Lærke Münter Lassen

    Full Text Available Plants produce an immense variety of specialized metabolites, many of which are of high value as their bioactive properties make them useful as for instance pharmaceuticals. The compounds are often produced at low levels in the plant, and due to their complex structures, chemical synthesis may not be feasible. Here, we take advantage of the reducing equivalents generated in photosynthesis in developing an approach for producing plant bioactive natural compounds in a photosynthetic microorganism by functionally coupling a biosynthetic enzyme to photosystem I. This enables driving of the enzymatic reactions with electrons extracted from the photosynthetic electron transport chain. As a proof of concept, we have genetically fused the soluble catalytic domain of the cytochrome P450 CYP79A1, originating from the endoplasmic reticulum membranes of Sorghum bicolor, to a photosystem I subunit in the cyanobacterium Synechococcus sp. PCC 7002, thereby targeting it to the thylakoids. The engineered enzyme showed light-driven activity both in vivo and in vitro, demonstrating the possibility to achieve light-driven biosynthesis of high-value plant specialized metabolites in cyanobacteria.

  12. Resistance, accumulation and transformation of selenium by the cyanobacterium Synechocystis sp. PCC 6803 after exposure to inorganic SeVI or SeIV

    International Nuclear Information System (INIS)

    Gouget, B.; Avoscan, L.; Collins, R.; Carriere, M.; Sarret, G.

    2005-01-01

    Our purpose was to investigate the ability of Synechocystis sp. PCC 6803, a photosynthetic prokaryote isolated from fresh water, to resist, incorporate and reduce the oxidized forms of selenium including selenite and selenate, the major selenium species present in aquatic systems. Selenium speciation and the chemical intermediates during selenium transformation were determined by X-ray absorption near edge structure (XANES) spectroscopy. The possible internalisation pathways involving selenium and the metabolic fate of selenate and selenite were examined. Selenate metabolism seemed to proceed via the sulfate reduction pathway resulting in the formation of the R-Se-H, R-Se-R and R-Se-Se-R species. The transformation of selenate to toxic amino acids may explain the high sensitivity of Synechocystis to selenate. Several mechanisms of selenium reduction seem to complete during selenite assimilation. A specific mechanism may transform internalised selenite into selenide and, subsequently induce the biosynthesis of selenoproteins. A non-specific mechanism may interfere with thiols, such as glutathione in the cell cytoplasm, or with proteins in the periplasm of the bacteria, notably thioredoxins. Several hypotheses concerning the complex transformation of selenium in Synechocystis could therefore be proposed. (orig.)

  13. Cloning expression and analysis of phytochelatin synthase (pcs) gene from Anabaena sp. PCC 7120 offering multiple stress tolerance in Escherichia coli

    International Nuclear Information System (INIS)

    Chaurasia, Neha; Mishra, Yogesh; Rai, Lal Chand

    2008-01-01

    Phytochelatin synthase (PCS) is involved in the synthesis of phytochelatins (PCs), plays role in heavy metal detoxification. The present study describes for first time the functional expression and characterization of pcs gene of Anabaena sp. PCC 7120 in Escherichia coli in terms of offering protection against heat, salt, carbofuron (pesticide), cadmium, copper, and UV-B stress. The involvement of pcs gene in tolerance to above abiotic stresses was investigated by cloning of pcs gene in expression vector pGEX-5X-2 and its transformation in E. coli BL21 (DE3). The E. coli cells transformed with pGEX-5X-pcs showed better growth than control cells (pGEX-5X-2) under temperature (47 deg. C), NaCl (6% w/v), carbofuron (0.025 mg ml -1 ), CdCl 2 (4 mM), CuCl 2 (1 mM), and UV-B (10 min) exposure. The enhanced expression of pcs gene revealed by RT-PCR analysis under above stresses at different time intervals further advocates its role in tolerance against above abiotic stresses

  14. Disruption of the ndhF1 gene affects Chl fluorescence through state transition in the Cyanobacterium Synechocystis sp. PCC 6803, resulting in apparent high efficiency of photosynthesis.

    Science.gov (United States)

    Ogawa, Takako; Harada, Tetsuyuki; Ozaki, Hiroshi; Sonoike, Kintake

    2013-07-01

    In Synechocystis sp. PCC 6803, the disruption of the ndhF1 gene (slr0844), which encodes a subunit of one of the NDH-1 complexes (NDH-1L complex) serving for respiratory electron transfer, causes the largest change in Chl fluorescence induction kinetics among the kinetics of 750 disruptants searched in the Fluorome, the cyanobacterial Chl fluorescence database. The cause of the explicit phenotype of the ndhF1 disruptant was examined by measurements of the photosynthetic rate, Chl fluorescence and state transition. The results demonstrate that the defects in respiratory electron transfer obviously have great impact on Chl fluorescence in cyanobacteria. The inactivation of NDH-1L complexes involving electron transfer from NDH-1 to plastoquinone (PQ) would result in the oxidation of the PQ pool, leading to the transition to State 1, where the yield of Chl fluorescence is high. Apparently, respiration, although its rate is far lower than that of photosynthesis, could affect Chl fluorescence through the state transition as leverage. The disruption of the ndhF1 gene caused lower oxygen-evolving activity but the estimated electron transport rate from Chl fluorescence measurements was faster in the mutant than in the wild-type cells. The discrepancy could be ascribed to the decreased level of non-photochemical quenching due to state transition. One must be cautious when using the Chl fluorescence parameter to estimate photosynthesis in mutants defective in state transition.

  15. RNA-seq based identification and mutant validation of gene targets related to ethanol resistance in cyanobacterial Synechocystis sp. PCC 6803

    Directory of Open Access Journals (Sweden)

    Wang Jiangxin

    2012-12-01

    Full Text Available Abstract Background Fermentation production of biofuel ethanol consumes agricultural crops, which will compete directly with the food supply. As an alternative, photosynthetic cyanobacteria have been proposed as microbial factories to produce ethanol directly from solar energy and CO2. However, the ethanol productivity from photoautotrophic cyanobacteria is still very low, mostly due to the low tolerance of cyanobacterial systems to ethanol stress. Results To build a foundation necessary to engineer robust ethanol-producing cyanobacterial hosts, in this study we applied a quantitative transcriptomics approach with a next-generation sequencing technology, combined with quantitative reverse-transcript PCR (RT-PCR analysis, to reveal the global metabolic responses to ethanol in model cyanobacterial Synechocystis sp. PCC 6803. The results showed that ethanol exposure induced genes involved in common stress responses, transporting and cell envelope modification. In addition, the cells can also utilize enhanced polyhydroxyalkanoates (PHA accumulation and glyoxalase detoxication pathway as means against ethanol stress. The up-regulation of photosynthesis by ethanol was also further confirmed at transcriptional level. Finally, we used gene knockout strains to validate the potential target genes related to ethanol tolerance. Conclusion RNA-Seq based global transcriptomic analysis provided a comprehensive view of cellular response to ethanol exposure. The analysis provided a list of gene targets for engineering ethanol tolerance in cyanobacterium Synechocystis.

  16. (1)H, (13)C, (15)N backbone and side-chain resonance assignment of Nostoc sp. C139A variant of the heme-nitric oxide/oxygen binding (H-NOX) domain.

    Science.gov (United States)

    Alexandropoulos, Ioannis I; Argyriou, Aikaterini I; Marousis, Kostas D; Topouzis, Stavros; Papapetropoulos, Andreas; Spyroulias, Georgios A

    2016-10-01

    The H-NOX (Heme-nitric oxide/oxygen binding) domain is conserved across eukaryotes and bacteria. In human soluble guanylyl cyclase (sGC) the H-NOX domain functions as a sensor for the gaseous signaling agent nitric oxide (NO). sGC contains the heme-binding H-NOX domain at its N-terminus, which regulates the catalytic site contained within the C-terminal end of the enzyme catalyzing the conversion of GTP (guanosine 5'-triphosphate) to GMP (guanylyl monophosphate). Here, we present the backbone and side-chain assignments of the (1)H, (13)C and (15)N resonances of the 183-residue H-NOX domain from Nostoc sp. through solution NMR.

  17. Transcription profiling of the model cyanobacterium Synechococcus sp. strain PCC 7002 by NextGen (SOLiD™ Sequencing of cDNA

    Directory of Open Access Journals (Sweden)

    Marcus eLudwig

    2011-03-01

    Full Text Available The genome of the unicellular, euryhaline cyanobacterium Synechococcus sp. PCC 7002 encodes about 3200 proteins. Transcripts were detected for nearly all annotated open reading frames by a global transcriptomic analysis by Next-Generation (SOLiDTM sequencing of cDNA. In the cDNA samples sequenced, ~90% of the mapped sequences were derived from the 16S and 23S ribosomal RNAs and ~10% of the sequences were derived from mRNAs. In cells grown photoautotrophically under standard conditions (38 °C, 1% (v/v CO2 in air, 250 µmol photons m-2 s-1, the highest transcript levels (up to 2% of the total mRNA for the most abundantly transcribed genes (e. g., cpcAB, psbA, psaA were generally derived from genes encoding structural components of the photosynthetic apparatus. High light exposure for one hour caused changes in transcript levels for genes encoding proteins of the photosynthetic apparatus, Type-1 NADH dehydrogenase complex and ATP synthase, whereas dark incubation for one hour resulted in a global decrease in transcript levels for photosynthesis-related genes and an increase in transcript levels for genes involved in carbohydrate degradation. Transcript levels for pyruvate kinase and the pyruvate dehydrogenase complex decreased sharply in cells incubated in the dark. Under dark anoxic (fermentative conditions, transcript changes indicated a global decrease in transcripts for respiratory proteins and suggested that cells employ an alternative phosphoenolpyruvate degradation pathway via phosphoenolpyruvate synthase (ppsA and the pyruvate:ferredoxin oxidoreductase (nifJ. Finally, the data suggested that an apparent operon involved in tetrapyrrole biosynthesis and fatty acid desaturation, acsF2-ho2-hemN2-desF, may be regulated by oxygen concentration.

  18. Acetylome analysis reveals the involvement of lysine acetylation in photosynthesis and carbon metabolism in the model cyanobacterium Synechocystis sp. PCC 6803.

    Science.gov (United States)

    Mo, Ran; Yang, Mingkun; Chen, Zhuo; Cheng, Zhongyi; Yi, Xingling; Li, Chongyang; He, Chenliu; Xiong, Qian; Chen, Hui; Wang, Qiang; Ge, Feng

    2015-02-06

    Cyanobacteria are the oldest known life form inhabiting Earth and the only prokaryotes capable of performing oxygenic photosynthesis. Synechocystis sp. PCC 6803 (Synechocystis) is a model cyanobacterium used extensively in research on photosynthesis and environmental adaptation. Posttranslational protein modification by lysine acetylation plays a critical regulatory role in both eukaryotes and prokaryotes; however, its extent and function in cyanobacteria remain unexplored. Herein, we performed a global acetylome analysis on Synechocystis through peptide prefractionation, antibody enrichment, and high accuracy LC-MS/MS analysis; identified 776 acetylation sites on 513 acetylated proteins; and functionally categorized them into an interaction map showing their involvement in various biological processes. Consistent with previous reports, a large fraction of the acetylation sites are present on proteins involved in cellular metabolism. Interestingly, for the first time, many proteins involved in photosynthesis, including the subunits of phycocyanin (CpcA, CpcB, CpcC, and CpcG) and allophycocyanin (ApcA, ApcB, ApcD, ApcE, and ApcF), were found to be lysine acetylated, suggesting that lysine acetylation may play regulatory roles in the photosynthesis process. Six identified acetylated proteins associated with photosynthesis and carbon metabolism were further validated by immunoprecipitation and Western blotting. Our data provide the first global survey of lysine acetylation in cyanobacteria and reveal previously unappreciated roles of lysine acetylation in the regulation of photosynthesis. The provided data set may serve as an important resource for the functional analysis of lysine acetylation in cyanobacteria and facilitate the elucidation of the entire metabolic networks and photosynthesis process in this model cyanobacterium.

  19. Trade-Off between Growth and Carbohydrate Accumulation in Nutrient-Limited Arthrospira sp. PCC 8005 Studied by Integrating Transcriptomic and Proteomic Approaches.

    Directory of Open Access Journals (Sweden)

    Orily Depraetere

    Full Text Available Cyanobacteria have a strong potential for biofuel production due to their ability to accumulate large amounts of carbohydrates. Nitrogen (N stress can be used to increase the content of carbohydrates in the biomass, but it is expected to reduce biomass productivity. To study this trade-off between carbohydrate accumulation and biomass productivity, we characterized the biomass productivity, biomass composition as well as the transcriptome and proteome of the cyanobacterium Arthrospira sp. PCC 8005 cultured under N-limiting and N-replete conditions. N limitation resulted in a large increase in the carbohydrate content of the biomass (from 14 to 74% and a decrease in the protein content (from 37 to 10%. Analyses of fatty acids indicated that no lipids were accumulated under N-limited conditions. Nevertheless, it did not affect the biomass productivity of the culture up to five days after N was depleted from the culture medium. Transcriptomic and proteomic analysis indicated that de novo protein synthesis was down-regulated in the N-limited culture. Proteins were degraded and partly converted into carbohydrates through gluconeogenesis. Cellular N derived from protein degradation was recycled through the TCA and GS-GOGAT cycles. In addition, photosynthetic energy production and carbon fixation were both down-regulated, while glycogen synthesis was up-regulated. Our results suggested that N limitation resulted in a redirection of photosynthetic energy from protein synthesis to glycogen synthesis. The fact that glycogen synthesis has a lower energy demand than protein synthesis might explain why Arthrospira is able to achieve a similar biomass productivity under N-limited as under N-replete conditions despite the fact that photosynthetic energy production was impaired by N limitation.

  20. Finding novel relationships with integrated gene-gene association network analysis of Synechocystis sp. PCC 6803 using species-independent text-mining.

    Science.gov (United States)

    Kreula, Sanna M; Kaewphan, Suwisa; Ginter, Filip; Jones, Patrik R

    2018-01-01

    The increasing move towards open access full-text scientific literature enhances our ability to utilize advanced text-mining methods to construct information-rich networks that no human will be able to grasp simply from 'reading the literature'. The utility of text-mining for well-studied species is obvious though the utility for less studied species, or those with no prior track-record at all, is not clear. Here we present a concept for how advanced text-mining can be used to create information-rich networks even for less well studied species and apply it to generate an open-access gene-gene association network resource for Synechocystis sp. PCC 6803, a representative model organism for cyanobacteria and first case-study for the methodology. By merging the text-mining network with networks generated from species-specific experimental data, network integration was used to enhance the accuracy of predicting novel interactions that are biologically relevant. A rule-based algorithm (filter) was constructed in order to automate the search for novel candidate genes with a high degree of likely association to known target genes by (1) ignoring established relationships from the existing literature, as they are already 'known', and (2) demanding multiple independent evidences for every novel and potentially relevant relationship. Using selected case studies, we demonstrate the utility of the network resource and filter to ( i ) discover novel candidate associations between different genes or proteins in the network, and ( ii ) rapidly evaluate the potential role of any one particular gene or protein. The full network is provided as an open-source resource.

  1. Arabinogalactan proteins occur in the free-living cyanobacterium genus Nostoc and in plant-Nostoc symbioses.

    Science.gov (United States)

    Jackson, Owen; Taylor, Oliver; Adams, David G; Knox, J Paul

    2012-10-01

    Arabinogalactan proteins (AGP) are a diverse family of proteoglycans associated with the cell surfaces of plants. AGP have been implicated in a wide variety of plant cell processes, including signaling in symbioses. This study investigates the existence of putative AGP in free-living cyanobacterial cultures of the nitrogen-fixing, filamentous cyanobacteria Nostoc punctiforme and Nostoc sp. strain LBG1 and at the symbiotic interface in the symbioses between Nostoc spp. and two host plants, the angiosperm Gunnera manicata (in which the cyanobacterium is intracellular) and the liverwort Blasia pusilla (in which the cyanobacterium is extracellular). Enzyme-linked immunosorbent assay, immunoblotting, and immunofluorescence analyses demonstrated that three AGP glycan epitopes (recognized by monoclonal antibodies LM14, MAC207, and LM2) are present in free-living Nostoc cyanobacterial species. The same three AGP glycan epitopes are present at the Gunnera-Nostoc symbiotic interface and the LM2 epitope is detected during the establishment of the Blasia-Nostoc symbiosis. Bioinformatic analysis of the N. punctiforme genome identified five putative AGP core proteins that are representative of AGP classes found in plants. These results suggest a possible involvement of AGP in cyanobacterial-plant symbioses and are also suggestive of a cyanobacterial origin of AGP.

  2. Characterization of a microcystin and detection of microcystin synthetase genes from a Brazilian isolate of Nostoc.

    Science.gov (United States)

    Genuário, Diego Bonaldo; Silva-Stenico, Maria Estela; Welker, Martin; Beraldo Moraes, Luiz Alberto; Fiore, Marli Fátima

    2010-04-01

    A nostocalean nitrogen-fixing cyanobacterium isolated from an eutrophic freshwater reservoir located in Piracicaba, São Paulo, Brazil, was evaluated for the production of hepatotoxic cyclic heptapeptides, microcystins. Morphologically this new cyanobacterium strain appears closest to Nostoc, however, in the phylogenetic analysis of 16S rRNA gene it falls into a highly stable cluster distantly only related to the typical Nostoc cluster. Extracts of Nostoc sp. CENA88 cultured cells, investigated using ELISA assay, gave positive results and the microcystin profile revealed by ESI-Q-TOF/MS/MS analysis confirmed the production of [Dha(7)]MCYST-YR. Further, Nostoc sp. CENA88 genomic DNA was analyzed by PCR for sequences of mcyD, mcyE and mcyG genes of microcystin synthetase (mcy) cluster. The result revealed the presence of mcyD, mcyE and mcyG genes with similarities to those from mcy of Nostoc sp. strains 152 and IO-102-I and other cyanobacterial genera. The phylogenetic tree based on concatenated McyG, McyD and McyE amino acids clustered the sequences according to cyanobacterial genera, with exception of the Nostoc sp. CENA88 sequence, which was placed in a clade distantly related from other Nostoc strains, as previously observed also in the 16S rRNA phylogenetic analysis. The present study describes for the first time a Brazilian Nostoc microcystin producer and also the occurrence of demethyl MCYST-YR variant in this genus. The sequenced Nostoc genes involved in the microcystin synthesis can contribute to a better understanding of the toxigenicity and evolution of this cyanotoxin. Copyright 2009 Elsevier Ltd. All rights reserved.

  3. Produção de exopolissacarídeos pela cianobactéria Nostoc sp em diferentes concentrações de nitrogênio e glicose

    Directory of Open Access Journals (Sweden)

    Liege Abdallah Kawai

    2006-05-01

    Full Text Available Cianobactérias são microrganismos procariontes que, durante o crescimento celular, são capazes de produzir exopolissacarídeos (EPS. Devido à diversidade bioquímica destes, podem ser excelentes para vários fins biotecnológicos, tendo aplicações em indústrias alimentícias, têxteis, de tintas, cosméticos, de papel, e farmacêuticas, como floculantes, espessantes ou estabilizadores, substituindo os polissacarídeos de macroalgas e plantas. Além disso, as cianobactérias apresentam taxas maiores de crescimento e são mais fáceis de manipular do que plantas e macroalgas. Este estudo teve por objetivo otimizar a produção de EPS no meio BG11, com relação a diferentes concentrações de nitrogênio e glicose do meio de cultivo na produção de EPS e biomassa pela cianobactéria Nostoc sp.

  4. Biochemical characterization of sunscreening mycosporine-like amino acids from two Nostoc species inhabiting diverse habitats.

    Science.gov (United States)

    Richa; Sinha, Rajeshwar P

    2015-01-01

    We have screened two Nostoc species inhabiting diverse habitats for the presence of sunscreening mycosporine-like amino acid (MAA) compounds. The identification and characterization of one MAA (RT 3.1-3.8 min, λmax -334 nm) from both Nostoc species were performed using absorption spectroscopy, high-performance liquid chromatography (HPLC), electrospray ionization-mass spectrometry (ESI-MS), Fourier transform infrared (FTIR) spectroscopy and nuclear magnetic resonance (NMR) spectroscopy. Shinorine and porphyra-334 were commonly present in both Nostoc sp. strain HKAR-2 and Nostoc sp. strain HKAR-6. Nostoc sp. strain HKAR-2 also showed the presence of an unknown MAAs with retention time of 6.9 min and a corresponding λmax of 334 nm. Present investigation clearly demonstrated the presence of diverse profile of MAAs in the hot spring cyanobacterium in comparison to the rice field isolate. Thus, Nostoc sp. strain HKAR-2 would be a better source for the production of MAAs that can be used as a potent natural sunscreen against UV-B irradiation.

  5. Bioinformatic evaluation of L-arginine catabolic pathways in 24 cyanobacteria and transcriptional analysis of genes encoding enzymes of L-arginine catabolism in the cyanobacterium Synechocystis sp. PCC 6803

    Directory of Open Access Journals (Sweden)

    Pistorius Elfriede K

    2007-11-01

    Full Text Available Abstract Background So far very limited knowledge exists on L-arginine catabolism in cyanobacteria, although six major L-arginine-degrading pathways have been described for prokaryotes. Thus, we have performed a bioinformatic analysis of possible L-arginine-degrading pathways in cyanobacteria. Further, we chose Synechocystis sp. PCC 6803 for a more detailed bioinformatic analysis and for validation of the bioinformatic predictions on L-arginine catabolism with a transcript analysis. Results We have evaluated 24 cyanobacterial genomes of freshwater or marine strains for the presence of putative L-arginine-degrading enzymes. We identified an L-arginine decarboxylase pathway in all 24 strains. In addition, cyanobacteria have one or two further pathways representing either an arginase pathway or L-arginine deiminase pathway or an L-arginine oxidase/dehydrogenase pathway. An L-arginine amidinotransferase pathway as a major L-arginine-degrading pathway is not likely but can not be entirely excluded. A rather unusual finding was that the cyanobacterial L-arginine deiminases are substantially larger than the enzymes in non-photosynthetic bacteria and that they are membrane-bound. A more detailed bioinformatic analysis of Synechocystis sp. PCC 6803 revealed that three different L-arginine-degrading pathways may in principle be functional in this cyanobacterium. These are (i an L-arginine decarboxylase pathway, (ii an L-arginine deiminase pathway, and (iii an L-arginine oxidase/dehydrogenase pathway. A transcript analysis of cells grown either with nitrate or L-arginine as sole N-source and with an illumination of 50 μmol photons m-2 s-1 showed that the transcripts for the first enzyme(s of all three pathways were present, but that the transcript levels for the L-arginine deiminase and the L-arginine oxidase/dehydrogenase were substantially higher than that of the three isoenzymes of L-arginine decarboxylase. Conclusion The evaluation of 24

  6. Selective Production of 9R-Hydroxy-10E,12Z,15Z-Octadecatrienoic Acid from α-Linolenic Acid in Perilla Seed Oil Hydrolyzate by a Lipoxygenase from Nostoc Sp. SAG 25.82.

    Science.gov (United States)

    Kim, Kyoung-Rok; An, Jung-Ung; Lee, Seon-Hwa; Oh, Deok-Kun

    2015-01-01

    Hydroxy fatty acids (HFAs) derived from omega-3 polyunsaturated fatty acids have been known as versatile bioactive molecules. However, its practical production from omega-3 or omega-3 rich oil has not been well established. In the present study, the stereo-selective enzymatic production of 9R-hydroxy-10E,12Z,15Z-octadecatrienoic acid (9R-HOTE) from α-linolenic acid (ALA) in perilla seed oil (PO) hydrolyzate was achieved using purified recombinant 9R-lipoxygenase (9R-LOX) from Nostoc sp. SAG 25.82. The specific activity of the enzyme followed the order linoleic acid (LA) > ALA > γ-linolenic acid (GLA). A total of 75% fatty acids (ALA and LA) were used as a substrate for 9R-LOX from commercial PO by hydrolysis of Candida rugosa lipase. The optimal reaction conditions for the production of 9R-HOTE from ALA using 9R-LOX were pH 8.5, 15°C, 5% (v/v) acetone, 0.2% (w/v) Tween 80, 40 g/L ALA, and 1 g/L enzyme. Under these conditions, 9R-LOX produced 37.6 g/L 9R-HOTE from 40 g/L ALA for 1 h, with a conversion yield of 94% and a productivity of 37.6 g/L/h; and the enzyme produced 34 g/L 9R-HOTE from 40 g/L ALA in PO hydrolyzate for 1 h, with a conversion yields of 85% and a productivity of 34 g/L/h. The enzyme also converted 9R-hydroxy-10E,12Z-octadecadienoic acid (9R-HODE) from 40 g/L LA for 1.0 h, with a conversion yield of 95% and a productivity of 38.4 g/L. This is the highest productivity of HFA from both ALA and ALA-rich vegetable oil using LOX ever reported. Therefore, our result suggests an efficient method for the production of 9R-HFAs from LA and ALA in vegetable oil using recombinant LOX in biotechnology.

  7. DNA Probes Show Genetic Variation in Cyanobacterial Symbionts of the Azolla Fern and a Closer Relationship to Free-Living Nostoc Strains than to Free-Living Anabaena Strains

    Science.gov (United States)

    Plazinski, Jacek; Zheng, Qi; Taylor, Rona; Croft, Lynn; Rolfe, Barry G.; Gunning, Brian E. S.

    1990-01-01

    Twenty-two isolates of Anabaena azollae derived from seven Azolla species from various geographic and ecological sources were characterized by DNA-DNA hybridization. Cloned DNA fragments derived from the genomic sequences of three different A. azollae isolates were used to detect restriction fragment length polymorphism among all symbiotic anabaenas. DNA clones were radiolabeled and hybridized against southern blot transfers of genomic DNAs of different isolates of A. azollae digested with restriction endonucleases. Eight DNA probes were selected to identify the Anabaena strains tested. Two were strain specific and hybridized only to A. azollae strains isolated from Azolla microphylla or Azolla caroliniana. One DNA probe was section specific (hybridized only to anabaenas isolated from Azolla ferns representing the section Euazolla), and five other probes gave finer discrimination among anabaenas representing various ecotypes of Azolla species. These cloned genomic DNA probes identified 11 different genotypes of A. azollae isolates. These included three endosymbiotic genotypes within Azolla filiculoides species and two genotypes within both A. caroliniana and Azolla pinnata endosymbionts. Although we were not able to discriminate among anabaenas extracted from different ecotypes of Azolla nilotica, Azolla mexicina, Azolla rubra and Azolla microphylla species, each of the endosymbionts was easily identified as a unique genotype. When total DNA isolated from free-living Anabaena sp. strain PCC7120 was screened, none of the genomic DNA probes gave detectable positive hybridization. Total DNA of Nostoc cycas PCC7422 hybridized with six of eight genomic DNA fragments. These data imply that the dominant symbiotic organism in association with Azolla spp. is more closely related to Nostoc spp. than to free-living Anabaena spp. Images PMID:16348182

  8. 集胞藻PCC6803基因slr2049定点突变及其体内重组功能研究%Site-directed mutation of the gene slr2049 from Synechocy-stis sp.PCC 6803 and the study on the function

    Institute of Scientific and Technical Information of China (English)

    朱超; 陈思礼

    2013-01-01

    在Synechocystis sp.PCC 6803中找到与cpeS具有高度同源性的基因slr2049.采用PCR从Synechocystis sp.PCC 6803 DNA中扩增出cpcB,slr2049,hol,pcyA 4个基因;定点突变试剂盒构建8个突变体slr2049 (H21S)、slr2049 (L23S)、slr2049 (A24S)、slr2049 (F25S)、slr2049(W72L)、slr2049(G84S)、slr2049(R107S)和slr2049(Y124S);将它们与表达载体pCDFDuet-1和pET-23a(+)相连构建pCDF-cpcB-slr2049野生型和pCDF-cpcB-slr2049突变体以及pET-ho1-pcyA,进行高效表达,用亲和层析柱纯化,产物用SDS-PAGE和光谱检测.研究表明:突变体slr2049(H21S)、slr2049(L23S)、slr2049(F25S)、slr2049 (W72L)、slr2049 (G84S)、slr2049(Y124S)与野生型slr2049催化获得的重组藻蓝蛋白β亚基具有与天然藻蓝蛋白β亚基相似的光谱特性,其他2个突变体slr2049 (A24S)和slr2049(R107S)催化的产物则没有光谱特性.由此推测,第24位丙氨酸和第107位精氨酸可能是slr2049酶活性的必需氨基酸,其所处位置是slr2049的活性位点.%We have found gene slr2049 in Synechocystis sp.PCC 6803 which was homol ogous to the bilin lyase gene cpeS by some softwares for homologous analysis.Four genes were amplifed from DNA of Synechocystis sp.PCC 6803 by PCR.Eight mutants which were slr2049 (H21S),slr2049 (L23S),slr2049 (A24S),slr2049 (F25S),slr2049 (W72L),slr2049(G84S),slr2049(R107S)and sl-r2049(Y124S)were constructed by Mutan BEST Kit.pCDF-cpcB-slr2049,pCDF-cpcB-slr2049 mutants and pET-ho1-pcyA were formed from the linkage of four genes and mutants with pCDFDuet-land pET-23a (+).They were expressed and purified by affinity column.Proteins were detected by SDS-PAGE,absorption and fluorescence spectra.The results showed that phycocyanin β-subunit which had the same spectral characteristic with native phycocyanin β-subunit were catalysed by the protein encoded by gene slr2049 and mutants slr2049 (H21S),slr2049(L23S),slr2049(F25S),slr2049 (W72L),slr2049 (G84S),slr2049 (Y124S).The rest of mutants slr2049(A24S

  9. Open reading frame ssr2016 is required for antimycin A-sensitive photosystem I-driven cyclic electron flow in the cyanobacterium Synechocystis sp. PCC 6803

    NARCIS (Netherlands)

    Yeremenko, Nataliya; Jeanjean, Robert; Prommeenate, Peerada; Krasikov, Vladimir; Nixon, Peter J.; Vermaas, Wim F. J.; Havaux, Michel; Matthijs, Hans C. P.

    2005-01-01

    Open reading frame ssr2016 encodes a protein with substantial sequence similarities to PGR5 identified as a component of the antimycin A-sensitive ferredoxin:plastoquinone reductase (FQR) in PSI cyclic photophosphorylation in Arabidopsis thaliana. We studied cyclic electron flow in Synechocystis sp.

  10. The Nostoc punctiforme Genome

    Energy Technology Data Exchange (ETDEWEB)

    John C. Meeks

    2001-12-31

    Nostoc punctiforme is a filamentous cyanobacterium with extensive phenotypic characteristics and a relatively large genome, approaching 10 Mb. The phenotypic characteristics include a photoautotrophic, diazotrophic mode of growth, but N. punctiforme is also facultatively heterotrophic; its vegetative cells have multiple development alternatives, including terminal differentiation into nitrogen-fixing heterocysts and transient differentiation into spore-like akinetes or motile filaments called hormogonia; and N. punctiforme has broad symbiotic competence with fungi and terrestrial plants, including bryophytes, gymnosperms and an angiosperm. The shotgun-sequencing phase of the N. punctiforme strain ATCC 29133 genome has been completed by the Joint Genome Institute. Annotation of an 8.9 Mb database yielded 7432 open reading frames, 45% of which encode proteins with known or probable known function and 29% of which are unique to N. punctiforme. Comparative analysis of the sequence indicates a genome that is highly plastic and in a state of flux, with numerous insertion sequences and multilocus repeats, as well as genes encoding transposases and DNA modification enzymes. The sequence also reveals the presence of genes encoding putative proteins that collectively define almost all characteristics of cyanobacteria as a group. N. punctiforme has an extensive potential to sense and respond to environmental signals as reflected by the presence of more than 400 genes encoding sensor protein kinases, response regulators and other transcriptional factors. The signal transduction systems and any of the large number of unique genes may play essential roles in the cell differentiation and symbiotic interaction properties of N. punctiforme.

  11. Mutation of Gly195 of the ChlH subunit of Mg-chelatase reduces chlorophyll and further disrupts PS II assembly in a Ycf48-deficient strain of Synechocystis sp. PCC 6803

    Directory of Open Access Journals (Sweden)

    Tim Crawford

    2016-07-01

    Full Text Available Biogenesis of the photosystems in oxygenic phototrophs requires co-translational insertion of chlorophyll a. The first committed step of chlorophyll a biosynthesis is the insertion of a Mg2+ ion into the tetrapyrrole intermediate protoporphyrin IX, catalyzed by Mg-chelatase. We have identified a Synechocystis sp. PCC 6803 strain with a spontaneous mutation in chlH that results in a Gly195 to Glu substitution in a conserved region of the catalytic subunit of Mg-chelatase. Mutant strains containing the ChlH Gly195 to Glu mutation were generated using a two-step protocol that introduced the chlH gene into a putative neutral site in the chromosome prior to deletion of the native gene. The Gly195 to Glu mutation resulted in strains with decreased chlorophyll a. Deletion of the PS II assembly factor Ycf48 in a strain carrying the ChlH Gly195 to Glu mutation did not grow photoautotrophically. In addition, the ChlH-G195E:ΔYcf48 strain showed impaired PS II activity and decreased assembly of PS II centers in comparison to a ΔYcf48 strain. We suggest decreased chlorophyll in the ChlH-G195E mutant provides a background to screen for the role of assembly factors that are not essential under optimal growth conditions.

  12. Distinctive characters of Nostoc genomes in cyanolichens.

    Science.gov (United States)

    Gagunashvili, Andrey N; Andrésson, Ólafur S

    2018-06-05

    Cyanobacteria of the genus Nostoc are capable of forming symbioses with a wide range of organism, including a diverse assemblage of cyanolichens. Only certain lineages of Nostoc appear to be able to form a close, stable symbiosis, raising the question whether symbiotic competence is determined by specific sets of genes and functionalities. We present the complete genome sequencing, annotation and analysis of two lichen Nostoc strains. Comparison with other Nostoc genomes allowed identification of genes potentially involved in symbioses with a broad range of partners including lichen mycobionts. The presence of additional genes necessary for symbiotic competence is likely reflected in larger genome sizes of symbiotic Nostoc strains. Some of the identified genes are presumably involved in the initial recognition and establishment of the symbiotic association, while others may confer advantage to cyanobionts during cohabitation with a mycobiont in the lichen symbiosis. Our study presents the first genome sequencing and genome-scale analysis of lichen-associated Nostoc strains. These data provide insight into the molecular nature of the cyanolichen symbiosis and pinpoint candidate genes for further studies aimed at deciphering the genetic mechanisms behind the symbiotic competence of Nostoc. Since many phylogenetic studies have shown that Nostoc is a polyphyletic group that includes several lineages, this work also provides an improved molecular basis for demarcation of a Nostoc clade with symbiotic competence.

  13. The antisense RNA As1_flv4 in the Cyanobacterium Synechocystis sp. PCC 6803 prevents premature expression of the flv4-2 operon upon shift in inorganic carbon supply.

    Science.gov (United States)

    Eisenhut, Marion; Georg, Jens; Klähn, Stephan; Sakurai, Isamu; Mustila, Henna; Zhang, Pengpeng; Hess, Wolfgang R; Aro, Eva-Mari

    2012-09-28

    The functional relevance of natural cis-antisense transcripts is mostly unknown. Here we have characterized the association of three antisense RNAs and one intergenically encoded noncoding RNA with an operon that plays a crucial role in photoprotection of photosystem II under low carbon conditions in the cyanobacterium Synechocystis sp. PCC 6803. Cyanobacteria show strong gene expression dynamics in response to a shift of cells from high carbon to low levels of inorganic carbon (C(i)), but the regulatory mechanisms are poorly understood. Among the most up-regulated genes in Synechocystis are flv4, sll0218, and flv2, which are organized in the flv4-2 operon. The flavodiiron proteins encoded by this operon open up an alternative electron transfer route, likely starting from the Q(B) site in photosystem II, under photooxidative stress conditions. Our expression analysis of cells shifted from high carbon to low carbon demonstrated an inversely correlated transcript accumulation of the flv4-2 operon mRNA and one antisense RNA to flv4, designated as As1_flv4. Overexpression of As1_flv4 led to a decrease in flv4-2 mRNA. The promoter activity of as1_flv4 was transiently stimulated by C(i) limitation and negatively regulated by the AbrB-like transcription regulator Sll0822, whereas the flv4-2 operon was positively regulated by the transcription factor NdhR. The results indicate that the tightly regulated antisense RNA As1_flv4 establishes a transient threshold for flv4-2 expression in the early phase after a change in C(i) conditions. Thus, it prevents unfavorable synthesis of the proteins from the flv4-2 operon.

  14. The Antisense RNA As1_flv4 in the Cyanobacterium Synechocystis sp. PCC 6803 Prevents Premature Expression of the flv4-2 Operon upon Shift in Inorganic Carbon Supply*

    Science.gov (United States)

    Eisenhut, Marion; Georg, Jens; Klähn, Stephan; Sakurai, Isamu; Mustila, Henna; Zhang, Pengpeng; Hess, Wolfgang R.; Aro, Eva-Mari

    2012-01-01

    The functional relevance of natural cis-antisense transcripts is mostly unknown. Here we have characterized the association of three antisense RNAs and one intergenically encoded noncoding RNA with an operon that plays a crucial role in photoprotection of photosystem II under low carbon conditions in the cyanobacterium Synechocystis sp. PCC 6803. Cyanobacteria show strong gene expression dynamics in response to a shift of cells from high carbon to low levels of inorganic carbon (Ci), but the regulatory mechanisms are poorly understood. Among the most up-regulated genes in Synechocystis are flv4, sll0218, and flv2, which are organized in the flv4-2 operon. The flavodiiron proteins encoded by this operon open up an alternative electron transfer route, likely starting from the QB site in photosystem II, under photooxidative stress conditions. Our expression analysis of cells shifted from high carbon to low carbon demonstrated an inversely correlated transcript accumulation of the flv4-2 operon mRNA and one antisense RNA to flv4, designated as As1_flv4. Overexpression of As1_flv4 led to a decrease in flv4-2 mRNA. The promoter activity of as1_flv4 was transiently stimulated by Ci limitation and negatively regulated by the AbrB-like transcription regulator Sll0822, whereas the flv4-2 operon was positively regulated by the transcription factor NdhR. The results indicate that the tightly regulated antisense RNA As1_flv4 establishes a transient threshold for flv4-2 expression in the early phase after a change in Ci conditions. Thus, it prevents unfavorable synthesis of the proteins from the flv4-2 operon. PMID:22854963

  15. ORF Alignment: NC_003272 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available bosomal protein L22 ... [Nostoc sp. PCC 7120] ... Length = 111 ... Query: 7 ... EVKAIARFIRMS...PYKVRRVLDQIRGLSYREALIILEFMPYRATEPVLTLLRSAAANAEHN 66 ... EVKAIARFIRMSPYKVRRVLDQIRG...LSYREALIILEFMPYRATEPVLTLLRSAAANAEHN Sbjct: 1 ... EVKAIARFIRMSPYKVRRVLDQIRGLSYREALIILEFMPYRATEPVLTLLRSAAANAEHN 60 ...

  16. ORF Alignment: NC_003272 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available stidine kinase ... [Nostoc sp. PCC 7120] ... Length = 73 ... Query: 410 ADLIQLNRLKDEFLACISH...ELKTPLTAVLGLSRLLVDQQLGELNERQARYAGLIHQSGRH 469 ... ADLIQLNRLKDEFLACISHELKTPLTAVLG...LSRLLVDQQLGELNERQARYAGLIHQSGRH Sbjct: 1 ... ADLIQLNRLKDEFLACISHELKTPLTAVLGLSRLLVDQQLGELNERQARYAGLIHQSGRH 60 ...

  17. ORF Alignment: NC_003272 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ... family [Nostoc sp. PCC 7120] ... Length = 102 ... Query: 48 ... IGIDLEYLRPTSD...LESLAKRFFLPREYELLRSLPDEQKQKIFFRYWTCKEAYLKATGDGI 107 ... IGIDLEYLRPTSDLESLAKRFFLPREYELLRSLPDEQ...KQKIFFRYWTCKEAYLKATGDGI Sbjct: 1 ... IGIDLEYLRPTSDLESLAKRFFLPREYELLRSLPDEQKQKIFFRYWTCKEAYLKATGDGI 60 ...

  18. ORF Alignment: NC_003272 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ... - Nostoc sp. (strain PCC 7120) ... Length = 66 ... Query: 5 ... ESQTPVTLSDRELQIIDLVAAGLTNQEIAAKLEISKRTVDNHIS...NILDKTRTENRVALVR 64 ... ESQTPVTLSDRELQIIDLVAAGLTNQEIAAKLEISKRTVDNHISNILDKTRTENRVALVR Sbjct: 1 ... ESQTPVTLSDRELQIIDLVAAGLTNQEIAAKLEISKRTVDNHISNILDKTRTENRVALVR 60 ...

  19. Induction of the Nitrate Assimilation nirA Operon and Protein-Protein Interactions in the Maturation of Nitrate and Nitrite Reductases in the Cyanobacterium Anabaena sp. Strain PCC 7120.

    Science.gov (United States)

    Frías, José E; Flores, Enrique

    2015-07-01

    Nitrate is widely used as a nitrogen source by cyanobacteria, in which the nitrate assimilation structural genes frequently constitute the so-called nirA operon. This operon contains the genes encoding nitrite reductase (nirA), a nitrate/nitrite transporter (frequently an ABC-type transporter; nrtABCD), and nitrate reductase (narB). In the model filamentous cyanobacterium Anabaena sp. strain PCC 7120, which can fix N2 in specialized cells termed heterocysts, the nirA operon is expressed at high levels only in media containing nitrate or nitrite and lacking ammonium, a preferred nitrogen source. Here we examined the genes downstream of the nirA operon in Anabaena and found that a small open reading frame of unknown function, alr0613, can be cotranscribed with the operon. The next gene in the genome, alr0614 (narM), showed an expression pattern similar to that of the nirA operon, implying correlated expression of narM and the operon. A mutant of narM with an insertion mutation failed to produce nitrate reductase activity, consistent with the idea that NarM is required for the maturation of NarB. Both narM and narB mutants were impaired in the nitrate-dependent induction of the nirA operon, suggesting that nitrite is an inducer of the operon in Anabaena. It has previously been shown that the nitrite reductase protein NirA requires NirB, a protein likely involved in protein-protein interactions, to attain maximum activity. Bacterial two-hybrid analysis confirmed possible NirA-NirB and NarB-NarM interactions, suggesting that the development of both nitrite reductase and nitrate reductase activities in cyanobacteria involves physical interaction of the corresponding enzymes with their cognate partners, NirB and NarM, respectively. Nitrate is an important source of nitrogen for many microorganisms that is utilized through the nitrate assimilation system, which includes nitrate/nitrite membrane transporters and the nitrate and nitrite reductases. Many cyanobacteria

  20. A hydrogen-producing, hydrogenase-free mutant strain of Nostoc punctiforme ATCC 29133

    Energy Technology Data Exchange (ETDEWEB)

    Lindberg, P.; Lindblad, P. [Uppsala Univ. (Sweden). Dept. of Physiological Botany; Schuetz, K.; Happe, T. [Universitaet Bonn (Germany). Botanisches Inst.

    2002-12-01

    The hupL gene, encoding the uptake hydrogenase large subunit, in Nostoc sp. strain ATCC 29133, a strain lacking a bidirectional hydrogenase, was inactivated by insertional mutagenesis. Recombinant strains were isolated and analysed, and one hupL{sup -} strain, NHM5, was selected for further study. Cultures of NHM5 were grown under nitrogen-fixing conditions and H{sub 2} evolution under air was observed using an H{sub 2} electrode. (Author)

  1. Identification and topographical characterisation of microbial nanowires in Nostoc punctiforme.

    Science.gov (United States)

    Sure, Sandeep; Torriero, Angel A J; Gaur, Aditya; Li, Lu Hua; Chen, Ying; Tripathi, Chandrakant; Adholeya, Alok; Ackland, M Leigh; Kochar, Mandira

    2016-03-01

    Extracellular pili-like structures (PLS) produced by cyanobacteria have been poorly explored. We have done detailed topographical and electrical characterisation of PLS in Nostoc punctiforme PCC 73120 using transmission electron microscopy (TEM) and conductive atomic force microscopy (CAFM). TEM analysis showed that N. punctiforme produces two separate types of PLS differing in their length and diameter. The first type of PLS are 6-7.5 nm in diameter and 0.5-2 µm in length (short/thin PLS) while the second type of PLS are ~20-40 nm in diameter and more than 10 µm long (long/thick PLS). This is the first study to report long/thick PLS in N. punctiforme. Electrical characterisation of these two different PLS by CAFM showed that both are electrically conductive and can act as microbial nanowires. This is the first report to show two distinct PLS and also identifies microbial nanowires in N. punctiforme. This study paves the way for more detailed investigation of N. punctiforme nanowires and their potential role in cell physiology and symbiosis with plants.

  2. Enhancement of soybean (Glycine max L.) growth by bio-fertilizers of Nostoc muscorum and Nostoc rivulare

    International Nuclear Information System (INIS)

    Sholkamy, E.N.; Komy, H.M.E.

    2015-01-01

    In the present study the nitrogenase activity of Nostoc muscorum and Nostoc rivulare was evaluated in vitro; the test showed that Nostoc muscorum and Nostoc rivulare have the ability to fix nitrogen. In a pot experiment under field conditions, the results of the present study showed that inoculation of the soybean plant with Nostoc muscorum and Nostoc rivulare, either alone or in combination with N-fertilizer at 50 and 100 kg N/ha, caused a significant increase in the growth of these plants, as reflected in plant height, leaf area, weight of plant as well as the legume weight of soybeans. The combination of biofertilization and N-fertilization, especially at 100 kg N/ha, had more effect on both the growth of soybeans and nitrogenase activity compared to biofertilization alone. Nostoc muscorum and Nostoc rivulare are a promising biofertilizers for achieving an efficient association between N2 fixing cyanobacteria and soybeans; and thus enhancement of the growth. (author)

  3. Consumption of fa cai Nostoc soup: a potential for BMAA exposure from Nostoc cyanobacteria in China?

    Science.gov (United States)

    Roney, Britton R; Renhui, Li; Banack, Sandra Anne; Murch, Susan; Honegger, Rosmarie; Cox, Paul Alan

    2009-01-01

    Grown in arid regions of western China the cyanobacterium Nostoc flagelliforme--called fa cai in Mandarin and fat choy in Cantonese--is wild-harvested and used to make soup consumed during New Year's celebrations. High prices, up to $125 USD/kg, led to overharvesting in Inner Mongolia, Ningxia, Gansu, Qinghai, and Xinjiang. Degradation of arid ecosystems, desertification, and conflicts between Nostoc harvesters and Mongol herdsmen concerned the Chinese environmental authorities, leading to a government ban of Nostoc commerce. This ban stimulated increased marketing of a substitute made from starch. We analysed samples purchased throughout China as well as in Chinese markets in the United States and the United Kingdom. Some were counterfeits consisting of dyed starch noodles. A few samples from California contained Nostoc flagelliforme but were adulterated with starch noodles. Other samples, including those from the United Kingdom, consisted of pure Nostoc flagelliforme. A recent survey of markets in Cheng Du showed no real Nostoc flagelliforme to be marketed. Real and artificial fa cai differ in the presence of beta-N-methylamino-L-alanine (BMAA). Given its status as a high-priced luxury food, the government ban on collection and marketing, and the replacement of real fa cai with starch substitutes consumed only on special occasions, it is anticipated that dietary exposure to BMAA from fa cai will be reduced in the future in China.

  4. Cell wall amidase AmiC1 is required for cellular communication and heterocyst development in the cyanobacterium Anabaena PCC 7120 but not for filament integrity.

    Science.gov (United States)

    Berendt, Susanne; Lehner, Josef; Zhang, Yao Vincent; Rasse, Tobias M; Forchhammer, Karl; Maldener, Iris

    2012-10-01

    Filamentous cyanobacteria of the order Nostocales display typical properties of multicellular organisms. In response to nitrogen starvation, some vegetative cells differentiate into heterocysts, where fixation of N(2) takes place. Heterocysts provide a micro-oxic compartment to protect nitrogenase from the oxygen produced by the vegetative cells. Differentiation involves fundamental remodeling of the gram-negative cell wall by deposition of a thick envelope and by formation of a neck-like structure at the contact site to the vegetative cells. Cell wall-hydrolyzing enzymes, like cell wall amidases, are involved in peptidoglycan maturation and turnover in unicellular bacteria. Recently, we showed that mutation of the amidase homologue amiC2 gene in Nostoc punctiforme ATCC 29133 distorts filament morphology and function. Here, we present the functional characterization of two amiC paralogues from Anabaena sp. strain PCC 7120. The amiC1 (alr0092) mutant was not able to differentiate heterocysts or to grow diazotrophically, whereas the amiC2 (alr0093) mutant did not show an altered phenotype under standard growth conditions. In agreement, fluorescence recovery after photobleaching (FRAP) studies showed a lack of cell-cell communication only in the AmiC1 mutant. Green fluorescent protein (GFP)-tagged AmiC1 was able to complement the mutant phenotype to wild-type properties. The protein localized in the septal regions of newly dividing cells and at the neck region of differentiating heterocysts. Upon nitrogen step-down, no mature heterocysts were developed in spite of ongoing heterocyst-specific gene expression. These results show the dependence of heterocyst development on amidase function and highlight a pivotal but so far underestimated cellular process, the remodeling of peptidoglycan, for the biology of filamentous cyanobacteria.

  5. Lietuvos Nostoc ir Desmonostoc genčių melsvabakterių morfologija, ekologija ir filogenija

    OpenAIRE

    Špakaitė, Ina

    2014-01-01

    The aim of the study was to investigate the morphology, ecology and phylogeny of cyanobacteria belonging to genera Nostoc and Desmonostoc in Lithuania. The detailed research of freshwater and terrestrial Nostoc and Desmonostoc species provided new data on taxonomy, biology and ecology of these cyanobacteria and the overall diversity of algae in Lithuania. 20 Nostoc species and two intraspecific taxa, and 18 taxa to the Nostoc genus level were identified. Twelve Nostoc species and intraspecifi...

  6. Morphology, ecology and phylogeny of cyanobacteria belonging to genera Nostoc and Desmonostoc in Lithuania

    OpenAIRE

    Špakaitė, Ina

    2014-01-01

    The aim of the study was to investigate the morphology, ecology and phylogeny of cyanobacteria belonging to genera Nostoc and Desmonostoc in Lithuania. The detailed research of freshwater and terrestrial Nostoc and Desmonostoc species provided new data on taxonomy, biology and ecology of these cyanobacteria and the overall diversity of algae in Lithuania. 20 Nostoc species and two intraspecific taxa, and 18 taxa to the Nostoc genus level were identified. Twelve Nostoc species and intraspecifi...

  7. comparative transcriptomics between Synechococcus PCC 7942 and Synechocystis PCC 6803 provide insights into mechanisms of adaptation to stress.

    Energy Technology Data Exchange (ETDEWEB)

    Konstantinos, Billis [USDOE Joint Genome Institute (JGI), Walnut Creek, CA (United States); European Bioinformatics Inst., Hinxton, Cambridge (United Kingdom). European Molecular Biology Lab.; Aristotle Univ., Thessaloniki (Greece). Dept. of Genetics; Billini, Maria [USDOE Joint Genome Institute (JGI), Walnut Creek, CA (United States); Max Planck Inst. for Terrestrial Microbiology, Marburg (Germany); Tripp, Harry J. [USDOE Joint Genome Institute (JGI), Walnut Creek, CA (United States); Kyrpides, Nikos C. [USDOE Joint Genome Institute (JGI), Walnut Creek, CA (United States); Mavrommatis, Konstantinos [USDOE Joint Genome Institute (JGI), Walnut Creek, CA (United States); Celgene Corp, San Francisco, CA (United States)

    2014-03-21

    Background: Synechococcus sp. PCC 7942 and Synechocystis sp. PCC 6803 are model cyanobacteria from which the metabolism and adaptive responses of other cyanobacteria are inferred. Here we report the gene expression response of these two strains to a variety of nutrient and environmental stresses of varying duration, using transcriptomics. Our data comprise both stranded and 5? enriched libraries in order to elucidate many aspects of the transcriptome. Results: Both organisms were exposed to stress conditions due to nutrient deficiency (inorganic carbon) or change of environmental conditions (salinity, temperature, pH, light) sampled at 1 and 24 hours after the application of stress. The transcriptome profile of each strain revealed similarities and differences in gene expression for photosynthetic and respiratory electron transport chains and carbon fixation. Transcriptome profiles also helped us improve the structural annotation of the genome and identify possible missed genes (including anti-sense) and determine transcriptional units (operons). Finally, we predicted association of proteins of unknown function biochemical pathways by associating them to well-characterized ones based on their transcript levels correlation. Conclusions: Overall, this study results an informative annotation of those species and the comparative analysis of the response of the two organisms revealed similarities but also significant changes in the way they respond to external stress and the duration of the response

  8. Penggunaan precipitated calcium carbonate (PCC sebagai filler untuk sol karet sepatu olah raga

    Directory of Open Access Journals (Sweden)

    Herminiwati

    2010-12-01

    Full Text Available Abstract The objective of the research was to investigate the utilization of Precipitated Calcium Carbonate (PCC as filler in producing sport shoe rubber soles. PCC is a white filler needed for production of nonblack colour rubber products. There are four types of PCC that have been used including two local PCC from Wonosari and East Java, and two imported PCC from Japan and Taiwan. The amount of PCC added into the sport shoe sole rubber compound was varied in 30,45,60,75 and 90 per hundred rubber (phr. The compounding was carried-out by using two roll mills machine, and the compound was subsequently measured their optimum vulcanization time by using rheometer. The produced compound was then subjected to vulcanistion process by using hydrolic press at temperature 1500C and pressure 150 kg/ cm2. The quality of shoes sole vulcanisates were compare to standard quality of SNI. 12-7075-2005 about cemented system sport shoes. The results indicated that the best formula of rubber compound for sport shoes sole were made by using NR 80 phr, NBR 20 phr, paraffinic oil 10 phr, aluminium silicate 30 phr, ZnO 5 phr, TiO2 10 phr, stearic acid 1 phr, vulkanox SP 1 phr, paraffin wax 1 phr, TMTD 0,5 phr, CBS 2 phr, sulphur 1,2 phr with the amount of PCC Actifort 700 of 45 phr. The best formula meet the requirement SNI 12-7075-2005 and they were characterized by tensile sterength 16,79 N/mm2, elongation at break 529,92% tear resistance 9,06 N/mm2, specific gravity 1,28 g/cm3, hardness 55 shore A, Grasselli absrassion resistancing filler. The local PCC from Wonosari can be used for substitution of the imported PCC as the white filler for the production of rubber compound sport shoes sole. However, particle size reduction and coating or surface treatment of local PCC were needed for improving the quality and the role of reinforcing filler.

  9. Protein (Cyanobacteria): 285307 [PGDBj - Ortholog DB

    Lifescience Database Archive (English)

    Full Text Available ith PAS/PAC and Chase2 sensors Nostoc sp. PCC 7107 MSKQLGKSFVSSNLNLNLKQLLDRKYRQLVVAFSVAVCIILLRSVGMFQSLELAGLD...YP_007048593.1 1117:4890 1161:684 1162:948 1177:381 317936:58 diguanylate cyclase w

  10. Protein (Cyanobacteria): 504928995 [PGDBj - Ortholog DB

    Lifescience Database Archive (English)

    Full Text Available WP_015116097.1 ... 1117:4682 ... 1161:4218 ... 1162:3122 1177:1104 317936:3404 ... NADPH-d...ependent 7-cyano-7-deazaguanine reductase Nostoc sp. PCC 7107 MTQDTSEVKYGERNIAEGNLITFPNPRVGRRYDINITLPEFTCKCP

  11. Metabolism and resources of spherical colonies of Nostoc zetterstedtii

    DEFF Research Database (Denmark)

    Jensen, Kaj Sand; Raun, Ane-Marie Løvendahl; Borum, Jens

    2009-01-01

    Constraints imposed by the spherical form and gelatinous matrix of centimeter-thick colonies of the cyanobacterium Nostoc zetterstedtii on its functional properties were tested by examining the scaling of its composition, light absorption, photosynthesis, and respiration to individual size....... Nostoc absorbed 96% of incident light from the surface to the center because of high areal pigment density, but absorbed photons were used with low quantum efficiency (11- 38 mmol O2 mol21 photon) and photosynthesis was low relative to dark respiration (2.0-5.4). Therefore, N. zetterstedtii is threatened...... by reduced light availability and only extended to lake depths receiving about 12% of surface irradiance, whereas mosses, characeans, and angiosperms with thin photosynthetic tissues grew deeper (3.1-7.5% of surface irradiance). Nostoc ameliorated the restrictions of low lake DIC and long diffusion paths...

  12. Growth response to temperature and irradiance in Nostoc spongiaeforme

    Science.gov (United States)

    California water-seeded rice fields are typically shallow and have high nutrient levels, which are ideal growing conditions for algae and cyanobacteria. Nostoc spongiaeforme is problematic in California rice fields because floating mats may dislodge seedlings or smother them when the mats accumulat...

  13. Understanding nutrient exchange between Azolla and its symbiont, Nostoc

    OpenAIRE

    Eily, Ariana

    2017-01-01

    This is an in-depth look at the research I am doing for my doctoral degree at Duke University, investigating the exchange of nutrients between the aquatic fern genus, Azolla, and its cyanobacterial symbiont, Nostoc azollae. All of the illustrations and microscopy images within this presentation are my own.

  14. Comparison of plasmids from the cyanobacterium Nostoc PCC 7524 with two mutant strains unable to form heterocysts

    NARCIS (Netherlands)

    Reaston, J.; Hondel, C.A.M.J.J. van den; Ende, A. van der; Arkel, G.A. van; Stewart, W.D.P.; Herdman, M.

    1980-01-01

    Cyanobacteria (bluegreen bacteria) are O₂-evolving photosynthetic prokaryotes some species of which fix N₂ in air because the nitrogenase is protected from O₂ inactivation by being localized in differentiated cells called heterocysts. Recently much attention has been paid to the possible role

  15. PCC/SRC, PCC and SRC Calculation from Multivariate Input for Sensitivity Analysis

    International Nuclear Information System (INIS)

    Iman, R.L.; Shortencarier, M.J.; Johnson, J.D.

    1995-01-01

    1 - Description of program or function: PCC/SRC is designed for use in conjunction with sensitivity analyses of complex computer models. PCC/SRC calculates the partial correlation coefficients (PCC) and the standardized regression coefficients (SRC) from the multivariate input to, and output from, a computer model. 2 - Method of solution: PCC/SRC calculates the coefficients on either the original observations or on the ranks of the original observations. These coefficients provide alternative measures of the relative contribution (importance) of each of the various input variables to the observed variations in output. Relationships between the coefficients and differences in their interpretations are identified. If the computer model output has an associated time or spatial history, PCC/SRC will generate a graph of the coefficients over time or space for each input-variable, output- variable combination of interest, indicating the importance of each input value over time or space. 3 - Restrictions on the complexity of the problem - Maxima of: 100 observations, 100 different time steps or intervals between successive dependent variable readings, 50 independent variables (model input), 20 dependent variables (model output). 10 ordered triples specifying intervals between dependent variable readings

  16. Antagonistic interactions between filamentous heterotrophs and the cyanobacterium Nostoc muscorum

    Directory of Open Access Journals (Sweden)

    Wolf Sarah

    2011-09-01

    Full Text Available Abstract Background Little is known about interactions between filamentous heterotrophs and filamentous cyanobacteria. Here, interactions between the filamentous heterotrophic bacteria Fibrella aestuarina (strain BUZ 2 and Fibrisoma limi (BUZ 3 with an axenic strain of the autotrophic filamentous cyanobacterium Nostoc muscorum (SAG 25.82 were studied in mixed cultures under nutrient rich (carbon source present in medium and poor (carbon source absent in medium conditions. Findings F. aestuarina BUZ 2 significantly reduced the cyanobacterial population whereas F. limi BUZ 3 did not. Physical contact between heterotrophs and autotroph was observed and the cyanobacterial cells showed some level of damage and lysis. Therefore, either contact lysis or entrapment with production of extracellular compounds in close vicinity of host cells could be considered as potential modes of action. The supernatants from pure heterotrophic cultures did not have an effect on Nostoc cultures. However, supernatant from mixed cultures of BUZ 2 and Nostoc had a negative effect on cyanobacterial growth, indicating that the lytic compounds were only produced in the presence of Nostoc. The growth and survival of tested heterotrophs was enhanced by the presence of Nostoc or its metabolites, suggesting that the heterotrophs could utilize the autotrophs and its products as a nutrient source. However, the autotroph could withstand and out-compete the heterotrophs under nutrient poor conditions. Conclusions Our results suggest that the nutrients in cultivation media, which boost or reduce the number of heterotrophs, were the important factor influencing the outcome of the interplay between filamentous heterotrophs and autotrophs. For better understanding of these interactions, additional research is needed. In particular, it is necessary to elucidate the mode of action for lysis by heterotrophs, and the possible defense mechanisms of the autotrophs.

  17. Genetic Diversity of Nostoc Symbionts Endophytically Associated with Two Bryophyte Species

    OpenAIRE

    Costa, José-Luis; Paulsrud, Per; Rikkinen, Jouko; Lindblad, Peter

    2001-01-01

    The diversity of the endophytic Nostoc symbionts of two thalloid bryophytes, the hornwort Anthoceros fusiformis and the liverwort Blasia pusilla, was examined using the tRNALeu (UAA) intron sequence as a marker. The results confirmed that many different Nostoc strains are involved in both associations under natural conditions in the field. The level of Nostoc diversity within individual bryophyte thalli varied, but single DNA fragments were consistently amplified from individual symbiotic col...

  18. Control of light-dependent keto carotenoid biosynthesis in Nostoc 7120 by the transcription factor NtcA.

    Science.gov (United States)

    Sandmann, Gerhard; Mautz, Jürgen; Breitenbach, Jürgen

    2016-09-01

    In Nostoc PCC 7120, two different ketolases, CrtW and CrtO are involved in the formation of keto carotenoids from β-carotene. In contrast to other cyanobacteria, CrtW catalyzes the formation of monoketo echinenone whereas CrtO is the only enzyme for the synthesis of diketo canthaxanthin. This is the major photo protective carotenoid in this cyanobacterium. Under high-light conditions, basic canthaxanthin formation was transcriptionally up-regulated. Upon transfer to high light, the transcript levels of all investigated carotenogenic genes including those coding for phytoene synthase, phytoene desaturase and both ketolases were increased. These transcription changes proceeded via binding of the transcription factor NtcA to the promoter regions of the carotenogenic genes. The binding was absolutely dependent on the presence of reductants and oxo-glutarate. Light-stimulated transcript formation was inhibited by DCMU. Therefore, photosynthetic electron transport is proposed as the sensor for high-light and a changing redox state as a signal for NtcA binding.

  19. Characterization of a novel debranching enzyme from Nostoc punctiforme possessing a high specificity for long branched chains

    International Nuclear Information System (INIS)

    Choi, Ji-Hye; Lee, Heeseob; Kim, Young-Wan; Park, Jong-Tae; Woo, Eui-Jeon; Kim, Myo-Jeong; Lee, Byong-Hoon; Park, Kwan-Hwa

    2009-01-01

    A novel debranching enzyme from Nostoc punctiforme PCC73102 (NPDE) exhibits hydrolysis activity toward both α-(1,6)- and α-(1,4)-glucosidic linkages. The action patterns of NPDE revealed that branched chains are released first, and the resulting maltooligosaccharides are then hydrolyzed. Analysis of the reaction with maltooligosaccharide substrates labeled with 14 C-glucose at the reducing end shows that NPDE specifically liberates glucose from the reducing end. Kinetic analyses showed that the hydrolytic activity of NPDE is greatly affected by the length of the substrate. The catalytic efficiency of NPDE increased considerably upon using substrates that can occupy at least eight glycone subsites such as maltononaose and maltooctaosyl-α-(1,6)-β-cyclodextrin. These results imply that NPDE has a unique subsite structure consisting of -8 to +1 subsites. Given its unique subsite structure, side chains shorter than maltooctaose in amylopectin were resistant to hydrolysis by NPDE, and the population of longer side chains was reduced.

  20. Light influences cytokinin biosynthesis and sensing in Nostoc (cyanobacteria)

    Czech Academy of Sciences Publication Activity Database

    Frébortová, J.; Plíhal, O.; Florová, P.; Kokáš, F.; Kubiasová, K.; Greplová, M.; Šimura, J.; Novák, Ondřej; Frébort, I.

    2017-01-01

    Roč. 53, č. 3 (2017), s. 703-714 ISSN 0022-3646 R&D Projects: GA MŠk(CZ) LO1204 Institutional support: RVO:61389030 Keywords : cytokinin * expression profiling * miaA * Nostoc * RNA-seq * sensor kinase * tRNA IPT Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Plant sciences, botany Impact factor: 2.608, year: 2016

  1. Studies of polysaccharides from three edible species of Nostoc (cyanobacteria) with different colony morphologies : structural characterization and effect on the complement system of polysaccharides from Nostoc commune

    NARCIS (Netherlands)

    Brüll, L.P.; Huang, Z.; Thomas-Oates, J.E.; Smestad-Paulsen, B.; Cohen, E.H.; Michaelsen, T.E.

    2000-01-01

    The cyanobacterium Nostoc commune Vaucher produces quite complex extracellular polysaccharides. The cyanobacterium is nitrogen fixing, and on growing the cyanobacterium in media with and without nitrogen, different types of extracellular polysaccharides were obtained. These were also different from

  2. Photo- and heterotrophic nitrogenase activity by the cyano-bacterium Nostoc in symbiosis with the bryophyte Anthoceros

    International Nuclear Information System (INIS)

    Steinberg, N.A.; Meeks, J.C.

    1987-01-01

    In symbiosis with Anthoceros, Nostoc is thought to do little or no photosynthesis. However, light-dependent 14 CO 2 fixation by symbiotic Nostoc, freshly isolated from pure cultures of the reconstituted Anthoceros-Nostoc association, was 16% of that by free-living Nostoc. A DCMU-resistant mutant of Nostoc was isolated that fixed CO 2 at rates comparable to wild-type in both symbiotic and free-living growth states. To determine if symbiotic Nostoc can use its photosynthate directly to fix nitrogen, acetylene reduction by Anthoceros associations reconstituted with wild-type Nostoc was compared to associations with the DCMU-resistant mutant. In wild-type Anthoceros-Nostoc acetylene reduction was inhibited 97% by 5 μM DCMU, while inhibition of the DCMU-resistant Nostoc association was only 63%. Additions of glucose, fructose, maltose or sucrose to wild-type associations completely restored DCMU-inhibited acetylene reduction in the light. Acetylene reduction in the dark was stimulated by glucose, attaining 84% of the uninhibited light-dependent value. The authors conclude that symbiotic Nostoc maintains a pool of photosynthate which supports nitrogenase activity. The pool can also be supplemented from plant sources

  3. Protein (Cyanobacteria): 35831 [PGDBj - Ortholog DB

    Lifescience Database Archive (English)

    Full Text Available etical protein Npun_F5100 Nostoc punctiforme PCC 73102 MKTLVNLTQQSVVGEIESVLDTYPYHPYQKAFAIPDLRQELIVFVLTRLPSFDGAMSEGHISLAEAEQGSLAYYKLPRKPLEQQLHLQNLIHQGICLIVQEKSDWINDRVCEIVQPACEASHWFG ...

  4. Genomic Changes Associated with the Evolutionary Transitions of Nostoc to a Plant Symbiont

    Science.gov (United States)

    Liaimer, Anton; Pederson, Eric; Kim, Sea-Yong; Shapiro, Nicole; Woyke, Tanja; Altermark, Bjørn; Pawlowski, Katharina; Weyman, Philip D; Dupont, Christopher L

    2018-01-01

    Abstract Cyanobacteria belonging to the genus Nostoc comprise free-living strains and also facultative plant symbionts. Symbiotic strains can enter into symbiosis with taxonomically diverse range of host plants. Little is known about genomic changes associated with evolutionary transition of Nostoc from free-living to plant symbiont. Here, we compared the genomes derived from 11 symbiotic Nostoc strains isolated from different host plants and infer phylogenetic relationships between strains. Phylogenetic reconstructions of 89 Nostocales showed that symbiotic Nostoc strains with a broad host range, entering epiphytic and intracellular or extracellular endophytic interactions, form a monophyletic clade indicating a common evolutionary history. A polyphyletic origin was found for Nostoc strains which enter only extracellular symbioses, and inference of transfer events implied that this trait was likely acquired several times in the evolution of the Nostocales. Symbiotic Nostoc strains showed enriched functions in transport and metabolism of organic sulfur, chemotaxis and motility, as well as the uptake of phosphate, branched-chain amino acids, and ammonium. The genomes of the intracellular clade differ from that of other Nostoc strains, with a gain/enrichment of genes encoding proteins to generate l-methionine from sulfite and pathways for the degradation of the plant metabolites vanillin and vanillate, and of the macromolecule xylan present in plant cell walls. These compounds could function as C-sources for members of the intracellular clade. Molecular clock analysis indicated that the intracellular clade emerged ca. 600 Ma, suggesting that intracellular Nostoc symbioses predate the origin of land plants and the emergence of their extant hosts. PMID:29554291

  5. A new UV-A/B protecting pigment in the terrestrial cyanobacterium Nostoc commune

    International Nuclear Information System (INIS)

    Scherer, S.; Chen, T.W.; Boeger, P.

    1988-01-01

    A new ultraviolet (UV)-A/B absorbing pigment with maxima at 312 and 330 nanometers from the cosmopolitan terrestrial cyanobacterium Nostoc commune is described. The pigment is found in high amounts (up to 10% of dry weight) in colonies grown under solar UV radiation but only in low concentrations in laboratory cultures illuminated by artificial light without UV. Its experimental induction by UV as well as its capacity to efficiently protect Nostoc against UV radiation is reported

  6. Genetic diversity of Nostoc microsymbionts from Gunnera tinctoria revealed by PCR-STRR fingerprinting.

    Science.gov (United States)

    Guevara, R; Armesto, J J; Caru, M

    2002-08-01

    The cyanobacteria belonging to the genus Nostoc fix atmospheric nitrogen, both as free-living organisms and in symbiotic associations with a wide range of hosts, including bryophytes, gymnosperms (cycads), the small water fern Azolla (Pteridophyte), the angiosperm genus Gunnera, and fungi (lichens). The Gunnera-Nostoc symbiosis is the only one that involves a flowering plant. In Chile, 12 species of Gunnera have been described with a broad distribution in the temperate region. We examined the genetic diversity of Nostoc symbionts from three populations of Gunnera tinctoria from Abtao, Chiloé Island, southern Chile, and microsymbionts from other two species of Gunnera from southern Chile, using PCR amplification of STRR (short tandemly repeated repetitive) sequences of the Nostoc infected tissue. To our knowledge, this is the first report of PCR fingerprinting obtained directly from symbiotic tissue of Gunnera. Genetic analyses revealed that Nostoc symbionts exhibit important genetic diversity among host plants, both within and between Gunnera populations. It was also found that only one Nostoc strain, or closely related strains, established symbiosis with an individual plant host.

  7. Conserved Chloroplast Open-reading Frame ycf54 Is Required for Activity of the Magnesium Protoporphyrin Monomethylester Oxidative Cyclase in Synechocystis PCC 6803

    Czech Academy of Sciences Publication Activity Database

    Hollingshead, S.; Kopečná, Jana; Jackson, P. J.; Canniffe, D. P.; Davidson, P. A.; Dickman, M. J.; Sobotka, Roman; Hunter, C. N.

    2012-01-01

    Roč. 287, č. 33 (2012), s. 27823-27833 ISSN 0021-9258 R&D Projects: GA ČR GAP501/10/1000; GA MŠk(CZ) ED2.1.00/03.0110 Institutional support: RVO:61388971 Keywords : CHLOROPHYLL ISOCYCLIC RING * RHODOBACTER-SPHAEROIDES * SP PCC-6803 Subject RIV: CE - Biochemistry Impact factor: 4.651, year: 2012

  8. Genetic engineering of Synechocystis PCC6803 for the photoautotrophic production of the sweetener erythritol.

    Science.gov (United States)

    van der Woude, Aniek D; Perez Gallego, Ruth; Vreugdenhil, Angie; Puthan Veetil, Vinod; Chroumpi, Tania; Hellingwerf, Klaas J

    2016-04-08

    Erythritol is a polyol that is used in the food and beverage industry. Due to its non-caloric and non-cariogenic properties, the popularity of this sweetener is increasing. Large scale production of erythritol is currently based on conversion of glucose by selected fungi. In this study, we describe a biotechnological process to produce erythritol from light and CO2, using engineered Synechocystis sp. PCC6803. By functionally expressing codon-optimized genes encoding the erythrose-4-phosphate phosphatase TM1254 and the erythrose reductase Gcy1p, or GLD1, this cyanobacterium can directly convert the Calvin cycle intermediate erythrose-4-phosphate into erythritol via a two-step process and release the polyol sugar in the extracellular medium. Further modifications targeted enzyme expression and pathway intermediates. After several optimization steps, the best strain, SEP024, produced up to 2.1 mM (256 mg/l) erythritol, excreted in the medium.

  9. Optimization of macronutrient kinetics for biomass production in Nostoc calcicola

    Science.gov (United States)

    Aiyer, Subramanian Seshadri C.; Akshai, A.; Kumar, B. G. Prakash; Ramachandran, S.

    2018-04-01

    To assess the feasibility of Allen and Arnon’s (AA) media addition to increase the biomass productivity, (0, 2.5, 5, 7.5 ml of 10x media concentrate - MC) was added to aerated culture every six days, in two separate conditions i.e., single harvest (SH) and continuous harvest (CH) after 15th day. Results show that with addition of 5 ml of MC produced maximum amount of biomass is 1.32 g/L and 2.88 g/L for Sh and CH respectively. These results show that with addition of 5 ml of MC to an aerated culture every six days with continuous biomass harvesting leads to maximum growth of Nostoc calcicola @25°C

  10. Production of cyanopeptolins, anabaenopeptins, and microcystins by the harmful cyanobacteria Anabaena 90 and Microcystis PCC 7806

    NARCIS (Netherlands)

    Tonk, L.; Welker, M.; Huisman, J.; Visser, P.M.

    2009-01-01

    This study investigated the effects of light intensity, temperature, and phosphorus limitation on the peptide production of the cyanobacteria Microcystis PCC 7806 and Anabaena 90. Microcystis PCC 7806 produced two microcystin variants and three cyanopeptolins, whereas Anabaena 90 produced four

  11. Determination of coefficient of thermal expansion effects on Louisiana's PCC pavement design : technical summary report.

    Science.gov (United States)

    2011-12-01

    The coefficient of thermal expansion (CTE) has been widely considered as a fundamental property of : Portland cement concrete (PCC) pavement but has never played an important role in the thickness design : procedure for PCC pavement until recently. I...

  12. Evaluating and optimizing recycled concrete fines in PCC mixtures containing supplementary cementitious materials.

    Science.gov (United States)

    2010-08-01

    Portland cement concrete (PCC) is used throughout transportation infrastructure, for roads as well as bridges : and other structures. One of the most effective ways of making PCC more green is to replace a portion of the : portland cement (the ...

  13. GC/MS analysis of piperidinocyclohexanecarbonitrile (PCC) smoking products

    International Nuclear Information System (INIS)

    Lue, L.P.; Scimeca, J.A.; Thomas, B.F.; Martin, B.R.

    1986-01-01

    Piperidinocyclohexanecarbonitrile (PCC), an intermediate in phencyclidine (PCP) synthesis, is a major contaminant of illicit PCP. Due to the frequent abuse of PCP by smoking, this study was conducted to determine the PCC pyrolysis products delivered in smoke. Marihuana placebo cigarettes were impregnated with 3 H-piperidino- 14 C-cyano-PCC (synthesized in the lab and recrystallized twice, m.p. 67 0 C) and burned under conditions which simulated smoking. Mainstream smoke was passed through glass wool filters and H 2 SO 4 and NaOH traps. Tritium and 14 C were recovered as 83%, and 56%, respectively, of the starting material. Seventy-six percent of the recovered tritium was found in the glass wool trap followed by 13, 7 and 4% in the acid trap, base trap and in the ash/unburned butt, respectively. Seventy-three percent of the recovered 14 C was found in the glass wool filter and 16 and 8% were found in the acid and base traps, respectively. GC/MS analysis revealed the presence of 1-piperidinocyclohexene (30%), PCC (24%), piperidine (7%), and 1-acetyl-piperidine (5%)

  14. Towards PCC for Concurrent and Distributed Systems (Work in Progress)

    Science.gov (United States)

    Henriksen, Anders S.; Filinski, Andrzej

    2009-01-01

    We outline some conceptual challenges in extending the PCC paradigm to a concurrent and distributed setting, and sketch a generalized notion of module correctness based on viewing communication contracts as economic games. The model supports compositional reasoning about modular systems and is meant to apply not only to certification of executable code, but also of organizational workflows.

  15. Transfected parvalbumin alters calcium homeostasis in teratocarcinoma PCC7 cells

    DEFF Research Database (Denmark)

    Müller, B K; Kabos, P; Belhage, B

    1996-01-01

    Indirect evidence supports a protective role of some EF-hand calcium-binding proteins against calcium-induced neurotoxicity. Little is known about how these proteins influence cytosolic calcium levels. After cloning the parvalbumin cDNA into an expression vector, teratocarcinoma cells (PCC7) were...

  16. [Response of Nostoc flageliforme cell to Cu2+, Cr2+ and Pb2+ stress].

    Science.gov (United States)

    Guo, Jinying; Shi, Mingke; Zhao, Yanli; Ren, Guoyan; Yi, Junpeng; Niu, Leilei; Li, Juan

    2013-06-04

    This study aimed to investigate the effects of Cu2+, Cr2+ and Pb2+ stress on Nostoc flagelliforme cell. The response of Nostoc flagelliforme cell was analyzed under the stress. The modified BG11 culture medium containing different heavy metal ions of 0, 0.1, 1.0, 10, 100 mg/L was used to cultivate Nostoc flagelliforme cell at 25 degrees C and light intensity of 80 micromol/(m x s). Electrolyte leakage, the activities of superoxide dismutase, the content of malondialdehyde, proline, soluble protein and trehalose were analyzed. Under 1 - 100 mg/L Cu2+, Cr2+ and Pb2+ stress, electrolyte leakage and malondialdehyde contents in Nostoc flagelliforme cell were higher than those in the control group during heavy metal ions stress. Meanwhile, superoxide dismutase activity increased slightly under 10 mg/L, but was lower afterwards. The contents of proline, soluble protein and trehalose increased under 10 mg/L heavy metal ions stress, while declined under extreme heavy metal ions stress (100 mg/L). Nostoc flagelliforme cell has resistance to low heavy metal ions stress, but is damaged badly under extreme heavy metal ions stress.

  17. Phycobiliprotein accumulation in cyanobacterium Nostoc linckia and modification of antioxidant activity

    Directory of Open Access Journals (Sweden)

    Ana VALUTA

    2015-01-01

    Full Text Available The article deals with iron(III coordination compounds with Schiff bases as ligands and their impact on phycobiliprotein accumulation by cyanobacterium Nostoc linckia. Stimulatory effect depends on the applied dose and in case of three compounds, the concentration 20 mg/L was determined as one with moderate intensity. Lower concentrations resulted in an increase of the phycobiliprotein synthesis. There was found a significant positive correlation between phycobiliprotein content and ABTS (2.2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid assay values displayed by aqueous extracts from Nostoc linckia biomass cultivated in nutrient medium with these coordination compounds. Hence, it is possible to modify the antioxidant activity of Nostoc biomass by applying low concentrations of chemical stimuli.

  18. Effect of IAA on in vitro growth and colonization of Nostoc in plant roots

    Science.gov (United States)

    Hussain, Anwar; Shah, Syed T.; Rahman, Hazir; Irshad, Muhammad; Iqbal, Amjad

    2015-01-01

    Nostoc is widely known for its ability to fix atmospheric nitrogen and the establishment of symbiotic relationship with a wide range of plants from various taxonomic groups. Several strains of Nostoc produce phytohormones that promote growth of its plant partners. Nostoc OS-1 was therefore selected for study because of the presence of putative ipdC gene that encodes a key enzyme to produce Indole-3-acetic acid (IAA). The results indicated that both cellular and released IAA was found high with increasing incubation time and reached to a peak value (i.e., 21 pmol mg-1ch-a) on the third week as determined by UPLC-ESI-MS/MS. Also the Nostoc OS-1 strain efficiently colonized the roots and promoted the growth of rice as well as wheat under axenic conditions and induced ipdC gene that suggested the possible involvement of IAA in these phenotypes. To confirm the impact of IAA on root colonization efficiency and plant promoting phenotypes of Nostoc OS-1, an ipdC knockout mutant was generated by homologous recombinant method. The amount of releasing IAA, in vitro growth, root colonization, and plant promoting efficiency of the ipdC knockout mutant was observed significantly lower than wild type strain under axenic conditions. Importantly, these phenotypes were restored to wild-type levels when the ipdC knockout mutant was complemented with wild type ipdC gene. These results together suggested that ipdC and/or synthesized IAA of Nostoc OS-1 is required for its efficient root colonization and plant promoting activity. PMID:25699072

  19. COMPONENŢA AMINOACIZILOR DIN BIOMASA ALGEI CIANOFITE NOSTOC GELATINOSUM (SCHOUSB ELENK.

    Directory of Open Access Journals (Sweden)

    Sergiu DOBROJAN

    2016-02-01

    Full Text Available În articol este analizată componenţa aminoacizilor din biomasa algei cianofite Nostoc gelatinosum (Schousb Elenk. colectate de pe mediul nutritiv Drew. Din grupa aminoacizilor esenţiali în biomasa algei Nostoc gelatinosum (Schousb Elenk. se conţine în cantităţi majore leucina (2,142±0,06 mg/100 mg, treonina (1,188±0,02 mg/100 mg şi valina (1,085±0,03 mg/100 mg. Din aminoacizii neesenţiali predomină acidul aspartic (4,523±0,11 mg/100 mg şi acidul glutamic (2,774±0,07 mg/100 mg. Alga Nostoc gelatinosum (Schousb Elenk. are un conţinut bogat de aminoacizi şi poate servi ca sursă pentru vaste domenii de aplicare.THE AMINO ACIDS COMPOSITION OF THE BLUE-GREEN ALGANOSTOC GELATINOSUM (SCHOUSB ELENKThis article presents the amino acid content of the blue-green alga Nostoc gelatinosum (Schousb Elenk. collected from the Drew nutritive medium. From the essential amino group in Nostoc gelatinosum (Schousb Elenk. biomass, major amounts are contained leucine (2,142 ± 0,06 mg/100 mg, threonine (1,188 ± 0,02 mg/100 mg and valine (1,085 ± 0,03 mg/100 mg. From nonessential amino acids predominates aspartic acid (4,523 ± 0,11 mg/100 mg and glutamic acid (2,774 ± 0.07 mg/100 mg. The alga Nostoc gelatinosum (Schousb Elenk. has a high content of amino acids and can serve as a source for vast areas of application. 

  20. Biochemical Validation of the Glyoxylate Cycle in the Cyanobacterium Chlorogloeopsis fritschii Strain PCC 9212.

    Science.gov (United States)

    Zhang, Shuyi; Bryant, Donald A

    2015-05-29

    Cyanobacteria are important photoautotrophic bacteria with extensive but variable metabolic capacities. The existence of the glyoxylate cycle, a variant of the TCA cycle, is still poorly documented in cyanobacteria. Previous studies reported the activities of isocitrate lyase and malate synthase, the key enzymes of the glyoxylate cycle in some cyanobacteria, but other studies concluded that these enzymes are missing. In this study the genes encoding isocitrate lyase and malate synthase from Chlorogloeopsis fritschii PCC 9212 were identified, and the recombinant enzymes were biochemically characterized. Consistent with the presence of the enzymes of the glyoxylate cycle, C. fritschii could assimilate acetate under both light and dark growth conditions. Transcript abundances for isocitrate lyase and malate synthase increased, and C. fritschii grew faster, when the growth medium was supplemented with acetate. Adding acetate to the growth medium also increased the yield of poly-3-hydroxybutyrate. When the genes encoding isocitrate lyase and malate synthase were expressed in Synechococcus sp. PCC 7002, the acetate assimilation capacity of the resulting strain was greater than that of wild type. Database searches showed that the genes for the glyoxylate cycle exist in only a few other cyanobacteria, all of which are able to fix nitrogen. This study demonstrates that the glyoxylate cycle exists in a few cyanobacteria, and that this pathway plays an important role in the assimilation of acetate for growth in one of those organisms. The glyoxylate cycle might play a role in coordinating carbon and nitrogen metabolism under conditions of nitrogen fixation. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  1. Interactions of copper and lead with Nostoc muscorum

    Energy Technology Data Exchange (ETDEWEB)

    Schecher, W.D.; Driscoll, C.T.

    1985-01-01

    Experiments were conducted to evaluate the effects of cell concentration, time of exposure, cellular activity and solution chemistry, on Pb (10/sup -6/ M) and Cu (10/sup -5/ M) uptake by the alga Nostoc muscorum. Surface equilibrium, with respect to aqueous metal levels, was established within an equilibration period of 8 h and maximum metal removal was observed in the pH range of 7.5 to 8.0. The observed removal of Cu and Pb from solution was similar to adsorption observed for inorganic surfaces at pH values less than 8.0. Removal of metallic ions decreased at pH values greater than 8.0 which was thought to be due to aqueous complexation with organic extracellular material. The extent to which the cells were able to remove trace metals from solution in the presence of citrate, sulfate, and Ca ion (10/sup -3/ M) was also evaluated. Additions of citrate and Ca ion mitigated metal uptake by algal suspensions. The presence of sulfate resulted in a reduction of Cu removal below pH values of 5.6 but enhanced the removal of Pb over the entire pH range. The chemical equilibrium model MINEQL was utilized to compare theoretical and observed phenomena so that possible mechanisms for metal-cell interactions could be assessed.

  2. Active accumulation of internal DIC pools reduces transport limitation in large colonies of Nostoc pruniforme

    DEFF Research Database (Denmark)

    Raun, Ane-Marie Løvendahl; Borum, Jens; Jensen, Kaj Sand

    2009-01-01

    Nostoc pruniforme is a freshwater cyanobacterium forming large spherical colonies of up to several centimeters in diameter. The size and shape result in low surface area to volume (SA/V) ratios that potentially put severe constraints on resource acquisition. In the present study we have specifica......Nostoc pruniforme is a freshwater cyanobacterium forming large spherical colonies of up to several centimeters in diameter. The size and shape result in low surface area to volume (SA/V) ratios that potentially put severe constraints on resource acquisition. In the present study we have...

  3. Microbial Carbonate Precipitation by Synechococcus PCC8806, LS0519 and Synechocystis PCC6803 on Concrete Surfaces and in Low Saturation Solution

    Science.gov (United States)

    Zhu, T.; Lin, Y.; Dittrich, M.

    2015-12-01

    Microbial carbonate precipitation (MCP) by cyanobacteria has been recognized in a variety of environment such as freshwater, marine, cave, and even desert. Recently, their calcification potential has been tested in an emerging technology-- bioconcrete. This study is to explore the calcification by three cyanobacteria strains under different environmental conditions. Experiment A was carried out in 2mM NaHCO3 and 5mM CaCl2, with a cell concentration of 107 cells L-1. In experiment B, one side of the concrete surface was treated with bacteria and then immersed in the solution containing 0.4 mM NaHCO3 and 300 mM CaCl2. In experiment A, the pH of the abiotic condition remained constant around 8.55, while that of biotic conditions increased by 0.15 units in the presence of LS0519, and by 0.3 units in the presence of PCC8806 or PCC6803 within 8 hours. Over a period of 30 hours, PCC8806, LS0519 and PCC6803 removed 0.1, 0.12 and 0.2 mM calcium from the solution respectively. After 30 hours, the alkalinity of the solution decreased by 30 mg/L, 10 mg/L and 5 mg/L respectively in the presence of PCC6803, LS0519 and PCC8806. Under scanning electron microscopy (SEM), no precipitate was found in the abiotic condition, while calcium carbonate was associated by all the three strains. Among them, PCC6803 precipitated more carbonates. In experiment B, LS0519 and PCC8806 increased the pH with a value of 0.25, while PCC6803 increased the pH by 0.33 units. SEM shows LS0519 was less likely attached to the concrete surface. Neither did the precipitates on concrete surface differ from that in the abiotic condition. In comparison, PCC8806 and PCC6803 were closely associated with 8-μm porous precipitates. Cells were either found enclosed in precipitates or connecting two precipitates. In conclusion, all the three strains triggered the calcium carbonate precipitation. LS0519 has a little impact on the carbonate precipitation in the solution, but negligent influence on the concrete surface

  4. Nitrogenase activity (acetylene reduction activity) and diversity of six soil Nostoc strains

    Czech Academy of Sciences Publication Activity Database

    Hrouzek, P.; Šimek, Miloslav; Komárek, Jiří

    2003-01-01

    Roč. 108, - (2003), s. 87-101 ISSN 0342-1120 R&D Projects: GA AV ČR KSK6005114 Institutional research plan: CEZ:AV0Z6066911 Keywords : cyanobacteria * Nostoc * morphology Subject RIV: EH - Ecology, Behaviour

  5. Genus .i.Nostoc./i. - a source of novel trypsin inhibitors

    Czech Academy of Sciences Publication Activity Database

    Drápalová, P.; Štys, D.; Lukešová, Alena; Kopecký, Jiří

    2008-01-01

    Roč. 127, č. 1 (2008), s. 61-82 ISSN 1864-1318 R&D Projects: GA MŠk ME 874 Institutional research plan: CEZ:AV0Z60660521; CEZ:AV0Z50200510 Keywords : cyanobacteria * LC/MS * Nostoc Subject RIV: EH - Ecology, Behaviour

  6. Unprecedented slow growth and mortality of the rare colonial cyanobacterium, Nostoc zetterstedtii, in oligotrophic lakes

    DEFF Research Database (Denmark)

    Jensen, Kaj Sand; Møller, Claus Lindskov

    2011-01-01

    Centimeter-large colonies of Nostoc zetterstedtii from a Swedish oligotrophic lake had the lowest growth and mortality rates of any studied temperate macrophyte. Annual growth rates at two shallow sites averaged 0.57– 0.73 3 1023 d21, corresponding to doubling times of colony dry weight in 2...

  7. Effect of a combination of two rice herbicides on the cyanobacterium, Nostoc spongiaeforme

    Science.gov (United States)

    Cyanobacteria grow in California rice fields where they form large mats that may smoother seedlings or cause them to dislodge, resulting in yield loss. The most troublesome species is Nostoc spongiaeforme. It is very difficult to control using currently accepted methods, i.e., aerial applications of...

  8. Morphological characterization and molecular fingerprinting of Nostoc strains by multiplex RAPD.

    Science.gov (United States)

    Hillol, Chakdar; Pabbi, Sunil

    2012-01-01

    Morphological parameters studied for the twenty selected Nostoc strains were mostly found to be consistent with the earlier reports. But the shape of akinetes observed in this study was a little deviation from the existing descriptions and heterocyst frequency was also found to be different in different strains in spite of growing in the same nitrogen free media. Multiplex RAPD produced reproducible and completely polymorphic amplification profiles for all the strains including some strain specific unique bands which are intended to be useful for identification of those strains. At least one to a maximum of two unique bands was produced by different dual primer combinations. For ten strains out of twenty, strain specific bands were found to be generated. Cluster analysis revealed a vast heterogeneity among these Nostoc strains and no specific clustering based on geographical origin was found except a few strains. It was also observed that morphological data may not necessarily correspond to the genetic data in most of the cases. CCC92 (Nostoc muscorum) and CCC48 (Nostoc punctiforme) showed a high degree of similarity which was well supported by high bootstrap value. The level of similarity of the strains ranged from 0.15 to 0.94. Cluster analysis based on multiplex RAPD showed a good fit revealing the discriminatory power of this technique.

  9. The cyanobiont in an Azolla fern is neither Anabaena nor Nostoc.

    Science.gov (United States)

    Baker, Judith A; Entsch, Barrie; McKay, David B

    2003-12-05

    The cyanobacterial symbionts in the fern Azolla have generally been ascribed to either the Anabaena or Nostoc genera. By using comparisons of the sequences of the phycocyanin intergenic spacer and a fragment of the 16S rRNA, we found that the cyanobiont from an Azolla belongs to neither of these genera.

  10. Single-cell confocal spectrometry of a filamentous cyanobacterium Nostoc at room and cryogenic temperature. Diversity and differentiation of pigment systems in 311 cells.

    Science.gov (United States)

    Sugiura, Kana; Itoh, Shigeru

    2012-08-01

    The fluorescence spectrum at 298 and 40 K and the absorption spectrum at 298 K of each cell of the filamentous cyanobacterium Nostoc sp. was measured by single-cell confocal laser spectroscopy to study the differentiation of cell pigments. The fluorescence spectra of vegetative (veg) and heterocyst (het) cells of Nostoc formed separate groups with low and high PSII to PSI ratios, respectively. The fluorescence spectra of het cells at 40 K still contained typical PSII bands. The PSII/PSI ratio estimated for the veg cells varied between 0.4 and 1.2, while that of het cells varied between 0 and 0.22 even in the same culture. The PSII/PSI ratios of veg cells resembled each other more closely in the same filament. 'pro-het' cells, which started to differentiate into het cells, were identified from the small but specific difference in the PSII/PSI ratio. The allophycocyanin (APC)/PSII ratio was almost constant in both veg and het cells, indicating their tight couplings. Phycocyanin (PC) showed higher fluorescence in most het cells, suggesting the uncoupling from PSII. Veg cells seem to vary their PSI contents to give different PSII/PSI ratios even in the same culture, and to suppress the synthesis of PSII, APC and PC to differentiate into het cells. APC and PC are gradually liberated from membranes in het cells with the uncoupling from PSII. Single-cell spectrometry will be useful to study the differentiation of intrinsic pigments of cells and chloroplasts, and to select microbes from natural environments.

  11. INFLUENCE OF NOSTOC VAUCHER EX BORNET ET FLAHAULT STRAINS ON GROWTH AND DEVELOPMENT OF PISUM SATIVUM L.

    Directory of Open Access Journals (Sweden)

    Maltsev Ye. I.

    2015-12-01

    Full Text Available We investigated the positive impact of cultures cyanobacteria genus Nostoc Vaucher ex Bornet et Flahault on growth and development of higher plants as an example Pisum sativum L. All the Nostoc species have a positive effect on the viability, germination energy, and biometric characteristics of Pisum sativum. The greatest positive influence was registered for N. entophytum Born. et. Flah. and N. linckia (Roth Bornetet Flahault f. linckia.

  12. Synthesis of Hydroxyapatite using Precipitated Calcium Carbonate (PCC) from Limestones

    Science.gov (United States)

    Wardhani, Sri; Isnaini Azkiya, Noor; Triandi Tjahjanto, Rachmat

    2018-01-01

    Hydroxyapatite (HAp) is a material that widely applied in bone and teeth implant due to its biocompatibility and bioactivity. This material can be prepared from PCC by precipitation method using CaO and H3PO4 in ethanol. In this work, variations of phosphoric acid amount and aging time were investigated. The synthesized HAp was characterized by FT-IR, AAS, UV-Vis Spectrophotometer, PSA, SEM, and powder XRD. The results showed that the high concentration of calcium in PCC gives better yields in which PCC obtained from carbonation method has higher yield than that of caustic soda method. The determination of optimum phosphoric acid addition based on targeted Ca/P ratio (1.67) from HAp was obtained on the addition of 0.1271 mol phosphoric acid with Ca/P ratio of 1.66. The aging time gave significant effect to the particle size of synthesised HAp. The smallest particle size was obtained in aging time for 48 hours as high as 49.25 μm. FTIR spectra of the synthesized HAp show the presence of hydroxyl (-OH) group at 3438.8 cm-1, PO4 3- at 557.39 and 1035.7 cm-1, and CaO at 1413.72 cm-1. The synthesized HAp forms agglomeration solid based on the SEM analysis. The powder XRD data shows three highest peaks at 2θ i.e. 27.8296; 31.1037; and 34.3578 which corresponds to β-TCP (tricalcium phosphate) in accordance with JCPDS no.09-0169. The characteristic 2θ peak of hydroxyapatite with low intensity is observed from the synthesized HAp refer to the JCPDS data no. 09-0432.

  13. Molecular and phylogenetic characterization of two species of the genus Nostoc (Cyanobacteria based on the cpcB-IGS-cpcA locus of the phycocyanin operon

    Directory of Open Access Journals (Sweden)

    IVANKA TENEVA

    2012-01-01

    Full Text Available Traditionally, the taxonomy of the genus Nostoc is based on morphological and physiological characters. The extreme morphological variability of the Nostoc species, due to their life cycle and environmental conditions, hampers the correct identification of the individual species. This is also one of the reasons for the disputed taxonomic positions and relationships between the genera Anabaena–Aphanizomenon as well as between Anabaena–Nostoc. Therefore, it is necessary to use additional markers for development of a polyphasic classification system of order Nostocales. In light of this, we here present the first molecular and phy-logenetic characterization of two species of the genus Nostoc (Nostoc linckia and Nostoc punctiforme based on the cpcB-IGS-cpcA locus of the phycocyanin oper-on. The phylogenetic position of these two species within order Nostocales as well as within division Cyanobacteria has been determined. Our results indicate that genus Nostoc is heterogeneous. Analysis of the IGS region between cpcB and cpcA showed that Nostoc and Anabaena are distinct genera. Reported molecular and phylogenetic data will be useful to solve other problematic points in the tax-onomy of genera Aphanizomenon, Anabaena and Nostoc.

  14. Evaluation of the MIT-Scan-T2 for non-destructive PCC pavement thickness determination.

    Science.gov (United States)

    2008-07-01

    The MIT-Scan-T2 device is marketed as a non-destructive way to determine pavement thickness on both : HMA and PCC pavements. PCC pavement thickness determination is an important incentivedisincentive : measurement for the Iowa DOT and contractors. Th...

  15. Evaluation of MIT-SCAN-T2 for Thickness Quality Control for PCC and HMA Pavements

    Science.gov (United States)

    2018-02-01

    Thickness is currently a pay item for PCC pavements and a quality control item for both PCC and HMA pavements. A change in pavement thickness of 0.5 in. can result in a change of multiple years of service. Current thickness measurements are performed...

  16. Role of enzymatic and non enzymatic antioxidant in ameliorating salinity induced damage in nostoc muscorum

    International Nuclear Information System (INIS)

    Hend, A.; Abeer, A.; Allah, A.

    2015-01-01

    Presence of high salt concentration in the growth medium adversely affected the plant growth and productivity by altering its metabolic activities. Experiments were conducted on cyanobacteriaum Nostoc muscorum grown in nitrogen free medium supplemented with 250 mM NaCl to evaluate the salt stress induced changes in growth, antioxidants and lipid composition. Salt stress significantly reduced the growth and physio-biochemical attributes. Salt stress increased malonaldehyde content thereby causing alterations in the lipid fraction. Significant reduction in polyunsaturated fatty acids including phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidylinositol (PI) and phosphatidylserine (PS) was observed. Where as diacylglycerol, sterol ester and non-esterified fatty acids were increased. Activities of antioxidant enzymes and contents of non-enzymatic antioxidants including glutathione enhanced due to salt stress. An increase in accumulation of proline was also observed. Hence increased activity of antioxidants and altered fatty acid composition was observed in salt stressed Nostoc muscorum. (author)

  17. Antioxidative activity of ethanol extracts from Spirulina platensis and Nostoc linckia measured by various methods

    OpenAIRE

    Liliana CEPOI; Ludmila RUDI; Vera MISCU; Angela COJOCARI; Tatiana CHIRIAC; Daniela SADOVNIC

    2009-01-01

    The goal of this work is to determine the level of antioxidative activity of various ethanol extracts from Spirulina platensis and Nostoc linckia biomass, and also to demonstrate the possibility to select suitable methods for evaluation of these characteristics. The methods for determination of antioxidative activity were selected concerning their possible use for complex preparations: phosphomolybdenum method for evaluation of antioxidant capacity (PMRC), radical-scavenging activity by DPPH ...

  18. Multiple Roles of Soluble Sugars in the Establishment of Gunnera-Nostoc Endosymbiosis1[OA

    Science.gov (United States)

    Khamar, Hima J.; Breathwaite, Erick K.; Prasse, Christine E.; Fraley, Elizabeth R.; Secor, Craig R.; Chibane, Fairouz L.; Elhai, Jeff; Chiu, Wan-Ling

    2010-01-01

    Gunnera plants have the unique ability to form endosymbioses with N2-fixing cyanobacteria, primarily Nostoc. Cyanobacteria enter Gunnera through transiently active mucilage-secreting glands on stems. We took advantage of the nitrogen (N)-limitation-induced gland development in Gunnera manicata to identify factors that may enable plant tissue to attract and maintain cyanobacteria colonies. Cortical cells in stems of N-stressed Gunnera plants were found to accumulate a copious amount of starch, while starch in the neighboring mature glands was nearly undetectable. Instead, mature glands accumulated millimolar concentrations of glucose (Glc) and fructose (Fru). Successful colonization by Nostoc drastically reduced sugar accumulation in the surrounding tissue. Consistent with the abundance of Glc and Fru in the gland prior to Nostoc colonization, genes encoding key enzymes for sucrose and starch hydrolysis (e.g. cell wall invertase, α-amylase, and starch phosphorylase) were expressed at higher levels in stem segments with glands than those without. In contrast, soluble sugars were barely detectable in mucilage freshly secreted from glands. Different sugars affected Nostoc’s ability to differentiate motile hormogonia in a manner consistent with their locations. Galactose and arabinose, the predominant constituents of polysaccharides in the mucilage, had little or no inhibitory effect on hormogonia differentiation. On the other hand, soluble sugars that accumulated in gland tissue, namely sucrose, Glc, and Fru, inhibited hormogonia differentiation and enhanced vegetative growth. Results from this study suggest that, in an N-limited environment, mature Gunnera stem glands may employ different soluble sugars to attract Nostoc and, once the cyanobacteria are internalized, to maintain them in the N2-fixing vegetative state. PMID:20833727

  19. Impact of warming on Nostoc colonies (Cyanobacteria) in a wet hummock meadow, Spitsbergen

    Czech Academy of Sciences Publication Activity Database

    Elster, Josef; Kvíderová, Jana; Hájek, Tomáš; Láska, K.; Šimek, Miloslav

    2012-01-01

    Roč. 33, č. 4 (2012), 395-420 ISSN 0138-0338 R&D Projects: GA MŠk ME 934; GA MŠk LA341 Institutional research plan: CEZ:AV0Z60050516; CEZ:AV0Z60660521 Institutional support: RVO:67985939 ; RVO:60077344 Keywords : global climate change * Nostoc * OTC Subject RIV: EH - Ecology, Behaviour Impact factor: 0.745, year: 2012

  20. Structural organisation of phycobilisomes from Synechocystis sp strain PCC6803 and their interaction with the membrane

    NARCIS (Netherlands)

    Arteni, Ana A.; Ajlani, Ghada; Boekema, Egbert J.

    In cyanobacteria, the harvesting of light energy for photosynthesis is mainly carried out by the phycobilisome a giant, multi-subunit pigment-protein complex. This complex is composed of heterodimeric phycobiliproteins that are assembled with the aid of linker polypeptides such that light absorption

  1. Protein Network Signatures Associated with Exogenous Biofuels Treatments in Cyanobacterium Synechocystis sp. PCC 6803

    International Nuclear Information System (INIS)

    Pei, Guangsheng; Chen, Lei; Wang, Jiangxin; Qiao, Jianjun; Zhang, Weiwen

    2014-01-01

    Although recognized as a promising microbial cell factory for producing biofuels, current productivity in cyanobacterial systems is low. To make the processes economically feasible, one of the hurdles, which need to be overcome is the low tolerance of hosts to toxic biofuels. Meanwhile, little information is available regarding the cellular responses to biofuels stress in cyanobacteria, which makes it challenging for tolerance engineering. Using large proteomic datasets of Synechocystis under various biofuels stress and environmental perturbation, a protein co-expression network was first constructed and then combined with the experimentally determined protein–protein interaction network. Proteins with statistically higher topological overlap in the integrated network were identified as common responsive proteins to both biofuels stress and environmental perturbations. In addition, a weighted gene co-expression network analysis was performed to distinguish unique responses to biofuels from those to environmental perturbations and to uncover metabolic modules and proteins uniquely associated with biofuels stress. The results showed that biofuel-specific proteins and modules were enriched in several functional categories, including photosynthesis, carbon fixation, and amino acid metabolism, which may represent potential key signatures for biofuels stress responses in Synechocystis. Network-based analysis allowed determination of the responses specifically related to biofuels stress, and the results constituted an important knowledge foundation for tolerance engineering against biofuels in Synechocystis.

  2. Cryo-imaging of photosystems and phycobilisomes in Anabaena sp PCC 7120 cells

    Czech Academy of Sciences Publication Activity Database

    Steinbach, Gabor; Schubert, F.; Kaňa, Radek

    2015-01-01

    Roč. 152, NOV 2015 (2015), s. 395-399 ISSN 1011-1344 R&D Projects: GA ČR GAP501/12/0304; GA MŠk EE2.3.30.0059; GA MŠk ED2.1.00/03.0110; GA MŠk LO1416 Institutional support: RVO:61388971 Keywords : Confocal microscopy * Cryogenic microscopy * Cyanobacteria Subject RIV: CE - Biochemistry Impact factor: 3.035, year: 2015

  3. Expression, purification and preliminary crystallization study of RpaC protein from Synechocystis sp PCC6803

    Czech Academy of Sciences Publication Activity Database

    Cséfalvay, Eva; Lapkouski, Mikalai; Komárek, Ondřej

    2009-01-01

    Roč. 3, č. 47 (2009), s. 355-362 ISSN 0300-3604 R&D Projects: GA ČR GA206/07/0917 Institutional research plan: CEZ:AV0Z60870520 Keywords : affinity chromatography * photosystem * phycobilisome – PSII supercomplex Subject RIV: CE - Biochemistry Impact factor: 1.072, year: 2009

  4. Protein Network Signatures Associated with Exogenous Biofuels Treatments in Cyanobacterium Synechocystis sp. PCC 6803

    Energy Technology Data Exchange (ETDEWEB)

    Pei, Guangsheng; Chen, Lei; Wang, Jiangxin; Qiao, Jianjun, E-mail: jianjunq@tju.edu.cn; Zhang, Weiwen, E-mail: jianjunq@tju.edu.cn [Laboratory of Synthetic Microbiology, School of Chemical Engineering and Technology, Tianjin University, Tianjin (China); Key Laboratory of Systems Bioengineering, Ministry of Education of China, Tianjin (China); SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering, Tianjin (China)

    2014-11-03

    Although recognized as a promising microbial cell factory for producing biofuels, current productivity in cyanobacterial systems is low. To make the processes economically feasible, one of the hurdles, which need to be overcome is the low tolerance of hosts to toxic biofuels. Meanwhile, little information is available regarding the cellular responses to biofuels stress in cyanobacteria, which makes it challenging for tolerance engineering. Using large proteomic datasets of Synechocystis under various biofuels stress and environmental perturbation, a protein co-expression network was first constructed and then combined with the experimentally determined protein–protein interaction network. Proteins with statistically higher topological overlap in the integrated network were identified as common responsive proteins to both biofuels stress and environmental perturbations. In addition, a weighted gene co-expression network analysis was performed to distinguish unique responses to biofuels from those to environmental perturbations and to uncover metabolic modules and proteins uniquely associated with biofuels stress. The results showed that biofuel-specific proteins and modules were enriched in several functional categories, including photosynthesis, carbon fixation, and amino acid metabolism, which may represent potential key signatures for biofuels stress responses in Synechocystis. Network-based analysis allowed determination of the responses specifically related to biofuels stress, and the results constituted an important knowledge foundation for tolerance engineering against biofuels in Synechocystis.

  5. Structural and mutational analysis of band 7 proteins in the cyanobacterium Synechocystis sp. strain PCC 6803

    Czech Academy of Sciences Publication Activity Database

    Boehm, M.; Nield, J.; Zhang, P.; Aro, E.-M.; Komenda, Josef; Nixon, P. J.

    2009-01-01

    Roč. 191, č. 20 (2009), s. 6425-6435 ISSN 0021-9193 R&D Projects: GA AV ČR IAA400200801 Institutional research plan: CEZ:AV0Z50200510 Keywords : BLUE NATIVE ELECTROPHORESIS * PHOTOSYSTEM-II COMPLEX * COLI PLASMA-MEMBRANE Subject RIV: EE - Microbiology, Virology Impact factor: 3.940, year: 2009

  6. Mechanisms of High Temperature Resistance of Synechocystis sp. PCC 6803: An Impact of Histidine Kinase 34

    Czech Academy of Sciences Publication Activity Database

    Červený, Jan; Sinětova, M. A.; Zavřel, Tomáš; Los, D. A.

    2015-01-01

    Roč. 5, č. 1 (2015), s. 676-699 ISSN 2075-1729 R&D Projects: GA MŠk(CZ) ED1.1.00/02.0073; GA MŠk(CZ) EE2.3.20.0256; GA MŠk(CZ) LO1415 Institutional support: RVO:67179843 Keywords : photosynthesis * pigments * ultrastructure * heat stress proteins * photobioreactor * cyanobacteria Subject RIV: EH - Ecology, Behaviour

  7. Nostoc sphaeroides Kützing, an excellent candidate producer for CELSS

    Science.gov (United States)

    Hao, Zongjie; Li, Dunhai; Li, Yanhui; Wang, Zhicong; Xiao, Yuan; Wang, Gaohong; Liu, Yongding; Hu, Chunxiang; Liu, Qifang

    2011-11-01

    Some phytoplankton can be regarded as possible candidates in the establishment of Controlled Ecological Life Support System (CELSS) for some intrinsic characteristics, the first characteristic is that they should grow rapidly, secondly, they should be able to endure some stress factors and develop some corresponding adaptive strategies; also it is very important that they could provide food rich in nutritious protein and vitamins for the crew; the last but not the least is they can also fulfill the other main functions of CELSS, including supplying oxygen, removing carbon dioxide and recycling the metabolic waste. According to these characteristics, Nostoc sphaeroides, a potential healthy food in China, was selected as the potential producer in CELSS. It was found that the oxygen average evolution rate of this algae is about 150 μmol O 2 mg -1 h -1, and the size of them are ranged from 2 to 20 mm. Also it can be cultured with high population density, which indicated that the potential productivity of Nostoc sphaeroides is higher than other algae in limited volume. We measured the nutrient contents of the cyanobacterium and concluded it was a good food for the crew. Based on above advantages, Nostoc sphaeroides was assumed to a suitable phytoplankton for the establishment of Controlled Ecological Life Support System. We plan to develop suitable bioreactor with the cyanobacterium for supplying oxygen and food in future space missions.

  8. Physicochemical Characterization and Functional Analysis of the Polysaccharide from the Edible Microalga Nostoc sphaeroides

    Directory of Open Access Journals (Sweden)

    Haifeng Li

    2018-02-01

    Full Text Available Nostoc colonies have been used as food and medicine for centuries, and their main supporting matrix is polysaccharides, which help Nostoc cells resist various environmental stresses including oxidative stress. Here we isolated a polysaccharide, nostoglycan, from cultured Nostoc sphaeroides colonies and determined its physicochemical properties, which revealed a characteristic infrared absorption spectrum typical of polysaccharides and an amorphous morphology with rough surfaces. We also show that nostoglycan has strong moisture absorption and retention capacities and a high relative viscosity. Using Caenorhabditis elegans models, we then demonstrate that nostoglycan is capable of improving overall survival rate of the animals under increased oxidative stress caused by paraquat. Nostoglycan also reduces reactive oxygen species level, inhibits protein carbonyl formation and lipid peroxidation, and increases activities of superoxide dismutase and catalase in paraquat-exposed nematodes. As oxidative stress may drive tumor progression, we further demonstrate that nostoglycan can suppress the proliferation of several types of tumor cells and induce apoptosis of human lung adenocarcinoma A549 cells via caspase-3 activation. Together, our results yield important information on the physicochemical characteristics and demonstrate the antioxidant and anti-proliferative functions of nostoglycan, and thus provide an insight into its potential in food and health industries.

  9. Diversity of endosymbiotic Nostoc in Gunnera magellanica from Tierra del Fuego, Chile [corrected].

    Science.gov (United States)

    Fernández-Martínez, M A; de Los Ríos, A; Sancho, L G; Pérez-Ortega, S

    2013-08-01

    Global warming is causing ice retreat in glaciers worldwide, most visibly over the last few decades in some areas of the planet. One of the most affected areas is the region of Tierra del Fuego (southern South America). Vascular plant recolonisation of recently deglaciated areas in this region is initiated by Gunnera magellanica, which forms symbiotic associations with the cyanobacterial genus Nostoc, a trait that likely confers advantages in this colonisation process. This symbiotic association in the genus Gunnera is notable as it represents the only known symbiotic relationship between angiosperms and cyanobacteria. The aim of this work was to study the genetic diversity of the Nostoc symbionts in Gunnera at three different, nested scale levels: specimen, population and region. Three different genomic regions were examined in the study: a fragment of the small subunit ribosomal RNA gene (16S), the RuBisCO large subunit gene coupled with its promoter sequence and a chaperon-like protein (rbcLX) and the ribosomal internal transcribed spacer (ITS) region. The identity of Nostoc as the symbiont was confirmed in all the infected rhizome tissue analysed. Strains isolated in the present study were closely related to strains known to form symbioses with other organisms, such as lichen-forming fungi or bryophytes. We found 12 unique haplotypes in the 16S rRNA (small subunit) region analysis, 19 unique haplotypes in the ITS region analysis and 57 in the RuBisCO proteins region (rbcLX). No genetic variability was found among Nostoc symbionts within a single host plant while Nostoc populations among different host plants within a given sampling site revealed major differences. Noteworthy, interpopulation variation was also shown between recently deglaciated soils and more ancient ones, between eastern and western sites and between northern and southern slopes of Cordillera Darwin. The cell structure of the symbiotic relationship was observed with low-temperature scanning

  10. POTENŢIALUL NUTRIŢIONAL AL BIOMASEI ALGALE DE NOSTOC FLAGELLIFORME (BERK ET CURT ELENK.

    Directory of Open Access Journals (Sweden)

    Irina STRATULAT

    2016-05-01

    Full Text Available În prezentul studiu este prezentată componenţa biochimică a biomasei algei cianofite Nostoc flagelli­forme (Berk et Curt Elenk. cultivată în condiţii de laborator pe mediul mineral Z-8. Biomasa algei conţine: Mn – 683,9 mg/kg, Zn – 131,5 mg/kg, Pb – <10 mg/kg, Fe – 1,09 g/kg, Na+ – 2,10 g/kg, K+ – 8,01 g/kg, Mg 2+ – 5,40 g/kg, Ca2+ – 7,94 g/kg. În componenţa aminoacizilor predomină: acidul glutamic – 3,8012 mg/100 mg, acidul aspartic – 2,7838 mg/100 mg, leucina – 2,4651 mg/100 mg, alanina – 2,3513 mg/100 mg. Conform acestor rezultate, biomasa algei cianofite Nostoc flagelliforme poate fi utilizată ca sursă importantă de supli­mente alimentare. THE NUTRITIONAL POTENTIAL OF ALGAL BIOMASS OF NOSTOC FLAGELLIFORME (BERK ET CURT ELENK. In this study is presented the biochemical components of algal biomass Nostoc flagelliforme (Berk et Curt Elenk. cultivated in laboratory conditions in Z-8 medium. The algal biomass has the following chemical content: Mn – 683,9 mg/kg, Zn – 131,5 mg/kg, Pb – <10 mg/kg, Fe – 1,09 g/kg, Na+ - 2,10 g/kg, K+ – 8,01 g/kg, Mg2+ – 5,40 g/kg, Ca2+ – 7,94 g/kg. The highest amount of amino acids are specific for: glutamic acid – 3,8012 mg/100 mg, aspartic acid – 2,7838 mg/100 mg, leucine – 2,4651 mg/100 mg, alanine – 2,3513 mg/100 mg. According to these results the blue-green alga Nostoc flagelliforme can be considered an important potential source of nutritional supplements. 

  11. Drug-induced premature chromosome condensation (PCC) protocols: cytogenetic approaches in mitotic chromosome and interphase chromatin.

    Science.gov (United States)

    Gotoh, Eisuke

    2015-01-01

    Chromosome analysis is a fundamental technique which is used in wide areas of cytogenetic study including karyotyping species, hereditary diseases diagnosis, or chromosome biology study. Chromosomes are usually prepared from mitotic cells arrested by colcemid block protocol. However, obtaining mitotic chromosomes is often hampered under several circumstances. As a result, cytogenetic analysis will be sometimes difficult or even impossible in such cases. Premature chromosome condensation (PCC) (see Note 1) is an alternative method that has proved to be a unique and useful way in chromosome analysis. Former, PCC has been achieved following cell fusion method (cell-fusion PCC) mediated either by fusogenic viruses (e.g., Sendai virus) or cell fusion chemicals (e.g., polyethylene glycol), but the cell fusion PCC has several drawbacks. The novel drug-induced PCC using protein phosphatase inhibitors was introduced about 20 years ago. This method is much simpler and easier even than the conventional mitotic chromosome preparation protocol use with colcemid block and furthermore obtained PCC index (equivalent to mitotic index for metaphase chromosome) is usually much higher than colcemid block method. Moreover, this method allows the interphase chromatin to be condensed to visualize like mitotic chromosomes. Therefore drug-induced PCC has opened the way for chromosome analysis not only in metaphase chromosomes but also in interphase chromatin. The drug-induced PCC has thus proven the usefulness in cytogenetics and other cell biology fields. For this second edition version, updated modifications/changes are supplemented in Subheadings 2, 3, and 4, and a new section describing the application of PCC in chromosome science fields is added with citation of updated references.

  12. Protein (Cyanobacteria): 186682931 [PGDBj - Ortholog DB

    Lifescience Database Archive (English)

    Full Text Available 63737:1993 ... hypothetical protein Npun_R2630 Nostoc punctiforme PCC 73102 MDTLDLQSVSTEDVMLRYGIKSRTTLNKFLENAGVNSFKEGRKTFIRMYQLGVLDRSAH...ELNYPINQSSNQSIQSIHPTDSIKSEQMELAESTGLFPLTTVDLLYITCEYENLPRLAKWLAGYAFLEKMSSGRVILPRDVVLKILDYKRLPTCKDGYFRYGNFVFLMIGDHKKEWLVSKK

  13. The epistemological and historical concept of nostoc from an indiciary lecture of George Canguilhem's "The cell theory"

    Directory of Open Access Journals (Sweden)

    Maurício de Carvalho Ramos

    2016-05-01

    Full Text Available Through an indiciary lecture of Canguilhem's article  "The cell theory", I propose, methodically guided by a epistemological-historical style research, the construction of the concept of nostoc as a metamorphic concept that integrated the concepts of primordial blastema and biblical slime of the French botanist Charles Naudin, set up a scientific culture of broad reach committed to solving the riddle of the morphological unity of organic and vital beings. The concept of Nostoc refers alchemically a gelatinous substance from the stars and endowed with balsamic medical virtues and botanically, a cianoficea algae of Nostoc genus. Examined the ideas of Canguilhem, Naudin, Jung and Paracelsus, this concept is proposed as a nucleoplasmatic oscillation, understood as a specific form of expression of the mythical-scientific theme of tension between continuity and discontinuity, as shown by Canguilhem in his history of cell concept.

  14. [Promotion effects of vitamin B12 on the degradation of 2, 4, 4'-trichlorobiphenyl by Nostoc PD-2].

    Science.gov (United States)

    Liu, Jia-Yu; Xiao, Wen-Feng; Lu, Li-Ping; Zhang, Hang-Jun

    2014-08-01

    Polychlorinated biphenyls are typical persistent chlorinated organic compounds in the environment. Bioremediation of PCB-contaminated environment has become one of the hot issues. In this study, vitamin B12 (VB12) and chlorine-free culture medium were applied to study the effects of VB12 on the degradation of 2,4,4'-trichlorobiphenyl (PCB28) by Nostoc PD-2 and the gene expression during the PCB-degradation process. Results showed that addition of different concentrations of vitamin B12 could improve the PCB-biodegradation rates by Nostoc PD-2. Compared with the control group, the 7-day degradation rate in 10 microg x L(-1), 100 microg x L(-1), and 1 000 microg x L(-1) VB12-treated groups increased by 11.0%, 19.7%, and 21.9% , respectively. The degradation half-time decreased from 5.53 days (treated with 10 microg x L(-1) VB12) to 3.08 days (treated with 100 microg x L(-1) VB12). The expression of cytochrome b6f complex iron-sulfur protein gene and dioxygenase gene showed significant correlation with PCB28-degradation by Nostoc PD-2. While the expression of iron-sulfur protein gene showed more significant correlation with PCB28-degradation. Results in this study indicated that adding VB12 could promote PCB28-degradation by Nostoc PD-2. Moreover, VB12 addition improved the PCB-degradation activity of Nostoc PD-2 at the gene level. The above conclusions could provide a new choice for developing efficient bioremediation technology for PCB-contaminated environment and a new insight into the PCB-biodegradation mechanism by Nostoc PD-2.

  15. Development and application of the PCC biodosimetry method in emergency preparedness

    Energy Technology Data Exchange (ETDEWEB)

    Stricklin, D. (Swedish Defence Research Agency, FOI (Sweden)); Lindholm, C. (Finnish Radiation and Nuclear Safety Authority, STUK (Finland)); Jaworska, A. (Norwegian Radiation Protection Authority, NRPA (Norway))

    2008-07-15

    A method for biological assessment of radiation dose for specific application in emergency preparedness was developed. Premature chromosome condensation (PCC) was investigated to provide a potentially faster means of analysis and the ability to assess higher doses than with the dicentric assay which is routinely applied in biodosimetry today. A review of existing methods was made, followed by experiments determine optimal assay conditions, and evaluations to determination of optimal conditions and the most appropriate endpoints for analyses. Twelve different experimental conditions were examined with four different evaluation approaches. Aspects during optimization such as practicality, speed, and reliability were considered. The conclusion from these studies was a PCC protocol utilizing okadaic acid for induction of PCC cells in stimulated lymphocytes but without the use of colcemid for metaphase arrest with the subsequent evaluation of ring chromosomes. Well-defined criteria were established for evaluation of PCC cells and ring chromosome aberrations. An inter-calibration was made by comparing assessment of ring chromosomes between all three laboratories. Agreement was made to count only rings with observable open spaces or large, obvious rings without open spaces. Finally a dose response curve for the PCC method was prepared and a comparison of the PCC method to the traditional dicentric assay in triage mode was made. The triage method requires a minimal number of evaluations so that categorization of high, medium and low doses may be made in an emergency situation where large numbers of people should be evaluated. The comparison of the PCC method with the dicentric assay triage method indicated that the PCC assay performed superior to the dicentric assay for evaluation of samples at higher doses, however, the dicentric assay appeared to provide more accurate dose assessment at lower doses. This project suggests a PCC assay method for biological dose estimates

  16. Fundamental Frequency Estimation of the Speech Signal Compressed by MP3 Algorithm Using PCC Interpolation

    Directory of Open Access Journals (Sweden)

    MILIVOJEVIC, Z. N.

    2010-02-01

    Full Text Available In this paper the fundamental frequency estimation results of the MP3 modeled speech signal are analyzed. The estimation of the fundamental frequency was performed by the Picking-Peaks algorithm with the implemented Parametric Cubic Convolution (PCC interpolation. The efficiency of PCC was tested for Catmull-Rom, Greville and Greville two-parametric kernel. Depending on MSE, a window that gives optimal results was chosen.

  17. Isoprenoid hydrocarbons produced by thermal alteration of Nostoc muscorum and Rhodopseudomonas spheroides

    Science.gov (United States)

    Philp, R. P.; Brown, S.; Calvin, M.

    1978-01-01

    The potential of algae and photosynthetic bacteria to serve as precursors of kerogen was studied to determine what factors affect the relative rates of formation of precursor hydrocarbons. Cells of Nostoc muscorum and Rhodopseudomonas spheroides were subjected to thermal alteration (by heating samples in glass tubes sealed under nitrogen) for two, four, and twelve weeks. Both unextracted and extracted cells in the absence and presence of montmorillonite were investigated, and the isoprenoid hydrocarbons produced in these experiments were determined. Phytane and five isomeric phytenes were the main hydrocarbons observed; their relative rates of formation in the different experimental conditions are described. No phytadienes, pristane, or pristenes were detected.

  18. Sintesa Precipitated Calcium Carbonate (PCC) dari Cangkang Kerang Darah (Anadara Granosa) dengan Variasi Ukuran Partikel dan Waktu Karbonasi

    OpenAIRE

    Rahmawati, Lucy; Amri, Amun; Zultiniar, Zultiniar; Yelmida, Yelmida

    2015-01-01

    Precipitated Calcium Carbonate (PCC) is a product of the processing of natural materials containing calcium carbonate resulting from the precipitation process with high purity. Bloodcockle shell can be used as a source of calcium for precipitated Calcium Carbonate. The purpose of this study to produce PCC of waste shells blood with carbonation method and determine the particle size of the PCC and the best carbonation time. Synthesis performed using carbonation method by adding nitric acid to ...

  19. Life cycle as a stable trait in the evaluation of diversity of Nostoc from biofilms in rivers.

    Science.gov (United States)

    Mateo, Pilar; Perona, Elvira; Berrendero, Esther; Leganés, Francisco; Martín, Marta; Golubić, Stjepko

    2011-05-01

    The diversity within the genus Nostoc is still controversial and more studies are needed to clarify its heterogeneity. Macroscopic species have been extensively studied and discussed; however, the microscopic forms of the genus, especially those from running waters, are poorly known and likely represented by many more species than currently described. Nostoc isolates from biofilms of two Spanish calcareous rivers were characterized comparing the morphology and life cycle in two culture media with different levels of nutrients and also comparing the 16S rRNA gene sequences. The results showed that trichome shape and cellular dimensions varied considerably depending on the culture media used, whereas the characteristics expressed in the course of the life cycle remained stable for each strain independent of the culture conditions. Molecular phylogenetic analysis confirmed the distinction between the studied strains established on morphological grounds. A balanced approach to the evaluation of diversity of Nostoc in the service of autecological studies requires both genotypic information and the evaluation of stable traits. The results of this study show that 16S rRNA gene sequence similarity serves as an important criterion for characterizing Nostoc strains and is consistent with stable attributes, such as the life cycle. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  20. Feathermoss and epiphytic Nostoc cooperate differently: expanding the spectrum of plant–cyanobacteria symbiosis

    Science.gov (United States)

    Warshan, Denis; Espinoza, Josh L; Stuart, Rhona K; Richter, R Alexander; Kim, Sea-Yong; Shapiro, Nicole; Woyke, Tanja; C Kyrpides, Nikos; Barry, Kerrie; Singan, Vasanth; Lindquist, Erika; Ansong, Charles; Purvine, Samuel O; M Brewer, Heather; Weyman, Philip D; Dupont, Christopher L; Rasmussen, Ulla

    2017-01-01

    Dinitrogen (N2)-fixation by cyanobacteria in symbiosis with feathermosses is the primary pathway of biological nitrogen (N) input into boreal forests. Despite its significance, little is known about the cyanobacterial gene repertoire and regulatory rewiring needed for the establishment and maintenance of the symbiosis. To determine gene acquisitions and regulatory changes allowing cyanobacteria to form and maintain this symbiosis, we compared genomically closely related symbiotic-competent and -incompetent Nostoc strains using a proteogenomics approach and an experimental set up allowing for controlled chemical and physical contact between partners. Thirty-two gene families were found only in the genomes of symbiotic strains, including some never before associated with cyanobacterial symbiosis. We identified conserved orthologs that were differentially expressed in symbiotic strains, including protein families involved in chemotaxis and motility, NO regulation, sulfate/phosphate transport, and glycosyl-modifying and oxidative stress-mediating exoenzymes. The physical moss–cyanobacteria epiphytic symbiosis is distinct from other cyanobacteria–plant symbioses, with Nostoc retaining motility, and lacking modulation of N2-fixation, photosynthesis, GS-GOGAT cycle and heterocyst formation. The results expand our knowledge base of plant–cyanobacterial symbioses, provide a model of information and material exchange in this ecologically significant symbiosis, and suggest new currencies, namely nitric oxide and aliphatic sulfonates, may be involved in establishing and maintaining the cyanobacteria–feathermoss symbiosis. PMID:28800136

  1. Sequestration of chromium by exopolysaccharides of Nostoc and Gloeocapsa from dilute aqueous solutions

    Energy Technology Data Exchange (ETDEWEB)

    Sharma, Mona [Department of Environmental Science and Engineering, Guru Jambheshwar University of Science and Technology, Hisar-125 001 (India); Kaushik, Anubha [Department of Environmental Science and Engineering, Guru Jambheshwar University of Science and Technology, Hisar-125 001 (India)], E-mail: aks_10@yahoo.com; Somvir,; Bala, Kiran; Kamra, Anjana [Department of Environmental Science and Engineering, Guru Jambheshwar University of Science and Technology, Hisar-125 001 (India)

    2008-09-15

    This article reports the chromium removal potential of exopolysaccharides (EPS) of two indigenously isolated cyanobacterial strains, Gloeocapsa calcarea and Nostoc punctiforme. The biosorption was studied by varying pH from 2 to 6 and initial chromium concentration from 5 to 20 mg/L to find out the optimized conditions for maximum chromium removal by EPS. Two equilibrium models, Langmuir and Freundlich, were used to explain these results. The Freundlich model was found to be better applicable to the experimental data as compared to Langmuir as inferred from high value of coefficient of determination whereas the optimal conditions were found to be same for the two (pH 2 and initial chromium concentration 20 mg/L). EPS production by the two strains was also studied which was found to be higher for Gloeocapsa. On the basis of experimental results and model parameters, it can be inferred that the EPS extracted from Nostoc has comparatively high biosorption capacity and can be utilized for the removal of chromium from dilute aqueous solution. Adsorption of chromium on EPS was further confirmed by surface morphology observed in scanning electron micrographs.

  2. Nostoc sphaeroides Kütz, a candidate producer par excellence for CELSS

    Science.gov (United States)

    Wang, Gaohong; Hao, Zongjie; Liu, Yongding

    A lot of aquatic organisms could be regarded as suitable candidates par excellence in the establishment of CELSS, since they are relatively easy and fast to grow and resistant to changes in environmental condition as well as providing nutritious, protein-and vitamin-rich foods for the crew, which can fulfill the main functions of CELSS, including supplying oxygen, water and food, removing carbon dioxide and making daily life waste reusable. Our labotory has developed mass culture of Nostoc sphaeroides Kütz, which is one of traditional healthy food in China and. The oxygen evolution rate of the cyanobacterium is about 150 molO2.mg-1.h-1, and it usually grows into colony with size between 2-20mm, which is easy to be harvested. It also can be cultured with high density, which show that the productivity of the cyanobacterium in limited volume is higher than other microalgae. We had measured the nutrient content of the cyanobacterium and developed some Chinese Dishes and Soups with Nostoc sphaeroides Kütz, which showed that it was a good food for crew. Using remote sensing technique, we also investigated its growth in Closed System under microgravity by SHENZHOU-2 spacecraft in January 2001. We plan to develop suitable bioreactor with the cyanobacterium for supplying oxygen and food to crew in future.

  3. Feathermoss and epiphytic Nostoc cooperate differently: expanding the spectrum of plant-cyanobacteria symbiosis.

    Science.gov (United States)

    Warshan, Denis; Espinoza, Josh L; Stuart, Rhona K; Richter, R Alexander; Kim, Sea-Yong; Shapiro, Nicole; Woyke, Tanja; C Kyrpides, Nikos; Barry, Kerrie; Singan, Vasanth; Lindquist, Erika; Ansong, Charles; Purvine, Samuel O; M Brewer, Heather; Weyman, Philip D; Dupont, Christopher L; Rasmussen, Ulla

    2017-12-01

    Dinitrogen (N 2 )-fixation by cyanobacteria in symbiosis with feathermosses is the primary pathway of biological nitrogen (N) input into boreal forests. Despite its significance, little is known about the cyanobacterial gene repertoire and regulatory rewiring needed for the establishment and maintenance of the symbiosis. To determine gene acquisitions and regulatory changes allowing cyanobacteria to form and maintain this symbiosis, we compared genomically closely related symbiotic-competent and -incompetent Nostoc strains using a proteogenomics approach and an experimental set up allowing for controlled chemical and physical contact between partners. Thirty-two gene families were found only in the genomes of symbiotic strains, including some never before associated with cyanobacterial symbiosis. We identified conserved orthologs that were differentially expressed in symbiotic strains, including protein families involved in chemotaxis and motility, NO regulation, sulfate/phosphate transport, and glycosyl-modifying and oxidative stress-mediating exoenzymes. The physical moss-cyanobacteria epiphytic symbiosis is distinct from other cyanobacteria-plant symbioses, with Nostoc retaining motility, and lacking modulation of N 2 -fixation, photosynthesis, GS-GOGAT cycle and heterocyst formation. The results expand our knowledge base of plant-cyanobacterial symbioses, provide a model of information and material exchange in this ecologically significant symbiosis, and suggest new currencies, namely nitric oxide and aliphatic sulfonates, may be involved in establishing and maintaining the cyanobacteria-feathermoss symbiosis.

  4. Sequestration of chromium by exopolysaccharides of Nostoc and Gloeocapsa from dilute aqueous solutions

    International Nuclear Information System (INIS)

    Sharma, Mona; Kaushik, Anubha; Somvir,; Bala, Kiran; Kamra, Anjana

    2008-01-01

    This article reports the chromium removal potential of exopolysaccharides (EPS) of two indigenously isolated cyanobacterial strains, Gloeocapsa calcarea and Nostoc punctiforme. The biosorption was studied by varying pH from 2 to 6 and initial chromium concentration from 5 to 20 mg/L to find out the optimized conditions for maximum chromium removal by EPS. Two equilibrium models, Langmuir and Freundlich, were used to explain these results. The Freundlich model was found to be better applicable to the experimental data as compared to Langmuir as inferred from high value of coefficient of determination whereas the optimal conditions were found to be same for the two (pH 2 and initial chromium concentration 20 mg/L). EPS production by the two strains was also studied which was found to be higher for Gloeocapsa. On the basis of experimental results and model parameters, it can be inferred that the EPS extracted from Nostoc has comparatively high biosorption capacity and can be utilized for the removal of chromium from dilute aqueous solution. Adsorption of chromium on EPS was further confirmed by surface morphology observed in scanning electron micrographs

  5. PCC-ring induction in human lymphocytes exposed to gamma and neutron irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Lamadrid Boada, Ana I; Garcia Lima, Omar [Centro de Proteccion e Higiene de las Radiaciones (CPHR), La Habana (Cuba); Delbos, Martine; Voisin, Philipe; Roy, Laurence [Institut de Radioprotection et de Surete Nucleaire, Fontenay-aux-Roses (France)

    2008-07-01

    Dose-effect curves for dose assessment in Gamma and neutron overexposures to high doses are presented in this paper for the first time in literature. The relationships were obtained by plotting the Premature Chromosome Condensation -rings (PCC-R) frequencies in PCC lymphocytes obtained by chemical induction with Calyculin A in vitro, with radiation doses between 5 to 25 Gy. For the elaboration of these curves 9 676 PCC cells in G1 G2 and M stages were analyzed. The results were fitted to a lineal quadratic model in Gamma irradiation and showed saturation starting from 20 Gy. For neutron irradiation the data was fitted to a lineal quadratic model up to 10 Gy, and then a markedly cell cycle arrest and saturation was observed. These curves are of particular interest for victims exposed to doses exceeding 5 Gy where it is always very difficult to estimate a dose using the conventional technique. (author)

  6. PCC-ring induction in human lymphocytes exposed to gamma and neutron irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Lamadrid B, A I; Garcia L, O [CPHR, Calle 20 No. 4113 e/41 y 47, Playa, La Habana 11300 (Cuba); Delbos, M; Voisin, P; Roy, L [Institut de Radioprotection et de Surete Nucleaire, BP 17, 92262 Fontenay-aux-Roses (France)

    2006-07-01

    Dose-effect curves for dose assessment in Gamma and neutron overexposures to high doses are presented in this paper for the first time in literature. The relationships were obtained by plotting the Premature Chromosome Condensation -rings (PCC-R) frequencies in PCC Iymphocytes obtained by chemical induction with Calyculin A in vitro, with radiation doses between 5 to 25 Gy. For the elaboration of these curves 9 676 PCC cells in Gl G2 and M stages were analyzed. The results were fitted to a lineal quadratic model in Gamma irradiation. For neutron irradiation the data was fitted to a lineal quadratic model up to 10 Gy and then a markedly cell cycle arrest and saturation was observed. These curves are of particular interest for victims exposed to doses exceeding 5 Gy where it is always very difficult to estimate a dose using the conventional technique. (Author)

  7. PCC-ring induction in human lymphocytes exposed to gamma and neutron irradiation

    International Nuclear Information System (INIS)

    Lamadrid B, A.I.; Garcia L, O.; Delbos, M.; Voisin, P.; Roy, L.

    2006-01-01

    Dose-effect curves for dose assessment in Gamma and neutron overexposures to high doses are presented in this paper for the first time in literature. The relationships were obtained by plotting the Premature Chromosome Condensation -rings (PCC-R) frequencies in PCC Iymphocytes obtained by chemical induction with Calyculin A in vitro, with radiation doses between 5 to 25 Gy. For the elaboration of these curves 9 676 PCC cells in Gl G2 and M stages were analyzed. The results were fitted to a lineal quadratic model in Gamma irradiation. For neutron irradiation the data was fitted to a lineal quadratic model up to 10 Gy and then a markedly cell cycle arrest and saturation was observed. These curves are of particular interest for victims exposed to doses exceeding 5 Gy where it is always very difficult to estimate a dose using the conventional technique. (Author)

  8. PCC-ring induction in human lymphocytes exposed to gamma and neutron irradiation

    International Nuclear Information System (INIS)

    Lamadrid Boada, Ana I.; Garcia Lima, Omar; Delbos, Martine; Voisin, Philipe; Roy, Laurence

    2008-01-01

    Dose-effect curves for dose assessment in Gamma and neutron overexposures to high doses are presented in this paper for the first time in literature. The relationships were obtained by plotting the Premature Chromosome Condensation -rings (PCC-R) frequencies in PCC lymphocytes obtained by chemical induction with Calyculin A in vitro, with radiation doses between 5 to 25 Gy. For the elaboration of these curves 9 676 PCC cells in G1 G2 and M stages were analyzed. The results were fitted to a lineal quadratic model in Gamma irradiation and showed saturation starting from 20 Gy. For neutron irradiation the data was fitted to a lineal quadratic model up to 10 Gy, and then a markedly cell cycle arrest and saturation was observed. These curves are of particular interest for victims exposed to doses exceeding 5 Gy where it is always very difficult to estimate a dose using the conventional technique. (author)

  9. Effect of Hydraulic Activity on Crystallization of Precipitated Calcium Carbonate (PCC) for Eco-Friendly Paper

    Science.gov (United States)

    Kim, Jung-Ah; Han, Gi-Chun; Lim, Mihee; You, Kwang-Suk; Ryu, Miyoung; Ahn, Ji-Whan; Fujita, Toyohisa; Kim, Hwan

    2009-01-01

    Wt% of aragonite, a CaCO3 polymorph, increased with higher hydraulic activity (°C) of limestone in precipitated calcium carbonate (PCC) from the lime-soda process (Ca(OH)2-NaOH-Na2CO3). Only calcite, the most stable polymorph, was crystallized at hydraulic activity under 10 °C, whereas aragonite also started to crystallize over 10 °C. The crystallization of PCC is more dependent on the hydraulic activity of limestone than CaO content, a factor commonly used to classify limestone ores according to quality. The results could be effectively applied to the determination of polymorphs in synthetic PCC for eco-friendly paper manufacture. PMID:20087470

  10. Effect of water curing duration on strength behaviour of portland composite cement (PCC) mortar

    Science.gov (United States)

    Caronge, M. A.; Tjaronge, M. W.; Hamada, H.; Irmawaty, R.

    2017-11-01

    Cement manufacturing of Indonesia has been introduced Portland Composite Cement (PCC) to minimize the rising production cost of cement which contains 80% clinker and 20% mineral admixture. A proper curing is very important when the cement contains mineral admixture materials. This paper reports the results of an experimental study conducted to evaluate the effect of water curing duration on strength behaviour of PCC mortar. Mortar specimens with water to cement ratio of (W/C) 0.5 were casted. Compressive strength, flexural strength and concrete resistance were tested at 7, 28 and 91 days cured water. The results indicated that water curing duration is essential to continue the pozzolanic reaction in mortar which contributes to the development of strength of mortar made with PCC.

  11. Evaluation of MIT-SCAN-T2 for thickness quality control for PCC and HMA pavements : research project capsule.

    Science.gov (United States)

    2013-04-01

    Thickness is currently a pay item for portland cement concrete (PCC) pavements : and a quality control item for both PCC and hot mix asphalt (HMA) pavements. : A change in pavement thickness of 0.5 in. can result in a reduction of multiple : years of...

  12. UV-induced developmental abnormalities in the filamentous blue-green alga Nostoc linckia

    International Nuclear Information System (INIS)

    Tiwari, D.N.

    1978-01-01

    Germinating spores of Nostoc linckia showed higher resistance against UV-irradiation compared to resting spores, maximal resistance being attained more rapidly in the presence of ammonium nitrogen. UV-irradiated germinating spores on further growth formed colonies consisting of abnormally large and spheroidal cells under non-photoreactivating conditions. The formation and fate of these abnormal cells was followed in detail in a mutant clone (M-5) raised from such a colony. Many of these cells formed spores which on return to growth-conducdive conditions germinated giving rise to different types of germlings from the abnormals which in certain cases proved lethal. The possibility of a transient polyenergidic and/or heterozygous state of these 'giant' cells has been discussed. (author)

  13. Study on activity measurement of Nostoc flagelliforme cells based on color identification

    Science.gov (United States)

    Wang, Yizhong; Su, Jianyu; Liu, Tiegen; Kong, Fanzhi; Jia, Shiru

    2008-12-01

    In order to measure the activities of Nostoc flagelliforme cells, a new method based on color identification was proposed in this paper. N. flagelliforme cells were colored with fluoreseein diaeetate. Then, an image of colored N. flagelliforme cells was taken, and changed from RGB model to HIS model. Its histogram of hue H was calculated, which was used as the input of a designed BP network. The output of the BP network was the description of measured activity of N. flagelliforme cells. After training, the activity of N. flagelliforme cells was identified by the BP network according to the histogram of H of their colored image. Experiments were conducted with satisfied results to show the feasibility and usefulness of activity measurement of N. flagelliforme cells based on color identification.

  14. Effects of light wavelengths on extracellular and capsular polysaccharide production by Nostoc flagelliforme.

    Science.gov (United States)

    Han, Pei-pei; Sun, Ying; Jia, Shi-ru; Zhong, Cheng; Tan, Zhi-lei

    2014-05-25

    The influences of different wavelengths of light (red 660nm, yellow 590nm, green 520nm, blue 460nm, purple 400nm) and white light on extracellular polysaccharide (EPS) and capsular polysaccharide (CPS) production by Nostoc flagelliforme in liquid culture were demonstrated in this study. The results showed that, compared with white light, red and blue lights significantly increased both EPS and CPS production while yellow light reduced their production; purple and green lights stimulated EPS production but inhibited CPS formation. Nine constituent monosaccharides and one uronic acid were detected in both EPS and CPS, and their ratios showed significant differences among treatment with different light wavelengths. However, the advanced structure of EPS and CPS from various light conditions did not present obvious difference through Fourier transform infrared spectroscopy and X-ray diffraction characterization. These findings establish a basis for development of high-yielding polysaccharide production process and understanding their regulation. Copyright © 2014 Elsevier Ltd. All rights reserved.

  15. System analysis of salt and osmotic stress induced proteins in Nostoc muscorum and Bradyrhizobium japonicum

    Directory of Open Access Journals (Sweden)

    Vipin Kaithwas

    2017-06-01

    Full Text Available In this study the proteome response of the two diazotrophic organism’s viz. Nostoc muscorum and Bradyrhizobium japonicum exposed to salt (NaCl and osmotic (sucrose stresses was compared. Out of the total over expressed proteins; we have selected only three over expressed proteins viz. GroEL chaperonin, nitrogenase Mo-Fe protein and argininosuccinate synthase for further analysis, and then we analyzed the amino acid frequencies of all the three over expressed proteins. That led to the conclusion that amino acids e.g. alanine, glycine and valine that were energetically cheaper to produce were showing higher frequencies. This study would help in tracing the phylogenetic relationship between protein families.

  16. Composition and occurrence of lipid droplets in the cyanobacterium Nostoc punctiforme.

    Science.gov (United States)

    Peramuna, Anantha; Summers, Michael L

    2014-12-01

    Inclusions of neutral lipids termed lipid droplets (LDs) located throughout the cell were identified in the cyanobacterium Nostoc punctiforme by staining with lipophylic fluorescent dyes. LDs increased in number upon entry into stationary phase and addition of exogenous fructose indicating a role for carbon storage, whereas high-light stress did not increase LD numbers. LD accumulation increased when nitrate was used as the nitrogen source during exponential growth as compared to added ammonia or nitrogen-fixing conditions. Analysis of isolated LDs revealed enrichment of triacylglycerol (TAG), α-tocopherol, and C17 alkanes. LD TAG from exponential phase growth contained mainly saturated C16 and C18 fatty acids, whereas stationary phase LD TAG had additional unsaturated fatty acids characteristic of whole cells. This is the first characterization of cyanobacterial LD composition and conditions leading to their production. Based upon their abnormally large size and atypical location, these structures represent a novel sub-organelle in cyanobacteria.

  17. Immunomodulatory Potential of the Polysaccharide-Rich Extract from Edible Cyanobacterium Nostoc commune

    Directory of Open Access Journals (Sweden)

    Hui-Fen Liao

    2015-11-01

    Full Text Available A dry sample of Nostoc commune from an organic farm in Pingtung city (Taiwan was used to prepare polysaccharide-rich (NCPS extract. The conditioned medium (CM from NCPS-treated human peripheral blood (PB-mononuclear cells (MNC effectively inhibited the growth of human leukemic U937 cells and triggered differentiation of U937 monoblast cells into monocytic/macrophagic lines. Cytokine levels in MNC-CMs showed upregulation of granulocyte/macrophage-colony stimulatory factor and IL-1β and downregulation of IL-6 and IL-17 upon treatment with NCPS. Moreover, murine macrophage RAW264.7 cells treated with NCPS exhibited the stimulatory effects of nitric oxide and superoxide secretion, indicating that NCPS might activate the immunity of macrophages. Collectively, the present study demonstrates that NCPS from N. commune could be potentially used for macrophage activation and consequently inhibited the leukemic cell growth and induced monocytic/macrophagic differentiation.

  18. Accumulation of radioactive cesium released from Fukushima Daiichi Nuclear Power Plant in terrestrial cyanobacteria Nostoc commune.

    Science.gov (United States)

    Sasaki, Hideaki; Shirato, Susumu; Tahara, Tomoya; Sato, Kenji; Takenaka, Hiroyuki

    2013-01-01

    The Fukushima Daiichi Nuclear Power Plant accident released large amounts of radioactive substances into the environment and contaminated the soil of Tohoku and Kanto districts in Japan. Removal of radioactive material from the environment is an urgent problem, and soil purification using plants is being considered. In this study, we investigated the ability of 12 seed plant species and a cyanobacterium to accumulate radioactive material. The plants did not accumulate radioactive material at high levels, but high accumulation was observed in the terrestrial cyanobacterium Nostoc commune. In Nihonmatsu City, Fukushima Prefecture, N. commune accumulated 415,000 Bq/kg dry weight (134)Cs and 607,000 Bq kg(-1) dry weight (137)Cs. The concentration of cesium in N. commune tended to be high in areas where soil radioactivity was high. A cultivation experiment confirmed that N. commune absorbed radioactive cesium from polluted soil. These data demonstrated that radiological absorption using N. commune might be suitable for decontaminating polluted soil.

  19. Effect of UV-B on enzymes of nitrogen metabolism in the cyanobacterium Nostoc calcicola

    International Nuclear Information System (INIS)

    Kumar, A.; Sinha, R.P.; Häder, D. P.

    1996-01-01

    The effects of ultraviolet-B (UV-B; 280–315 nm) irradiation on nitrogenase and nitrate reductase (NR) activity have been studied in the filamentous and heterocystous N 2 -fixing cyanobacterium Nostoc calcicola. Exposure of cultures to UV-B (5W/m 2 ) for as little as 30 min caused complete inactivation of nitrogenase activity whereas nitrate reductase activity was stimulated twofold in comparison to one exposed to fluorescent white light. GS activity was also inhibited by UV-B treatment, but there was no total loss of activity even after 4 h. NR activity showed a gradual stimulation up to 4 h and thereafter it became constant. Stimulation was also obtained in reductant deficient cultures (12 h incubation in the dark) suggesting independence of NR of PS-II under UV-B. NR activity was also unaffected in the presence of DCMU, a known inhibitor of PS-II. However, both O 2 evolution and 14 CO 2 uptake were completely abolished following 30 min of UV-B treatment. Addition of the protein synthesis inhibitor chloramphenicol (25 μg/mL) to cultures did not show any inhibitory effect on NR activity. SDS-PAGE analysis of UV-B treated cultures elicited gradual loss of protein bands with increasing duration of exposure. Our findings suggest that UV-B irradiance has differential effects on the enzymes of the nitrogen metabolism in the cyanobacterium Nostoc calcicola. Further studies are needed to reveal the exact mechanism involved in the stimulation of NR activity by UV-B. Whether UV-B has a direct effect on NO 2 − accumulation in the cells needs detailed investigation. (author)

  20. Isolation and characterization of the small subunit of the uptake hydrogenase from the cyanobacterium Nostoc punctiforme.

    Science.gov (United States)

    Raleiras, Patrícia; Kellers, Petra; Lindblad, Peter; Styring, Stenbjörn; Magnuson, Ann

    2013-06-21

    In nitrogen-fixing cyanobacteria, hydrogen evolution is associated with hydrogenases and nitrogenase, making these enzymes interesting targets for genetic engineering aimed at increased hydrogen production. Nostoc punctiforme ATCC 29133 is a filamentous cyanobacterium that expresses the uptake hydrogenase HupSL in heterocysts under nitrogen-fixing conditions. Little is known about the structural and biophysical properties of HupSL. The small subunit, HupS, has been postulated to contain three iron-sulfur clusters, but the details regarding their nature have been unclear due to unusual cluster binding motifs in the amino acid sequence. We now report the cloning and heterologous expression of Nostoc punctiforme HupS as a fusion protein, f-HupS. We have characterized the anaerobically purified protein by UV-visible and EPR spectroscopies. Our results show that f-HupS contains three iron-sulfur clusters. UV-visible absorption of f-HupS has bands ∼340 and 420 nm, typical for iron-sulfur clusters. The EPR spectrum of the oxidized f-HupS shows a narrow g = 2.023 resonance, characteristic of a low-spin (S = ½) [3Fe-4S] cluster. The reduced f-HupS presents complex EPR spectra with overlapping resonances centered on g = 1.94, g = 1.91, and g = 1.88, typical of low-spin (S = ½) [4Fe-4S] clusters. Analysis of the spectroscopic data allowed us to distinguish between two species attributable to two distinct [4Fe-4S] clusters, in addition to the [3Fe-4S] cluster. This indicates that f-HupS binds [4Fe-4S] clusters despite the presence of unusual coordinating amino acids. Furthermore, our expression and purification of what seems to be an intact HupS protein allows future studies on the significance of ligand nature on redox properties of the iron-sulfur clusters of HupS.

  1. Genetic analysis reveals the identity of the photoreceptor for phototaxis in hormogonium filaments of Nostoc punctiforme.

    Science.gov (United States)

    Campbell, Elsie L; Hagen, Kari D; Chen, Rui; Risser, Douglas D; Ferreira, Daniela P; Meeks, John C

    2015-02-15

    In cyanobacterial Nostoc species, substratum-dependent gliding motility is confined to specialized nongrowing filaments called hormogonia, which differentiate from vegetative filaments as part of a conditional life cycle and function as dispersal units. Here we confirm that Nostoc punctiforme hormogonia are positively phototactic to white light over a wide range of intensities. N. punctiforme contains two gene clusters (clusters 2 and 2i), each of which encodes modular cyanobacteriochrome-methyl-accepting chemotaxis proteins (MCPs) and other proteins that putatively constitute a basic chemotaxis-like signal transduction complex. Transcriptional analysis established that all genes in clusters 2 and 2i, plus two additional clusters (clusters 1 and 3) with genes encoding MCPs lacking cyanobacteriochrome sensory domains, are upregulated during the differentiation of hormogonia. Mutational analysis determined that only genes in cluster 2i are essential for positive phototaxis in N. punctiforme hormogonia; here these genes are designated ptx (for phototaxis) genes. The cluster is unusual in containing complete or partial duplicates of genes encoding proteins homologous to the well-described chemotaxis elements CheY, CheW, MCP, and CheA. The cyanobacteriochrome-MCP gene (ptxD) lacks transmembrane domains and has 7 potential binding sites for bilins. The transcriptional start site of the ptx genes does not resemble a sigma 70 consensus recognition sequence; moreover, it is upstream of two genes encoding gas vesicle proteins (gvpA and gvpC), which also are expressed only in the hormogonium filaments of N. punctiforme. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  2. The mechanisms of protection of antioxidants on Nostoc sphaeroides against UV-B radiation

    Science.gov (United States)

    Wang, G. H.

    UV radiation is one of space harmful factor for earth organisms in space exploration In the present work we studied on the role of antioxidant system in Nostoc sphaeroides K u tz Cyanobacteria and the effects of exogenous antioxidant molecules on its photosynthetic rate under UV-B radiation It was found that UV-B radiation decreased the photosynthetic activity of cyanobacterium but promoted the activity of antioxidant system to protect photosystem II PSII and exogenous antioxidant sodium nitroprusside SNP N-acetylcysteine NAC had an obvious protection on PSII activity under UV-B radiation The activity of SOD Superoxide Dismutase EC 1 15 1 1 CAT Catalase EC 1 11 1 6 POD Peroxidase EC 1 11 1 7 and content of MDA and ASC were improved by 0 5mM and 1mM SNP but 0 1mM SNP decreased the activity of antioxide system Exogenous NAC addition decreased the activity of SOD POD CAT and the content MDA and ASC but exogenous NAC addition increased the content of GSH The results suggested that exogenous SNP and NAC may protect algae by different mechanisms in which SNP maybe play double roles as sources of reactive free radicals or ROS scavengers in formation of algae s protection of PSII under UV-B radiation while NAC does function as antioxidant reagent or precursor of glutathione which could protect PSII directly from UV-B radiation Keyword antioxidant system exogenous or endogenous antioxidant Nostoc sphaeroides photosynthesis UV-B radiation

  3. Isolation and in silico analysis of Fe-superoxide dismutase in the cyanobacterium Nostoc commune.

    Science.gov (United States)

    Kesheri, Minu; Kanchan, Swarna; Richa; Sinha, Rajeshwar P

    2014-12-15

    Cyanobacteria are known to endure various stress conditions due to the inbuilt potential for oxidative stress alleviation owing to the presence of an array of antioxidants. The present study shows that Antarctic cyanobacterium Nostoc commune possesses two antioxidative enzymes viz., superoxide dismutase (SOD) and catalase that jointly cope with environmental stresses prevailing at its natural habitat. Native-PAGE analysis illustrates the presence of a single prominent isoform recognized as Fe-SOD and three distinct isoforms of catalase. The protein sequence of Fe-SOD in N. commune retrieved from NCBI protein sequence database was used for in silico analysis. 3D structure of N. commune was predicted by comparative modeling using MODELLER 9v11. Further, this model was validated for its quality by Ramachandran plot, ERRAT, Verify 3D and ProSA-web which revealed good structure quality of the model. Multiple sequence alignment showed high conservation in N and C-terminal domain regions along with all metal binding positions in Fe-SOD which were also found to be highly conserved in all 28 cyanobacterial species under study, including N. commune. In silico prediction of isoelectric point and molecular weight of Fe-SOD was found to be 5.48 and 22,342.98Da respectively. The phylogenetic tree revealed that among 28 cyanobacterial species, Fe-SOD in N. commune was the closest evolutionary homolog of Fe-SOD in Nostoc punctiforme as evident by strong bootstrap value. Thus, N. commune may serve as a good biological model for studies related to survival of life under extreme conditions prevailing at the Antarctic region. Moreover cyanobacteria may be exploited for biochemical and biotechnological applications of enzymatic antioxidants. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Separation of Aeruginosin-865 from Cultivated Soil Cyanobacterium (Nostoc sp.) by Centrifugal Partition Chromatography combined with Gel Permeation Chromatography.

    Science.gov (United States)

    Cheel, José; Minceva, Mirjana; Urajová, Petra; Aslam, Rabya; Hrouzek, Pavel; Kopecký, Jiří

    2015-10-01

    Aeruginosin-865 was isolated from cultivated soil cyanobacteria using a combination of centrifugal partition chromatography (CPC) and gel permeation chromatography. The solubility of Aer-865 in different solvents was evaluated using the conductor-like screening model for real solvents (COSMO-RS). The CPC separation was performed in descending mode with a biphasic solvent system composed of water-n-BuOH-acetic acid (5:4:1, v/v/v). The upper phase was used as a stationary phase, whereas the lower phase was employed as a mobile phase at a flow rate of 10 mL/min. The revolution speed and temperature of the separation column were 1700 rpm and 25 degrees C, respectively. Preparative CPC separation followed by gel permeation chromatography was performed on 50 mg of crude extract yielding Aer-865 (3.5 mg), with a purity over 95% as determined by HPLC. The chemical identity of the isolated compound was confirmed by comparing its spectroscopic data (UV, HRESI-MS, HRESI-MS/MS) with those of an authentic standard and data available in the literature.

  5. Two-Step Separation of Nostotrebin 6 from Cultivated Soil Cyanobacterium (Nostoc sp. by High Performance Countercurrent Chromatography

    Directory of Open Access Journals (Sweden)

    José Cheel

    2014-06-01

    Full Text Available High performance countercurrent chromatography (HPCCC was successfully applied for the separation of nostotrebin 6 from cultivated soil cyanobacteria in a two-step operation. A two-phase solvent system composed of n-hexane–ethyl acetate–methanol–water (4:5:4:5, v/v/v/v was employed for the HPCCC separation. In the first-step operation, its neutral upper phase was used as stationary phase and its basic lower phase (1% NH3 in lower phase was employed as mobile phase at a flow rate of 1 mL/min. In the second operation step, its neutral upper phase was used as stationary phase, whereas both its neutral lower phase and basic lower phase were employed as mobile phase with a linear gradient elution at a flow rate of 0.8 mL/min. The revolution speed and temperature of the separation column were 1,000 rpm and 30 °C, respectively. Using HPCCC followed by clean-up on Sephadex LH-20 gel, 4 mg of nostotrebin 6 with a purity of 99% as determined by HPLC/DAD-ESI-HRMS was obtained from 100 mg of crude extract. The chemical identity of the isolated compound was confirmed by comparing its spectroscopic data (UV, ESI-HRMS, ESI-HRMS2 with those of an authentic standard and data available in the literature.

  6. Separation of Aeruginosin-865 from Cultivated Soil Cyanobacterium (Nostoc sp.) by Centrifugal Partition Chromatography combined with Gel Permeation Chromatography

    Czech Academy of Sciences Publication Activity Database

    Cheel, José; Minceva, M.; Urajová, Petra; Aslam, R.; Hrouzek, Pavel; Kopecký, Jiří

    2015-01-01

    Roč. 10, č. 10 (2015), s. 1719-1722 ISSN 1934-578X R&D Projects: GA MŠk EE2.3.30.0059; GA MŠk ED2.1.00/03.0110; GA MŠk LO1416 Institutional support: RVO:61388971 Keywords : Cyanobacteria * Aeruginosin-865 * Centrifugal partition chromatography Subject RIV: EE - Microbiology, Virology Impact factor: 0.884, year: 2015

  7. Dose - Response Curves for Dicentrics and PCC Rings: Preparedness for Radiological Emergency in Thailand

    International Nuclear Information System (INIS)

    Rungsimaphorn, B.; Rerkamnuaychoke, B.; Sudprasert, W.

    2014-01-01

    Establishing in-vitro dose calibration curves is important for reconstruction of radiation dose in the exposed individuals. The aim of this pioneering work in Thailand was to generate dose-response curves using conventional biological dosimetry: dicentric chromosome assay (DCA) and premature chromosome condensation (PCC) assay. The peripheral blood lymphocytes were irradiated with 137 Cs at a dose rate of 0.652 Gy/min to doses of 0.1, 0.25, 0.5, 0.75, 1, 2, 3, 4 and 5 Gy for DCA technique, and 5, 10, 15, 20 and 25 Gy for PCC technique. The blood samples were cultured and processed following the standard procedure given by the IAEA with slight modifications. At least 500-1,000 metaphases or 100 dicentrics/ PCC rings were analyzed using an automated metaphase finder system. The yield of dicentrics with dose was fitted to a linear quadratic model using Chromosome Aberration Calculation Software (CABAS, version 2.0), whereas the dose-response curve of PCC rings was fitted to a linear relationship. These curves will be useful for in-vitro dose reconstruction and can support the preparedness for radiological emergency in the country.

  8. LOFT CIS analysis: 1''-PCC-76-A inside containment penetration S-1A. Internal technical report

    International Nuclear Information System (INIS)

    Nitzel, M.E.

    1978-01-01

    The stress analysis performed on the 1''-PCC-76-A piping system inside containment penetration S-1A is described. Deadweight, thermal expansion, and seismic loads were considered. Results of this analysis show that the subject piping system will meet ASME Boiler and Pressure Vessel Code, Section III, Class 2 requirements provided that supports S8 and S9 are installed as recommended

  9. Factors Associated with Parental Satisfaction with a Pediatric Crisis Clinic (PCC).

    Science.gov (United States)

    Lee, Jonathan; Korczak, Daphne

    2014-05-01

    Little is known about parental satisfaction with pediatric crisis clinics (PCCs) that provide a single consultation to families in need of urgent psychiatric care. Parental satisfaction may improve long-term adherence to physician recommendations. To explore parental satisfaction with a PCC. Parental satisfaction was ascertained by a structured telephone interview following crisis consultation at the PCC of an academic, tertiary care centre. Parents of 71% (n = 124) of 174 pediatric patients seen in the PCC from 2007-2008 participated in the post-consultation interview. The majority of parents stated they were either somewhat satisfied (49/122, 40.2%) or very satisfied (49/122, 40.2%) with the PCC. Parental satisfaction correlated with time between referral and consultation (p<0.05), the degree to which parents felt listened to by the consultant (p<0.01), the amount of psychoeducation parents felt they received (p<0.01), and appointment length (p<0.001). Parents were satisfied overall with an urgent care service model. Satisfaction was correlated with the time between referral and consultation, degree to which they felt their consultant had listened to them, and the amount of information they received at the consultation's conclusion.

  10. Identification of the n-1 fatty acid as an antibacterial constituent from the edible freshwater cyanobacterium Nostoc verrucosum.

    Science.gov (United States)

    Oku, Naoya; Yonejima, Kohsuke; Sugawa, Takao; Igarashi, Yasuhiro

    2014-01-01

    The cyanobacterium Nostoc verrucosum occurs in cool, clear streams and its gelatinous colonies, called "ashitsuki," have been eaten in ancient Japan. Its ethanolic extract was found to inhibit the growth of Gram-positive bacteria and activity-guided fractionation yielded an unusual n-1 fatty acid, (9Z,12Z)-9,12,15-hexadecatrienoic acid (1), as one of the active principles. It inhibited the growth of Staphylococcus aureus at MIC 64 μg/mL.

  11. The global response of Nostoc punctiforme ATCC 29133 to UVA stress, assessed in a temporal DNA microarray study.

    Science.gov (United States)

    Soule, Tanya; Gao, Qunjie; Stout, Valerie; Garcia-Pichel, Ferran

    2013-01-01

    Cyanobacteria in nature are exposed not only to the visible spectrum of sunlight but also to its harmful ultraviolet components (UVA and UVB). We used Nostoc punctiforme ATCC 29133 as a model to study the UVA response by analyzing global gene expression patterns using genomic microarrays. UVA exposure resulted in the statistically detectable differential expression of 573 genes of the 6903 that were probed, compared with that of the control cultures. Of those genes, 473 were up-regulated, while only 100 were down-regulated. Many of the down-regulated genes were involved in photosynthetic pigment biosynthesis, indicating a significant shift in this metabolism. As expected, we detected the up-regulation of genes encoding antioxidant enzymes and the sunscreen, scytonemin. However, a majority of the up-regulated genes, 47%, were unassignable bioinformatically to known functional categories, suggesting that the UVA stress response is not well understood. Interestingly, the most dramatic up-regulation involved several contiguous genes of unassigned metabolism on plasmid A. This is the first global UVA stress response analysis of any phototrophic microorganism and the differential expression of 8% of the genes of the Nostoc genome indicates that adaptation to UVA in Nostoc has been an evolutionary force of significance. © 2012 Wiley Periodicals, Inc. Photochemistry and Photobiology © 2012 The American Society of Photobiology.

  12. Reconstruction of structural evolution in the trnL intron P6b loop of symbiotic Nostoc (Cyanobacteria).

    Science.gov (United States)

    Olsson, Sanna; Kaasalainen, Ulla; Rikkinen, Jouko

    2012-02-01

    In this study we reconstruct the structural evolution of the hyper-variable P6b region of the group I trnLeu intron in a monophyletic group of lichen-symbiotic Nostoc strains and establish it as a useful marker in the phylogenetic analysis of these organisms. The studied cyanobacteria occur as photosynthetic and/or nitrogen-fixing symbionts in lichen species of the diverse Nephroma guild. Phylogenetic analyses and secondary structure reconstructions are used to improve the understanding of the replication mechanisms in the P6b stem-loop and to explain the observed distribution patterns of indels. The variants of the P6b region in the Nostoc clade studied consist of different combinations of five sequence modules. The distribution of indels together with the ancestral character reconstruction performed enables the interpretation of the evolution of each sequence module. Our results indicate that the indel events are usually associated with single nucleotide changes in the P6b region and have occurred several times independently. In spite of their homoplasy, they provide phylogenetic information for closely related taxa. Thus we recognize that features of the P6b region can be used as molecular markers for species identification and phylogenetic studies involving symbiotic Nostoc cyanobacteria.

  13. Expression of organophosphorus-degradation gene ( opd) in aggregating and non-aggregating filamentous nitrogen-fixing cyanobacteria

    Science.gov (United States)

    Li, Qiong; Tang, Qing; Xu, Xudong; Gao, Hong

    2010-11-01

    Genetic engineering in filamentous N2-fixing cyanobacteria usually involves Anabaena sp. PCC 7120 and several other non-aggregating species. Mass culture and harvest of such species are more energy consuming relative to aggregating species. To establish a gene transfer system for aggregating species, we tested many species of Anabaena and Nostoc, and identified Nostoc muscorum FACHB244 as a species that can be genetically manipulated using the conjugative gene transfer system. To promote biodegradation of organophosphorus pollutants in aquatic environments, we introduced a plasmid containing the organophosphorus-degradation gene ( opd) into Anabaena sp. PCC 7120 and Nostoc muscorum FACHB244 by conjugation. The opd gene was driven by a strong promoter, P psbA . From both species, we obtained transgenic strains having organophosphorus-degradation activities. At 25°C, the whole-cell activities of the transgenic Anabaena and Nostoc strains were 0.163±0.001 and 0.289±0.042 unit/μg Chl a, respectively. However, most colonies resulting from the gene transfer showed no activity. PCR and DNA sequencing revealed deletions or rearrangements in the plasmid in some of the colonies. Expression of the green fluorescent protein gene from the same promoter in Anabaena sp. PCC 7120 showed similar results. These results suggest that there is the potential to promote the degradation of organophosphorus pollutants with transgenic cyanobacteria and that selection of high-expression transgenic colonies is important for genetic engineering of Anabaena and Nostoc species. For the first time, we established a gene transfer and expression system in an aggregating filamentous N2-fixing cyanobacterium. The genetic manipulation system of Nostoc muscorum FACHB244 could be utilized in the elimination of pollutants and large-scale production of valuable proteins or metabolites.

  14. Highly plastic genome of Microcystis aeruginosa PCC 7806, a ubiquitous toxic freshwater cyanobacterium

    Directory of Open Access Journals (Sweden)

    Latifi Amel

    2008-06-01

    Full Text Available Abstract Background The colonial cyanobacterium Microcystis proliferates in a wide range of freshwater ecosystems and is exposed to changing environmental factors during its life cycle. Microcystis blooms are often toxic, potentially fatal to animals and humans, and may cause environmental problems. There has been little investigation of the genomics of these cyanobacteria. Results Deciphering the 5,172,804 bp sequence of Microcystis aeruginosa PCC 7806 has revealed the high plasticity of its genome: 11.7% DNA repeats containing more than 1,000 bases, 6.8% putative transposases and 21 putative restriction enzymes. Compared to the genomes of other cyanobacterial lineages, strain PCC 7806 contains a large number of atypical genes that may have been acquired by lateral transfers. Metabolic pathways, such as fermentation and a methionine salvage pathway, have been identified, as have genes for programmed cell death that may be related to the rapid disappearance of Microcystis blooms in nature. Analysis of the PCC 7806 genome also reveals striking novel biosynthetic features that might help to elucidate the ecological impact of secondary metabolites and lead to the discovery of novel metabolites for new biotechnological applications. M. aeruginosa and other large cyanobacterial genomes exhibit a rapid loss of synteny in contrast to other microbial genomes. Conclusion Microcystis aeruginosa PCC 7806 appears to have adopted an evolutionary strategy relying on unusual genome plasticity to adapt to eutrophic freshwater ecosystems, a property shared by another strain of M. aeruginosa (NIES-843. Comparisons of the genomes of PCC 7806 and other cyanobacterial strains indicate that a similar strategy may have also been used by the marine strain Crocosphaera watsonii WH8501 to adapt to other ecological niches, such as oligotrophic open oceans.

  15. TetR Family Transcriptional Regulator PccD Negatively Controls Propionyl Coenzyme A Assimilation in Saccharopolyspora erythraea.

    Science.gov (United States)

    Xu, Zhen; Wang, Miaomiao; Ye, Bang-Ce

    2017-10-15

    Propanol stimulates erythromycin biosynthesis by increasing the supply of propionyl coenzyme A (propionyl-CoA), a starter unit of erythromycin production in Saccharopolyspora erythraea Propionyl-CoA is assimilated via propionyl-CoA carboxylase to methylmalonyl-CoA, an extender unit of erythromycin. We found that the addition of n -propanol or propionate caused a 4- to 16-fold increase in the transcriptional levels of the SACE_3398-3400 locus encoding propionyl-CoA carboxylase, a key enzyme in propionate metabolism. The regulator PccD was proved to be directly involved in the transcription regulation of the SACE_3398-3400 locus by EMSA and DNase I footprint analysis. The transcriptional levels of SACE_3398-3400 were upregulated 15- to 37-fold in the pccD gene deletion strain (Δ pccD ) and downregulated 3-fold in the pccD overexpression strain (WT/pIB- pccD ), indicating that PccD was a negative transcriptional regulator of SACE_3398-3400. The Δ pccD strain has a higher growth rate than that of the wild-type strain (WT) on Evans medium with propionate as the sole carbon source, whereas the growth of the WT/pIB- pccD strain was repressed. As a possible metabolite of propionate metabolism, methylmalonic acid was identified as an effector molecule of PccD and repressed its regulatory activity. A higher level of erythromycin in the Δ pccD strain was observed compared with that in the wild-type strain. Our study reveals a regulatory mechanism in propionate metabolism and suggests new possibilities for designing metabolic engineering to increase erythromycin yield. IMPORTANCE Our work has identified the novel regulator PccD that controls the expression of the gene for propionyl-CoA carboxylase, a key enzyme in propionyl-CoA assimilation in S. erythraea PccD represses the generation of methylmalonyl-CoA through carboxylation of propionyl-CoA and reveals an effect on biosynthesis of erythromycin. This finding provides novel insight into propionyl-CoA assimilation, and

  16. Protein (Cyanobacteria): 218248342 [PGDBj - Ortholog DB

    Lifescience Database Archive (English)

    Full Text Available ... hypothetical protein PCC8801_3595 Cyanothece sp. PCC 8801 MSIQYLLDENLPHLYREQLLRLKSDLTVWIIGDPGVPPKSTLDPEILIWCEQNKFILVTNNRASMPVHLADHLSQNRHIPGIFVLRPKASIGEIIDDLILIDELGNPQDYQDCISHIPFI

  17. SP. Pescado

    Directory of Open Access Journals (Sweden)

    Renato Gendre

    2003-12-01

    Full Text Available Nell'occhiello di un articolo dal titolo Il Peru dei de[Jini rosa e de/la grande pioggia si legge: "da una partenza  in aereo al «pescado»  che ti  sfamera."1 Questa parola spagnola, giustamente chiusa tra caporali, a noi pare molto interes­ sante, perche, nonostante l'apparenza, non ha nulla da spartire sotto i1 profilo se­ mantico con l'it. pescato. lnfatti, tutti i piu importanti dizionari della lingua italiana, di ieri e di oggi, etimologici e non 2, registrano  accanto a pescata,  ii lemma pescato, 3 ma lo spiegano come "quantita di pesce catturato nel corso di una battuta o di una stagione di pesca",4 mentre lo sp. pescado  indica i1 "pesce (solo nel senso di: pesGe pescato da mangiare [...]".s

  18. United States Air Force Research on Airfield Pavement Repairs Using Precast Portland Cement Concrete (PCC) Slabs (BRIEFING SLIDES)

    National Research Council Canada - National Science Library

    Saeed, Athar

    2008-01-01

    ...) slab repairs using precast PCC slab panels. AFRL is leading the technology development by critically reviewing the research conducted to date in this arena by the Air Force and the highway and civil aviation agencies and adopting...

  19. Validation Data Acquisition in HTTF during PCC Events

    Energy Technology Data Exchange (ETDEWEB)

    Bardet, Philippe [George Washington Univ., Washington, DC (United States)

    2018-02-07

    A validation experimental campaign was conducted in an Integral Effect Test (IET) facility of High Temperature Gas Reactors (HTGR), the High-Temperature Test Facility (HTTF) at Oregon State University (OSU). The HTTF simulates Depressurized and Pressurized Conduction Cooldowns (DCC and PCC). This campaign required the development of a new laser spectroscopic diagnostic to measure velocity in the challenging conditions of the HTTF: low speed (~1 m/s) gas flows at HTGR prototypical temperature and 1/10th pressure. This was a collaborative effort between co-PIs at The George Washington University (GW), Oregon State University (OSU), and NASA Langley Research Center. The main accomplishments of this project include the record for dynamic range for velocimetry, highest accuracy obtained with this technique, successful deployment in an IET leading to new validation matrix for CFD. These are detailed below and in manuscript appended to this executive summary. For this project, we introduced a new class of laser spectroscopic diagnostics to Thermal-Hydraulics to measure velocity of gases; furthermore, the diagnostic was demonstrated in-situ in an IET during DCC events. In such scenarios, particles used in mainstream techniques, like Particle Image Velocimetry (PIV) are not appropriate as they settle down too rapidly and also contaminate the experimental facility. Molecular tracers stay mixed within the working gas and can seed the flow in a technique called Molecular Tagging Velocimetry (MTV). In MTV a molecular tracer is photo-dissociated by a first (write) laser into radicals or molecules. The pattern created by the write laser is then interrogated with planar laser-induced fluorescence (PLIF), the read pulse(s), which are recorded with a camera. The pattern is probed and matched at two times (interval or probe time, dt), resulting in a time-of-flight velocimetry technique. This project demonstrated a new application range for MTV in gases. MTV has been extensively used

  20. Nezha, a novel active miniature inverted-repeat transposable element in cyanobacteria

    International Nuclear Information System (INIS)

    Zhou Fengfeng; Tran Thao; Xu Ying

    2008-01-01

    Miniature inverted-repeat transposable elements (MITEs) were first identified in plants and exerted extensive proliferations throughout eukaryotic and archaeal genomes. But very few MITEs have been characterized in bacteria. We identified a novel MITE, called Nezha, in cyanobacteria Anabaena variabilis ATCC 29413 and Nostoc sp. PCC 7120. Nezha, like most previously known MITEs in other organisms, is small in size, non-coding, carrying TIR and DR signals, and of potential to form a stable RNA secondary structure, and it tends to insert into A+T-rich regions. Recent transpositions of Nezha were observed in A. variabilis ATCC 29413 and Nostoc sp. PCC 7120, respectively. Nezha might have proliferated recently with aid from the transposase encoded by ISNpu3-like elements. A possible horizontal transfer event of Nezha from cyanobacteria to Polaromonas JS666 is also observed

  1. Blueprint for a minimal photoautotrophic cell: conserved and variable genes in Synechococcus elongatus PCC 7942

    Directory of Open Access Journals (Sweden)

    Peretó Juli

    2011-01-01

    Full Text Available Abstract Background Simpler biological systems should be easier to understand and to engineer towards pre-defined goals. One way to achieve biological simplicity is through genome minimization. Here we looked for genomic islands in the fresh water cyanobacteria Synechococcus elongatus PCC 7942 (genome size 2.7 Mb that could be used as targets for deletion. We also looked for conserved genes that might be essential for cell survival. Results By using a combination of methods we identified 170 xenologs, 136 ORFans and 1401 core genes in the genome of S. elongatus PCC 7942. These represent 6.5%, 5.2% and 53.6% of the annotated genes respectively. We considered that genes in genomic islands could be found if they showed a combination of: a unusual G+C content; b unusual phylogenetic similarity; and/or c a small number of the highly iterated palindrome 1 (HIP1 motif plus an unusual codon usage. The origin of the largest genomic island by horizontal gene transfer (HGT could be corroborated by lack of coverage among metagenomic sequences from a fresh water microbialite. Evidence is also presented that xenologous genes tend to cluster in operons. Interestingly, most genes coding for proteins with a diguanylate cyclase domain are predicted to be xenologs, suggesting a role for horizontal gene transfer in the evolution of Synechococcus sensory systems. Conclusions Our estimates of genomic islands in PCC 7942 are larger than those predicted by other published methods like SIGI-HMM. Our results set a guide to non-essential genes in S. elongatus PCC 7942 indicating a path towards the engineering of a model photoautotrophic bacterial cell.

  2. Safety study of PCC 2140 and ALILOG 21 used as part of safety measurement systems

    International Nuclear Information System (INIS)

    Meriaux, Pierre; Adnot, Serge; Rayrolles, Catherine.

    1978-03-01

    The PCC 2140 and ALILOG 21 equipment may be used at C.E.A. or E.D.F., as part of safety measurement systems. In a study of a similar, but earlier equipment, it was noticed that certain types of failures caused the system to switch to the least sensitive measurement range, which was detrimental to safety. This report analyses failure modes leading to unsafe failures and evaluates the risks ran into taking in account tests during use [fr

  3. Antioxidative activity of ethanol extracts from Spirulina platensis and Nostoc linckia measured by various methods

    Directory of Open Access Journals (Sweden)

    Liliana CEPOI

    2009-11-01

    Full Text Available The goal of this work is to determine the level of antioxidative activity of various ethanol extracts from Spirulina platensis and Nostoc linckia biomass, and also to demonstrate the possibility to select suitable methods for evaluation of these characteristics. The methods for determination of antioxidative activity were selected concerning their possible use for complex preparations: phosphomolybdenum method for evaluation of antioxidant capacity (PMRC, radical-scavenging activity by DPPH method (DPPH, antioxidant activity by the ABTS+ radical cation assay (ABTS, Folin-Ciocalteu reducing capacity (FCRC. We showed the presence of antioxidative substances in ethanol extractions from 2 species of cyanobacteria, and possibility to increase their activity varying ethanol concentration. It facilitates the extraction both water- and lipid-soluble components from biomass. Regarding used methods for antioxidative activity determination, we have used only those based on reaction of electrons return (which widely used nowadays in vitro. Obtained in different ways results demonstrate high reduction capacity of the extracts and possibility to select suitable analytical methods for each case.

  4. Investigation on characteristics of 3D printing using Nostoc sphaeroides biomass.

    Science.gov (United States)

    An, Yan-Jun; Guo, Chao-Fan; Zhang, Min; Zhong, Ze-Ping

    2018-06-27

    Gel-like constructs can be produced using an extrusion-based 3D food printing (3D-FP) technique. Nostoc sphaeroides biomass is a natural gel material. Considering its good nutrition and rheological properties, these algae were chosen in this study as supply material (ink) for 3D-FP. With this gel material, the extrusion-based 3D printing system was set as a model, and the printing behavior was investigated. Furthermore, the 3D-FP mechanisms were explained through low-field nuclear magnetic resonance and rheological measurements. Results indicated that although fresh biomass gel was printable, nonuniformity and instability occurred during printing. Blanched inks showed nonsmooth printing behavior, which was associated with a decrease in elasticity and viscosity. The printability was improved by increasing the rehydration time to 24 h when rehydrated powder was used. Increasing the rehydration time increased the water-binding degree. Pre-gelatinized potato starch was added to the mixture at ratios ranging from 1 to 100 g kg -1 . The best printing outcome was observed at 40 g kg -1 potato starch. We emphasize that elasticity and viscosity balance is an essential parameter to achieve printability. The strategies adopted in this work provide new insights into the development of personalized food regarding texture and nutritional additive content. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  5. Provision of water by halite deliquescence for Nostoc commune biofilms under Mars relevant surface conditions

    Science.gov (United States)

    Jänchen, Jochen; Feyh, Nina; Szewzyk, Ulrich; de Vera, Jean-Pierre P.

    2016-04-01

    Motivated by findings of new mineral related water sources for organisms under extremely dry conditions on Earth we studied in an interdisciplinary approach the water sorption behaviour of halite, soil component and terrestrial Nostoc commune biofilm under Mars relevant environmental conditions. Physicochemical methods served for the determination of water sorption equilibrium data and survival of heterotrophic bacteria in biofilm samples with different water contents was assured by recultivation. Deliquescence of halite provides liquid water at temperatures <273 K and may serve as water source on Mars during the morning stabilized by the CO2 atmosphere for a few hours. The protecting biofilm of N. commune is rather hygroscopic and tends to store water at lower humidity values. Survival tests showed that a large proportion of the Alphaproteobacteria dominated microbiota associated to N. commune is very desiccation tolerant and water uptake from saturated NaCl solutions (either by direct uptake of brine or adsorption of humidity) did not enhance recultivability in long-time desiccated samples. Still, a minor part can grow under highly saline conditions. However, the salinity level, although unfavourable for the host organism, might be for parts of the heterotrophic microbiota no serious hindrance for growing in salty Mars-like environments.

  6. Differential effect of ultraviolet-B radiation on certain metabolic processes in a chromatically adapting Nostoc

    Energy Technology Data Exchange (ETDEWEB)

    Tyagi, R.; Srinivas, G.; Vyas, D.; Kumar, A.; Kumar, H.D. (Banaras Hindu Univ., Varanasi (India))

    1992-03-01

    The impact of UV-B radiation on growth, pigmentation and certain physiological processes was studied in a N{sub 2}-fixing chromatically adapting cyanobacterium, Nostoc spongiaeforme. A brownish form (phycoerythrin rich) was found to be more tolerant to UV-B than the blue-green (phycocyanin rich) form of N. spongiaeforme. Continuous exposure to UV-B (5.5 W m{sup -2}) for 90 min caused complete killing of the blue-green strain whereas the brown strain showed complete loss of survival after 180 min. Pigment content was more strongly inhibited in the blue-green strain than in the brown. Nitrogenase activity was completely abolished in both strains within 35 min of UV-B treatment. Restoration of nitrogenase occurred upon transfer to fluorescent or incandescent light after a lag of 5-6 h, suggesting fresh synthesis of nitrogenase. In vivo nitrate reductase activity was stimulated by UV-B treatment, the degree of enhancement being significantly higher in the blue-green strain. {sup 14}CO{sub 2} uptake was also completely abolished by UV-B treatment in both strains. (author).

  7. 2-Methylhopanoids are maximally produced in akinetes of Nostoc punctiforme: geobiological implications

    Science.gov (United States)

    Doughty, David M.; Hunter, Ryan C.; Summons, Roger E.; Newman, Dianne K.

    2010-01-01

    2-Methylhopanes, molecular fossils of 2-methylbacteriohopanepolyol (2-MeBHP) lipids, have been proposed as biomarkers for cyanobacteria, and by extension, oxygenic photosynthesis. However, the robustness of this interpretation is unclear, as 2-methylhopanoids occur in organisms besides cyanobacteria and their physiological functions are unknown. As a first step towards understanding the role of 2-MeBHP in cyanobacteria, we examined the expression and intercellular localization of hopanoids in the three cell types of Nostoc punctiforme: vegetative cells, akinetes, and heterocysts. Cultures in which N. punctiforme had differentiated into akinetes contained approximately 10-fold higher concentrations of 2-methylhopanoids than did cultures that contained only vegetative cells. In contrast, 2-methylhopanoids were only present at very low concentrations in heterocysts. Hopanoid production initially increased 3-fold in cells starved of nitrogen but returned to levels consistent with vegetative cells within two weeks. Vegetative and akinete cell types were separated into cytoplasmic, thylakoid, and outer membrane fractions; the increase in hopanoid expression observed in akinetes was due to a 34-fold enrichment of hopanoid content in their outer membrane relative to vegetative cells. Akinetes formed in response either to low light or phosphorus limitation, exhibited the same 2-methylhopanoid localization and concentration, demonstrating that 2-methylhopanoids are associated with the akinete cell type per se. Because akinetes are resting cells that are not photosynthetically active, 2-methylhopanoids cannot be functionally linked to oxygenic photosyntheis in N. punctiforme. PMID:19811542

  8. Metabolomic approach to optimizing and evaluating antibiotic treatment in the axenic culture of cyanobacterium Nostoc flagelliforme.

    Science.gov (United States)

    Han, Pei-pei; Jia, Shi-ru; Sun, Ying; Tan, Zhi-lei; Zhong, Cheng; Dai, Yu-jie; Tan, Ning; Shen, Shi-gang

    2014-09-01

    The application of antibiotic treatment with assistance of metabolomic approach in axenic isolation of cyanobacterium Nostoc flagelliforme was investigated. Seven antibiotics were tested at 1-100 mg L(-1), and order of tolerance of N. flagelliforme cells was obtained as kanamycin > ampicillin, tetracycline > chloromycetin, gentamicin > spectinomycin > streptomycin. Four antibiotics were selected based on differences in antibiotic sensitivity of N. flagelliforme and associated bacteria, and their effects on N. flagelliforme cells including the changes of metabolic activity with antibiotics and the metabolic recovery after removal were assessed by a metabolomic approach based on gas chromatography-mass spectrometry combined with multivariate analysis. The results showed that antibiotic treatment had affected cell metabolism as antibiotics treated cells were metabolically distinct from control cells, but the metabolic activity would be recovered via eliminating antibiotics and the sequence of metabolic recovery time needed was spectinomycin, gentamicin > ampicillin > kanamycin. The procedures of antibiotic treatment have been accordingly optimized as a consecutive treatment starting with spectinomycin, then gentamicin, ampicillin and lastly kanamycin, and proved to be highly effective in eliminating the bacteria as examined by agar plating method and light microscope examination. Our work presented a strategy to obtain axenic culture of N. flagelliforme and provided a method for evaluating and optimizing cyanobacteria purification process through diagnosing target species cellular state.

  9. A Nostoc punctiforme sugar transporter necessary to establish a Cyanobacterium-plant symbiosis.

    Science.gov (United States)

    Ekman, Martin; Picossi, Silvia; Campbell, Elsie L; Meeks, John C; Flores, Enrique

    2013-04-01

    In cyanobacteria-plant symbioses, the symbiotic nitrogen-fixing cyanobacterium has low photosynthetic activity and is supplemented by sugars provided by the plant partner. Which sugars and cyanobacterial sugar uptake mechanism(s) are involved in the symbiosis, however, is unknown. Mutants of the symbiotically competent, facultatively heterotrophic cyanobacterium Nostoc punctiforme were constructed bearing a neomycin resistance gene cassette replacing genes in a putative sugar transport gene cluster. Results of transport activity assays using (14)C-labeled fructose and glucose and tests of heterotrophic growth with these sugars enabled the identification of an ATP-binding cassette-type transporter for fructose (Frt), a major facilitator permease for glucose (GlcP), and a porin needed for the optimal uptake of both fructose and glucose. Analysis of green fluorescent protein fluorescence in strains of N. punctiforme bearing frt::gfp fusions showed high expression in vegetative cells and akinetes, variable expression in hormogonia, and no expression in heterocysts. The symbiotic efficiency of N. punctiforme sugar transport mutants was investigated by testing their ability to infect a nonvascular plant partner, the hornwort Anthoceros punctatus. Strains that were specifically unable to transport glucose did not infect the plant. These results imply a role for GlcP in establishing symbiosis under the conditions used in this work.

  10. Sorption and desorption studies of chromium(VI) from nonviable cyanobacterium Nostoc muscorum biomass

    International Nuclear Information System (INIS)

    Gupta, V.K.; Rastogi, A.

    2008-01-01

    This communication presents results pertaining to the sorptive and desorptive studies carried out on chromium(VI) removal onto nonviable freshwater cyanobacterium (Nostoc muscorum) biomass. Influence of varying the conditions for removal of chromium(VI), such as the pH of aqueous solution, the dosage of biosorbent, the contact time with the biosorbent, the temperature for the removal of chromium, the effect of light metal ions and the adsorption-desorption studies were investigated. Sorption interaction of chromium on to cyanobacterial species obeyed both the first and the second-order rate equation and the experimental data showed good fit with both the Langmuir and freundlich adsorption isotherm models. The maximum adsorption capacity was 22.92 mg/g at 25 o C and pH 3.0. The adsorption process was endothermic and the values of thermodynamic parameters of the process were calculated. Various properties of the cyanobacterium, as adsorbent, explored in the characterization part were chemical composition of the adsorbent, surface area calculation by BET method and surface functionality by FTIR. Sorption-desorption of chromium into inorganic solutions and distilled water were observed and this indicated the biosorbent could be regenerated using 0.1 M HNO 3 and EDTA with upto 80% recovery. The biosorbents were reused in five biosorption-desorption cycles without a significant loss in biosorption capacity. Thus, this study demonstrated that the cyanobacterial biomass N. muscorum could be used as an efficient biosorbent for the treatment of chromium(VI) bearing wastewater

  11. Bacterial Species and Biochemical Characteristic Investigations of Nostoc flagelliforme Concentrates during its Storage.

    Science.gov (United States)

    Yue, Lifang; Lv, Hexin; Zhen, Jing; Jiang, Shengping; Jia, Shiru; Shen, Shigang; Gao, Lu; Dai, Yujie

    2016-04-28

    Preservation of fresh algae plays an important role in algae seed subculture and aquaculture. The determination and examination of the changes of cell viability, composition, and bacterial species during storage would help to take suitable preservation methods to prolong the preservation time of fresh algae. Nostoc flagelliforme is a kind of edible cyanobacterium with important herbal and dietary values. This article investigated the changes of bacterial species and biochemical characteristics of fresh N. flagelliforme concentrate during natural storage. It was found that the viability of cells decreased along with the storage time. Fourteen bacteria strains in the algae concentrate were identified by PCR-DGGE and were grouped into four phyla, including Cyanobacteria, Firmicutes, Proteobacteria, and Bacteroidetes. Among them, Enterococcus viikkiensis may be a concern in the preservation. Eleven volatile organic compounds were identified from N. flagelliforme cells, in which geosmin could be treated as an indicator of the freshness of N. flagelliforme. The occurrence of indole compound may be an indicator of the degradation of cells.

  12. Differential effect of ultraviolet-B radiation on certain metabolic processes in a chromatically adapting Nostoc

    International Nuclear Information System (INIS)

    Tyagi, R.; Srinivas, G.; Vyas, D.; Kumar, A.; Kumar, H.D.

    1992-01-01

    The impact of UV-B radiation on growth, pigmentation and certain physiological processes was studied in a N 2 -fixing chromatically adapting cyanobacterium, Nostoc spongiaeforme. A brownish form (phycoerythrin rich) was found to be more tolerant to UV-B than the blue-green (phycocyanin rich) form of N. spongiaeforme. Continuous exposure to UV-B (5.5 W m -2 ) for 90 min caused complete killing of the blue-green strain whereas the brown strain showed complete loss of survival after 180 min. Pigment content was more strongly inhibited in the blue-green strain than in the brown. Nitrogenase activity was completely abolished in both strains within 35 min of UV-B treatment. Restoration of nitrogenase occurred upon transfer to fluorescent or incandescent light after a lag of 5-6 h, suggesting fresh synthesis of nitrogenase. In vivo nitrate reductase activity was stimulated by UV-B treatment, the degree of enhancement being significantly higher in the blue-green strain. 14 CO 2 uptake was also completely abolished by UV-B treatment in both strains. (author)

  13. Effect of light with different wavelengths on Nostoc flagelliforme cells in liquid culture.

    Science.gov (United States)

    Dai, Yu-Jie; Li, Jing; Wei, Shu-Mei; Chen, Nan; Xiao, Yu-Peng; Tan, Zhi-Lei; Jia, Shi-Ru; Yuan, Nan-Nan; Tan, Ning; Song, Yi-Jie

    2013-04-01

    The effects of lights with different wavelengths on the growth and the yield of extracellular polysaccharides of Nostoc flagelliforme cells were investigated in a liquid cultivation. N. flagelliforme cells were cultured for 16 days in 500 ml conical flasks containing BG11 culture medium under 27 micromol·m-2·s-1 of light intensity and 25 degrees C on a rotary shaker (140 rpm). The chlorophyll a, phycocyanin, allophycocyanin, and phycoerythrin contents in N. flagelliforme cells under the lights of different wavelengths were also measured. It was found that the cell biomass and the yield of polysaccharide changed with different wavelengths of light. The biomass and the yield of extracellular polysaccharides under the red or violet light were higher than those under other light colors. Chlorophyll a, phycocyanin, and allophycocyanin are the main pigments in N. flagelliforme cells. The results showed that N. flagelliforme, like other cyanobacteria, has the ability of adjusting the contents and relative ratio of its pigments with the light quality. As a conclusion, N. flagelliforme cells favor red and violet lights and perform the complementary chromatic adaptation ability to acclimate to the changes of the light quality in the environment.

  14. Preparation of desiccation-resistant aquatic-living Nostoc flagelliforme (Cyanophyceae) for potential ecological application

    Science.gov (United States)

    Gao, Xiang; Yang, Yi-Wen; Cui, Li-Juan; Zhou, De-Bao; Qiu, Bao-Sheng

    2015-01-01

    Nostoc flagelliforme is a terrestrial edible cyanobacterium that grows in arid and semi-arid steppes. The continued over-exploitation in the last century has led to a sharp decline of this resource and a severe deterioration of the steppe ecology. Liquid-cultured N. flagelliforme serves as promising algal ‘seeds’ for resource restoration. In this study, macroscopic (or visible) aquatic-living colonies (MaACs) of N. flagelliforme were developed under weak light and high nitrogen conditions. In a 24 day shake-flask culture, MaACs were propagated by about 4.5-fold in biomass without loss of their macro-morphology; at the same time, the addition of weak UV-B treatment resulted in slightly bigger MaACs. Polyvinylpyrrolidone (PVP) k30, a water-soluble polymer, was used to generate the coating around MaACs, and after full desiccation, the coated MaACs could recover their photosynthetic physiological activity when rehydrated, with 4% PVP k30 for coating being most effective. In contrast, PVP k30-coated microscopic aquatic-living colonies of N. flagelliforme and non-coated MaACs showed no resistance to full desiccation. The macroscopic morphology or structure of MaACs should be crucial for the formation of protection by PVP k30 coating. PVP k30-coated MaACs were more approaching to actual application for resource restoration. PMID:25847617

  15. Cellular and functional specificity among ferritin-like proteins in the multicellular cyanobacterium Nostoc punctiforme.

    Science.gov (United States)

    Ekman, Martin; Sandh, Gustaf; Nenninger, Anja; Oliveira, Paulo; Stensjö, Karin

    2014-03-01

    Ferritin-like proteins constitute a remarkably heterogeneous protein family, including ferritins, bacterioferritins and Dps proteins. The genome of the filamentous heterocyst-forming cyanobacterium Nostoc punctiforme encodes five ferritin-like proteins. In the present paper, we report a multidimensional characterization of these proteins. Our phylogenetic and bioinformatics analyses suggest both structural and physiological differences among the ferritin-like proteins. The expression of these five genes responded differently to hydrogen peroxide treatment, with a significantly higher rise in transcript level for Npun_F3730 as compared with the other four genes. A specific role for Npun_F3730 in the cells tolerance against hydrogen peroxide was also supported by the inactivation of Npun_F3730, Npun_R5701 and Npun_R6212; among these, only the ΔNpun_F3730 strain showed an increased sensitivity to hydrogen peroxide compared with wild type. Analysis of promoter-GFP reporter fusions of the ferritin-like genes indicated that Npun_F3730 and Npun_R5701 were expressed in all cell types of a diazotrophic culture, while Npun_F6212 was expressed specifically in heterocysts. Our study provides the first comprehensive analysis combining functional differentiation and cellular specificity within this important group of proteins in a multicellular cyanobacterium. © 2013 John Wiley & Sons Ltd and Society for Applied Microbiology.

  16. The biotechnology of hydrogen production by Nostoc flagelliforme grown under chemostat conditions

    Energy Technology Data Exchange (ETDEWEB)

    Lichtl, R.R.; Bazin, M.J.; Hall, D.O. [Div. of Life Sciences, King`s College, London Univ. (United Kingdom)

    1997-11-01

    The potential of using N{sub 2}-fixing cyanobacteria to produce hydrogen photobiologically has stimulated research on the physiology and biotechnology of species exhibiting high H{sub 2} production rates over long periods of time. In this work Nostoc flagelliforme, a terrestrial N{sub 2}-fixing cyanobacterium, has been examined to establish its physiology and potential for H{sub 2} production under controlled conditions. Cell filaments of N. flagelliforme were purified and grown in liquid culture to optimize its H{sub 2} metabolism. In batch-grown cultures the activity of nitrogenase, the key enzyme for H{sub 2} production in N{sub 2}-fixing organisms, was found to be high only during a short phase of exponential growth. A chemostat system was thus constructed for long-term experiments using continuous cultures, with the aim of exploiting the exponential growth phase. The dilution rate (D) and environmental factors, such as N{sub 2} concentration in the gas phase and temperature, significantly influenced H{sub 2} production. Cells grown continuously under the optimized conditions of D=0.022 h{sup -1}, 34 C and 5.1 kPa N{sub 2} in the gas phase exhibited H{sub 2} production rates that were more than four times higher than the maximal rates under standard batch growth conditions. (orig.)

  17. Nostopeptolide plays a governing role during cellular differentiation of the symbiotic cyanobacterium Nostoc punctiforme.

    Science.gov (United States)

    Liaimer, Anton; Helfrich, Eric J N; Hinrichs, Katrin; Guljamow, Arthur; Ishida, Keishi; Hertweck, Christian; Dittmann, Elke

    2015-02-10

    Nostoc punctiforme is a versatile cyanobacterium that can live either independently or in symbiosis with plants from distinct taxa. Chemical cues from plants and N. punctiforme were shown to stimulate or repress, respectively, the differentiation of infectious motile filaments known as hormogonia. We have used a polyketide synthase mutant that accumulates an elevated amount of hormogonia as a tool to understand the effect of secondary metabolites on cellular differentiation of N. punctiforme. Applying MALDI imaging to illustrate the reprogramming of the secondary metabolome, nostopeptolides were identified as the predominant difference in the pks2(-) mutant secretome. Subsequent differentiation assays and visualization of cell-type-specific expression of nostopeptolides via a transcriptional reporter strain provided evidence for a multifaceted role of nostopeptolides, either as an autogenic hormogonium-repressing factor or as a chemoattractant, depending on its extracellular concentration. Although nostopeptolide is constitutively expressed in the free-living state, secreted levels dynamically change before, during, and after the hormogonium differentiation phase. The metabolite was found to be strictly down-regulated in symbiosis with Gunnera manicata and Blasia pusilla, whereas other metabolites are up-regulated, as demonstrated via MALDI imaging, suggesting plants modulate the fine-balanced cross-talk network of secondary metabolites within N. punctiforme.

  18. Biosorption of Cd(II) and Zn(II) by nostoc commune: isotherm and kinetics studies

    Energy Technology Data Exchange (ETDEWEB)

    Morsy, Fatthy M. [Faculty of Science, Botany Department, Assiut University, Assiut (Egypt); Hassan, Sedky H.A. [Department of Biological Environment, Kangwon National University, Kangwon-do (Korea, Republic of); Koutb, Mostafa [Faculty of Science, Botany Department, Assiut University, Assiut (Egypt); Umm Al-Qura University, Faculty of Applied Science, Biology Department, Mecca (Saudi Arabia)

    2011-07-15

    In this study, Nostoc commune (cyanobacterium) was used as an inexpensive and efficient biosorbent for Cd(II) and Zn(II) removal from aqueous solutions. The effect of various physicochemical factors on Cd(II) and Zn(II) biosorption such as pH 2.0-7.0, initial metal concentration 0.0-300 mg/L and contact time 0-120 min were studied. Optimum pH for removal of Cd(II) and Zn(II) was 6.0, while the contact time was 30 min at room temperature. The nature of biosorbent and metal ion interaction was evaluated by infrared (IR) technique. IR analysis of bacterial biomass revealed the presence of amino, carboxyl, hydroxyl, and carbonyl groups, which are responsible for biosorption of Cd(II) and Zn (II). The maximum biosorption capacities for Cd(II) and Zn(II) biosorption by N. commune calculated from Langmuir biosorption isotherm were 126.32 and 115.41 mg/g, respectively. The biosorption isotherm for two biosorbents fitted well with Freundlich isotherm than Langmuir model with correlation coefficient (r{sup 2} < 0.99). The biosorption kinetic data were fitted well with the pseudo-second-order kinetic model. Thus, this study indicated that the N. commune is an efficient biosorbent for the removal of Cd(II) and Zn(II) from aqueous solutions. (Copyright copyright 2011 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  19. Feathermoss and epiphytic Nostoc cooperate differently: expanding the spectrum of plant–cyanobacteria symbiosis

    Energy Technology Data Exchange (ETDEWEB)

    Warshan, Denis; Espinoza, Josh L.; Stuart, Rhona; Richter, Alexander R.; Kim, Sea-Yong; Shapiro, Nicole; Woyke, Tanja; Kyripides, Nikos; Barry, Kerrie W.; Singan, Vasanth; Lindquist, Erika; Ansong, Charles K.; Purvine, Samuel O.; Brewer, Heather M.; Weyman, Philip D.; Dupont, Chris; Rasmussen, Ulla

    2017-12-31

    Dinitrogen (N2)-fixation by cyanobacteria in symbiosis with feather mosses represents the main pathway of biological N input into boreal forests. Despite its significance, little is known about the gene repertoire needed for the establishment and maintenance of the symbiosis. To determine gene acquisitions or regulatory rewiring allowing cyanobacteria to form this symbiosis, we compared closely related Nostoc strains that were either symbiosis-competent or non-competent, using a proteogenomics approach and a unique experimental setup allowing for controlled chemical and physical contact between partners. Thirty-two protein families were only in the genomes of competent strains, including some never before associated with symbiosis. We identified conserved orthologs that were differentially expressed in competent strains, including gene families involved in chemotaxis and motility, NO regulation, sulfate/phosphate transport, sugar metabolism, and glycosyl-modifying and oxidative stress-mediating exoenzymes. In contrast to other cyanobacteria-plant symbioses, the moss-cyanobacteria epiphytic symbiosis is distinct, with the symbiont retaining motility and chemotaxis, and not modulating N-fixation, photosynthesis, GS-GOGAT cycle, and heterocyst formation. Our work expands our knowledge of plant cyanobacterial symbioses, provides an interaction model of this ecologically significant symbiosis, and suggests new currencies, namely nitric oxide and aliphatic sulfonates, may be involved in establishing and maintaining this symbiosis.

  20. Evolutive and structural characterization of Nostoc commune iron-superoxide dismutase that is fit for modification.

    Science.gov (United States)

    Ma, Y; Lu, M; Li, J-Y; Qin, Y; Gong, X-G

    2012-10-04

    Superoxide dismutase (SOD) has extensive clinical applications for protecting organisms from toxic oxidation. In this study, the integrated iron-superoxide dismutase gene (fe-sod) coding sequence of Nostoc commune stain CHEN was cloned from genomic DNA and compared to sods from other reported algae. These analyses of immunology and phylogenetics indicated that this Fe-SOD is considerably homologous with SODs from lower prokaryotes (Fe-SOD or Mn-SOD) but not those from higher animals (Cu/Zn-SOD). In addition, the N. commune Fe-SOD shows 67 to 93% protein sequence identity to 10 other algal Fe-SODs (or Mn-SODs) and 69 to 93% gene sequence identity. Rare nonsynonymous substitutions imply that algal SODs are being subjected to strong natural selection. Interestingly, the N. commune Fe-SOD enzyme molecule has a compact active center that is highly conserved (38.1% of residues are absolutely conserved), and 2 loose ends localized outside the molecule and inclined to mutate (only 11.5% of residues are absolutely conserved). Based on associative analyses of evolution, structure, and function, this special phenomenon is attributed to function-dependent evolution through negative natural selection. Under strong natural selection, although the mutation is random on the gene level, the exterior region is inclined to mutate on the protein level owing to more nonsynonymous substitutions in the exterior region, which demonstrates the theoretical feasibility of modifying Fe-SOD on its ends to overcome its disadvantages in clinical applications.

  1. 3D-nuclear heat generation in PCC-charcoal filter in TAPP-3 and 4

    International Nuclear Information System (INIS)

    Kaushal, Manish; Pradhan, A.S.; Kumar, A.N.

    2006-01-01

    This paper deals with the calculations of 3D nuclear heat generation profile in the charcoal filter and subsequently the commencement time of Primary Containment Cleanup (PCC) system of 540MWe Pressurized Heavy Water Reactor (PHWR). Fuel failure is predicted due to overheating of the fuel under loss of Coolant Accident (LOCA) without Emergency Core Cooling System (LOCA without ECCS). Subsequently fission product gasses along with water vapours are released to Reactor Building (RB) atmosphere. Plate-out and water trapping mechanism stabilizes the concentration of significant fission products i.e. radioiodines in about 4 hours before being circulated through charcoal filters of Containment Cleanup system. After cleaning up the RB atmosphere, it is discharged to outside atmosphere through stack. The isotopes of radioiodine emit beta and gamma radiations. Gamma radiations are partly stopped within the charcoal and heat is generated. The part of gamma radiations escaping the bed produce heat in the adjacent beds also. PCC system can be operated, after 4 hours of LOCA, based on radioiodine concentration in RB atmosphere. During iodine removal, the iodine concentration in the charcoal filter goes through a peak value. Maximum heat is generated in the filter if PCC fans stops eventually when iodine concentration in the filter is maximum. Analysis done by TRAFIC code indicates that the system can be commenced after 7 hrs of LOCA so that desorption temperature of charcoal is not reached. Accuracy in estimating heat generation rates in charcoal helps in deciding commencement of the system after LOCA

  2. Flux coupling and transcriptional regulation within the metabolic network of the photosynthetic bacterium Synechocystis sp. PCC6803

    DEFF Research Database (Denmark)

    Montagud, Arnau; Zelezniak, Aleksej; Navarro, Emilio

    2011-01-01

    networks, surrounded by a stable core of pathways leading to biomass building blocks. This analysis identified potential bottlenecks for hydrogen and ethanol production. Integration of transcriptomic data with the Synechocystis flux coupling networks lead to identification of reporter flux coupling pairs...

  3. NADPH-Thioredoxin Reductase C Mediates the Response to Oxidative Stress and Thermotolerance in the Cyanobacterium Anabaena sp PCC7120

    NARCIS (Netherlands)

    Sanchez-Riego, Ana M.; Mata-Cabana, Alejandro; Galmozzi, CarlaV.; Florencio, Francisco J.

    2016-01-01

    NADPH-thioredoxin reductase C (NTRC) is a bimodular enzyme composed of an NADPH-thioredoxin reductase and a thiioredoxin domain extension in the same protein. In plants, NTRC has been described to be involved in the protection of the chloroplast against oxidative stress damage through reduction of

  4. Secondary structure estimation of recombinant psbH, encoding a photosynthetic membrane protein of cyanobacterium Synechocystis sp. PCC 6803

    Czech Academy of Sciences Publication Activity Database

    Štys, Dalibor

    2005-01-01

    Roč. 43, č. 3 (2005), s. 421-424 ISSN 0300-3604 Institutional research plan: CEZ:AV0Z60870520; MSM6007665808 Keywords : protein Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 0.810, year: 2005

  5. Photosynthetic Versatility in the Genome of Geitlerinema sp. PCC 9228 (Formerly Oscillatoria limnetica ?Solar Lake?), a Model Anoxygenic Photosynthetic Cyanobacterium

    OpenAIRE

    Grim, Sharon L.; Dick, Gregory J.

    2016-01-01

    Anoxygenic cyanobacteria that use sulfide as the electron donor for photosynthesis are a potentially influential but poorly constrained force on Earth’s biogeochemistry. Their versatile metabolism may have boosted primary production and nitrogen cycling in euxinic coastal margins in the Proterozoic. In addition, they represent a biological mechanism for limiting the accumulation of atmospheric oxygen, especially before the Great Oxidation Event and in the low-oxygen conditions of the Proteroz...

  6. NADPH-thioredoxin reductase C mediates the response to oxidative stress and thermotolerance in the cyanobacterium Anabaena sp. PCC7120.

    Directory of Open Access Journals (Sweden)

    ANA MARÍA SÁNCHEZ-RIEGO

    2016-08-01

    Full Text Available NTRC (NADPH-thioredoxin reductase C is a bimodular enzyme composed of an NADPH-thioredoxin reductase and a thioredoxin domain extension in the same protein. In plants, NTRC has been described to be involved in the protection of the chloroplast against oxidative stress damage through reduction of the 2-Cys peroxiredoxin (2-Cys Prx as well as through other functions related to redox enzyme regulation. In cyanobacteria, the Anabaena NTRC has been characterized in vitro, however nothing was known about its in vivo function. In order to study that, we have generated the first knockout mutant strain (∆ntrC, apart from the previously described in Arabidopsis. Detailed characterization of this strain reveals a differential sensitivity to oxidative stress treatments with respect to the wild-type Anabaena strain, including a higher level of ROS (reactive oxygen species in normal growth conditions. In the mutant strain, different oxidative stress treatments such as hydrogen peroxide, methyl-viologen or high light irradiance provoke an increase in the expression of genes related to ROS detoxification, including AnNTRC and peroxiredoxin genes, with a concomitant increase in the amount of AnNTRC and 2-Cys Prx. Moreover, the role of AnNTRC in the antioxidant response is confirmed by the observation of a pronounced overoxidation of the 2-Cys Prx and a time-delay recovery of the reduced form of this protein upon oxidative stress treatments. Our results suggest the participation of this enzyme in the peroxide detoxification in Anabaena. In addition, we describe the role of Anabaena NTRC in thermotolerance, by the appearance of high molecular mass AnNTRC complexes, showing that the mutant strain is more sensitive to high temperature treatments.

  7. Photosystem II Assembly Steps Take Place in the Thylakoid Membrane of the Cyanobacterium Synechocystis sp PCC6803

    Czech Academy of Sciences Publication Activity Database

    Sealo, T.T.; Zhang, L.; Knoppová, Jana; Komenda, Josef; Norling, B.

    2016-01-01

    Roč. 57, č. 1 (2016), s. 95-104 ISSN 0032-0781 R&D Projects: GA ČR GBP501/12/G055; GA MŠk LO1416 Institutional support: RVO:61388971 Keywords : Aqueous two-phase partitioning * Cyanobacteria * Photosystem II biogenesis Subject RIV: EE - Microbiology, Virology Impact factor: 4.760, year: 2016

  8. Chirality Matters: Synthesis and Consumption of the d-Enantiomer of Lactic Acid by Synechocystis sp. Strain PCC6803

    NARCIS (Netherlands)

    Angermayr, S.A.; van der Woude, A.D.; Correddu, D.; Kern, R.; Hagemann, M.; Hellingwerf, K.J.

    2015-01-01

    Both enantiomers of lactic acid, l-lactic acid and d-lactic acid, can be produced in a sustainable way by a photosynthetic microbial cell factory and thus from CO2, sunlight, and water. Several properties of polylactic acid (a polyester of polymerized lactic acid) depend on the controlled blend of

  9. Lipid and carotenoid cooperation-driven adaptation to light and temperature stress in Synechocystis sp. PCC6803

    Czech Academy of Sciences Publication Activity Database

    Zakar, T.; Herman, E.; Vajravel, S.; Kovács, L.; Knoppová, Jana; Komenda, Josef; Domonkos, I.; Kis, M.; Gombos, Z.; Laczko-Dobos, H.

    2017-01-01

    Roč. 1858, č. 5 (2017), s. 337-350 ISSN 0005-2728 R&D Projects: GA MŠk(CZ) LM2015055; GA MŠk(CZ) LO1416 Institutional support: RVO:61388971 Keywords : Lipid- car otenoid-protein interactions * Lipid remodeling * Xanthophylls Subject RIV: EE - Microbiology, Virology OBOR OECD: Microbiology Impact factor: 4.932, year: 2016

  10. Involvement of carotenoids in the synthesis and assembly of protein subunits of photosynthetic reaction centers of Synechocystis sp. PCC 6803

    Czech Academy of Sciences Publication Activity Database

    Sozer, O.; Komenda, Josef; Ughy, B.; Domonkos, I.; Laczkó-Dobos, H.; Malec, P.; Gombos, Z.; Kis, M.

    2010-01-01

    Roč. 51, č. 5 (2010), s. 823-835 ISSN 0032-0781 R&D Projects: GA AV ČR IAA400200801 Institutional research plan: CEZ:AV0Z50200510 Keywords : Carotenoidless mutant * crtB * Membrane protein synthesis Subject RIV: EE - Microbiology, Virology Impact factor: 4.257, year: 2010

  11. CyanoP is Involved in the Early Steps of Photosystem II Assembly in the Cyanobacterium Synechocystis sp PCC 6803

    Czech Academy of Sciences Publication Activity Database

    Knoppová, Jana; Yu, J.; Koník, P.; Nixon, P.; Komenda, Josef

    2016-01-01

    Roč. 57, č. 9 (2016), s. 1921-1931 ISSN 0032-0781 R&D Projects: GA ČR GBP501/12/G055; GA MŠk(CZ) LM2015055; GA MŠk(CZ) LO1416 Institutional support: RVO:61388971 Keywords : Cyanobacteria * CyanoP * Photosynthesis Subject RIV: EE - Microbiology, Virology Impact factor: 4.760, year: 2016

  12. Characterization of a model cyanobacterium Synechocystis sp. PCC 6803 autotrophic growth in a flat-panel photobioreactor

    Czech Academy of Sciences Publication Activity Database

    Zavřel, T.; Sinětova, M. A.; Búzová, Diana; Literáková, Petra; Červený, Jan

    2015-01-01

    Roč. 15, č. 1 (2015), s. 122-132 ISSN 1618-0240 R&D Projects: GA MŠk(CZ) ED1.1.00/02.0073; GA MŠk(CZ) EE2.3.20.0256 Institutional support: RVO:67179843 Keywords : Carbon dioxide * Exponential phase * Growth optimization * Light * Temperature Subject RIV: EH - Ecology, Behaviour Impact factor: 2.168, year: 2015

  13. SynechoNET: integrated protein-protein interaction database of a model cyanobacterium Synechocystis sp. PCC 6803

    OpenAIRE

    Kim, Woo-Yeon; Kang, Sungsoo; Kim, Byoung-Chul; Oh, Jeehyun; Cho, Seongwoong; Bhak, Jong; Choi, Jong-Soon

    2008-01-01

    Background Cyanobacteria are model organisms for studying photosynthesis, carbon and nitrogen assimilation, evolution of plant plastids, and adaptability to environmental stresses. Despite many studies on cyanobacteria, there is no web-based database of their regulatory and signaling protein-protein interaction networks to date. Description We report a database and website SynechoNET that provides predicted protein-protein interactions. SynechoNET shows cyanobacterial domain-domain interactio...

  14. Ecophysiology of gelatinous Nostoc colonies: unprecedented slow growth and survival in resource-poor and harsh environments.

    Science.gov (United States)

    Sand-Jensen, Kaj

    2014-07-01

    The cyanobacterial genus Nostoc includes several species forming centimetre-large gelatinous colonies in nutrient-poor freshwaters and harsh semi-terrestrial environments with extended drought or freezing. These Nostoc species have filaments with normal photosynthetic cells and N2-fixing heterocysts embedded in an extensive gelatinous matrix of polysaccharides and many other organic substances providing biological and environmental protection. Large colony size imposes constraints on the use of external resources and the gelatinous matrix represents extra costs and reduced growth rates. The objective of this review is to evaluate the mechanisms behind the low rates of growth and mortality, protection against environmental hazards and the persistence and longevity of gelatinous Nostoc colonies, and their ability to economize with highly limiting resources. Simple models predict the decline in uptake of dissolved inorganic carbon (DIC) and a decline in the growth rate of spherical freshwater colonies of N. pruniforme and N. zetterstedtii and sheet-like colonies of N. commune in response to a thicker diffusion boundary layer, lower external DIC concentration and higher organic carbon mass per surface area (CMA) of the colony. Measured growth rates of N. commune and N. pruniforme at high DIC availability comply with general empirical predictions of maximum growth rate (i.e. doubling time 10-14 d) as functions of CMA for marine macroalgae and as functions of tissue thickness for aquatic and terrestrial plants, while extremely low growth rates of N. zetterstedtii (i.e. doubling time 2-3 years) are 10-fold lower than model predictions, either because of very low ambient DIC and/or an extremely costly colony matrix. DIC uptake is limited by diffusion at low concentrations for all species, although they exhibit efficient HCO3(-) uptake, accumulation of respiratory DIC within the colonies and very low CO2 compensation points. Long light paths and light attenuation by

  15. Effect of UV-B and high visual radiation on photosynthesis in freshwater (nostoc spongiaeforme) and marine (Phormidium corium) cyanobacteria.

    Science.gov (United States)

    Bhandari, Rupali; Sharma, Prabhat Kumar

    2007-08-01

    Human activity is causing depletion of ozone in stratosphere, resulting in increased UV-B radiation and global warming. However, impact of these climatic changes on the aquatic organism (especially marine) is not fully understood. Here, we have studied the effect of excess UV-B and visible radiation on photosynthetic pigments, fatty acids content, lipid peroxidation, nitrogen content, nitrogen reductase activity and membrane proteins, induction of mycosporine-like amino acids (MAAs) and antioxidant enzymes superoxide dismutase (SOD) and ascorbate peroxidase (APX) in freshwater (Nostoc spongiaeform) and marine (Phormidium corium) cyanobacteria. UV-B treatment resulted in an increase in photosynthetic pigments in Nostoc and decrease in Phormidium, but high light treatment caused photobleaching of most of the pigments in both the species. Unsaturation level of fatty acids of both total and glycolipids remained unchanged in both the cyanobacteria, as a result of UV-B and high light treatments. Saturated fatty acids of total and glycolipids declined slightly in Nostoc by both the treatments. but remained unchanged in Phormidium. No changes in the unsaturated lipid content in our study probably suggested adaptation of the organism to the treatments. However, both treatments resulted in peroxidation of membrane lipids, indicating oxidative damage to lipids without any change in the level of unsaturation of fatty acid in the cell membrane. Qualitative and quantitative changes were observed in membrane protein profile due to the treatments. Cyanobacteria were able to synthesize MAAs in response to the UV-B treatment. Both treatments also increased the activities of SOD and APX. In conclusion, the study demonstrated induction of antioxidants such as SOD and APX under visible light treatment and screening pigment (MAAs) under UV-B treatment, which might protect the cyanobacteria from oxidative damage caused by high light and UV-B radiation.

  16. ORF Sequence: NC_000911 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available Synechocystis sp. PCC 6803] MYFLLVTLVILVFPLLSIALEWTTSGNSQALVDVLARWFVFWGVGVRLFLAGVVQITKPSFTAEKILGVQSQDSLILVKELGIGNLAIASVALGSIFVNAWVLGAALAGGIFYLLAGINHILQPERNAKENYAMATDLFLGLLLGGILFFAWQP

  17. Protein (Cyanobacteria): 359322 [PGDBj - Ortholog DB

    Lifescience Database Archive (English)

    Full Text Available ynechocystis sp. PCC 6803 MYFLLVTLVILVFPLLSIALEWTTSGNSQALVDVLARWFVFWGVGVRLFLAGVVQITKPSFTAEKILGVQSQDSLILVKELGIGNLAIASVALGSIFVNAWVLGAALAGGIFYLLAGINHILQPERNAKENYAMATDLFLGLLLGGILFFAWQP ...

  18. Protein (Cyanobacteria): 504930526 [PGDBj - Ortholog DB

    Lifescience Database Archive (English)

    Full Text Available hetical protein Rivularia sp. PCC 7116 MAEDNNLTNNSATNISSESQTLNKDIEELVTRQAKAWENADSEAIIADFAENGAFIAPGTSLKGKADIKKAAEDYFKEFTDTKVKITRIFSDGKEGGVEWTWSDKNKKTGEKSLIDDAIIFEIKDGKIIYWREYFDKQTVSS

  19. Protein (Cyanobacteria): 518320325 [PGDBj - Ortholog DB

    Lifescience Database Archive (English)

    Full Text Available ... hypothetical protein Calothrix sp. PCC 7103 MDYVHPFQMELHKLESMIVHVQYADIKEVDKTLASNDAVSTQAVGEEGGTKVSTRALGEEGGNILTTYAVGEEGGNILTTYAVGEEGGDKVTTQAVGEEGGTRVTTYAVGEEGGGRVTTKAVGEEGGSIIRR

  20. Protein (Cyanobacteria): 515516403 [PGDBj - Ortholog DB

    Lifescience Database Archive (English)

    Full Text Available hypothetical protein Anabaena sp. PCC 7108 MTVRFLLDSNIISEPSRPIPNIQVLDQLNRYRSEVAIASVVVHEILYGCWRLPPSKRKDSLWKYIQDSVLNLPVFDYNLNAAKWHAQERARLSKIGKTPAFIDGQIASIAFCNDLILVTNNVADFQDFQDLVIENWFI

  1. Protein (Cyanobacteria): 208270 [PGDBj - Ortholog DB

    Lifescience Database Archive (English)

    Full Text Available transferase Halothece sp. PCC 7418 MKIVIARDFNDFARCIMIRTQVFVMEQGISAEIETDEWENHSTHYLAGDGEKALATARSRLINNQTAKIERVAVLKEARSQGVGTELMRYILQEIHSYSNIQTIKLGSQNSAIPFYEKLGFQVIGEEYLDAGIPHHLMMQRINT ...

  2. Protein (Cyanobacteria): 69941 [PGDBj - Ortholog DB

    Lifescience Database Archive (English)

    Full Text Available l regulator Rivularia sp. PCC 7116 MNSLKNKPLDPVNHAGFLIWQVANNWEKQINNELKEFGLNQAEYFHLVSLFWLLENQEEVTQTEIARFADTIPMNTSKIMTKFEKKGLITRVAGSDSRSKSLCITESGEQIAIQATARLSRLSEQFFDKDDDNNFLNYLKYLKTK ...

  3. Draft genome sequences of six neonatal meningitis-causing escherichia coli isolates (SP-4, SP-5, SP-13, SP-16, SP-46, and SP-65)

    Science.gov (United States)

    Neonatal meningitis Escherichia coli isolates (SP-4, SP-5, SP-13, SP-16, SP-46, and SP-65) were recovered from infants in the Netherlands from 1989 to 1997. Here, we report the draft genome sequences for these six E. coli isolates, which are currently being used to validate food safety processing te...

  4. Role of Phosphate Transport System Component PstB1 in Phosphate Internalization by Nostoc punctiforme.

    Science.gov (United States)

    Hudek, L; Premachandra, D; Webster, W A J; Bräu, L

    2016-11-01

    In bacteria, limited phosphate availability promotes the synthesis of active uptake systems, such as the Pst phosphate transport system. To understand the mechanisms that facilitate phosphate accumulation in the cyanobacterium Nostoc punctiforme, phosphate transport systems were identified, revealing a redundancy of Pst phosphate uptake systems that exists across three distinct operons. Four separate PstB system components were identified. pstB1 was determined to be a suitable target for creating phenotypic mutations that could result in the accumulation of excessive levels of phosphate through its overexpression or in a reduction of the capacity to accumulate phosphate through its deletion. Using quantitative real-time PCR (qPCR), it was determined that pstB1 mRNA levels increased significantly over 64 h in cells cultured in 0 mM added phosphate and decreased significantly in cells exposed to high (12.8 mM) phosphate concentrations compared to the level in cells cultured under normal (0.8 mM) conditions. Possible compensation for the loss of PstB1 was observed when pstB2, pstB3, and pstB4 mRNA levels increased, particularly in cells starved of phosphate. The overexpression of pstB1 increased phosphate uptake by N. punctiforme and was shown to functionally complement the loss of PstB in E. coli PstB knockout (PstB - ) mutants. The knockout of pstB1 in N. punctiforme did not have a significant effect on cellular phosphate accumulation or growth for the most part, which is attributed to the compensation for the loss of PstB1 by alterations in the pstB2, pstB3, and pstB4 mRNA levels. This study provides novel in vivo evidence that PstB1 plays a functional role in phosphate uptake in N. punctiforme IMPORTANCE: Cyanobacteria have been evolving over 3.5 billion years and have become highly adept at growing under limiting nutrient levels. Phosphate is crucial for the survival and prosperity of all organisms. In bacteria, limited phosphate availability promotes the

  5. Preparation of desiccation-resistant aquatic-living Nostoc flagelliforme (Cyanophyceae) for potential ecological application.

    Science.gov (United States)

    Gao, Xiang; Yang, Yi-Wen; Cui, Li-Juan; Zhou, De-Bao; Qiu, Bao-Sheng

    2015-11-01

    Nostoc flagelliforme is a terrestrial edible cyanobacterium that grows in arid and semi-arid steppes. The continued over-exploitation in the last century has led to a sharp decline of this resource and a severe deterioration of the steppe ecology. Liquid-cultured N. flagelliforme serves as promising algal 'seeds' for resource restoration. In this study, macroscopic (or visible) aquatic-living colonies (MaACs) of N. flagelliforme were developed under weak light and high nitrogen conditions. In a 24 day shake-flask culture, MaACs were propagated by about 4.5-fold in biomass without loss of their macro-morphology; at the same time, the addition of weak UV-B treatment resulted in slightly bigger MaACs. Polyvinylpyrrolidone (PVP) k30, a water-soluble polymer, was used to generate the coating around MaACs, and after full desiccation, the coated MaACs could recover their photosynthetic physiological activity when rehydrated, with 4% PVP k30 for coating being most effective. In contrast, PVP k30-coated microscopic aquatic-living colonies of N. flagelliforme and non-coated MaACs showed no resistance to full desiccation. The macroscopic morphology or structure of MaACs should be crucial for the formation of protection by PVP k30 coating. PVP k30-coated MaACs were more approaching to actual application for resource restoration. © 2015 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  6. Characterization of the hupSL promoter activity in Nostoc punctiforme ATCC 29133

    Science.gov (United States)

    2009-01-01

    Background In cyanobacteria three enzymes are directly involved in the hydrogen metabolism; a nitrogenase that produces molecular hydrogen, H2, as a by-product of nitrogen fixation, an uptake hydrogenase that recaptures H2 and oxidize it, and a bidirectional hydrogenase that can both oxidize and produce H2.Nostoc punctiforme ATCC 29133 is a filamentous dinitrogen fixing cyanobacterium containing a nitrogenase and an uptake hydrogenase but no bidirectional hydrogenase. Generally, little is known about the transcriptional regulation of the cyanobacterial uptake hydrogenases. In this study gel shift assays showed that NtcA has a specific affinity to a region of the hupSL promoter containing a predicted NtcA binding site. The predicted NtcA binding site is centred at 258.5 bp upstream the transcription start point (tsp). To further investigate the hupSL promoter, truncated versions of the hupSL promoter were fused to either gfp or luxAB, encoding the reporter proteins Green Fluorescent Protein and Luciferase, respectively. Results Interestingly, all hupsSL promoter deletion constructs showed heterocyst specific expression. Unexpectedly the shortest promoter fragment, a fragment covering 57 bp upstream and 258 bp downstream the tsp, exhibited the highest promoter activity. Deletion of the NtcA binding site neither affected the expression to any larger extent nor the heterocyst specificity. Conclusion Obtained data suggest that the hupSL promoter in N. punctiforme is not strictly dependent on the upstream NtcA cis element and that the shortest promoter fragment (-57 to tsp) is enough for a high and heterocyst specific expression of hupSL. This is highly interesting because it indicates that the information that determines heterocyst specific gene expression might be confined to this short sequence or in the downstream untranslated leader sequence. PMID:19284581

  7. Hopanoids play a role in stress tolerance and nutrient storage in the cyanobacterium Nostoc punctiforme.

    Science.gov (United States)

    Ricci, J N; Morton, R; Kulkarni, G; Summers, M L; Newman, D K

    2017-01-01

    Hopanes are abundant in ancient sedimentary rocks at discrete intervals in Earth history, yet interpreting their significance in the geologic record is complicated by our incomplete knowledge of what their progenitors, hopanoids, do in modern cells. To date, few studies have addressed the breadth of diversity of physiological functions of these lipids and whether those functions are conserved across the hopanoid-producing bacterial phyla. Here, we generated mutants in the filamentous cyanobacterium, Nostoc punctiforme, that are unable to make all hopanoids (shc) or 2-methylhopanoids (hpnP). While the absence of hopanoids impedes growth of vegetative cells at high temperature, the shc mutant grows faster at low temperature. This finding is consistent with hopanoids acting as membrane rigidifiers, a function shared by other hopanoid-producing phyla. Apart from impacting fitness under temperature stress, hopanoids are dispensable for vegetative cells under other stress conditions. However, hopanoids are required for stress tolerance in akinetes, a resting survival cell type. While 2-methylated hopanoids do not appear to contribute to any stress phenotype, total hopanoids and to a lesser extent 2-methylhopanoids were found to promote the formation of cyanophycin granules in akinetes. Finally, although hopanoids support symbiotic interactions between Alphaproteobacteria and plants, they do not appear to facilitate symbiosis between N. punctiforme and the hornwort Anthoceros punctatus. Collectively, these findings support interpreting hopanes as general environmental stress biomarkers. If hopanoid-mediated enhancement of nitrogen-rich storage products turns out to be a conserved phenomenon in other organisms, a better understanding of this relationship may help us parse the enrichment of 2-methylhopanes in the rock record during episodes of disrupted nutrient cycling. © 2016 John Wiley & Sons Ltd.

  8. Molecular and cellular characterisation of the zinc uptake (Znu) system of Nostoc punctiforme.

    Science.gov (United States)

    Hudek, Lee; Pearson, Leanne A; Michalczyk, Agnes; Neilan, Brett A; Ackland, M Leigh

    2013-11-01

    Metal homoeostasis in cyanobacteria is based on uptake and export systems that are controlled by their own regulators. This study characterises the zinc uptake (Znu) system in Nostoc punctiforme. The system was found to comprise of three subunits in an ACB operon: a Zn(2+)-binding protein (ZnuA18), a transmembrane domain (ZnuB) and an ATPase (ZnuC). These proteins are encoded within the znu operon regulated by a zinc uptake transcription repressor (Zur). Interestingly, a second Zn(2+)-binding protein (ZnuA08) was also identified at a distal genomic location. Interactions between components of the ZnuACB system were investigated using knockouts of the individual genes. The znuA08(-), znuA18(-), znuB(-) and znuC(-) mutants displayed overall reduced znuACB transcript levels, suggesting that all system components are required for normal expression of znu genes. Zinc uptake assays in the Zn(2+)-binding protein mutant strains showed that the disruption of znuA18 had a greater negative effect on zinc uptake than disruption of znuA08. Complementation studies in Escherichia coli indicated that both znuA08 and znuA18 were able to restore zinc uptake in a znuA(-) mutant, with znuA18 permitting the highest zinc uptake rate. The N. punctiforme zur was also able to complement the E. coli zur(-) mutant. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  9. Biochemical characterization of an L-tryptophan dehydrogenase from the photoautotrophic cyanobacterium Nostoc punctiforme.

    Science.gov (United States)

    Ogura, Ryutaro; Wakamatsu, Taisuke; Mutaguchi, Yuta; Doi, Katsumi; Ohshima, Toshihisa

    2014-06-10

    An NAD(+)-dependent l-tryptophan dehydrogenase from Nostoc punctiforme NIES-2108 (NpTrpDH) was cloned and overexpressed in Escherichia coli. The recombinant NpTrpDH with a C-terminal His6-tag was purified to homogeneity using a Ni-NTA agarose column, and was found to be a homodimer with a molecular mass of 76.1kDa. The enzyme required NAD(+) and NADH as cofactors for oxidative deamination and reductive amination, respectively, but not NADP(+) or NADPH. l-Trp was the preferred substrate for deamination, though l-Phe was deaminated at a much lower rate. The enzyme exclusively aminated 3-indolepyruvate; phenylpyruvate was inert. The pH optima for the deamination of l-Trp and amination of 3-indolpyruvate were 11.0 and 7.5, respectively. For deamination of l-Trp, maximum enzymatic activity was observed at 45°C. NpTrpDH retained more than 80% of its activity after incubation for 30min at pHs ranging from 5.0 to 11.5 or incubation for 10min at temperatures up to 40°C. Unlike l-Trp dehydrogenases from higher plants, NpTrpDH activity was not activated by metal ions. Typical Michaelis-Menten kinetics were observed for NAD(+) and l-Trp for oxidative deamination, but with reductive amination there was marked substrate inhibition by 3-indolepyruvate. NMR analysis of the hydrogen transfer from the C4 position of the nicotinamide moiety of NADH showed that NpTrpDH has a pro-S (B-type) stereospecificity similar to the Glu/Leu/Phe/Val dehydrogenase family. Copyright © 2014 Elsevier Inc. All rights reserved.

  10. Sorption and desorption studies of chromium(VI) from nonviable cyanobacterium Nostoc muscorum biomass

    Energy Technology Data Exchange (ETDEWEB)

    Gupta, V.K. [Department of Chemistry, Indian Institute of Technology Roorkee, Roorkee 247667 (India)], E-mail: vinodfcy@iitr.ernet.in; Rastogi, A. [Department of Chemistry, Indian Institute of Technology Roorkee, Roorkee 247667 (India)

    2008-06-15

    This communication presents results pertaining to the sorptive and desorptive studies carried out on chromium(VI) removal onto nonviable freshwater cyanobacterium (Nostoc muscorum) biomass. Influence of varying the conditions for removal of chromium(VI), such as the pH of aqueous solution, the dosage of biosorbent, the contact time with the biosorbent, the temperature for the removal of chromium, the effect of light metal ions and the adsorption-desorption studies were investigated. Sorption interaction of chromium on to cyanobacterial species obeyed both the first and the second-order rate equation and the experimental data showed good fit with both the Langmuir and freundlich adsorption isotherm models. The maximum adsorption capacity was 22.92 mg/g at 25 {sup o}C and pH 3.0. The adsorption process was endothermic and the values of thermodynamic parameters of the process were calculated. Various properties of the cyanobacterium, as adsorbent, explored in the characterization part were chemical composition of the adsorbent, surface area calculation by BET method and surface functionality by FTIR. Sorption-desorption of chromium into inorganic solutions and distilled water were observed and this indicated the biosorbent could be regenerated using 0.1 M HNO{sub 3} and EDTA with upto 80% recovery. The biosorbents were reused in five biosorption-desorption cycles without a significant loss in biosorption capacity. Thus, this study demonstrated that the cyanobacterial biomass N. muscorum could be used as an efficient biosorbent for the treatment of chromium(VI) bearing wastewater.

  11. Mutational studies of putative biosynthetic genes for the cyanobacterial sunscreen scytonemin in Nostoc punctiforme ATCC 29133

    Directory of Open Access Journals (Sweden)

    Daniela eFerreira

    2016-05-01

    Full Text Available The heterocyclic indole-alkaloid scytonemin is a sunscreen found exclusively among cyanobacteria. An 18-gene cluster is responsible for scytonemin production in Nostoc punctiforme ATCC 29133. The upstream genes scyABCDEF in the cluster are proposed to be responsible for scytonemin biosynthesis from aromatic amino acid substrates. In vitro studies of ScyA, ScyB and ScyC proved that these enzymes indeed catalyze initial pathway reactions. Here we characterize the role of ScyD, ScyE and ScyF, which were logically predicted to be responsible for late biosynthetic steps, in the biological context of N. punctiforme. In-frame deletion mutants of each were constructed (∆scyD, ∆scyE and ∆scyF and their phenotypes studied. Expectedly, ∆scyE presents a scytoneminless phenotype, but no accumulation of the predicted intermediaries. Surprisingly, ∆scyD retains scytonemin production, implying that it is not required for biosynthesis. Indeed, scyD presents an interesting evolutionary paradox: it likely originated in a duplication event from scyE, and unlike other genes in the operon, it has not been subjected to purifying selection. This would suggest that it is a pseudogene, and yet scyD is highly conserved in the scytonemin operon of cyanobacteria. ∆scyF also retains scytonemin production, albeit exhibiting a reduction of the production yield compared with the wild-type. This indicates that ScyF is not essential but may play an adjuvant role for scytonemin synthesis. Altogether, our findings suggest that these downstream genes are not responsible, as expected, for the late steps of scytonemin synthesis and we must look for those functions elsewhere. These findings are particularly important for biotechnological production of this sunscreen through heterologous expression of its genes in more tractable organisms.

  12. Characterization of the hupSL promoter activity in Nostoc punctiforme ATCC 29133

    Directory of Open Access Journals (Sweden)

    Lindberg Pia

    2009-03-01

    Full Text Available Abstract Background In cyanobacteria three enzymes are directly involved in the hydrogen metabolism; a nitrogenase that produces molecular hydrogen, H2, as a by-product of nitrogen fixation, an uptake hydrogenase that recaptures H2 and oxidize it, and a bidirectional hydrogenase that can both oxidize and produce H2.Nostoc punctiforme ATCC 29133 is a filamentous dinitrogen fixing cyanobacterium containing a nitrogenase and an uptake hydrogenase but no bidirectional hydrogenase. Generally, little is known about the transcriptional regulation of the cyanobacterial uptake hydrogenases. In this study gel shift assays showed that NtcA has a specific affinity to a region of the hupSL promoter containing a predicted NtcA binding site. The predicted NtcA binding site is centred at 258.5 bp upstream the transcription start point (tsp. To further investigate the hupSL promoter, truncated versions of the hupSL promoter were fused to either gfp or luxAB, encoding the reporter proteins Green Fluorescent Protein and Luciferase, respectively. Results Interestingly, all hupsSL promoter deletion constructs showed heterocyst specific expression. Unexpectedly the shortest promoter fragment, a fragment covering 57 bp upstream and 258 bp downstream the tsp, exhibited the highest promoter activity. Deletion of the NtcA binding site neither affected the expression to any larger extent nor the heterocyst specificity. Conclusion Obtained data suggest that the hupSL promoter in N. punctiforme is not strictly dependent on the upstream NtcA cis element and that the shortest promoter fragment (-57 to tsp is enough for a high and heterocyst specific expression of hupSL. This is highly interesting because it indicates that the information that determines heterocyst specific gene expression might be confined to this short sequence or in the downstream untranslated leader sequence.

  13. Peroxide reduction by a metal-dependent catalase in Nostoc punctiforme (cyanobacteria).

    Science.gov (United States)

    Hudek, L; Torriero, A A J; Michalczyk, A A; Neilan, B A; Ackland, M L; Bräu, Lambert

    2017-05-01

    This study investigated the role of a novel metal-dependent catalase (Npun_R4582) that reduces hydrogen peroxide in the cyanobacterium Nostoc punctiforme. Quantitative real-time PCR showed that npun_R4582 relative mRNA levels were upregulated by over 16-fold in cells treated with either 2 μM added Co, 0.5 μM added Cu, 500 μM Mn, 1 μM Ni, or 18 μM Zn. For cells treated with 60 μM H 2 O 2 , no significant alteration in Npun_R4582 relative mRNA levels was detected, while in cells treated with Co, Cu, Mn, Ni, or Zn and 60 μM peroxide, relative mRNA levels were generally above control or peroxide only treated cells. Disruption or overexpression of npun_R4582 altered sensitivity to cells exposed to 60 μM H 2 O 2 and metals for treatments beyond the highest viable concentrations, or in a mixed metal solution for Npun_R4582 - cells. Moreover, overexpression of npun_R4582 increased cellular peroxidase activity in comparison with wild-type and Npun_R4582 - cells, and reduced peroxide levels by over 50%. The addition of cobalt, manganese, nickel, and zinc increased the capacity of Npun_R4582 to reduce the rate or total levels of peroxide produced by cells growing under photooxidative conditions. The work presented confirms the function of NpunR4582 as a catalase and provides insights as to how cells reduce potentially lethal peroxide levels produced by photosynthesis. The findings also show how trace elements play crucial roles as enzymatic cofactors and how the role of Npun_R4582 in hydrogen peroxide breakdown is dependent on the type of metal and the level available to cells.

  14. Cytotoxicity and Secondary Metabolites Production in Terrestrial Nostoc Strains, Originating From Different Climatic/Geographic Regions and Habitats: Is Their Cytotoxicity Environmentally Dependent?

    Czech Academy of Sciences Publication Activity Database

    Hrouzek, Pavel; Tomek, P.; Lukešová, Alena; Urban, J.; Voloshko, L.; Pushparaj, B.; Ventura, S.; Lukavský, Jaromír; Štys, D.; Kopecký, J.

    2011-01-01

    Roč. 26, č. 4 (2011), s. 345-358 ISSN 1520-4081 Institutional research plan: CEZ:AV0Z60050516; CEZ:AV0Z60660521; CEZ:AV0Z50200510 Keywords : cytotoxicity * cyanobacteria * nostoc Subject RIV: EE - Microbiology, Virology Impact factor: 2.407, year: 2011

  15. Metabolic adaptation, a specialized leaf organ structure and vascular responses to diurnal N2 fixation by nostoc azollae sustain the astonishing productivity of azolla ferns without nitrogen fertilizer

    NARCIS (Netherlands)

    Brouwer, Paul; Bräutigam, Andrea; Buijs, Valerie A.; Tazelaar, Anne O.E.; van der Werf, Adrie; Schlüter, Urte; Reichart, Gert-Jan; Bolger, Anthony; Usadel, Björn; Weber, Andreas P.M.; Schluepmann, Henriette

    2017-01-01

    Sustainable agriculture demands reduced input of man-made nitrogen (N) fertilizer, yet N2 fixation limits the productivity of crops with heterotrophic diazotrophic bacterial symbionts. We investigated floating ferns from the genus Azolla that host phototrophic diazotrophic Nostoc azollae in leaf

  16. Influence of a non-copper algicide on the cyanobacterium, Nostoc spongiaeforme, and the green alga, Hydrodictyon reticulatum, in field and laboratory experiments

    Science.gov (United States)

    Cyanobacteria grow in California rice fields where they form large mats that may smoother seedlings or cause them to dislodge, resulting in yield loss. The most troublesome species is Nostoc spongiaeforme. It is very difficult to control using currently accepted methods, i.e., aerial applications of...

  17. Phylogeny of Indonesian Nostoc (Cyanobac teria Isolated from Paddy Fields as Inferred from Partial Se quence of 16S rRNA Gene

    Directory of Open Access Journals (Sweden)

    Dian Hendrayanti

    2012-12-01

    Full Text Available In order to collect Indonesian Nostoc, isolation of soil microflora from several paddy fields in West Java, Bali, andSouth Celebes was carried out. Fast-growing isolates of Nostoc were selected to describe and perform molecular identification using partial sequences of 16S rRNA. The results showed that partial sequences of 16S rRNA could not resolve the phylogeny of the isolates. However, it supported the morphological studies that recognize isolates as different species of Nostoc. Potential use of Nostoc as a nitrogen source for paddy growth was carried out using six strains as single inoculums. A total biomass of 2 g (fresh weight for each strain was inoculated, respectively, into the pot planted with three paddy plants. This experiment was conducted in the green house for 115 days. Statistical analyses (ANOVA; α = 0.05 showed that of six strains tested in this study, only strain GIA13a had influence on the augmentation of root length and the total number of filled grains.

  18. Flexible dynamic operation of solar-integrated power plant with solvent based post-combustion carbon capture (PCC) process

    International Nuclear Information System (INIS)

    Qadir, Abdul; Sharma, Manish; Parvareh, Forough; Khalilpour, Rajab; Abbas, Ali

    2015-01-01

    Highlights: • Flexible operation of power and PCC plant may significantly increase operational revenue. • Higher optimal carbon capture rates observed with solar thermal energy input. • Solar thermal repowering of the power plant provides highest net revenue. • Constant optimal capture rate observed for one of the flexible operation cases. • Up to 42% higher revenue generation observed between two cases with solar input. - Abstract: This paper examines flexible operation of solvent-based post-combustion carbon capture (PCC) for the reduction of power plant carbon emissions while minimizing revenue loss due to the reduced power plant electricity output. The study is conducted using a model superstructure enveloping three plants; a power plant, a PCC plant and a solar thermal field where the power plant and PCC plant are operated flexibly under the influence of hourly electricity market and weather conditions. Reduced (surrogate) models for the reboiler duty and auxiliary power requirement for the carbon capture plant are generated and applied to simulate and compare four cases, (A) power plant with PCC, (B) power plant with solar assisted PCC, (C) power plant with PCC and solar repowering – variable net electricity output and (D) power plant with PCC and solar repowering – fixed net electricity output. Such analyses are conducted under dynamic conditions including power plant part-load operation while varying the capture rate to optimize the revenue of the power plant. Each case was simulated with a lower carbon price of $25/tonne-CO 2 and a higher price of $50/tonne-CO 2 . The comparison of cases B–D found that optimal revenue generation for case C can be up to 42% higher than that of solar-assisted PCC (case B). Case C is found to be the most profitable with the lowest carbon emissions intensity and is found to exhibit a constant capture rate for both carbon prices. The optimal revenue for case D is slightly lower than case C for the lower carbon

  19. Establishment of an isolation method of Nostoc commune cells free from extracellular polysaccharides (EPS using Percoll centrifugation

    Directory of Open Access Journals (Sweden)

    Makiko Kosugi

    2012-11-01

    Full Text Available The terrestrial cyanobacterium Nostoc commune Vaucher ex Bornet et Flahault occurs worldwide, including in Japan and Antarctica. N. commune has a large amount of extracellular polysaccharides (EPS that hold moisture and protect the cells and at the same time accumulate light-blocking substances which is believed to play an important part in adaptation to a severe environment. To evaluate the photoadaptation processes in N. commune and clarify the role(s of EPS under ambient environmental condition at Antarctica, separation of cells from EPS is necessary. High yield is a prerequisite for the use of only small amount of natural N. commune from Antarctica. For this purpose, we developed a separation method by improving the Percoll density gradient centrifugation method using an EPS-coated field-grown Nostoc population. We established the most suitable condition to separate naked cells from EPS at high yield retaining high photosynthetic activity. The method is composed of centrifugation of cell homogenated N. commune in 10% (v/v Percoll to separate cells efficiently from EPS followed by fractionating centrifugation to remove impurities using the gradient of Percoll (80% and 50%, v/v.

  20. Comparative transcriptomics with a motility-deficient mutant leads to identification of a novel polysaccharide secretion system in Nostoc punctiforme.

    Science.gov (United States)

    Risser, Douglas D; Meeks, John C

    2013-02-01

    Many filamentous cyanobacteria are capable of gliding motility by an undefined mechanism. Within the heterocyst-forming clades, some strains, such as the Nostoc spp. and Fisherella spp., are motile only as specialized filaments termed hormogonia. Here we report on the phenotype of inactivation of a methyl-accepting chemotaxis-like protein in Nostoc punctiforme, designated HmpD. The gene hmpD was found to be essential for hormogonium development, motility and polysaccharide secretion. Comparative global transcriptional profiling of the ΔhmpD strain demonstrated that HmpD has a profound effect on the transcriptional programme of hormogonium development, influencing approximately half of the genes differentially transcribed during differentiation. Utilizing this transcriptomic data, we identified a gene locus, designated here as hps, that appears to encode for a novel polysaccharide secretion system. Transcripts for the genes in the hps locus are upregulated in two steps, with the second step dependent on HmpD. Deletion of hpsA, hpsBCD or hpsEFG resulted in the complete loss of motility and polysaccharide secretion, similar to deletion of hmpD. Genes in the hps locus are highly conserved in the filamentous cyanobacteria, but generally absent in unicellular strains, implying a common mechanism of motility unique to the filamentous cyanobacteria. © 2012 Blackwell Publishing Ltd.

  1. Short Communication: Effects of temperature on growth, pigment composition and protein content of an Antarctic Cyanobacterium Nostoc commune

    Directory of Open Access Journals (Sweden)

    RANJANA TRIPATHI

    2012-11-01

    Full Text Available Tripathi R, Dhuldhaj UP, Singh S. 2012. Short Communication: Effects of temperature on growth, pigment composition and protein content of an Antarctic Cyanobacterium Nostoc commune. Nusantara Bioscience 4: 134-137. Effect of temperature variation on biomass accumulation, pigment composition and protein content were studied for the cyanobacterium Nostoc commune, isolated from Antarctica. Results confirmed the psychrotrophic behavior (optimum growth temperature 25◦C of the cyanobacterium. Low temperature increased the duration of lag phase and exponential growth phase. Maximum increase in biomass was recorded on 24th day at 25◦C and on 12th day at 50C. The downshift from 25 to 5◦C had almost negligible effect on chl a content. Maximal protein content was recorded for cultures growing at 50C on 12th day. The carotenoids/chl a ratio was maximum (2.48 at 50C on 9th day. It remained almost constant for cultures growing at 5 and 350C. There was an induction in protein synthesis following downshift in temperature from 25 to 5◦C.

  2. Fungal lectin of Peltigera canina induces chemotropism of compatible Nostoc cells by constriction-relaxation pulses of cyanobiont cytoskeleton.

    Science.gov (United States)

    Díaz, Eva Maria; Vicente-Manzanares, Miguel; Sacristan, Mara; Vicente, Carlos; Legaz, Maria-Estrella

    2011-10-01

    A glycosylated arginase acting as a fungal lectin from Peltigera canina is able to produce recruitment of cyanobiont Nostoc cells and their adhesion to the hyphal surface. This implies that the cyanobiont would develop organelles to motility towards the chemoattractant. However when visualized by transmission electron microscopy, Nostoc cells recently isolated from P. canina thallus do not reveal any motile, superficial organelles, although their surface was covered by small spindles and serrated layer related to gliding. The use of S-(3,4-dichlorobenzyl)isothiourea, blebbistatin, phalloidin and latrunculin A provide circumstantial evidence that actin microfilaments rather than MreB, the actin-like protein from prokaryota, and, probably, an ATPase which develops contractile function similar to that of myosin II, are involved in cell motility. These experimental facts, the absence of superficial elements (fimbriae, pili or flagellum) related to cell movement, and the appearance of sunken cells during of after movement verified by scanning electron microscopy, support the hypothesis that the motility of lichen cyanobionts could be achieved by contraction-relaxation episodes of the cytoskeleton induced by fungal lectin act as a chemoattractant.

  3. The exposed N-terminal tail of the D1 subunit is required for rapid D1 degradation during Photosystem II repair in Synechocystis sp

    Czech Academy of Sciences Publication Activity Database

    Komenda, Josef; Tichý, Martin; Prášil, Ondřej; Knoppová, Jana; Kuviková, Stanislava; de Vries, R.; Nixon, P. J.

    2007-01-01

    Roč. 19, - (2007), s. 2839-2854 ISSN 1040-4651 R&D Projects: GA MŠk LN00A141; GA ČR GA203/04/0800; GA ČR GA206/06/0322 Institutional research plan: CEZ:AV0Z50200510 Keywords : photosystem II * cyanobacterium * synechocystis sp. pcc 6803 Subject RIV: EE - Microbiology, Virology Impact factor: 9.653, year: 2007

  4. The susceptibility of five African Anopheles species to Anabaena PCC 7120 expressing Bacillus thuringiensis subsp. israelensis mosquitocidal cry genes

    Directory of Open Access Journals (Sweden)

    Ketseoglou Irene

    2012-10-01

    Full Text Available Abstract Background Malaria, one of the leading causes of death in Africa, is transmitted by the bite of an infected female Anopheles mosquito. Problems associated with the development of resistance to chemical insecticides and concerns about the non-target effects and persistence of chemical insecticides have prompted the development of environmentally friendly mosquito control agents. The aim of this study was to evaluate the larvicidal activity of a genetically engineered cyanobacterium, Anabaena PCC 7120#11, against five African Anopheles species in laboratory bioassays. Findings There were significant differences in the susceptibility of the anopheline species to PCC 7120#11. The ranking of the larvicidal activity of PCC 7120#11 against species in the An. gambiae complex was: An. merus An. arabiensis An. gambiae An. quadriannulatus, where 50. The LC50 of PCC 7120#11 against the important malaria vectors An. gambiae and An. arabiensis was 12.3 × 105 cells/ml and 8.10 × 105 cells/ml, respectively. PCC 7120#11 was not effective against An. funestus, with less than 50% mortality obtained at concentrations as high as 3.20 × 107 cells/ml. Conclusions PCC 7120#11 exhibited good larvicidal activity against larvae of the An. gambiae complex, but relatively weak larvicidal activity against An. funestus. The study has highlighted the importance of evaluating a novel mosquitocidal agent against a range of malaria vectors so as to obtain a clear understanding of the agent’s spectrum of activity and potential as a vector control agent.

  5. Crystallization and preliminary X-ray crystallographic studies of O-methyltransferase from Anabaena PCC 7120

    International Nuclear Information System (INIS)

    Li, Guoming; Tang, Zhenting; Meng, Geng; Dai, Kesheng; Zhao, Jindong; Zheng, Xiaofeng

    2009-01-01

    The O-methyltransferase (OMT) from the Anabaena PCC 7120 has been overexpressed in a soluble form in E. coli, purified and crystallized. The crystals belonged to space group C222 1 and diffracted to 2.4 Å resolution. O-Methyltransferase (OMT) is a ubiquitous enzyme that exists in bacteria, plants and humans and catalyzes a methyl-transfer reaction using S-adenosyl-l-methionine as a methyl donor and a wide range of phenolics as acceptors. To investigate the structure and function of OMTs, omt from Anabaena PCC 7120 was cloned into expression vector pET21a and expressed in a soluble form in Escherichia coli strain BL21 (DE3). The recombinant OMT protein was purified to homogeneity using a two-step strategy. Crystals of OMT that diffracted to a resolution of 2.4 Å were obtained using the hanging-drop vapour-diffusion method. The crystals belonged to space group C222 1 , with unit-cell parameters a = 131.620, b = 227.994, c = 150.777 Å, α = β = γ = 90°. There are eight molecules per asymmetric unit

  6. Purification, crystallization and preliminary X-ray diffraction of fluorescence recovery protein from Synechocystis PCC 6803

    International Nuclear Information System (INIS)

    Liu, Ting; Shuai, Yingli; Zhou, Honggang

    2011-01-01

    Fluorescence recovery protein from Synechocystis PCC 6803 plays a key role in the orange carotenoid protein-related photoprotective mechanism in cyanobacteria. The full-length form and a truncated form were overexpressed, purified and crystallized, and diffraction was observed to 2.75 Å resolution. Fluorescence recovery protein (FRP), which is encoded by the slr1964 gene in Synechocystis PCC 6803, plays a key role in the orange carotenoid protein-related photoprotective mechanism in cyanobacteria. As the crystal structure of FRP may provide information about the biological functions and mechanism of action of the protein, recombinant full-length FRP and a truncated form were overexpressed, purified and crystallized at 291 K using ethylene imine polymer as the precipitant. An FRP data set was collected to a resolution of 2.75 Å at low temperature (100 K). The crystal belonged to space group P4 1 2 1 2, with unit-cell parameters a = b = 61.9, c = 160.7 Å, α = β = γ = 90°. Assuming that the asymmetric unit contains three molecules, the Matthews coefficient was calculated to be 2.1 Å 3 Da −1

  7. Toxicity of nickel and silver to Nostoc muscorum: interaction with ascorbic acid, glutathione, and sulfur-containing amino acids.

    Science.gov (United States)

    Rai, L C; Raizada, M

    1987-08-01

    Exposure of Nostoc muscorum to different concentrations of Ni and Ag brought about reduction in growth, carbon fixation, heterocyst production, and nitrogenase activity and increase in the loss of ions (K+, Na+). In an attempt to ameliorate the toxicity of test metals by ascorbic acid, glutathione, and sulfur-containing amino acids (L-cysteine and L-methionine), it was found that the level of protection by ascorbic acid and glutathione was more for Ag than Ni. However, metal-induced inhibition of growth and carbon fixation was equally ameliorated by methionine. But the level of protection by cysteine was quite different, i.e., 27% for Ni and 22% for Ag. Protection of metal toxicity in N. muscorum by amino acids lends further support to self-detoxifying ability of cyanobacteria because they are known to synthesize all essential amino acids.

  8. Photoinduced reduction of the medial FeS center in the hydrogenase small subunit HupS from Nostoc punctiforme.

    Science.gov (United States)

    Raleiras, Patrícia; Hammarström, Leif; Lindblad, Peter; Styring, Stenbjörn; Magnuson, Ann

    2015-07-01

    The small subunit from the NiFe uptake hydrogenase, HupSL, in the cyanobacterium Nostoc punctiforme ATCC 29133, has been isolated in the absence of the large subunit (P. Raleiras, P. Kellers, P. Lindblad, S. Styring, A. Magnuson, J. Biol. Chem. 288 (2013) 18,345-18,352). Here, we have used flash photolysis to reduce the iron-sulfur clusters in the isolated small subunit, HupS. We used ascorbate as electron donor to the photogenerated excited state of Ru(II)-trisbipyridine (Ru(bpy)3), to generate Ru(I)(bpy)3 as reducing agent. Our results show that the isolated small subunit can be reduced by the Ru(I)(bpy)3 generated through flash photolysis. Copyright © 2015 Elsevier Inc. All rights reserved.

  9. Effects of light intensity and quality on phycobiliprotein accumulation in the cyanobacterium Nostoc sphaeroides Kützing.

    Science.gov (United States)

    Ma, Rui; Lu, Fan; Bi, Yonghong; Hu, Zhengyu

    2015-08-01

    To assess the effects of light intensity and quality on the growth and phycobiliproteins (PBP) accumulation in Nostoc sphaeroides Kützing (N. sphaeroides). Dry weights, dry matter, protein, chlorophyll and PBP contents were higher under 90 μmol m(-2) s(-1) than under other intensities (both higher and lower). Phycocyanin and allophycocyanin increased with light intensity while phycoerythrin decreased. Fresh weights, protein and PBP contents increased at the highest rates under blue light. Red light resulted in higher values of dry matter, phycocyanin and chlorophyll a. White light at 90 μmol m(-2) s(-1) or blue light 30 μmol m(-2) s(-1) were optimal for the growth and phycobiliprotein accumulation in N. sphaeroides.

  10. A Proposal on the Restoration of Nostoc flagelliforme for Sustainable Improvement in the Ecology of Arid Steppes in China

    Directory of Open Access Journals (Sweden)

    Xiang Gao

    2016-06-01

    Full Text Available Nostoc flagelliforme, a filamentous nitrogen-fixing cyanobacterium, is widely distributed in arid steppes of the west and northwestern parts of China. However, as a food delicacy this species has been overexploited from 1970 to 2000. Moreover, overgrazing, land reclamation and the removal of medicinal herbs have caused severely reduced vegetation coverage there. In this communication, a badly damaged but slowly rehabilitating N. flagelliforme-inhibiting steppe is described, and the rehabilitation of desertified steppes by the renewed growth of N. flagelliforme is proposed. The restoration of this dominant nitrogen supplier would be an ecologically sustainable solution for supplementing current measures already taken in the desertified regions. In addition, a goal of 50%–60% vegetation coverage is proposed by the N. flagelliforme restoration.

  11. Wet season cyanobacterial N enrichment highly correlated with species richness and Nostoc in the northern Australian savannah

    Directory of Open Access Journals (Sweden)

    W. Williams

    2018-04-01

    Full Text Available The Boodjamulla National Park research station is situated in the north-western Queensland dry savannah, where the climate is dominated by summer monsoons and virtually dry winters. Under shrub canopies and in between the tussock grasses cyanobacterial crusts almost entirely cover the flood plain soil surfaces. Seasonality drives N fixation, and in the savannah this has a large impact on both plant and soil function. Many cyanobacteria fix dinitrogen that is liberated into the soil in both inorganic and organic N forms. We examined cyanobacterial species richness and bioavailable N spanning 7 months of a typical wet season. Over the wet season cyanobacterial richness ranged from 6 to 19 species. N-fixing Scytonema accounted for seasonal averages between 51 and 93 % of the biocrust. Cyanobacterial richness was highly correlated with N fixation and bioavailable N in 0–1 cm. Key N-fixing species such as Nostoc, Symploca and Gloeocapsa significantly enriched soil N although Nostoc was the most influential. Total seasonal N fixation by cyanobacteria demonstrated the variability in productivity according to the number of wet days as well as the follow-on days where the soil retained adequate moisture. Based on total active days per month we estimated that N soil enrichment via cyanobacteria would be  ∼  5.2 kg ha−1 annually which is comparable to global averages. This is a substantial contribution to the nutrient-deficient savannah soils that are almost entirely reliant on the wet season for microbial turnover of organic matter. Such well-defined seasonal trends and synchronisation in cyanobacterial species richness, N fixation, bioavailable N and C fixation (Büdel et al., 2018 provide important contributions to multifunctional microprocesses and soil fertility.

  12. Wet season cyanobacterial N enrichment highly correlated with species richness and Nostoc in the northern Australian savannah

    Science.gov (United States)

    Williams, Wendy; Büdel, Burkhard; Williams, Stephen

    2018-04-01

    The Boodjamulla National Park research station is situated in the north-western Queensland dry savannah, where the climate is dominated by summer monsoons and virtually dry winters. Under shrub canopies and in between the tussock grasses cyanobacterial crusts almost entirely cover the flood plain soil surfaces. Seasonality drives N fixation, and in the savannah this has a large impact on both plant and soil function. Many cyanobacteria fix dinitrogen that is liberated into the soil in both inorganic and organic N forms. We examined cyanobacterial species richness and bioavailable N spanning 7 months of a typical wet season. Over the wet season cyanobacterial richness ranged from 6 to 19 species. N-fixing Scytonema accounted for seasonal averages between 51 and 93 % of the biocrust. Cyanobacterial richness was highly correlated with N fixation and bioavailable N in 0-1 cm. Key N-fixing species such as Nostoc, Symploca and Gloeocapsa significantly enriched soil N although Nostoc was the most influential. Total seasonal N fixation by cyanobacteria demonstrated the variability in productivity according to the number of wet days as well as the follow-on days where the soil retained adequate moisture. Based on total active days per month we estimated that N soil enrichment via cyanobacteria would be ˜ 5.2 kg ha-1 annually which is comparable to global averages. This is a substantial contribution to the nutrient-deficient savannah soils that are almost entirely reliant on the wet season for microbial turnover of organic matter. Such well-defined seasonal trends and synchronisation in cyanobacterial species richness, N fixation, bioavailable N and C fixation (Büdel et al., 2018) provide important contributions to multifunctional microprocesses and soil fertility.

  13. Production of precipitated calcium carbonate from industrial byproduct slags; Saostetun kalsiumkarbonaatin tuotanto karbonaattivapaista kuonatuotteista (SLAG2PCC)

    Energy Technology Data Exchange (ETDEWEB)

    Zevenhoven, R. [Aabo Akademi, Turku (Finland). Heat Engineering Lab.; Teir, S.; Eloneva, S.; Savolahti, J. [Helsinki Univ. of Technology, Espoo (Finland). Energy Technology and Environmental Protection

    2006-12-19

    Production of precipitate calcium carbonate from industrial by- product slags-project, 'SLAG2PCC', is a spin-off from ClimBus technology programme CO{sub 2} Nordic Plus-project, financed by the Finnish Technology Agency Tekes and the Finnish Recovery Boiler Committee. 'SLAG2PCC'-project is financed by Tekes, Ruukki Productions, UPM Kymmene and Waertsilae Finland. The possibility to produce precipitated calcium carbonate, PCC, from carbonate free industrial by-products (slags), combined with binding of carbon dioxide for climate change mitigation is studied in this project. The suitability of a process found from the literature, in which calcium used for carbonation is dissolved from calcium silicates using acetic acid as a solvent, is investigated for the carbonation of slags from the steel industry. During the calcium extraction experiments performed in the CO2 Nordic Plus - project it was found out that calcium is rapidly extracted from blast furnace and basic oxygen furnace slags. Atmospheric carbonation of the solution containing the dissolved slag and acetic acid directly has not succeeded yet due to low pH of the solution. Addition of NaOH, to increase of the solution pH, resulted in calcium carbonate precipitate in atmospheric pressure. The future goal of the project is to optimize process conditions so that the formed calcium carbonate is suitable for use as PCC. (orig.)

  14. Techniques for induction of premature chromosome condensation (PCC) by Calyculin-A and micronucleus assay for biodosimetry in Vietnam

    International Nuclear Information System (INIS)

    Pham Ngoc Duy; Tran Que; Hoang Hung Tien; Bui Thi Kim Luyen; Nguyen Thi Kim Anh; Ha Thi Ngoc Lien

    2014-01-01

    The International Atomic Energy Agency (IAEA) and World Health Organization are interested in biological dosimetry method for radiation emergency medicine currently. Some cytogenetic techniques such as premature chromosome condensation (PCC) induced by Calyculin-A and micronucleus (MN) assay are necessary to develop biodosimetry in Vietnam. In this study, we optimized the condition for MN assay with 6 µg/ml Cytochalasin-B concentration and 72.5 hours for peripheral lymphocyte blood culture. The optimization for PCC method is 50 nM Calyculin-A concentration for 45 minutes peripheral lymphocyte blood treatment. For samples exposed to 3.0 Gy gamma 60 Co (dose rate 0.0916 Gy/s), the frequency of MN is 19.02 ± 0.38%, NBP is 1.95 ± 0.28%, dicentric and ring is 41.43 ± 8.12% and frag and min is 63.33 ± 5.16%. For samples exposed to 6.0 Gy gamma 60 Co (dose rate 0.0916 Gy/s), the frequency of ring-PCC is 17.73 ± 2.46%, extra unite is 218.91± 7.58%, dicentric is 83.81 ± 1.09%, ring is 10.75 ± 1.74%, fragment and minute is 193.17 ± 13.10%. MN and ring-PCC are specific marker applying for biodosimetry. (author)

  15. Engineering Synechocystis PCC6803 for hydrogen production: influence on the tolerance to oxidative and sugar stresses.

    Directory of Open Access Journals (Sweden)

    Marcia Ortega-Ramos

    Full Text Available In the prospect of engineering cyanobacteria for the biological photoproduction of hydrogen, we have studied the hydrogen production machine in the model unicellular strain Synechocystis PCC6803 through gene deletion, and overexpression (constitutive or controlled by the growth temperature. We demonstrate that the hydrogenase-encoding hoxEFUYH operon is dispensable to standard photoautotrophic growth in absence of stress, and it operates in cell defense against oxidative (H₂O₂ and sugar (glucose and glycerol stresses. Furthermore, we showed that the simultaneous over-production of the proteins HoxEFUYH and HypABCDE (assembly of hydrogenase, combined to an increase in nickel availability, led to an approximately 20-fold increase in the level of active hydrogenase. These novel results and mutants have major implications for those interested in hydrogenase, hydrogen production and redox metabolism, and their connections with environmental conditions.

  16. Structural Insights into l-Tryptophan Dehydrogenase from a Photoautotrophic Cyanobacterium, Nostoc punctiforme.

    Science.gov (United States)

    Wakamatsu, Taisuke; Sakuraba, Haruhiko; Kitamura, Megumi; Hakumai, Yuichi; Fukui, Kenji; Ohnishi, Kouhei; Ashiuchi, Makoto; Ohshima, Toshihisa

    2017-01-15

    l-Tryptophan dehydrogenase from Nostoc punctiforme NIES-2108 (NpTrpDH), despite exhibiting high amino acid sequence identity (>30%)/homology (>50%) with NAD(P) + -dependent l-Glu/l-Leu/l-Phe/l-Val dehydrogenases, exclusively catalyzes reversible oxidative deamination of l-Trp to 3-indolepyruvate in the presence of NAD + Here, we determined the crystal structure of the apo form of NpTrpDH. The structure of the NpTrpDH monomer, which exhibited high similarity to that of l-Glu/l-Leu/l-Phe dehydrogenases, consisted of a substrate-binding domain (domain I, residues 3 to 133 and 328 to 343) and an NAD + /NADH-binding domain (domain II, residues 142 to 327) separated by a deep cleft. The apo-NpTrpDH existed in an open conformation, where domains I and II were apart from each other. The subunits dimerized themselves mainly through interactions between amino acid residues around the β-1 strand of each subunit, as was observed in the case of l-Phe dehydrogenase. The binding site for the substrate l-Trp was predicted by a molecular docking simulation and validated by site-directed mutagenesis. Several hydrophobic residues, which were located in the active site of NpTrpDH and possibly interacted with the side chain of the substrate l-Trp, were arranged similarly to that found in l-Leu/l-Phe dehydrogenases but fairly different from that of an l-Glu dehydrogenase. Our crystal structure revealed that Met-40, Ala-69, Ile-74, Ile-110, Leu-288, Ile-289, and Tyr-292 formed a hydrophobic cluster around the active site. The results of the site-directed mutagenesis experiments suggested that the hydrophobic cluster plays critical roles in protein folding, l-Trp recognition, and catalysis. Our results provide critical information for further characterization and engineering of this enzyme. In this study, we determined the three-dimensional structure of l-Trp dehydrogenase, analyzed its various site-directed substitution mutants at residues located in the active site, and obtained the

  17. A Genetic and Chemical Perspective on Symbiotic Recruitment of Cyanobacteria of the Genus Nostoc into the Host Plant Blasia pusilla L.

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    Anton Liaimer

    2016-11-01

    Full Text Available Liverwort Blasia pusilla L. recruits soil nitrogen-fixing cyanobacteria of genus Nostoc as symbiotic partners. In this work we compared Nostoc community composition inside the plants and in the soil around them from two distant locations in Northern Norway. STRR fingerprinting and 16S rDNA phylogeny reconstruction showed a remarkable local diversity among isolates assigned to several Nostoc clades. An extensive web of negative allelopathic interactions was recorded at an agricultural site, but not at the undisturbed natural site. The cell extracts of the cyanobacteria did not show antimicrobial activities, but four isolates were shown to be cytotoxic to human cells. The secondary metabolite profiles of the isolates were mapped by MALDI-TOF MS, and the most prominent ions were further analysed by Q-TOF for MS/MS aided identification. Symbiotic isolates produced a great variety of small peptide-like substances, most of which lack any record in the databases. Among identified compounds we found microcystin and nodularin variants toxic to eukaryotic cells. Microcystin producing chemotypes were dominating as symbiotic recruits but not in the free-living community. In addition, we were able to identify several novel aeruginosins and banyaside-like compounds, as well as nostocyclopeptides and nosperin.

  18. A Genetic and Chemical Perspective on Symbiotic Recruitment of Cyanobacteria of the Genus Nostoc into the Host Plant Blasia pusilla L.

    Science.gov (United States)

    Liaimer, Anton; Jensen, John B.; Dittmann, Elke

    2016-01-01

    Liverwort Blasia pusilla L. recruits soil nitrogen-fixing cyanobacteria of genus Nostoc as symbiotic partners. In this work we compared Nostoc community composition inside the plants and in the soil around them from two distant locations in Northern Norway. STRR fingerprinting and 16S rDNA phylogeny reconstruction showed a remarkable local diversity among isolates assigned to several Nostoc clades. An extensive web of negative allelopathic interactions was recorded at an agricultural site, but not at the undisturbed natural site. The cell extracts of the cyanobacteria did not show antimicrobial activities, but four isolates were shown to be cytotoxic to human cells. The secondary metabolite profiles of the isolates were mapped by MALDI-TOF MS, and the most prominent ions were further analyzed by Q-TOF for MS/MS aided identification. Symbiotic isolates produced a great variety of small peptide-like substances, most of which lack any record in the databases. Among identified compounds we found microcystin and nodularin variants toxic to eukaryotic cells. Microcystin producing chemotypes were dominating as symbiotic recruits but not in the free-living community. In addition, we were able to identify several novel aeruginosins and banyaside-like compounds, as well as nostocyclopeptides and nosperin. PMID:27847500

  19. ExaSP2

    Energy Technology Data Exchange (ETDEWEB)

    2017-09-08

    ExaSP2 is a reference implementation of typical linear algebra algorithms and workloads for a quantum molecular dynamics (QMD) electronic structure code. The algorithm is based on a recursive second-order Fermi-Operator expansion method (SP2) and is tailored for density functional based tight-binding calculations of material systems.

  20. A homozygous recA mutant of Synechocystis PCC6803: construction strategy and characteristics eliciting a novel RecA independent UVC resistance in dark.

    Science.gov (United States)

    Minda, Renu; Ramchandani, Jyoti; Joshi, Vasudha P; Bhattacharjee, Swapan Kumar

    2005-12-01

    We report here the construction of a homozygous recA460::cam insertion mutant of Synechocystis sp. PCC 6803 that may be useful for plant molecular genetics by providing a plant like host free of interference from homologous recombination. The homozygous recA460::cam mutant is highly sensitive to UVC under both photoreactivating and non-photoreactivating conditions compared to the wild type (WT). The liquid culture of the mutant growing in approximately 800 lx accumulates nonviable cells to the tune of 86% as estimated by colony counts on plates incubated at the same temperature and light intensity. The generation time of recA mutant in standard light intensity (2,500 lx) increases to 50 h compared to 28 h in lower light intensity (approximately 800 lx) that was used for selection, thus explaining the earlier failures to obtain a homozygous recA mutant. The WT, in contrast, grows at faster rate (23 h generation time) in standard light intensity compared to that at approximately 800 lx (26 h). The Synechocystis RecA protein supports homologous recombination during conjugation in recA (-) mutant of Escherichia coli, but not the SOS response as measured by UV sensitivity. It is suggested that using this homozygous recA460::cam mutant, investigations can now be extended to dissect the network of DNA repair pathways involved in housekeeping activities that may be more active in cyanobacteria than in heterotrophs. Using this mutant for the first time we provide a genetic evidence of a mechanism independent of RecA that causes enhanced UVC resistance on light to dark transition.

  1. Involvement of Potassium Transport Systems in the Response of Synechocystis PCC 6803 Cyanobacteria to External pH Change, High-Intensity Light Stress and Heavy Metal Stress.

    Science.gov (United States)

    Checchetto, Vanessa; Segalla, Anna; Sato, Yuki; Bergantino, Elisabetta; Szabo, Ildiko; Uozumi, Nobuyuki

    2016-04-01

    The unicellular photosynthetic cyanobacterium, able to survive in varying environments, is the only prokaryote that directly converts solar energy and CO2 into organic material and is thus relevant for primary production in many ecosystems. To maintain the intracellular and intrathylakoid ion homeostasis upon different environmental challenges, the concentration of potassium as a major intracellular cation has to be optimized by various K(+)uptake-mediated transport systems. We reveal here the specific and concerted physiological function of three K(+)transporters of the plasma and thylakoid membranes, namely of SynK (K(+)channel), KtrB (Ktr/Trk/HKT) and KdpA (Kdp) in Synechocystis sp. strain PCC 6803, under specific stress conditions. The behavior of the wild type, single, double and triple mutants was compared, revealing that only Synk contributes to heavy metal-induced stress, while only Ktr/Kdp is involved in osmotic and salt stress adaptation. With regards to pH shifts in the external medium, the Kdp/Ktr uptake systems play an important role in the adaptation to acidic pH. Ktr, by affecting the CO2 concentration mechanism via its action on the bicarbonate transporter SbtA, might also be responsible for the observed effects concerning high-light stress and calcification. In the case of illumination with high-intensity light, a synergistic action of Kdr/Ktp and SynK is required in order to avoid oxidative stress and ensure cell viability. In summary, this study dissects, using growth tests, measurement of photosynthetic activity and analysis of ultrastructure, the physiological role of three K(+)transporters in adaptation of the cyanobacteria to various environmental changes. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  2. Lack of Methylated Hopanoids Renders the Cyanobacterium Nostoc punctiforme Sensitive to Osmotic and pH Stress.

    Science.gov (United States)

    Garby, Tamsyn J; Matys, Emily D; Ongley, Sarah E; Salih, Anya; Larkum, Anthony W D; Walter, Malcolm R; Summons, Roger E; Neilan, Brett A

    2017-07-01

    To investigate the function of 2-methylhopanoids in modern cyanobacteria, the hpnP gene coding for the radical S -adenosyl methionine (SAM) methylase protein that acts on the C-2 position of hopanoids was deleted from the filamentous cyanobacterium Nostoc punctiforme ATCC 29133S. The resulting Δ hpnP mutant lacked all 2-methylhopanoids but was found to produce much higher levels of two bacteriohopanepentol isomers than the wild type. Growth rates of the Δ hpnP mutant cultures were not significantly different from those of the wild type under standard growth conditions. Akinete formation was also not impeded by the absence of 2-methylhopanoids. The relative abundances of the different hopanoid structures in akinete-dominated cultures of the wild-type and Δ hpnP mutant strains were similar to those of vegetative cell-dominated cultures. However, the Δ hpnP mutant was found to have decreased growth rates under both pH and osmotic stress, confirming a role for 2-methylhopanoids in stress tolerance. Evidence of elevated photosystem II yield and NAD(P)H-dependent oxidoreductase activity in the Δ hpnP mutant under stress conditions, compared to the wild type, suggested that the absence of 2-methylhopanoids increases cellular metabolic rates under stress conditions. IMPORTANCE As the first group of organisms to develop oxygenic photosynthesis, Cyanobacteria are central to the evolutionary history of life on Earth and the subsequent oxygenation of the atmosphere. To investigate the origin of cyanobacteria and the emergence of oxygenic photosynthesis, geobiologists use biomarkers, the remnants of lipids produced by different organisms that are found in geologic sediments. 2-Methylhopanes have been considered indicative of cyanobacteria in some environmental settings, with the parent lipids 2-methylhopanoids being present in many contemporary cyanobacteria. We have created a Nostoc punctiforme Δ hpnP mutant strain that does not produce 2-methylhopanoids to assess the

  3. EasyPCC: Benchmark Datasets and Tools for High-Throughput Measurement of the Plant Canopy Coverage Ratio under Field Conditions

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    Wei Guo

    2017-04-01

    Full Text Available Understanding interactions of genotype, environment, and management under field conditions is vital for selecting new cultivars and farming systems. Image analysis is considered a robust technique in high-throughput phenotyping with non-destructive sampling. However, analysis of digital field-derived images remains challenging because of the variety of light intensities, growth environments, and developmental stages. The plant canopy coverage (PCC ratio is an important index of crop growth and development. Here, we present a tool, EasyPCC, for effective and accurate evaluation of the ground coverage ratio from a large number of images under variable field conditions. The core algorithm of EasyPCC is based on a pixel-based segmentation method using a decision-tree-based segmentation model (DTSM. EasyPCC was developed under the MATLAB® and R languages; thus, it could be implemented in high-performance computing to handle large numbers of images following just a single model training process. This study used an experimental set of images from a paddy field to demonstrate EasyPCC, and to show the accuracy improvement possible by adjusting key points (e.g., outlier deletion and model retraining. The accuracy (R2 = 0.99 of the calculated coverage ratio was validated against a corresponding benchmark dataset. The EasyPCC source code is released under GPL license with benchmark datasets of several different crop types for algorithm development and for evaluating ground coverage ratios.

  4. The effect of coatings and coating weight by two types of PCC on barrier and optical properties and roughness of paper

    Directory of Open Access Journals (Sweden)

    rouzbeh asadi khansari

    2017-08-01

    Full Text Available The objective of this work is to investigate the use of PCC, and the impact of its coating weight on paper coating. In this study, two base papers from Mazandaran Wood and Paper Industries (APC and NS, and two coating compositions with the solid content of 25% containing PCC filler (100 parts, PVA binder (14 parts and dispersant (1 part were used. The first composition included PCC B102 for opacity increment, and the second one PCC 9020 for the improvement of brightness. Two rod RDS14 and RDS30 were used for different coating weights. After coating, the treated samples were dried in room conditions at air temperature of 25◦C and relative humidity of 54%. Physical and optical properties of control and treated samples such as air resistance, thickness, Cobb60, brightness, yellowness, opacity and roughness were determined. In comparison to the control group, all the treated samples showed improvement in brightness, opacity, yellowness and air resistance. By the two different formulations and two rods, paper roughness was increased, and the increment of water absorption was due to capillary development in coating texture. The analysis of variances showed that the usage of PCC 9020 had considerable effect on roughness of papers. In NS papers, change of PCC caused significant difference in brightness and roughness, but in APC papers did not. The change of coating rod in APC papers had significant effect on water absorption, brightness and opacity but did not show in NS.

  5. Using a Microfluidic Gradient Generator to Characterize BG-11 Medium for the Growth of Cyanobacteria Synechococcus elongatus PCC7942

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    Chih-Chun Yang

    2015-11-01

    Full Text Available The photosynthetic cyanobacterium Synechococcus elongatus PCC7942 has recently gained great attention for its ability to directly convert CO2 into renewable chemicals upon genetic engineering. Thus, it is of great interest to increase the growth speed and lower the medium requirement for cultivating this cyanobacterium. The cultivation medium of Synechococcus elongatus PCC7942 has been developed, which consists of many inorganic and metal ingredients with a specific composition, known as the BG-11 medium. In this work, we analyzed the concentration effect of each ingredient and identified the absolutely essential components in BG-11 medium for cyanobacteria growth using the concentration gradient generator chip (CGGC fabricated by MEMS technology. As shown by our results, removal of the individual component sodium nitrate, potassium phosphate, or magnesium sulfate from the BG-11 medium led to severe growth inhibition of Synechococcus elongatus PCC7942. Contrary to our expectation, increasing concentration of the crucial ingredients showed either insignificant or negative impact on cell growth. Overall, standard growth could be achieved without supplementation of ethylenediaminetetraacetic acid (EDTA disodium, sodium carbonate, or sodium citrate to the culture medium. Further improvement of the CGGC-based microfluidic system based on this preliminary study may broaden its application range to analyze more complicated correlations.

  6. Function and Regulation of Ferredoxins in the Cyanobacterium, Synechocystis PCC6803: Recent Advances.

    Science.gov (United States)

    Cassier-Chauvat, Corinne; Chauvat, Franck

    2014-11-07

    Ferredoxins (Fed), occurring in most organisms, are small proteins that use their iron-sulfur cluster to distribute electrons to various metabolic pathways, likely including hydrogen production. Here, we summarize the current knowledge on ferredoxins in cyanobacteria, the prokaryotes regarded as important producers of the oxygenic atmosphere and biomass for the food chain, as well as promising cell factories for biofuel production. Most studies of ferredoxins were performed in the model strain, Synechocystis PCC6803, which possesses nine highly-conserved ferredoxins encoded by monocistronic or operonic genes, some of which are localized in conserved genome regions. Fed1, encoded by a light-inducible gene, is a highly abundant protein essential to photosynthesis. Fed2-Fed9, encoded by genes differently regulated by trophic conditions, are low-abundant proteins that play prominent roles in the tolerance to environmental stresses. Concerning the selectivity/redundancy of ferredoxin, we report that Fed1, Fed7 and Fed9 belong to ferredoxin-glutaredoxin-thioredoxin crosstalk pathways operating in the protection against oxidative and metal stresses. Furthermore, Fed7 specifically interacts with a DnaJ-like protein, an interaction that has been conserved in photosynthetic eukaryotes in the form of a composite protein comprising DnaJ- and Fed7-like domains. Fed9 specifically interacts with the Flv3 flavodiiron protein acting in the photoreduction of O2 to H2O.

  7. Function and Regulation of Ferredoxins in the Cyanobacterium, Synechocystis PCC6803: Recent Advances

    Directory of Open Access Journals (Sweden)

    Corinne Cassier-Chauvat

    2014-11-01

    Full Text Available Ferredoxins (Fed, occurring in most organisms, are small proteins that use their iron-sulfur cluster to distribute electrons to various metabolic pathways, likely including hydrogen production. Here, we summarize the current knowledge on ferredoxins in cyanobacteria, the prokaryotes regarded as important producers of the oxygenic atmosphere and biomass for the food chain, as well as promising cell factories for biofuel production. Most studies of ferredoxins were performed in the model strain, Synechocystis PCC6803, which possesses nine highly-conserved ferredoxins encoded by monocistronic or operonic genes, some of which are localized in conserved genome regions. Fed1, encoded by a light-inducible gene, is a highly abundant protein essential to photosynthesis. Fed2-Fed9, encoded by genes differently regulated by trophic conditions, are low-abundant proteins that play prominent roles in the tolerance to environmental stresses. Concerning the selectivity/redundancy of ferredoxin, we report that Fed1, Fed7 and Fed9 belong to ferredoxin-glutaredoxin-thioredoxin crosstalk pathways operating in the protection against oxidative and metal stresses. Furthermore, Fed7 specifically interacts with a DnaJ-like protein, an interaction that has been conserved in photosynthetic eukaryotes in the form of a composite protein comprising DnaJ- and Fed7-like domains. Fed9 specifically interacts with the Flv3 flavodiiron protein acting in the photoreduction of O2 to H2O.

  8. Advances in the Function and Regulation of Hydrogenase in the Cyanobacterium Synechocystis PCC6803

    Science.gov (United States)

    Cassier-Chauvat, Corinne; Veaudor, Théo; Chauvat, Franck

    2014-01-01

    In order to use cyanobacteria for the biological production of hydrogen, it is important to thoroughly study the function and the regulation of the hydrogen-production machine in order to better understand its role in the global cell metabolism and identify bottlenecks limiting H2 production. Most of the recent advances in our understanding of the bidirectional [Ni-Fe] hydrogenase (Hox) came from investigations performed in the widely-used model cyanobacterium Synechocystis PCC6803 where Hox is the sole enzyme capable of combining electrons with protons to produce H2 under specific conditions. Recent findings suggested that the Hox enzyme can receive electrons from not only NAD(P)H as usually shown, but also, or even preferentially, from ferredoxin. Furthermore, plasmid-encoded functions and glutathionylation (the formation of a mixed-disulfide between the cysteines residues of a protein and the cysteine residue of glutathione) are proposed as possible new players in the function and regulation of hydrogen production. PMID:25365180

  9. Isobutanol production in Synechocystis PCC 6803 using heterologous and endogenous alcohol dehydrogenases

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    Rui Miao

    2017-12-01

    Full Text Available Isobutanol is a flammable compound that can be used as a biofuel due to its high energy density and suitable physical and chemical properties. In this study, we examined the capacity of engineered strains of Synechocystis PCC 6803 containing the α-ketoisovalerate decarboxylase from Lactococcus lactis and different heterologous and endogenous alcohol dehydrogenases (ADH for isobutanol production. A strain expressing an introduced kivd without any additional copy of ADH produced 3 mg L−1 OD750−1 isobutanol in 6 days. After the cultures were supplemented with external addition of isobutyraldehyde, the substrate for ADH, 60.8 mg L−1 isobutanol was produced after 24 h when OD750 was 0.8. The in vivo activities of four different ADHs, two heterologous and two putative endogenous in Synechocystis, were examined and the Synechocystis endogenous ADH encoded by slr1192 showed the highest efficiency for isobutanol production. Furthermore, the strain overexpressing the isobutanol pathway on a self-replicating vector with the strong Ptrc promoter showed significantly higher gene expression and isobutanol production compared to the corresponding strains expressing the same operon introduced on the genome. Hence, this study demonstrates that Synechocystis endogenous AHDs have a high capacity for isobutanol production, and identifies kivd encoded α-ketoisovalerate decarboxylase as one of the likely bottlenecks for further isobutanol production.

  10. PfsR is a key regulator of iron homeostasis in Synechocystis PCC 6803.

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    Dan Cheng

    Full Text Available Iron is an essential cofactor in numerous cellular processes. The iron deficiency in the oceans affects the primary productivity of phytoplankton including cyanobacteria. In this study, we examined the function of PfsR, a TetR family transcriptional regulator, in iron homeostasis of the cyanobacterium Synechocystis PCC 6803. Compared with the wild type, the pfsR deletion mutant displayed stronger tolerance to iron limitation and accumulated significantly more chlorophyll a, carotenoid, and phycocyanin under iron-limiting conditions. The mutant also maintained more photosystem I and photosystem II complexes than the wild type after iron deprivation. In addition, the activities of photosystem I and photosystem II were much higher in pfsR deletion mutant than in wild-type cells under iron-limiting conditions. The transcripts of pfsR were enhanced by iron limitation and inactivation of the gene affected pronouncedly expression of fut genes (encoding a ferric iron transporter, feoB (encoding a ferrous iron transporter, bfr genes (encoding bacterioferritins, ho genes (encoding heme oxygenases, isiA (encoding a chlorophyll-binding protein, and furA (encoding a ferric uptake regulator. The iron quota in pfsR deletion mutant cells was higher than in wild-type cells both before and after exposure to iron limitation. Electrophoretic mobility shift assays showed that PfsR bound to its own promoter and thereby auto-regulated its own expression. These data suggest that PfsR is a critical regulator of iron homeostasis.

  11. Effects of radio frequency and high pressure steam sterilisation on the colour and flavour of prepared Nostoc sphaeroides.

    Science.gov (United States)

    Xu, Jicheng; Zhang, Min; An, Yanjun; Roknul, Azam Sm; Adhikari, Benu

    2018-03-01

    Nostoc sphaeroides has been used as a highly effective herbal medicine and dietary supplement for thousands of years. The desired dark green colour of fresh N. sphaeroides is converted into an undesirable dark brown during conventional high pressure (HP) steam sterilisation. Radio frequency (RF) sterilisation technology was used in this study to determine its effectiveness in sterilising N. sphaeroides and to achieve better preservation of natural colour and desirable flavour. Sterilisation was carried out using a 6 kW, 27 MHz RF instrument for 10, 20 and 30 min. The degree of microbial kill and the effects of RF sterilisation on colour and flavour were determined and compared with those obtained from HP steam (121 °C, 30 min) sterilisation. The effects of RF sterilisation on colour and flavour (measured using electronic nose) parameters were significantly lower than that in HP steam sterilisation. The RF sterilisation carried out for 20 min achieved logarithmic reduction of bacterial population and met China's national standard while preserving the colour and flavour better. Results of the present study indicated that application of RF sterilisation would improve the quality of sterilised N. sphaeroides and broaden its application in the food and health food industries. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  12. Evolution of the tRNALeu (UAA) Intron and Congruence of Genetic Markers in Lichen-Symbiotic Nostoc.

    Science.gov (United States)

    Kaasalainen, Ulla; Olsson, Sanna; Rikkinen, Jouko

    2015-01-01

    The group I intron interrupting the tRNALeu UAA gene (trnL) is present in most cyanobacterial genomes as well as in the plastids of many eukaryotic algae and all green plants. In lichen symbiotic Nostoc, the P6b stem-loop of trnL intron always involves one of two different repeat motifs, either Class I or Class II, both with unresolved evolutionary histories. Here we attempt to resolve the complex evolution of the two different trnL P6b region types. Our analysis indicates that the Class II repeat motif most likely appeared first and that independent and unidirectional shifts to the Class I motif have since taken place repeatedly. In addition, we compare our results with those obtained with other genetic markers and find strong evidence of recombination in the 16S rRNA gene, a marker widely used in phylogenetic studies on Bacteria. The congruence of the different genetic markers is successfully evaluated with the recently published software Saguaro, which has not previously been utilized in comparable studies.

  13. Quality evaluation of the edible blue-green alga Nostoc flagelliforme using a chlorophyll fluorescence parameter and several biochemical markers.

    Science.gov (United States)

    Gao, Xiang; Yang, Yiwen; Ai, Yufeng; Luo, Hongyi; Qiu, Baosheng

    2014-01-15

    Nostoc flagelliforme is an edible blue-green alga with herbal and dietary values. Due to the diminishing supply of natural N. flagelliforme and the large investment on the development of its cultivation technology, it is anticipated that artificially cultured N. flagelliforme will soon sustain the market supply. Once this change occurs, the storage-associated quality problem will become the focus of attention for future trade. In this paper, we used a chlorophyll fluorescence parameter, maximum quantum efficiency of Photosystem II (Fv/Fm), and several biomarkers to evaluate the quality of several N. flagelliforme samples. It was found that longer storage times resulted in darker coloured solutions (released pigments) and decreased amounts of chlorophyll a (Chl a) and water-soluble sugars (WSS). Additionally, a higher Fv/Fm value suggests better physiological recovery and quality. In actual application, determination of Fv/Fm would be the first step for evaluating the quality of N. flagelliforme, and the biochemical indexes would serve as good secondary markers. Copyright © 2013 Elsevier Ltd. All rights reserved.

  14. Dynamic localization of HmpF regulates type IV pilus activity and directional motility in the filamentous cyanobacterium Nostoc punctiforme.

    Science.gov (United States)

    Cho, Ye Won; Gonzales, Alfonso; Harwood, Thomas V; Huynh, Jessica; Hwang, Yeji; Park, Jun Sang; Trieu, Anthony Q; Italia, Parth; Pallipuram, Vivek K; Risser, Douglas D

    2017-10-01

    Many cyanobacteria exhibit surface motility powered by type 4 pili (T4P). In the model filamentous cyanobacterium Nostoc punctiforme, the T4P systems are arrayed in static, bipolar rings in each cell. The chemotaxis-like Hmp system is essential for motility and the coordinated polar accumulation of PilA on cells in motile filaments, while the Ptx system controls positive phototaxis. Using transposon mutagenesis, a gene, designated hmpF, was identified as involved in motility. Synteny among filamentous cyanobacteria and the similar expression patterns for hmpF and hmpD imply that HmpF is part of the Hmp system. Deletion of hmpF produced a phenotype distinct from other hmp genes, but indistinguishable from pilB or pilQ. Both an HmpF-GFPuv fusion protein, and PilA, as assessed by in situ immunofluorescence, displayed coordinated, unipolar localization at the leading pole of each cell. Reversals were modulated by changes in light intensity and preceded by the migration of HmpF-GFPuv to the lagging cell poles. These results are consistent with a model where direct interaction between HmpF and the T4P system activates pilus extension, the Hmp system facilitates coordinated polarity of HmpF to establish motility, and the Ptx system modulates HmpF localization to initiate reversals in response to changes in light intensity. © 2017 John Wiley & Sons Ltd.

  15. Gene expression of a two-component regulatory system associated with sunscreen biosynthesis in the cyanobacterium Nostoc punctiforme ATCC 29133.

    Science.gov (United States)

    Janssen, Jacob; Soule, Tanya

    2016-01-01

    Long-wavelength ultraviolet radiation (UVA) can damage cells through photooxidative stress, leading to harmful photosensitized proteins and pigments in cyanobacteria. To mitigate damage, some cyanobacteria secrete the UVA-absorbing pigment scytonemin into their extracellular sheath. Comparative genomic analyses suggest that scytonemin biosynthesis is regulated by the two-component regulatory system (TCRS) proteins encoded by Npun_F1277 and Npun_F1278 in the cyanobacterium Nostoc punctiforme ATCC 29133. To understand the dynamics of these genes, their expression was measured following exposure to UVA, UVB, high visible (VIS) irradiance and oxidative stress for 20, 40 and 60 min. Overall, both genes had statistically similar patterns of expression for all four conditions and were generally upregulated, except for those exposed to UVB by 60 min and for the cells under oxidative stress. The greatest UVA response was an upregulation by 20 min, while the response to UVB was the most dramatic and persisted through 40 min. High VIS irradiance resulted in a modest upregulation, while oxidative stress caused a slight downregulation. Both genes were also found to occur on the same transcript. These results demonstrate that these genes are positively responding to several light-associated conditions, which suggests that this TCRS may regulate more than just scytonemin biosynthesis under UVA stress. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  16. The effects of the exopolysaccharide and growth rate on the morphogenesis of the terrestrial filamentous cyanobacterium Nostoc flagelliforme

    Directory of Open Access Journals (Sweden)

    Lijuan Cui

    2017-09-01

    Full Text Available The terrestrial cyanobacterium Nostoc flagelliforme, which contributes to carbon and nitrogen supplies in arid and semi-arid regions, adopts a filamentous colony form. Owing to its herbal and dietary values, this species has been overexploited. Largely due to the lack of understanding on its morphogenesis, artificial cultivation has not been achieved. Additionally, it may serve as a useful model for recognizing the morphological adaptation of colonial cyanobacteria in terrestrial niches. However, it shows very slow growth in native habitats and is easily disintegrated under laboratory conditions. Thus, a novel experimental system is necessary to explore its morphogenetic mechanism. Liquid-cultured N. flagelliforme has been well developed for exopolysaccharide (EPS production, in which microscopic colonies (micro-colonies are generally formed. In this study, we sought to gain some insight into the morphogenesis of N. flagelliforme by examining the effects of two external factors, the EPS and environmental stress-related growth rate, on the morphological shaping of micro-colonies. Our findings indicate that the EPS matrix could act as a basal barrier, leading to the bending of trichomes during their elongation, while very slow growth is conducive to their straight elongation. These findings will guide future cultivation and application of this cyanobacterium for ecological improvement.

  17. 2-Methyl-3-buten-2-ol (MBO) synthase expression in Nostoc punctiforme leads to over production of phytols.

    Science.gov (United States)

    Gupta, Dinesh; Ip, Tina; Summers, Michael L; Basu, Chhandak

    2015-01-01

    Phytol is a diterpene alcohol of medicinal importance and it also has potential to be used as biofuel. We found over production of phytol in Nostoc punctiforme by expressing a 2-Methyl-3-buten-2-ol (MBO) synthase gene. MBO synthase catalyzes the conversion of dimethylallyl pyrophosphate (DMAPP) into MBO, a volatile hemiterpene alcohol, in Pinus sabiniana. The result of enhanced phytol production in N. punctiforme, instead of MBO, could be explained by one of the 2 models: either the presence of a native prenyltransferase enzyme with a broad substrate specificity, or appropriation of a MBO synthase metabolic intermediate by a native geranyl diphosphate (GDP) synthase. In this work, an expression vector with an indigenous petE promoter for gene expression in the cyanobacterium N. punctiforme was constructed and MBO synthase gene expression was successfully shown using reverse transcriptase (RT)-PCR and SDS-PAGE. Gas chromatography--mass spectrophotometry (GC-MS) was performed to confirm phytol production from the transgenic N. punctiforme strains. We conclude that the expression of MBO synthase in N. punctiforme leads to overproduction of an economically important compound, phytol. This study provides insights about metabolic channeling of isoprenoids in cyanobacteria and also illustrates the challenges of bioengineering non-native hosts to produce economically important compounds.

  18. A Nostoc punctiforme Sugar Transporter Necessary to Establish a Cyanobacterium-Plant Symbiosis1[C][W

    Science.gov (United States)

    Ekman, Martin; Picossi, Silvia; Campbell, Elsie L.; Meeks, John C.; Flores, Enrique

    2013-01-01

    In cyanobacteria-plant symbioses, the symbiotic nitrogen-fixing cyanobacterium has low photosynthetic activity and is supplemented by sugars provided by the plant partner. Which sugars and cyanobacterial sugar uptake mechanism(s) are involved in the symbiosis, however, is unknown. Mutants of the symbiotically competent, facultatively heterotrophic cyanobacterium Nostoc punctiforme were constructed bearing a neomycin resistance gene cassette replacing genes in a putative sugar transport gene cluster. Results of transport activity assays using 14C-labeled fructose and glucose and tests of heterotrophic growth with these sugars enabled the identification of an ATP-binding cassette-type transporter for fructose (Frt), a major facilitator permease for glucose (GlcP), and a porin needed for the optimal uptake of both fructose and glucose. Analysis of green fluorescent protein fluorescence in strains of N. punctiforme bearing frt::gfp fusions showed high expression in vegetative cells and akinetes, variable expression in hormogonia, and no expression in heterocysts. The symbiotic efficiency of N. punctiforme sugar transport mutants was investigated by testing their ability to infect a nonvascular plant partner, the hornwort Anthoceros punctatus. Strains that were specifically unable to transport glucose did not infect the plant. These results imply a role for GlcP in establishing symbiosis under the conditions used in this work. PMID:23463784

  19. The non-metabolizable sucrose analog sucralose is a potent inhibitor of hormogonium differentiation in the filamentous cyanobacterium Nostoc punctiforme.

    Science.gov (United States)

    Splitt, Samantha D; Risser, Douglas D

    2016-03-01

    Nostoc punctiforme is a filamentous cyanobacterium which forms nitrogen-fixing symbioses with several different plants and fungi. Establishment of these symbioses requires the formation of motile hormogonium filaments. Once infected, the plant partner is thought to supply a hormogonium-repressing factor (HRF) to maintain the cyanobacteria in a vegetative, nitrogen-fixing state. Evidence implies that sucrose may serve as a HRF. Here, we tested the effects of sucralose, a non-metabolizable sucrose analog, on hormogonium differentiation. Sucralose inhibited hormogonium differentiation at a concentration approximately one-tenth that of sucrose. This result implies that: (1) sucrose, not a sucrose catabolite, is perceived by the cell and (2) inhibition is not due to a more general osmolarity-dependent effect. Additionally, both sucrose and sucralose induced the accrual of a polysaccharide sheath which bound specifically to the lectin ConA, indicating the presence of α-D-mannose and/or α-D-glucose. A ConA-specific polysaccharide was also found to be expressed in N. punctiforme colonies from tissue sections of the symbiotically grown hornwort Anthoceros punctatus. These findings imply that plant-derived sucrose or sucrose analogs may have multiple effects on N. punctiforme, including both repression of hormogonia and the induction of a polysaccharide sheath that may be essential to establish and maintain the symbiotic state.

  20. Characterization of two cation diffusion facilitators NpunF0707 and NpunF1794 in Nostoc punctiforme.

    Science.gov (United States)

    Hudek, L; Pearson, L; Michalczyk, A A; Bräu, L; Neilan, B A; Ackland, M L

    2015-11-01

    To characterize genes involved in maintaining homeostatic levels of zinc in the cyanobacterium Nostoc punctiforme. Metal efflux transporters play a central role in maintaining homeostatic levels of trace elements such as zinc. Sequence analyses of the N. punctiforme genome identified two potential cation diffusion facilitator (CDF) metal efflux transporters, Npun_F0707 (Cdf31) and Npun_F1794 (Cdf33). Deletion of either Cdf31or Cdf33 resulted in increased zinc retention over 3 h. Interestingly, Cdf31(-) and Cdf33(-) mutants showed no change in sensitivity to zinc exposure in comparison with the wild type, suggesting some compensatory capacity for the loss of each other. Using qRT-PCR, a possible interaction was observed between the two cdf's, where the Cdf31(-) mutant had a more profound effect on cdf33 expression than Cdf33(-) did on cdf31. Over-expression of Cdf31 and Cdf33 in ZntA(-) - and ZitB(-) -deficient Escherichia coli revealed function similarities between the ZntA and ZitB of E. coli and the cyanobacterial transporters. The data presented shed light on the function of two important transporters that regulate zinc homeostasis in N. punctiforme. This study shows for the first time the functional characterization of two cyanobacterial zinc efflux proteins belonging to the CDF family. © 2015 The Society for Applied Microbiology.

  1. Effect of Light Intensity and Photoperiod on Growth and Biochemical Composition of a Local Isolate of Nostoc calcicola.

    Science.gov (United States)

    Khajepour, Fateme; Hosseini, Seyed Abbas; Ghorbani Nasrabadi, Rasoul; Markou, Giorgos

    2015-08-01

    A study was conducted to investigate the effect of light intensity (21, 42, and 63 μmol photons m(-2) s(-1)) and photoperiod (8:16, 12:12, and 16:8 h light/dark) on the biomass production and its biochemical composition (total carotenoids, chlorophyll a, phycoerythrin (PE), phycocyanin (PC) and allophycocyanin (APC), total protein, and carbohydrates) of a local isolate of Nostoc calcicola. The results revealed that N. calcicola prefers dim light; however, the most of the levels of light intensity and photoperiod investigated did not have a significant impact on biomass production. Increasing light intensity biomass content of chlorophyll a, PE, PC, APC, and total protein decreased, while total carotenoids and carbohydrate increased. The same behavior was observed also when light duration (photoperiod) increased. The interaction effect of increasing light intensity and photoperiod resulted in an increase of carbohydrate and total carotenoids, and to the decrease of chlorophyll a, PE, PC, APC, and total protein content. The results indicate that varying the light regime, it is capable to manipulate the biochemical composition of the local isolate of N. calcicola, producing either valuable phycobiliproteins or proteins under low light intensity and shorter photoperiods, or producing carbohydrates and carotenoids under higher light intensities and longer photoperiods.

  2. Extracellular Polysaccharide Production in a Scytonemin-Deficient Mutant of Nostoc punctiforme Under UVA and Oxidative Stress.

    Science.gov (United States)

    Soule, Tanya; Shipe, Dexter; Lothamer, Justin

    2016-10-01

    Some cyanobacteria can protect themselves from ultraviolet radiation by producing sunscreen pigments. In particular, the sheath pigment scytonemin protects cells against long-wavelength UVA radiation and is only found in cyanobacteria which are capable of extracellular polysaccharide (EPS) production. The presence of a putative glycosyltransferase encoded within the scytonemin gene cluster, along with the localization of scytonemin and EPS to the extracellular sheath, prompted us to investigate the relationship between scytonemin and EPS production under UVA stress. In this study, it was hypothesized that there would be a relationship between the biosynthesis of scytonemin and EPS under both UVA and oxidative stress, since the latter is a by-product of UVA radiation. EPS production was measured following exposure of wild-type Nostoc punctiforme and the non-scytonemin-producing strain SCY59 to UVA and oxidative stress. Under UVA, SCY59 produced significantly more EPS than the unstressed controls and the wild type, while both strains produced more EPS under oxidative stress compared to the controls. The results suggest that EPS secretion occurs in response to the oxidative stress by-product of UVA rather than as a direct response to UVA radiation.

  3. Evolution of the tRNALeu (UAA Intron and Congruence of Genetic Markers in Lichen-Symbiotic Nostoc.

    Directory of Open Access Journals (Sweden)

    Ulla Kaasalainen

    Full Text Available The group I intron interrupting the tRNALeu UAA gene (trnL is present in most cyanobacterial genomes as well as in the plastids of many eukaryotic algae and all green plants. In lichen symbiotic Nostoc, the P6b stem-loop of trnL intron always involves one of two different repeat motifs, either Class I or Class II, both with unresolved evolutionary histories. Here we attempt to resolve the complex evolution of the two different trnL P6b region types. Our analysis indicates that the Class II repeat motif most likely appeared first and that independent and unidirectional shifts to the Class I motif have since taken place repeatedly. In addition, we compare our results with those obtained with other genetic markers and find strong evidence of recombination in the 16S rRNA gene, a marker widely used in phylogenetic studies on Bacteria. The congruence of the different genetic markers is successfully evaluated with the recently published software Saguaro, which has not previously been utilized in comparable studies.

  4. Hydrogen production and metal-dye bioremoval by a Nostoc linckia strain isolated from textile mill oxidation pond.

    Science.gov (United States)

    Mona, Sharma; Kaushik, Anubha; Kaushik, C P

    2011-02-01

    Biohydrogen production by Nostoc linckia HA-46, isolated from a textile-industry oxidation-pond was studied by varying light/dark period, pH, temperature and ratio of carbon-dioxide and argon in the gas-mixture. Hydrogen production rates were maximum under 18 h of light and 6 h of darkness, pH 8.0, 31°C, a CO(2):Ar ratio 2:10. Hydrogen production of the strain acclimatized to 20 mg/L of chromium/cobalt and 100 mg/L of Reactive red 198/crystal violet dye studied in N-supplemented/deficient medium was 6-10% higher in the presence of 1.5 g/L of NaNO(3). Rates of hydrogen production in the presence of dyes/metals by the strain (93-105 μmol/h/mg Chlorophyll) were significantly higher than in medium without metals/dyes serving as control (91.3 μmol/h/mg Chlorophyll). About 58-60% of the two metals and 35-73% of dyes were removed by cyanobacterium. Optimal conditions of temperature, pH and metals/dyes concentration for achieving high hydrogen production and wastewater treatment were found practically applicable as similar conditions are found in the effluent of regional textile-mills. Copyright © 2010 Elsevier Ltd. All rights reserved.

  5. Isolation and Purification of C-phycocyanin from Nostoc muscorum (Cyanophyceae and Cyanobacteria Exhibits Antimalarial Activity In vitro

    Directory of Open Access Journals (Sweden)

    Sukla Biswas

    2010-10-01

    Full Text Available The Phycobilin pigments are intensively fluorescent and water soluble. They are categorized into three types, such as pigments containing high, intermediate and low energies are Phycoerythrins (Phycoerythrocyanins, Phycocyanins and Allophycocyanins, respectively. Besides light harvesting, the Phycobiliproteins have shown industrial and biomedical importance. Among them, C-phycocyanin (C-PC has been considered to be the most preferred one. The present study was undertaken to evaluate the antimalarial activity of C-PC isolated from a Nitrogen-fixing Cyanobacterium and Nostoc muscorum. C- PC was extracted and purified by acetone extraction and ammonium sulfate precipitation and dialysis followed by amicon filtration. It was isolated as a~124 kDa water soluble protein molecule. It showed antimalarial activity in vitro against Chloroquine sensitive and resistant Plasmodium falciparum strains. Inhibitory concentrations at 50%, 90% and 95% were determined as 10.27±2.79, 53.53±6.26 and 73.78±6.92 µg/ml against the Chloroquine-sensitive strains; 10.37±1.43, 56.99±11.07 and 72.79±8.59 µg/ml against Chloroquine resistant of Plasmodium falciparum strains. C-PC was found to have antimalarial activity even at a concentration of 3.0 µg/ml. The possible mechanism might be relied on the destruction of polymerization of Haemozoin by binding of C-PC with Ferriprotoporphyrin-IX at the water surface of the plasma membrane.

  6. Differential physiological and biochemical responses of two cyanobacteria Nostoc muscorum and Phormidium foveolarum against oxyfluorfen and UV-B radiation.

    Science.gov (United States)

    Sheeba; Pratap Singh, Vijay; Kumar Srivastava, Prabhat; Mohan Prasad, Sheo

    2011-10-01

    In the present study, degree of tolerance and tolerance strategies of two paddy field cyanobacteria viz. Nostoc muscorum and Phormidium foveolarum against oxyfluorfen (10 and 20 μg ml(-1)) and UV-B (7.2 kJ m(-2)d(-1)) stress were investigated. Oxyfluorfen and UV-B decreased growth, photosynthesis, nutrient uptake, nitrate reductase, acid and alkaline phosphatase activities, which accompanied with the increase in the level of oxidative stress. However, growth was more affected in N. muscorum than P. foveolarum. Antioxidants exhibited differential responses against oxyfluorfen and UV-B stress. Ascorbate and proline levels were higher in P. foveolarum. A protein of 66 kDa was expressed in N. muscorum, however, it was absent in P. foveolarum than those of N. muscorum. Besides this, a protein of 29 kDa appeared in P. foveolarum under all the treatments, but it was present only in control cells of N. muscorum cells. Overall results indicated resistant nature of P. foveolarum against oxyfluorfen and UV-B stress in comparison to N. muscorum. Copyright © 2011 Elsevier Inc. All rights reserved.

  7. Cadmium toxicity to Microcystis aeruginosa PCC 7806 and its microcystin-lacking mutant.

    Directory of Open Access Journals (Sweden)

    Bin Huang

    Full Text Available The adverse effects of microcystin (MC produced by cyanobacteria have drawn considerable attention from the public. Yet it remains unclear whether MC confers any benefits to the cyanobacteria themselves. One suggested function of MC is complexation, which may influence the bioaccumulation and toxicity of trace metals. To test this hypothesis, we examined Cd toxicity to wild-type Microcystis aeruginosa PCC 7806 (WT and its MC-lacking mutant (MT under nutrient-enriched (+NP, phosphorus-limited (-P, and nitrogen-limited (-N conditions. The accumulation of Cd and the biochemical parameters associated with its detoxification [total phosphorus (TP, inorganic polyphosphate (Poly-P, and glutathione (GSH in the cells as well as intra- and extra-cellular carbohydrates] were quantified. Although the -P cyanobacteria accumulated less Cd than their +NP and -N counterparts, the different nutrient-conditioned cyanobacteria were similarly inhibited by similar free ion concentration of Cd in the medium ([Cd2+]F. Such good toxicity predictability of [Cd2+]F was ascribed to the synchronous decrease in the intracellular concentrations of Cd and TP. Nevertheless, Cd toxicity was still determined by the intracellular Cd to phosphorus ratio (Cd/P, in accordance with what has been reported in the literature. On the other hand, the concentrations of TP, Poly-P, and carbohydrates went up, but GSH concentration dropped down with the enhancement of [Cd2+]F, indicating their association with Cd detoxification. Although the inactivation of MC peptide synthetase gene had some nutrient and Cd concentration dependent effects on the parameters above, both cyanobacterial strains showed the same Cd accumulation ability and displayed similar Cd sensitivity. These results suggest that MC cannot affect metal toxicity either by regulating metal accumulation or by altering the detoxification ability of the cyanobacteria. Other possible functions of MC need to be further investigated.

  8. Antifungal compounds from cyanobacteria.

    Science.gov (United States)

    Shishido, Tânia K; Humisto, Anu; Jokela, Jouni; Liu, Liwei; Wahlsten, Matti; Tamrakar, Anisha; Fewer, David P; Permi, Perttu; Andreote, Ana P D; Fiore, Marli F; Sivonen, Kaarina

    2015-04-13

    Cyanobacteria are photosynthetic prokaryotes found in a range of environments. They are infamous for the production of toxins, as well as bioactive compounds, which exhibit anticancer, antimicrobial and protease inhibition activities. Cyanobacteria produce a broad range of antifungals belonging to structural classes, such as peptides, polyketides and alkaloids. Here, we tested cyanobacteria from a wide variety of environments for antifungal activity. The potent antifungal macrolide scytophycin was detected in Anabaena sp. HAN21/1, Anabaena cf. cylindrica PH133, Nostoc sp. HAN11/1 and Scytonema sp. HAN3/2. To our knowledge, this is the first description of Anabaena strains that produce scytophycins. We detected antifungal glycolipopeptide hassallidin production in Anabaena spp. BIR JV1 and HAN7/1 and in Nostoc spp. 6sf Calc and CENA 219. These strains were isolated from brackish and freshwater samples collected in Brazil, the Czech Republic and Finland. In addition, three cyanobacterial strains, Fischerella sp. CENA 298, Scytonema hofmanni PCC 7110 and Nostoc sp. N107.3, produced unidentified antifungal compounds that warrant further characterization. Interestingly, all of the strains shown to produce antifungal compounds in this study belong to Nostocales or Stigonematales cyanobacterial orders.

  9. Antifungal Compounds from Cyanobacteria

    Directory of Open Access Journals (Sweden)

    Tânia K. Shishido

    2015-04-01

    Full Text Available Cyanobacteria are photosynthetic prokaryotes found in a range of environments. They are infamous for the production of toxins, as well as bioactive compounds, which exhibit anticancer, antimicrobial and protease inhibition activities. Cyanobacteria produce a broad range of antifungals belonging to structural classes, such as peptides, polyketides and alkaloids. Here, we tested cyanobacteria from a wide variety of environments for antifungal activity. The potent antifungal macrolide scytophycin was detected in Anabaena sp. HAN21/1, Anabaena cf. cylindrica PH133, Nostoc sp. HAN11/1 and Scytonema sp. HAN3/2. To our knowledge, this is the first description of Anabaena strains that produce scytophycins. We detected antifungal glycolipopeptide hassallidin production in Anabaena spp. BIR JV1 and HAN7/1 and in Nostoc spp. 6sf Calc and CENA 219. These strains were isolated from brackish and freshwater samples collected in Brazil, the Czech Republic and Finland. In addition, three cyanobacterial strains, Fischerella sp. CENA 298, Scytonema hofmanni PCC 7110 and Nostoc sp. N107.3, produced unidentified antifungal compounds that warrant further characterization. Interestingly, all of the strains shown to produce antifungal compounds in this study belong to Nostocales or Stigonematales cyanobacterial orders.

  10. The response regulator Npun_F1278 is essential for scytonemin biosynthesis in the cyanobacterium Nostoc punctiforme ATCC 29133.

    Science.gov (United States)

    Naurin, Sejuti; Bennett, Janine; Videau, Patrick; Philmus, Benjamin; Soule, Tanya

    2016-08-01

    Following exposure to long-wavelength ultraviolet radiation (UVA), some cyanobacteria produce the indole-alkaloid sunscreen scytonemin. The genomic region associated with scytonemin biosynthesis in the cyanobacterium Nostoc punctiforme includes 18 cotranscribed genes. A two-component regulatory system (Npun_F1277/Npun_F1278) directly upstream from the biosynthetic genes was identified through comparative genomics and is likely involved in scytonemin regulation. In this study, the response regulator (RR), Npun_F1278, was evaluated for its ability to regulate scytonemin biosynthesis using a mutant strain of N. punctiforme deficient in this gene, hereafter strain Δ1278. Following UVA radiation, the typical stimulus to initiate scytonemin biosynthesis, Δ1278 was incapable of producing scytonemin. A phenotypic characterization of Δ1278 suggests that aside from the ability to produce scytonemin, the deletion of the Npun_F1278 gene does not affect the cellular morphology, cellular differentiation capability, or lipid-soluble pigment complement of Δ1278 compared to the wildtype. The mutant, however, had a slower specific growth rate under white light and produced ~2.5-fold more phycocyanin per cell under UVA than the wildtype. Since Δ1278 does not produce scytonemin, this study demonstrates that the RR gene, Npun_F1278, is essential for scytonemin biosynthesis in N. punctiforme. While most of the evaluated effects of this gene appear to be specific for scytonemin, this regulator may also influence the overall health of the cell and phycobiliprotein synthesis, directly or indirectly. This is the first study to identify a regulatory gene involved in the biosynthesis of the sunscreen scytonemin and posits a link between cell growth, pigment synthesis, and sunscreen production. © 2016 Phycological Society of America.

  11. Effects of UV-B Radiation and Periodic Desiccation on the Morphogenesis of the Edible Terrestrial Cyanobacterium Nostoc flagelliforme

    Science.gov (United States)

    Feng, Yan-Na; Zhang, Zhong-Chun; Feng, Jun-Li

    2012-01-01

    The terrestrial cyanobacterium Nostoc flagelliforme Berk. et M. A. Curtis has been a popular food and herbal ingredient for hundreds of years. To meet great market demand and protect the local ecosystem, for decades researchers have tried to cultivate N. flagelliforme but have failed to get macroscopic filamentous thalli. In this study, single trichomes with 50 to 200 vegetative cells were induced from free-living cells by low light and used to investigate the morphogenesis of N. flagelliforme under low UV-B radiation and periodic desiccation. Low-fluence-rate UV-B (0.1 W m−2) did not inhibit trichome growth; however, it significantly increased the synthesis of extracellular polysaccharides and mycosporine-like amino acids and promoted sheath formation outside the trichomes. Under low UV-B radiation, single trichomes developed into filamentous thalli more than 1 cm long after 28 days of cultivation, most of which grew separately in liquid BG11 medium. With periodic desiccation treatment, the single trichomes formed flat or banded thalli that grew up to 2 cm long after 3 months on solid BG11 medium. When trichomes were cultivated on solid BG11 medium with alternate treatments of low UV-B and periodic desiccation, dark and scraggly filamentous thalli that grew up to about 3 cm in length after 40 days were obtained. In addition, the cultivation of trichomes on nitrogen-deficient solid BG11 medium (BG110) suggested that nitrogen availability could affect the color and lubricity of newly developed thalli. This study provides promising techniques for artificial cultivation of N. flagelliforme in the future. PMID:22865081

  12. The response of antioxidant systems in Nostoc sphaeroides against UV-B radiation and the protective effects of exogenous antioxidants

    Science.gov (United States)

    Wang, Gaohong; Hu, Chunxiang; Li, Dunhai; Zhang, Delu; Li, Xiaoyan; Chen, Kun; Liu, Yongding

    UV radiation is one of many harmful factors found in space that are detrimental to organisms on earth in space exploration. In the present work, we examined the role of antioxidant system in Nostoc sphaeroides Kütz (Cyanobacterium) and the effects of exogenously applied antioxidant molecules on its photosynthetic rate under UV-B radiation. It was found that UV-B radiation promoted the activity of antioxidant system to protect photosystem II (PSII) and exogenously applied antioxidant: sodium nitroprusside (SNP) and N-acetylcysteine (NAC) had an obvious protection on PSII activity under UV-B radiation. The activity of superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6), peroxidase (POD, EC 1.11.1.7) and content of MDA (malondialdehyde) and ASC (ascorbate) were improved by 0.5 mM and 1 mM SNP, but 0.1 mM SNP decreased the activity of antioxidant system. Addition of exogenous NAC decreased the activity of SOD, POD, CAT and the content MDA and ASC. In contrast, exogenously applied NAC increased GSH content. The results suggest that exogenous SNP and NAC may protect algae by different mechanisms: SNP may play double roles as both sources of reactive free radicals as well as ROS scavengers in mediating the protective role of PSII on algae under UV-B radiation. On the other hand, NAC functions as an antioxidant or precursor of glutathione, which could protect PSII directly from UV-B radiation.

  13. SP-100 Program overview

    International Nuclear Information System (INIS)

    Truscello, V.C.

    1984-01-01

    The SP-100 Program is expected to go through three phases: technology assessment and advancement, ground testing, and flight qualification. Currently the program is in the two- to three-year technology assessment and advancement stage. Goals are to identify the space nuclear power system concept that best meets anticipated requirements of future space missions, assess the technical feasibility of that concept, and establish a cost and schedule for developing the concept. The SP-100 Project Office has begun the implementation activities needed to meet these goals. With regard to refractory alloys, a better data base will be required before we move ahead in the program from technology assessment to ground demonstration

  14. Persistence of Only a Minute Viable Population in Chlorotic Microcystis aeruginosa PCC 7806 Cultures Obtained by Nutrient Limitation.

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    Diogo de Abreu Meireles

    Full Text Available Cultures from the cyanobacterial strain Microcystis aeruginosa PCC 7806 submitted to nutrient limitation become chlorotic. When returned to nutrient rich conditions these cultures regain their green colour. The aim of this study was to verify whether the cells in these cultures could be considered resting stages allowing the survival of periods of nutrient starvation as has been reported for Synechococcus PCC 7942. The experiments with Microcystis were carried out in parallel with Synechococcus cultures to rule out the possibility that any results obtained with Microcystis were due to our particular experimental conditions. The results of the experiments with Synechococcus PCC 7942 cultures were comparable to the reported in the literature. For Microcystis PCC 7806 a different response was observed. Analysis of chlorotic Microcystis cultures by flow cytometry showed that the phenotype of the cells in the population was not homogenous: the amount of nucleic acids was about the same in all cells but only around one percent of the population emitted red autofluorescence indicating the presence of chlorophyll. Monitoring of the reversion of chlorosis by flow cytometry showed that the re-greening was most likely the result of the division of the small population of red autofluorescent cells originally present in the chlorotic cultures. This assumption was confirmed by analysing the integrity of the DNA and the membrane permeability of the cells of chlorotic cultures. Most of the DNA of these cultures was degraded and only the autofluorescent population of the chlorotic cultures showed membrane integrity. Thus, contrary to what has been reported for other cyanobacterial genera, most of the cells in chlorotic Microcystis cultures are not resting stages but dead. It is interesting to note that the red autofluorescent cells of green and chlorotic cultures obtained in double strength ASM-1 medium differ with respect to metabolism: levels of emission of

  15. Persistence of Only a Minute Viable Population in Chlorotic Microcystis aeruginosa PCC 7806 Cultures Obtained by Nutrient Limitation.

    Science.gov (United States)

    Meireles, Diogo de Abreu; Schripsema, Jan; Arnholdt, Andrea Cristina Vetö; Dagnino, Denise

    2015-01-01

    Cultures from the cyanobacterial strain Microcystis aeruginosa PCC 7806 submitted to nutrient limitation become chlorotic. When returned to nutrient rich conditions these cultures regain their green colour. The aim of this study was to verify whether the cells in these cultures could be considered resting stages allowing the survival of periods of nutrient starvation as has been reported for Synechococcus PCC 7942. The experiments with Microcystis were carried out in parallel with Synechococcus cultures to rule out the possibility that any results obtained with Microcystis were due to our particular experimental conditions. The results of the experiments with Synechococcus PCC 7942 cultures were comparable to the reported in the literature. For Microcystis PCC 7806 a different response was observed. Analysis of chlorotic Microcystis cultures by flow cytometry showed that the phenotype of the cells in the population was not homogenous: the amount of nucleic acids was about the same in all cells but only around one percent of the population emitted red autofluorescence indicating the presence of chlorophyll. Monitoring of the reversion of chlorosis by flow cytometry showed that the re-greening was most likely the result of the division of the small population of red autofluorescent cells originally present in the chlorotic cultures. This assumption was confirmed by analysing the integrity of the DNA and the membrane permeability of the cells of chlorotic cultures. Most of the DNA of these cultures was degraded and only the autofluorescent population of the chlorotic cultures showed membrane integrity. Thus, contrary to what has been reported for other cyanobacterial genera, most of the cells in chlorotic Microcystis cultures are not resting stages but dead. It is interesting to note that the red autofluorescent cells of green and chlorotic cultures obtained in double strength ASM-1 medium differ with respect to metabolism: levels of emission of red autofluorescence

  16. Comparison of bacterial community structures of terrestrial cyanobacterium Nostoc flagelliforme in three different regions of China using PCR-DGGE analysis.

    Science.gov (United States)

    Han, Pei-pei; Shen, Shi-gang; Jia, Shi-ru; Wang, Hui-yan; Zhong, Cheng; Tan, Zhi-lei; Lv, He-xin

    2015-07-01

    Filamentous Nostoc flagelliforme form colloidal complex, with beaded cells interacting with other bacteria embedded in the complex multilayer sheath. However, the species of bacteria in the sheath and the interaction between N. flagelliforme and associated bacteria remain unclear. In this study, PCR-denaturing gradient gel electrophoresis (DGGE) was used to investigate the bacterial communities of N. flagelliforme from three regions of China. DGGE patterns showed variations in all samples, exhibiting 25 discrete bands with various intensities. The diversity index analysis of bands profiles suggested the high similarity of bacterial communities to each other but also the dependence of microbial composition on each location. Phylogenetic affiliation indicated that the majority of the sequences obtained were affiliated with Actinobacteria, Cyanobacteria, Proteobacteria, Acidobacteria, Bacteroidetes, of which Cyanobacteria was dominant, followed the Proteobacteria. Members of the genus Nostoc were the most abundant in all samples. Rhizobiales and Actinobacteria were identified, whereas, Craurococcus, Caulobacter, Pseudomonas, Terriglobus and Mucilaginibacter were also identified at low levels. Through comparing the bacterial composition of N. flagelliforme from different regions, it was revealed that N. flagelliforme could facilitate the growth of other microorganisms including both autotrophic bacteria and heterotrophic ones and positively contributed to their harsh ecosystems. The results indicated N. flagelliforme played an important role in diversifying the microbial community composition and had potential application in soil desertification.

  17. Cryptosporidium sp. in lizards

    Czech Academy of Sciences Publication Activity Database

    Koudela, Břetislav; Modrý, D.

    1998-01-01

    Roč. 45, č. 1 (1998), s. 8 ISSN 1066-5234. [Cryptosporidium sp. in lazards. 01.01.1998-02.01.1998, Praha] R&D Projects: GA ČR GA508/95/0273; GA AV ČR IPP2020702 Subject RIV: fp - Other Medical Disciplines

  18. Komposit Nano TiO2 Dengan PCC, Zeolit atau Karbon Aktif Untuk Menurunkan Total Krom dan Zat Organik Pada Air Limbah Industri Penyamakan Kulit

    Directory of Open Access Journals (Sweden)

    Bumiarto Nugroho Jati

    2012-04-01

    Full Text Available Telah dilakukan penelitian untuk menurunkan total krom dan zat organik pada limbah industri penyamakan kulit dengan menggunakan nano TiO2 yang dikompositkan dengan adsorben karbon aktif, zeolit, dan precipitated calcium carbonate (PCC dalam suatu reaktor fotokatalitik yang disusun secara batch dan dilengkapi dengan 6 buah lampu UV dan magnetic stirrer. Penurunan kadar krom total diukur dengan menggunakan Atomic Absorption Spectro-photometer (AAS dan penurunan zat organik dianalisa dengan menggunakan titrasi permanganatometri. Hasil penelitian menunjukkan pengolahan terbaik untuk penurunan kadar krom total adalah dengan menggunakan komposit TiO2:PCC = 8:2 yang dapat menurunkan total krom hampir 100% pada menit ke-170 dengan konsentrasi awal 214,35 mg/L. Untuk penurunan kadar zat organik, pengolahan terbaik dengan menggunakan komposit TiO2:PCC = 9:1 yang dapat menurunkan kadar zat organik hingga 100% pada menit ke-180. 

  19. Photojournalism: the assaults of PCC in the pages of Folha and the Estadão Fotojornalismo: os ataques do PCC nas páginas da Folha e do Estadão

    Directory of Open Access Journals (Sweden)

    Paulo Cesar Boni

    2007-01-01

    Full Text Available This article addresses the photojournalistic coverage carried out by Folha de S.Paulo and O Estado de S. Paulo during the assaults of the First Command in the Capital (PCC in May 2006. The methods used here were those of technical deconstruction - to analyze the elements of photographic language in the construction of the message – and comparative analysis - to check the creation of meaning in those messages. Through these methodological procedures, it is inferred that Folha adopted a more sensationalistic feature with spectacularization of those images than Estadão, which adopted a more neutral and realistic posture in the view of the facts. Esse artigo aborda a cobertura fotojornalística realizada pela Folha de S.Paulo e pelo O Estado de S. Paulo durante os ataques do Primeiro Comando da Capital (PCC em maio de 2006. Os métodos utilizados foram o da desconstrução técnica – para análise dos elementos da linguagem fotográfica na construção da mensagem – e análise comparativa – para aferir a geração de sentido nas mensagens. Por esses procedimentos metodológicos, conclui que Folha assumiu um caráter mais sensacionalista, com espetacularização das imagens que o Estadão, que adotou uma postura mais neutra e realista diante dos fatos.

  20. Dissecting the hydrogenase expression and activity of transformed escherichia coli with the bidirectional NiFe-hydrogenase from synechocystis sp. PCC 6803

    International Nuclear Information System (INIS)

    Moon, Yu Ran; Lee, Min Hee; An, Byung Chull; Chung, Byung Yeoup; Kim, Jae Sung; Kim, Jin Hong; Park, Youn Il; Kim, Cha Soon

    2009-01-01

    Synechocystis bidirectional hydrogenase genes (hoxEFUYH) and their putative promoter regions and transformed into E. coli. The hox genes were transcribed in the E. coli cells carrying the vector construct of pCCIFOS::phox::hox or pCCIFOS::pT7::hox and translated into HoxEFUYH proteins, suggesting that the putative hox promoter can be constitutively activated in E. coli. Accordingly, the total hydrogenase activity was markedly increased up to 192% or 169% in the transformed cells, while the hydrogen uptake was decreased up to about 30% of the negative control. Although the gene expression of LexA, the only one transcription regulator proven for the hox genes, was substantially decreased in the pCCIFOS::phox::hox cells after γ-irradiation of 30 Gy, the expression levels of HoxEFUYH proteins were not altered significantly. Thus, it is also suggested that other transcription regulators as well as LexA might contribute to activation of the hox promoter in E. coli

  1. Association of Psb28 and Psb27 Proteins with PSII-PSI Supercomplexes upon Exposure of Synechocystis sp PCC 6803 to High Light

    Czech Academy of Sciences Publication Activity Database

    Bečková, Martina; Gardian, Zdenko; Yu, J.F.; Koník, Peter; Nixon, P. J.; Komenda, Josef

    2017-01-01

    Roč. 10, č. 1 (2017), s. 62-72 ISSN 1674-2052 R&D Projects: GA ČR GBP501/12/G055; GA MŠk(CZ) LO1416; GA MŠk(CZ) ED2.1.00/19.0392 Institutional support: RVO:61388971 ; RVO:60077344 Keywords : Psb28 protein s * photosystem I and II * Synechocystis Subject RIV: EE - Microbiology, Virology; CE - Biochemistry (BC-A) OBOR OECD: Microbiology; Biochemistry and molecular biology (BC-A) Impact factor: 8.827, year: 2016

  2. The ORF slr0091 of Synechocystis sp. PCC6803 encodes a high-light induced aldehyde dehydrogenase converting apocarotenals and alkanals

    KAUST Repository

    Trautmann, Danika; Beyer, Peter D.; Al-Babili, Salim

    2013-01-01

    Alh1 converts a wide range of apocarotenals and alkanals, with a preference for apocarotenals with defined chain lengths. As suggested by in vitro incubations and using engineered retinal-forming E. coli cells, we found that retinal is not a substrate

  3. Investigating the early stages of Photosystem II assembly in Synechocystis sp. PCC 6803: isolation of CP47 and CP43 complexe

    Czech Academy of Sciences Publication Activity Database

    Boehm, M.; Romero, E.; Reisinger, V.; Yu, J.; Komenda, Josef; Eichacker, L. A.; Dekker, J. P.; Nixon, P. J.

    2011-01-01

    Roč. 286, č. 17 (2011), 14812-14819 ISSN 0021-9258 R&D Projects: GA AV ČR IAA400200801 Institutional research plan: CEZ:AV0Z50200510 Keywords : ENERGY CHLOROPHYLL STATES * ANTENNA PROTEIN COMPLEX * OXYGEN-EVOLVING CENTER Subject RIV: EE - Microbiology, Virology Impact factor: 4.773, year: 2011

  4. Inactivation of the conserved open reading frame ycf34 of Synechocystis sp PCC 6803 interferes with the photosynthetic electron transport chain

    Czech Academy of Sciences Publication Activity Database

    Wallner, T.; Hagiwara, Y.; Bernát, G.; Sobotka, Roman; Reijerse, E. J.; Frankenberg-Dinkel, N.; Wilde, A.

    2012-01-01

    Roč. 1817, č. 11 (2012), s. 2016-2026 ISSN 0005-2728 R&D Projects: GA MŠk(CZ) ED2.1.00/03.0110 Institutional support: RVO:61388971 Keywords : Hypothetical chloroplast open reading frame * Iron-sulphur protein * Phycobilisome Subject RIV: EE - Microbiology, Virology Impact factor: 4.624, year: 2012

  5. A quantitative evaluation of ethylene production in the recombinant cyanobacterium Synechocystis sp PCC 6803 harboring the ethylene-forming enzyme by membrane inlet mass spectrometry

    Czech Academy of Sciences Publication Activity Database

    Zavřel, Tomáš; Knoop, H.; Steuer, R.; Jones, P. R.; Červený, Jan; Trtílek, M.

    2016-01-01

    Roč. 202, feb (2016), s. 142-151 ISSN 0960-8524 R&D Projects: GA MŠk(CZ) LO1415; GA ČR(CZ) GA15-17367S Institutional support: RVO:67179843 Keywords : biofuels * cyanobacteria * photobioreactor * MIMS * biotechnology Subject RIV: EH - Ecology, Behaviour Impact factor: 5.651, year: 2016

  6. The ORF slr0091 of Synechocystis sp. PCC6803 encodes a high-light induced aldehyde dehydrogenase converting apocarotenals and alkanals

    KAUST Repository

    Trautmann, Danika

    2013-07-05

    Oxidative cleavage of carotenoids and peroxidation of lipids lead to apocarotenals and aliphatic aldehydes called alkanals, which react with vitally important compounds, promoting cytotoxicity. Although many enzymes have been reported to deactivate alkanals by converting them into fatty acids, little is known about the mechanisms used to detoxify apocarotenals or the enzymes acting on them. Cyanobacteria and other photosynthetic organisms must cope with both classes of aldehydes. Here we report that the Synechocystis enzyme SynAlh1, encoded by the ORF slr0091, is an aldehyde dehydrogenase that mediates oxidation of both apocarotenals and alkanals into the corresponding acids. Using a crude lysate of SynAlh1-expressing Escherichia coli cells, we show that SynAlh1 converts a wide range of apocarotenals and alkanals, with a preference for apocarotenals with defined chain lengths. As suggested by in vitro incubations and using engineered retinal-forming E. coli cells, we found that retinal is not a substrate for SynAlh1, making involvement in Synechocystis retinoid metabolism unlikely. The transcript level of SynAlh1 is induced by high light and cold treatment, indicating a role in the stress response, and the corresponding gene is a constituent of a stress-related operon. The assumptions regarding the function of SynAlh are further supported by the surprisingly high homology to human and plant aldehyde dehydrogenase that have been assigned to aldehyde detoxification. SynAlh1 is the first aldehyde dehydrogenase that has been shown to form both apocarotenoic and fatty acids. This dual function suggests that its eukaryotic homologs may also be involved in apocarotenal metabolism, a function that has not been considered so far. Aldehyde dehydrogenases play an important role in detoxification of reactive aldehydes. Here, we report on a cyanbacterial enzyme capable in converting two classes of lipid-derived aldehydes, apocaotenals and alkanals. The corresponding gene is a constituent of a stress-related operon, and homology to eukaryotic enzymes points to a yet not considered possibility of their being involved in scavenging of apocarotenals. © 2013 FEBS.

  7. Small CAB-like proteins prevent formation of singlet oxygen in the damaged photosystem II complex of the cyanobacterium Synechocystis sp PCC 6803

    Czech Academy of Sciences Publication Activity Database

    Sinha, R. K.; Komenda, Josef; Knoppová, Jana; Sedlářová, M.; Pospíšil, P.

    2012-01-01

    Roč. 35, č. 4 (2012), s. 806-818 ISSN 0140-7791 R&D Projects: GA MŠk(CZ) ED2.1.00/03.0110; GA ČR(CZ) GAP501/11/0377 Institutional research plan: CEZ:AV0Z50200510 Keywords : oxidative stress * photoinhibition * reactive oxygen species Subject RIV: EE - Microbiology, Virology Impact factor: 5.135, year: 2012

  8. Final Techno-Economic Analysis of 550 MWe Supercritical PC Power Plant CO2 Capture with Linde-BASF Advanced PCC Technology

    Energy Technology Data Exchange (ETDEWEB)

    Bostick, Devin [Linde LLC, Murray Hill, NJ (United States); Stoffregen, Torsten [Linde AG Linde Engineering Division, Dresden (Germany); Rigby, Sean [BASF Corporation, Houston, TX (United States)

    2017-01-09

    This topical report presents the techno-economic evaluation of a 550 MWe supercritical pulverized coal (PC) power plant utilizing Illinois No. 6 coal as fuel, integrated with 1) a previously presented (for a subcritical PC plant) Linde-BASF post-combustion CO2 capture (PCC) plant incorporating BASF’s OASE® blue aqueous amine-based solvent (LB1) [Ref. 6] and 2) a new Linde-BASF PCC plant incorporating the same BASF OASE® blue solvent that features an advanced stripper interstage heater design (SIH) to optimize heat recovery in the PCC process. The process simulation and modeling for this report is performed using Aspen Plus V8.8. Technical information from the PCC plant is determined using BASF’s proprietary thermodynamic and process simulation models. The simulations developed and resulting cost estimates are first validated by reproducing the results of DOE/NETL Case 12 representing a 550 MWe supercritical PC-fired power plant with PCC incorporating a monoethanolamine (MEA) solvent as used in the DOE/NETL Case 12 reference [Ref. 2]. The results of the techno-economic assessment are shown comparing two specific options utilizing the BASF OASE® blue solvent technology (LB1 and SIH) to the DOE/NETL Case 12 reference. The results are shown comparing the energy demand for PCC, the incremental fuel requirement, and the net higher heating value (HHV) efficiency of the PC power plant integrated with the PCC plant. A comparison of the capital costs for each PCC plant configuration corresponding to a net 550 MWe power generation is also presented. Lastly, a cost of electricity (COE) and cost of CO2 captured assessment is shown illustrating the substantial cost reductions achieved with the Linde-BASF PCC plant utilizing the advanced SIH configuration in combination with BASF’s OASE® blue solvent technology as compared to the DOE/NETL Case 12 reference. The key factors contributing to the reduction of COE and the cost of CO2 captured

  9. A newly designed 45 to 60 mer oligonucleotide Agilent platform microarray for global gene expression studies of Synechocystis PCC6803: example salt stress experiment

    NARCIS (Netherlands)

    Aguirre von Wobeser, E.; Huisman, J.; Ibelings, B.; Matthijs, H.C.P.; Matthijs, H.C.P.

    2005-01-01

    A newly designed 45 to 60 mer oligonucleotide Agilent platform microarray for global gene expression studies of Synechocystis PCC6803: example salt stress experiment Eneas Aguirre-von-Wobeser 1, Jef Huisman1, Bas Ibelings2 and Hans C.P. Matthijs1 1 Universiteit van Amsterdam, Amsterdam, The

  10. Control of Branchionus sp. and Amoeba sp. in cultures of Arthrospira sp. Control de Branchionus sp. y Amoeba sp. en cultivos de Arthrospira sp.

    Directory of Open Access Journals (Sweden)

    Carlos Méndez

    2012-09-01

    Full Text Available Cultivation of cyanobacterium Arthrospira sp. has been developed in many countries for the production of proteins, pigments and other compounds. Outdoor mass cultures are often affected by biological contamination, drastically reducing productivity as far as bringing death. This study evaluates the control of Branchionus sp. and Amoeba sp. with two chemical compounds: urea (U and ammonium bicarbonate (AB, in laboratory conditions and outdoor mass culture of Arthrospira sp. The lethal concentration 100 (LC100 at 24 h for Branchionus sp. and Amoeba sp. determined was of 60-80 mg L-1 (U and 100-150 mg L-1 (AB. The average effective inhibition concentration for 50% of the population (IC50 in Arthrospira sp., after 72 h, was 80 mg L-1 (U and 150 mg L-1 (AB. The application of doses of 60 mg L-1 (U or 100 mg L-1 (AB in the outdoor mass culture of this contaminated microalga, completely inhibited grazing and did not affect the growth of Arthrospira sp. but rather promoted rapid recovery of algal density at levels prior to infestation. These compounds provided an economical and effective control of predators in cultures of Arthrospira sp.El cultivo de la cianobacteria Arthrospira sp. ha sido desarrollado en muchos países para la obtención de proteínas, pigmentos y otros compuestos. Cultivo que a nivel industrial se ve afectado frecuentemente por contaminación biológica, reduciendo drásticamente la productividad hasta causar la muerte. Este estudio evalúa el control de Branchionus sp. y de Amoeba sp. con dos compuestos químicos, la urea (U y bicarbonato de amonio (AB en cultivos de Arthrospira sp. La concentración letal 100 (LC100 determinada a las 24 h para Branchionus sp. y Amoeba sp. fue de 60-80 mg L-1 (U y 100-150 mg L-1 (AB. La concentración media de inhibición efectiva, después de 72 h, para el 50% de la población (IC50 en Arthrospira fue de 80 mg L-1 (U y 150 mg L-1 (AB. La aplicación de dosis de 60 mg L-1 (U ó 100 mg L-1 (AB en

  11. Development of a low-cost oxy-hydrogen bio-fuel cell for generation of electricity using Nostoc as a source of hydrogen

    Energy Technology Data Exchange (ETDEWEB)

    Sangeeta Dawar; Behera, B.K. [Maharshi Dayanand University, Rohtak (India). Dept. of Biosciences; Prasanna Mohanty [Jawaharlal Nehru University, New Delhi (India). School of Life Sciences

    1998-10-10

    An oxy-hydrogen bio-fuel cell, based on a carbon-carbon electrode has been fabricated. The electrode pellets were prepared by taking carbon powder mixed with polyvinylalcohol as a binder. The anode was charged with Co-Al spinel mixed oxide at 700{sup o}C, 30% KOH acted as an electrolyte. For the cyanobacterial bioreactor, a potential heterocystous blue green alga of Nostoc spp. has been used for hydrogen production and electrical energy generation. Various nutrient enrichment techniques are employed to increase the hydrogen generation efficiency of the algae. One litre free cell algal reactor attached to the fuel cell, at the anode end for hydrogen gas input, generated about 300 mV of voltage and 100 mA of current. Our present findings on the development of a low cost fuel cell with high efficiency of current output may be helpful in commercializing this technology. (author)

  12. Isolation Of PS II Nanoparticles And Oxygen Evolution Studies In Synechococcus Spp. PCC 7942 Under Heavy Metal Stress

    Science.gov (United States)

    Ahmad, Iffat Zareen; Sundaram, Shanthy; Tripathi, Ashutosh; Soumya, K. K.

    2009-06-01

    The effect of heavy metals was seen on the oxygen evolution pattern of a unicellular, non-heterocystous cyanobacterial strain of Synechococcus spp. PCC 7942. It was grown in a BG-11 medium supplemented with heavy metals, namely, nickel, copper, cadmium and mercury. Final concentrations of the heavy metal solution used in the culture were 0.1, 0.4 and 1 μM. All the experiments were performed in the exponential phase of the culture. Oxygen-evolving photosystem II (PS II) particles were purified from Synechococcus spp. PCC 7942 by a single-step Ni2+-affinity column chromatography after solubilization of thylakoid membranes with sucrose monolaurate. Oxygen evolution was measured with Clark type oxygen electrode fitted with a circulating water jacket. The light on the surface of the vessel was 10 w/m2. The cultures were incubated in light for 15 minutes prior to the measurement of oxygen evolution. Oxygen evolution was measured in assay mixture containing phosphate buffer (pH-7.5, 0.1 M) in the presence of potassium ferricyanide as the electron acceptor. The preparation from the control showed a high oxygen-evolving activity of 2, 300-2, 500 pmol O2 (mg Chl)-1 h-1 while the activity was decreased in the cultures grown with heavy metals. The inhibition of oxygen evolution shown by the organism in the presence of different metals was in the order Hg>Ni>Cd>Cu. Such heavy metal resistant strains will find application in the construction of PS II- based biosensors for the monitoring of pollutants.

  13. PRR11 regulates late-S to G2/M phase progression and induces premature chromatin condensation (PCC)

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Chundong; Zhang, Ying; Li, Yi; Zhu, Huifang; Wang, Yitao; Cai, Wei [Department of Biochemistry and Molecular Biology, Chongqing Medical University, Chongqing 400016 (China); Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing 400016 (China); Zhu, Jiang [Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing 400016 (China); Ozaki, Toshinori [Laboratory of DNA Damage Signaling, Chiba Cancer Center Research Institute, 666-2 Nitona, Chuohku, Chiba 260-8717 (Japan); Bu, Youquan, E-mail: buyqcn@aliyun.com [Department of Biochemistry and Molecular Biology, Chongqing Medical University, Chongqing 400016 (China); Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing 400016 (China)

    2015-03-13

    Recently, we have demonstrated that proline-rich protein 11 (PRR11) is a novel tumor-related gene product likely implicated in the regulation of cell cycle progression as well as lung cancer development. However, its precise role in cell cycle progression remains unclear. In the present study, we have further investigated the expression pattern and functional implication of PRR11 during cell cycle in detail in human lung carcinoma-derived H1299 cells. According to our immunofluorescence study, PRR11 was expressed largely in cytoplasm, the amount of PRR11 started to increase in the late S phase, and was retained until just before mitotic telophase. Consistent with those observations, siRNA-mediated knockdown of PRR11 caused a significant cell cycle arrest in the late S phase. Intriguingly, the treatment with dNTPs further augmented PRR11 silencing-mediated S phase arrest. Moreover, knockdown of PRR11 also resulted in a remarkable retardation of G2/M progression, and PRR11-knockdown cells subsequently underwent G2 phase cell cycle arrest accompanied by obvious mitotic defects such as multipolar spindles and multiple nuclei. In addition, forced expression of PRR11 promoted the premature Chromatin condensation (PCC), and then proliferation of PRR11-expressing cells was massively attenuated and induced apoptosis. Taken together, our current observations strongly suggest that PRR11, which is strictly regulated during cell cycle progression, plays a pivotal role in the regulation of accurate cell cycle progression through the late S phase to mitosis. - Highlights: • PRR11 started to increase in the late S phase and was retained until just before mitotic telophase. • PRR11-knockdown caused a significant cell cycle arrest in the late S phase and G2 phase. • The treatment with dNTPs further augmented PRR11 silencing-mediated S phase arrest. • PRR11-knockdown led to multipolar spindles and multiple nuclei. • Forced expression of PRR11 promoted the PCC and inhibited

  14. PRR11 regulates late-S to G2/M phase progression and induces premature chromatin condensation (PCC)

    International Nuclear Information System (INIS)

    Zhang, Chundong; Zhang, Ying; Li, Yi; Zhu, Huifang; Wang, Yitao; Cai, Wei; Zhu, Jiang; Ozaki, Toshinori; Bu, Youquan

    2015-01-01

    Recently, we have demonstrated that proline-rich protein 11 (PRR11) is a novel tumor-related gene product likely implicated in the regulation of cell cycle progression as well as lung cancer development. However, its precise role in cell cycle progression remains unclear. In the present study, we have further investigated the expression pattern and functional implication of PRR11 during cell cycle in detail in human lung carcinoma-derived H1299 cells. According to our immunofluorescence study, PRR11 was expressed largely in cytoplasm, the amount of PRR11 started to increase in the late S phase, and was retained until just before mitotic telophase. Consistent with those observations, siRNA-mediated knockdown of PRR11 caused a significant cell cycle arrest in the late S phase. Intriguingly, the treatment with dNTPs further augmented PRR11 silencing-mediated S phase arrest. Moreover, knockdown of PRR11 also resulted in a remarkable retardation of G2/M progression, and PRR11-knockdown cells subsequently underwent G2 phase cell cycle arrest accompanied by obvious mitotic defects such as multipolar spindles and multiple nuclei. In addition, forced expression of PRR11 promoted the premature Chromatin condensation (PCC), and then proliferation of PRR11-expressing cells was massively attenuated and induced apoptosis. Taken together, our current observations strongly suggest that PRR11, which is strictly regulated during cell cycle progression, plays a pivotal role in the regulation of accurate cell cycle progression through the late S phase to mitosis. - Highlights: • PRR11 started to increase in the late S phase and was retained until just before mitotic telophase. • PRR11-knockdown caused a significant cell cycle arrest in the late S phase and G2 phase. • The treatment with dNTPs further augmented PRR11 silencing-mediated S phase arrest. • PRR11-knockdown led to multipolar spindles and multiple nuclei. • Forced expression of PRR11 promoted the PCC and inhibited

  15. SP mountain data analysis

    Science.gov (United States)

    Rawson, R. F.; Hamilton, R. E.; Liskow, C. L.; Dias, A. R.; Jackson, P. L.

    1981-01-01

    An analysis of synthetic aperture radar data of SP Mountain was undertaken to demonstrate the use of digital image processing techniques to aid in geologic interpretation of SAR data. These data were collected with the ERIM X- and L-band airborne SAR using like- and cross-polarizations. The resulting signal films were used to produce computer compatible tapes, from which four-channel imagery was generated. Slant range-to-ground range and range-azimuth-scale corrections were made in order to facilitate image registration; intensity corrections were also made. Manual interpretation of the imagery showed that L-band represented the geology of the area better than X-band. Several differences between the various images were also noted. Further digital analysis of the corrected data was done for enhancement purposes. This analysis included application of an MSS differencing routine and development of a routine for removal of relief displacement. It was found that accurate registration of the SAR channels is critical to the effectiveness of the differencing routine. Use of the relief displacement algorithm on the SP Mountain data demonstrated the feasibility of the technique.

  16. Systems Level Approaches to Understanding and Manipulating Heterocyst Differentiation in Nostoc Punctiforme: Sites of Hydrogenase and Nitrogenase Synthesis and Activity

    Energy Technology Data Exchange (ETDEWEB)

    Meeks, John C. [University of California, Davis, CA (United States)

    2015-04-02

    Heterocysts are specialized cells that establish a physiologically low oxygen concentration; they function as the sites of oxygen-sensitive nitrogen fixation and hydrogen metabolism in certain filamentous cyanobacteria. They are present at a frequency of less than 10% of the cells and singly in a nonrandom spacing pattern in the filaments. The extent of differential gene expression during heterocyst differentiation was defined by DNA microarray analysis in wild type and mutant cultures of Nostoc punctiforme. The results in wild-type cultures identified two groups of genes; approximately 440 that are unique to heterocyst formation and function, and 500 that respond positively and negatively to the transient stress of nitrogen starvation. Nitrogen fixation is initiated within 24 h after induction, but the cultures require another 24 h before growth is reinitiated. Microarray analyses were conducted on strains with altered expression of three genes that regulate the presence and spacing of heterocysts in the filaments; loss of function or over expression of these genes increases the heterocyst frequency 2 to 3 fold compared to the wild-type. Mutations in the genes hetR and hetF result in the inability to differentiate heterocysts, whereas over expression of each gene individually yields multiple contiguous heterocysts at sites in the filaments; they are positive regulatory elements. Mutation of the gene patN results in an increase in heterocysts frequency, but, in this case, the heterocysts are singly spaced in the filaments with a decrease in the number of vegetative cells in the interval between heterocysts; this is a negative regulatory element. However, over expression of patN resulted in the wild-type heterocyst frequency and spacing pattern. Microarray results indicated HetR and HetF influence the transcription of a common set of about 395 genes, as well as about 350 genes unique to each protein. HetR is known to be a transcriptional regulator and HetF is

  17. Metabolic Engineering of Light and Dark Biochemical Pathways in Wild-Type and Mutant Strains of Synechocystis PCC 6803 for Maximal, 24-Hour Production of Hydrogen Gas

    Energy Technology Data Exchange (ETDEWEB)

    Ely, Roger L.; Chaplen, Frank W.R.

    2014-03-11

    This project used the cyanobacterial species Synechocystis PCC 6803 to pursue two lines of inquiry, with each line addressing one of the two main factors affecting hydrogen (H2) production in Synechocystis PCC 6803: NADPH availability and O2 sensitivity. H2 production in Synechocystis PCC 6803 requires a very high NADPH:NADP+ ratio, that is, the NADP pool must be highly reduced, which can be problematic because several metabolic pathways potentially can act to raise or lower NADPH levels. Also, though the [NiFe]-hydrogenase in PCC 6803 is constitutively expressed, it is reversibly inactivated at very low O2 concentrations. Largely because of this O2 sensitivity and the requirement for high NADPH levels, a major portion of overall H2 production occurs under anoxic conditions in the dark, supported by breakdown of glycogen or other organic substrates accumulated during photosynthesis. Also, other factors, such as N or S limitation, pH changes, presence of other substances, or deletion of particular respiratory components, can affect light or dark H2 production. Therefore, in the first line of inquiry, under a number of culture conditions with wild type (WT) Synechocystis PCC 6803 cells and a mutant with impaired type I NADPH-dehydrogenase (NDH-1) function, we used H2 production profiling and metabolic flux analysis, with and without specific inhibitors, to examine systematically the pathways involved in light and dark H2 production. Results from this work provided rational bases for metabolic engineering to maximize photobiological H2 production on a 24-hour basis. In the second line of inquiry, we used site-directed mutagenesis to create mutants with hydrogenase enzymes exhibiting greater O2 tolerance. The research addressed the following four tasks: 1. Evaluate the effects of various culture conditions (N, S, or P limitation; light/dark; pH; exogenous organic carbon) on H2 production profiles of WT cells and an NDH-1 mutant; 2. Conduct metabolic flux analyses for

  18. SP-100 Test Site

    International Nuclear Information System (INIS)

    Cox, C.M.; Mahaffey, M.K.; Miller, W.C.

    1988-01-01

    Preparatory activities are well under way at Hanford to convert the 309 Containment Building and its associated service wing to a 2.5 MWt nuclear test facility for the SP-100 Ground Engineering System (GES) test. Preliminary design is complete, encompassing facility modifications, a secondary heat transport system, a large vacuum system to enclose the high temperature reactor, a test assembly cell and handling system, control and data processing systems, and safety and auxiliary systems. The design makes extensive use of existing equipment to minimize technical risk and cost. Refurbishment of this equipment is 75% complete. The facility has been cleared of obstructing equipment from its earlier reactor test. Current activities are focusing on definitive design and preparation of the Preliminary Safety Analysis Report (PSAR) aimed at procurement and construction approvals and schedules to achieve reactor criticality by January 1992. 6 refs

  19. A comparison of G2 phase radiation-induced chromatid break kinetics using calyculin-PCC with those obtained using colcemid block.

    Science.gov (United States)

    Bryant, Peter E; Mozdarani, Hossein

    2007-09-01

    To study the possible influence of cell-cycle delay on cells reaching mitosis during conventional radiation-induced chromatid break experiments using colcemid as a blocking agent, we have compared the chromatid break kinetics following a single dose of gamma rays (0.75 Gy) in metaphase CHO cells using calyculin-induced premature chromosome condensation (PCC), with those using colcemid block. Calyculin-induced PCC causes very rapid condensation of G2 cell chromosomes without the need for a cell to progress to mitosis, hence eliminating any effect of cell-cycle checkpoint on chromatid break frequency. We found that the kinetics of the exponential first-order decrease in chromatid breaks with time after irradiation was similar (not significantly different) between the two methods of chromosome condensation. However, use of the calyculin-PCC technique resulted in a slightly increased rate of disappearance of chromatid breaks and thus higher frequencies of breaks at 1.5 and 2.5 h following irradiation. We also report on the effect of the nucleoside analogue ara A on chromatid break kinetics using the two chromosome condensation techniques. Ara A treatment of cells abrogated the decrease in chromatid breaks with time, both using the calyculin-PCC and colcemid methods. We conclude that cell-cycle delay may be a factor determining the absolute frequency of chromatid breaks at various times following irradiation of cells in G2 phase but that the first-order disappearance of chromatid breaks with time and its abrogation by ara A are not significantly influenced by the G2 checkpoint.

  20. A Putative O-Linked β-N-Acetylglucosamine Transferase Is Essential for Hormogonium Development and Motility in the Filamentous Cyanobacterium Nostoc punctiforme.

    Science.gov (United States)

    Khayatan, Behzad; Bains, Divleen K; Cheng, Monica H; Cho, Ye Won; Huynh, Jessica; Kim, Rachelle; Omoruyi, Osagie H; Pantoja, Adriana P; Park, Jun Sang; Peng, Julia K; Splitt, Samantha D; Tian, Mason Y; Risser, Douglas D

    2017-05-01

    Most species of filamentous cyanobacteria are capable of gliding motility, likely via a conserved type IV pilus-like system that may also secrete a motility-associated polysaccharide. In a subset of these organisms, motility is achieved only after the transient differentiation of hormogonia, which are specialized filaments that enter a nongrowth state dedicated to motility. Despite the fundamental importance of hormogonia to the life cycles of many filamentous cyanobacteria, the molecular regulation of hormogonium development is largely undefined. To systematically identify genes essential for hormogonium development and motility in the model heterocyst-forming filamentous cyanobacterium Nostoc punctiforme , a forward genetic screen was employed. The first gene identified using this screen, designated ogtA , encodes a putative O-linked β- N -acetylglucosamine transferase (OGT). The deletion of ogtA abolished motility, while ectopic expression of ogtA induced hormogonium development even under hormogonium-repressing conditions. Transcription of ogtA is rapidly upregulated (1 h) following hormogonium induction, and an OgtA-GFPuv fusion protein localized to the cytoplasm. In developing hormogonia, accumulation of PilA but not HmpD is dependent on ogtA Reverse transcription-quantitative PCR (RT-qPCR) analysis indicated equivalent levels of pilA transcript in the wild-type and Δ ogtA mutant strains, while a reporter construct consisting of the intergenic region in the 5' direction of pilA fused to gfp produced lower levels of fluorescence in the Δ ogtA mutant strain than in the wild type. The production of hormogonium polysaccharide in the Δ ogtA mutant strain is reduced compared to that in the wild type but comparable to that in a pilA deletion strain. Collectively, these results imply that O -GlcNAc protein modification regulates the accumulation of PilA via a posttranscriptional mechanism in developing hormogonia. IMPORTANCE Filamentous cyanobacteria are among

  1. Bradysia sp. em morangueiro Bradysia sp. in strawberry

    Directory of Open Access Journals (Sweden)

    Bernadete Radin

    2009-04-01

    Full Text Available No trabalho, relatam-se os primeiros registros de Bradysia sp. (Insecta: Diptera: Sciaridae em morangueiro (Fragaria x ananassa Duch., cultivado no Município de Eldorado do Sul, RS. O cultivo foi realizado em sacolas com três metros de comprimento, preenchidas com substrato composto de casca de arroz e turfa, dispostas horizontalmente sobre bancadas de madeira, em ambiente protegido. A presença de Bradysia sp. foi observada na segunda quinzena de agosto de 2005. Neste trabalho, estão descritos os sintomas apresentados no morangueiro pela praga, prováveis conseqüências sobre o aparecimento de doenças e uma breve descrição morfológica da Bradysia sp., adulto e fase larval.This paper describes the first record of Bradysia sp. (Insecta; Diptera; Sciaridae in strawberry (Fragaria x ananassa, cultivated in the city of Eldorado do Sul, RS, Brazil. Strawberry was planted in plastic bags filled with a mixture of burnt rice hulls and peat and cultivated in a greenhouse. The presence of Bradysia sp was noticed in the second fortnight of August, 2005. The symptoms in strawberry and the probable consequences in terms of disease arising were described in the present study, as well as the morphological characterization of Bradysia sp. and its illustrations.

  2. Yersinia pekkanenii sp. nov.

    Science.gov (United States)

    Murros-Kontiainen, Anna; Johansson, Per; Niskanen, Taina; Fredriksson-Ahomaa, Maria; Korkeala, Hannu; Björkroth, Johanna

    2011-10-01

    The taxonomic position of three strains from water, soil and lettuce samples was studied by using a polyphasic taxonomic approach. The strains were reported to lack the virulence-encoding genes inv and virF in a previous study. Controversially, API 20 E and some other phenotypic tests suggested that the strains belong to Yersinia pseudotuberculosis, which prompted this polyphasic taxonomic study. In both the phylogenetic analyses of four housekeeping genes (glnA, gyrB, recA and HSP60) and numerical analyses of HindIII and EcoRI ribopatterns, the strains formed a separate group within the genus Yersinia. Analysis of the 16S rRNA gene sequences showed that the strains were related to Yersinia aldovae and Yersinia mollaretii, but DNA-DNA hybridization analysis differentiated them from these species. Based on the results of the phylogenetic and DNA-DNA hybridization analyses, a novel species, Yersinia pekkanenii sp. nov., is proposed. The type strain is ÅYV7.1KOH2(T) ( = DSM 22769(T)  = LMG 25369(T)).

  3. Antimicrobial peptides from Capsicum sp.

    African Journals Online (AJOL)

    ajl yemi

    2011-12-30

    Dec 30, 2011 ... Key words: Antimicrobial peptides, Capsicum sp, Capsicum chinense, chili pepper, agronomical options, ..... of this human activity is resumed by the simple phrase: produce .... It will be interesting to scale the AMPs extraction.

  4. Metabolic engineering of cyanobacteria for photosynthetic 3-hydroxypropionic acid production from CO2 using Synechococcus elongatus PCC 7942.

    Science.gov (United States)

    Lan, Ethan I; Chuang, Derrick S; Shen, Claire R; Lee, Annabel M; Ro, Soo Y; Liao, James C

    2015-09-01

    Photosynthetic conversion of CO2 to chemicals using cyanobacteria is an attractive approach for direct recycling of CO2 to useful products. 3-Hydroxypropionic acid (3 HP) is a valuable chemical for the synthesis of polymers and serves as a precursor to many other chemicals such as acrylic acid. 3 HP is naturally produced through glycerol metabolism. However, cyanobacteria do not possess pathways for synthesizing glycerol and converting glycerol to 3 HP. Furthermore, the latter pathway requires coenzyme B12, or an oxygen sensitive, coenzyme B12-independent enzyme. These characteristics present major challenges for production of 3 HP using cyanobacteria. To overcome such difficulties, we constructed two alternative pathways in Synechococcus elongatus PCC 7942: a malonyl-CoA dependent pathway and a β-alanine dependent pathway. Expression of the malonyl-CoA dependent pathway genes (malonyl-CoA reductase and malonate semialdehyde reductase) enabled S. elongatus to synthesize 3 HP to a final titer of 665 mg/L. β-Alanine dependent pathway expressing S. elongatus produced 3H P to final titer of 186 mg/L. These results demonstrated the feasibility of converting CO2 into 3 HP using cyanobacteria. Copyright © 2015 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  5. Carbon-free production of 2-deoxy-scyllo-inosose (DOI) in cyanobacterium Synechococcus elongatus PCC 7942.

    Science.gov (United States)

    Watanabe, Satoru; Ozawa, Hiroaki; Kato, Hiroaki; Nimura-Matsune, Kaori; Hirayama, Toshifumi; Kudo, Fumitaka; Eguchi, Tadashi; Kakinuma, Katsumi; Yoshikawa, Hirofumi

    2018-01-01

    Owing to their photosynthetic capabilities, there is increasing interest in utilizing cyanobacteria to convert solar energy into biomass. 2-Deoxy-scyllo-inosose (DOI) is a valuable starting material for the benzene-free synthesis of catechol and other benzenoids. DOI synthase (DOIS) is responsible for the formation of DOI from d-glucose-6-phosphate (G6P) in the biosynthesis of 2-deoxystreptamine-containing aminoglycoside antibiotics such as neomycin and butirosin. DOI fermentation using a recombinant Escherichia coli strain has been reported, although a carbon source is necessary for high-yield DOI production. We constructed DOI-producing cyanobacteria toward carbon-free and sustainable DOI production. A DOIS gene derived from the butirosin producer strain Bacillus circulans (btrC) was introduced and expressed in the cyanobacterium Synechococcus elongatus PCC 7942. We ultimately succeeded in producing 400 mg/L of DOI in S. elongatus without using a carbon source. DOI production by cyanobacteria represents a novel and efficient approach for producing benzenoids from G6P synthesized by photosynthesis.

  6. Alcohol-tolerant mutants of cyanobacterium Synechococcus elongatus PCC 7942 obtained by single-cell mutant screening system.

    Science.gov (United States)

    Arai, Sayuri; Hayashihara, Kayoko; Kanamoto, Yuki; Shimizu, Kazunori; Hirokawa, Yasutaka; Hanai, Taizo; Murakami, Akio; Honda, Hiroyuki

    2017-08-01

    Enhancement of alcohol tolerance in microorganisms is an important strategy for improving bioalcohol productivity. Although cyanobacteria can be used as a promising biocatalyst to produce various alcohols directly from CO 2 , low productivity, and low tolerance against alcohols are the main issues to be resolved. Nevertheless, to date, a mutant with increasing alcohol tolerance has rarely been reported. In this study, we attempted to select isopropanol (IPA)-tolerant mutants of Synechococcus elongatus PCC 7942 using UV-C-induced random mutagenesis, followed by enrichment of the tolerant candidates in medium containing 10 g/L IPA and screening of the cells with a high growth rate in the single cell culture system in liquid medium containing 10 g/L IPA. We successfully acquired the most tolerant strain, SY1043, which maintains the ability to grow in medium containing 30 g/L IPA. The photosynthetic oxygen-evolving activities of SY1043 were almost same in cells after 72 h incubation under light with or without 10 g/L IPA, while the activity of the wild-type was remarkably decreased after the incubation with IPA. SY1043 also showed higher tolerance to ethanol, 1-butanol, isobutanol, and 1-pentanol than the wild type. These results suggest that SY1043 would be a promising candidate to improve alcohol production using cyanobacteria. Biotechnol. Bioeng. 2017;114: 1771-1778. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  7. Growth of nutrient-replete Microcystis PCC 7806 cultures is inhibited by an extracellular signal produced by chlorotic cultures.

    Science.gov (United States)

    Dagnino, Denise; de Abreu Meireles, Diogo; de Aquino Almeida, João Carlos

    2006-01-01

    The frequency of cyanobacterial blooms has been increasing all over the world. These blooms are often toxic and have become a serious health problem. The aim of this work was to search for population density control mechanisms that could inhibit the proliferation of the toxic bloom-forming genus Microcystis. Microcystis PCC 7806 cultured for long periods in liquid ASM-1 medium loses its characteristic green colour. When a medium of chlorotic cultures is added to a nutrient-replete culture, cell density increase is drastically reduced when compared with controls. Inhibition of cell proliferation occurs in Microcystis cultures from any growth stage and was not strain-specific, but other genera tested showed no response. Investigations on the mechanism of growth inhibition showed that cultures treated with the conditioned medium acquired a pale colour, with pigment concentration similar to that found in chlorotic cultures. Ultrastructural examination showed that the conditioned medium induced thylakoid membrane disorganization, typical of chlorotic cells, in nutrient-replete cultures. An active extract was obtained and investigations showed that activity was retained after heating and after addition of an apolar solvent. This indicates that activity of the conditioned medium from chlorotic cells results from non-protein, apolar compound(s).

  8. Fluorescence Spectral Properties of All4261 Binding with Phycocyanobilin in E.Coli

    Science.gov (United States)

    Ma, Q.; Zheng, X. J.; Zhou, Z.; Zhou, N.; Zhao, K. H.; Zhou, M.

    2014-07-01

    Cyanobacteriochromes (CBCRs) are chromophorylated proteins that acting as sensory photoreceptors in cyanobacteria. Based on the bioinformatics of All4261 in Nostoc sp. PCC7120, All4261 is a CBCR apoprotein composed of GAF domains in the N-terminal region. Via polymerase chain reaction with specific primers, All4261 was amplified with genome DNA of Nostoc sp. PCC7120 as template and then subcloned into the expression vector pET30(a+). To survey the fluorescence spectral properties, All4261 was coexpressed with the plasmid that catalyzes phycocyanobilin (PCB) biosynthesis, pACYC-ho1-pcyA, in E.coli BL21. Fluorescence emission spectra and excitation spectra showed that chromophorylated cells containing All4261-PCB had a fluorescence emission peak at 645 nm and a fluorescence excitation peak at 550 nm, but no reversible photoconversion. In order to identify the binding site of PCB in All4261, we obtained three variants All4261(C296L), All4261(C328A), and All4261(C339L), via sitedirected mutagenesis. The binding site was identified as C339 based on the lack of PCB binding of All4261(C339L).

  9. A common transport system for methionine, L-methionine-DL-sulfoximine (MSX), and phosphinothricin (PPT) in the diazotrophic cyanobacterium Nostoc muscorum.

    Science.gov (United States)

    Singh, Arvind Kumar; Syiem, Mayashree B; Singh, Rajkumar S; Adhikari, Samrat; Rai, Amar Nath

    2008-05-01

    We present evidence, for the first time, of the occurrence of a transport system common for amino acid methionine, and methionine/glutamate analogues L-methionine-DL-sulfoximine (MSX) and phosphinothricin (PPT) in cyanobacterium Nostoc muscorum. Methionine, which is toxic to cyanobacterium, enhanced its nitrogenase activity at lower concentrations. The cyanobacterium showed a biphasic pattern of methionine uptake activity that was competitively inhibited by the amino acids alanine, isoleucine, leucine, phenylalanine, proline, valine, glutamine, and asparagine. The methionine/glutamate analogue-resistant N. muscorum strains (MSX-R and PPT-R strains) also showed methionine-resistant phenotype accompanied by a drastic decrease in 35S methionine uptake activity. Treatment of protein extracts from these mutant strains with MSX and PPT reduced biosynthetic glutamine synthetase (GS) activity only in vitro and not in vivo. This finding implicated that MSX- and PPT-R phenotypes may have arisen due to a defect in their MSX and PPT transport activity. The simultaneous decrease in methionine uptake activity and in vitro sensitivity toward MSX and PPT of GS protein in MSX- and PPT-R strains indicated that methionine, MSX, and PPT have a common transport system that is shared by other amino acids as well in N. muscorum. Such information can become useful for isolation of methionine-producing cyanobacterial strains.

  10. Complementary UV-Absorption of Mycosporine-like Amino Acids and Scytonemin is Responsible for the UV-Insensitivity of Photosynthesis in Nostoc flagelliforme

    Science.gov (United States)

    Ferroni, Lorenzo; Klisch, Manfred; Pancaldi, Simonetta; Häder, Donat-Peter

    2010-01-01

    Mycosporine-like amino acids (MAAs) and scytonemin are UV-screening compounds that have presumably appeared early in the history of life and are widespread in cyanobacteria. Natural colonies of the UV-insensitive Nostoc flagelliforme were found to be especially rich in MAAs (32.1 mg g DW−1), concentrated in the glycan sheath together with scytonemin. MAAs are present in the form of oligosaccharide-linked molecules. Photosystem II activity, measured using PAM fluorescence and oxygen evolution, was used as a most sensitive physiological parameter to analyse the effectiveness of UV-protection. Laboratory experiments were performed under controlled conditions with a simulated solar radiation specifically deprived of UV-wavebands with cut-off filters (295, 305, 320, 345 and 395 nm). The UV-insensitivity of N. flagelliforme was found to cover the whole UV-A (315–400 nm) and UV-B (280–320 nm) range and is almost certainly due to the complementary UV-absorption of MAAs and scytonemin. The experimental approach used is proposed to be suitable for the comparison of the UV-protection ability in organisms that differ in their complement of UV-sunscreen compounds. Furthermore, this study performed with a genuinely terrestrial organism points to the relevance of marine photoprotective compounds for life on Earth, especially for the colonization of terrestrial environments. PMID:20161974

  11. Complementary UV-Absorption of Mycosporine-like Amino Acids and Scytonemin is Responsible for the UV-Insensitivity of Photosynthesis in Nostoc flagelliforme

    Directory of Open Access Journals (Sweden)

    Donat-Peter Häder

    2010-01-01

    Full Text Available Mycosporine-like amino acids (MAAs and scytonemin are UV-screening compounds that have presumably appeared early in the history of life and are widespread in cyanobacteria. Natural colonies of the UV-insensitive Nostoc flagelliforme were found to be especially rich in MAAs (32.1 mg g DW-1, concentrated in the glycan sheath together with scytonemin. MAAs are present in the form of oligosaccharide-linked molecules. Photosystem II activity, measured using PAM fluorescence and oxygen evolution, was used as a most sensitive physiological parameter to analyse the effectiveness of UV-protection. Laboratory experiments were performed under controlled conditions with a simulated solar radiation specifically deprived of UV-wavebands with cut-off filters (295, 305, 320, 345 and 395 nm. The UV-insensitivity of N. flagelliforme was found to cover the whole UV-A (315–400 nm and UV-B (280–320 nm range and is almost certainly due to the complementary UV-absorption of MAAs and scytonemin. The experimental approach used is proposed to be suitable for the comparison of the UV-protection ability in organisms that differ in their complement of UV-sunscreen compounds. Furthermore, this study performed with a genuinely terrestrial organism points to the relevance of marine photoprotective compounds for life on Earth, especially for the colonization of terrestrial environments.

  12. Molecular Cloning and Biochemical Characterization of the Iron Superoxide Dismutase from the Cyanobacterium Nostoc punctiforme ATCC 29133 and Its Response to Methyl Viologen-Induced Oxidative Stress.

    Science.gov (United States)

    Moirangthem, Lakshmipyari Devi; Ibrahim, Kalibulla Syed; Vanlalsangi, Rebecca; Stensjö, Karin; Lindblad, Peter; Bhattacharya, Jyotirmoy

    2015-12-01

    Superoxide dismutase (SOD) detoxifies cell-toxic superoxide radicals and constitutes an important component of antioxidant machinery in aerobic organisms, including cyanobacteria. The iron-containing SOD (SodB) is one of the most abundant soluble proteins in the cytosol of the nitrogen-fixing cyanobacterium Nostoc punctiforme ATCC 29133, and therefore, we investigated its biochemical properties and response to oxidative stress. The putative SodB-encoding open reading frame Npun_R6491 was cloned and overexpressed in Escherichia coli as a C-terminally hexahistidine-tagged protein. The purified recombinant protein had a SodB specific activity of 2560 ± 48 U/mg protein at pH 7.8 and was highly thermostable. The presence of a characteristic iron absorption peak at 350 nm, and its sensitivity to H2O2 and azide, confirmed that the SodB is an iron-containing SOD. Transcript level of SodB in nitrogen-fixing cultures of N. punctiforme decreased considerably (threefold) after exposure to an oxidative stress-generating herbicide methyl viologen for 4 h. Furthermore, in-gel SOD activity analysis of such cultures grown at increasing concentrations of methyl viologen also showed a loss of SodB activity. These results suggest that SodB is not the primary scavenger of superoxide radicals induced by methyl viologen in N. punctiforme.

  13. Enhancement of stability of L-tryptophan dehydrogenase from Nostoc punctiforme ATCC29133 and its application to L-tryptophan assay.

    Science.gov (United States)

    Matsui, Daisuke; Okazaki, Seiji; Matsuda, Motoki; Asano, Yasuhisa

    2015-02-20

    Microbial NAD(+)-dependent L-tryptophan dehydrogenase (TrpDH, EC1.4.1.19), which catalyzes the reversible oxidative deamination and the reductive amination between L-tryptophan and indole-3-pyruvic acid, was found in the scytonemin biosynthetic pathway of Nostoc punctiforme ATCC29133. The TrpDH exhibited high specificity toward L-tryptophan, but its instability was a drawback for L-tryptophan determination. The mutant enzyme TrpDH L59F/D168G/A234D/I296N with thermal stability was obtained by screening of Escherichia coli transformants harboring various mutant genes, which were generated by error-prone PCR using complementation in an L-tryptophan auxotroph of E. coli. The specific activity and stability of this mutant enzyme were higher than those of the wild type enzyme. We also revealed here that in these four mutation points, the two amino acid residues Asp168 and Ile296 contributed to increase the enzyme stability, and the Leu59, Ala234 residues to increase its specific activity. Growth of the strain harboring the gene of above 4 point mutated enzyme was accelerated by the enhanced performance. In the present study, we demonstrated that TrpDH L59F/D168G/A234D/I296N was available for determination of L-tryptophan in human plasma. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Crystallization and preliminary X-ray crystallographic analysis of the GluR0 ligand-binding core from Nostoc punctiforme

    International Nuclear Information System (INIS)

    Lee, Jun Hyuck; Park, Soo Jeong; Rho, Seong-Hwan; Im, Young Jun; Kim, Mun-Kyoung; Kang, Gil Bu; Eom, Soo Hyun

    2005-01-01

    The GluR0 ligand-binding core from N. punctiforme was expressed, purified and crystallized in the presence of l-glutamate. A diffraction data set was collected to a resolution of 2.1 Å. GluR0 from Nostoc punctiforme (NpGluR0) is a bacterial homologue of the ionotropic glutamate receptor. The ligand-binding core of NpGluR0 was crystallized at 294 K using the hanging-drop vapour-diffusion method. The l-glutamate-complexed crystal belongs to space group C222 1 , with unit-cell parameters a = 78.0, b = 145.1, c = 132.1 Å. The crystals contain three subunits in the asymmetric unit, with a V M value of 2.49 Å 3 Da −1 . The diffraction limit of the l-glutamate complex data set was 2.1 Å using synchrotron X-ray radiation at beamline BL-4A of the Pohang Accelerator Laboratory (Pohang, Korea)

  15. The ZntA-like NpunR4017 plays a key role in maintaining homeostatic levels of zinc in Nostoc punctiforme.

    Science.gov (United States)

    Hudek, L; Bräu, L; Michalczyk, A A; Neilan, B A; Meeks, J C; Ackland, M L

    2015-12-01

    Analysis of cellular response to zinc exposure provides insights into how organisms maintain homeostatic levels of zinc that are essential, while avoiding potentially toxic cytosolic levels. Using the cyanobacterium Nostoc punctiforme as a model, qRT-PCR analyses established a profile of the changes in relative mRNA levels of the ZntA-like zinc efflux transporter NpunR4017 in response to extracellular zinc. In cells treated with 18 μM of zinc for 1 h, NpunR4017 mRNA levels increased by up to 1300 % above basal levels. The accumulation and retention of radiolabelled (65)Zn by NpunR4107-deficient and overexpressing strains were compared to wild-type levels. Disruption of NpunR4017 resulted in a significant increase in zinc accumulation up to 24 % greater than the wild type, while cells overexpressing NpunR4107 accumulated 22 % less than the wild type. Accumulation of (65)Zn in ZntA(-) Escherichia coli overexpressing NpunR4017 was reduced by up to 21 %, indicating the capacity for NpunR4017 to compensate for the loss of ZntA. These findings establish the newly identified NpunR4017 as a zinc efflux transporter and a key transporter for maintaining zinc homeostasis in N. punctiforme.

  16. A high constitutive catalase activity confers resistance to methyl viologen-promoted oxidative stress in a mutant of the cyanobacterium Nostoc punctiforme ATCC 29133.

    Science.gov (United States)

    Moirangthem, Lakshmipyari Devi; Bhattacharya, Sudeshna; Stensjö, Karin; Lindblad, Peter; Bhattacharya, Jyotirmoy

    2014-04-01

    A spontaneous methyl viologen (MV)-resistant mutant of the nitrogen-fixing cyanobacterium Nostoc punctiforme ATCC 29133 was isolated and the major enzymatic antioxidants involved in combating MV-induced oxidative stress were evaluated. The mutant displayed a high constitutive catalase activity as a consequence of which, the intracellular level of reactive oxygen species in the mutant was lower than the wild type (N. punctiforme) in the presence of MV. The superoxide dismutase (SOD) activity that consisted of a SodA (manganese-SOD) and a SodB (iron-SOD) was not suppressed in the mutant following MV treatment. The mutant was, however, characterised by a lower peroxidase activity compared with its wild type, and its improved tolerance to externally added H₂O₂ could only be attributed to enhanced catalase activity. Furthermore, MV-induced toxic effects on the wild type such as (1) loss of photosynthetic performance assessed as maximal quantum yield of photosystem II, (2) nitrogenase inactivation, and (3) filament fragmentation and cell lysis were not observed in the mutant. These findings highlight the importance of catalase in preventing MV-promoted oxidative damage and cell death in the cyanobacterium N. punctiforme. Such oxidative stress resistant mutants of cyanobacteria are likely to be a better source of biofertilisers, as they can grow and fix nitrogen in an unhindered manner in agricultural fields that are often contaminated with the herbicide MV, also commonly known as paraquat.

  17. Characterization and in vivo regulon determination of an ECF sigma factor and its cognate anti-sigma factor in Nostoc punctiforme.

    Science.gov (United States)

    Bell, Nicole; Lee, Jamie J; Summers, Michael L

    2017-04-01

    Based on primary sequence comparisons and genomic context, Npun_F4153 (SigG)/Npun_F4154 (SapG) of the cyanobacterium Nostoc punctiforme were hypothesized to encode an ECF sigma factor/anti-sigma factor pair. Transcription of sigG increased in heterocysts and akinetes, and after EDTA treatment. Interaction between SigG and the predicted cytoplasmic domain of SapG was observed in vitro. A SigG-GFP translational fusion protein localized to the periphery of vegetative cells in vivo, but lost this association following heat stress. A sigG mutant was unable to survive envelope damage caused by heat or EDTA, but was able to form functional heterocysts. Akinetes in the mutant strain appeared normal, but these cultures were less resistant to lysozyme and cold treatments than those of the wild-type strain. The SigG in vivo regulon was determined before and during akinete differentiation using DNA microarray analysis, and found to include multiple genes with putative association to the cell envelope. Mapped promoters common to both arrays enabled identification of a SigG promoter-binding motif that was supported in vivo by reporter studies, and in vitro by run-off transcription experiments. These findings support SigG/SapG as a sigma/anti-sigma pair involved in repair of envelope damage resulting from exogenous sources or cellular differentiation. © 2017 John Wiley & Sons Ltd.

  18. Expression and characterization of a recombinant psychrophilic γ-carbonic anhydrase (NcoCA) identified in the genome of the Antarctic cyanobacteria belonging to the genus Nostoc.

    Science.gov (United States)

    De Luca, Viviana; Del Prete, Sonia; Vullo, Daniela; Carginale, Vincenzo; Di Fonzo, Pietro; Osman, Sameh M; AlOthman, Zeid; Supuran, Claudiu T; Capasso, Clemente

    2016-10-01

    Carbonic anhydrases (CAs, EC 4.2.1.1) catalyze the CO2 hydration/dehydration reversible reaction: CO2 + H2O ⇄ [Formula: see text] + H(+). Living organisms encode for at least six distinct genetic families of such