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Sample records for cellulolytic actinomycete thermobifida

  1. Hydrophobic nature and effects of culture conditions on biofilm formation by the cellulolytic actinomycete Thermobifida fusca

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    Almaris N. Alonso

    2015-09-01

    Full Text Available Thermobifida fusca produces a firmly attached biofilm on nutritive and non-nutritive surfaces, such as cellulose, glass, plastic, metal and Teflon®. The ability to bind to surfaces has been suggested as a competitive advantage for microbes in soil environments. Results of previous investigations indicated that a Gram-positive cellulolytic soil bacteria, Cellulomonas uda, a facultative aerobe, specifically adhered to nutritive surfaces forming biofilms, but cells did not colonize non-nutritive surfaces. Cell surface hydrophobicity has been implicated in the interactions between bacteria and the adhesion to surfaces. It was recently described that the cellulolytic actinomycete T. fusca cells hydrophobicity was measured and compared to the cellulolytic soil bacteria C. uda. Also, T. fusca biofilm formation on non-nutritive surface, such as polyvinyl chloride, was examined by testing various culture ingredients to determine a possible trigger mechanism for biofilm formation. Experimental results showed that partitioning of bacterial cells to various hydrocarbons was higher in T. fusca cells than in C. uda. The results of this study suggest that the attachment to multiple surfaces by T. fusca could depend on nutrient availability, pH, salt concentrations, and the higher hydrophobic nature of bacterial cells. Possibly, these characteristics may confer T. fusca a selective advantage to compete and survive among the many environments it thrives.

  2. Genome Sequence and Analysis of the Soil Cellulolytic ActinomyceteThermobifida fusca

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    Lykidis, Athanasios; Mavromatis, Konstantinos; Ivanova, Natalia; Anderson, Iain; Land, Miriam; DiBartolo, Genevieve; Martinez, Michele; Lapidus, Alla; Lucas, Susan; Copeland, Alex; Richardson, Paul; Wilson,David B.; Kyrpides, Nikos

    2007-02-01

    Thermobifida fusca is a moderately thermophilic soilbacterium that belongs to Actinobacteria. 3 It is a major degrader ofplant cell walls and has been used as a model organism for the study of 4secreted, thermostable cellulases. The complete genome sequence showedthat T. fusca has a 5 single circular chromosome of 3642249 bp predictedto encode 3117 proteins and 65 RNA6 species with a coding densityof 85percent. Genome analysis revealed the existence of 29 putative 7glycoside hydrolases in addition to the previously identified cellulasesand xylanases. The 8 glycosyl hydrolases include enzymes predicted toexhibit mainly dextran/starch and xylan 9 degrading functions. T. fuscapossesses two protein secretion systems: the sec general secretion 10system and the twin-arginine translocation system. Several of thesecreted cellulases have 11 sequence signatures indicating theirsecretion may be mediated by the twin-arginine12 translocation system. T.fusca has extensive transport systems for import of carbohydrates 13coupled to transcriptional regulators controlling the expression of thetransporters and14 glycosylhydrolases. In addition to providing anoverview of the physiology of a soil 15 actinomycete, this study presentsinsights on the transcriptional regulation and secretion of16 cellulaseswhich may facilitate the industrial exploitation of thesesystems.

  3. Isolation of Cellulolytic Actinomycetes from Marine Sediments

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    Veiga, Manuel; Esparis, Azucena; Fabregas, Jaime

    1983-01-01

    The cellulolytic activity of 36 actinomycetes strains isolated from marine sediments was investigated by the cellulose-azure method. Approximately 50% of the isolates exhibited various degrees of cellulolytic activity.

  4. Actinomycetes mycetoma

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    Sumati Hogade

    2011-01-01

    Full Text Available Mycetoma is a chronic infection, frequently seen in tropical and sub-tropical countries and is considered as an occupational disease. Nocardia species though it can infect immunocompetent individuals, it most commonly affects immunocompromised patients. A 50-year-old male, farmer presented to our hospital with serosanguineous discharging swelling over the dorsum of right foot. We have isolated Nocardia asteroides from the tissue sample. Speciation of this isolate was carried out based on phenotypic methods. Hereby we report a case of Actinomycetes Mycetoma in an immunocompetent individual.

  5. Actinomycetes: A Source of Lignocellulolytic Enzymes

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    Saini, Anita; Aggarwal, Neeraj K.; Sharma, Anuja; Yadav, Anita

    2015-01-01

    Lignocellulose is the most abundant biomass on earth. Agricultural, forest, and agroindustrial activities generate tons of lignocellulosic wastes annually, which present readily procurable, economically affordable, and renewable feedstock for various lignocelluloses based applications. Lignocelluloses are the focus of present decade researchers globally, in an attempt to develop technologies based on natural biomass for reducing dependence on expensive and exhaustible substrates. Lignocellulolytic enzymes, that is, cellulases, hemicellulases, and lignolytic enzymes, play very important role in the processing of lignocelluloses which is prerequisite for their utilization in various processes. These enzymes are obtained from microorganisms distributed in both prokaryotic and eukaryotic domains including bacteria, fungi, and actinomycetes. Actinomycetes are an attractive microbial group for production of lignocellulose degrading enzymes. Various studies have evaluated the lignocellulose degrading ability of actinomycetes, which can be potentially implemented in the production of different value added products. This paper is an overview of the diversity of cellulolytic, hemicellulolytic, and lignolytic actinomycetes along with brief discussion of their hydrolytic enzyme systems involved in biomass modification. PMID:26793393

  6. Indigenous cellulolytic and hemicellulolytic bacteria enhanced rapid co-composting of lignocellulose oil palm empty fruit bunch with palm oil mill effluent anaerobic sludge.

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    Zainudin, Mohd Huzairi Mohd; Hassan, Mohd Ali; Tokura, Mitsunori; Shirai, Yoshihito

    2013-11-01

    The composting of lignocellulosic oil palm empty fruit bunch (OPEFB) with continuous addition of palm oil mill (POME) anaerobic sludge which contained nutrients and indigenous microbes was studied. In comparison to the conventional OPEFB composting which took 60-90 days, the rapid composting in this study can be completed in 40 days with final C/N ratio of 12.4 and nitrogen (2.5%), phosphorus (1.4%), and potassium (2.8%), respectively. Twenty-seven cellulolytic bacterial strains of which 23 strains were closely related to Bacillus subtilis, Bacillus firmus, Thermobifida fusca, Thermomonospora spp., Cellulomonas sp., Ureibacillus thermosphaericus, Paenibacillus barengoltzii, Paenibacillus campinasensis, Geobacillus thermodenitrificans, Pseudoxanthomonas byssovorax which were known as lignocellulose degrading bacteria and commonly involved in lignocellulose degradation. Four isolated strains related to Exiguobacterium acetylicum and Rhizobium sp., with cellulolytic and hemicellulolytic activities. The rapid composting period achieved in this study can thus be attributed to the naturally occurring cellulolytic and hemicellulolytic strains identified.

  7. Cloning, Expression and Biochemical Characterization of Endomannanases from Thermobifida Species Isolated from Different Niches.

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    Ákos Tóth

    Full Text Available Thermobifidas are thermotolerant, compost inhabiting actinomycetes which have complex polysaccharide hydrolyzing enzyme systems. The best characterized enzymes of these hydrolases are cellulases from T. fusca, while other important enzymes especially hemicellulases are not deeply explored. To fill this gap we cloned and investigated endomannanases from those reference strains of the Thermobifida genus, which have published data on other hydrolases (T. fusca TM51, T. alba CECT3323, T. cellulosilytica TB100T and T. halotolerans YIM90462T. Our phylogenetic analyses of 16S rDNA and endomannanase sequences revealed that T. alba CECT3323 is miss-classified; it belongs to the T. fusca species. The cloned and investigated endomannanases belong to the family of glycosyl hydrolases 5 (GH5, their size is around 50 kDa and they are modular enzymes. Their catalytic domains are extended by a C-terminal carbohydrate binding module (CBM of type 2 with a 23-25 residues long interdomain linker region consisting of Pro, Thr and Glu/Asp rich repetitive tetrapeptide motifs. Their polypeptide chains exhibit high homology, interdomain sequence, which don't show homology to each other, but all of them are built up from 3-6 times repeated tetrapeptide motifs (PTDP-Tc, TEEP-Tf, DPGT-Th. All of the heterologously expressed Man5A enzymes exhibited activity only on mannan. The pH optima of Man5A enzymes from T. halotolerans, T. cellulosilytica and T. fusca are slightly different (7.0, 7.5 and 8.0, respectively while their temperature optima span within the range of 70-75°C. The three endomannanases exhibited very similar kinetic performances on LBG-mannan substrate: 0.9-1.7mM of KM and 80-120 1/sec of turnover number. We detected great variability in heat stability at 70°C, which was influenced by the presence of Ca2+. The investigated endomannanases might be important subjects for studying the structure/function relation behind the heat stability and for industrial

  8. Soil actinomycetes in the National Forest Park in northeastern China

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    Shirokikh, I. G.; Shirokikh, A. A.

    2017-01-01

    The taxonomic and functional structure of actinomycete complexes in the litters and upper horizons of the soils under an artificial coniferous-broad-leaved forest located around the town of Chanchun (Tszilin province, PRC). The complex of actinomycetes included representatives of the Streptomyces, Micromonospora, Streptosporangium, and Streptoverticillium genera and oligosporous forms. In the actinomycete complexes, streptomycetes prevailed in the abundance (61-95%) and frequency of occurrence (100%). In the parcels of Korean pine ( Pinus koraiensis) and Mongolian oak ( Quercus mongolica), streptomycetes of 19 species from 8 series and 4 sections were isolated. The most representative, as in European forest biomes, was the Cinereus Achromogenes series. A distinguishing feature of the streptomycete complex in the biomes studied was the high participation of species from the Imperfectus series. The verification of the functional activity of natural isolates made it possible to reveal strains with high antagonistic and cellulolytic abilities. A high similarity of actinomycete complexes was found in Eurasian forest ecosystems remote from each other, probably due to the similarity of plant polymers decomposable by actinomycetes.

  9. Cellulolytic enzyme compositions and uses thereof

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    Iyer, Prashant; Gaspar, Armindo Ribiero; Croonenberghs, James; Binder, Thomas P.

    2017-07-25

    The present invention relates enzyme composition comprising a cellulolytic preparation and an acetylxylan esterase (AXE); and the used of cellulolytic enzyme compositions for hydrolyzing acetylated cellulosic material. Finally the invention also relates to processes of producing fermentation products from acetylated cellulosic materials using a cellulolytic enzyme composition of the invention.

  10. Polypeptide from a cellulolytic fungus having cellulolytic enhancing activity

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    Brown, Kimberly (Elk Grove, CA); Harris, Paul (Carnation, WA); Zaretsky, Elizabeth (Reno, NV); Re, Edward (Davis, CA); Vlasenko, Elena (Davis, CA); McFarland, Keith (Davis, CA); Lopez de Leon, Alfredo (Davis, CA)

    2008-04-22

    The present invention relates to isolated polypeptides having cellulolytic enhancing activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods for producing and using the polypeptides.

  11. Cellulolytic Microorganisms from Thermal Environments

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    Vishnivetskaya, Tatiana A [ORNL; Raman, Babu [ORNL; Phelps, Tommy Joe [ORNL; Podar, Mircea [ORNL; Elkins, James G [ORNL

    2012-01-01

    Thermal, anaerobic environments rich in decaying plant material are a potential source of novel cellulolytic bacteria. Samples collected from geothermal aquifers in the Yellowstone National Park (YNP) were used to select for cellulolytic thermophiles. Laboratory enrichments on dilute-acid pretreated plant biomass (switchgrass, Populus), and crystalline cellulose (Avicel) resulted in the isolation of 247 environmental clones. The majority of individual clones were affiliated with the cellulolytic bacteria of phylum Firmicutes, followed by xylanolytic and saccharolytic members of the phylum Dictyoglomi. Among the Firmicutes, the clones were affiliated with the genera Caldicellulosiruptor (54.4%), Caloramator (11.5%), Thermoanaerobacter (8.8%), Thermovenabulum (4.1%), and Clostridium (2.0%). From established anaerobic thermophilic enrichments a total of 81 single strains of the genera Caldicellulosiruptor (57%) and Thermoanaerobacter (43%) were isolated. With continuous flow enrichment on Avicel, increases in the relative abundance of Caloramator sp. was observed over clones detected from the Caldicellulosiruptor. Complex communities of interacting microorganisms bring about cellulose decomposition in nature, therefore using up-to-date approaches may yield novel cellulolytic microorganisms with high activity and a rapid rate of biomass conversion to biofuels.

  12. Short-chain aliphatic ester synthesis using Thermobifida fusca cutinase.

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    Su, Lingqia; Hong, Ruoyu; Guo, Xiaojie; Wu, Jing; Xia, Yongmei

    2016-09-01

    Short-chain aliphatic esters are commonly used as fruit flavorings in the food industry. In this study, Thermobifida fusca (T. fusca) cutinase was used for the synthesis of aliphatic esters, and the maximum yield of ethyl caproate reached 99.2% at a cutinase concentration of 50U/ml, 40°C, and water content of 0.5%, representing the highest ester yield to date. The cutinase-catalyzed esterification displayed strong tolerance for water content (up to 8%) and acid concentration (up to 0.8M). At substrate concentrations ⩽0.8M, the ester yield remained above 80%. Moreover, ester yields of more than 98% and 95% were achieved for acids of C3-C8 and alcohols of C1-C6, respectively, indicating extensive chain length selectivity of the cutinase. These results demonstrate the superior ability of T. fusca cutinase to catalyze the synthesis of short-chain esters. This study provides the basis for industrial production of short-chain esters using T. fusca cutinase.

  13. Not so monofunctional-a case of thermostable Thermobifida fusca catalase with peroxidase activity

    NARCIS (Netherlands)

    Lončar, Nikola; Fraaije, Marco W

    Thermobifida fusca is a mesothermophilic organism known for its ability to degrade plant biomass and other organics, and it was demonstrated that it represents a rich resource of genes encoding for potent enzymes for biocatalysis. The thermostable catalase from T. fusca has been cloned and

  14. Not so monofunctional-a case of thermostable Thermobifida fusca catalase with peroxidase activity

    NARCIS (Netherlands)

    Lončar, Nikola; Fraaije, Marco W

    2014-01-01

    Thermobifida fusca is a mesothermophilic organism known for its ability to degrade plant biomass and other organics, and it was demonstrated that it represents a rich resource of genes encoding for potent enzymes for biocatalysis. The thermostable catalase from T. fusca has been cloned and overexpre

  15. Not so monofunctional-a case of thermostable Thermobifida fusca catalase with peroxidase activity

    NARCIS (Netherlands)

    Lončar, Nikola; Fraaije, Marco W

    2015-01-01

    Thermobifida fusca is a mesothermophilic organism known for its ability to degrade plant biomass and other organics, and it was demonstrated that it represents a rich resource of genes encoding for potent enzymes for biocatalysis. The thermostable catalase from T. fusca has been cloned and overexpre

  16. Structural and Functional Characterizations of SsgB, a Conserved Activator of Developmental Cell Division in Morphologically Complex Actinomycetes

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    Xu, Qingping; Traag, Bjørn A.; Willemse, Joost; McMullan, Daniel; Miller, Mitchell D.; Elsliger, Marc-André; Abdubek, Polat; Astakhova, Tamara; Axelrod, Herbert L.; Bakolitsa, Constantina; Carlton, Dennis; Chen, Connie; Chiu, Hsiu-Ju; Chruszcz, Maksymilian; Clayton, Thomas; Das, Debanu; Deller, Marc C.; Duan, Lian; Ellrott, Kyle; Ernst, Dustin; Farr, Carol L.; Feuerhelm, Julie; Grant, Joanna C.; Grzechnik, Anna; Grzechnik, Slawomir K.; Han, Gye Won; Jaroszewski, Lukasz; Jin, Kevin K.; Klock, Heath E.; Knuth, Mark W.; Kozbial, Piotr; Krishna, S. Sri; Kumar, Abhinav; Marciano, David; Minor, Wladek; Mommaas, A. Mieke; Morse, Andrew T.; Nigoghossian, Edward; Nopakun, Amanda; Okach, Linda; Oommachen, Silvya; Paulsen, Jessica; Puckett, Christina; Reyes, Ron; Rife, Christopher L.; Sefcovic, Natasha; Tien, Henry J.; Trame, Christine B.; van den Bedem, Henry; Wang, Shuren; Weekes, Dana; Hodgson, Keith O.; Wooley, John; Deacon, Ashley M.; Godzik, Adam; Lesley, Scott A.; Wilson, Ian A.; van Wezel, Gilles P.; (Leiden-MC); (SLAC); (Scripps); (UV); (UCSD); (Burnham)

    2010-01-20

    SsgA-like proteins (SALPs) are a family of homologous cell division-related proteins that occur exclusively in morphologically complex actinomycetes. We show that SsgB, a subfamily of SALPs, is the archetypal SALP that is functionally conserved in all sporulating actinomycetes. Sporulation-specific cell division of Streptomyces coelicolor ssgB mutants is restored by introduction of distant ssgB orthologues from other actinomycetes. Interestingly, the number of septa (and spores) of the complemented null mutants is dictated by the specific ssgB orthologue that is expressed. The crystal structure of the SsgB from Thermobifida fusca was determined at 2.6 {angstrom} resolution and represents the first structure for this family. The structure revealed similarities to a class of eukaryotic 'whirly' single-stranded DNA/RNA-binding proteins. However, the electro-negative surface of the SALPs suggests that neither SsgB nor any of the other SALPs are likely to interact with nucleotide substrates. Instead, we show that a conserved hydrophobic surface is likely to be important for SALP function and suggest that proteins are the likely binding partners.

  17. EXTRACELLULAR CELLULOLYTIC COMPLEXES PRODUCTION BY MICROSCOPIC FUNGI

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    S. O. Syrchin

    2015-10-01

    Full Text Available The aim of this work was to screen and to study the effect of inducers on the synthesis of the cellulolytic enzyme complexes by microscopic fungi. Cellulolytic and xylanolytic activities were determined by reducing sugar with DNS reagent, and β-glucosidase activity by pNPG hydrolysis. The enzyme preparations were obtained by ammonium sulphate precipitation. Among 32 studied strains of microscopic fungi 14 produced cellulo- and xylanolytic enzyme complexes. Fusarium sp. 5 and Fennellia sp. 2806 demonstrated the highest levels of all studied enzyme activities. Enzyme preparations with high endo-, exoglucanase, xylanase and β-glucosidase activities were obtained from these strains. Fusarium sp. 5 and Fennellia sp. 2806 were active producers of cellulase enzyme complexes during growth on natural substrates. It was shown that inductors of cellulolytic enzymes in Fusarium sp. 5 and Fennellia sp. 2806 differed from the ones in Trichoderma reesei.

  18. Natural Products from Mangrove Actinomycetes

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    Dong-Bo Xu

    2014-05-01

    Full Text Available Mangroves are woody plants located in tropical and subtropical intertidal coastal regions. The mangrove ecosystem is becoming a hot spot for natural product discovery and bioactivity survey. Diverse mangrove actinomycetes as promising and productive sources are worth being explored and uncovered. At the time of writing, we report 73 novel compounds and 49 known compounds isolated from mangrove actinomycetes including alkaloids, benzene derivatives, cyclopentenone derivatives, dilactones, macrolides, 2-pyranones and sesquiterpenes. Attractive structures such as salinosporamides, xiamycins and novel indolocarbazoles are highlighted. Many exciting compounds have been proven as potential new antibiotics, antitumor and antiviral agents, anti-fibrotic agents and antioxidants. Furthermore, some of their biosynthetic pathways have also been revealed. This review is an attempt to consolidate and summarize the past and the latest studies on mangrove actinomycetes natural product discovery and to draw attention to their immense potential as novel and bioactive compounds for marine drugs discovery.

  19. Cloning and recombinant expression of a cellulase from the cellulolytic strain Streptomyces sp. G12 isolated from compost

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    Amore Antonella

    2012-12-01

    Full Text Available Abstract Background The use of lignocellulosic materials for second generation ethanol production would give several advantages such as minimizing the conflict between land use for food and fuel production, providing less expensive raw materials than conventional agricultural feedstock, allowing lower greenhouse gas emissions than those of first generation ethanol. However, cellulosic biofuels are not produced at a competitive level yet, mainly because of the high production costs of the cellulolytic enzymes. Therefore, this study was aimed at discovering new cellulolytic microorganisms and enzymes. Results Different bacteria isolated from raw composting materials obtained from vegetable processing industry wastes were screened for their cellulolytic activity on solid medium containing carboxymethylcellulose. Four strains belonging to the actinomycetes group were selected on the basis of their phenotypic traits and cellulolytic activity on solid medium containing carboxymethylcellulose. The strain showing the highest cellulolytic activity was identified by 16S rRNA sequencing as belonging to Streptomyces genus and it was designated as Streptomyces sp. strain G12. Investigating the enzymes responsible for cellulase activity produced by Streptomyces G12 by proteomic analyses, two endoglucanases were identified. Gene coding for one of these enzymes, named CelStrep, was cloned and sequenced. Molecular analysis showed that the celstrep gene has an open reading frame encoding a protein of 379 amino acid residues, including a signal peptide of 37 amino acid residues. Comparison of deduced aminoacidic sequence to the other cellulases indicated that the enzyme CelStrep can be classified as a family 12 glycoside hydrolase. Heterologous recombinant expression of CelStrep was carried out in Escherichia coli, and the active recombinant enzyme was purified from culture supernatant and characterized. It catalyzes the hydrolysis of carboxymethylcellulose

  20. IDENTIFICATION AND CHARACTERIZATION OF THERMOBIFIDA FUSCA GENES INVOLVED IN PLANT CELL WALL DEGRADATION.

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    David B. Wilson

    2006-01-23

    Micro-array experiments identified a number of Thermobifida fusca genes which were upregulated by growth on cellulose or plant biomass. Five of these genes were cloned, overexpressed in E. coli and the expressed proteins were purified and characterized. These were a xyloglucanase,a 1-3,beta glucanase, a family 18 hydrolase and twocellulose binding proteins that contained no catalytic domains. The catalyic domain of the family 74 endoxyloglucanase with a C-terminal, cellulose binding module was crystalized and its 3-dimensional structure was determined by X-ray crystallography.

  1. Discovery of novel metabolites from marine actinomycetes.

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    Lam, Kin S

    2006-06-01

    Recent findings from culture-dependent and culture-independent methods have demonstrated that indigenous marine actinomycetes exist in the oceans and are widely distributed in different marine ecosystems. There is tremendous diversity and novelty among the marine actinomycetes present in marine environments. Progress has been made to isolate novel actinomycetes from samples collected at different marine environments and habitats. These marine actinomycetes produce different types of new secondary metabolites. Many of these metabolites possess biological activities and have the potential to be developed as therapeutic agents. Marine actinomycetes are a prolific but underexploited source for the discovery of novel secondary metabolites.

  2. Characterization of cytochrome P450 monooxygenase CYP154H1 from the thermophilic soil bacterium Thermobifida fusca

    NARCIS (Netherlands)

    Schallmey, Anett; den Besten, Gijs; Teune, Ite G. P.; Kembaren, Roga F.; Janssen, Dick B.

    2011-01-01

    Cytochrome P450 monooxygenases are valuable biocatalysts due to their ability to hydroxylate unactivated carbon atoms using molecular oxygen. We have cloned the gene for a new cytochrome P450 monooxygenase, named CYP154H1, from the moderately thermophilic soil bacterium Thermobifida fusca. The enzym

  3. Actinomycete integrative and conjugative elements

    NARCIS (Netherlands)

    Poele, Evelien M. te; Bolhuis, Henk; Dijkhuizen, Lubbert

    2008-01-01

    This paper reviews current knowledge on actinomycete integrative and conjugative elements (AICEs). The best characterised AICEs, pSAM2 of Streptomyces ambofaciens (10.9 kb), SLP1 (17.3 kb) of Streptomyces coelicolor and pMEA300 of Amycolatopsis methanolica (13.3 kb), are present as integrative eleme

  4. A New Esterase from Thermobifida halotolerans Hydrolyses Polyethylene Terephthalate (PET and Polylactic Acid (PLA

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    Georg Steinkellner

    2012-02-01

    Full Text Available A new esterase from Thermobifida halotolerans (Thh_Est was cloned and expressed in E. coli and investigated for surface hydrolysis of polylactic acid (PLA and polyethylene terephthalate (PET. Thh_Est is a member of the serine hydrolases superfamily containing the -GxSxG- motif with 85–87% homology to an esterase from T. alba, to an acetylxylan esterase from T. fusca and to various Thermobifida cutinases. Thh_Est hydrolyzed the PET model substrate bis(benzoyloxyethylterephthalate and PET releasing terephthalic acid and mono-(2-hydroxyethyl terephthalate in comparable amounts (19.8 and 21.5 mmol/mol of enzyme while no higher oligomers like bis-(2-hydroxyethyl terephthalate were detected. Similarly, PLA was hydrolyzed as indicated by the release of lactic acid. Enzymatic surface hydrolysis of PET and PLA led to a strong hydrophilicity increase, as quantified with a WCA decrease from 90.8° and 75.5° to 50.4° and to a complete spread of the water drop on the surface, respectively.

  5. NOVEL BIOACTIVE COMPOUNDS FROM MANGROVE DERIVED ACTINOMYCETES

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    Kumari Amrita

    2012-09-01

    Full Text Available Mangrove is most productive and unexplored ecosystem that approximately covers one fourth of world coastline with high diversity of thriving organism. Recently the rate of isolation of novel bioactive compounds from microorganism living in mangrove forest has tremendously increased which is reflected in significant hasten for exploration of mangrove actinomycetes. Actinomycetes are group of bacteria which are extremely interesting as active producers of many primary and secondary metabolites. Many survey reports has depicted that the biologically active compounds which have been obtained so far from microbes, 45 percent are produced by actinomycetes, 38 percent by fungi and 17 percent by unicellular bacteria. Actinomycetes from mangrove environment provide diverse and are potential rich source of antibiotics, anticancer, antifungal and antiviral agent, enzyme and enzyme inhibitor. Mangrove actinomycetes are a prolific but underexploited source for the discovery of novel secondary metabolites.

  6. Screening genus Penicillium for producers of cellulolytic and xylanolytic enzymes

    DEFF Research Database (Denmark)

    Krogh, Kristian Bertel Rømer; Mørkeberg, Astrid; Frisvad, Jens Christian;

    2004-01-01

    For enzymatic hydrolysis of lignocellulosic material, cellulolytic enzymes from Trichoderma reesei are most commenly used, but, there is a need for more efficient enzyme cocktails. In this study, the production of cellulolytic and xylanolytic enzymes was investigated in 12 filamentous fungi from...

  7. Comparative Genomics of Core Metabolism Genes of Cellulolytic and Non-cellulolytic Clostridium Species.

    Science.gov (United States)

    Lal, Sadhana; Levin, David B

    Microbial production of fuels such as ethanol, butanol, hydrogen (H2), and methane (CH4) from waste biomass has the potential to provide sustainable energy systems that can displace fossil fuel consumption. Screening for microbial diversity and genome sequencing of a wide-range of microorganisms can identify organisms with natural abilities to synthesize these alternative fuels and/or other biotechnological applications. Clostridium species are the most widely studied strict anaerobes capable of fermentative synthesis of ethanol, butanol, or hydrogen directly from waste biomass. Clostridium termitidis CT1112 is a mesophilic, cellulolytic species capable of direct cellulose fermentation to ethanol and organic acids, with concomitant synthesis of H2 and CO2. On the basis of 16S ribosomal RNA (rRNA) and chaperonin 60 (cpn60) gene sequence data, phylogenetic analyses revealed a close relationship between C. termitidis and C. cellobioparum. Comparative bioinformatic analyses of the C. termitidis genome with 18 cellulolytic and 10 non-cellulolytic Clostridium species confirmed this relationship, and further revealed that the majority of core metabolic pathway genes in C. termitidis and C. cellobioparum share more than 90% amino acid sequence identity. The gene loci and corresponding amino acid sequences of the encoded enzymes for each pathway were correlated by percentage identity, higher score (better alignment), and lowest e-value (most significant "hit"). In addition, the function of each enzyme was proposed by conserved domain analysis. In this chapter we discuss the comparative analysis of metabolic pathways involved in synthesis of various useful products by cellulolytic and non-cellulolytic biofuel and solvent producing Clostridium species. This study has generated valuable information concerning the core metabolism genes and pathways of C. termitidis CT1112, which is helpful in developing metabolic engineering strategies to enhance its natural capacity for better

  8. Therapeutically Active Biomolecules from Marine Actinomycetes

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    Mani Jayaprakashvel

    2012-09-01

    Full Text Available For the past few centuries, the biological sources of terrestrial origin have been explored and exploited for bioactive metabolites. This has resulted in the stagnancy of discovering either novel compounds or compounds with novel bioactivities. Thus, researchers across the globe have started exploring our big Oceans, for the search of bioactive metabolites. During the past few decades, the research on bioactive metabolites from marine biological resources has geared up and among the sources marine actinomycetes are proved to be best. Marine actinomycetes, the filamentous bacteria from marine environment have been intensively studied for bioactive metabolites. The biological diversity of marine actinomycetes was found to be enormous, thanks to culture dependent and culture independent biodiversity approaches. This great diversity of marine actinomycetes has offered greater chemical diversity. The diverse chemical compounds of marine actinomycetes have been found to have various biological activities such as antimicrobial, anti-tumor, anti-malarial, anti-algal, antioxidant, anti-inflammatory etc. These various bioactive metabolites of marine actinomycetes are having scope for developing as potent therapeutic agents. The potential of marine actinomycetes is rightly realized though the current biological wealth of these organisms isrelatively unexplored.

  9. An Efficient and Improved Methodology for the Screening of Industrially Valuable Xylano-Pectino-Cellulolytic Microbes

    OpenAIRE

    2015-01-01

    Xylano-pectino-cellulolytic enzymes are valuable enzymes of the industrial sector. In our earlier study, we have reported a novel and cost effective methodology for the qualitative screening of cellulase-free xylano-pectinolytic microorganisms by replacing the commercial, highly expensive substrates with agricultural residues, but the microorganisms with xylanolytic, pectinolytic, cellulolytic, xylano-pectinolytic, xylano-cellulolytic, pectino-cellulolytic, and xylano-pectino-cellulolytic pot...

  10. Antitumor Compounds from Marine Actinomycetes

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    José A. Salas

    2009-06-01

    Full Text Available Chemotherapy is one of the main treatments used to combat cancer. A great number of antitumor compounds are natural products or their derivatives, mainly produced by microorganisms. In particular, actinomycetes are the producers of a large number of natural products with different biological activities, including antitumor properties. These antitumor compounds belong to several structural classes such as anthracyclines, enediynes, indolocarbazoles, isoprenoides, macrolides, non-ribosomal peptides and others, and they exert antitumor activity by inducing apoptosis through DNA cleavage mediated by topoisomerase I or II inhibition, mitochondria permeabilization, inhibition of key enzymes involved in signal transduction like proteases, or cellular metabolism and in some cases by inhibiting tumor-induced angiogenesis. Marine organisms have attracted special attention in the last years for their ability to produce interesting pharmacological lead compounds.

  11. Detection and identification of novel actinomycetes.

    Science.gov (United States)

    Williams, S T; Locci, R; Beswick, A; Kurtböke, D I; Kuznetsov, V D; Le Monnier, F J; Long, P F; Maycroft, K A; Palma, R A; Petrolini, B

    1993-10-01

    The actinomycetes are well known as a group of filamentous, Gram-positive bacteria that produce many useful secondary metabolites, including antibiotics and enzymes. Although they have been intensively studied for both theoretical and practical objectives, there is much scope for developing our basic knowledge of the means of detection and isolation of these microbes. This session concentrated on new methods for the detection and identification of novel actinomycetes from a range of environments. Approaches to the detection of actinomycetes ranged from investigations of neglected habitats and extreme environments (e.g. alkaline soils and oil drills) to the analysis of DNA extracted from the environment and use of specific phages. The continuing problems of the identification of actinomycete isolates were also considered. Topics discussed included use of phage typing, DNA probes, and correlation between phenetic and genotypic species of Streptomyces.

  12. Deep Sea Actinomycetes and Their Secondary Metabolites

    Directory of Open Access Journals (Sweden)

    Kui Hong

    2017-05-01

    Full Text Available Deep sea is a unique and extreme environment. It is a hot spot for hunting marine actinomycetes resources and secondary metabolites. The novel deep sea actinomycete species reported from 2006 to 2016 including 21 species under 13 genera with the maximum number from Microbacterium, followed by Dermacoccus, Streptomyces and Verrucosispora, and one novel species for the other 9 genera. Eight genera of actinomycetes were reported to produce secondary metabolites, among which Streptomyces is the richest producer. Most of the compounds produced by the deep sea actinomycetes presented antimicrobial and anti-cancer cell activities. Gene clusters related to biosynthesis of desotamide, heronamide, and lobophorin have been identified from the deep sea derived Streptomyces.

  13. Elicitation of secondary metabolism in actinomycetes.

    Science.gov (United States)

    Abdelmohsen, Usama Ramadan; Grkovic, Tanja; Balasubramanian, Srikkanth; Kamel, Mohamed Salah; Quinn, Ronald J; Hentschel, Ute

    2015-11-01

    Genomic sequence data have revealed the presence of a large fraction of putatively silent biosynthetic gene clusters in the genomes of actinomycetes that encode for secondary metabolites, which are not detected under standard fermentation conditions. This review focuses on the effects of biological (co-cultivation), chemical, as well as molecular elicitation on secondary metabolism in actinomycetes. Our review covers the literature until June 2014 and exemplifies the diversity of natural products that have been recovered by such approaches from the phylum Actinobacteria.

  14. Not so monofunctional--a case of thermostable Thermobifida fusca catalase with peroxidase activity.

    Science.gov (United States)

    Lončar, Nikola; Fraaije, Marco W

    2015-03-01

    Thermobifida fusca is a mesothermophilic organism known for its ability to degrade plant biomass and other organics, and it was demonstrated that it represents a rich resource of genes encoding for potent enzymes for biocatalysis. The thermostable catalase from T. fusca has been cloned and overexpressed in Escherichia coli with a yield of 400 mg/L. Heat treatment of disrupted cells at 60 °C for 1 h resulted in enzyme preparation of high purity; hence, no chromatography steps are needed for large-scale production. Except for catalyzing the dismutation of hydrogen peroxide, TfuCat was also found to catalyze oxidations of phenolic compounds. The catalase activity was comparable to other described catalases while peroxidase activity was quite remarkable with a k obs of nearly 1000 s(-1) for catechol. Site directed mutagenesis was used to alter the ratio of peroxidase/catalase activity. Resistance to inhibition by classic catalase inhibitors and an apparent melting temperature of 74 °C classifies this enzyme as a robust biocatalyst. As such, it could compete with other commercially available catalases while the relatively high peroxidase activity also offers new biocatalytic possibilities.

  15. Heterologous expression and biochemical characterization of glucose isomerase from Thermobifida fusca.

    Science.gov (United States)

    Deng, Hui; Chen, Sheng; Wu, Dan; Chen, Jian; Wu, Jing

    2014-06-01

    Glucose isomerase (GIase) catalyzes the isomerization of D-glucose to D-fructose. The GIase from Thermobifida fusca WSH03-11 was expressed in Escherichia coli BL21(DE3), and the purified enzyme took the form of a tetramer in solution and displayed a pI value of 5.05. The temperature optimum of GIase was 80 °C and its half life was about 2 h at 80 °C or 15 h at 70 °C. The pH optimum of GIase was 10 and the enzyme retained 95 % activity over the pH range of 5-10 after incubating at 4 °C for 24 h. Kinetic studies showed that the K m and K cat values of the enzyme are 197 mM and 1,688 min(-1), respectively. The maximum conversion yield of glucose (45 %, w/v) to fructose of the enzyme was 53 % at pH 7.5 and 70 °C. The present study provides the basis for the industrial application of recombinant T. fusca GIase in the production of high fructose syrup.

  16. Cellulase production from treated oil palm empty fruit bunch degradation by locally isolated Thermobifida fusca

    Directory of Open Access Journals (Sweden)

    M. Nazli Naim

    2013-02-01

    Full Text Available The aim of this research was to evaluate the production of cellulases from locally isolated bacteria, Thermobifida fusca, using thermal and chemical treated oil palm empty fruit bunch (OPEFB as substrate in liquid-state fermentation (LSF. T. fusca was successfully isolated and was a dominant cellulase producer in OPEFB composting at the thermophilic stage. Analysis of the surface morphology of OPEFB samples using Scanning Electron Microscopy (SEM showed that the most significant changes after the combination of thermal and chemical pretreatment was the removal of silica bodies, and this observation was supported by X-ray Diffraction analysis (XRD, Fourier Transform Infrared (FTIR, and Thermogravimetric analysis (TG showing changes on the hemicelluloses, cellulose, and lignin structures throughout the pretreatment process. As a result of the pretreatment, higher cellulase production by T. fusca was obtained. The highest activity for CMCase, FPase, and β-glucosidase using optimally treated OPEFB were 0.24 U/mL, 0.34 U/mL, and 0.04 U/mL, respectively. Therefore, it can be suggested that the combination of chemical and thermal pretreatments enhances the degradation of OPEFB for subsequent use as fermentation substrate, contributing to a higher cellulases yield by T. fusca.

  17. Cloning, Expression and Identification of a New Trehalose Synthase Gene from Thermobifida fusca Genome

    Institute of Scientific and Technical Information of China (English)

    Yu-Tuo WEI; Ri-Bo HUANG; Qi-Xia ZHU; Zhao-Fei LUO; Fu-Shen LU; Fa-Zhong CHEN; Qing-Yan WANG; Kun HUANG; Jian-Zhong MENG; Rong WANG

    2004-01-01

    A new open reading frame in Thermobifida fusca sequenced genome was identified to encode a new trehalose synthase, annotated as "glycosidase" in the GenBank database, by bioinformatics searching and experimental validation. The gene had a length of 1830 bp with about 65% GC content and encoded for a new trehalose synthase with 610 amino acids and deduced molecular weight of 66 kD. The high GC content seemed not to affect its good expression in E. coli BL21 in which the target protein could account for as high as 15% of the total cell proteins. The recombinant enzyme showed its optimal activities at 25 ℃ and pH 6.5 when it converted substrate maltose into trehalose. However it would divert a high proportion of its substrate into glucose when the temperature was increased to 37 ℃, or when the enzyme concentration was high Its activity was not inhibited by 5 mM heavy metals such as Cu2+, Mn2+, and Zn2+ but affected by high concentration of glucose. Blasting against the database indicated that amino acid sequence of this protein had maximal 69% homology with the known trehalose synthases, and two highly conserved segments of the protein sequence were identified and their possible linkage with functions was discussed.

  18. Production of cellulolytic enzymes from ascomycetes

    DEFF Research Database (Denmark)

    Hansen, Gustav Hammerich; Lübeck, Mette; Frisvad, Jens Christian;

    2015-01-01

    Optimizing production of cellulose degrading enzymes is of great interest in order to increase the feasibility of constructing biorefinery facilities for a sustainable supply of energy and chemical products. The ascomycete phylum has a large potential for the production of cellulolytic enzymes....... Although numerous enzymatic profiles have already been unraveled, the research has been covering only a limited number of species and genera, thus leaving many ascomycetes to be analyzed. Such analysis requires choosing appropriate media and cultivation methods that ensure enzyme profiles with high...... specificities and activities. However, the choice of media, cultivation methods and enzyme assays highly affect the enzyme activity profile observed. This review provides an overview of enzymatic profiles for several ascomycetes covering phylogenetically distinct genera and species. The profiles of cellulose...

  19. Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Yu; Tang, Lan; Henriksen, Svend Hostgaard Bang

    2016-05-17

    The present invention provides isolated polypeptides having cellulolytic enhancing activity and isolated polynucleotides encoding the polypeptides. The invention also provides nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

  20. Exploring the biocatalytic potential of a DyP-type peroxidase by profiling the substrate acceptance of Thermobifida fusca DyP peroxidase

    NARCIS (Netherlands)

    Loncar, Nikola; Colpa, Dana I.; Fraaije, Marco W.

    2016-01-01

    Dye-decolorizing peroxidases (DyPs) represent a new class of oxidative enzymes for which the natural substrates are largely unknown. To explore the biocatalytic potential of a DyP, we have studied the substrate acceptance profile of a robust DyP peroxidase, a DyP from Thermobifida fusca (TfuDyP). Wh

  1. Cloning, expression and characterization of a family-74 xyloglucanase from Thermobifida fusca.

    Science.gov (United States)

    Irwin, Diana C; Cheng, Mark; Xiang, Bosong; Rose, Jocelyn K C; Wilson, David B

    2003-07-01

    Thermobifida fusca xyloglucan-specific endo-beta-1,4-glucanase (Xeg)74 and the Xeg74 catalytic domain (CD) were cloned, expressed in Escherichia coli, purified and characterized. This enzyme has a glycohydrolase family-74 CD that is a specific xyloglucanase followed by a family-2 carbohydrate binding module at the C terminus. The Michaelis constant (Km) and maximal rate (Vmax) values for hydrolysis of tamarind seed xyloglucan (tamXG) are 2.4 micro m and 966 micro mol xyloglucan oligosaccharides (XGOs) min-1. micro mol protein-1. More than 75% of the activity was retained after a 16-h incubation at temperatures up to 60 degrees C. The enzyme was most active at pH 6.0-9.4. NMR analysis showed that its catalytic mechanism is inverting. The oligosaccharide products from hydrolysis of tamXG were determined by MS analysis. Cel9B, an active carboxymethylcellulose (CMC)ase from T. fusca, was also found to have activity on xyloglucan (XG) at 49 micro mol.min-1. micro mol protein-1, but it could not hydrolyze XG units containing galactose. An XG/cellulose composite was prepared by growing Gluconacetobacterxylinus on glucose with tamXG in the medium. Although a mixture of purified cellulases was unable to degrade this material, the composite material was fully hydrolyzed when Xeg74 was added. T. fusca was not able to grow on tamXG, but Xeg74 was found in the culture supernatant at the same level as was found in cultures grown on Solka Floc. The function of this enzyme appears to be to break down the XG surrounding cellulose fibrils found in biomass so that T. fusca can utilize the cellulose as a carbon source.

  2. Surface engineering of a cutinase from Thermobifida cellulosilytica for improved polyester hydrolysis.

    Science.gov (United States)

    Herrero Acero, Enrique; Ribitsch, Doris; Dellacher, Anita; Zitzenbacher, Sabine; Marold, Annemarie; Steinkellner, Georg; Gruber, Karl; Schwab, Helmut; Guebitz, Georg M

    2013-10-01

    Modeling and comparison of the structures of the two closely related cutinases Thc_Cut1 and Thc_Cut2 from Thermobifida cellulosilytica DSM44535 revealed that dissimilarities in their electrostatic and hydrophobic surface properties in the vicinity to the active site could be responsible for pronounced differences in hydrolysis efficiencies of polyester (i.e., PET, polyethyleneterephthalate). To investigate this hypothesis in more detail, selected amino acids of surface regions outside the active site of Thc_Cut2, which hydrolyzes PET much less efficiently than Thc_Cut1 were exchanged by site-directed mutagenesis. The mutants were expressed in E. coli BL21-Gold(DE3), purified and characterized regarding their specific activities and kinetic parameters on soluble substrates and their ability to hydrolyze PET and the PET model substrate bis(benzoyloxyethyl) terephthalate (3PET). Compared to Thc_Cut2, mutants carrying Arg29Asn and/or Ala30Val exchanges showed considerable higher specific activity and higher kcat /KM values on soluble substrates. Exchange of the positively charged arginine (Arg19 and Arg29) located on the enzyme surface to the non-charged amino acids serine and asparagine strongly increased the hydrolysis activity for 3PET and PET. In contrast, exchange of the uncharged glutamine (Glu65) by the negatively charged glutamic acid lead to a complete loss of hydrolysis activity on PET films. These findings clearly demonstrate that surface properties (i.e., amino acids located outside the active site on the protein surface) play an important role in PET hydrolysis.

  3. Actinomycetes and the IUD: an update.

    Science.gov (United States)

    Gupta, P K; Erozan, Y S; Frost, J K

    1978-01-01

    To date, actinomycetes have been identified in 540 vagino-pancervical (Fast) smears from 520 women. In each case, a foreign body has been present, usually an IUD (517 cases). The Dalkon Shield preponderates, perhaps reflecting physician preference at the center. IUD usage duration has varied from 6 weeks to 13 years. Approximately 25% of symptomatic IUD users requesting treatment have cytologic evidence of actinomycetes. The preoperative diagnosis of pelvic actinomycetes was suggested in most cases based on cytologic evidence alone. Immunofluorescent studies have been performed in 266 cases with species-specific antisera, and Actinomyces israelii identified in 250. Protozoal organisms in the Fast smears have been noted in 8 cases (1.5%) and are commonly intimately associated with Actinomyces. It is agreed that the oropharnyx serves as the possible source of lower genital tract Actinomyces infection. In such cases, IUD removal and antibiotic treatment are recommended.

  4. Culturable rare Actinomycetes: diversity, isolation and marine natural product discovery.

    Science.gov (United States)

    Subramani, Ramesh; Aalbersberg, William

    2013-11-01

    Rare Actinomycetes from underexplored marine environments are targeted in drug discovery studies due to the Actinomycetes' potentially huge resource of structurally diverse natural products with unusual biological activity. Of all marine bacteria, 10 % are Actinomycetes, which have proven an outstanding and fascinating resource for new and potent bioactive molecules. Past and present efforts in the isolation of rare Actinomycetes from underexplored diverse natural habitats have resulted in the isolation of about 220 rare Actinomycete genera of which more than 50 taxa have been reported to be the producers of 2,500 bioactive compounds. That amount represents greater than 25 % of the total Actinomycetes metabolites, demonstrating that selective isolation methods are being developed and extensively applied. Due to the high rediscovery rate of known compounds from Actinomycetes, a renewed interest in the development of new antimicrobial agents from rare and novel Actinomycetes is urgently required to combat the increasing number of multidrug-resistant human pathogens. To facilitate that discovery, this review updates all selective isolation media including pretreatment and enrichment methods for the isolation of marine rare Actinomycetes. In addition, this review demonstrates that discovering new compounds with novel scaffolds can be increased by intensive efforts in isolating and screening rare marine genera of Actinomycetes. Between 2007 and mid-2013, 80 new rare Actinomycete species were reported from marine habitats. They belong to 23 rare families, of which three are novel, and 20 novel genera. Of them, the family Micromonosporaceae is dominant as a producer of promising chemical diversity.

  5. Systematic and biotechnological aspects of halophilic and halotolerant actinomycetes.

    Science.gov (United States)

    Hamedi, Javad; Mohammadipanah, Fatemeh; Ventosa, Antonio

    2013-01-01

    More than 70 species of halotolerant and halophilic actinomycetes belonging to at least 24 genera have been validly described. Halophilic actinomycetes are a less explored source of actinomycetes for discovery of novel bioactive secondary metabolites. Degradation of aliphatic and aromatic organic compounds, detoxification of pollutants, production of new enzymes and other metabolites such as antibiotics, compatible solutes and polymers are other potential industrial applications of halophilic and halotolerant actinomycetes. Especially new bioactive secondary metabolites that are derived from only a small fraction of the investigated halophilic actinomycetes, mainly from marine habitats, have revealed the huge capacity of this physiological group in production of new bioactive chemical entities. Combined high metabolic capacities of actinomycetes and unique features related to extremophilic nature of the halophilic actinomycetes have conferred on them an influential role for future biotechnological applications.

  6. Cellulolytic potential of thermophilic species from four fungal orders

    DEFF Research Database (Denmark)

    Busk, Peter Kamp; Lange, Lene

    2013-01-01

    . Thermophilic fungi are the only described eukaryotes that can grow at temperatures above 45 ºC. All 16 fungi were able to grow on crystalline cellulose but their secreted enzymes showed widely different cellulolytic activities, pH optima and thermostabilities. Interestingly, in contrast to previous reports, we...... found that some fungi such as Melanocarpus albomyces readily grew on crystalline cellulose and produced cellulases. These results indicate that there are large differences in the cellulolytic potential of different isolates of the same species. Furthermore, all the selected species were able to degrade...... cellulose but the differences in cellulolytic potential and thermostability of the secretome did not correlate to the taxonomic position. PCR amplification and sequencing of 22 cellulase genes from the fungi showed that the level of thermostability of the cellulose-degrading activity could not be inferred...

  7. Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Yu; Duan, Junxin; Tang, Lan; Wu, Wenping

    2016-06-14

    The present invention relates to isolated polypeptides having cellulolytic enhancing activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

  8. Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same

    Energy Technology Data Exchange (ETDEWEB)

    Schnorr, Kirk; Kramer, Randall

    2017-08-08

    The present invention relates to isolated polypeptides having cellulolytic enhancing activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

  9. Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same

    Energy Technology Data Exchange (ETDEWEB)

    Tang, Lan; Liu, Ye; Duan, Junxin; Wu, Wenping; Kramer, Randall

    2017-09-19

    The present invention relates to isolated polypeptides having cellulolytic enhancing activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

  10. Polypeptides having cellulolytic enhancing activity and nucleic acids encoding same

    Energy Technology Data Exchange (ETDEWEB)

    Brown, Kimberly; Harris, Paul; Zaretsky, Elizabeth; Re, Edward; Vlasenko, Elena; McFarland, Keith; Lopez de Leon, Alfredo

    2017-09-05

    The present invention relates to isolated polypeptides having cellulolytic enhancing activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods for producing and using the polypeptides.

  11. Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same

    Energy Technology Data Exchange (ETDEWEB)

    Schnorr, Kirk; Kramer, Randall

    2016-08-09

    The present invention relates to isolated polypeptides having cellulolytic enhancing activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

  12. Polypeptides having cellulolytic enhancing activity and nucleic acids encoding same

    Energy Technology Data Exchange (ETDEWEB)

    Brown, Kimberly; Harris, Paul; Zaretsky, Elizabeth; Re, Edward; Vlasenko, Elena; McFarland, Keith; Lopez de Leon, Alfredo

    2016-08-09

    The present invention relates to isolated polypeptides having cellulolytic enhancing activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods for producing and using the polypeptides.

  13. Polypeptides having cellulolytic enhancing activity and nucleic acids encoding same

    Energy Technology Data Exchange (ETDEWEB)

    Brown, Kimberly; Harris, Paul; Zaretsky, Elizabeth; Re, Edward; Vlasenko, Elena; McFarland, Keith; Lopez de Leon, Alfredo

    2014-09-30

    The present invention relates to isolated polypeptides having cellulolytic enhancing activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods for producing and using the polypeptides.

  14. Distribution of actinomycetes in near-shore tropical marine sediments.

    OpenAIRE

    Jensen, P.R.; Dwight, R; Fenical, W.

    1991-01-01

    Actinomycetes were isolated from near-shore marine sediments collected at 15 island locations throughout the Bahamas. A total of 289 actinomycete colonies were observed, and all but 6 could be assigned to the suprageneric groups actinoplanetes and streptomycetes. A bimodal distribution in the actinomycete population in relation to depth was recorded, with the maximum numbers occurring in the shallow and deep sampling sites. This distribution can be accounted for by a rapid decrease in strepto...

  15. Field studies on two rock phosphate solubilizing actinomycete isolates as biofertilizer sources

    Science.gov (United States)

    Mba, Caroline C.

    1994-03-01

    Recently biotechnology is focusing attention on utilization of biological resources to solve a number of environmental problems such as soil fertility management. Results of microbial studies on earthworm compost in the University of Nigeria farm identified a number of rock phosphate solubilizing actinomycetes. Two of these, isclates 02 and 13, were found to be efficient rock phosphate (RP) solubilizers and fast-growing cellulolytic microbes producing extracellular hydrolase enzymes. In this preliminary field study the two microbial isolates were investigated with respect to their effects on the growth of soybean and egusi as well as their effect on the incidence of toxicity of poultry droppings. Application of these isolates in poultry manure-treated field plots, as microbial fertilizers, brought about yield increases of 43% and 17% with soybeans and 19% and 33% with egusi, respectively. Soil properties were also improved. With isolates 02 and 13, the soil available phosphorus increased at the five-leaf stage, while N-fixation in the soil increased by 45% or 11% relative to control. It was further observed that air-dried poultry manure after four days of incubation was still toxic to soybean. The toxic effect of the applied poultry manure was reduced or eliminated with microbial fertilizers 02 or 13, respectively. The beneficial effects of the microbial organic fertilizer are discussed. Justification for more intensive research on rock phosphate organic fertilizer is highlighted.

  16. Marine actinomycetes: an ongoing source of novel bioactive metabolites.

    Science.gov (United States)

    Subramani, Ramesh; Aalbersberg, William

    2012-12-20

    Actinomycetes are virtually unlimited sources of novel compounds with many therapeutic applications and hold a prominent position due to their diversity and proven ability to produce novel bioactive compounds. There are more than 22,000 known microbial secondary metabolites, 70% of which are produced by actinomycetes, 20% from fungi, 7% from Bacillus spp. and 1-2% by other bacteria. Among the actinomycetes, streptomycetes group are considered economically important because out of the approximately more than 10,000 known antibiotics, 50-55% are produced by this genus. The ecological role of actinomycetes in the marine ecosystem is largely neglected and various assumptions meant there was little incentive to isolate marine strains for search and discovery of new drugs. The search for and discovery of rare and new actinomycetes is of significant interest to drug discovery due to a growing need for the development of new and potent therapeutic agents. Modern molecular technologies are adding strength to the target-directed search for detection and isolation of bioactive actinomycetes, and continued development of improved cultivation methods and molecular technologies for accessing the marine environment promises to provide access to this significant new source of chemical diversity with novel/rare actinomycetes including new species of previously reported actinomycetes. Copyright © 2012 Elsevier GmbH. All rights reserved.

  17. Malate dehydrogenases from actinomycetes: structural comparison of Thermoactinomyces enzyme with other actinomycete and Bacillus enzymes.

    OpenAIRE

    1984-01-01

    Malate dehydrogenases from bacteria belonging to the genus Thermoactinomyces are tetrameric, like those from Bacillus spp., and exhibit a high degree of structural homology to Bacillus malate dehydrogenase as judged by immunological cross-reactivity. Malate dehydrogenases from other actinomycetes are dimers and do not cross-react with antibodies to Bacillus malate dehydrogenase.

  18. Cellulolytic potential of thermophilic species from four fungal orders

    DEFF Research Database (Denmark)

    Busk, Peter Kamp; Lange, Lene

    2013-01-01

    Elucidation of fungal biomass degradation is important for understanding the turnover of biological materials in nature and has important implications for industrial biomass conversion. In recent years there has been an increasing interest in elucidating the biological role of thermophilic fungi...... and in characterization of their industrially useful enzymes. In the present study we investigated the cellulolytic potential of 16 thermophilic fungi from the three ascomycete orders Sordariales, Eurotiales and Onygenales and from the zygomycete order Mucorales thus covering all fungal orders that include thermophiles....... Thermophilic fungi are the only described eukaryotes that can grow at temperatures above 45 ºC. All 16 fungi were able to grow on crystalline cellulose but their secreted enzymes showed widely different cellulolytic activities, pH optima and thermostabilities. Interestingly, in contrast to previous reports, we...

  19. Reaction kinetics, molecular action, and mechanisms of cellulolytic proteins.

    Science.gov (United States)

    Mosier, N S; Hall, P; Ladisch, C M; Ladisch, M R

    1999-01-01

    Cellulolytic proteins form a complex of enzymes that work together to depolymerize cellulose to the soluble products cellobiose and glucose. Fundamental studies on their molecular mechanisms have been facilitated by advances in molecular biology. These studies have shown homology between cellulases from different microorganisms, and common mechanisms between enzymes whose modes of action have sometimes been viewed as being different, as suggested by the distribution of soluble products. A more complete picture of the cellulolytic action of these proteins has emerged and combines the physical and chemical characteristics of solid cellulose substrates with the specialized structure and function of the cellulases that break it down. This chapter combines the fundamentals of cellulose structure with enzyme function in a manner that relates the cellulose binding and biochemical kinetics at the catalytic site of the proteins to the macroscopic behavior of cellulase enzyme systems.

  20. Characterisation of actinomycetes community from the heavy metals polluted soil

    Directory of Open Access Journals (Sweden)

    Monika Vítězová

    2013-01-01

    Full Text Available The isolation of actinomycetes was performed from soil samples influenced by car-traffic. The acute toxicity of soil leaches was tested by the help of Microtox® bioassay testing system which uses freeze dried luminescent bacteria Photobacterium phosphoreum as the test organisms. The content of heavy metals in biomass of soil microorganisms and in whole soil samples was determinate. 115 strains of actinomycetes were isolated and their total numbers in soil samples were estimated. The acute toxicity of soil influenced the total numbers of actinomycetes. By the help of DNA-DNA reassociation procedure the generic diversity of bacteria was estimated. The identification and differentiation of streptomycetes from the total isolated actinomycetes was made using specific morphological criteria and the gas chromatography-fatty acid methyl ester (GC-FAME analysis. FAME method is adequate only for differentiation of members of genus Streptomyces from other actinomycetes because of their characteristical profile of fatty acids.

  1. Assessment of cellulolytic microorganisms in soils of Nevados Park, Colombia

    Directory of Open Access Journals (Sweden)

    Lizeth Manuela Avellaneda-Torres

    2014-12-01

    Full Text Available A systematized survey was conducted to find soil-borne microbes that degrade cellulose in soils from unique ecosystems, such as the Superpáramo, Páramo, and the High Andean Forest in the Nevados National Natural Park (NNNP, Colombia. These high mountain ecosystems represent extreme environments, such as high levels of solar radiation, low atmospheric pressure, and extreme daily changes in temperature. Cellulolytic activity of the microorganisms was evaluated using qualitative tests, such as growth in selective media followed by staining with congo red and iodine, and quantitative tests to determine the activity of endoglucanase, β-glucosidase, exoglucanase, and total cellulase. Microorganisms were identified using molecular markers, such as the 16S rRNA gene for bacteria and the internal transcribed spacer region (ITS of ribosomal DNA for fungi. Multivariate statistical analysis (MVA was used to select microorganisms with high cellulolytic capacity. A total of 108 microorganisms were isolated from the soils and, in general, the enzymatic activities of fungi were higher than those of bacteria. Our results also found that none of the organisms studied were able to degrade all the components of the cellulose and it is therefore suggested that a combination of bacteria and/or fungi with various enzymatic activities be used to obtain high total cellulolytic activity. This study gives an overview of the potential microorganism that could be used for cellulose degradation in various biotechnological applications and for sustainable agricultural waste treatment.

  2. Characterization of Cellulolytic Bacterial Cultures Grown in Different Substrates

    Directory of Open Access Journals (Sweden)

    Mohamed Idris Alshelmani

    2013-01-01

    Full Text Available Nine aerobic cellulolytic bacterial cultures were obtained from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Culture (DSMZ and the American Type Culture Collection (ATCC. The objectives of this study were to characterize the cellulolytic bacteria and to determine the optimum moisture ratio required for solid state fermentation (SSF of palm kernel cake (PKC. The bacteria cultures were grown on reconstituted nutrient broth, incubated at 30∘C and agitated at 200 rpm. Carboxymethyl cellulase, xylanase, and mannanase activities were determined using different substrates and after SSF of PKC. The SSF was conducted for 4 and 7 days with inoculum size of 10% (v/w on different PKC concentration-to-moisture ratios: 1 : 0.2, 1 : 0.3, 1 : 0.4, and 1 : 0.5. Results showed that Bacillus amyloliquefaciens 1067 DSMZ, Bacillus megaterium 9885 ATCC, Paenibacillus curdlanolyticus 10248 DSMZ, and Paenibacillus polymyxa 842 ATCC produced higher enzyme activities as compared to other bacterial cultures grown on different substrates. The cultures mentioned above also produced higher enzyme activities when they were incubated under SSF using PKC as a substrate in different PKC-to-moisture ratios after 4 days of incubation, indicating that these cellulolytic bacteria can be used to degrade and improve the nutrient quality of PKC.

  3. Sequence-Based Identification of Aerobic Actinomycetes

    Science.gov (United States)

    Patel, Jean Baldus; Wallace, Richard J.; Brown-Elliott, Barbara A.; Taylor, Tony; Imperatrice, Carol; Leonard, Deborah G. B.; Wilson, Rebecca W.; Mann, Linda; Jost, Kenneth C.; Nachamkin, Irving

    2004-01-01

    We investigated the utility of 500-bp 16S rRNA gene sequencing for identifying clinically significant species of aerobic actinomycetes. A total of 28 reference strains and 71 clinical isolates that included members of the genera Streptomyces, Gordonia, and Tsukamurella and 10 taxa of Nocardia were studied. Methods of nonsequencing analyses included growth and biochemical analysis, PCR-restriction enzyme analysis of the 439-bp Telenti fragment of the 65 hsp gene, susceptibility testing, and, for selected isolates, high-performance liquid chromatography. Many of the isolates were included in prior taxonomic studies. Sequencing of Nocardia species revealed that members of the group were generally most closely related to the American Type Culture Collection (ATCC) type strains. However, the sequences of Nocardia transvalensis, N. otitidiscaviarum, and N. nova isolates were highly variable; and it is likely that each of these species contains multiple species. We propose that these three species be designated complexes until they are more taxonomically defined. The sequences of several taxa did not match any recognized species. Among other aerobic actinomycetes, each group most closely resembled the associated reference strain, but with some divergence. The study demonstrates the ability of partial 16S rRNA gene sequencing to identify members of the aerobic actinomycetes, but the study also shows that a high degree of sequence divergence exists within many species and that many taxa within the Nocardia spp. are unnamed at present. A major unresolved issue is the type strain of N. asteroides, as the present one (ATCC 19247), chosen before the availability of molecular analysis, does not represent any of the common taxa associated with clinical nocardiosis. PMID:15184431

  4. Compositions comprising a polypeptide having cellulolytic enhancing activity and a quinone compound and uses thereof

    Energy Technology Data Exchange (ETDEWEB)

    Quinlan, Jason; Xu, Feng; Sweeney, Matthew

    2017-09-05

    The present invention relates to compositions comprising: a polypeptide having cellulolytic enhancing activity and a quinone compound. The present invention also relates to methods of using the compositions.

  5. Compositions comprising a polypeptide having cellulolytic enhancing activity and a bicycle compound and uses thereof

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Feng; Sweeney, Matthew; Quinlan, Jason

    2015-06-16

    The present invention relates to compositions comprising: a polypeptide having cellulolytic enhancing activity and a bicyclic compound. The present invention also relates to methods of using the compositions.

  6. Compositions comprising a polypeptide having cellulolytic enhancing activity and a bicyclic compound and uses thereof

    Energy Technology Data Exchange (ETDEWEB)

    Quinlan, Jason; Xu, Feng; Sweeney, Matthew

    2016-10-04

    The present invention relates to compositions comprising: a polypeptide having cellulolytic enhancing activity and a bicyclic compound. The present invention also relates to methods of using the compositions.

  7. Compositions comprising a polypeptide having cellulolytic enhancing activity and a heterocyclic compound and uses thereof

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Feng; Sweeney, Matthew; Quinlan, Jason

    2016-08-02

    The present invention relates to compositions comprising: a polypeptide having cellulolytic enhancing activity and a heterocyclic compound. The present invention also relates to methods of using the compositions.

  8. Structure and functional analysis of the siderophore periplasmic binding protein from the fuscachelin gene cluster of Thermobifida fusca.

    Science.gov (United States)

    Li, Kunhua; Bruner, Steven D

    2016-01-01

    Iron acquisition is a complex, multicomponent process critical for most organisms' survival and virulence. Small iron chelating molecules, siderophores, mediate transport as key components of common pathways for iron assimilation in many microorganisms. The chemistry and biology of the extraordinary tight and specific metal binding siderophores is of general interest in terms of host/guest chemistry and is a potential target toward the development of therapeutic treatments for microbial virulence. The siderophore pathway of the moderate thermophile, Thermobifida fusca, is an excellent model system to study the process in Gram-positive bacteria. Here we describe the structure and characterization of the siderophore periplasmic binding protein, FscJ from the fuscachelin gene cluster of T. fusca. The structure shows a di-domain arrangement connected with a long α-helix hinge. Several X-ray structures detail ligand-free conformational changes at different pH values, illustrating complex interdomain flexibility of the siderophore receptors. We demonstrated that FscJ has a unique recognition mechanism and details the binding interaction with ferric-fuscachelin A through ITC and docking analysis. The presented work provides a structural basis for the complex molecular mechanisms of siderophore recognition and transportation.

  9. Heterologous expression and biochemical characterization of an endo-β-1,4-glucanase from Thermobifida fusca.

    Science.gov (United States)

    Yan, Peng'an; Su, Lingqia; Chen, Jian; Wu, Jing

    2013-01-01

    The endoglucanase Cel5A from Thermobifida fusca was cloned and expressed in Escherichia coli BL21(DE3). The carboxymethyl cellulase (CMCase) activity in shake flasks and 3-L fermentation scale reached 46.8 and 656.6 IU/mL, respectively. The CMCase activity in 3-L fermentation scale represented the highest yield of T. fusca Cel5A reported so far. Recombinant Cel5A was purified and characterized in detail. The optimum temperature of recombinant enzyme was 80 °C, and the half-life of the enzyme was 132 H at 50 °C and 65 H at 60 °C. The activity of recombinant Cel5A was retained more than 90% over the range of pH 5.0-10.0 with maximal activity at pH 5.5. Using carboxymethyl cellulose as the substrate, the Km and Vmax values were 5.1 mg/mL and 48.7 IU/mg, respectively. The enzyme showed superstability in surfactants and was retained above 90% activity after treatment with sodium dodecyl sulfate, linear alkyl benzene sulfonate, fatty alcohol polyoxyethylene (9) ether, and polyoxyethylene (10) nonyl phenyl ether at 25 °C for 1 H, indicating that the enzyme could be a valuable component in detergents. The potential mechanism of this stability was investigated by analysis of the electrostatic potential of the surface of the enzyme.

  10. [Actinomycetes from mangrove and their secondary metabolites].

    Science.gov (United States)

    Hong, Kui

    2013-11-04

    Mangroves are woody plants located in tropical and subtropical intertidal coastal regions. Driven by the discovery of novel natural products from marine environment, mangrove is becoming a hot spot for actinomycetes resources collection and secondary metabolites (natural products) identification as well as their biosynthesis mechanism investigation. Salinaspora A produced by a Salinispora strain isolated from Bahamas mangrove environment, is in the first clinical trial. Till the time of writing this paper, 24 genera of 11 families and 8 suborders under the actinomycetale have been reported from mangrove, among which 3 are new genera, and 31 are new species. At the same time, secondary metabolites were identified from the mangrove actinomycetes culture, including alkanoids and quinines, azalomycins, antimycins, bezamides and quinazolines, divergolides, indole derivatives, kandenols, macrocyclic dilactones, and the attractive structures, such as the Streptocarbazoles, the multicyclic indolsesquiterpenes, and xiamycin presented unique structures. Their biosynthetic mechanism has also been investigated. Most of the metabolites were isolated from streptomycetes, with a few from Micromonospora and Saccharopolyspora.

  11. Selection and taxonomic identification of carotenoid-producing marine actinomycetes.

    Science.gov (United States)

    Romero, Francisco; Fernández-Chimeno, Rosa Isabel; de la Fuente, Juan Luis; Barredo, José-Luis

    2012-01-01

    Carotenoids are important pigments produced by plants and many microorganisms, including fungi, microalgae, cyanobacteria, and bacteria. Marine actinomycetes are a group of bacteria that produce a variety of metabolites with economic potential. Here, we describe a general method of selecting marine actinomycetes as carotenoids' producers. The screening is carried out at two levels: the first one involves a quick selection of strains by visual color inspection, and the second consists in the analysis of the extracts by HPLC. The taxonomic analysis of the producing strains gives us an overview of the groups of actinomycetes in which carotenoids can be found.

  12. Immobilised cellulolytic and hemicellulolytic enzymes from macrophomina phaseolina

    Energy Technology Data Exchange (ETDEWEB)

    Roy, P.K.; Roy, U.; Dube, D.K.

    1984-01-01

    Cellulolytic enzymes from Macrophomina phaseolina were immobilised in acrylamide polymer. The immobilised enzyme preparation showed activity towards filter paper and cotton. However, the degree of hydrolysis of highly organised cellulose, particularly cotton, appears to be low in comparison with that of soluble substrate. The kinetic studies of immobilised enzymes indicated the presence of diffusional limitations by the increase in Vmax as the particle size decreased. The operational studies suggested that the immobilised enzymes retained the original activities up to 25-29 times in the reuse cycle.

  13. Marine actinomycetes from Madeira Archipelago preliminary taxonomic studies

    National Research Council Canada - National Science Library

    Sara, Rodrigues; Tiago, Dias; Florbela, Pereira; Ilda, Santos Sanches; Susana, Gaudêncio

    2014-01-01

    .... Marine actinomycetes, are a robust resource of chemically prolific novelty. Producing structurally unique biological active secondary metabolites, generating a valuable source for innovative biotechnology and drug discovery[1,2...

  14. Marine actinomycetes: an ongoing source of novel bioactive metabolites

    National Research Council Canada - National Science Library

    Subramani, Ramesh; Aalbersberg, William

    2012-01-01

    ...% are produced by this genus. The ecological role of actinomycetes in the marine ecosystem is largely neglected and various assumptions meant there was little incentive to isolate marine strains for search and discovery of new drugs...

  15. Diversity and bioprospecting of actinomycete endophytes from the medicinal plants.

    Science.gov (United States)

    Nalini, M S; Prakash, H S

    2017-04-01

    The endophytic actinomycetes constitute one of the fascinating group of microorganisms associated with a wide range of plant species. The diversity of actinomycetes in plants and their tissue parts is a matter of debate as no consensus are derived between individual studies. Nevertheless, their diversity correlates with the occurrence in plant species harboured in unique regions of biologically diverse areas called "hot spots." Recent advances in the isolation techniques have facilitated the isolation of rare taxa from these environments. The biosynthetic ability of the endophytic actinomycetes has proven beyond doubt that these organisms have the potential to synthesize an array of compounds with novelty in structure and bioactivity and as a result are preferred in the natural product screening programs. In the years to come, the scientific world may await to discover many more novel actinomycete taxa with metabolic diversity and applications in therapeutics.

  16. Application of actinomycetes to soil to ameliorate water repellency.

    Science.gov (United States)

    McKenna, F; El-Tarabily, K A; Petrie, S; Chen, C; Dell, B

    2002-01-01

    The aim of this study was to develop a novel isolation technique using a mixture of Bacillus and Streptomyces phages to selectively isolate wax-utilizing non-streptomycete actinomycetes effective in ameliorating water repellency in a problem soil. Phages added to a soil suspension reduced the dominance of Bacillus and Streptomyces isolates and significantly increased the number of non-streptomycete actinomycetes on isolation plates. Promising isolates, grown on a medium containing beeswax as sole carbon source, were selected for application to water repellent soil. Their addition significantly reduced water repellency. Phage application significantly increased the isolation of non-streptomycete actinomycetes. Wax-utilizing isolates were found to significantly reduce water repellency in a problem soil. The phage technique can be used for the routine isolation of non-streptomycete actinomycetes. Beeswax medium can be used to selectively isolate wax-utilizing micro-organisms with the potential to ameliorate water repellency in soil.

  17. Isolation and screening of actinomycetes from Sundarbans soil for ...

    African Journals Online (AJOL)

    USER

    2010-07-19

    Jul 19, 2010 ... Key words: Sundarbans, actinomycetes, isolate, antibacterial activity, identification. .... smears from colonies up to 10 days, stained by Gram's method as described ... Nutrient agar slants were used for short time preservation of.

  18. Extracellular overexpression of recombinant Thermobifida fusca cutinase by alpha-hemolysin secretion system in E. coli BL21(DE3

    Directory of Open Access Journals (Sweden)

    Su Lingqia

    2012-01-01

    Full Text Available Abstract Background Extracellular expression of proteins has an absolute advantage in a large-scale industrial production. In our previous study, Thermobifida fusca cutinase, an enzyme mainly utilized in textile industry, was expressed via type II secretory system in Escherichia coli BL21(DE3, and it was found that parts of the expressed protein was accumulated in the periplasmic space. Due to the fact that alpha-hemolysin secretion system can export target proteins directly from cytoplasm across both cell membrane of E. coli to the culture medium, thus in the present study we investigated the expression of cutinase using this alpha-hemolysin secretion system. Results T. fusca cutinase was fused with the specific signal peptide of alpha-hemolysin scretion system and expressed in E. coli BL21(DE3. In addition, HlyB and HlyD, strain-specific translocation components of alpha-hemolysin secretion system, were coexpressed to facilitate the enzyme expression. The cultivation of this engineered cell showed that cutinase activity in the culture medium reached 334 U/ml, which is 2.5 times that from type II secretion pathway under the same culture condition. The recombinant cutinase was further purified. Biochemical characterization of purified enzyme, which had an α-hemolysin secretion pathway signal peptide attached, had substrate specificity, pH and temperature profile, as well as application capability in bioscouring similar to that of wild-type cutinase. Conclusions In the present study, T. fusca cutinase was successfully secreted to the culture media by α-hemolysin secretion system. This is the first report of cutinase being efficiently secreted by this pathway. Due to the limited cases of successful expression of industrial enzyme by E. coli α-hemolysin secretion system, our study further explored the utilization of this pathway in industrial enzymes.

  19. Cellulolytic activity and structure of symbiotic bacteria in locust guts.

    Science.gov (United States)

    Su, L-J; Liu, H; Li, Y; Zhang, H-F; Chen, M; Gao, X-H; Wang, F-Q; Song, A-D

    2014-09-29

    Locusts are able to digest the cellulose of Gramineae plants, resulting in their being considered as major crop pests. To illustrate the mechanism involved in cellulose digestion, the cellulolytic activity and zymography in the gut contents of 16 locust species were determined using carboxymethyl cellulose (CMC) as substrate. The diversity of gut symbiotic bacteria was studied using denaturing gradient gel electrophoresis (DGGE). The results showed that high CMC activity was present in Acrididae gut fluid (mean 356.4 U/g proteins). Of the 5 locust species, Oxya chinensis had the highest diversity of intestinal symbiotic bacteria, characterized by the DGGE profile containing more than 20 bands of 16S rRNA. Klebsiella pneumoniae, in the gut of Locusta migratoria manilensis, was identified as the most abundant symbiotic bacterium by DNA sequencing, with a relative abundance of 19.74%. In comparison, Methylobacterium sp was the most dominant species in the Atractomorpha sinensis gut, with a relative abundance of 29.04%. The results indicated that the cellulolytic enzymes and gut microbial communities probably reflected their phylogenetic relationship with different locust species and associated feeding strategies.

  20. Cellulosomes - promising supramolecular machines of anaerobic cellulolytic microorganisms.

    Science.gov (United States)

    Vodovnik, Maša; Marinšek-Logar, Romana

    2010-12-01

    Cellulose is the main structural component of plant cell wall and thus the most abundant carbohydrate in nature. However, extracting the energy from this abundant source is limited by its recalcitrant nature. The hydrolysis of plant cell wall requires synergystic action of different enzymes, including multiple cellulases, hemicellulases, pectinases, etc. Meanwhile aerobic cellulolytic microorganisms release large quantities of synergistically acting free enzymes in their environment, most anaerobic microorganisms evolved more efficient strategies to optimize the process of plant cell wall degradation, for example production of extracellular multi-enzyme complexes (cellulosomes). Cellulosomes consist of at least one core structural protein, named scaffoldin, which firmly binds numerous enzymatic subunits, and usually also plays a major role in substrate binding. Although the general structure of cellulosomes seems universal, differences in number and identity of complex components do exist among microorganisms. The article surveys the current knowledge about cellulosomes, focusing on three best investigated cellulolytic clostridia, one representative of ruminal bacteria and novel findings concerning anaerobic fungi. Efforts in construction of artificially engineered cellulosomal systems (designer cellulosomes) as well as their biotechnological potential are also discussed.

  1. Production of cellulolytic enzymes by fungal cultures. [Aspergillus, Trichoderma, Chaetomium, Stachybotrys, and Hypocrea

    Energy Technology Data Exchange (ETDEWEB)

    Pyc, R.; Fiechter, A. Galas, E.

    1977-01-01

    Twelve fungal cultures belonging to the genera of Aspergillus, Trichoderma, Chaetomium, Stachybotrys, and Hypocrea were screened for the production of cellulolytic activity. All twelve were found to degrade xylan, avicel, and carboxymethylcellulose. More cellulolytic activity was obtained with shaken cultures than with still cultures and the addition of citrate-phosphate buffer to the media greatly depressed the levels of cellulolytic activity. Varying the composition of the mineral salts in the medium had no effect on the cellulolytic activity. The growth of Aspergillus wentii under controlled conditions in a bioreactor showed that the cellulolytic activity was not affected by the aeration rate or the type of stirrer. The rate of stirring, however, did effect the cellulolytic activity, as at lower stirring speeds considerable wall growth occurred which resulted in low levels of cellulolytic activity. Culture supernatant from Aspergillus wentii was found to hydrolyze from 30-32% of Solka-Floc and from 2-10% of corn cobs, wheat straw, and newsprint. The extensive hydrolysis of Solka-Floc indicates that with suitable treated cellulosic wastes and appropriate enzymes, appreciable amounts of sugars could be obtained.

  2. An efficient and improved methodology for the screening of industrially valuable xylano-pectino-cellulolytic microbes.

    Science.gov (United States)

    Singh, Avtar; Kaur, Amanjot; Dua, Anita; Mahajan, Ritu

    2015-01-01

    Xylano-pectino-cellulolytic enzymes are valuable enzymes of the industrial sector. In our earlier study, we have reported a novel and cost effective methodology for the qualitative screening of cellulase-free xylano-pectinolytic microorganisms by replacing the commercial, highly expensive substrates with agricultural residues, but the microorganisms with xylanolytic, pectinolytic, cellulolytic, xylano-pectinolytic, xylano-cellulolytic, pectino-cellulolytic, and xylano-pectino-cellulolytic potential were obtained. The probability of getting the desired combination was low, so efforts were made to further improve this cost effective methodology for obtaining the high yield of the microbes capable of producing desired combination of enzymes. By inclusion of multiple enrichment steps in sequence, using only practically low cost substrates and without any nutrient media till primary screening stage, this improved novel protocol for screening gave only the desired microorganisms with xylano-pectino-cellulolytic activity. Using this rapid, efficient, cost effective, and improved methodology, microbes with required combination of enzymes can be obtained and the probability of getting the desired microorganisms is cent percent. This is the first report presenting the methodology for the isolation of xylano-pectino-cellulolytic positive microorganisms at low cost and consuming less time.

  3. Compositions for enhancing hydroysis of cellulosic material by cellulolytic enzyme compositions

    Energy Technology Data Exchange (ETDEWEB)

    Quinlan, Jason; Xu, Feng; Sweeney, Matthew; Johansen, Katja Salomon

    2014-09-30

    The present invention relates to compositions comprising a GH61 polypeptide having cellulolytic enhancing activity and an organic compound comprising a carboxylic acid moiety, a lactone moiety, a phenolic moiety, a flavonoid moiety, or a combination thereof, wherein the combination of the GH61 polypeptide having cellulolytic enhancing activity and the organic compound enhances hydrolysis of a cellulosic material by a cellulolytic enzyme compared to the GH61 polypeptide alone or the organic compound alone. The present invention also relates to methods of using the compositions.

  4. Antimicrobial potential of Actinomycetes species isolated from marine environment.

    Science.gov (United States)

    Valli, S; Suvathi, Sugasini S; Aysha, O S; Nirmala, P; Vinoth, Kumar P; Reena, A

    2012-06-01

    To evaluate the antimicrobial activity of Actinomycetes species isolated from marine environment. Twenty one strains of Actinomycetes were isolated from samples of Royapuram, Muttukadu, Mahabalipuram sea shores and Adyar estuary. Preliminary screening was done using cross-streak method against two gram-positive and eight gram-negative bacteria. The most potent strains C11 and C12 were selected from which antibacterial substances were extracted. The antibacterial activities of the extracts were performed using Kirby-Bauer disc diffusion method. Molecular identification of those isolates was done. All those twenty one isolates were active against at least one of the test organisms. Morphological characters were recorded. C11 showed activity against Staphylococcus species (13.0±0.5 mm), Vibrio harveyi (11.0±0.2 mm), Pseudomonas species (12.0±0.3 mm). C12 showed activity against Staphylococcus species (16.0±0.4 mm), Bacillus subtilis (11.0±0.2 mm), Vibrio harveyi (9.0±0.1 mm), Pseudomonas species (10.0±0.2 mm). 16S rRNA pattern strongly suggested that C11 and C12 strains were Streptomyces species. The results of the present investigation reveal that the marine Actinomycetes from coastal environment are the potent source of novel antibiotics. Isolation, characterization and study of Actinomycetes can be useful in discovery of novel species of Actinomycetes.

  5. Bioprospecting potential of halogenases from Arctic marine actinomycetes.

    Science.gov (United States)

    Liao, Li; Chen, Ruiqin; Jiang, Ming; Tian, Xiaoqing; Liu, Huan; Yu, Yong; Fan, Chenqi; Chen, Bo

    2016-03-10

    Halometabolites, an important group of natural products, generally require halogenases for their biosynthesis. Actinomycetes from the Arctic Ocean have rarely been investigated for halogenases and their gene clusters associated, albeit great potential of halometabolite production has been predicted. Therefore, we initiated this research on the screening of halogenases from Arctic marine actinomycetes isolates to explore their genetic potential of halometabolite biosynthesis. Nine halogenase genes were discovered from sixty Arctic marine actinomycetes using in-house designed or previously reported PCR primers. Four representative genotypes were further cloned to obtain full coding regions through genome walking. The resulting halogenases were predicted to be involved in halogenation of indole groups, antitumor agent ansamitocin-like substrates, or unknown peptide-like compounds. Genome sequencing revealed a potential gene cluster containing the halogenase predicted to catalyze peptide-like compounds. However, the gene cluster was probably silent under the current conditions. PCR-based screening of halogenase genes is a powerful and efficient tool to conduct bioprospecting of halometabolite-producing actinomycetes from the Arctic. Genome sequencing can also identify cryptic gene clusters potentially producing new halometabolites, which might be easily missed by traditional isolation and chemical characterization. In addition, our study indicates that great genetic potential of new halometabolites can be expected from mostly untapped actinomycetes from the polar regions.

  6. Antimicrobial potential of Actinomycetes species isolated from marine environment

    Institute of Scientific and Technical Information of China (English)

    Valli S; Suvathi Sugasini S; Aysha OS; Nirmala P; Vinoth Kumar P; Reena A

    2012-01-01

    Objective:To evaluate the antimicrobial activity of Actinomycetes species isolated from marine environment. Methods: Twenty one strains of Actinomycetes were isolated from samples of Royapuram, Muttukadu, Mahabalipuram sea shores and Adyar estuary. Preliminary screening was done using cross-streak method against two gram-positive and eight gram-negative bacteria. The most potent strains C11 and C12 were selected from which antibacterial substances were extracted. The antibacterial activities of the extracts were performed using Kirby-Bauer disc diffusion method. Molecular identification of those isolates was done. Results:All those twenty one isolates were active against at least one of the test organisms. Morphological characters were recorded. C11 showed activity against Staphylococcus species (13.0±0.5 mm), Vibrio harveyi (11.0±0.2 mm), Pseudomonas species (12.0±0.3 mm). C12 showed activity against Staphylococcus species (16.0±0.4 mm), Bacillus subtilis (11.0±0.2 mm), Vibrio harveyi (9.0±0.1 mm), Pseudomonas species (10.0±0.2 mm). 16S rRNA pattern strongly suggested that C11 and C12 strains were Streptomyces species. Conclusions: The results of the present investigation reveal that the marine Actinomycetes from coastal environment are the potent source of novel antibiotics. Isolation, characterization and study of Actinomycetes can be useful in discovery of novel species of Actinomycetes.

  7. Antibacterial activity of some actinomycetes from Tamil Nadu, India

    Institute of Scientific and Technical Information of China (English)

    Pachaiyappan Saravana Kumar; John Poonga Preetam Raj; Veeramuthu Duraipandiyan; Savarimuthu Ignacimuthu

    2012-01-01

    Objective:To isolate novel actinomycetes and to evaluate their antibacterial activity. Methods:Three soil samples were collected from Vengodu (village) in Kanchipuram district, Tamil Nadu, India. Actinomycetes were isolated using serial dilution and plating method on actinomycetes isolation agar. Results: Totally 35 isolates were obtained on the basis of colony characteristics on actinomycetes isolation agar. All the isolates were screened for antibacterial activity by cross streak method. Medium and optimization of day were done for the potent strains using Nathan's agar well diffusion method. Isolation of bioactive compounds from significant active isolates was done by using different media. The most active isolate VAS 10 was identified as Actinobacterium Loyola PBT VAS 10 (accession No. JF501398) using 16s rRNA sequence method. The hexane, ethyl acetate, dichloromethane and butanol extracts of VAS 10 were tested against bacteria. The maximum antibacterial activity was observed in dichloromethane and ethyl acetate;maximum zones of inhibition were observed against Enterococcus durans. The rRNA secondary structure and the restriction sites of Actinobacterium Loyola VAS 10 were predicted using Genebee and NEBCutter online tools respectively. Conclusions: The present study showed that among the isolated actinomycetes, Actinobacterium Loyola PBT VAS 10 (accession No. JF501398) showed good antibacterial activity against the tested bacteria.

  8. SACCHARIFICATION OF CORNCOB USING CELLULOLYTIC BACTERIA FOR BIOETHANOL PRODUCTION

    Directory of Open Access Journals (Sweden)

    TITI CANDRA SUNARTI

    2010-08-01

    Full Text Available The use of cellulose degrading enzyme (cellulases for hydrolysis of lignocellulosic material is a part of bioethanol production process. In this experiment, delignified corncob, its cellulose fraction and alpha cellulose were used as substrates to produce fermentable sugar by using three local isolates of celluloytic bacteria (C5-1, C4-4, C11-1 and Cmix ; mixed cultures of three isolates, and Saccharomyces cereviseae to produce ethanol. The results showed that all isolates of cellulolytic bacteria can grow on cellulose fraction better than on delignified corncob, and alpha cellulose. The highest hydrolytic activity produced from cellulose fraction was by isolate C4-4, which liberated 3.50 g/l of total sugar. Ethanol can be produced by mixed culture of bacteria and yeast, but because of competitive growth, the fermentation only produced 0.39-0.47 g/l of ethanol.

  9. NREL Explains the Higher Cellulolytic Activity of a Vital Microorganism

    Energy Technology Data Exchange (ETDEWEB)

    2016-06-01

    The discovery of a new mode of action by C. thermocellum to convert biomass to biofuels is significant because the bacterium is already recognized as one of the most effective in the biosphere. Researchers found that, in addition to using common cellulase degradation mechanisms attached to cells, C. thermocellum also uses a new category of cell-free scaffolded enzymes. The new discovery will influence the strategies used to improve the cellulolytic activity of biomass degrading microbes going forward. Better understanding of this bacterium could lead to cheaper production of ethanol and drop-in fuels. Also, this discovery demonstrates that nature's biomass conversion behaviors are not fully understood and remain as opportunities for future microbial/enzyme engineering efforts.

  10. Synthetic polyester-hydrolyzing enzymes from thermophilic actinomycetes.

    Science.gov (United States)

    Wei, Ren; Oeser, Thorsten; Zimmermann, Wolfgang

    2014-01-01

    Thermophilic actinomycetes produce enzymes capable of hydrolyzing synthetic polyesters such as polyethylene terephthalate (PET). In addition to carboxylesterases, which have hydrolytic activity predominantly against PET oligomers, esterases related to cutinases also hydrolyze synthetic polymers. The production of these enzymes by actinomycetes as well as their recombinant expression in heterologous hosts is described and their catalytic activity against polyester substrates is compared. Assays to analyze the enzymatic hydrolysis of synthetic polyesters are evaluated, and a kinetic model describing the enzymatic heterogeneous hydrolysis process is discussed. Structure-function and structure-stability relationships of actinomycete polyester hydrolases are compared based on molecular dynamics simulations and recently solved protein structures. In addition, recent progress in enhancing their activity and thermal stability by random or site-directed mutagenesis is presented.

  11. Hybrid isoprenoid secondary metabolite production in terrestrial and marine actinomycetes.

    Science.gov (United States)

    Gallagher, Kelley A; Fenical, William; Jensen, Paul R

    2010-12-01

    Terpenoids are among the most ubiquitous and diverse secondary metabolites observed in nature. Although actinomycete bacteria are one of the primary sources of microbially derived secondary metabolites, they rarely produce compounds in this biosynthetic class. The terpenoid secondary metabolites that have been discovered from actinomycetes are often in the form of biosynthetic hybrids called hybrid isoprenoids (HIs). HIs include significant structural diversity and biological activity and thus are important targets for natural product discovery. Recent screening of marine actinomycetes has led to the discovery of a new lineage that is enriched in the production of biologically active HI secondary metabolites. These strains represent a promising resource for natural product discovery and provide unique opportunities to study the evolutionary history and ecological functions of an unusual group of secondary metabolites. Copyright © 2010 Elsevier Ltd. All rights reserved.

  12. CRISPR-Cas9 Based Engineering of Actinomycetal Genomes

    DEFF Research Database (Denmark)

    Tong, Yaojun; Charusanti, Pep; Zhang, Lixin

    2015-01-01

    . To facilitate the genetic manipulation of actinomycetes, we developed a highly efficient CRISPR-Cas9 system to delete gene(s) or gene cluster(s), implement precise gene replacements, and reversibly control gene expression in actinomycetes. We demonstrate our system by targeting two genes, actIORF1 (SCO5087......) and actVB (SCO5092), from the actinorhodin biosynthetic gene cluster in Streptomyces coelicolor A3(2). Our CRISPR-Cas9 system successfully inactivated the targeted genes. When no templates for homology-directed repair (HDR) were present, the site-specific DNA double-strand breaks (DSBs) introduced by Cas9....... Moreover, we developed a system to efficiently and reversibly control expression of target genes, deemed CRISPRi, based on a catalytically dead variant of Cas9 (dCas9). The CRISPR-Cas9 based system described here comprises a powerful and broadly applicable set of tools to manipulate actinomycetal genomes....

  13. Alkalithermophilic actinomycetes in a subtropical area of Jujuy, Argentina.

    Science.gov (United States)

    Carrillo, L; Benítez Ahrendts, M R; Maldonado, M J

    2009-01-01

    The objective of this study was to examine the alkalithermophilic actinomycete communities in the subtropical environment of Jujuy, Argentina, characterized by sugarcane crops. Laceyella putida, Laceyella sacchari, Thermoactinomyces intermedius, Thermoactinomyces vulgaris and Thermoflavimicrobium dichotomicum were isolated on the media with novobiocin, from sugar cane plants and renewal rhizospheres, and grass and wood soils. Soil pH was almost neutral or lightly alkaline, except for grass soil acidified by lactic liquor. A smaller number of actinomycetes was found on the living plants and bagasse (recently obtained or stored according to the Ritter method) with respect to decomposed leaves on the soil. Thermophilic species of Laceyella, Thermoactinomyces, Thermoflavimicrobium, Saccharomonospora, Streptomyces and Thermononospora were isolated on the media without novobiocin, from composted sugar cane residues. Air captured near composted bagasse piles, contained alkalithermophilic actinomycete spores.

  14. The significance of cellulolytic enzymes produced by Trichoderma in opportunistic lifestyle of this fungus.

    Science.gov (United States)

    Strakowska, Judyta; Błaszczyk, Lidia; Chełkowski, Jerzy

    2014-07-01

    The degradation of native cellulose to glucose monomers is a complex process, which requires the synergistic action of the extracellular enzymes produced by cellulolytic microorganisms. Among fungi, the enzymatic systems that can degrade native cellulose have been extensively studied for species belonging to the genera of Trichoderma. The majority of the cellulolytic enzymes described so far have been examples of Trichoderma reesei, extremely specialized in the efficient degradation of plant cell wall cellulose. Other Trichoderma species, such as T. harzianum, T. koningii, T. longibrachiatum, and T. viride, known for their capacity to produce cellulolytic enzymes, have been isolated from various ecological niches, where they have proved successful in various heterotrophic interactions. As saprotrophs, these species are considered to make a contribution to the degradation of lignocellulosic plant material. Their cellulolytic potential is also used in interactions with plants, especially in plant root colonization. However, the role of cellulolytic enzymes in species forming endophytic associations with plants or in those existing in the substratum for mushroom cultivation remains unknown. The present review discusses the current state of knowledge about cellulolytic enzymes production by Trichoderma species and the encoding genes, as well as the involvement of these proteins in the lifestyle of Trichoderma.

  15. Biodegradation of the herbicide Diuron in soil by indigenous actinomycetes.

    Science.gov (United States)

    Esposito, E; Paulillo, S M; Manfio, G P

    1998-08-01

    Three actinomycete strains isolated from soil treated with 2,4-D were able to degrade the herbicide Diuron in vitro. Strain CCT 4916 was the most efficient, degrading up to 37% of applied Diuron (100 mg Kg-1 soil) in 7 days, as measured by HPLC and UV/VIS spectroscopy. All strains showed protease and urease activity; intracellular activity of metapyrocatechase and pyrocatechase were not found. Actinomycete strain CCT 4916 produced manganese peroxidase, which could be potentially related to degradation of Diuron.

  16. Antibiotic production by actinomycetes: the Janus faces of regulation.

    Science.gov (United States)

    Cundliffe, Eric

    2006-07-01

    This manuscript reviews some of the common regulatory mechanisms that control antibiotic production in actinomycetes. These ubiquitous bacteria, collectively responsible for the earthy smell of soil, are prolific producers of antibiotics and other secondary metabolites. The content of this review is biased towards the author's current research interests, concerning the action of regulatory gene products that control transcription of antibiotic-biosynthetic genes and the associated involvement of low molecular weight signalling molecules of the gamma-butyrolactone family. As a result, much fertile ground remains unturned particularly in the area of environmental monitoring and responses of actinomycetes to stimuli so perceived. Reviews casting a broader net are cited in the text.

  17. ISOLATION OF ENDOPHYTIC ACTINOMYCETES FROM MEDICINAL PLANTS AND ITS MUTATIONAL EFFECT IN BIOCONTROL ACTIVITY

    Directory of Open Access Journals (Sweden)

    Hema Shenpagam N.*, D. Kanchana Devi ** and Sinduja G.

    2012-11-01

    Full Text Available In the present study, the endophytic actinomycetes were collected from three medicinal plants Azadiracta indica, Ocimum sanctum and Phyllanthus amarus. Endophytic actinomycetes were isolated using different media like Starch casein agar, Starch casein nitrate agar, Actinomycetes isolation agar and Soyabean agar, while it showed more colonies in Starch casein agar. The endophytic actinomycetes were stained and biochemical tests were performed. Antimicrobial compound was purified from the filtrate by ethanol extraction method. Antagonistic activities of endophytic actinomycetes isolates were tested against bacterial pathogens (Staphylococcus aureus, Streptococcus pyogenes, Klebsiella pneumoniae and Pseudomonas aeruginosa and the fungi Rhizopus. For the selected isolates antibiotic resistance was checked using various antibiotic discs like Amoxycillin, Penicillin, Rifampicin and Ampicillin. The strains which showed efficient antibacterial activity were selected to study the effect of mutation by physical and chemical method. In this study, UV mutated endophytic actinomycetes increase antibiotic production than non-mutated endophytic Actinomycetes, whereas in chemical mutation it does not increase the antibiotic production.

  18. Biodiscovery from rare actinomycetes: an eco-taxonomical perspective.

    Science.gov (United States)

    Kurtböke, D I

    2012-03-01

    Microbial natural products, in particular, the ones produced by the members of the order Actinomycetales, will continue to represent an important route to the discovery of novel classes of bioactive compounds. As a result, the search for and discovery of lesser-known and/or novel actinomycetes is of significant interest to the industry due to a growing need for the development of new and potent therapeutic agents, mainly against drug resistant bacteria. Current advancements in genomics and metagenomics are adding strength to the target-directed search for detection and isolation of bioactive actinomycetes. New discoveries, however, will only stem from a sound understanding and interpretation of knowledge derived from conventional studies conducted since the discovery of streptomycin, on the ecology, taxonomy, physiology and metabolism of actinomycetes, and from a combination of this knowledge with currently available and continuously advancing molecular tools. Such a powerful information platform will then inevitably reveal the whereabouts, taxonomical and chemical identities of previously undetected bioactive actinomycetes including novel species of streptomycetes as potential producers of novel drug candidates.

  19. Identification and Antimicrobial Properties of Malaysian Mangrove Actinomycetes

    Directory of Open Access Journals (Sweden)

    Suhaidi Ariffin

    2017-02-01

    Full Text Available The mangrove area is one of the more unique and less studied habitats for the discovery of biologically active secondary metabolites from potential actinomycetes. For this reason, isolation, characterization and screening of antimicrobial properties of actinomycetes from mangrove soils collected at the selected mangrove areas in Malaysia were conducted. A total of 73 actinomycetes were isolated. Of these, 9.6% (direct broth culture and 8.2% (ethyl acetate extract of these isolates demonstrated antimicrobial activities active against Staphylococcus aureus, Bacillus subtilis, Candida albicans as well as Saccharomyces cerevisiae by disc diffusion assay. Hexane extracts, however, exhibited no activity. A modified microdilution plate assay showed that 16.4% (ethyl acetate extract of these strains were able to generate antimicrobial compounds active against Staphylococcus aureus, Bacillus subtilis, and Candida albicans. In addition, morphological observations and 16S rRNA sequence analysis indicate the presence of representative species from at least 3 genera Streptomyces, Pseudonocardia and Saccharomonospora. Thus, Malaysian mangrove soils could be an interesting source to explore for antimicrobial secondary metabolites and considered to have a diversity of marine actinomycetes.

  20. The function of vesicles in the actinomycete Frankia

    NARCIS (Netherlands)

    Meesters, T.

    1988-01-01

    The actinomycete Frankia is a symbiotic nitrogen fixer, living in root nodules of many non-leguminous plants. A typical characteristic of this endophytic organism is the formation of specialized swollen cell structures, called vesicles. Frankia

  1. Antibiotics production by an actinomycete isolated from the termite gut.

    Science.gov (United States)

    Matsui, Toru; Tanaka, Junichi; Namihira, Tomoyuki; Shinzato, Naoya

    2012-12-01

    As well as the search for new antibiotics, a new resource or strains for the known antibiotics is also important. Microbial symbionts in the gut of termites could be regarded as one of the feasible resource for such purpose. In this study, antibiotic-producing actinomycetes were screened from symbionts of the termite gut. 16SrRNA sequence analysis for the 10 isolates revealed that they belong to actinomycetes such as Streptomyces sp., Kitasatospora sp., and Mycobacterium sp. A culture broth from one of the isolate, namely strain CA1, belonging to the genera Streptomyces exhibited antagonistic activity against actinomycetes (Micrococcus spp.), gram-positive bacteria (Bacillus spp.), and yeast (Candida spp.). The structures of 2 compounds isolated from the culture broth of the strain CA1 were identified as those of actinomycin X2 and its analog, D. This study is the first to report that some symbionts of the termite gut are antibiotic-producing actinomycetes, and suggest that the termite gut is a feasible resource for bioprospecting.

  2. The function of vesicles in the actinomycete Frankia

    NARCIS (Netherlands)

    Meesters, T.

    1988-01-01

    The actinomycete Frankia is a symbiotic nitrogen fixer, living in root nodules of many non-leguminous plants. A typical characteristic of this endophytic organism is the formation of specialized swollen cell structures, called vesicles. Frankia vesi

  3. A New Degraded Sesquiterpene from Marine Actinomycete Streptomyces sp. 0616208

    Institute of Scientific and Technical Information of China (English)

    Xiu Chao XIE; Wen Li MEI; You Xing ZHAO; Kui HONG; Hao Fu DAI

    2006-01-01

    A new degraded sesquiterpene was isolated from the marine actinomycete Streptomyces sp. 0616208. Its structure was elucidated as (1α, 4aα, 5α, 7β, 8aβ)-5, 8a-dimethyl-decahydrona-phthalene-1, 4a, 7-triol on the basis of spectroscopic data.

  4. Environmental and metabolomic study of antibiotic production by actinomycetes

    NARCIS (Netherlands)

    Zhu, Hua

    2014-01-01

    This thesis may be regarded as a concept work, to see how feasible drug discovery approaches still are. For this, a strain collection was built up consisting of actinomycetes from soil in the Qinling and Himalaya mountains, which were subsequently tested for antibiotic production against multi-drug

  5. Environmental and metabolomic study of antibiotic production by actinomycetes

    NARCIS (Netherlands)

    Zhu, Hua

    2014-01-01

    This thesis may be regarded as a concept work, to see how feasible drug discovery approaches still are. For this, a strain collection was built up consisting of actinomycetes from soil in the Qinling and Himalaya mountains, which were subsequently tested for antibiotic production against multi-drug

  6. Isolasi Actinomycetes Dengan Menggunakan Metode Skrining Sebagai Penghasil Enzim Kitinase

    Directory of Open Access Journals (Sweden)

    Welly Anggraini

    2015-04-01

    Full Text Available Penelitian ini bertujuan untuk mengisolasi Actinomycetes yang memiliki kemampuan dalam mendegradasi kitin.  Isolasi Actinomycetes dilakukan menggunakan media NaST21Cx selama 2-3 bulan dan purifikasi untuk mendapatkan isolat murni dilakukan menggunakan media ISP-2. Hasil dari penelitian ini yakni, empat isolat Actinomycetes, yang diberi kode ANL-4, ANL-9, ANL-12, dan ANL-d-2b-3 yang ditandai dengan ciri-ciri isolat yang keras dan melekat erat pada agar.  Secara mikroskopis teramati adanya hifa dan spora yang terbentuk.  Isolat ANL-4, ANL-9, ANL-12, dan ANLd-2b-3 memiliki kemampuan dalam mendegradasi kitin pada media uji mineral-salt agar plate dengan substrat kitin 1% (w/v yang diindikasikan dengan terbentukya zona bening yang diperjelas dengan penambahan Congo Red 1% (w/v.  Indeks kitinolitik yang dihasilkan berturut-turut : 5 cm, 2 cm, 1,9 cm, dan 2,3 cm. Kata Kunci : actinomycetes, hutan bakau, kitin, kitinase

  7. Insight into Dominant Cellulolytic Bacteria from Two Biogas Digesters and Their Glycoside Hydrolase Genes.

    Directory of Open Access Journals (Sweden)

    Yongjun Wei

    Full Text Available Diverse cellulolytic bacteria are essential for maintaining high lignocellulose degradation ability in biogas digesters. However, little was known about functional genes and gene clusters of dominant cellulolytic bacteria in biogas digesters. This is the foundation to understand lignocellulose degradation mechanisms of biogas digesters and apply these gene resource for optimizing biofuel production. A combination of metagenomic and 16S rRNA gene clone library methods was used to investigate the dominant cellulolytic bacteria and their glycoside hydrolase (GH genes in two biogas digesters. The 16S rRNA gene analysis revealed that the dominant cellulolytic bacteria were strains closely related to Clostridium straminisolvens and an uncultured cellulolytic bacterium designated BG-1. To recover GH genes from cellulolytic bacteria in general, and BG-1 in particular, a refined assembly approach developed in this study was used to assemble GH genes from metagenomic reads; 163 GH-containing contigs ≥ 1 kb in length were obtained. Six recovered GH5 genes that were expressed in E. coli demonstrated multiple lignocellulase activities and one had high mannanase activity (1255 U/mg. Eleven fosmid clones harboring the recovered GH-containing contigs were sequenced and assembled into 10 fosmid contigs. The composition of GH genes in the 163 assembled metagenomic contigs and 10 fosmid contigs indicated that diverse GHs and lignocellulose degradation mechanisms were present in the biogas digesters. In particular, a small portion of BG-1 genome information was recovered by PhyloPythiaS analysis. The lignocellulase gene clusters in BG-1 suggested that it might use a possible novel lignocellulose degradation mechanism to efficiently degrade lignocellulose. Dominant cellulolytic bacteria of biogas digester possess diverse GH genes, not only in sequences but also in their functions, which may be applied for production of biofuel in the future.

  8. Insight into Dominant Cellulolytic Bacteria from Two Biogas Digesters and Their Glycoside Hydrolase Genes.

    Science.gov (United States)

    Wei, Yongjun; Zhou, Haokui; Zhang, Jun; Zhang, Lei; Geng, Alei; Liu, Fanghua; Zhao, Guoping; Wang, Shengyue; Zhou, Zhihua; Yan, Xing

    2015-01-01

    Diverse cellulolytic bacteria are essential for maintaining high lignocellulose degradation ability in biogas digesters. However, little was known about functional genes and gene clusters of dominant cellulolytic bacteria in biogas digesters. This is the foundation to understand lignocellulose degradation mechanisms of biogas digesters and apply these gene resource for optimizing biofuel production. A combination of metagenomic and 16S rRNA gene clone library methods was used to investigate the dominant cellulolytic bacteria and their glycoside hydrolase (GH) genes in two biogas digesters. The 16S rRNA gene analysis revealed that the dominant cellulolytic bacteria were strains closely related to Clostridium straminisolvens and an uncultured cellulolytic bacterium designated BG-1. To recover GH genes from cellulolytic bacteria in general, and BG-1 in particular, a refined assembly approach developed in this study was used to assemble GH genes from metagenomic reads; 163 GH-containing contigs ≥ 1 kb in length were obtained. Six recovered GH5 genes that were expressed in E. coli demonstrated multiple lignocellulase activities and one had high mannanase activity (1255 U/mg). Eleven fosmid clones harboring the recovered GH-containing contigs were sequenced and assembled into 10 fosmid contigs. The composition of GH genes in the 163 assembled metagenomic contigs and 10 fosmid contigs indicated that diverse GHs and lignocellulose degradation mechanisms were present in the biogas digesters. In particular, a small portion of BG-1 genome information was recovered by PhyloPythiaS analysis. The lignocellulase gene clusters in BG-1 suggested that it might use a possible novel lignocellulose degradation mechanism to efficiently degrade lignocellulose. Dominant cellulolytic bacteria of biogas digester possess diverse GH genes, not only in sequences but also in their functions, which may be applied for production of biofuel in the future.

  9. Insight into Dominant Cellulolytic Bacteria from Two Biogas Digesters and Their Glycoside Hydrolase Genes

    Science.gov (United States)

    Zhang, Jun; Zhang, Lei; Geng, Alei; Liu, Fanghua; Zhao, Guoping; Wang, Shengyue; Zhou, Zhihua; Yan, Xing

    2015-01-01

    Diverse cellulolytic bacteria are essential for maintaining high lignocellulose degradation ability in biogas digesters. However, little was known about functional genes and gene clusters of dominant cellulolytic bacteria in biogas digesters. This is the foundation to understand lignocellulose degradation mechanisms of biogas digesters and apply these gene resource for optimizing biofuel production. A combination of metagenomic and 16S rRNA gene clone library methods was used to investigate the dominant cellulolytic bacteria and their glycoside hydrolase (GH) genes in two biogas digesters. The 16S rRNA gene analysis revealed that the dominant cellulolytic bacteria were strains closely related to Clostridium straminisolvens and an uncultured cellulolytic bacterium designated BG-1. To recover GH genes from cellulolytic bacteria in general, and BG-1 in particular, a refined assembly approach developed in this study was used to assemble GH genes from metagenomic reads; 163 GH-containing contigs ≥ 1 kb in length were obtained. Six recovered GH5 genes that were expressed in E. coli demonstrated multiple lignocellulase activities and one had high mannanase activity (1255 U/mg). Eleven fosmid clones harboring the recovered GH-containing contigs were sequenced and assembled into 10 fosmid contigs. The composition of GH genes in the 163 assembled metagenomic contigs and 10 fosmid contigs indicated that diverse GHs and lignocellulose degradation mechanisms were present in the biogas digesters. In particular, a small portion of BG-1 genome information was recovered by PhyloPythiaS analysis. The lignocellulase gene clusters in BG-1 suggested that it might use a possible novel lignocellulose degradation mechanism to efficiently degrade lignocellulose. Dominant cellulolytic bacteria of biogas digester possess diverse GH genes, not only in sequences but also in their functions, which may be applied for production of biofuel in the future. PMID:26070087

  10. Characterization of cellulolytic activity from digestive fluids of Dissosteira carolina (Orthoptera: Acrididae).

    Science.gov (United States)

    Willis, Jonathan D; Klingeman, William E; Oppert, Cris; Oppert, Brenda; Jurat-Fuentes, Juan L

    2010-11-01

    Previous screening of head-derived and gut fluid extracts of Carolina grasshoppers, Dissosteira carolina (L.) revealed relatively high activity against cellulase substrates when compared to other insect groups. In this work we report on the characterization and identification of enzymes involved in cellulolytic activity in digestive fluids of D. carolina. In zymograms using carboxymethylcellulose (CMC) as substrate, we detected four distinct cellulolytic protein bands in D. carolina gut fluids, common to all developmental stages. These cellulolytic enzymes were localized to foregut and midgut regions of the D. carolina digestive tract. Cellulases were purified from D. carolina head and gut fluid extracts by liquid chromatography to obtain N-terminal amino acid sequence tags. Database searches with sequence tags from head fluids indicated high similarity with invertebrate, bacterial and plant beta1,4-endoglucanases, while no homologues were identified for the gut-derived protein. Our data demonstrate the presence of cellulolytic activity in the digestive system of D. carolina and suggest that cellulases of endogenous origin are present in this organism. Considering that this grasshopper species is a pest of grasses, including switchgrass that has been suggested bioethanol feedstock, characterization of insect cellulolytic systems may aid in developing applications for plant biomass biodegradation for biofuel production.

  11. Cellulolytic Bacteria Associated with the Gut of Dendroctonus armandi Larvae (Coleoptera: Curculionidae: Scolytinae

    Directory of Open Access Journals (Sweden)

    Xia Hu

    2014-03-01

    Full Text Available The object of this study was to investigate the cellulolytic bacterial community in the intestine of the Chinese white pine beetle (Dendroctonus armandi larvae. A total of 91 cellulolytic bacteria were isolated and assigned to 11 genotypes using amplified ribosomal DNA restriction analysis (ARDRA. Partial 16S rDNA sequence analysis and morphological tests were used to assign the 11 representative isolates. The results showed that the isolates belonged to α-Proteobacteria, γ-Proteobacteria and Firmicutes. Members of γ-Proteobacteria were the most frequently represented species and accounted for 73.6% of all the cellulolytic bacteria. The majority of cellulolytic bacteria in D. armandi larva gut were identified as Serratia and accounted for 49.5%, followed by Pseudomonas, which accounted for 22%. In addition, members of Bacillus, Brevundimonas, Paenibacillus, Pseudoxanthomonas, Methylobacterium and Sphingomonas were found in the D. armandi larva gut. Brevundimonas kwangchunensis, Brevundimonas vesicularis, Methylobacterium populi and Pseudoxanthomonas mexicana were reported to be cellulolytic for the first time in this study. Information generated from the present study might contribute towards understanding the relationship between bark beetle and its gut flora.

  12. Isolation and Identification of Cellulolytic Bacteria from the Gut of Three Phytophagus Insect Species

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    Rajib Kumar Shil

    2014-12-01

    Full Text Available The cellulolytic bacteria from the gut of three different phytophagous insects were studied to isolate novel cellulolytic organism for biofuel industry. Among the threse, gut of P. quatuordecimpunctata larvae contained both highest no of total bacterial count (6.8x107CFU/gut and cellulolytic bacteria (5.42x103CFU/gut. Fifteen different isolates were obtained from the gut of O. velox, A. miliarisand P. quatuordecimpunctata. All the isolates produced clear zone in CMC medium staining with Congo red. The isolates included Gram positive Enterococcus, Microbacterium and Gram negative Aeromonas, Erwinia, Serretia, Flavobacterium, Acenitobacter, Klebsiella, Yersinia, Xenorhabdus, Psedomonas and Photorhabdus. Out of the fifteen isolated and identified bacterial species, twelve bacterial species were novel being reported for first time as having cellulase activity.

  13. Parameter estimation for models of ligninolytic and cellulolytic enzyme kinetics

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Gangsheng [ORNL; Post, Wilfred M [ORNL; Mayes, Melanie [ORNL; Frerichs, Joshua T [ORNL; Jagadamma, Sindhu [ORNL

    2012-01-01

    While soil enzymes have been explicitly included in the soil organic carbon (SOC) decomposition models, there is a serious lack of suitable data for model parameterization. This study provides well-documented enzymatic parameters for application in enzyme-driven SOC decomposition models from a compilation and analysis of published measurements. In particular, we developed appropriate kinetic parameters for five typical ligninolytic and cellulolytic enzymes ( -glucosidase, cellobiohydrolase, endo-glucanase, peroxidase, and phenol oxidase). The kinetic parameters included the maximum specific enzyme activity (Vmax) and half-saturation constant (Km) in the Michaelis-Menten equation. The activation energy (Ea) and the pH optimum and sensitivity (pHopt and pHsen) were also analyzed. pHsen was estimated by fitting an exponential-quadratic function. The Vmax values, often presented in different units under various measurement conditions, were converted into the same units at a reference temperature (20 C) and pHopt. Major conclusions are: (i) Both Vmax and Km were log-normal distributed, with no significant difference in Vmax exhibited between enzymes originating from bacteria or fungi. (ii) No significant difference in Vmax was found between cellulases and ligninases; however, there was significant difference in Km between them. (iii) Ligninases had higher Ea values and lower pHopt than cellulases; average ratio of pHsen to pHopt ranged 0.3 0.4 for the five enzymes, which means that an increase or decrease of 1.1 1.7 pH units from pHopt would reduce Vmax by 50%. (iv) Our analysis indicated that the Vmax values from lab measurements with purified enzymes were 1 2 orders of magnitude higher than those for use in SOC decomposition models under field conditions.

  14. PGASO: A synthetic biology tool for engineering a cellulolytic yeast

    Directory of Open Access Journals (Sweden)

    Chang Jui-Jen

    2012-07-01

    Full Text Available Abstract Background To achieve an economical cellulosic ethanol production, a host that can do both cellulosic saccharification and ethanol fermentation is desirable. However, to engineer a non-cellulolytic yeast to be such a host requires synthetic biology techniques to transform multiple enzyme genes into its genome. Results A technique, named Promoter-based Gene Assembly and Simultaneous Overexpression (PGASO, that employs overlapping oligonucleotides for recombinatorial assembly of gene cassettes with individual promoters, was developed. PGASO was applied to engineer Kluyveromycesmarxianus KY3, which is a thermo- and toxin-tolerant yeast. We obtained a recombinant strain, called KR5, that is capable of simultaneously expressing exoglucanase and endoglucanase (both of Trichodermareesei, a beta-glucosidase (from a cow rumen fungus, a neomycin phosphotransferase, and a green fluorescent protein. High transformation efficiency and accuracy were achieved as ~63% of the transformants was confirmed to be correct. KR5 can utilize beta-glycan, cellobiose or CMC as the sole carbon source for growth and can directly convert cellobiose and beta-glycan to ethanol. Conclusions This study provides the first example of multi-gene assembly in a single step in a yeast species other than Saccharomyces cerevisiae. We successfully engineered a yeast host with a five-gene cassette assembly and the new host is capable of co-expressing three types of cellulase genes. Our study shows that PGASO is an efficient tool for simultaneous expression of multiple enzymes in the kefir yeast KY3 and that KY3 can serve as a host for developing synthetic biology tools.

  15. Technique for preparation of anaerobic microbes: Rodshaped cellulolytic bacteria

    Directory of Open Access Journals (Sweden)

    Amlius Thalib

    2001-10-01

    Full Text Available Preparation of anaerobic-rod cellulolytic bacteria with coating technique has been conducted. Steps of the processes involved were cultivation, coating, evaporation, and drying. Coating agent used was Gum Arabic, and drying techniquesconducted were freeze drying and sun drying. pH of culture media was firstly optimized to obtain the maximal population ofbacteria. Both coated and uncoated preparates were subjected to drying. Morphological and Gram type identifications showed that uncoated preparate dried with freeze drying is not contaminated (ie. all bacteria are rod shape with Gram-negative type while the one dried with sun drying is not morphologically pure (ie. containing of both rod and coccus shapes with Gram negative and positive. The coated preparates dried by both freeze and sun drying, were not contaminated (ie. all are rods with Gram-negative. The coating and drying processes decreased viability of preparates significantly. However, the decreasing of viability of coated preparate are lower than uncoated preparate (ie. 89 vs. 97%. Total count of bacteria in sun-drying coated preparate are higher (P<0.05 than the uncoated preparate (ie. 3.38 x 1010 vs. 1.97 x 1010 colony/g DM. Activity of sun-drying coated preparate to digest elephant grass and rice straw was higher (P<0.01 than the sun-drying uncoated preparate with the in vitro DMD values were 42.7 vs. 35.5% for elephant grass substrate and 29.3 vs. 24.6% for rice straw substrate. Therefore, it is concluded that coating technique has a positive effects on the preparation of rumen bacteria.

  16. Distribution of actinomycetes in oil contaminated ultisols of the Niger Delta (Nigeria)

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    The distribution of actinomycetes in oil contaminated sandy loam ultisols of the Niger Delta region of Nigeria was studied to aid in understanding the effect of hydrocarbons on indigenous microbial population in tropical soils. The average total counts of actinomycetes in all the oil samples analysed was 103 cfu/g. Higher counts of actinomycetes were observed during the dry season than during the wet season. The counts of hydrocarbonoclastic actinomycetes correlated positively with the total count of actinomycetes.The actinomycetes were generally restricted to the top soil(0-10 cm soil depth) although a seemingly deeper(down to 40 cm soil depth) distribution was noticed in the dry season. The isolates included oil degrading species of Actinoplanes, Norcadia,Streptomyces and Streptosporangium. Their high oil utilization ability indicates their positive potential and role in the bioremediation of oil-spilled soils.

  17. Isolation and characterization of cellulolytic bacteria from the Stain house Lake, Antarctica.

    Science.gov (United States)

    Melo, Itamar S; Zucchi, Tiago D; Silva, Rafael E; Vilela, Elke S D; Sáber, Mirian Lobo; Rosa, Luiz H; Pellizari, Vivian H

    2014-07-01

    The main aim was to evaluate the occurrence of cellulolytic bacteria from the Stain house Lake, located at Admiralty Bay, Antarctica. Thick cotton string served as a cellulose bait for the isolation of bacteria. A total of 52 bacterial isolates were recovered and tested for their cellulase activity, and two of them, isolates CMAA 1184 and CMAA 1185, showed significant cellulolytic activity on carboxymethylcellulose agar plates. Phylogenetic analysis placed the isolates into the Bacillus 16S ribosomal RNA gene subclade. Both isolates produced a cold-active cellulase which may play a crucial role in this extreme environment.

  18. Diversity, abundance and natural products of marine sponge-associated actinomycetes.

    Science.gov (United States)

    Abdelmohsen, Usama Ramadan; Bayer, Kristina; Hentschel, Ute

    2014-03-01

    Actinomycetes are known for their unprecedented ability to produce novel lead compounds of clinical and pharmaceutical importance. This review focuses on the diversity, abundance and methodological approaches targeting marine sponge-associated actinomycetes. Additionally, novel qPCR data on actinomycete abundances in different sponge species and other environmental sources are presented. The natural products literature is covered, and we are here reporting on their chemical structures, their biological activities, as well as the source organisms from which they were isolated.

  19. Unusual multifocal granulomatous disease caused by actinomycetous bacteria in a nestling Derbyan parrot (Psittacula derbiana).

    Science.gov (United States)

    Park, F J; Jaensch, S

    2009-01-01

    A nestling Derbyan parrot (Psittacula derbiana) was presented with unusual subcutaneous swellings of the thigh regions, and poor growth. Histological examination revealed actinomycetous bacteria associated with multifocal systemic granulomas. The clinical and pathological findings of the case are presented, and some relevant aspects of actinomycetous bacterial infections in mammals and birds are discussed. Although granulomatous disease is encountered at times in avian species, the actinomycetous bacteria (Nocardia and Actinomyces spp.) have rarely been reported in association with multifocal granulomatous disease in birds.

  20. 放线菌分类研究进展%Advance on taxonomy of actinomycetes

    Institute of Scientific and Technical Information of China (English)

    刘燕娟; 李婵; 周倩

    2009-01-01

    The status of actinomycetes classification study of the evolution and classification methods were overviewed, as well as domestic actinomycetes classification status,and actinomycetes classification was forecasted.%综述了放线菌分类地位的演变和分类研究方法,以及国内放线菌分类现状,并对放线菌分类研究进行了展望.

  1. Production of Ramoplanin and Ramoplanin Analogs by Actinomycetes

    Science.gov (United States)

    de la Cruz, Mercedes; González, Ignacio; Parish, Craig A.; Onishi, Russell; Tormo, José R.; Martín, Jesús; Peláez, Fernando; Zink, Debbie; El Aouad, Noureddine; Reyes, Fernando; Genilloud, Olga; Vicente, Francisca

    2017-01-01

    Ramoplanin is a glycolipodepsipeptide antibiotic obtained from fermentation of Actinoplanes sp. ATCC 33076 that exhibits activity against clinically important multi-drug-resistant, Gram-positive pathogens including vancomycin-resistant Enterococcus (VRE), methicillin-resistant Staphylococcus aureus (MRSA), and vancomycin-intermediate resistant Clostridium difficile. It disrupts bacterial cell wall through a unique mechanism of action by sequestering the peptidoglycan intermediate Lipid II and therefore does not show cross-resistance with other antibiotics. However, while demonstrating excellent antimicrobial activity in systemic use in animal models of infection, ramoplanin presents low local tolerability when injected intravenously. As a consequence of this limitation, new derivatives are desirable to overcome this issue. During a natural product screening program developed to discover compounds that disrupt bacterial cell wall synthesis by inhibiting peptidoglycan transglycosylation through binding to the intermediate Lipid II, 49 actinomycete strains were identified by HR-LCMS as producers of ramoplanin-related compounds. The producing strains were isolated from environmental samples collected worldwide comprising both tropical and temperate areas. To assess the diversity of this microbial population, the 49 isolates were initially identified to the genus level on the basis of their micromorphology, and 16S sequencing confirmed the initial identification of the strains. These analyses resulted in the identification of members of genus Streptomyces, as well as representatives of the families Micromonosporaceae, Nocardiaceae, Thermomonosporaceae, and Pseudonocardiaceae, suggesting that the production of ramoplanins is relatively widespread among Actinomycetes. In addition, all of these isolates were tested against a panel of Gram-positive and Gram-negative bacteria, filamentous fungi, and yeast in order to further characterize their antimicrobial properties. This

  2. Diversity and evolution of secondary metabolism in the marine actinomycete genus Salinispora

    National Research Council Canada - National Science Library

    Nadine Ziemert; Anna Lechner; Matthias Wietz; Natalie Millán-Aguiñaga; Krystle L. Chavarria; Paul Robert Jensen

    2014-01-01

    .... Here we analyze genome sequence data derived from 75 strains of the marine actinomycete genus Salinispora for pathways associated with polyketide and nonribosomal peptide biosynthesis, the products...

  3. [Isolation and antimicrobial activities of actinomycetes from vermicompost].

    Science.gov (United States)

    Wang, Xue-jun; Yan, Shuang-lin; Min, Chang-li; Yang, Yan

    2015-02-01

    In this paper, actinomycetes were isolated from vermicompost by tablet coating method. Antimicrobial activities of actinomycetes were measured by the agar block method. Strains with high activity were identified based on morphology and biochemical characteristics, as well as 16S rDNA gene sequence analysis. The results showed that 26 strains of actinomycetes were isolated, 16 of them had antimicrobial activities to the test strains which accounts for 61.54% of all strains. Among the 16 strains, the strain QYF12 and QYF22 had higher antimicrobial activity to Micrococcus luteus, with a formed inhibition zone of 27 mm and 31 mm, respectively. While the strain QYF26 had higher antimicrobial activity to Bacillus subtilis, and the inhibition zone diameter was 21 mm. Based on the identification of strains with high activity, the strain QYF12 was identified as Streptomyces chartreusis, the strain QYF22 was S. ossamyceticus and the strain QYF26 was S. gancidicus. This study provided a theoretical basis for further separate antibacterial product used for biological control.

  4. ISOLATION AND ANTIMICROBIAL AND DEGRADATIVE POTENTIAL OF ACTINOMYCETES

    Directory of Open Access Journals (Sweden)

    Padma Singh* and Vani Sharma

    2013-02-01

    Full Text Available Problem Statement: Does the soil Actinomycetes have Antimicrobial and Petrol degradation potential? It is an intriguing question. Actinomycetes are continues to be a subject of study with reference to their Antimicrobial and degradative potential. However studies have been done is limited. Our object was to study its Antimicrobial activity in wide spectrum and to study its degradation potential on Petrol. Approach: In this study we have isolated total 5 Actinomycetes from the Ganga river bed. All the isolates later purified and identified by various Morphological and Biochemical test. Here Nocardia was subjected to antimicrobial test against Streptococcus, Mucor and Aspergillus and it was also subjected to degradation test against Petrol. Result: The 5 isolates are Streptomyces, Micromonospora, Micromono sporangium and 2 different strain of Nocardia (Na1 and Na2. The 2 strains of Nocardia are active against Streptococcus (Na1 29.6mm, Na2 26.6mm, Mucor (Na1 12.5mm, Na2 22.5mm and Aspergillus (Na1 50%, Na2 60%. They also degrade Petrol very effectively, decrease in total organic carbon of the medium was observed during the degradation of petrol. Conclusion: Our observation provides us with evidence that these agents can be used for the production of new antibiotics and as the agent to control the environment pollution.

  5. CRISPR-Cas9 Based Engineering of Actinomycetal Genomes.

    Science.gov (United States)

    Tong, Yaojun; Charusanti, Pep; Zhang, Lixin; Weber, Tilmann; Lee, Sang Yup

    2015-09-18

    Bacteria of the order Actinomycetales are one of the most important sources of pharmacologically active and industrially relevant secondary metabolites. Unfortunately, many of them are still recalcitrant to genetic manipulation, which is a bottleneck for systematic metabolic engineering. To facilitate the genetic manipulation of actinomycetes, we developed a highly efficient CRISPR-Cas9 system to delete gene(s) or gene cluster(s), implement precise gene replacements, and reversibly control gene expression in actinomycetes. We demonstrate our system by targeting two genes, actIORF1 (SCO5087) and actVB (SCO5092), from the actinorhodin biosynthetic gene cluster in Streptomyces coelicolor A3(2). Our CRISPR-Cas9 system successfully inactivated the targeted genes. When no templates for homology-directed repair (HDR) were present, the site-specific DNA double-strand breaks (DSBs) introduced by Cas9 were repaired through the error-prone nonhomologous end joining (NHEJ) pathway, resulting in a library of deletions with variable sizes around the targeted sequence. If templates for HDR were provided at the same time, precise deletions of the targeted gene were observed with near 100% frequency. Moreover, we developed a system to efficiently and reversibly control expression of target genes, deemed CRISPRi, based on a catalytically dead variant of Cas9 (dCas9). The CRISPR-Cas9 based system described here comprises a powerful and broadly applicable set of tools to manipulate actinomycetal genomes.

  6. Airway inflammation among compost workers exposed to actinomycetes spores

    Directory of Open Access Journals (Sweden)

    Kari Kulvik Heldal

    2015-05-01

    Full Text Available Objectives. To study the associations between exposure to bioaerosols and work-related symptoms, lung function and biomarkers of airway inflammation in compost workers. Materials and method. Personal full-shift exposure measurements were performed on 47 workers employed at five windrow plants (n=20 and five reactor plants (n=27. Samples were analyzed for endotoxins, bacteria, fungal and actinomycetes spores. Health examinations were performed on workers and 37 controls before and after work on the day exposure was measured. The examinations included symptoms recorded by questionnaire, lung function by spirometry and nasal dimensions by acoustic rhinometry (AR. The pneumoproteins CC16, SP-D and SP-A were measured in a blood sample drawn at the end of the day. Results. The levels of endotoxins (median 3 EU/m[sup]3[/sup] , range 0–730 EU/m[sup]3[/sup] and actinomycetes spores (median 0.2 × 10[sup]6[/sup] spores/m[sup]3[/sup] , range 0–590 × 10[sup]6[/sup] spores/m[sup]3[/sup] were significantly higher in reactor plants compared to windrow plants. However, windrow composting workers reported more symptoms than reactor composting workers, probably due to use of respiratory protection. Exposure-response relationships between actinomycetes spores exposure and respiratory effects, found as cough and nose irritation during a shift, was significantly increased (OR 4.3, 95% CI 1.1–16, OR 6.1, 95% CI 1.5–25, respectively, p<0.05 among workers exposed to 0.02–0.3 × 10[sup]6[/sup] actinomycetes spores/m 3 , and FEV1/FVC% decreased cross shift (b=–3.2, SE=1.5%, p<0.01. Effects were weaker in the highest exposed group, but these workers used respiratory protection, frequently limiting their actual exposure. No relationships were found between exposure and pneumoprotein concentrations. Conclusions. The major agent in the aerosol generated at compost plants was actinomycetes spores which was associated with work related cough symptoms and work

  7. Heterologous expression of Thermobifida fusca thermostable alpha-amylase in Yarrowia lipolytica and its application in boiling stable resistant sago starch preparation.

    Science.gov (United States)

    Yang, Chao-Hsun; Huang, Yu-Chun; Chen, Cheng-Yu; Wen, Chia-Ying

    2010-09-01

    A gene encoding the thermostable alpha-amylase in Thermobifida fusca NTU22 was amplified by PCR, sequenced, and cloned into Yarrowia lipolytica P01g host strain using the vector pYLSC1 allowing constitutive expression and secretion of the protein. Recombinant expression resulted in high levels of extracellular amylase production, as high as 730 U/l in the Hinton flask culture broth. It is higher than that observed in P. pastoris expression system and E. coli expression system. The purified amylase showed a single band at about 65 kDa by SDS-polyacrylamide gel electrophoresis and this agrees with the predicted size based on the nucleotide sequence. About 70% of the original activity remained after heat treatment at 60 degrees C for 3 h. The optimal pH and temperature of the purified amylase were 7.0 and 60 degrees C, respectively. The purified amylase exhibited a high level of activity with raw sago starch. After 72-h treatment, the DP(w) of raw sago starch obviously decreased from 830,945 to 237,092. The boiling stable resistant starch content of the sago starch increased from 8.3 to 18.1%. The starch recovery rate was 71%.

  8. Expression of Thermobifida fusca thermostable raw starch digesting alpha-amylase in Pichia pastoris and its application in raw sago starch hydrolysis.

    Science.gov (United States)

    Yang, Chao-Hsun; Huang, Yu-Chun; Chen, Cheng-Yu; Wen, Chia-Ying

    2010-04-01

    A gene encoding the thermostable raw starch digesting alpha-amylase in Thermobifida fusca NTU22 was amplified by PCR, sequenced and cloned into Pichia pastoris X-33 host strain using the vector pGAPZalphaA, allowing constitutive expression and secretion of the protein. Recombinant expression resulted in high levels of extracellular amylase production, as high as 510 U/l in the Hinton flask culture broth. The purified amylase showed a single band at about 65 kDa by SDS-polyacrylamide gel electrophoresis after being treated with endo-beta-N-acetylglycosaminidase H, and this agrees with the predicted size based on the nucleotide sequence. About 75% of the original activity remained after heat treatment at 60 degrees C for 3 h. The optimal pH and temperature of the purified amylase were 7.0 and 60 degrees C, respectively. The purified amylase exhibited a high level of activity with raw sago starch. After 48-h treatment, the DPw of raw sago starch obviously decreased from 830,945 to 378,732. The surface of starch granules was rough, and some granules displayed deep cavities.

  9. Cellulolytic and proteolytic ability of bacteria isolated from gastrointestinal tract and composting of a hippopotamus.

    Science.gov (United States)

    da Cruz Ramos, Geomárcia Feitosa; Ramos, Patricia Locosque; Passarini, Michel Rodrigo Zambrano; Vieira Silveira, Marghuel A; Okamoto, Débora Noma; de Oliveira, Lilian Caroline Gonçalves; Zezzo, Larissa Vieira; Marem, Alyne; Santos Rocha, Rafael Costa; da Cruz, João Batista; Juliano, Luiz; de Vasconcellos, Suzan Pantaroto

    2016-03-01

    The bioprospection for cellulase and protease producers is a promise strategy for the discovery of potential biocatalysts for use in hydrolysis of lignocellulosic materials as well as proteic residues. These enzymes can increment and turn viable the production of second generation ethanol from different and alternative sources. In this context, the goal of this study was the investigation of cellulolytic and proteolytic abilities of bacteria isolated from the gastrointestinal tract of a hippopotamus as well as from its composting process. It is important to highlight that hippopotamus gastrointestinal samples were a non-typical sources of efficient hydrolytic bacteria with potential for application in biotechnological industries, like biofuel production. Looking for this, a total of 159 bacteria were isolated, which were submitted to qualitative and quantitative enzymatic assays. Proteolytic analyzes were conducted through the evaluation of fluorescent probes. Qualitative assays for cellulolytic abilities revealed 70 positive hits. After quantitative analyzes, 44 % of these positive hits were selected, but five (5) strains showed cellulolytic activity up to 11,8 FPU/mL. Regarding to proteolytic activities, six (6) strains showed activity above 10 %, which overpassed results described in the literature. Molecular analyzes based on the identification of 16S rDNA, revealed that all the selected bacterial isolates were affiliated to Bacillus genus. In summary, these results strongly indicate that the isolated bacteria from a hippopotamus can be a potential source of interesting biocatalysts with cellulolytic and proteolytic activities, with relevance for industrial applications.

  10. Sample handling factors affecting the enumeration of lactobacilli and cellulolytic bacteria in equine feces

    Science.gov (United States)

    The objectives were to compare media types and evaluate the effects of fecal storage time and temperature on the enumeration of cellulolytic bacteria and lactobacilli from horses. Fecal samples were collected from horses (n = 3) and transported to the lab (CO2, 37 ºC, 0.5 h). The samples were assign...

  11. Recombinant host cells and nucleic acid constructs encoding polypeptides having cellulolytic enhancing activity

    Energy Technology Data Exchange (ETDEWEB)

    Schnorr, Kirk; Kramer, Randall

    2017-03-28

    The present invention relates to isolated polypeptides having cellulolytic enhancing activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

  12. Determination of the cellulolytic activities of microorganisms isolated from poultry litter for sawdust degradation

    Directory of Open Access Journals (Sweden)

    Akpomie O.OF

    2013-03-01

    Full Text Available Cellulolytic activities of bacterial and fungal isolates obtained from poultry droppings were determined using the ability of each isolate to produce clear zones on Carboxyl Methyl Cellulose Agar plates. The bacterial isolates were Klebsiella, Streptococcus, Celulomonas, Escherichia coli and Micrococus species. The cellulolytic counts ranged from 5.02 x 104 + 3.42 to 7.20 x 109 + 6.12 cfu/g. The cellulolytic activities of the bacterial isolates ranged from 0.04 to 0.26 iu/m with Cellulomonas having the highest cellulose activity. The fungal isolates were Aspergillus niger, Mucor mucedo, Trichoderma sp. and Penicllium chrysogenum with cellulose activities of 0.24 + 0.021 0.19 + 0.031, 0.23 + 0.05 and 0.23 + 0.028iu/ml respectively. All the isolates were able to degrade the sawdust to varying extent. The percentage degradation was highest with Micrococcus sp. (78.20% and least with Trichoderma sp. (65.83%. The study shows that is a potential source of cellulolytic microorganisms which could be employed in the degradation of sawdust.

  13. [Biosynthesis of cellulolytic enzymes and xylanase during submerged cultivation of the fungus Aspergillus terreus 17P].

    Science.gov (United States)

    Loginova, L G; Guzhova, E P; Ismanlova, D Iu; Burdenko, L G

    1978-01-01

    The fungus Aspergillus terreus 17P--producer of cellulolytic enzymes--was cultivated in the Biotec 10 l fermenter on the medium containing minced and heated (at 200 degrees) wheat straw aerated with a different rate. At the mixing rate of 350 rpm and aeration rate of 0.7 r/rpm on the fourth day the culture liquid was obtained whose filtrate contained an active complex of cellulolytic enzymes and xylanase: CI--3.4; APB--1.1, Cx--35.7, cellobiase--0.23, xylanase--73.8 units/ml. The fractionation of the culture liquid filtrate with ammonium sulphate showed that the fraction precipitated at an interval of saturation of 0.3--0.7 contained the largest portion of cellulolytic enzymes and xylanase. The isolated enzymic preparations had a cellulolytic and xylanase activity and contained lipase, pectinase, laminarinase. They also contained low quantities of amylase, protease, beta-1,4- and beta-1,6-glucanase. Enzymic hylrolysis by the Asp. terreus 17P preparation of straw yielded glucose and xylose, of cotton, Na-KMC, cellobiose--glucose, Xylane hydrolyzate contained xylose and arabinose.

  14. Construction and Characterization of a Cellulolytic Consortium Enriched from the Hindgut of Holotrichia parallela Larvae

    Directory of Open Access Journals (Sweden)

    Ping Sheng

    2016-09-01

    Full Text Available Degradation of rice straw by cooperative microbial activities is at present the most attractive alternative to fuels and provides a basis for biomass conversion. The use of microbial consortia in the biodegradation of lignocelluloses could reduce problems such as incomplete synergistic enzymes, end-product inhibition, and so on. In this study, a cellulolytic microbial consortium was enriched from the hindgut of Holotrichia parallela larvae via continuous subcultivation (20 subcultures in total under static conditions. The degradation ratio for rice straw was about 83.1% after three days of cultivation, indicating its strong cellulolytic activity. The diversity analysis results showed that the bacterial diversity and richness decreased during the consortium enrichment process, and the consortium enrichment process could lead to a significant enrichment of phyla Proteobacteria and Spirochaetes, classes Clostridia, Epsilonproteobacteria, and Betaproteobacteria, and genera Arcobacter, Treponema, Comamonas, and Clostridium. Some of these are well known as typical cellulolytic and hemicellulolytic microorganisms. Our results revealed that the microbial consortium identified herein is a potential candidate for use in the degradation of waste lignocellulosic biomass and further highlights the hindgut of the larvae as a reservoir of extensive and specific cellulolytic and hemicellulolytic microbes.

  15. Construction and Characterization of a Cellulolytic Consortium Enriched from the Hindgut of Holotrichia parallela Larvae

    Science.gov (United States)

    Sheng, Ping; Huang, Jiangli; Zhang, Zhihong; Wang, Dongsheng; Tian, Xiaojuan; Ding, Jiannan

    2016-01-01

    Degradation of rice straw by cooperative microbial activities is at present the most attractive alternative to fuels and provides a basis for biomass conversion. The use of microbial consortia in the biodegradation of lignocelluloses could reduce problems such as incomplete synergistic enzymes, end-product inhibition, and so on. In this study, a cellulolytic microbial consortium was enriched from the hindgut of Holotrichia parallela larvae via continuous subcultivation (20 subcultures in total) under static conditions. The degradation ratio for rice straw was about 83.1% after three days of cultivation, indicating its strong cellulolytic activity. The diversity analysis results showed that the bacterial diversity and richness decreased during the consortium enrichment process, and the consortium enrichment process could lead to a significant enrichment of phyla Proteobacteria and Spirochaetes, classes Clostridia, Epsilonproteobacteria, and Betaproteobacteria, and genera Arcobacter, Treponema, Comamonas, and Clostridium. Some of these are well known as typical cellulolytic and hemicellulolytic microorganisms. Our results revealed that the microbial consortium identified herein is a potential candidate for use in the degradation of waste lignocellulosic biomass and further highlights the hindgut of the larvae as a reservoir of extensive and specific cellulolytic and hemicellulolytic microbes. PMID:27706065

  16. [Date palm and fusariosis. VIII.--Parasitism of "Fusarium oxysporum" f. sp. "albedinis" by an actinomycete (author's transl)].

    Science.gov (United States)

    Sabaou, N; Bennaceur, M; Bounaga, D

    1981-01-01

    Fortuitous growth of an actinomycete on Fusarium oxysporum f. sp. albedinis culture has shown a host-parasite process. As a response to the actinomycete, the fungus produces thallospores with various forms which can germinate faster than the non-parasited F. o. albedinis microconidies. However, the strains obtained from thallospores showed as sensible as the mother strain towards actinomycete action.

  17. Marine actinomycetes from Madeira Archipelago preliminary taxonomic studies

    Directory of Open Access Journals (Sweden)

    Ilda Santos Sanches

    2014-06-01

    Full Text Available The oceans cover 70 % of the Earth´s surface and harbor most of the planet´s biodiversity. However the microbiological component of this diversity remains relatively unexplored. Marine actinomycetes, are a robust resource of chemically prolific novelty. Producing structurally unique biological active secondary metabolites, generating a valuable source for innovative biotechnology and drug discovery[1,2]. As a consequence, the ecological role of actinomycetes and their marine ecosystems may no longer be neglected. It is crucial to move our research efforts into ocean regions for which we know little or nothing about the indigenous microbial diversity. The Portuguese Archipelago, Madeira is located in the Macaronesian Atlantic region, emerging from the African tectonic plate, found in the extreme south of the Tore-Madeira ridge, has a unique biogeography and biodiversity. These distinctive characteristics combined with the fact that Madeira have never been explored, as far as indigenous marine actinomycetes are concerned, makes it from the scientific point of view, the perfect target for our studies. From 662 marine sediment samples collected along Madeira Archipelago (Figure 1 during June of 2012, covering depths from 10-1310 m, a total of 421 actinomycete strains were isolated. In a previous study, an assemblage of 82 strains was selected for taxonomic identification, having into account representative morphological diversity characteristics of the actinomycetes, isolated from Madeira Archipelago. Based on 16S rRNA gene sequencing, it was observed that the genera Streptomyces, Micromonospora and Salinispora were predominant, 81% [3]. Additionally, in a recent study, our team selected 168 strains with Salinispora look-alike morphological features. From these 28 strains were identified as belonging to the seawater-obligate marine actinomycete genus Salinispora. Representing the first report of Salinispora spp. in the Macaronesian Atlantic Ocean

  18. Actinomycete integrative and conjugative pMEA-like elements of Amycolatopsis and Saccharopolyspora decoded

    NARCIS (Netherlands)

    Poele, Evelien M. Te; Samborskyy, Markiyan; Oliynyk, Markiyan; Leadlay, Peter F.; Bolhuis, Henk; Dijkhuizen, Lubbert

    Actinomycete integrative and conjugative elements (AICEs) are present in diverse genera of the actinomycetes, the most important bacterial producers of bioactive secondary metabolites. Comparison of pMEA100 of Amycolatopsis mediterranei, pMEA300 of Amycolatopsis methanolica and pSE211 of

  19. [Secondary Metabolites from Marine Microorganisms. I. Secondary Metabolites from Marine Actinomycetes].

    Science.gov (United States)

    Orlova, T I; Bulgakova, V G; Polin, A N

    2015-01-01

    Review represents data on new active metabolites isolated from marine actinomycetes published in 2007 to 2014. Marine actinomycetes are an unlimited source of novel secondary metabolites with various biological activities. Among them there are antibiotics, anticancer compounds, inhibitors of biochemical processes.

  20. Actinomycete integrative and conjugative pMEA-like elements of Amycolatopsis and Saccharopolyspora decoded

    NARCIS (Netherlands)

    Te Poele, E.M.; Samborskyy, M.; Oliynyk, M.; Leadlay, P.F.; Bolhuis, H.; Dijkhuizen, L.

    2008-01-01

    Actinomycete integrative and conjugative elements (AICEs) are present in diverse genera of the actinomycetes, the most important bacterial producers of bioactive secondary metabolites. Comparison of pMEA100 of Amycolatopsis mediterranei, pMEA300 of Amycolatopsis methanolica and pSE211 of

  1. In Vitro Investigation of Antifungal Activities of Actinomycetes against Microsporum gypseum

    Directory of Open Access Journals (Sweden)

    Naser Keikha

    2013-02-01

    Conclusion: The findings of the present research show that terrigenous actinomycetes have an antifungal effect upon Microsporum gypseum. So, one hopes that-in future-rather than administering antifungal chemicals that have side-effects, dermatophytic infections can be cured by applying these actinomycetes.

  2. Actinomycete integrative and conjugative pMEA-like elements of Amycolatopsis and Saccharopolyspora decoded

    NARCIS (Netherlands)

    Poele, Evelien M. Te; Samborskyy, Markiyan; Oliynyk, Markiyan; Leadlay, Peter F.; Bolhuis, Henk; Dijkhuizen, Lubbert

    2008-01-01

    Actinomycete integrative and conjugative elements (AICEs) are present in diverse genera of the actinomycetes, the most important bacterial producers of bioactive secondary metabolites. Comparison of pMEA100 of Amycolatopsis mediterranei, pMEA300 of Amycolatopsis methanolica and pSE211 of Saccharopol

  3. [Number and structure of actinomycetes complexes in the rhizosphere winter rye, oat and red clover].

    Science.gov (United States)

    Shirokikh, I G; Zenova, G M; Merzaeva, O V; Lapygina, E V; Lysak, L V

    2006-01-01

    The actinomycetes complexes in the rhizosphere of three agricultural plants by using the methods of luminescense microscopy and cup sowing were investigated. It was established, that concentration of prokaryotic biomass and biomass of actinomycetes mycelium in rhizosphere of plants is higher than in free from the radicals to soil. Rhizosphera of the oat (Avena sativa L.) and red clover (Trifolium pratense L.) is colonized by Streptomyces, Micromonospora and olygospore species. Dominante actinomycetes of winter rye (Secale cereale L.) are classified into the genera Micromonospora. It was shown that numbers and biomass of actinomycetes mycelium were fond to decreased, diversity of actinomycetes in contrast is increased in the series: "winter rye--oat--red clover". In connection with ecological safety the capability of increase with prokaryotes naturally disease suppressive soil and stability of plants to pathogen is discussed.

  4. New Benzoxazine Secondary Metabolites from an Arctic Actinomycete

    OpenAIRE

    Kyuho Moon; Chan-Hong Ahn; Yoonho Shin; Tae Hyung Won; Keebeom Ko; Sang Kook Lee; Ki-Bong Oh; Jongheon Shin; Seung-Il Nam; Dong-Chan Oh

    2014-01-01

    Two new secondary metabolites, arcticoside (1) and C-1027 chromophore-V (2), were isolated along with C-1027 chromophore-III and fijiolides A and B (3–5) from a culture of an Arctic marine actinomycete Streptomyces strain. The chemical structures of 1 and 2 were elucidated through NMR, mass, UV, and IR spectroscopy. The hexose moieties in 1 were determined to be d-glucose from a combination of acid hydrolysis, derivatization, and gas chromatographic analyses. Arcticoside (1) and C-1027 chromo...

  5. Therapeutic Potential of Biologically Reduced Silver Nanoparticles from Actinomycete Cultures

    Directory of Open Access Journals (Sweden)

    M. K. Sukanya

    2013-01-01

    Full Text Available Silver nanoparticles are applied in nanomedicine from time immemorial and are still used as powerful antibiotic and anti-inflammatory agents. Antibiotics produced by actinomycetes are popular in almost all the therapeutic measures, and this study has proven that these microbes are also helpful in the biosynthesis of silver nanoparticles with good surface and size characteristics. Silver can be synthesized by various chemical methodologies, and most of them have turned to be toxic. This study has been successful in isolating the microbes from polluted environment, and subjecting them to the reduction of silver nanoparticles, characterizing the nanoparticles by UV spectrophotometry and transmission electron microscopy. The nanoparticles produced were tested for their antimicrobial property, and the zone of inhibition was greater than those produced by their chemically synthesized counterparts. Actinomycetes, helpful in bioremediating heavy metals, are useful for the production of metallic nanoparticles. The biosynthesized silver nanoparticles loaded with antibiotics prove to be better in killing the pathogens and have opened up new areas for developing nanobiotechnological research based on microbial applications.

  6. New Dimensions of Research on Actinomycetes: Quest for Next Generation Antibiotics

    Directory of Open Access Journals (Sweden)

    Polpass Arul Jose

    2016-08-01

    Full Text Available Starting with the discovery of streptomycin, the promise of natural products research on actinomycetes has been captivat¬ing researchers and offered an array of life-saving antibiotics. However, most of the actinomycetes have received a little attention of researchers beyond isolation and activity screening. Noticeable gaps in genomic information and associated biosynthetic potential of actinomycetes are mainly the reasons for this situation, which has led to a decline in the discovery rate of novel antibiotics. Recent insights gained from genome mining have revealed a massive existence of previously unrecognized biosynthetic potential in actinomycetes. Successive developments in next-generation sequencing, genome editing, analytical separation and high-resolution spectroscopic methods have reinvigorated interest on such actinomycetes and opened new avenues for the discovery of natural and natural-inspired antibiotics. This article describes the new dimensions that have driven the ongoing resurgence of research on actinomycetes with historical background since the commencement in 1940, for the attention of worldwide researchers. Coupled with increasing advancement in molecular and analytical tools and techniques, the discovery of next-generation antibiotics could be possible by revisiting the untapped potential of actinomycetes from different natural sources.

  7. Bioprospecting marine actinomycetes for multidrug-resistant pathogen control from Rameswaram coastal area, Tamil Nadu, India.

    Science.gov (United States)

    Wahaab, Femina; Subramaniam, Kalidass

    2017-08-07

    A potent Streptomyces bacillaris strain RAM25C4 was isolated for controlling methicillin-resistant Staphylococcus aureus and multidrug-resistant bacteria such as Staphylococcus aureus, Acinetobacter baumannii, and Pseudomonas aeruginosa. A total of 131 actinomycetes were isolated from the Rameswaram coastal region, Tamil Nadu, India. Among 131 actinomycetes, maximum number of actinomycetes (55%) isolated at the distance of 3-6 m from seashore. Out of 131 actinomycetes, 85% of the actinomycetes exhibited different degree of antagonistic activity against test pathogens. The antagonistic activity evaluated using actinomycetes direct culture filtrate and culture filtrate extracts. Among these culture filtrate, extracts had supreme antagonistic activity against multidrug-resistant bacteria and the solvent ethyl acetate was the best for extracting secondary metabolites from actinomycetes. In HPTLC analysis, the presence of macrolides, terpenoids, and quinolones was identified in RAM25C4 extract. In GC-MS analysis, various potent compounds such as phenolic compound-2,6-di-tert-butylphenol, alkaloid compound-1H, 5H, pyrrolo (1' 2':3, 4) imidazo, and quinolone compound-1,4-benzenediol, 2,5-bis(1,1-dimethylethyl) were identified in the ethyl acetate extract of RAM25C4. The phylogenetic analysis of 16S rRNA gene sequence of RAM25C4 isolate was deposited in NCBI with name Streptomyces bacillaris strain RAM25C4 and accession number KM513543.

  8. Diversity and Antagonistic Activity of Actinomycete Strains From Myristica Swamp Soils Against Human Pathogens

    Directory of Open Access Journals (Sweden)

    Varghese Rlnoy

    2014-05-01

    Full Text Available Under the present investigation Actinomycetes were isolated from the soils of Myristica swamps of southern Western Ghats and the antagonistic activity against different human bacterial pathogens was evaluated. Results of the present study revealed that Actinomycetes population in the soils of Myristica swamp was spatially and seasonally varied. Actinomycetes load was varied from 24×104 to 71×103, from 129×103 to 40×103 and from 31×104 to 84×103 in post monsoon, monsoon and pre monsoon respectively. A total of 23 Actinomycetes strains belonging to six genera were isolated from swamp soils. Identification of the isolates showed that most of the isolates belonged to the genus Streptomyces (11, followed by Nocardia (6, Micromonospora (3, Pseudonocardia (1, Streptosporangium (1, and Nocardiopsis (1. Antagonistic studies revealed that 91.3% of Actinomycete isolates were active against one or more tested pathogens, of that 56.52% exhibited activity against Gram negative and 86.95% showed activity against Gram positive bacteria. 39.13% isolates were active against all the bacterial pathogens selected and its inhibition zone diameter was also high. 69.5% of Actinomycetes were exhibited antibacterial activity against Listeria followed by Bacillus cereus (65.21%, Staphylococcus (60.86%, Vibrio cholera (52.17%, Salmonella (52.17% and E. coli (39.13%. The results indicate that the Myristica swamp soils of Southern Western Ghats might be a remarkable reserve of Actinomycetes with potential antagonistic activity.

  9. [Phylogenetic diversity of the culturable rare actinomycetes in marine sponge Hymeniacidon perlevis by improved isolation media].

    Science.gov (United States)

    Xin, Yanjuan; Wu, Peichun; Deng, Maicun; Zhang, Wei

    2009-07-01

    Based on the molecular diversity information, seven actinomycete-selective culture media and isolation conditions were modified to isolate and cultivate diverse rare actinomycetes from Hymeniacidon perlevis. Modified, selective cultivation and enrichment media were used, with the addition of an elemental solution of simulating the elemental composition of marine sponge H. perlevis. Restriction Fragment Length Polymorphism (RFLP) analysis of 16S rDNA sequence was used to reveal the diversity of culturable rare actinomycetes. A total of 59 actinomycete strains were isolated from the marine sponge H. perlevis. A total of 27 representative actinomycetes were selected according to their morphological feature, color and pigments. They gave 15 different RFLP patterns after digesting their PCR products of 16s rDNA with Hha I. The results showed that these isolates belonged to 10 genera: Streptomyces, Nocardiopsis, Micromonospora, Cellulosimicrobium, Gordonia, Nocardia, Prauseria, Pseudonocardia , Saccharomonospora and Microbacterium. The modified isolation media and selective cultivation procedures are highly effective in the recovery of culturable actinomycetes from the marine sponge H. perlevis, resulting in the highest diversity of culturable rare actinomycetes from any sponges.

  10. Screening of Actinomycete Isolates from Niche Habitats in Manipur for Antibiotic Activity

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    Debananda S. Ningthoujam

    2009-01-01

    Full Text Available Problem statement: The exhaustion of the usual terrestrial sources and the rise of resistant pathogens dictate the search for novel actinomycetes and new antibiotics. In this context, niche habitats such as caves, pristine forests, lakes, rivers, and other wetlands, high salt environments, marine ecosystems and endophytic niches are promising targets for survey of bioactive actinomycetes. Approach: Actinomycetes were isolated from several niche habitats in Manipur, India, on selective media such as SCNA and Chitin agar with or without antibiotics. Selected isolates were subjected to antimicrobial activity screening by Kirby-Bauer method. Results: 172 lake sediment (SCNA, LS1 series, 35 lake sediment (CA, LSCH series, 120 river (NRP, NRB and..series, 39 forest (AML series, 35 cave (KC1 series, 101 salt spring (NH, N3S and .. series, 46 Shirui jungle (SJ series and 66 Shirui hill (SH series actinomycetes isolates were obtained. Of 99 randomly selected isolates screened, 37 had antimicrobial activities against 1 or more indicator strains: 32 against Gram positive bacteria and 8 against Gram negative bacteria; 10 actinomycete strains were antimycotic and 3 had broad-spectrum antibiotic activities. About 18 potent antibacterial, 1 anti pseudomonas, 1 exclusively antifungal and 3 broad-spectrum antimicrobial actinomycetes were chosen for further studies. Conclusion: Niche habitats in Manipur especially wetlands show great promise for discovery of bioactive actinomycetes.

  11. Terrestrial actinomycetes from diverse locations of Uttarakhnad, India: Isolation and screening for their antibacterial activity.

    Directory of Open Access Journals (Sweden)

    Vijay Kumar

    2013-09-01

    Full Text Available Uttarakhand region is less explored, but possess a great biodiversity. This diversity can be explored for isolation and characterization of new actinomycetes strains for seeking antimicrobial molecules. It can therefore be predicted that novel bioactive metabolite producing actinomycetes can be discovered to combat multidrug resistant bacterial pathogens.Variations in the viable count of actinomycetes were accessed in different altitudes. Actinomycetes were isolated, indentified and screened for their antibacterial activity.The highest viable counts of actinomycetes were recorded in valleys followed by mid hills and high hills. A total of 512 actinomycetes were isolated which were found to belong the 14 different genera of actinomycetes. Mainly the genus Streptomyces was dominant in all the soil samples. Out of 512 isolates recovered, 23.44% exhibited antibacterial activity against one or more tested bacterial pathogens. Of these 56.67% showed activity against Gram-positive bacteria, 26.67% against Gram-negative bacteria while 16.67% showed broad spectrum activity. Isolate DV1S and GR9a-5 showed highest antibacterial properties against several multi-drug resistant bacterial pathogens and were identified using polyphasic approach. DV1S and GR9a-5 were found to be most closely related with S. massasporeus NBRC 12796(T and Nocardia nova JCM 6044(T respectively.The results of this study strongly support the idea that the viable count of actinomycetes varied greatly with altitude. The actinomycetes species isolated from valleys, mid hills and high hills possess significant capacity to produce compounds which are active against several drug resistant bacterial pathogens.

  12. Studies on Actinomycetal Resources under Extreme Environments in the West of China

    Science.gov (United States)

    Li, W.

    2005-12-01

    s: Actinomycetes play a quite important role in natural ecological system and they are also profile producers of antibiotics, antitumor agents, enzymes, enzyme inhibitors and immunomodifiers. which have been widely applied in industry, agriculture, forestry and pharmaceutical industry. In the past, the research work on actinomycetes was mainly concentrated on that of common habitats. Actinomycetes resources under extreme environments (including extreme high and low temperature, extreme high or low pH, high salt concentration etc.) have received comparatively little attention from microbiologists. Actinomycetes are regarded as one kind of sideline microorganisms and those under extreme environments are better materials for biological evolution and phylogenetic development in research. There are much more unknown species and much more worth researching for actinomycetes under extreme environments. There are many extreme environmental resources in the west of China. For example, wide range snow-mountains, basified soil and lakes, widely distributed acid and alkaline hot-springs in Yunnan provinces; more than 73.3 million hektares basified soil and salt lakes in Xinjiang Province and many unusual environments in Qinghai Province and other western Provinces. They were mostly precious natural resources and were destroyed, relatively fewer can provided us with unique conditions for study on actinomycetal resources under extreme environments. In recent years, our main work was focusing on study of extremophilic actinomycetal resources in the west of China by using conventional cultivation-methods and culture-independent methods (PCR-clone and DGGE/TGGE, etc), Results showed that large amount of unknown microbial resources (including actinomycetal resources) existed in natural extreme environments. Additionally, lots of new taxa were isolated and characterized using a polyphasic approach. Further, we got some new compounds with different bioactivities from these

  13. A potential source for cellulolytic enzyme discovery and environmental aspects revealed through metagenomics of Brazilian mangroves

    OpenAIRE

    Thompson, Claudia Elizabeth; Silva, Walter Orlando Beys da; Santi, Lucélia; Oliveira, Markus Berger; Vainstein, Marilene Henning; Guimaraes, Jorge Almeida; Vasconcelos, Ana Tereza Ribeiro

    2013-01-01

    The mangroves are among the most productive and biologically important environments. The possible presence of cellulolytic enzymes and microorganisms useful for biomass degradation as well as taxonomic and functional aspects of two Brazilian mangroves were evaluated using cultivation and metagenomic approaches. From a total of 296 microorganisms with visual differences in colony morphology and growth (including bacteria, yeast and filamentous fungus), 179 (60.5%) and 117 (39.5%) were isolated...

  14. Cellulolytic and hemicellulolytic bacteria from the gut of Oryctes rhinoceros larvae

    Directory of Open Access Journals (Sweden)

    SITI LUSI ARUM SARI

    2016-04-01

    Full Text Available Abstract. Sari SLA, Pangstuti A, Susilowati A, Purwoko Tj, Mahajoeno E, Hidayat W, Mardhena I, Panuntun DF, Kurniawati D, Anitasari R. 2016. Cellulolytic and hemicellulolytic bacteria from the gut of Oryctes rhinoceros larvae. Biodiversitas 17: 78-83. Lignocellulose is very potential as raw material for biofuel production because it is cheap, abundant and renewable. The main carbohydrate constituents of lignocellulosic material are cellulose and hemicelluloses (a group of heteropolymers that includes xylans and mannans. The most important process in bioethanol production from lignocellulose is the bioconversion of polysacharides into fermentable sugar. Enzymatic hydrolysis has been developed because it is the more environmentally approach. Since the cost of hydrolytic enzyme production is the major problem of the process, many type of research has been focused on lowering the cost of enzyme production, including screening for organisms with a novel enzyme. This present study was conducted to isolate and screen of the cellulolytic and hemicellulolytic Bacteria from the gut of Oryctes rhinoceros L. larvae. The 3rd instars were used in this research.The research succeeded to isolate 11 bacterial isolates from the gut of O. rhinoceros larvae. The screening result demonstrated that bacterial isolates had cellulolytic (63.6% of total isolates, xylanolytic (72.7% of total isolates, and mannanolytic (100% of total isolates activity. Based on the 16S rDNA sequence, 10 isolates were classified into Bacillus and only 1 isolate was classified into Citrobacter. The GOR2 which was closely related to Bacillus pumilus vit bac1 has the highest cellulolytic and xyllanolytic activities. The isolate with the highest mannanolytic activity was the GOR7 which was closely related to Bacillus aryabhattai strain IHB B 6821.

  15. Anaerobic gut fungi: Advances in isolation, culture, and cellulolytic enzyme discovery for biofuel production.

    Science.gov (United States)

    Haitjema, Charles H; Solomon, Kevin V; Henske, John K; Theodorou, Michael K; O'Malley, Michelle A

    2014-08-01

    Anaerobic gut fungi are an early branching family of fungi that are commonly found in the digestive tract of ruminants and monogastric herbivores. It is becoming increasingly clear that they are the primary colonizers of ingested plant biomass, and that they significantly contribute to the decomposition of plant biomass into fermentable sugars. As such, anaerobic fungi harbor a rich reservoir of undiscovered cellulolytic enzymes and enzyme complexes that can potentially transform the conversion of lignocellulose into bioenergy products. Despite their unique evolutionary history and cellulolytic activity, few species have been isolated and studied in great detail. As a result, their life cycle, cellular physiology, genetics, and cellulolytic metabolism remain poorly understood compared to aerobic fungi. To help address this limitation, this review briefly summarizes the current body of knowledge pertaining to anaerobic fungal biology, and describes progress made in the isolation, cultivation, molecular characterization, and long-term preservation of these microbes. We also discuss recent cellulase- and cellulosome-discovery efforts from gut fungi, and how these interesting, non-model microbes could be further adapted for biotechnology applications. © 2014 Wiley Periodicals, Inc.

  16. Use of Cellulolytic Marine Bacteria for Enzymatic Pretreatment in Microalgal Biogas Production

    Science.gov (United States)

    Muñoz, Camilo; Hidalgo, Catalina; Zapata, Manuel; Jeison, David; Riquelme, Carlos

    2014-01-01

    In this study, we designed and evaluated a microalgal pretreatment method using cellulolytic bacteria that naturally degrades microalgae in their native habitat. Bacterial strains were isolated from each of two mollusk species in a medium containing 1% carboxymethyl cellulose agar. We selected nine bacterial strains that had endoglucanase activity: five strains from Mytilus chilensis, a Chilean mussel, and four strains from Mesodesma donacium, a clam found in the Southern Pacific. These strains were identified phylogenetically as belonging to the genera Aeromonas, Pseudomonas, Chryseobacterium, and Raoultella. The cellulase-producing capacities of these strains were characterized, and the degradation of cell walls in Botryococcus braunii and Nannochloropsis gaditana was tested with “whole-cell” cellulolytic experiments. Aeromonas bivalvium MA2, Raoultella ornithinolytica MA5, and Aeromonas salmonicida MC25 degraded B. braunii, and R. ornithinolytica MC3 and MA5 degraded N. gaditana. In addition, N. gaditana was pretreated with R. ornithinolytica strains MC3 and MA5 and was then subjected to an anaerobic digestion process, which increased the yield of methane by 140.32% and 158.68%, respectively, over that from nonpretreated microalgae. Therefore, a “whole-cell” cellulolytic pretreatment can increase the performance and efficiency of biogas production. PMID:24795376

  17. Enhancing the cellulose-degrading activity of cellulolytic bacteria CTL-6 (Clostridium thermocellum) by co-culture with non-cellulolytic bacteria W2-10 (Geobacillus sp.).

    Science.gov (United States)

    Lü, Yucai; Li, Ning; Yuan, Xufeng; Hua, Binbin; Wang, Jungang; Ishii, Masaharu; Igarashi, Yasuo; Cui, Zongjun

    2013-12-01

    The effect of a non-cellulolytic bacterium W2-10 (Geobacillus sp.) on the cellulose-degrading activity of a cellulolytic bacterium CTL-6 (Clostridium thermocellum) was determined using cellulose materials (paper and straw) in peptone cellulose solution (PCS) medium under aerobic conditions. The results indicated that in the co-culture, addition of W2-10 resulted in a balanced medium pH, and may provide the required anaerobic environment for CTL-6. Overall, addition of W2-10 was beneficial to CTL-6 growth in the adverse environment of the PCS medium. In co-culture with W2-10, the CTL-6 cellulose degradation efficiency of filter paper and alkaline-treated wheat straw significantly increased up to 72.45 and 37.79 %, respectively. The CMCase activity and biomass of CTL-6 also increased from 0.23 U ml(-1) and 45.1 μg ml(-1) (DNA content) up to 0.47 U ml(-1) and 112.2 μg ml(-1), respectively. In addition, co-culture resulted in accumulation of acetate and propionate up to 4.26 and 2.76 mg ml(-1). This was a respective increase of 2.58 and 4.45 times, in comparison to the monoculture with CTL-6.

  18. New Benzoxazine Secondary Metabolites from an Arctic Actinomycete

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    Kyuho Moon

    2014-04-01

    Full Text Available Two new secondary metabolites, arcticoside (1 and C-1027 chromophore-V (2, were isolated along with C-1027 chromophore-III and fijiolides A and B (3–5 from a culture of an Arctic marine actinomycete Streptomyces strain. The chemical structures of 1 and 2 were elucidated through NMR, mass, UV, and IR spectroscopy. The hexose moieties in 1 were determined to be d-glucose from a combination of acid hydrolysis, derivatization, and gas chromatographic analyses. Arcticoside (1 and C-1027 chromophore-V (2, which have a benzoxazine ring, inhibited Candida albicans isocitrate lyase. Chromophore-V (2 exhibited significant cytotoxicity against breast carcinoma MDA-MB231 cells and colorectal carcinoma cells (line HCT-116, with IC50 values of 0.9 and 2.7 μM, respectively.

  19. New benzoxazine secondary metabolites from an arctic actinomycete.

    Science.gov (United States)

    Moon, Kyuho; Ahn, Chan-Hong; Shin, Yoonho; Won, Tae Hyung; Ko, Keebeom; Lee, Sang Kook; Oh, Ki-Bong; Shin, Jongheon; Nam, Seung-Il; Oh, Dong-Chan

    2014-04-30

    Two new secondary metabolites, arcticoside (1) and C-1027 chromophore-V (2), were isolated along with C-1027 chromophore-III and fijiolides A and B (3-5) from a culture of an Arctic marine actinomycete Streptomyces strain. The chemical structures of 1 and 2 were elucidated through NMR, mass, UV, and IR spectroscopy. The hexose moieties in 1 were determined to be d-glucose from a combination of acid hydrolysis, derivatization, and gas chromatographic analyses. Arcticoside (1) and C-1027 chromophore-V (2), which have a benzoxazine ring, inhibited Candida albicans isocitrate lyase. Chromophore-V (2) exhibited significant cytotoxicity against breast carcinoma MDA-MB231 cells and colorectal carcinoma cells (line HCT-116), with IC₅₀ values of 0.9 and 2.7 μM, respectively.

  20. Isolation and identification of actinomycetes from a compost-amended soil with potential as biocontrol agents.

    Science.gov (United States)

    Cuesta, Gonzalo; García-de-la-Fuente, Rosana; Abad, Manuel; Fornes, Fernando

    2012-03-01

    The search for new biocontrol strategies to inhibit the growth of phytopathogenic microorganisms has become widely widespread due to environmental concerns. Among actinomycetes, Streptomyces species have been extensively studied since they have been recognized as important sources of antibiotics. Actinomycete strains were isolated from a calcareous soil, 2 two-phase olive mill waste ('alperujo') composts, and the compost-amended soil by using selective media, and they were then co-cultured with 5 phytopathogenic fungi and 1 bacterium to perform an in vitro antagonism assay. Forty-nine actinomycete strains were isolated, 12 of them showing a great antagonistic activity towards the phytopathogenic microorganisms tested. Isolated strains were identified by 16S rDNA sequence analysis and phenotypic procedures. Eleven isolates concerned the genus Streptomyces and 1 actinomycete with chitinolytic activity belonged to the genus Lechevalieria.

  1. Biodiversity, Anti-Trypanosomal Activity Screening, and Metabolomic Profiling of Actinomycetes Isolated from Mediterranean Sponges.

    Science.gov (United States)

    Cheng, Cheng; MacIntyre, Lynsey; Abdelmohsen, Usama Ramadan; Horn, Hannes; Polymenakou, Paraskevi N; Edrada-Ebel, RuAngelie; Hentschel, Ute

    2015-01-01

    Marine sponge-associated actinomycetes are considered as promising sources for the discovery of novel biologically active compounds. In the present study, a total of 64 actinomycetes were isolated from 12 different marine sponge species that had been collected offshore the islands of Milos and Crete, Greece, eastern Mediterranean. The isolates were affiliated to 23 genera representing 8 different suborders based on nearly full length 16S rRNA gene sequencing. Four putatively novel species belonging to genera Geodermatophilus, Microlunatus, Rhodococcus and Actinomycetospora were identified based on a 16S rRNA gene sequence similarity of actinomycete isolates showed bioactivities against Trypanosma brucei brucei TC221 with half maximal inhibitory concentration (IC50) values actinomycetes.

  2. Marine actinomycetes as an emerging resource for the drug development pipelines.

    Science.gov (United States)

    Zotchev, Sergey B

    2012-04-30

    Many representatives of the order Actinomycetales are prolific producers of thousands of biologically active secondary metabolites. Actinomycetes from terrestrial sources have been studied and screened since the 1950s, yielding many important anti-infective and anti-cancer drugs. However, frequent re-discovery of the same compounds in terrestrial actinomycetes have made them less attractive for screening programs in the recent years. At the same time, actinomycetes isolated from the marine environment currently receive considerable attention due to the structural diversity and unique biological activities of their secondary metabolites. This review highlights achievements and challenges in the isolation of marine actinomycetes, some examples of bioactive metabolites identified by conventional screening, and presents new developments in the field of genome mining and heterologous expression of biosynthetic gene clusters leading to the discovery of novel compounds. Copyright © 2011 Elsevier B.V. All rights reserved.

  3. Isolation strategies of marine-derived actinomycetes from sponge and sediment samples.

    Science.gov (United States)

    Hameş-Kocabaş, E Esin; Uzel, Ataç

    2012-03-01

    During the last two decades, discoveries of new members of actinomycetes and novel metabolites from marine environments have drawn attention to such environments, such as sediment and sponge. For the successful isolation of actinomycetes from marine environments, many factors including the use of enrichment and pre-treatment techniques, and the selection of growth media and antibiotic supplements should be taken into account. High-throughput cultivation is an innovative technique that mimics nature, eliminates undesired, fast-growing bacteria and creates suitable conditions for rare, slow-growing actinomycetes. This review comprehensively evaluates the traditional and innovative techniques and strategies used for the isolation of actinomycetes from marine sponge and sediment samples. Copyright © 2012 Elsevier B.V. All rights reserved.

  4. Mutational analysis of primary alcohol metabolism in the methylotrophic actinomycete Amycolatopsis methanolica

    NARCIS (Netherlands)

    Hektor, Harm J.; Dijkhuizen, Lubbert

    1996-01-01

    Mutants of the methylotrophic actinomycete Amycolatopsis methanolica unable to grow on methanol as carbon source were isolated and characterized. Mutants specifically affected in methanol utilization were deficient in formaldehyde assimilation. Mutants blocked in the first step of primary alcohol ox

  5. Antimicrobial activity of some actinomycetes from Western Ghats of Tamil Nadu, India

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    Pathalam Ganesan

    2017-06-01

    Conclusions: The present work revealed that, among 106 actinomycetes screened, Streptomyces rimosus (FMS-20 (Accession No-KT827106 showed promising antimicrobial activity against all the tested human microbial pathogens.

  6. Exploring plant growth-promotion actinomycetes from vermicompost and rhizosphere soil for yield enhancement in chickpea.

    Science.gov (United States)

    Sreevidya, M; Gopalakrishnan, S; Kudapa, H; Varshney, R K

    2016-01-01

    The main objective of the present study was to isolate and characterize actinomycetes for their plant growth-promotion in chickpea. A total of 89 actinomycetes were screened for their antagonism against fungal pathogens of chickpea by dual culture and metabolite production assays. Four most promising actinomycetes were evaluated for their physiological and plant growth-promotion properties under in vitro and in vivo conditions. All the isolates exhibited good growth at temperatures from 20°C to 40°C, pH range of 7-11 and NaCl concentrations up to 8%. These were also found highly tolerant to Bavistin, slightly tolerant to Thiram and Captan (except VAI-7 and VAI-40) but susceptible to Benlate and Ridomil at field application levels and were found to produce siderophore, cellulase, lipase, protease, chitinase (except VAI-40), hydrocyanic acid (except VAI-7 and VAI-40), indole acetic acid and β-1,3-glucanase. When the four actinomycetes were evaluated for their plant growth-promotion properties under field conditions on chickpea, all exhibited increase in nodule number, shoot weight and yield. The actinomycetes treated plots enhanced total N, available P and organic C over the un-inoculated control. The scanning electron microscope studies exhibited extensive colonization by actinomycetes on the root surface of chickpea. The expression profiles for indole acetic acid, siderophore and β-1,3-glucanase genes exhibited up-regulation for all three traits and in all four isolates. The actinomycetes were identified as Streptomyces but different species in the 16S rDNA analysis. It was concluded that the selected actinomycetes have good plant growth-promotion and biocontrol potentials on chickpea.

  7. Isolation and Characterization of Actinomycete Antagonists of a Fungal Root Pathogen †

    Science.gov (United States)

    Crawford, Don L.; Lynch, James M.; Whipps, John M.; Ousley, Margaret A.

    1993-01-01

    By use of selective media, 267 actinomycete strains were isolated from four rhizosphere-associated and four non-rhizosphere-associated British soils. Organic media with low nutrient concentrations were found to be best for isolating diverse actinomycetes while avoiding contamination and overgrowth of isolation media by eubacteria and fungi. While all isolates grew well at pHs 6.5 to 8.0, a few were unable to grow at pH 6.0 and a significant number failed to grow at pH 5.5. Eighty-two selected isolates were screened for in vitro antagonism towards Pythium ultimum by use of a Difco cornmeal agar assay procedure. Five isolates were very strong antagonists of the fungus, four were strong antagonists, and ten others were weakly antagonistic. The remaining isolates showed no antagonism by this assay. Additional studies showed that several of the P. ultimum antagonists also strongly inhibited growth of other root-pathogenic fungi. Twelve isolates showing antifungal activity in the in vitro assay were also tested for their effects on the germination and short-term growth of lettuce plants in glasshouse pot studies in the absence of pathogens. None of the actinomycetes prevented seed germination, although half of the isolates retarded seed germination and outgrowth of the plants by 1 to 3 days. During 18-day growth experiments, biomass yields of some actinomycete-inoculated plants were reduced in comparison with untreated control plants, although all plants appeared healthy and well rooted. None of the actinomycetes significantly enhanced plant growth over these short-term experiments. For some, but not all, actinomycetes, some correlations between delayed seed germination and reduced 18-day plant biomass yields were seen. For others, plant biomass yields were not reduced despite an actinomycete-associated delay in seed germination and plant outgrowth. Preliminary glasshouse experiments indicated that some of the actinomycetes protect germinating lettuce seeds against

  8. Exploring plant growth-promotion actinomycetes from vermicompost and rhizosphere soil for yield enhancement in chickpea

    Directory of Open Access Journals (Sweden)

    M. Sreevidya

    2016-03-01

    Full Text Available Abstract The main objective of the present study was to isolate and characterize actinomycetes for their plant growth-promotion in chickpea. A total of 89 actinomycetes were screened for their antagonism against fungal pathogens of chickpea by dual culture and metabolite production assays. Four most promising actinomycetes were evaluated for their physiological and plant growth-promotion properties under in vitro and in vivo conditions. All the isolates exhibited good growth at temperatures from 20 °C to 40 °C, pH range of 7–11 and NaCl concentrations up to 8%. These were also found highly tolerant to Bavistin, slightly tolerant to Thiram and Captan (except VAI-7 and VAI-40 but susceptible to Benlate and Ridomil at field application levels and were found to produce siderophore, cellulase, lipase, protease, chitinase (except VAI-40, hydrocyanic acid (except VAI-7 and VAI-40, indole acetic acid and β-1,3-glucanase. When the four actinomycetes were evaluated for their plant growth-promotion properties under field conditions on chickpea, all exhibited increase in nodule number, shoot weight and yield. The actinomycetes treated plots enhanced total N, available P and organic C over the un-inoculated control. The scanning electron microscope studies exhibited extensive colonization by actinomycetes on the root surface of chickpea. The expression profiles for indole acetic acid, siderophore and β-1,3-glucanase genes exhibited up-regulation for all three traits and in all four isolates. The actinomycetes were identified as Streptomyces but different species in the 16S rDNA analysis. It was concluded that the selected actinomycetes have good plant growth-promotion and biocontrol potentials on chickpea.

  9. Stimulation of 2-methylisoborneol (MIB) production by actinomycetes after cyclic chlorination in drinking water distribution systems.

    Science.gov (United States)

    Abbaszadegan, Morteza; Yi, Min; Alum, Absar

    2015-01-01

    The impact of fluctuation in chlorine residual on actinomycetes and the production of 2-methylisoborneol (MIB) were studied in cast-iron and PVC model distribution systems. Actinomycetes were spiked in each system and continued operation for a 12-day non-chlorine experiment, resulting in no changes in actinomycetes and MIB concentrations. Three cyclic chlorination events were performed and chlorine residuals were maintained as follows: 1.0 mg L(-1) for 24 h, 0 mg L(-1) for 48 h, 0.5 mg L(-1) for 48 h, 0 mg L(-1) for 48 h and 2 mg L(-1) for 24 h. After each chlorination event, 2 -3 log decrease in actinomycetes was noted in both systems. However, within 48 h at 0 mg L(-1) chlorine, the actinomycetes recovered to the pre-chlorination levels. On the contrary, MIB concentration in both systems remained un-impacted after the first cycle and increased by fourfold ( 20 mg L(-1)) after the second cycle, which lasted through the third cycle despite the fact that actinomycetes numbers fluctuated 2-3 logs during this time period. For obtaining biofilm samples from field, water meters were collected from municipality drinking water distribution systems located in central Arizona. The actinomycetes concentration in asbestos cement pipe and cast iron pipe averaged 3.1 × 10(3) and 1.9 × 10(4) CFU cm(-2), respectively. The study shows that production of MIB is associated with changes in chlorine residual in the systems. This is the first report of cyclic chlorine shock as a stimulus for MIB production by actinomycetes in drinking water distribution system's ecology.

  10. Targeted search for actinomycetes from near-shore and deep-sea marine sediments

    OpenAIRE

    Prieto-Davó, Alejandra; Villarreal-Gómez, Luis Jesús; Forschner-Dancause, Stephanie; Bull, Alan T.; Stach, James E. M.; David C. Smith; Rowley, Dave C.; Jensen, Paul R.

    2013-01-01

    Sediment samples collected off the coast of San Diego were analyzed for actinomycete diversity using culture independent techniques. Eight new operational taxonomic units (OTUs) in the Streptomycetaceae were identified as well as new diversity within previously cultured marine OTUs. Sequences belonging to the marine actinomycete genus Salinispora were also detected, despite the fact that this genus has only been reported from more tropical environments. Independent analyses of marine sediment...

  11. Identification of the Entner-Doudoroff pathway in an antibiotic-producing actinomycete species

    DEFF Research Database (Denmark)

    Gunnarsson, Nina; Mortensen, Uffe Hasbro; Sosio, M.

    2004-01-01

    the primary metabolic pathways of the poorly characterized antibiotic-producing actinomycete Nonomuraea sp. ATCC 39727. Surprisingly, it was found that Nonomuraea sp. ATCC 39272 predominantly metabolizes glucose via the Entner-Doudoroff (ED) pathway. This represents the first time that the ED pathway has been...... recognized as the main catabolic pathway in an actinomycete. The Nonomuraea genes encoding the key enzymes of the ED pathway were subsequently identified, sequenced and functionally described....

  12. Actinomycetes from Western Ghats of Tamil Nadu with its antimicrobial properties

    Institute of Scientific and Technical Information of China (English)

    Valan Arasu M; Ignacimuthu S; Agastian P

    2012-01-01

    Objective: To isolate the actinomycetes from Western Ghats of Tamil Nadu with its antimicrobial properties. Methods: Starch casein agar medium supplemented with actidione and nalidixic acid was used to isolate actinomycetes from Western Ghates region of Kanyakumari, Thirunelveli, Dindigul and Nilgiri districts. Modified nutrient medium was used as the base for screening actinomycetes against pathogenic Gram positive, Gram negative and filamentous fungi. Results:Among 367 actinomycetes; 17.71% showed activity against both bacteria and fungi. The highest antibacterial activity was observed against B. subtilis, 140 isolates (38.1%), S. aureus 128 (34.9%);S. epidermidis 123 (33.5%); P. aeruginosa 105 (28.6%); K. pneumoniae 88 (24%); Xanthomonas sp 62 (16.9%). Less number of actinomycetes showed activity against Erwinia, S. typhi, V. fischeri andP. vulgaris. Hundred and three isolates showed activity against B. cinerea and A. niger. Twenty five isolates revealed activity against T. simii. Conclusions: Present investigation concludes that Western Ghats region of Tamil Nadu is the potential place for actinomycetes diversity. Further studying about these medically important strains from this region can be useful in identification of valuable bio-molecules.

  13. Enrichment Method for the Isolation of Bioactive Actinomycetes From Mangrove Sediments of Andaman Islands, India

    Directory of Open Access Journals (Sweden)

    Baskaran, R.

    2011-01-01

    Full Text Available Various pre-treatment methods and three different media were employed for the isolation of bioactive actinomycetes from mangrove sediments of Andaman and Nicobar Islands, India. Sediments from four different sites of mangrove forest were collected and pre-treated by dry heat method, and the media were supplemented with cycloheximide 80 µg/mL and nalidixic acid 75 µg/mL. The mean actinomycetes population density in sediment samples were recorded as 22 CFU-10^-6/gm in KUA medium followed by 12 CFU-10^-6/gm in AIA medium and 8 CFU-10^-6/gm in SCA medium. A total of 42 actinomycetes were isolated, and all the isolates were evaluated for their antibacterial activity against pathogenic bacteria on two different media. Among 42 isolates tested, 22 species were found to be antibacterial metabolite producer against test bacteria namely, Staphylococcus aureus, Bacillus subtilis, Salmonella typhi and Klebsiella pneumoniae. Particularly, the actinomycete strains such as A101, A102, A107, A116, A121, A125, A130, F101, F102, F104, F106, De101 and De102 significantly inhibited the growth of all bacteria which were tested. Of these strains, A107 was identified as Streptomyces spp. This strain had the maximum activity against all used pathogens on both medium. Hence, the isolation, characterization and studies of secondary metabolites of actinomycetes from mangrove sediments in Andaman and Nicobar Island could be a pathway for discovery of antibiotics from marine actinomycetes.

  14. Isolation and characterization of medically important aerobic actinomycetes in soil of iran (2006 - 2007).

    Science.gov (United States)

    Aghamirian, Mohammad Reza; Ghiasian, Seyed Amir

    2009-01-01

    The aerobic actinomycetes are a large group of soil-inhabiting bacteria that occur worldwide. Some of them are the main cause of two important diseases, nocardiosis and actinomycetoma. To identify the prevalence and geographic distribution of aerobic actinomycetes in soil of Qazvin province, a study was carried out during 2006-2007. In this study, the incidence and diversity of medically important aerobic actinomycetes was determined in 300 soil samples of different parts of Qazvin. The suspensions of superficial soil samples were prepared by adding of normal saline, streptomycin and chloramphenicol and the supernatants were cultured on brain-heart infusion agar and Sabouraud's dextrose agar contain cycloheximide. The isolated microorganisms were examined by Gram and acid-fast stains and were identified biochemically and morphologically. Of 96 aerobic actinomycetes isolates identified, Actinomadura madurae and Streptomyces somaliensis were the most frequently isolated species each representing 19.8% of isolates, followed by Nocardia asteroides (15.6%), N. otitidiscaviarum (9.4%), N. brasiliensis (7.3%), A. peletieri, S. griseus, and Nocardia spp. (each 5.2%), and N. transvalensis, Nocardiopsis dassonvillei, Actinomadura spp. and Streptomyces spp. (each 3.1%). To the best of our knowledge, this is the first report on epidemiological investigation of medically important aerobic actinomycetes in soil samples from Iran. In recent years, mycetoma and nocardiosis have been increasingly reported in Iran. The results showed that medically important actinomycetes occur in the environment of Iran and soil could be potential source of actinomycotic infections.

  15. Actinomycetal complex of light sierozem on the Kopet-Dag piedmont plain

    Science.gov (United States)

    Zenova, G. M.; Zvyagintsev, D. G.; Manucharova, N. A.; Stepanova, O. A.; Chernov, I. Yu.

    2016-10-01

    The population density of actinomycetes in the samples of light sierozem from the Kopet Dag piedmont plain (75 km from Ashkhabad, Turkmenistan) reaches hundreds of thousand CFU/g soil. The actinomycetal complex is represented by two genera: Streptomyces and Micromonospora. Representatives of the Streptomyces genus predominate and comprise 73 to 87% of the actinomycetal complex. In one sample, representatives of the Micromonospora genus predominated in the complex (75%). The Streptomyces genus in the studied soil samples is represented by the species from several sections and series: the species of section Helvolo-Flavus series Helvolus represent the dominant component of the streptomycetal complex; their portion is up to 77% of all isolated actinomycetes. The species of other sections and series are much less abundant. Thus, the percentage of the Cinereus Achromogenes section in the actinomycetal complex does not exceed 28%; representatives of the Albus section Albus series, Roseus section Lavendulae-Roseus series, and Imperfectus section belong to rare species; they have been isolated not from all the studied samples of light sierozem, and their portion does not exceed 10% of the actinomycetal complex.

  16. Diversity of Nonribosomal Peptide Synthetase Genes in the AnticancerProducing Actinomycetes Isolated from Marine Sediment in Indonesia

    OpenAIRE

    Camelia Herdini; Shinta Hartanto; Sofia Mubarika; Bambang Hariwiyanto; Nastiti Wijayanti; Akira Hosoyama; Atsushi Yamazoe; Hideaki Nojiri; Jaka Widada

    2016-01-01

    Marine actinomycetes is a group of bacteria that is highly potential in producing novel bioactive compound. It has unique characteristics and is different from other terrestrial ones. Extreme environmental condition is suspected to lead marine actinomycetes produce different types of bioactive compound found previously. The aim of this study was to explore the presence and diversity of NRPS genes in 14 anticancer-producing actinomycetes isolated from marine sediment in Indonesia. ...

  17. Draft Genome Sequences of Three Cellulolytic Bacillus licheniformis Strains Isolated from Imperial Geyser, Amphitheater Springs, and Whiterock Springs inside Yellowstone National Park

    Science.gov (United States)

    O' Hair, Joshua A.; Li, Hui; Thapa, Santosh; Scholz, Matthew

    2017-01-01

    ABSTRACT Novel cellulolytic microorganisms are becoming more important for rapidly growing biofuel industries. This paper reports the draft genome sequences of Bacillus licheniformis strains YNP2-TSU, YNP3-TSU, and YNP5-TSU. These cellulolytic isolates were collected from several hydrothermal features inside Yellowstone National Park. PMID:28360181

  18. Screening and characterization of protease producing actinomycetes from marine saltern.

    Science.gov (United States)

    Suthindhiran, Krish; Jayasri, Mangalam Achuthananda; Dipali, Dipa; Prasar, Apurva

    2014-10-01

    In the course of systematic screening program for bioactive actinomycetes, an alkaline protease producing halophilic strain Actinopolyspora sp. VITSDK2 was isolated from marine saltern, Southern India. The strain was identified as Actinopolyspora based on its phenotypic and phylogenetic characters. The protease was partially purified using ammonium sulfate precipitation and subsequently by DEAE cellulose column chromatography. The enzyme was further purified using HPLC and the molecular weight was found to be 22 kDa as determined by SDS-PAGE analysis. The purified protease exhibited pH stability in a wide range of 4-12 with optimum at 10.0. The enzyme was found to be stable between 25 and 80 °C and displayed a maximum activity at 60 °C. The enzyme activity was increased marginally in presence of Mn(2+) , Mg(2+) , and Ca(2+) and decreased in presence of Cu(2+) . PMSF and DFP completely inhibited the activity suggesting it belongs to serine protease. Further, the proteolytic activity was abolished in presence of N-tosyl-L-lysine chloromethyl ketone suggesting this might be chymotrypsin-like serine protease. The protease was 96% active when kept for 10 days at room temperature. The results indicate that the enzyme belong to chymotrypsin-like serine protease exhibiting both pH and thermostability, which can be used for various applications in industries. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Rhodococcus equi: the many facets of a pathogenic actinomycete.

    Science.gov (United States)

    Vázquez-Boland, José A; Giguère, Steeve; Hapeshi, Alexia; MacArthur, Iain; Anastasi, Elisa; Valero-Rello, Ana

    2013-11-29

    Rhodococcus equi is a soil-dwelling pathogenic actinomycete that causes pulmonary and extrapulmonary pyogranulomatous infections in a variety of animal species and people. Young foals are particularly susceptible and develop a life-threatening pneumonic disease that is endemic at many horse-breeding farms worldwide. R. equi is a facultative intracellular parasite of macrophages that replicates within a modified phagocytic vacuole. Its pathogenicity depends on a virulence plasmid that promotes intracellular survival by preventing phagosome-lysosome fusion. Species-specific tropism of R. equi for horses, pigs and cattle appears to be determined by host-adapted virulence plasmid types. Molecular epidemiological studies of these plasmids suggest that human R. equi infection is zoonotic. Analysis of the recently determined R. equi genome sequence has identified additional virulence determinants on the bacterial chromosome. This review summarizes our current understanding of the clinical aspects, biology, pathogenesis and immunity of this fascinating microbe with plasmid-governed infectivity. Copyright © 2013 Elsevier B.V. All rights reserved.

  20. Cellulolytic enzymes, nucleic acids encoding them and methods for making and using them

    Science.gov (United States)

    Gray, Kevin A [San Diego, CA; Zhao, Lishan [Emeryville, CA; Cayouette, Michelle H [San Diego, CA

    2012-01-24

    The invention provides polypeptides having any cellulolytic activity, e.g., a cellulase activity, a endoglucanase, a cellobiohydrolase, a beta-glucosidase, a xylanase, a mannanse, a .beta.-xylosidase, an arabinofuranosidase, and/or an oligomerase activity, polynucleotides encoding these polypeptides, and methods of making and using these polynucleotides and polypeptides. In one aspect, the invention is directed to polypeptides having any cellulolytic activity, e.g., a cellulase activity, e.g., endoglucanase, cellobiohydrolase, beta-glucosidase, xylanase, mannanse, .beta.-xylosidase, arabinofuranosidase, and/or oligomerase activity, including thermostable and thermotolerant activity, and polynucleotides encoding these enzymes, and making and using these polynucleotides and polypeptides. In one aspect, the invention provides polypeptides having an oligomerase activity, e.g., enzymes that convert recalcitrant soluble oligomers to fermentable sugars in the saccharification of biomass. The polypeptides of the invention can be used in a variety of pharmaceutical, agricultural, food and feed processing and industrial contexts. The invention also provides compositions or products of manufacture comprising mixtures of enzymes comprising at least one enzyme of this invention.

  1. A potential source for cellulolytic enzyme discovery and environmental aspects revealed through metagenomics of Brazilian mangroves.

    Science.gov (United States)

    Thompson, Claudia Elizabeth; Beys-da-Silva, Walter Orlando; Santi, Lucélia; Berger, Markus; Vainstein, Marilene Henning; Guima Rães, Jorge Almeida; Vasconcelos, Ana Tereza Ribeiro

    2013-10-26

    The mangroves are among the most productive and biologically important environments. The possible presence of cellulolytic enzymes and microorganisms useful for biomass degradation as well as taxonomic and functional aspects of two Brazilian mangroves were evaluated using cultivation and metagenomic approaches. From a total of 296 microorganisms with visual differences in colony morphology and growth (including bacteria, yeast and filamentous fungus), 179 (60.5%) and 117 (39.5%) were isolated from the Rio de Janeiro (RJ) and Bahia (BA) samples, respectively. RJ metagenome showed the higher number of microbial isolates, which is consistent with its most conserved state and higher diversity. The metagenomic sequencing data showed similar predominant bacterial phyla in the BA and RJ mangroves with an abundance of Proteobacteria (57.8% and 44.6%), Firmicutes (11% and 12.3%) and Actinobacteria (8.4% and 7.5%). A higher number of enzymes involved in the degradation of polycyclic aromatic compounds were found in the BA mangrove. Specific sequences involved in the cellulolytic degradation, belonging to cellulases, hemicellulases, carbohydrate binding domains, dockerins and cohesins were identified, and it was possible to isolate cultivable fungi and bacteria related to biomass decomposition and with potential applications for the production of biofuels. These results showed that the mangroves possess all fundamental molecular tools required for building the cellulosome, which is required for the efficient degradation of cellulose material and sugar release.

  2. In vitro Cellulose Rich Organic Material Degradation by Cellulolytic Streptomyces albospinus (MTCC 8768

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    Pinky Prasad

    2012-09-01

    Full Text Available Aims: Cellulosic biomass is the only foreseeable sustainable source of fuels and is also one of the dominating waste materials in nature resulting from human activities. Keeping in view the environmental problems like disposal of large volumes of cellulosic wastes and shortage of fossil fuel in the world, the main aim of the present investigation was to characterize and study the cellulolytic activity of Streptomyces albospinus (MTCC 8768, isolated from municipal wastes, on natural cellulosic substrates viz. straw powder, wood powder and finely grated vegetable peels.Methodology and Result: Stanier’s Basal broth with 100 mg of each of the substrates was inoculated separately with S. albospinus (MTCC No. 8768 and incubated at 37 °C for 8 days. The cellulosic substrates were re-weighed at an interval of 2 days and the difference between the initial weight and the final weight gave the amount of substratesdegraded by the isolate. It was observed that maximum degradation was observed in the grated vegetable peels (64 mg followed by straw powder (38 mg and wood powder (28 mg over a period of 8 days.Conclusion, significance and impact of study: By the selection of efficient cellulolytic microorganisms and cost-effective operational techniques, the production of useful end products from the biodegradation of the low cost enormous stock of cellulose in nature can be very beneficial.

  3. Diarrhea-associated pathogens, lactobacilli and cellulolytic bacteria in equine feces: responses to antibiotic challenge.

    Science.gov (United States)

    Harlow, Brittany E; Lawrence, Laurie M; Flythe, Michael D

    2013-09-27

    Antibiotics are important to equine medicine, but antibiotic-associated diarrhea (AAD) can lead to poor performance and even mortality. AAD is attributed to disruption of the hindgut microbiota, which permits proliferation of pathogenic microbes. The goal of this study was to evaluate the effects of common antibiotics on cellulolytic bacteria, lactobacilli, and AAD-associated pathogens in the feces of healthy horses. Fifteen horses were assigned to three treatment groups (blocked by age and sex): control (no antibiotics), trimethoprim-sulfadiazine (PO), or ceftiofur (IM). Fecal samples (n=8 per horse) were taken during dietary adaptation (3 weeks), antibiotic challenge (1 week), and withdrawal (1 week). Bacteria were enumerated by serial dilution and viable count. Cellulolytic bacteria decreased by >99% during administration of either antibiotic (Pantibiotic challenge period (PAntibiotic challenged horses also shed more salmonella than control horses (PAntibiotics had no effect on the number of Clostridium perfringens isolates. There was no detectable Clostridium difficile during adaptation or in any control horse. C. difficile increased (Pantibiotics, and were still detectable 1 week after withdrawal. These results indicate that antibiotics can disrupt the normal gastrointestinal microbiota and allow proliferation of Salmonella spp. and C. difficile.

  4. Clostridium herbivorans sp. nov., a cellulolytic anaerobe from the pig intestine.

    Science.gov (United States)

    Varel, V H; Tanner, R S; Woese, C R

    1995-07-01

    A new cellulolytic anaerobic clostridium was isolated from the intestinal tract of pigs. The single isolate was a gram-positive, motile rod, formed terminal to subterminal swollen sporangia, and required a fermentable carbohydrate for growth. Cellulose, cellobiose, maltose, starch, and glycogen supported growth, but glucose and fructose did not. The major end products from the fermentation of cellobiose were butyrate and formate; minor amounts of hydrogen and ethanol were also formed. Ruminal fluid (15%) or yeast extract (1%) was required for good growth. The optimum temperature for growth was 39 to 42 degrees C, and the optimum pH was 6.8 to 7.2. Cell lysis occurred rapidly once stationary growth was reached. A 16S rRNA sequence analysis showed that the strain was related to a group of gram-positive anaerobes that includes Clostridium oroticum and the cellulolytic species Clostridium polysaccharolyticum and Clostridium populeti. The DNA base composition of the isolate is 38 mol% G + C. We propose the name Clostridium herbivorans for this organism; strain 54408 (= ATCC 49925) is the type strain.

  5. Industrial waste based compost as a source of novel cellulolytic strains and enzymes.

    Science.gov (United States)

    Amore, Antonella; Pepe, Olimpia; Ventorino, Valeria; Birolo, Leila; Giangrande, Chiara; Faraco, Vincenza

    2013-02-01

    Ninety bacteria isolated from raw composting materials were screened for their cellulolytic activity on solid medium containing carboxymethylcellulose. The bacteria producing the highest cellulolytic activity levels were identified by 16S rRNA sequencing as Bacillus licheniformis strain 1, Bacillus subtilis subsp. subtilis strain B7B, Bacillus subtilis subsp. spizizenii strain 6, and Bacillus amyloliquefaciens strain B31C. Cellulase activity production by the most productive strain B. amyloliquefaciens B31C was optimized in liquid culture varying the carbon source. Comparison of growth curves of B. amyloliquefaciens B31C at temperatures from 28 to 47 °C indicated its thermotolerant nature. Moreover, analysis of time courses of cellulase activity production in this thermal range showed that increase of temperature from 28 to 37 °C causes an increase of cellulase activity levels. Investigating the enzymes responsible for cellulase activity produced by B. amyloliquefaciens B31C by proteomic analyses, an endoglucanase was identified. It was shown that the purified enzyme catalyzes carboxymethylcellulose's hydrolysis following Michaelis-Menten kinetics with a K(M) of 9.95 mg ml(-1) and a v(max) of 284 μM min(-1) . It shows a retention of 90% of its activity for at least 144 h of incubation at 40 °C and exhibits a range of optimum temperatures from 50 to 70 °C.

  6. Biodegradation of Palm Kernel Cake by Cellulolytic and Hemicellulolytic Bacterial Cultures through Solid State Fermentation

    Directory of Open Access Journals (Sweden)

    Mohamed Idris Alshelmani

    2014-01-01

    Full Text Available Four cellulolytic and hemicellulolytic bacterial cultures were purchased from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Culture (DSMZ and the American Type Culture Collection (ATCC. Two experiments were conducted; the objective of the first experiment was to determine the optimum time period required for solid state fermentation (SSF of palm kernel cake (PKC, whereas the objective of the second experiment was to investigate the effect of combinations of these cellulolytic and hemicellulolytic bacteria on the nutritive quality of the PKC. In the first experiment, the SSF was lasted for 12 days with inoculum size of 10% (v/w on different PKC to moisture ratios. In the second experiment, fifteen combinations were created among the four microbes with one untreated PKC as a control. The SSF lasted for 9 days, and the samples were autoclaved, dried, and analyzed for proximate analysis. Results showed that bacterial cultures produced high enzymes activities at the 4th day of SSF, whereas their abilities to produce enzymes tended to be decreased to reach zero at the 8th day of SSF. Findings in the second experiment showed that hemicellulose and cellulose was significantly P<0.05 decreased, whereas the amount of reducing sugars were significantly P<0.05 increased in the fermented PKC (FPKC compared with untreated PKC.

  7. Actinomycetes from Sediments in the Trondheim Fjord, Norway: Diversity and Biological Activity

    Directory of Open Access Journals (Sweden)

    Sergey B. Zotchev

    2008-02-01

    Full Text Available The marine environment represents a largely untapped source for isolation of new microorganisms with potential to produce biologically active secondary metabolites. Among such microorganisms, Gram-positive actinomycete bacteria are of special interest, since they are known to produce chemically diverse compounds with a wide range of biological activities. We have set out to isolate and characterize actinomycete bacteria from the sediments in one of the largest Norwegian fjords, the Trondheim fjord, with respect to diversity and antibiotic-producing potential. Approximately 3,200 actinomycete bacteria were isolated using four different agar media from the sediment samples collected at different locations and depths (4.5 to 450 m. Grouping of the isolates first according to the morphology followed by characterization of isolates chosen as group representatives by molecular taxonomy revealed that Micromonospora was the dominating actinomycete genus isolated from the sediments. The deep water sediments contained a higher relative amount of Micromonospora compared to the shallow water samples. Nine percent of the isolates clearly required sea water for normal growth, suggesting that these strains represent obligate marine organisms. Extensive screening of the extracts from all collected isolates for antibacterial and antifungal activities revealed strong antibiotic-producing potential among them. The latter implies that actinomycetes from marine sediments in Norwegian fjords can be potential sources for the discovery of novel anti-infective agents.

  8. Diversity and bioprospecting of culturable actinomycetes from marine sediment of the Yellow Sea, China.

    Science.gov (United States)

    Xiong, Zhi-Qiang; Liu, Qiao-Xia; Pan, Zhao-Long; Zhao, Na; Feng, Zhi-Xiang; Wang, Yong

    2015-03-01

    Marine actinomycetes are a potential source of a wide variety of bioactive natural products. In this work, seven pretreatments, three selective isolation media, and five artificial seawater concentrations were used to isolate actinomycetes from the sediments collected from Yellow Sea, China. Statistical analysis showed that only the isolation medium strongly affected the total and bioactive numbers of actinomycete isolates. A total of 613 actinobacterial strains were isolated and screened for antimicrobial activities; 154 isolates showed activity against at least one of nine test drug-resistant microorganisms. Eighty-nine representatives with strong antimicrobial activity were identified phylogenetically based on 16S rRNA gene sequencing, which were assigned to five different actinomycete genera Streptomyces, Kocuria, Saccharomonospora, Micromonospora, and Nocardiopsis. Using PCR-based screening for six biosynthetic genes of secondary metabolites, all 45 isolates with acute activity have at least one biosynthetic gene, 28.8 % of which possess more than three biosynthetic genes. As a case, strain SMA-1 was selected for antimicrobial natural product discovery. Three diketopiperazine dimers including a new compound iso-naseseazine B (1) and two known compounds naseseazine B (2) and aspergilazine A (3) were isolated by bioassay-guided separation. These results suggested that actinomycetes from marine sediments are a potential resource of novel secondary metabolites and drugs.

  9. Actinomycetes from the South China Sea sponges: isolation, diversity and potential for aromatic polyketides discovery

    Directory of Open Access Journals (Sweden)

    Zhiyong eLi

    2015-10-01

    Full Text Available Marine sponges often harbor dense and diverse microbial communities including actinobacteria. To date no comprehensive investigation has been performed on the culturable diversity of the actinomycetes associated with South China Sea sponges. Structurally novel aromatic polyketides were recently discovered from marine sponge-derived Streptomyces and Saccharopolyspora strains, suggesting that sponge-associated actinomycetes can serve as a new source of aromatic polyketides. In this study, a total of 77 actinomycete strains were isolated from 15 South China Sea sponge species. Phylogenetic characterization of the isolates based on 16S rRNA gene sequencing supported their assignment to 12 families and 20 genera, among which three rare genera (Marihabitans, Polymorphospora and Streptomonospora were isolated from marine sponges for the first time. Subsequently, β-ketoacyl synthase (KSα gene was used as marker for evaluating the potential of the actinomycete strains to produce aromatic polyketides. As a result, KSα gene was detected in 35 isolates related to 7 genera (Kocuria, Micromonospora, Nocardia, Nocardiopsis, Saccharopolyspora, Salinispora and Streptomyces. Finally, ten strains were selected for small-scale fermentation, and one angucycline compound was detected from the culture extract of Streptomyces anulatus strain S71. This study advanced our knowledge of the sponge-associated actinomycetes regarding their diversity and potential in producing aromatic polyketides.

  10. Exploring the diversity and metabolic potential of actinomycetes from temperate marine sediments from Newfoundland, Canada.

    Science.gov (United States)

    Duncan, K R; Haltli, B; Gill, K A; Correa, H; Berrué, F; Kerr, R G

    2015-01-01

    Marine sediments from Newfoundland, Canada were explored for biotechnologically promising Actinobacteria using culture-independent and culture-dependent approaches. Culture-independent pyrosequencing analyses uncovered significant actinobacterial diversity (H'-2.45 to 3.76), although the taxonomic diversity of biotechnologically important actinomycetes could not be fully elucidated due to limited sampling depth. Assessment of culturable actinomycete diversity resulted in the isolation of 360 actinomycetes representing 59 operational taxonomic units, the majority of which (94 %) were Streptomyces. The biotechnological potential of actinomycetes from NL sediments was assessed by bioactivity and metabolomics-based screening of 32 representative isolates. Bioactivity was exhibited by 41 % of isolates, while 11 % exhibited unique chemical signatures in metabolomics screening. Chemical analysis of two isolates resulted in the isolation of the cytotoxic metabolite 1-isopentadecanoyl-3β-D-glucopyranosyl-X-glycerol from Actinoalloteichus sp. 2L868 and sungsanpin from Streptomyces sp. 8LB7. These results demonstrate the potential for the discovery of novel bioactive metabolites from actinomycetes isolated from Atlantic Canadian marine sediments.

  11. Isolation of actinomycetes from mangrove and estuarine sediments of Cochin and screening for antimicrobial activity

    Directory of Open Access Journals (Sweden)

    Emilda Rosmine

    2016-03-01

    Full Text Available Objective: To isolate and screen actinomycetes for antimicrobial activity from mangroves and estuarine soil samples of Cochin. Methods: In the present study, sediment samples collected from mangroves and various stations of Cochin estuary were pretreated and actinomycetes were isolated on different selective media. The isolates were screened for antibiotic activity by following disc diffusion assay (Kirby-Bauer method against human pathogens, fish pathogens and Gram-positive bacteria. The isolates were identified based on their morphology. Results: Only 2 actinomycete isolates (ER7 and ER10 of the 50 isolates screened had antimicrobial activities against one or more pathogens tested. ER7 isolate showed higher antimicrobial activity as compared to that of ER10 isolate. The maximum inhibition zone of crude extract from ER7 was 16.7 mm. The methanol extract of ER7 showed antimicrobial activity against all the pathogens tested with a maximum zone of 21.0 mm. The isolates with antimicrobial activity were found to belong to the genus Streptomyces. Conclusions: There is no significant report on bioactive actinomycetes from the present study areas. Potent antibiotics from the selected isolates could contribute to fight against several human and fish diseases. Further purification, structural elucidation and characterization are recommended to know the quality, novelty and commercial value of these antibiotics. Hence, the mangroves and estuary of Kochi show great promise for the discovery of bioactive actinomycetes.

  12. Actinomycetes from Red Sea sponges: sources for chemical and phylogenetic diversity.

    Science.gov (United States)

    Abdelmohsen, Usama Ramadan; Yang, Chen; Horn, Hannes; Hajjar, Dina; Ravasi, Timothy; Hentschel, Ute

    2014-05-12

    The diversity of actinomycetes associated with marine sponges collected off Fsar Reef (Saudi Arabia) was investigated in the present study. Forty-seven actinomycetes were cultivated and phylogenetically identified based on 16S rRNA gene sequencing and were assigned to 10 different actinomycete genera. Eight putatively novel species belonging to genera Kocuria, Mycobacterium, Nocardia, and Rhodococcus were identified based on sequence similarity values below 98.2% to other 16S rRNA gene sequences available in the NCBI database. PCR-based screening for biosynthetic genes including type I and type II polyketide synthases (PKS-I, PKS-II) as well as nonribosomal peptide synthetases (NRPS) showed that 20 actinomycete isolates encoded each at least one type of biosynthetic gene. The organic extracts of nine isolates displayed bioactivity against at least one of the test pathogens, which were Gram-positive and Gram-negative bacteria, fungi, human parasites, as well as in a West Nile Virus protease enzymatic assay. These results emphasize that marine sponges are a prolific resource for novel bioactive actinomycetes with potential for drug discovery.

  13. Actinomycetes from Red Sea Sponges: Sources for Chemical and Phylogenetic Diversity

    Directory of Open Access Journals (Sweden)

    Usama Ramadan Abdelmohsen

    2014-05-01

    Full Text Available The diversity of actinomycetes associated with marine sponges collected off Fsar Reef (Saudi Arabia was investigated in the present study. Forty-seven actinomycetes were cultivated and phylogenetically identified based on 16S rRNA gene sequencing and were assigned to 10 different actinomycete genera. Eight putatively novel species belonging to genera Kocuria, Mycobacterium, Nocardia, and Rhodococcus were identified based on sequence similarity values below 98.2% to other 16S rRNA gene sequences available in the NCBI database. PCR-based screening for biosynthetic genes including type I and type II polyketide synthases (PKS-I, PKS-II as well as nonribosomal peptide synthetases (NRPS showed that 20 actinomycete isolates encoded each at least one type of biosynthetic gene. The organic extracts of nine isolates displayed bioactivity against at least one of the test pathogens, which were Gram-positive and Gram-negative bacteria, fungi, human parasites, as well as in a West Nile Virus protease enzymatic assay. These results emphasize that marine sponges are a prolific resource for novel bioactive actinomycetes with potential for drug discovery.

  14. Actinomycetes from the South China Sea sponges: isolation, diversity, and potential for aromatic polyketides discovery

    Science.gov (United States)

    Sun, Wei; Zhang, Fengli; He, Liming; Karthik, Loganathan; Li, Zhiyong

    2015-01-01

    Marine sponges often harbor dense and diverse microbial communities including actinobacteria. To date no comprehensive investigation has been performed on the culturable diversity of the actinomycetes associated with South China Sea sponges. Structurally novel aromatic polyketides were recently discovered from marine sponge-derived Streptomyces and Saccharopolyspora strains, suggesting that sponge-associated actinomycetes can serve as a new source of aromatic polyketides. In this study, a total of 77 actinomycete strains were isolated from 15 South China Sea sponge species. Phylogenetic characterization of the isolates based on 16S rRNA gene sequencing supported their assignment to 12 families and 20 genera, among which three rare genera (Marihabitans, Polymorphospora, and Streptomonospora) were isolated from marine sponges for the first time. Subsequently, β-ketoacyl synthase (KSα) gene was used as marker for evaluating the potential of the actinomycete strains to produce aromatic polyketides. As a result, KSα gene was detected in 35 isolates related to seven genera (Kocuria, Micromonospora, Nocardia, Nocardiopsis, Saccharopolyspora, Salinispora, and Streptomyces). Finally, 10 strains were selected for small-scale fermentation, and one angucycline compound was detected from the culture extract of Streptomyces anulatus strain S71. This study advanced our knowledge of the sponge-associated actinomycetes regarding their diversity and potential in producing aromatic polyketides. PMID:26483773

  15. Isolation, phylogenetic analysis and anti-infective activity screening of marine sponge-associated actinomycetes.

    Science.gov (United States)

    Abdelmohsen, Usama Ramadan; Pimentel-Elardo, Sheila M; Hanora, Amro; Radwan, Mona; Abou-El-Ela, Soad H; Ahmed, Safwat; Hentschel, Ute

    2010-02-26

    Terrestrial actinomycetes are noteworthy producers of a multitude of antibiotics, however the marine representatives are much less studied in this regard. In this study, 90 actinomycetes were isolated from 11 different species of marine sponges that had been collected from offshore Ras Mohamed (Egypt) and from Rovinj (Croatia). Phylogenetic characterization of the isolates based on 16S rRNA gene sequencing supported their assignment to 18 different actinomycete genera representing seven different suborders. Fourteen putatively novel species were identified based on sequence similarity values below 98.2% to other strains in the NCBI database. A putative new genus related to Rubrobacter was isolated on M1 agar that had been amended with sponge extract, thus highlighting the need for innovative cultivation protocols. Testing for anti-infective activities was performed against clinically relevant, Gram-positive (Enterococcus faecalis, Staphylococcus aureus) and Gram-negative (Escherichia coli, Pseudomonas aeruginosa) bacteria, fungi (Candida albicans) and human parasites (Leishmania major, Trypanosoma brucei). Bioactivities against these pathogens were documented for 10 actinomycete isolates. These results show a high diversity of actinomycetes associated with marine sponges as well as highlight their potential to produce anti-infective agents.

  16. Isolation, Phylogenetic Analysis and Anti-infective Activity Screening of Marine Sponge-Associated Actinomycetes

    Directory of Open Access Journals (Sweden)

    Safwat Ahmed

    2010-02-01

    Full Text Available Terrestrial actinomycetes are noteworthy producers of a multitude of antibiotics, however the marine representatives are much less studied in this regard. In this study, 90 actinomycetes were isolated from 11 different species of marine sponges that had been collected from offshore Ras Mohamed (Egypt and from Rovinj (Croatia. Phylogenetic characterization of the isolates based on 16S rRNA gene sequencing supported their assignment to 18 different actinomycete genera representing seven different suborders. Fourteen putatively novel species were identified based on sequence similarity values below 98.2% to other strains in the NCBI database. A putative new genus related to Rubrobacter was isolated on M1 agar that had been amended with sponge extract, thus highlighting the need for innovative cultivation protocols. Testing for anti-infective activities was performed against clinically relevant, Gram-positive (Enterococcus faecalis, Staphylococcus aureus and Gram-negative (Escherichia coli, Pseudomonas aeruginosa bacteria, fungi (Candida albicans and human parasites (Leishmania major, Trypanosoma brucei. Bioactivities against these pathogens were documented for 10 actinomycete isolates. These results show a high diversity of actinomycetes associated with marine sponges as well as highlight their potential to produce anti-infective agents.

  17. Actinomycetes for marine drug discovery isolated from mangrove soils and plants in China.

    Science.gov (United States)

    Hong, Kui; Gao, An-Hui; Xie, Qing-Yi; Gao, Hao; Zhuang, Ling; Lin, Hai-Peng; Yu, Hai-Ping; Li, Jia; Yao, Xin-Sheng; Goodfellow, Michael; Ruan, Ji-Sheng

    2009-01-01

    The mangrove ecosystem is a largely unexplored source for actinomycetes with the potential to produce biologically active secondary metabolites. Consequently, we set out to isolate, characterize and screen actinomycetes from soil and plant material collected from eight mangrove sites in China. Over 2,000 actinomycetes were isolated and of these approximately 20%, 5%, and 10% inhibited the growth of Human Colon Tumor 116 cells, Candida albicans and Staphylococcus aureus, respectively, while 3% inhibited protein tyrosine phosphatase 1B (PTP1B), a protein related to diabetes. In addition, nine isolates inhibited aurora kinase A, an anti-cancer related protein, and three inhibited caspase 3, a protein related to neurodegenerative diseases. Representative bioactive isolates were characterized using genotypic and phenotypic procedures and classified to thirteen genera, notably to the genera Micromonospora and Streptomyces. Actinomycetes showing cytotoxic activity were assigned to seven genera whereas only Micromonospora and Streptomyces strains showed anti-PTP1B activity. We conclude that actinomycetes isolated from mangrove habitats are a potentially rich source for the discovery of anti-infection and anti-tumor compounds, and of agents for treating neurodegenerative diseases and diabetes.

  18. Isolation and characterization of marine-derived actinomycetes with cytotoxic activity from the Red Sea coast

    Institute of Scientific and Technical Information of China (English)

    Mohamed Saleh Abdelfattah; Mohammed Ismail Youssef Elmallah; Usama Wahid Hawas; Lamia Taha Abou El-Kassema; Mennat Allah Gamal Eid

    2016-01-01

    Objective: To isolate and evaluate the cytotoxic activity of different actinomycetes species isolated from the Red Sea coast in Sharm el-Sheikh, Egypt.Methods: Forty actinomycetes strains were isolated from different sediments and seawater samples collected from the Red Sea coast in Egypt. Actinomycetes were recognized by morphological and microscopic examinations. Cell viability and cytotoxicity induced by the crude extracts on breast cancer cell lines MDA-MB-231 were assessed using methylene blue assay. The strains with promising cytotoxic activity were identified by sequencing and amplifying the 16 S r RNA genes. The antibacterial activities of the crude extracts were performed using Kirby–Bauer disc diffusion method.Results: The results indicated that five ethyl acetate extracts exhibited cytotoxicity towards breast cancer cell lines MDA-MB-231. The highest cytotoxic activity was found for the ethyl acetate extracts of EGY2 and EGY39. The isolate EGY3 was identified as a new Streptomyces species, while the actinomycete EGY22 was found to be a member of the genus Nocardiopsis sp. The crude extract of the isolate EGY8 showed slightly high antimicrobial activity against different test microorganisms.Conclusions: The results of the present study reveal that marine sediments of the Red Sea are a potent source of novel species of actinomycetes. The isolates may be useful in discovery of novel bioactive compounds and an important step in the development of microbial natural product research.

  19. Isolation of actinomycetes from mangrove and estuarine sediments of Cochin and screening for antimicrobial activity

    Institute of Scientific and Technical Information of China (English)

    Emilda Rosmine

    2016-01-01

    Objective:To isolate and screen actinomycetes for antimicrobial activity from mangroves and estuarine soil samples of Cochin. Methods: In the present study, sediment samples collected from mangroves and various stations of Cochin estuary were pretreated and actinomycetes were isolated on different selective media. The isolates were screened for antibiotic activity by following disc diffusion assay (Kirby-Bauer method) against human pathogens, fish pathogens and Gram-positive bacteria. The isolates were identified based on their morphology. Results:Only 2 actinomycete isolates (ER7and ER10) of the 50 isolates screened had antimicrobial activities against one or more pathogens tested. ER7 isolate showed higher antimicrobial activity as compared to that of ER10 isolate. The maximum inhibition zone of crude extract from ER7 was 16.7 mm. The methanol extract of ER7 showed antimicrobial activity against all the pathogens tested with a maximum zone of 21.0 mm. The isolates with antimicrobial activity were found to belong to the genusStreptomyces. Conclusions:There is no significant report on bioactive actinomycetes from the present study areas. Potent antibiotics from the selected isolates could contribute to fight against several human and fish diseases. Further purification, structural elucidation and characterization are recommended to know the quality, novelty and commercial value of these antibiotics. Hence, the mangroves and estuary of Kochi show great promise for the discovery of bioactive actinomycetes.

  20. Actinomycetes from red sea sponges: Sources for chemical and phylogenetic diversity

    KAUST Repository

    Abdelmohsen, Usama Ramadan

    2014-05-12

    The diversity of actinomycetes associated with marine sponges collected off Fsar Reef (Saudi Arabia) was investigated in the present study. Forty-seven actinomycetes were cultivated and phylogenetically identified based on 16S rRNA gene sequencing and were assigned to 10 different actinomycete genera. Eight putatively novel species belonging to genera Kocuria, Mycobacterium, Nocardia, and Rhodococcus were identified based on sequence similarity values below 98.2% to other 16S rRNA gene sequences available in the NCBI database. PCR-based screening for biosynthetic genes including type I and type II polyketide synthases (PKS-I, PKS-II) as well as nonribosomal peptide synthetases (NRPS) showed that 20 actinomycete isolates encoded each at least one type of biosynthetic gene. The organic extracts of nine isolates displayed bioactivity against at least one of the test pathogens, which were Gram-positive and Gram-negative bacteria, fungi, human parasites, as well as in a West Nile Virus protease enzymatic assay. These results emphasize that marine sponges are a prolific resource for novel bioactive actinomycetes with potential for drug discovery. 2014 by the authors; licensee MDPI.

  1. lsolation and characterization of marine-derived actinomycetes with cytotoxic activity from the Red Sea coast

    Institute of Scientific and Technical Information of China (English)

    Mohamed Saleh Abdelfattah; Usama Wahid Hawas; Lamia Taha Abou El-Kassema; Mennat Allah Gamal Eid

    2016-01-01

    Objective: To isolate and evaluate the cytotoxic activity of different actinomycetes species isolated from the Red Sea coast in Sharm el-Sheikh, Egypt. Methods: Forty actinomycetes strains were isolated from different sediments and seawater samples collected from the Red Sea coast in Egypt. Actinomycetes were recognized by morphological and microscopic examinations. Cell viability and cyto-toxicity induced by the crude extracts on breast cancer cell lines MDA-MB-231 were assessed using methylene blue assay. The strains with promising cytotoxic activity were identified by sequencing and amplifying the 16S rRNA genes. The antibacterial activities of the crude extracts were performed using Kirby-Bauer disc diffusion method. Results: The results indicated that five ethyl acetate extracts exhibited cytotoxicity to-wards breast cancer cell lines MDA-MB-231. The highest cytotoxic activity was found for the ethyl acetate extracts of EGY2 and EGY39. The isolate EGY3 was identified as a new Streptomyces species, while the actinomycete EGY22 was found to be a member of the genus Nocardiopsis sp. The crude extract of the isolate EGY8 showed slightly high antimicrobial activity against different test microorganisms. Conclusions: The results of the present study reveal that marine sediments of the Red Sea are a potent source of novel species of actinomycetes. The isolates may be useful in discovery of novel bioactive compounds and an important step in the development of microbial natural product research.

  2. SCREENING OF ANTIMICROBIAL ACTIVITY AND GENES CODING POLYKETIDE SYNTHETASE AND NONRIBOSOMAL PEPTIDE SYNTHETASE OF ACTINOMYCETE ISOLATES

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    Silvia Kovácsová

    2013-12-01

    Full Text Available The aim of this study was to observe antimicrobial activity using agar plate diffusion method and screening genes coding polyketide synthetase (PKS-I and nonribosomal peptide synthetase (NRPS from actinomycetes. A total of 105 actinomycete strains were isolated from arable soil. Antimicrobial activity was demonstrated at 54 strains against at least 1 of total 12 indicator organisms. Antifungal properties were recorded more often than antibacterial properties. The presence of PKS-I and NRPS genes were founded at 61 of total 105 strains. The number of strains with mentioned biosynthetic enzyme gene fragments matching the anticipated length were 19 (18% and 50 (47% respectively. Overall, five actinomycete strains carried all the biosynthetical genes, yet no antimicrobial activity was found against any of tested pathogens. On the other hand, twenty-one strains showed antimicrobial activity even though we were not able to amplify any of the PKS or NRPS genes from them. Combination of the two methods showed broad-spectrum antimicrobial activity of actinomycetes isolated from arable soil, which indicate that actinomycetes are valuable reservoirs of novel bioactive compounds.

  3. Screening of Actinomycetes From Lipar Area of Oman Sea to Investigate the Antibacterial Compounds

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    Shams

    2015-02-01

    Full Text Available Background Actinomycetes are one of the most important sources for the production of antibacterial compounds. Marine environments, due to their unique characteristics, are considered a good option to search for bacteria with the capability of producing antimicrobial compounds. Objectives The purpose of this study was to isolate the actinomycetes producing antibacterial compounds. Materials and Methods A total of 35 actinomycetes were isolated from Oman Sea (Lipar Area. To investigate antibacterial activity, the isolated actinomycetes were assessed against reference and pathogenic bacteria, including Staphylococcus epidermidis, Staphylococcu intermedius, Staphylococcu chromogenes, Staphylococcu saprophyticus, Bacillus cereus and methicillin-resistance Staphylococcu aureus, Pseudomonas, Listeria, Klebsiella, Salmonella, Acinetobacter, and Escherichia coli O157:H7, using the cross streak method. Results Based on the morphological characterization, 35 isolated cases belonged to actinomycetes and %94 of them had the ability to produce antibacterial compounds. In the cross streak method, most of the isolated bacteria have antibacterial activity against reference S. aureus among Gram-positive bacteria and Acinetobacter among Gram-negative bacteria. Inhibition zone diameters were measured between 2-25 and 1-20 mm for Gram-positive and -negative bacteria, receptivity. Conclusions Preliminary results indicate that the native Iranian Actinobacteria could be considered a suitable option for screening of the new antibacterial compounds. Molecular research and antibacterial compound extraction against the aforementioned pathogenic strains are also being conducted.

  4. Amylase activity of aquatic actinomycetes isolated from the sediments of mangrove forests in south of Iran

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    Farshid Kafilzadeh

    2015-01-01

    Full Text Available In this study amylase producing actinomycetes were isolated from the sediments of mangrove forests in the south of Iran and the rate of amylase activity was measured. The samples of sediments were collected from one hundred different places in mangrove forests of the south of Iran. Collected samples were diluted then they were purified on the starch (casein agar culture and Woodruff. After that they were examined in terms of amylase production on agar–starch culture. The activity of the produced amylase by the isolated aquatic actinomycetes was measured by dinitrosalicylic acid (DNS method. The results showed that aquatic actinomycetes were isolated from 86 per 100 places in spring (86% and from 61 per 100 places in summer (61%. The highest rates of producing enzyme were related to isolated samples in spring (62.97 U/ml. Biochemical and Bergey’s book tests showed that the most isolated aquatic actinomycetes belonged to Streptomyces genus. As regards this, it is economical and easy to isolate the aquatic actinomycetes which produce amylase that is used in different industries in Iran from the sediments of mangrove forests of the south of Iran. So the isolated strains in this study can be suitable candidates for amylase production after genetic manipulation.

  5. Phylogenetic characterization of culturable actinomycetes associated with the mucus of the coral Acropora digitifera from Gulf of Mannar.

    Science.gov (United States)

    Nithyanand, Paramasivam; Manju, Sivalingam; Karutha Pandian, Shunmugiah

    2011-01-01

    The marine environment is a virtually untapped source of novel actinomycete diversity and its metabolites. Investigating the diversity of actinomycetes in other marine macroorganisms, like seaweeds and sponges, have resulted in isolation of novel bioactive metabolites. Actinomycetes diversity associated with corals and their produced metabolites have not yet been explored. Hence, in this study we attempted to characterize the culturable actinomycetes population associated with the coral Acropora digitifera. Actinomycetes were isolated from the mucus of the coral wherein the actinomycetes count was much higher when compared with the surrounding seawater and sediment. Actinobacteria-specific 16S rRNA gene primers were used for identifying the isolates at the molecular level in addition to biochemical tests. Amplified ribosomal DNA restriction analysis using three restriction enzymes revealed several polymorphic groups within the isolates. Sequencing and blast analysis of the isolates revealed that some isolates had only 96.7% similarity with its nearest match in GenBank indicating that they may be novel isolates at the species level. The isolated actinomycetes exhibited good antibacterial activity against various human pathogens. This study offers for the first time a prelude about the unexplored culturable actinomycetes diversity associated with a scleractinian coral and their bioactive capabilities. © 2010 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  6. Complete genome sequence of Desulfurococcus fermentans, a hyperthermophilic cellulolytic crenarchaeon isolated from a freshwater hot spring in Kamchatka, Russia.

    Science.gov (United States)

    Susanti, Dwi; Johnson, Eric F; Rodriguez, Jason R; Anderson, Iain; Perevalova, Anna A; Kyrpides, Nikos; Lucas, Susan; Han, James; Lapidus, Alla; Cheng, Jan-Fang; Goodwin, Lynne; Pitluck, Sam; Mavrommatis, Konstantinos; Peters, Lin; Land, Miriam L; Hauser, Loren; Gopalan, Venkat; Chan, Patricia P; Lowe, Todd M; Atomi, Haruyuki; Bonch-Osmolovskaya, Elizaveta A; Woyke, Tanja; Mukhopadhyay, Biswarup

    2012-10-01

    Desulfurococcus fermentans is the first known cellulolytic archaeon. This hyperthermophilic and strictly anaerobic crenarchaeon produces hydrogen from fermentation of various carbohydrates and peptides without inhibition by accumulating hydrogen. The complete genome sequence reported here suggested that D. fermentans employs membrane-bound hydrogenases and novel glycohydrolases for hydrogen production from cellulose.

  7. Complete Genome Sequence of Desulfurococcus fermentans, a Hyperthermophilic Cellulolytic Crenarchaeon Isolated from a Freshwater Hot Spring in Kamchatka, Russia

    Energy Technology Data Exchange (ETDEWEB)

    Susanti, Dwi [Virginia Polytechnic Institute and State University (Virginia Tech); Johnson, Eric F [Virginia Polytechnic Institute and State University (Virginia Tech); Rodriquez, Jason [Virginia Polytechnic Institute and State University (Virginia Tech); Anderson, Iain [U.S. Department of Energy, Joint Genome Institute; Perevalova, Anna [Virginia Polytechnic Institute and State University (Virginia Tech); Kyrpides, Nikos C [U.S. Department of Energy, Joint Genome Institute; Lucas, Susan [U.S. Department of Energy, Joint Genome Institute; Han, James [U.S. Department of Energy, Joint Genome Institute; Lapidus, Alla L. [U.S. Department of Energy, Joint Genome Institute; Cheng, Jan-Fang [U.S. Department of Energy, Joint Genome Institute; Goodwin, Lynne A. [Los Alamos National Laboratory (LANL); Pitluck, Sam [U.S. Department of Energy, Joint Genome Institute; Mavromatis, K [U.S. Department of Energy, Joint Genome Institute; Peters, Lin [U.S. Department of Energy, Joint Genome Institute; Land, Miriam L [ORNL; Hauser, Loren John [ORNL; Gopapan, Venkay [Ohio State University; Chan, Patricia [University of California, Santa Cruz; Atomi, Haruyuki [Kyoto University, Japan; Bonch-Osmolovskaya, Elizaveta [Russian Academy of Sciences, Moscow; Woyke, Tanja [U.S. Department of Energy, Joint Genome Institute; Mukhopadhyay, Biswarup [Virginia Polytechnic Institute and State University (Virginia Tech)

    2012-01-01

    Desulfurococcus fermentans is the first known cellulolytic archaeon. This hyperthermophilic and strictly anaerobic crenarchaeon produces hydrogen from fermentation of various carbohydrates and peptides without inhibition by accumulating hydrogen. The complete genome sequence reported here suggested that D. fermentans employs membrane-bound hydrogenases and novel glycohydrolases for hydrogen production from cellulose.

  8. Screening of secondary metabolite biosynthesis genes of marine actinomycetes isolated from Trabzon (Black Sea sea sediments

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    Kadriye Özcan

    2017-06-01

    Full Text Available In this study, active secondary metabolite production capacity of actinomycete isolates obtained from Trabzon (Black Sea sea sediments was investigated by molecular techniques. Totaly 24 actinomycetes were investigated by PCR based on the presence of secondary metabolite biosynthesis genes PKS / NRPS. According to the PCR results, 25 and 58% of actinomycetes obtained from Trabzon sea sediments were found to contain PKS-NRPS and only NRPS gene regions, respectively. When PCR data were evaluated, it was found that the production of the peptide form active secondary metabolite of the isolates by non-ribosomal way was higher than that of the secondary metabolite production by the PKS pathway. In addition, it has been determined that Black Sea marine sediments have high potential for active secondary metabolite production.

  9. Harnessing the Potential of Halogenated Natural Product Biosynthesis by Mangrove-Derived Actinomycetes

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    Xiang Xiao

    2013-10-01

    Full Text Available Mangrove-derived actinomycetes are promising sources of bioactive natural products. In this study, using homologous screening of the biosynthetic genes and anti-microorganism/tumor assaying, 163 strains of actinomycetes isolated from mangrove sediments were investigated for their potential to produce halogenated metabolites. The FADH2-dependent halogenase genes, identified in PCR-screening, were clustered in distinct clades in the phylogenetic analysis. The coexistence of either polyketide synthase (PKS or nonribosomal peptide synthetase (NRPS as the backbone synthetases in the strains harboring the halogenase indicated that these strains had the potential to produce structurally diversified antibiotics. As a validation, a new enduracidin producer, Streptomyces atrovirens MGR140, was identified and confirmed by gene disruption and HPLC analysis. Moreover, a putative ansamycin biosynthesis gene cluster was detected in Streptomyces albogriseolus MGR072. Our results highlight that combined genome mining is an efficient technique to tap promising sources of halogenated natural products synthesized by mangrove-derived actinomycetes.

  10. Actinomycetal complexes in drained peat soils of the taiga zone upon pyrogenic succession

    Science.gov (United States)

    Zenova, G. M.; Glushkova, N. A.; Bannikov, M. V.; Shvarov, A. P.; Pozdnyakov, A. I.; Zvyagintsev, D. G.

    2008-04-01

    The number and diversity of actinomycetes in peat soils vary in dependence on the stage of pyrogenic succession. In the cultivated peat soil, the number of actinomycetes after fires decreases by three-four times, mainly at the expense of acidophilic and neutrophilic groups. An increase in the number of mycelial prokaryotes (at the expense of alkaliphilic forms) is seen on the fifth year of functioning of the pyrogenic peat soil. The species diversity of streptomycetes in peat soils also decreases after fires. An increase in the range of streptomycetal species at the expense of neutrophilic and alkaliphilic forms takes place on the fifth year of the pyrogenic succession. Parameters of the actinomycetal complex—the population density, species composition, and ecological features—are the criteria whose changes allow us to judge the state of peat soils in the course of their pyrogenic succession.

  11. A comparative study on selected marine actinomycetes from Pulicat, Muttukadu, and Ennore estuaries

    Institute of Scientific and Technical Information of China (English)

    SChacko Vijai Sharma; Ernest David

    2012-01-01

    Objective: To isolate and make a comparative study of marine sediments actinomycetes from Pulicat estuary, Muttukadu estuary and Ennore estuary, TamilNadu, India. Methods: A unique selective enrichment procedure has resulted in the isolation and identification a total of 304 actinomycetes colonies which were isolated from different stations of marine soil sediments in Pulicat estuary, Muttukadu estuary and Ennore estuary, TamilNadu, India. Results: Among them, 277 isolates were morphologically distinct on the basis of spore mass colour, aerial and substrate mycelium formation and production of diffusible pigment. The majority (60%; 162 isolates) were assigned to the genus Streptomyces. (35%; 104 isolates) were assigned to the genus Actinopolyspora, (5%; 11 isolates) were assigned to the genus Nocardiodes. Conclusions: The present study concluded that the physiological characteristics of actinomycetes Streptomyces, Actinopolyspora and Nocardiodes varied by available nutrients in the medium and the physical conditions.

  12. Exploration of Potential Actinomycetes from CIFOR Forest Origin as Antimicrobial, Antifungus, and Producing Extracellular Xylanase

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    Sipriyadi Sipriyadi

    2016-03-01

    Full Text Available This study aimed to isolate and explore the actinomycetes of CIFOR forest origin as an antimicrobial and antifungal agent, to produce an extracellular xylanase, and to identify isolates based on 16S rRNA gene sequences. Actinomycetes were isolated using Humic-acid Vitamin-B agar (HV media. Actinomycetes colonies that grow on the medium HV was subsequently purified by growing them on yeast malt agar (YMA media, then an antagonistic test of selected bacteria against Bacillus sp., Escherichia coli, Fusarium oxysporum, and Sclerotium sp was performed. Xylanase activity test was detected by observing a clear zone, followed by identification. Total of 35 isolates of actinomycetes isolated based on their colony morphology characteristics and diverse types of spore chains showed Streptomyces spp. of isolates CFR-06, CFR-15, CFR-17, CFR-18, and CFR-19 were able to inhibit the growth of Bacillus sp.. The highest inhibition zone has a diameter of 10.1 mm (isolate CFR-17. Isolates CFR-01 and CFR-15 were able to inhibit the growth of E. coli with the highest inhibition zone diameter of 5.1 mm (isolate CFR-15. Isolates CFR-29 and CFR-12 were able to inhibit the growth of F. oxysporum while isolate CFR-35 were able to inhibit the growth of Sclerotium sp.. Xylanase activity test showed that isolates CFR-12, CFR-20, CFR-22, CFR-24, CFR-25, CFR-30, CFR-33, CFR-34 have an ability to produce extracellular xylanase enzyme. Actinomycetes isolate (Xyl_22 as a potential xylanase enzyme producer was closely related with Streptomyces drozdowicii by the maximum similarity of 99%.How to CiteSipriyadi, S., Lestari, Y., Wahyudi, A., Meryandini, A., & Suhartono, M. T. (2016. Exploration Potential CIFOR Forest actinomycetes origin as Antimicrobial, Anti Fungus and Producing Enzymes Extracellular Xylanase. Biosaintifika: Journal of Biology & Biology Education, 8(1, 94-102.

  13. [Analysis of the halogenase gene in actinomycetes from different habitats and its implications for halometabolite discovery].

    Science.gov (United States)

    Gao, Peng; Xi, Lijun; Piao, Yuhua; Ruan, Jisheng; Huang, Ying

    2009-10-01

    To compare the halometabolite producing capability between actinomycetes of earth origin and marine origin, based on genetic screening of the 1,5-dihydroflavin adenine dinucleotide (FADH2-dependent) halogenase gene. We used 141 actinomycete isolates that were dereplicated by phenotype, 70 of earth origin and 71 of marine origin, and obtained halogenase gene fragments from them by PCR screening. We then sequenced the PCR products and analyzed corresponding amino acid sequences phylogenetically. We made further comparison of the halogenase sequences between actinomycetes of different origins, and between marine-origin streptomycetes and marine-origin Micromonospora isolates. In addition, we detected polyketide synthase (PKS) and non-ribosomal peptide synthetase (NRPS) genes by PCR in the halogenase gene-positive isolates. We observed higher occurrence of the halogenase gene in marine-origin actinomycetes (36.6%) than in earth-origin actinomycetes (14.3%), and in marine-origin streptomycetes (69.0%) than in marine-origin Micromonospora isolates (14.3%). Most (86.1%) of the halogenase gene-positive isolates contained PKS and/or NRPS genes. Moreover, the halogenase sequences of marine-origin isolates differed largely from the known ones, and clustered into a couple of distinct clades in the phylogenetic tree. In addition, we found greater diversity of the halogenase genes in marine-origin Micromonospora isolates than in marine-origin streptomycetes. Based on the results of this study, we propose that actinomycetes, especially streptomycetes, from marine habitat could serve as a good source for new bioactive halometabolite discovery in the future.

  14. Antibacterial activity of actinomycetes isolated from different soil samples of Sheopur (A city of central India

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    Hotam S Chaudhary

    2013-01-01

    Full Text Available The main objective of the present study was isolation, purification, and characterization of actinomycetes from soil samples, having antimicrobial activity against 12 selected pathogenic strains. Soils samples were taken from different niche habitats of Sheopur district, Madhya Pradesh, India. These samples were serially diluted and plated on actinomycete isolation agar media. Potential colonies were screened, purified, and stored in glycerol stock. Isolates were morphologically and biochemically characterized. These isolates were subjected to extraction for production of the antibacterial compound. Antibacterial activity and Minimum Inhibitory Concentration (MIC of the purified extract of isolates were evaluated. Totally 31 actinomycete isolates were tested for antagonistic activity against 12 pathogenic microorganisms. Isolates AS14, AS27, and AS28 were highly active, while AS1 showed less activity against the pathogenic microorganisms. Isolate AS7 exhibited the highest antagonistic activity against Bacillus cereus (24 mm and AS16 showed the highest activity against Enterococcus faecalis (21 mm. MIC was also determined for actinomycete isolates against all the tested microorganisms. MIC of actinomycete isolates was found to be 2.5 mg/ml against Shigella dysenteriae, Vancomycin-resistant enterococci, and Klebsiella pneumoniae, and was 1.25 mg/ml for Staphylococcus saprophyticus, Streptococcus pyogenes, Staphylococcus epidermidis, Methicillin-resistant Staphylococcus, Bacillus cereus, Staphylococcus xylosus, Methicillin-resistant Staphylococcus aureus, Enterococcus faecalis, and Staphylococcus aureus. All actinomycetes isolates showed antibacterial activity against S. aureus, while they showed less activity against S. dysenteriae. These isolates had antibacterial activity and could be used in the development of new antibiotics for pharmaceutical or agricultural purposes.

  15. Comprehensive analysis of the cellulolytic system reveals its potential for deconstruction of lignocellulosic biomass in a novel Streptomyces sp.

    Science.gov (United States)

    Pinheiro, Guilherme L; de Azevedo-Martins, Allan C; Albano, Rodolpho M; de Souza, Wanderley; Frases, Susana

    2017-01-01

    The giant snail Achatina fulica is considered an invasive species in most territories in which it was introduced, due to its ability to process a large amount of lignocellulose as a consequence of the presence of a cellulolytic-associated microflora. Streptomyces are well known as crucial agents in the decomposition of complex polymers in soil environments and also as cellulolytic symbionts commonly associated with herbivore insects. Here, we employed a combination of genomic and biochemical tools for a detailed evaluation of the cellulolytic potential of Streptomyces sp. I1.2, an aerobic bacterium isolated from the intestinal lumen of A. fulica in a screening for cellulolytic bacteria. Genomic analysis revealed that the ratio and diversity of CAZy domains and GH families coded by Streptomyces sp. I1.2 are comparable to those present in other highly cellulolytic bacteria. After growth on crystalline cellulose or sugarcane bagasse as sole carbon sources, the functionality of several genes encoding endoglucanases, cellobiohydrolases, xylanases, CBMs, and one β-glucosidase were confirmed by the combination of enzymatic activity measurements, zymography, TLC, and cellulose-binding assays. The endoglucanases secreted by this isolate were stable at 50 °C and exhibited activity over a broad pH range between 4.0 and 8.0. The endoglucanases and cellobiohydrolases secreted by Streptomyces sp. I1.2 exhibited specific activities that were similar to the levels present in a commercial cellulase preparation from Trichoderma reesei, while I1.2 xylanase levels were even 350 % higher. The results presented here show that Streptomyces sp. I1.2 is promising for future biotechnological applications, since it is able to produce endoglucanases, cellobiohydrolases, and xylanases in appreciable amounts when grown on a low-cost residue such as sugarcane bagasse.

  16. Actinomycetes bioactivos de sedimento marino de la costa central del Perú

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    Jorge León

    2013-04-01

    Full Text Available En el presente trabajo evaluamos la actividad antibacteriana y antifúngica de actinomycetes marinos sobre patógenos de origen clínico. Asimismo, fueron evaluadas la capacidad de producir enzimas extracelulares como carbohidrasas, lipasas y proteasas. Los Actinomycetes fueron aislados de sedimentos colectados entre setiembre a diciembre del 2005 de las Bahías de Ancón (Lima e Independencia (Ica de 34 y 100 m de profundidad. El aislamiento se realizó en Agar Caseína - Almidón (ACA y Agar Marino (AM con adición de Cicloheximide (10 μg/mL. Las evaluaciones antimicrobianas fueron realizadas frente a bacterias patógenas antibiótico-multirresistentes y hongos de origen clínico; en tanto, para evaluar su actividad multienzimática se utilizaron sustratos poliméricos diversos. Se aislaron un total de 62 actinomycetes, de los cuales 31 (50% mostraron actividad antibacteriana frente a Staphylococcus aureus, 36 (59% frente a Pseudomonas aeruginosa y 23 (37% a ambos patógenos. Las cepas de actinomycetes I-400A y M10-77 identificadas en cada caso como Streptomyces y Thermoactinomyces fueron las que exhibieron mayor actividad inhibitoria frente a P. aeruginosa y S. aureus respectivamente. Asimismo, 13 actinomycetes (20,97% mostraron actividad antifúngica frente a cultivos de Candida albicans cepa 1511 y 17 (27,42% frente a Candida albicans cepa 1511MIC; sin embargo, ningún actinomycete presentó actividad inhibitoria frente a Aspergillus niger, Aspergillus fumigatus y Trichophyton mentagrophytes. La mayoría de los actinomycetes mostraron tener actividad multienzimática capaz de hidrolizar compuestos poliméricos como el tween-80 (96%, la gelatina (95%, almidón (93%, lecitina (88% y la caseína (74%. Extractos del compuesto activo obtenidos de la cepa M10-77 con etil acetato rindieron notable actividad inhibitoria contra S. aureus. Se concluye que el sedimento marino es fuente de Actinomycetes con gran capacidad de producir sustancias

  17. Actinomycetes bioactivos de sedimento marino de la costa central del Perú

    OpenAIRE

    Jorge León; Libia Liza; Isela Soto; D´Lourdes Cuadra; Lilian Patiño; Rito Zerpa

    2013-01-01

    En el presente trabajo evaluamos la actividad antibacteriana y antifúngica de actinomycetes marinos sobre patógenos de origen clínico. Asimismo, fueron evaluadas la capacidad de producir enzimas extracelulares como carbohidrasas, lipasas y proteasas. Los Actinomycetes fueron aislados de sedimentos colectados entre setiembre a diciembre del 2005 de las Bahías de Ancón (Lima) e Independencia (Ica) de 34 y 100 m de profundidad. El aislamiento se realizó en Agar Caseína - Almidón (ACA) y Agar Mar...

  18. Glucose metabolism in the antibiotic producing actinomycete Nonomuraea sp ATCC 39727

    DEFF Research Database (Denmark)

    Gunnarsson, Nina; Bruheim, Per; Nielsen, Jens

    2004-01-01

    The actinomycete Nonomuraea sp. ATCC 39727, producer of the glycopeptide A40926 that is used as precursor for the novel antibiotic dalbavancin, has an unusual carbon metabolism. Glucose is primarily metabolized via the Entner-Doudoroff (ED) pathway, although the energetically more favorable Embden...... - Meyerhof - Parnas (EMP) pathway is present in this organism. Moreover, Nonomuraea utilizes a PPi-dependent phosphofructokinase, an enzyme that has been connected with anaerobic metabolism in eukaryotes and higher plants, but recently has been recognized in several actinomycetes. In order to study its...

  19. Production of cellulolytic enzymes by Pleurotus species on lignocellulosic wastes using novel pretreatments.

    Science.gov (United States)

    Singh, M P; Pandey, A K; Vishwakarma, S K; Srivastava, A K; Pandey, V K; Singh, V K

    2014-12-24

    In the present investigation three species of Pleurotus i.e. P. sajor—caju (P1), P. florida (P2) and P. flabellatus (P3) along with two lignocellulosic substrates namely paddy straw and wheat straw were selected for evaluation of production of extracellular cellulolytic enzymes. During the cultivation of three species of Pleurotus under in vivo condition, the two lignocellulosic substrates were treated with plants extracts (aqueous extracts of ashoka leaves (A) and neem oil (B)), hot water (H) and chemicals (C).Among all treatments, neem oil treated substrates supported better enzyme production followed by aqueous extract of ashoka leaves, hot water and chemical treatment. Between the two substrates paddy straw supported better enzyme production than wheat straw. P. flabellatus showed maximum activity of exoglucanase, endoglucanase and β—glucosidase followed by P. florida and P. sajor—caju.

  20. Degradation and mineralization of wheat straw by some cellulolytic fungi in pure cultures

    Energy Technology Data Exchange (ETDEWEB)

    Omar, S.A. (Botany Dept., Faculty of Science, Assiut Univ. (Egypt))

    1994-01-01

    Decomposition and mineralization of wheat straw inoculated with 8 cellulolytic fungi was investigated in sand culture. Aspergillus fumigatus and Stachybotrys chartarum showed a great potential to degrade wheat straw and weight loss of straw was 35.3 and 27.5% of initial weight after 63 days incubation, respectively. CO[sub 2] production was highest after 7 and 14 days for all tested fungi suggesting that the main contribution to CO[sub 2] production arose from the water-soluble fraction which was decomposed rapidly. Carbon solubilization from wheat straw occurred, to some extent, in a manner similar to that of straw decomposition. Also, S. chartarum and A. fumigatus solubilized considerable amounts of straw Nitrogen but Gibberella fujikuroi was the best nitrogen dissolving organism. (orig.)

  1. In vitro cellulolytic activity of the plant pathogen Clavibacter michiganensis subsp. sepedonicus.

    Science.gov (United States)

    Baer, D; Gudmestad, N C

    1995-10-01

    The activity of four Clavibacter michiganensis subsp. sepedonicus strains against various cellulose substrates was investigated. Sixty-seven Clavibacter michiganensis subsp. sepedonicus strains grew well on media amended with carboxymethylcellulose, 64 strains produced zones of hydrolysis. Endoglucanase activity was optimal at 37 degrees C and pH 6.0 against carboxymethylcellulose incorporated in plate assays. Zymogram and sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the presence of a protein band corresponding to the cellulolytic activity in the molecular weight (MW) range of approximately 28,000. Protein bands in the same range were detected in five Clavibacter michiganensis subsp. sepedonicus strains. Studies on crude enzyme extracts of Clavibacter michiganensis subsp. sepedonicus strain N-1-1 revealed that p-nitrophenyl beta-D-cellobioside (pNPC) was hydrolyzed, with optimal activity at 37 degrees C and pH 7.0.

  2. Study of cellulolytic soil fungi and two nova species and new medium

    Institute of Scientific and Technical Information of China (English)

    KHALID Mahmood; YANG Wei-jun; KISHWAR Nazir; RAJPUT Zahid Iqbal; ARIJO Abdullah G.

    2006-01-01

    This study is aimed at identifying and determining the percentage of occurrence frequency of cellulose decomposing soil fungi. The soil samples were inoculated into culture plates prepared in Sabouraud medium under sterilized conditions and incubated at 30 ℃ for 4 to 7 d. The identified fungal species were incubated in self-designed cellulose medium for testing their cellulolytic ability. Forty-two species, including 2 nova species, representing sixteen genera showed growth and sporulation in the cellulose medium. Most of the isolated species were from genus Aspergillus and Penicillium. Aspergillus niger and Mucor hiemalis showed highest occurrence frequency (45% and 36% respectively), as these species were collected from about 80% of soil samples. Being agar free and cheaper, the new fungal medium designed showed results equivalent to Sabouraud medium.

  3. Cellulolytic and xylanolytic potential of high β-glucosidase-producing Trichoderma from decaying biomass.

    Science.gov (United States)

    Okeke, Benedict C

    2014-10-01

    Availability, cost, and efficiency of microbial enzymes for lignocellulose bioconversion are central to sustainable biomass ethanol technology. Fungi enriched from decaying biomass and surface soil mixture displayed an array of strong cellulolytic and xylanolytic activities. Strains SG2 and SG4 produced a promising array of cellulolytic and xylanolytic enzymes including β-glucosidase, usually low in cultures of Trichoderma species. Nucleotide sequence analysis of internal transcribed spacer 2 (ITS2) region of rRNA gene revealed that strains SG2 and SG4 are closely related to Trichoderma inhamatum, Trichoderma piluliferum, and Trichoderma aureoviride. Trichoderma sp. SG2 crude culture supernatant correspondingly displayed as much as 9.84 ± 1.12, 48.02 ± 2.53, and 30.10 ± 1.11 units mL(-1) of cellulase, xylanase, and β-glucosidase in 30 min assay. Ten times dilution of culture supernatant of strain SG2 revealed that total activities were about 5.34, 8.45, and 2.05 orders of magnitude higher than observed in crude culture filtrate for cellulase, xylanase, and β-glucosidase, respectively, indicating that more enzymes are present to contact with substrates in biomass saccharification. In parallel experiments, Trichoderma species SG2 and SG4 produced more β-glucosidase than the industrial strain Trichoderma reesei RUT-C30. Results indicate that strains SG2 and SG4 have potential for low cost in-house production of primary lignocellulose-hydrolyzing enzymes for production of biomass saccharides and biofuel in the field.

  4. Microbial Consortium with High Cellulolytic Activity (MCHCA for enhanced biogas production.

    Directory of Open Access Journals (Sweden)

    Krzysztof ePoszytek

    2016-03-01

    Full Text Available The use of lignocellulosic biomass as a substrate in agricultural biogas plants is very popular and yields good results. However, the efficiency of anaerobic digestion, and thus biogas production, is not always satisfactory due to the slow or incomplete degradation (hydrolysis of plant matter. To enhance the solubilization of the lignocellulosic biomass various physical, chemical and biological pretreatment methods are used.The aim of this study was to select and characterize cellulose-degrading bacteria, and to construct a microbial consortium, dedicated for degradation of maize silage and enhancing biogas production from this substrate.Over one hundred strains of cellulose-degrading bacteria were isolated from: sewage sludge, hydrolyzer from an agricultural biogas plant, cattle slurry and manure. After physiological characterization of the isolates, sixteen strains (representatives of Bacillus, Providencia and Ochrobactrum genera were chosen for the construction of a Microbial Consortium with High Cellulolytic Activity, called MCHCA. The selected strains had a high endoglucanase activity (exceeding 0.21 IU/mL CMCase activity and a wide range of tolerance to various physical and chemical conditions. Lab-scale simulation of biogas production using the selected strains for degradation of maize silage was carried out in a two-bioreactor system, similar to those used in agricultural biogas plants.The obtained results showed that the constructed MCHCA consortium is capable of efficient hydrolysis of maize silage, and increases biogas production by even 38%, depending on the inoculum used for methane fermentation. The results in this work indicate that the mesophilic Microbial Consortium with High Cellulolytic Activity has a great potential for application on industrial scale in agricultural biogas plants.

  5. Genes regulated by AoXlnR, the xylanolytic and cellulolytic transcriptional regulator, in Aspergillus oryzae.

    Science.gov (United States)

    Noguchi, Yuji; Sano, Motoaki; Kanamaru, Kyoko; Ko, Taro; Takeuchi, Michio; Kato, Masashi; Kobayashi, Tetsuo

    2009-11-01

    XlnR is a Zn(II)2Cys6 transcriptional activator of xylanolytic and cellulolytic genes in Aspergillus. Overexpression of the aoxlnR gene in Aspergillus oryzae (A. oryzae xlnR gene) resulted in elevated xylanolytic and cellulolytic activities in the culture supernatant, in which nearly 40 secreted proteins were detected by two-dimensional electrophoresis. DNA microarray analysis to identify the transcriptional targets of AoXlnR led to the identification of 75 genes that showed more than fivefold increase in their expression in the AoXlnR overproducer than in the disruptant. Of these, 32 genes were predicted to encode a glycoside hydrolase, highlighting the biotechnological importance of AoXlnR in biomass degradation. The 75 genes included the genes previously identified as AoXlnR targets (xynF1, xynF3, xynG2, xylA, celA, celB, celC, and celD). Thirty-six genes were predicted to be extracellular, which was consistent with the number of proteins secreted, and 61 genes possessed putative XlnR-binding sites (5'-GGCTAA-3', 5'-GGCTAG-3', and 5'-GGCTGA-3') in their promoter regions. Functional annotation of the genes revealed that AoXlnR regulated the expression of hydrolytic genes for degradation of beta-1,4-xylan, arabinoxylan, cellulose, and xyloglucan and of catabolic genes for the conversion of D-xylose to xylulose-5-phosphate. In addition, genes encoding glucose-6-phosphate 1-dehydrogenase and L-arabinitol-4- dehydrogenase involved in D-glucose and L-arabinose catabolism also appeared to be targets of AoXlnR.

  6. Alkaloids from the Mangrove-Derived Actinomycete Jishengella endophytica 161111

    Directory of Open Access Journals (Sweden)

    Pei Wang

    2014-01-01

    Full Text Available A new alkaloid, 2-(furan-2-yl-6-(2S,3S,4-trihydroxybutylpyrazine (1, along with 12 known compounds, 2-(furan-2-yl-5-(2S,3S,4-trihydroxybutylpyrazine (2, (S-4-isobutyl-3-oxo-3,4-dihydro-1H-pyrrolo[2,1-c][1,4]oxazine-6-carbaldehyde (3, (S-4-isopropyl-3-oxo-3,4-dihydro-1H-pyrrolo[2,1-c][1,4]oxazine-6-carbaldehyde (4, (4S-4-(2-methylbutyl-3-oxo-3,4-dihydro-1H-pyrrolo[2,1-c][1,4]oxazine-6-carbaldehyde (5, (S-4-benzyl-3-oxo-3,4-dihydro-1H-pyrrolo[2,1-c][1,4]oxazine-6-carbaldehyde (6, flazin (7, perlolyrine (8, 1-hydroxy-β-carboline (9, lumichrome (10, 1H-indole-3-carboxaldehyde (11, 2-hydroxy-1-(1H-indol-3-ylethanone (12, and 5-(methoxymethyl-1H-pyrrole-2-carbaldehyde (13, were isolated and identified from the fermentation broth of an endophytic actinomycetes, Jishengella endophytica 161111. The new structure 1 and the absolute configurations of 2–6 were determined by spectroscopic methods, J-based configuration analysis (JBCA method, lactone sector rule, and electronic circular dichroism (ECD calculations. Compounds 8–11 were active against the influenza A virus subtype H1N1 with IC50 and selectivity index (SI values of 38.3(±1.2/25.0(±3.6/ 39.7(±5.6/45.9(±2.1 μg/mL and 3.0/16.1/3.1/11.4, respectively. The IC50 and SI values of positive control, ribavirin, were 23.1(±1.7 μg/mL and 32.2, respectively. The results showed that compound 9 could be a promising new hit for anti-H1N1 drugs. The absolute configurations of 2–5, 13C nuclear magnetic resonance (NMR data and the specific rotations of 3–6 were also reported here for the first time.

  7. Isolation and characterization of marine-derived actinomycetes with cytotoxic activity from the Red Sea coast

    Directory of Open Access Journals (Sweden)

    Mohamed Saleh Abdelfattah

    2016-08-01

    Conclusions: The results of the present study reveal that marine sediments of the Red Sea are a potent source of novel species of actinomycetes. The isolates may be useful in discovery of novel bioactive compounds and an important step in the development of microbial natural product research.

  8. Biodiversity, Anti-Trypanosomal Activity Screening, and Metabolomic Profiling of Actinomycetes Isolated from Mediterranean Sponges.

    Directory of Open Access Journals (Sweden)

    Cheng Cheng

    Full Text Available Marine sponge-associated actinomycetes are considered as promising sources for the discovery of novel biologically active compounds. In the present study, a total of 64 actinomycetes were isolated from 12 different marine sponge species that had been collected offshore the islands of Milos and Crete, Greece, eastern Mediterranean. The isolates were affiliated to 23 genera representing 8 different suborders based on nearly full length 16S rRNA gene sequencing. Four putatively novel species belonging to genera Geodermatophilus, Microlunatus, Rhodococcus and Actinomycetospora were identified based on a 16S rRNA gene sequence similarity of < 98.5% to currently described strains. Eight actinomycete isolates showed bioactivities against Trypanosma brucei brucei TC221 with half maximal inhibitory concentration (IC50 values <20 μg/mL. Thirty four isolates from the Milos collection and 12 isolates from the Crete collection were subjected to metabolomic analysis using high resolution LC-MS and NMR for dereplication purposes. Two isolates belonging to the genera Streptomyces (SBT348 and Micromonospora (SBT687 were prioritized based on their distinct chemistry profiles as well as their anti-trypanosomal activities. These findings demonstrated the feasibility and efficacy of utilizing metabolomics tools to prioritize chemically unique strains from microorganism collections and further highlight sponges as rich source for novel and bioactive actinomycetes.

  9. An N-acyl homolog of mycothiol is produced in marine actinomycetes.

    Science.gov (United States)

    Newton, Gerald L; Jensen, Paul R; Macmillan, John B; Fenical, William; Fahey, Robert C

    2008-11-01

    Marine actinomycetes have generated much recent interest as a potentially valuable source of novel antibiotics. Like terrestrial actinomycetes the marine actinomycetes are shown here to produce mycothiol as their protective thiol. However, a novel thiol, U25, was produced by MAR2 strain CNQ703 upon progression into stationary phase when secondary metabolite production occurred and became the dominant thiol. MSH and U25 were maintained in a reduced state during early stationary phase, but become significantly oxidized after 10 days in culture. Isolation and structural analysis of the monobromobimane derivative identified U25 as a homolog of mycothiol in which the acetyl group attached to the nitrogen of cysteine is replaced by a propionyl residue. This N-propionyl-desacetyl-mycothiol was present in 13 of the 17 strains of marine actinomycetes examined, including five strains of Salinispora and representatives of the MAR2, MAR3, MAR4 and MAR6 groups. Mycothiol and its precursor, the pseudodisaccharide 1-O-(2-amino-2-deoxy-alpha-D-glucopyranosyl)-D-myo-inositol, were found in all strains. High levels of mycothiol S-conjugate amidase activity, a key enzyme in mycothiol-dependent detoxification, were found in most strains. The results demonstrate that major thiol/disulfide changes accompany secondary metabolite production and suggest that mycothiol-dependent detoxification is important at this developmental stage.

  10. Bioprospecting of Novel and Bioactive Compounds from Marine Actinomycetes Isolated from South China Sea Sediments.

    Science.gov (United States)

    Yang, Na; Song, Fuhang

    2017-09-16

    Marine actinomycetes are less investigated compared to terrestrial strains as potential sources of natural products. To date, few investigations have been performed on culturable actinomycetes associated with South China Sea sediments. In the present study, twenty-eight actinomycetes were recovered from South China Sea sediments after dereplication by traditional culture-dependent method. The 16S rRNA gene sequences analyses revealed that these strains related to five families and seven genera. Twelve representative strains possessed at least one of the biosynthetic genes coding for polyketide synthase I, II, and nonribosomal peptide synthetase. Four strains had anti-Mycobacterium phlei activities and five strains had activities against methicillin-resistant Staphylococcus aureus. 10 L-scale fermentation of strains Salinispora sp. NHF45, Nocardiopsis sp. NHF48, and Streptomyces sp. NHF86 were carried out for novel and bioactive compounds discovery. Finally, we obtained a novel α-pyrone compound from marine Nocardiopsis sp. NHF48, an analogue of paulomenol from marine Streptomyces sp. NHF86 and a new source of rifamycin B, produced by Salinispora sp. NHF45. The present study concluded that marine actinomycetes, which we isolated from South China Sea sediments, will be a suitable source for the development of novel and bioactive compounds.

  11. [Bioactivity of endophytic actinomycetes from medicinal plants and secondary metabolites from strain D62].

    Science.gov (United States)

    Liu, Ning; Zhang, Hui; Zheng, Wen; Huang, Ying; Wang, Hai-Bin

    2007-10-01

    It is believed that genetic recombination of the endophytes with the hosts that occurred in evolutionary time could result in some endophytes producing certain phytochemical originally characteristic of the host. Based on this widely accepted hypothesis, there have been increasing research efforts focused on screening for novel natural products from endophytes. In this study, antimicrobial and antitumor activities of 165 actinomycetes isolated from medicinal plants collected from Xishuangbanna were tested by agar diffusion method and WST-8 assay respectively. The results showed that over 42% of the isolates exhibited antagonism against pathogenic strains, and 54.5% displayed excellent inhibition against mouse melanoma cell line B16 or/and human alveolar epithelial cell line A549. These results are superior to those of soil actinomycetes, indicating tremendous potential of endophytic of actinomycetes for exploration. Six compounds that had both antimicrobial and antitumor activities were separated and purified from isolate Streptomyces sp. D62 by resin adsorption, silica-gel column and sephadex chromatography, etc. On the basis of spectral analyses, they were identified as antimycin A4a (1), antimycin A7a (2), antimycin A2a (3), antimycin A1a (4), 10-hydroxy-10-methyl-dodec-2-en-1,4-olide (5) and 6-(2-(4-aminophenyl)-2-oxoethyl)-3,5-dimethyl-tetrahydropyran-2-one(6), with the last one defined as a novel compound. Based on all these results, it is convinced that endophytic actinomycetes are a promising resource for bioactive natural product discovery.

  12. Actinomycetes inhibit filamentous fungi from the cuticle of Acromyrmex leafcutter ants.

    Science.gov (United States)

    Dângelo, Rômulo Augusto Cotta; de Souza, Danival José; Mendes, Thais Demarchi; Couceiro, Joel da Cruz; Lucia, Terezinha Maria Castro Della

    2016-03-01

    Actinomycetes bacteria associated with leafcutter ants produce secondary metabolites with antimicrobial properties against Escovopsis, a fungus specialized in attacking the gardens of fungus-growing ants, which denies the ants their food source. Because previous studies have used fungi isolated from fungus gardens but not from ant integument, the aims of the present study were to isolate actinomycetes associated with the cuticle of the Acromyrmex spp. and to quantify their inhibition abilities against the filamentous fungal species carried by these ants. The results demonstrated that actinomycetes had varied strain-dependent effects on several filamentous fungal species in addition to antagonistic activity against Escovopsis. The strain isolated from Acromyrmex balzani was identified as a Streptomyces species, whereas the remaining isolates were identified as different strains belonging to the genus Pseudonocardia. These findings corroborate the hypothesis that actinomycetes do not act specifically against Escovopsis mycoparasites and may have the ability to inhibit other species of pathogenic fungi. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Genome Sequence of Streptomyces viridosporus Strain T7A ATCC 39115, a Lignin-Degrading Actinomycete

    Science.gov (United States)

    Davis, Jennifer R.; Goodwin, Lynne; Teshima, Hazuki; Detter, Chris; Tapia, Roxanne; Han, Cliff; Huntemann, Marcel; Wei, Chia-Lin; Han, James; Chen, Amy; Kyrpides, Nikos; Mavrommatis, Kostas; Szeto, Ernest; Markowitz, Victor; Ivanova, Natalia; Mikhailova, Natalia; Ovchinnikova, Galina; Pagani, Ioanna; Pati, Amrita; Woyke, Tanja; Pitluck, Sam; Peters, Lin; Nolan, Matt; Land, Miriam

    2013-01-01

    We announce the availability of the genome sequence of Streptomyces viridosporus strain T7A ATCC 39115, a plant biomass-degrading actinomycete. This bacterium is of special interest because of its capacity to degrade lignin, an underutilized component of plants in the context of bioenergy. It has a full complement of genes for plant biomass catabolism. PMID:23833133

  14. Effect of gamma radiation on the survival of fungal and actinomycetal florae contaminating medicinal plants

    Energy Technology Data Exchange (ETDEWEB)

    Aziz, N.H.; El-Fouly, M.Z.; Moussa, L.A.A. [National Center for Radiation Research and Technology, Cairo (Egypt); Abu-Shady, M.R. [Ain Shams Univ., Cairo (Egypt). Faculty of Science

    1997-01-01

    This study evaluates the effect of gamma radiation on the viability of fungi and actinomycetes that contaminate medicinal plants. The relationship between the total lipids of some fungi and actinomycetes and their sensitivity to gamma radiation is also investigated. The data reveal that the viable counts of these florae decrease approximately exponentially with the radiation dose, the effective dose for the elimination of these microorganisms being about 5 kGy for all the medicinal plants under study. Response of pure cultures of fungi and actinomycetes isolated from medicinal plants to increasing absorbed doses of gamma radiation indicate that an increase in radioresistance is in the following order: Streptomyces rimosus, Fusarium solani, Nocardia kuroishii. F. oxysporum, A. fumigatus, A. flavus, A. parasiticus and A. ochraceus. The total lipid contents of molds and actinomycetes have been reported to be increased by increasing the radio-resistance of microorganisms, and hence there is a relationship between the radio-sensitivity of microorganisms and the total lipid mass of flora mycelia. (Author).

  15. Genome Sequence of Streptomyces viridosporus Strain T7A ATCC 39115, a Lignin-Degrading Actinomycete

    Energy Technology Data Exchange (ETDEWEB)

    Davis, Jennifer R. [Brown University; Goodwin, Lynne A. [Los Alamos National Laboratory (LANL); Teshima, Hazuki [Los Alamos National Laboratory (LANL); Detter, J. Chris [U.S. Department of Energy, Joint Genome Institute; Tapia, Roxanne [Los Alamos National Laboratory (LANL); Han, Cliff [Los Alamos National Laboratory (LANL); Huntemann, Marcel [U.S. Department of Energy, Joint Genome Institute; Wei, Chia-Lin [Los Alamos National Laboratory (LANL); Han, James [U.S. Department of Energy, Joint Genome Institute; Chen, Amy [U.S. Department of Energy, Joint Genome Institute; Kyrpides, Nikos C [U.S. Department of Energy, Joint Genome Institute; Mavromatis, K [U.S. Department of Energy, Joint Genome Institute; Szeto, Ernest [U.S. Department of Energy, Joint Genome Institute; Markowitz, Victor [U.S. Department of Energy, Joint Genome Institute; Ivanova, N [U.S. Department of Energy, Joint Genome Institute; Mikhailova, Natalia [U.S. Department of Energy, Joint Genome Institute; Ovchinnikova, Galina [U.S. Department of Energy, Joint Genome Institute; Pagani, Ioanna [U.S. Department of Energy, Joint Genome Institute; Pati, Amrita [U.S. Department of Energy, Joint Genome Institute; Woyke, Tanja [U.S. Department of Energy, Joint Genome Institute; Pitluck, Sam [U.S. Department of Energy, Joint Genome Institute; Peters, Lin [U.S. Department of Energy, Joint Genome Institute; Nolan, Matt [U.S. Department of Energy, Joint Genome Institute; Land, Miriam L [ORNL; Sello, Jason K. [Brown University

    2013-01-01

    We announce the availability of the genome sequence of Streptomyces viridosporus strain T7A ATCC 39115, a plant biomass- degrading actinomycete. This bacterium is of special interest because of its capacity to degrade lignin, an underutilized compo- nent of plants in the context of bioenergy. It has a full complement of genes for plant biomass catabolism.

  16. Comparative genome-scale metabolic modeling of actinomycetes : The topology of essential core metabolism

    NARCIS (Netherlands)

    Alam, Mohammad Tauqeer; Medema, Marnix H.; Takano, Eriko; Breitling, Rainer; Gojobori, Takashi

    2011-01-01

    Actinomycetes are highly important bacteria. On one hand, some of them cause severe human and plant diseases, on the other hand, many species are known for their ability to produce antibiotics. Here we report the results of a comparative analysis of genome-scale metabolic models of 37 species of act

  17. Comparative genome-scale metabolic modeling of actinomycetes: the topology of essential core metabolism.

    NARCIS (Netherlands)

    Alam, M.T.; Medema, M.H.; Takano, E.; Breitling, R.

    2011-01-01

    Actinomycetes are highly important bacteria. On one hand, some of them cause severe human and plant diseases, on the other hand, many species are known for their ability to produce antibiotics. Here we report the results of a comparative analysis of genome-scale metabolic models of 37 species of act

  18. Characterization of cytotoxic compound from marine sediment derived actinomycete Streptomyces avidinii strain SU4

    Directory of Open Access Journals (Sweden)

    S Sudha

    2012-10-01

    Conclusion: This study clearly proves that the marine sediment derived actinomycetes with bioactive metabolites can be expected to provide high quality biological material for high throughout biochemical and anticancer screening programs. These results help us to conclude that the potential of using metabolic engineering and post genomic approaches to isolate more bioactive compounds and make their possible commercial application is not far off.

  19. Comparative genome-scale metabolic modeling of actinomycetes: the topology of essential core metabolism.

    NARCIS (Netherlands)

    Alam, M.T.; Medema, M.H.; Takano, E.; Breitling, R.

    2011-01-01

    Actinomycetes are highly important bacteria. On one hand, some of them cause severe human and plant diseases, on the other hand, many species are known for their ability to produce antibiotics. Here we report the results of a comparative analysis of genome-scale metabolic models of 37 species of

  20. Comparative genome-scale metabolic modeling of actinomycetes : The topology of essential core metabolism

    NARCIS (Netherlands)

    Alam, Mohammad Tauqeer; Medema, Marnix H.; Takano, Eriko; Breitling, Rainer; Gojobori, Takashi

    2011-01-01

    Actinomycetes are highly important bacteria. On one hand, some of them cause severe human and plant diseases, on the other hand, many species are known for their ability to produce antibiotics. Here we report the results of a comparative analysis of genome-scale metabolic models of 37 species of

  1. Isolation and identification of actinomycetes from a compost-amended soils biocontrol agents

    Energy Technology Data Exchange (ETDEWEB)

    Garcia de la Fuente, R.; Cuesta, G.; Fornes, F.; Abad, M.

    2009-07-01

    Compost capability to suppress soil-borne plant pathogens has become an interesting subject as a strategy for reducing the adverse effects of massive fungicides application in the environmental. In this context, actinomycetes have received considerable attention as biocontrol agents, particularly Streptomyces species. (Author)

  2. Biodiversity of Actinomycetes associated with Caribbean sponges and their potential for natural product discovery.

    Science.gov (United States)

    Vicente, Jan; Stewart, Allison; Song, Bongkeun; Hill, Russell T; Wright, Jeffrey L

    2013-08-01

    Marine actinomycetes provide a rich source of structurally unique and bioactive secondary metabolites. Numerous genera of marine actinomycetes have been isolated from marine sediments as well as several sponge species. In this study, 16 different species of Caribbean sponges were collected from four different locations in the coastal waters off Puerto Rico in order to examine diversity and bioactive metabolite production of marine actinomycetes in Caribbean sponges. Sediments were also collected from each location, in order to compare actinomycete communities between these two types of samples. A total of 180 actinomycetes were isolated and identified based on 16S rRNA gene analysis. Phylogenetic analysis revealed the presence of at least 14 new phylotypes belonging to the genera Micromonospora, Verruscosispora, Streptomyces, Salinospora, Solwaraspora, Microbacterium and Cellulosimicrobium. Seventy-eight of the isolates (19 from sediments and 59 from sponges) shared 100 % sequence identity with Micromonospora sp. R1. Despite having identical 16S rRNA sequences, the bioactivity of extracts and subsequent fractions generated from the fermentation of both sponge- and sediment-derived isolates identical to Micromonospora sp. R1 varied greatly, with a marked increase in antibiotic metabolite production in those isolates derived from sponges. These results indicate that the chemical profiles of isolates with high 16S rRNA sequence homology to known strains can be diverse and dependent on the source of isolation. In addition, seven previously reported dihydroquinones produced by five different Streptomyces strains have been purified and characterized from one Streptomyces sp. strain isolated in this study from the Caribbean sponge Agelas sceptrum.

  3. A mixed community of actinomycetes produce multiple antibiotics for the fungus farming ant Acromyrmex octospinosus

    Directory of Open Access Journals (Sweden)

    Barke Jörg

    2010-08-01

    Full Text Available Abstract Background Attine ants live in an intensely studied tripartite mutualism with the fungus Leucoagaricus gongylophorus, which provides food to the ants, and with antibiotic-producing actinomycete bacteria. One hypothesis suggests that bacteria from the genus Pseudonocardia are the sole, co-evolved mutualists of attine ants and are transmitted vertically by the queens. A recent study identified a Pseudonocardia-produced antifungal, named dentigerumycin, associated with the lower attine Apterostigma dentigerum consistent with the idea that co-evolved Pseudonocardia make novel antibiotics. An alternative possibility is that attine ants sample actinomycete bacteria from the soil, selecting and maintaining those species that make useful antibiotics. Consistent with this idea, a Streptomyces species associated with the higher attine Acromyrmex octospinosus was recently shown to produce the well-known antifungal candicidin. Candicidin production is widespread in environmental isolates of Streptomyces, so this could either be an environmental contaminant or evidence of recruitment of useful actinomycetes from the environment. It should be noted that the two possibilities for actinomycete acquisition are not necessarily mutually exclusive. Results In order to test these possibilities we isolated bacteria from a geographically distinct population of A. octospinosus and identified a candicidin-producing Streptomyces species, which suggests that they are common mutualists of attine ants, most probably recruited from the environment. We also identified a Pseudonocardia species in the same ant colony that produces an unusual polyene antifungal, providing evidence for co-evolution of Pseudonocardia with A. octospinosus. Conclusions Our results show that a combination of co-evolution and environmental sampling results in the diversity of actinomycete symbionts and antibiotics associated with attine ants.

  4. Molecular, chemical and biological screening of soil actinomycete isolates in seeking bioactive peptide metabolites

    Directory of Open Access Journals (Sweden)

    Javad Hamedi

    2015-10-01

    Full Text Available Background and Objective: Due to the evolution of multidrug-resistant strains, screening of natural resources, especially actinomycetes, for new therapeutic agents discovery has become the interests of researchers. In this study, molecular, chemical and biological screening of soil actinomycetes was carried out in order to search for peptide-producing actinomycetes.Materials and Methods: 60 actinomycetes were isolated from soils of Iran. The isolates were subjected to molecular screening for detection NRPS (non-ribosomal peptide synthetases gene. Phylogenic identification of NRPS containing isolates was performed. Chemical screening of the crude extracts was performed using chlorine o-dianisidine as peptide detector reagent and bioactivity of peptide producing strains was determined by antimicrobial bioassay. High pressure liquid chromatography- mass spectrometry (HPLC-MS with UV-visible spectroscopy was performed for detection of the metabolite diversity in selected strain.Results: Amplified NRPS adenylation gene (700 bp was detected among 30 strains. Phylogenic identification of these isolates showed presence of rare actinomycetes genera among the isolates and 10 out of 30 strains were subjected to chemical screening. Nocardia sp. UTMC 751 showed antimicrobial activity against bacterial and fungal test pathogens. HPLC-MSand UV-visible spectroscopy results from the crude extract showed that this strain has probably the ability to produce new metabolites.Conclusion: By application of a combined approach, including molecular, chemical and bioactivity analysis, a promising strain of Nocardia sp. UTMC 751 was obtained. This strain had significant activity against Staphylococcus aureus and Pseudomonas aeruginosa. Strain Nocardia sp. UTMC 751 produce five unknown and most probably new metabolites with molecular weights of 274.2, 390.3, 415.3, 598.4 and 772.5. This strain had showed 99% similarity to Nocardia ignorata DSM 44496 T.

  5. Isolation and phylogenetic assignation of actinomycetes in the marine sediments from the Arctic Ocean

    Institute of Scientific and Technical Information of China (English)

    YU Yong; LI Huirong; ZENG Yinxin; CHEN Bo

    2005-01-01

    Actinomycetes in five marine sediments collected from the Arctic Ocean at depths of 43 to 3 050 m were cultivated using a variety of media. A total of 61 actinomycete colonies with substrate mycelia only were observed, and no colonies with aerial mycelia were observed under aerobic conditions at 15 ℃. From these colonies, 28 were selected to represent different morphological types.Denaturing gradient gel electrophoresis (DGGE) was used to check the purity of isolates and select representatives for subsequent sequencing. Phylogentic analyses based on nearly full-length 16S ribosomal RNA gene (rDNA) sequences indicated that the actinomycetes isolated were accommodated within genus Rhodococcus of family Nocardiaceae, genus Dietzia of family Dietziaceae,genera Janibacter and Terrabacter of family Instrasporangiaceae and genera Kocuria and Arthrobacter of family Micrococcaceae. One of the strains (P27-24) from the deep-sea sediment at depth of 3 050 m was found to be identical in 16S rDNA sequence(1474/1474)with the radiation-resistant Kocuria rosea ATCC 187T isolated from air. More than halfofthe isolates showed the similarities ranging from 99.5% to 99.9% in 16S rDNA sequence to dibenzofran-degrading, butyl 2-ethylhexanoate-hydrolysising and nitrile-metabolizing actinomycetes. All the strains isolated were psychrotolerant bacteria and grew better on the media prepared with natural seawater than on the media prepared with deionized water. Three of them (Dietzia sp. P27-10, Rhodococcus sp. S11-3 and Rhodococcus sp.P11-5)had an obligate growth requirement for salt, confirming that these strains are indigenous marine actinomycetes.

  6. Thermophilic and cellulolytic consortium isolated from composting plants improves anaerobic digestion of cellulosic biomass: Toward a microbial resource management approach.

    Science.gov (United States)

    Kinet, R; Destain, J; Hiligsmann, S; Thonart, P; Delhalle, L; Taminiau, B; Daube, G; Delvigne, F

    2015-01-01

    A cellulolytic consortium was isolated from a composting plant in order to boost the initial hydrolysis step encountered in anaerobic digestion. Improvement of the cellulose degradation, as well as biogas production, was observed for the cultures inoculated with the exogenous consortium. Metagenomics analyses pointed out a weak richness (related to the number of OTUs) of the exogenous consortium induced by the selective pressure (cellulose as sole carbon source) met during the initial isolation steps. Main microbial strains determined were strictly anaerobic and belong to the Clostridia class. During cellulose anaerobic degradation, pH drop induced a strong modification of the microbial population. Despite the fact that richness and evenness were very weak, the exogenous consortium was able to adapt and to maintain the cellulolytic degradation potential. This important result point out the fact that simplified microbial communities could be used in order to increase the robustness of mixed cultures involved in environmental biotechnology. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. Ultrastructural studies of the termite (Odontotermes obesus) gut microflora and its cellulolytic properties.

    Science.gov (United States)

    Paul, J; Saxena, S; Varma, A

    1993-01-01

    The major gut microflora colonizing the hind gut of a higher termite,Odontotermes obesus, included morphologically diverse bacteria, both coccoid and rod-shaped, along with spirochaetes, pseudomonads and actinomycetes. Flagellated protozoa were totally absent. When the gut extract was inoculated on plates containing carboxymethyl cellulose or cellobiose, higher numbers of bacteria grew than on plates without cellulosic sources. The gut homogenate exhibited strong hydrolytic activity when carboxymethyl cellulose,p-nitrophenyl-β-D-glucoside or xylan were used as substrate, indicating the role of gut microbiota in the process of cellulose and hemicellulose digestion. Activities were highest in the hind gut, and the paunch was probably the major site of polysaccharide digestion in this higher termite.In vitro cultivation of some of the isolates revealed both cellulase and xylanase activities. To our knowledge, this is the first report on ultrastructural studies of the higher termiteOdontotermes obesus.

  8. Evolution of High Cellulolytic Activity in Symbiotic Streptomyces through Selection of Expanded Gene Content and Coordinated Gene Expression.

    Directory of Open Access Journals (Sweden)

    Adam J Book

    2016-06-01

    Full Text Available The evolution of cellulose degradation was a defining event in the history of life. Without efficient decomposition and recycling, dead plant biomass would quickly accumulate and become inaccessible to terrestrial food webs and the global carbon cycle. On land, the primary drivers of plant biomass deconstruction are fungi and bacteria in the soil or associated with herbivorous eukaryotes. While the ecological importance of plant-decomposing microbes is well established, little is known about the distribution or evolution of cellulolytic activity in any bacterial genus. Here we show that in Streptomyces, a genus of Actinobacteria abundant in soil and symbiotic niches, the ability to rapidly degrade cellulose is largely restricted to two clades of host-associated strains and is not a conserved characteristic of the Streptomyces genus or host-associated strains. Our comparative genomics identify that while plant biomass degrading genes (CAZy are widespread in Streptomyces, key enzyme families are enriched in highly cellulolytic strains. Transcriptomic analyses demonstrate that cellulolytic strains express a suite of multi-domain CAZy enzymes that are coregulated by the CebR transcriptional regulator. Using targeted gene deletions, we verify the importance of a highly expressed cellulase (GH6 family cellobiohydrolase and the CebR transcriptional repressor to the cellulolytic phenotype. Evolutionary analyses identify complex genomic modifications that drive plant biomass deconstruction in Streptomyces, including acquisition and selective retention of CAZy genes and transcriptional regulators. Our results suggest that host-associated niches have selected some symbiotic Streptomyces for increased cellulose degrading activity and that symbiotic bacteria are a rich biochemical and enzymatic resource for biotechnology.

  9. Evolution of High Cellulolytic Activity in Symbiotic Streptomyces through Selection of Expanded Gene Content and Coordinated Gene Expression.

    Science.gov (United States)

    Book, Adam J; Lewin, Gina R; McDonald, Bradon R; Takasuka, Taichi E; Wendt-Pienkowski, Evelyn; Doering, Drew T; Suh, Steven; Raffa, Kenneth F; Fox, Brian G; Currie, Cameron R

    2016-06-01

    The evolution of cellulose degradation was a defining event in the history of life. Without efficient decomposition and recycling, dead plant biomass would quickly accumulate and become inaccessible to terrestrial food webs and the global carbon cycle. On land, the primary drivers of plant biomass deconstruction are fungi and bacteria in the soil or associated with herbivorous eukaryotes. While the ecological importance of plant-decomposing microbes is well established, little is known about the distribution or evolution of cellulolytic activity in any bacterial genus. Here we show that in Streptomyces, a genus of Actinobacteria abundant in soil and symbiotic niches, the ability to rapidly degrade cellulose is largely restricted to two clades of host-associated strains and is not a conserved characteristic of the Streptomyces genus or host-associated strains. Our comparative genomics identify that while plant biomass degrading genes (CAZy) are widespread in Streptomyces, key enzyme families are enriched in highly cellulolytic strains. Transcriptomic analyses demonstrate that cellulolytic strains express a suite of multi-domain CAZy enzymes that are coregulated by the CebR transcriptional regulator. Using targeted gene deletions, we verify the importance of a highly expressed cellulase (GH6 family cellobiohydrolase) and the CebR transcriptional repressor to the cellulolytic phenotype. Evolutionary analyses identify complex genomic modifications that drive plant biomass deconstruction in Streptomyces, including acquisition and selective retention of CAZy genes and transcriptional regulators. Our results suggest that host-associated niches have selected some symbiotic Streptomyces for increased cellulose degrading activity and that symbiotic bacteria are a rich biochemical and enzymatic resource for biotechnology.

  10. Pyrosequencing reveals high-temperature cellulolytic microbial consortia in Great Boiling Spring after in situ lignocellulose enrichment.

    Directory of Open Access Journals (Sweden)

    Joseph P Peacock

    Full Text Available To characterize high-temperature cellulolytic microbial communities, two lignocellulosic substrates, ammonia fiber-explosion-treated corn stover and aspen shavings, were incubated at average temperatures of 77 and 85°C in the sediment and water column of Great Boiling Spring, Nevada. Comparison of 109,941 quality-filtered 16S rRNA gene pyrosequences (pyrotags from eight enrichments to 37,057 quality-filtered pyrotags from corresponding natural samples revealed distinct enriched communities dominated by phylotypes related to cellulolytic and hemicellulolytic Thermotoga and Dictyoglomus, cellulolytic and sugar-fermenting Desulfurococcales, and sugar-fermenting and hydrogenotrophic Archaeoglobales. Minor enriched populations included close relatives of hydrogenotrophic Thermodesulfobacteria, the candidate bacterial phylum OP9, and candidate archaeal groups C2 and DHVE3. Enrichment temperature was the major factor influencing community composition, with a negative correlation between temperature and richness, followed by lignocellulosic substrate composition. This study establishes the importance of these groups in the natural degradation of lignocellulose at high temperatures and suggests that a substantial portion of the diversity of thermophiles contributing to consortial cellulolysis may be contained within lineages that have representatives in pure culture.

  11. Prospection and Evaluation of (Hemi) Cellulolytic Enzymes Using Untreated and Pretreated Biomasses in Two Argentinean Native Termites

    Science.gov (United States)

    Ben Guerrero, Emiliano; Arneodo, Joel; Bombarda Campanha, Raquel; Abrão de Oliveira, Patrícia; Veneziano Labate, Mônica T.; Regiani Cataldi, Thaís; Campos, Eleonora; Cataldi, Angel; Labate, Carlos A.; Martins Rodrigues, Clenilson; Talia, Paola

    2015-01-01

    Saccharum officinarum bagasse (common name: sugarcane bagasse) and Pennisetum purpureum (also known as Napier grass) are among the most promising feedstocks for bioethanol production in Argentina and Brazil. In this study, both biomasses were assessed before and after acid pretreatment and following hydrolysis with Nasutitermes aquilinus and Cortaritermes fulviceps termite gut digestome. The chemical composition analysis of the biomasses after diluted acid pretreatment showed that the hemicellulose fraction was partially removed. The (hemi) cellulolytic activities were evaluated in bacterial culture supernatants of termite gut homogenates grown in treated and untreated biomasses. In all cases, we detected significantly higher endoglucanase and xylanase activities using pretreated biomasses compared to untreated biomasses, carboxymethylcellulose and xylan. Several protein bands with (hemi) cellulolytic activity were detected in zymograms and two-dimensional gel electrophoresis. Some proteins of these bands or spots were identified as xylanolytic peptides by mass spectrometry. Finally, the diversity of cultured cellulolytic bacterial endosymbionts associated to both Argentinean native termite species was analyzed. This study describes, for the first time, bacterial endosymbionts and endogenous (hemi) cellulases of two Argentinean native termites as well as their potential application in degradation of lignocellulosic biomass for bioethanol production. PMID:26313257

  12. Prospection and Evaluation of (Hemi Cellulolytic Enzymes Using Untreated and Pretreated Biomasses in Two Argentinean Native Termites.

    Directory of Open Access Journals (Sweden)

    Emiliano Ben Guerrero

    Full Text Available Saccharum officinarum bagasse (common name: sugarcane bagasse and Pennisetum purpureum (also known as Napier grass are among the most promising feedstocks for bioethanol production in Argentina and Brazil. In this study, both biomasses were assessed before and after acid pretreatment and following hydrolysis with Nasutitermes aquilinus and Cortaritermes fulviceps termite gut digestome. The chemical composition analysis of the biomasses after diluted acid pretreatment showed that the hemicellulose fraction was partially removed. The (hemi cellulolytic activities were evaluated in bacterial culture supernatants of termite gut homogenates grown in treated and untreated biomasses. In all cases, we detected significantly higher endoglucanase and xylanase activities using pretreated biomasses compared to untreated biomasses, carboxymethylcellulose and xylan. Several protein bands with (hemi cellulolytic activity were detected in zymograms and two-dimensional gel electrophoresis. Some proteins of these bands or spots were identified as xylanolytic peptides by mass spectrometry. Finally, the diversity of cultured cellulolytic bacterial endosymbionts associated to both Argentinean native termite species was analyzed. This study describes, for the first time, bacterial endosymbionts and endogenous (hemi cellulases of two Argentinean native termites as well as their potential application in degradation of lignocellulosic biomass for bioethanol production.

  13. Ras GTPases modulate morphogenesis, sporulation and cellulase gene expression in the cellulolytic fungus Trichoderma reesei.

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    Jiwei Zhang

    Full Text Available BACKGROUND: The model cellulolytic fungus Trichoderma reesei (teleomorph Hypocrea jecorina is capable of responding to environmental cues to compete for nutrients in its natural saprophytic habitat despite its genome encodes fewer degradative enzymes. Efficient signalling pathways in perception and interpretation of environmental signals are indispensable in this process. Ras GTPases represent a kind of critical signal proteins involved in signal transduction and regulation of gene expression. In T. reesei the genome contains two Ras subfamily small GTPases TrRas1 and TrRas2 homologous to Ras1 and Ras2 from S. cerevisiae, but their functions remain unknown. METHODOLOGY/PRINCIPAL FINDINGS: Here, we have investigated the roles of GTPases TrRas1 and TrRas2 during fungal morphogenesis and cellulase gene expression. We show that both TrRas1 and TrRas2 play important roles in some cellular processes such as polarized apical growth, hyphal branch formation, sporulation and cAMP level adjustment, while TrRas1 is more dominant in these processes. Strikingly, we find that TrRas2 is involved in modulation of cellulase gene expression. Deletion of TrRas2 results in considerably decreased transcription of cellulolytic genes upon growth on cellulose. Although the strain carrying a constitutively activated TrRas2(G16V allele exhibits increased cellulase gene transcription, the cbh1 and cbh2 expression in this mutant still strictly depends on cellulose, indicating TrRas2 does not directly mediate the transmission of the cellulose signal. In addition, our data suggest that the effect of TrRas2 on cellulase gene is exerted through regulation of transcript abundance of cellulase transcription factors such as Xyr1, but the influence is independent of cAMP signalling pathway. CONCLUSIONS/SIGNIFICANCE: Together, these findings elucidate the functions for Ras signalling of T. reesei in cellular morphogenesis, especially in cellulase gene expression, which contribute

  14. Specificity of the mutualistic association between actinomycete bacteria and two sympatric species of Acromyrmex leaf-cutting ants

    DEFF Research Database (Denmark)

    Poulsen, M; Cafaro, M; Boomsma, J J

    2005-01-01

    Acromyrmex leaf-cutting ants maintain two highly specialized, vertically transmitted mutualistic ectosymbionts: basidiomycete fungi that are cultivated for food in underground gardens and actinomycete Pseudonocardia bacteria that are reared on the cuticle to produce antibiotics that suppress...... strains. Our finding that individual colonies cultivate a single actinomycete strain is in agreement with predictions from evolutionary theory on host-symbiont conflict over symbiont mixing, but indicates that there may be constraints on the effectiveness of the bacterial symbionts on an evolutionary...

  15. Unique actinomycetes from marine caves and coral reef sediments provide novel PKS and NRPS biosynthetic gene clusters.

    Science.gov (United States)

    Hodges, Tyler W; Slattery, Marc; Olson, Julie B

    2012-06-01

    In the ever-expanding search for novel bioactive molecules and enzymes, marine actinomycetes have proven to be a productive source. While open reef sediment and sponge-associated actinomycetes have been extensively examined, their marine cave counterparts remain unevaluated. Anchialine cave systems in the Bahamas offered an ideal setting to evaluate the occurrence and variation within sediment-associated actinomycete communities. While in close geographical proximity to open reef environments, these systems provide a specialized environmental niche devoid of light and direct exposure to nutrient input. In the present study, selective isolation techniques and molecular methods were used to test the hypothesis that variable distribution of actinomycetes and secondary metabolite gene clusters occur between open reef and marine cave systems. The results indicated that differences exist within the culturable sediment-associated actinomycete communities between marine caves and open reef systems, with members of the genus Streptomyces dominating cultures from open reef sediments and a more diverse suite of actinomycetes isolated from marine cave sediment samples. Within the cave isolates, members of the proposed genus Solwaraspora were the most represented. Based on PKS- and NRPS-gene-targeted PCR amplification and sequencing, geographic variation in the occurrence of these biosynthetic pathways was also observed. These findings indicate that marine cave systems are a lucrative source in the search for novel secondary metabolite producers with biotechnological applications and that environmental and geographic factors likely affect the occurrence of these biosynthetic pathways.

  16. The Madeira Archipelago as a significant source of marine-derived actinomycete diversity with anticancer and antimicrobial potential

    Directory of Open Access Journals (Sweden)

    Alejandra Prieto-Davo

    2016-10-01

    Full Text Available Marine-derived actinomycetes have demonstrated an ability to produce novel compounds with medically relevant biological activity. Studying the diversity and biogeographical patterns of marine actinomycetes offers an opportunity to identify genera that are under environmental pressures, which may drive adaptations that yield specific biosynthetic capabilities. The present study describes research efforts to explore regions of the Atlantic Ocean, specifically around the Madeira Archipelago, where knowledge of the indigenous actinomycete diversity is scarce. A total of 400 actinomycetes were isolated, sequenced and screened for antimicrobial and anticancer activities. The three most abundant genera identified were Streptomyces, Actinomadura and Micromonospora. Phylogenetic analyses of the marine OTUs isolated indicated that the Madeira Archipelago is a new source of actinomycetes adapted to life in the ocean. Phylogenetic differences between offshore (>100m from shore and nearshore (<100m from shore populations illustrates the importance of sampling offshore in order to isolate new and diverse bacterial strains. Novel phylotypes from chemically rich marine actinomycete groups like MAR4 and the genus Salinispora were isolated. Anticancer and antimicrobial assays identified Streptomyces, Micromonospora and Salinispora as the most biologically active genera. This study illustrates the importance of bioprospecting efforts at unexplored regions of the ocean to recover bacterial strains with the potential to produce novel and interesting chemistry.

  17. The Madeira Archipelago As a Significant Source of Marine-Derived Actinomycete Diversity with Anticancer and Antimicrobial Potential

    Science.gov (United States)

    Prieto-Davó, Alejandra; Dias, Tiago; Gomes, Sofia E.; Rodrigues, Sara; Parera-Valadez, Yessica; Borralho, Pedro M.; Pereira, Florbela; Rodrigues, Cecilia M. P.; Santos-Sanches, Ilda; Gaudêncio, Susana P.

    2016-01-01

    Marine-derived actinomycetes have demonstrated an ability to produce novel compounds with medically relevant biological activity. Studying the diversity and biogeographical patterns of marine actinomycetes offers an opportunity to identify genera that are under environmental pressures, which may drive adaptations that yield specific biosynthetic capabilities. The present study describes research efforts to explore regions of the Atlantic Ocean, specifically around the Madeira Archipelago, where knowledge of the indigenous actinomycete diversity is scarce. A total of 400 actinomycetes were isolated, sequenced, and screened for antimicrobial and anticancer activities. The three most abundant genera identified were Streptomyces, Actinomadura, and Micromonospora. Phylogenetic analyses of the marine OTUs isolated indicated that the Madeira Archipelago is a new source of actinomycetes adapted to life in the ocean. Phylogenetic differences between offshore (>100 m from shore) and nearshore (marine actinomycete groups like MAR4 and the genus Salinispora were isolated. Anticancer and antimicrobial assays identified Streptomyces, Micromonospora, and Salinispora as the most biologically active genera. This study illustrates the importance of bioprospecting efforts at unexplored regions of the ocean to recover bacterial strains with the potential to produce novel and interesting chemistry. PMID:27774089

  18. Isolation and Evaluation of Marine Actinomycetes from Mangrove Forests in South of Iran against Some Human Bacterial Pathogens

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    Farshid Kafilzadeh

    2013-04-01

    Full Text Available Background & Objectives: Recent studies have shown that aquatic actinomycetes can be a source of new biological products such as antibiotics and i n dustrial products. This study was designed to examine the aquatic actinomycetes isolated from mangrove forests in South of Iran and their antibacterial activities against some human pathogens.   Methods: In this study 115 samples were randomly taken from different places of a mangrove forests in South of Iran. Isolation was based on serial dilution of the samples and plating them on starch casein agar medium. Agar well diffusion and disc diffusion assays were used to examine the antibacterial activity of the isolated purified aquatic actinomycetes.   Results: The aquatic actinomycetes were isolated from 83 samples (70%. Of them, 66 (80 percent showed antibacterial activity and 17 (20% could not inhibit the human pathogenic bacteria. The diameter of the inhibitory zones (ZOI ranged from 4 to 11 mm and the biggest zone belonged to B acillus cereus (p≤0.05.   Conclusion: The findings showed that the various and useful aquatic actinomycetes for production of new antibiotic compounds are isolated easily from the mangrove forests in South of Iran. Considering the vast spreading of mangrove forests in South of Iran and the economic and simplicity of isolation of actinomycetes for industrial usage, these source can be an important and new place for research and industry.

  19. Presence, molecular characteristics and geosmin producing ability of actinomycetes isolated from South Korean terrestrial and aquatic environments.

    Science.gov (United States)

    Lee, Gyu-Cheol; Kim, Yun S; Kim, Min-Jeong; Oh, Sung-Ae; Choi, Ilhwan; Choi, Jaewon; Park, Jong-Geun; Chong, Chom-Kyu; Kim, Yong-Yeon; Lee, Kyeunghee; Lee, Chan Hee

    2011-01-01

    The unpleasant odor of drinking water is one of the major problems in many water utilities in the world. Actinomycetes have long been associated with odorous compounds. Considering the paucity of research on Actinomycetes producing odorous compounds in South Korea, presence of Actinomycetes, their molecular characteristics and ability to produce odorous compounds were investigated in this study. Findings confirmed the presence of Actinomycetes in surface soil, sediment, and water samples from four sites: two artificial lakes [Paldang and Cheongpyeong (CP)], and two streams [Gyeongan (GA) and Yangpyeong]. Surface soil and sediment from CP area had the greatest concentration of Actinomycetes (8.2 x 10(7) and 6.8 x 10(6) colony forming units (CFUs)/gram, dry weight, respectively). When water samples are considered, samples from GA had the highest concentration (1.9 x 10(2) CFU/mL). 16S rRNA sequencing and molecular phylogenetic analysis showed that Streptomyces was the dominant genus (64.1%). In addition, the isolated Actinomycetes synthesized 5.4 ng/L geosmin as demonstrated by thermal desorption unit-gas chromatograph/mass spectrometry analysis.

  20. Three multidomain esterases from the cellulolytic rumen anaerobe Ruminococcus flavefaciens 17 that carry divergent dockerin sequences.

    Science.gov (United States)

    Aurilia, V; Martin, J C; McCrae, S I; Scott, K P; Rincon, M T; Flint, H J

    2000-06-01

    Three enzymes carrying esterase domains have been identified in the rumen cellulolytic anaerobe Ruminococcus flavefaciens 17. The newly characterized CesA gene product (768 amino acids) includes an N-terminal acetylesterase domain and an unidentified C-terminal domain, while the previously characterized XynB enzyme (781 amino acids) includes an internal acetylesterase domain in addition to its N-terminal xylanase catalytic domain. A third gene, xynE, is predicted to encode a multidomain enzyme of 792 amino acids including a family 11 xylanase domain and a C-terminal esterase domain. The esterase domains from CesA and XynB share significant sequence identity (44%) and belong to carbohydrate esterase family 3; both domains are shown here to be capable of deacetylating acetylated xylans, but no evidence was found for ferulic acid esterase activity. The esterase domain of XynE, however, shares 42% amino acid identity with a family 1 phenolic acid esterase domain identified from Clostridum thermocellum XynZ. XynB, XynE and CesA all contain dockerin-like regions in addition to their catalytic domains, suggesting that these enzymes form part of a cellulosome-like multienzyme complex. The dockerin sequences of CesA and XynE differ significantly from those previously described in R. flavefaciens polysaccharidases, including XynB, suggesting that they might represent distinct dockerin specificities.

  1. Systematic characterization of potential cellulolytic marine actinobacteria Actinoalloteichus sp. MHA15.

    Science.gov (United States)

    Rajagopal, Gobalakrishnan; Kannan, Sivakumar

    2017-03-01

    Cellulose is the most abounding biopolymer in the world and there is a great interest in using this material as a substrate for various applications and it is the most important renewable resource for bioconversion. Therefore, it is necessary to screen the cellulolytic bioorganisms. In this context, actinobacteria are one of the most efficient prokaryotes, economically and biotechnologically, for their production of about half of the discovered bioactive secondary metabolites and they can metabolize many different compounds. Therefore, the present study was carried out to isolate and screen cellulase enzyme producing marine actinobacterial strains from the sediments of the Havelock island, the Andamans. Totally, 19 morphologically distinct actinobacterial strains were isolated and subjected to cellulose degradation assay. Out of the 19, four strains were found to possess good cellulose degradation activity and the strain MHA15 alone produced higher amount of cellulase enzyme (14.379 1U/ml) than the others. Taxonomical study of the strain MHA15 revealed that it belongs to the genus Actinoalloteichus and the molecular characters showed distinct difference in its phylogenetic relationship (8.4%) with A. cyanogriseus.

  2. Systematic characterization of potential cellulolytic marine actinobacteria Actinoalloteichus sp. MHA15

    Directory of Open Access Journals (Sweden)

    Gobalakrishnan Rajagopal

    2017-03-01

    Full Text Available Cellulose is the most abounding biopolymer in the world and there is a great interest in using this material as a substrate for various applications and it is the most important renewable resource for bioconversion. Therefore, it is necessary to screen the cellulolytic bioorganisms. In this context, actinobacteria are one of the most efficient prokaryotes, economically and biotechnologically, for their production of about half of the discovered bioactive secondary metabolites and they can metabolize many different compounds. Therefore, the present study was carried out to isolate and screen cellulase enzyme producing marine actinobacterial strains from the sediments of the Havelock island, the Andamans. Totally, 19 morphologically distinct actinobacterial strains were isolated and subjected to cellulose degradation assay. Out of the 19, four strains were found to possess good cellulose degradation activity and the strain MHA15 alone produced higher amount of cellulase enzyme (14.379 1U/ml than the others. Taxonomical study of the strain MHA15 revealed that it belongs to the genus Actinoalloteichus and the molecular characters showed distinct difference in its phylogenetic relationship (8.4% with A. cyanogriseus.

  3. [Construction of Producers of Cellulolytic and Pectinolytic Enzymes Based on the Fungus Penicillium verruculosum].

    Science.gov (United States)

    Bushina, E V; Rubtsova, E A; Rozhkova, A M; Sinitsyna, O A; Koshelev, A V; Matys, V Yu; Nemashkalov, V A; Sinitsyn, A P

    2015-01-01

    Based on the fungus Penicillium verruculosum, we created strains with a complex of extracellular enzymes that contains both cellulolytic enzymes of the fungus and heterologous pectin lyase A from P. canescens and endo- 1,4-α-polygalacturonase from Aspergillus niger. The endopolygalacturonase and pectin lyase activities of enzyme preparations obtained from culture media of the producer strains reached 46-53 U/mg of protein and 1.3-2.3 U/mg of protein, respectively. The optimal temperature and pH values for recombinant pectin lyase and endopolygalacturonase corresponded to those described in the literature for these enzymes. The content of heterologous endopolygalacturonase and pectin lyase in the studied enzyme preparations was 4-5% and 23% of the total protein content, respectively. The yield of reducing sugars upon the hydrolysis of sugar beet and apple processing wastes with the most efficient preparation was 41 and 71 g/L, respectively, which corresponded to a polysaccharide conversion of 49% and 65%. Glucose was the main product of the hydrolysis of sugar beet and apple processing wastes.

  4. Paenibacillus pinihumi sp. nov., a cellulolytic bacterium isolated from the rhizosphere of Pinus densiflora.

    Science.gov (United States)

    Kim, Byung-Chun; Lee, Kang Hyun; Kim, Mi Na; Kim, Eun-Mi; Rhee, Moon-Soo; Kwon, O-Yu; Shin, Kee-Sun

    2009-10-01

    A novel cellulolytic bacterium, strain S23(T), was isolated from the rhizosphere of the pine trees in Daejeon, Republic of Korea. This isolate was Gram-positive, strictly aerobic, rod-shaped, catalase-negative, oxidase-positive, motile by means of peritrichous flagella, and tested positive for alkaline phosphatase, esterase lipase, leucine arylamidase, alpha-galactosidase, and beta-galactosidase activities. The DNA G+C content was 49.5 mol%. The main cellular fatty acids were anteiso-C(15:0) (51.9%), iso-C(16:0) (14.7%), and iso-C(15:0) (13.2%). The major isoprenoid quinone was menaquinone 7 (MK-7). Diagnostic diamino acid in the cell-wall pepti-doglycan was meso-diaminopimelic acid. Comparative 16S rRNA gene sequence analysis showed that this strain clustered with Paenibacillus species. The 16S rRNA gene sequence similarity values between S23(T) and other Paenibacillus species were between 89.9% and 95.9%, and S23(T) was most closely related to Paenibacillus tarimensis SA-7-6(T). On the basis of phylogenetic and phenotypic properties of strain S23(T), the isolate is considered as a novel species belonging to the genus Paenibacillus. Therefore, the name, Paenibacillus pinihumi sp. nov., is proposed for the rhizosphere isolate; the type strain is S23(T) (=KCTC 13695(T) =KACC 14199(T) =JCM 16419(T)).

  5. Multifunctional Cellulolytic Enzymes Outperform Processive Fungal Cellulases for Coproduction of Nanocellulose and Biofuels

    Energy Technology Data Exchange (ETDEWEB)

    Yarbrough, John M.; Zhang, Ruoran; Mittal, Ashutosh; Vander Wall, Todd; Bomble, Yannick J.; Decker, Stephen R.; Himmel, Michael E.; Ciesielski, Peter N.

    2017-03-07

    Producing fuels, chemicals, and materials from renewable resources to meet societal demands remains an important step in the transition to a sustainable, clean energy economy. The use of cellulolytic enzymes for the production of nanocellulose enables the coproduction of sugars for biofuels production in a format that is largely compatible with the process design employed by modern lignocellulosic (second generation) biorefineries. However, yields of enzymatically produced nanocellulose are typically much lower than those achieved by mineral acid production methods. In this study, we compare the capacity for coproduction of nanocellulose and fermentable sugars using two vastly different cellulase systems: the classical 'free enzyme' system of the saprophytic fungus, Trichoderma reesei (T. reesei) and the complexed, multifunctional enzymes produced by the hot springs resident, Caldicellulosiruptor bescii (C. bescii). We demonstrate by comparative digestions that the C. bescii system outperforms the fungal enzyme system in terms of total cellulose conversion, sugar production, and nanocellulose production. In addition, we show by multimodal imaging and dynamic light scattering that the nanocellulose produced by the C. bescii cellulase system is substantially more uniform than that produced by the T. reesei system. These disparities in the yields and characteristics of the nanocellulose produced by these disparate systems can be attributed to the dramatic differences in the mechanisms of action of the dominant enzymes in each system.

  6. Cellulosic Ethanol Production by Recombinant Cellulolytic Bacteria Harbouring pdc and adh II Genes of Zymomonas mobilis.

    Science.gov (United States)

    Piriya, P Sobana; Vasan, P Thirumalai; Padma, V S; Vidhyadevi, U; Archana, K; Vennison, S John

    2012-01-01

    The ethanol fermenting genes such as pyruvate decarboxylase (pdc) and alcohol dehydrogenase II (adh II) were cloned from Zymomonas mobilis and transformed into three different cellulolytic bacteria, namely Enterobacter cloacae JV, Proteus mirabilis JV and Erwinia chrysanthemi and their cellulosic ethanol production capability was studied. Recombinant E. cloacae JV was found to produce 4.5% and 3.5% (v/v) ethanol, respectively, when CMC and 4% NaOH pretreated bagasse were used as substrates, whereas recombinant P. mirabilis and E. chrysanthemi with the same substrates could only produce 4%, 3.5%, 1%, and 1.5 % of ethanol, respectively. The recombinant E. cloacae strain produced twofold higher percentage of ethanol than the wild type. The recombinant E. cloacae strain could be improved further by increasing its ethanol tolerance capability through media optimization and also by combining multigene cellulase expression for enhancing ethanol production from various types of lignocellulosic biomass so that it can be used for industrial level ethanol production.

  7. Bacteroides luti sp. nov., an anaerobic, cellulolytic and xylanolytic bacterium isolated from methanogenic sludge.

    Science.gov (United States)

    Hatamoto, Masashi; Kaneshige, Masami; Nakamura, Akinobu; Yamaguchi, Takashi

    2014-05-01

    A mesophilic, anaerobic, cellulolytic and xylanolytic strain, UasXn-3T, was isolated from anaerobic granular sludge in a mesophilic upflow anaerobic sludge blanket reactor, which was used to treat municipal sewage. The cells were Gram-stain-negative, non-motile, and non-spore-forming rods. The optimal temperature for growth was 37-40 °C and the optimal pH for growth was pH 6.5-7.0. Strain UasXn-3T could grow on several polysaccharides and sugars, including cellulose, cellobiose, xylan, xylose, glucose, fructose, arabinose, mannose, raffinose, trehalose and starch. The DNA G+C content was 44.4 mol%. On the basis of comparative 16S rRNA gene sequence analysis, strain UasXn-3T was identified as a member of the genus Bacteroides and most closely related to Bacteroides oleiciplenus, B. intestinalis, B. cellulosilyticus and B. graminisolvens (sequence similarities of 91.3-91.6%). Since the genetic and phenotypic properties suggest that strain UasXn-3T represents a novel species, we propose the name Bacteroides luti sp. nov. The type strain is UasXn-3T (=JCM 19020T=DSM 26991T).

  8. Bacteroides cellulosilyticus sp. nov., a cellulolytic bacterium from the human gut microbial community.

    Science.gov (United States)

    Robert, Céline; Chassard, Christophe; Lawson, Paul A; Bernalier-Donadille, Annick

    2007-07-01

    A strictly anaerobic cellulolytic bacterium, strain CRE21(T), was isolated from a human faecal sample. Cells were Gram-negative non-motile rods that were about 1.7 microm in length and 0.9 microm in width. Strain CRE21(T) degraded different types of cellulose and was able to grow on a variety of carbohydrates. Cellulose and sugars were mainly converted to acetate, propionate and succinate. The G+C content of the DNA was 41.1 mol%. 16S rRNA gene sequence analysis revealed that the isolate belonged to the genus Bacteroides with highest sequence similarity to the type strain of Bacteroides intestinalis (98 %). DNA-DNA hybridization results revealed that strain CRE21(T) was distinct from B. intestinalis (40 % DNA-DNA relatedness). Strain CRE21(T) also showed several characteristics distinct from B. intestinalis. In particular, it exhibited different capacity to degrade polysaccharides such as cellulose. On the basis of phylogenetic analysis and the morphological, physiological and biochemical data presented in this study, strain CRE21(T) can be readily differentiated from recognized species of the genus Bacteroides. The name Bacteroides cellulosilyticus sp. nov. is proposed to accommodate this organism. The type strain is CRE21(T) (=DSM 14838(T)=CCUG 44979(T)).

  9. Cellulosic Ethanol Production by Recombinant Cellulolytic Bacteria Harbouring pdc and adh II Genes of Zymomonas mobilis

    Directory of Open Access Journals (Sweden)

    P. Sobana Piriya

    2012-01-01

    Full Text Available The ethanol fermenting genes such as pyruvate decarboxylase (pdc and alcohol dehydrogenase II (adh II were cloned from Zymomonas mobilis and transformed into three different cellulolytic bacteria, namely Enterobacter cloacae JV, Proteus mirabilis JV and Erwinia chrysanthemi and their cellulosic ethanol production capability was studied. Recombinant E. cloacae JV was found to produce 4.5% and 3.5% (v/v ethanol, respectively, when CMC and 4% NaOH pretreated bagasse were used as substrates, whereas recombinant P. mirabilis and E. chrysanthemi with the same substrates could only produce 4%, 3.5%, 1%, and 1.5 % of ethanol, respectively. The recombinant E. cloacae strain produced twofold higher percentage of ethanol than the wild type. The recombinant E. cloacae strain could be improved further by increasing its ethanol tolerance capability through media optimization and also by combining multigene cellulase expression for enhancing ethanol production from various types of lignocellulosic biomass so that it can be used for industrial level ethanol production.

  10. Quantitative isolation of biocontrol agents Trichoderma spp., Gliocladium spp. and actinomycetes from soil with culture media.

    Science.gov (United States)

    Vargas Gil, S; Pastor, S; March, G J

    2009-01-01

    Soil biodiversity plays a key role in the sustainability of agriculture systems and indicates the level of health of soil, especially when considering the richness of microorganisms that are involved in biological control of soilborne diseases. Cultural practices may produce changes in soil microflora, which can be quantified through the isolation of target microorganisms. Rhizosphere soil samples were taken from an assay with different crop rotations and tillage systems, and populations of Trichoderma spp., Gliocladium spp. and actinomycetes were quantified in order to select the general and selective culture media that better reflect the changes of these microbial populations in soil. The most efficient medium for the isolation of Trichoderma spp. and Gliocladium spp. was potato dextrose agar modified by the addition of chloramphenicol, streptomycin and rose bengal, and for actinomycetes was Küster medium, with cycloheximide and sodium propionate.

  11. ANTIBIOTIC COMPOUND FROM MARINE ACTINOMYCETES (Streptomyces sp A11: ISOLATION AND STRUCTURE ELUCIDATON

    Directory of Open Access Journals (Sweden)

    Rofiq Sunaryanto

    2010-07-01

    Full Text Available Purification and structure elucidation of antibiotic produced by marine actinomycetes (Streptomyces sp A11 was conducted. Production of antibiotic was carried out by liquid fermentation using yeast and peptone medium for 5 days fermentation. Purification of antibiotic was carried out by silica gel 60 (Merck, 0.063-0.200 mm column chromatography and preparative HPLC. Structure elucidation was carried out using ESI-MS, 1H NMR, 13C NMR, DEPT 13C NMR, and FTIR. This antibiotic was identified as cyclo (tyrosyl-prolyl / (C14H16N2O3. This antibiotic had biological activity to Escherichia coli ATCC 25922, Staphylococcus aureus ATCC25923, Bacillus subtilis ATCC 66923, Pseudomonas aeruginosa ATCC27853, and produced by extracellular secretion.   Keywords: antibiotic, actinomycetes, purification, structure elucidation

  12. Isolation and partial characterization of actinomycetes with antimicrobial activity against multidrug resistant bacteria

    Institute of Scientific and Technical Information of China (English)

    Smriti Singh; Pramod Kumar; N Gopalan; Bhuvnesh Shrivastava; RC Kuhad; Hotam Singh Chaudhary

    2012-01-01

    Objective: To isolate strains of Actinomycetes from different locations of Gwalior to evaluate its antimicrobial activity against multidrug resistant pathogenic strains. Method: Soil samples collected from different niche habitats of Gwalior were serially diluted and plated on selective media. Potential colonies were further purified and stored in agar slants and glycerol stocks. Isolates were biochemically characterized and purified isolates were test against pathogenic microorganisms for screening. Isolates with antagonistic properties were inoculated in production media and secondary metabolites or antimicrobial products were extracted. Result: The seven actinomycetes strains showing maximum antibacterial activity were isolated further characterized based on their colony characteristics and biochemical analyses. The isolates were screened for their secondary metabolites activity on three human pathogenic bacteria are Escherichia coli (E. coli), Methicillin-Resistant Staphylococcus aureus (S. aureus) and Vancomycin-Resistant Enterococci (VRE). Discussion: The strain MITS 1005 was found to be more active against the test bacteria.

  13. Volatile terpenes from actinomycetes: a biosynthetic study correlating chemical analyses to genome data.

    Science.gov (United States)

    Rabe, Patrick; Citron, Christian A; Dickschat, Jeroen S

    2013-11-25

    The volatile terpenes of 24 actinomycetes whose genomes have been sequenced (or are currently being sequenced) were collected by use of a closed-loop stripping apparatus and identified by GC/MS. The analytical data were compared against a phylogenetic analysis of all 192 currently available sequences of bacterial terpene cyclases (excluding geosmin and 2-methylisoborneol synthases). In addition to the several groups of terpenes with known biosynthetic origin, selinadienes were identified as a large group of biosynthetically related sesquiterpenes that are produced by several streptomycetes. The detection of a large number of previously unrecognised side products of known terpene cyclases proved to be particularly important for an in depth understanding of biosynthetic pathways to known terpenes in actinomycetes. Interpretation of the chemical analytical data in the context of the phylogenetic tree of bacterial terpene cyclases pointed to the function of three new enzymes: (E)-β-caryophyllene synthase, selina-3,7(11)-diene synthase and aristolochene synthase.

  14. Inhibition of norsolorinic acid accumulation to Aspergillus parasiticus by marine actinomycetes

    Science.gov (United States)

    Yan, Peisheng; Shi, Cuijuan; Shen, Jihong; Wang, Kai; Gao, Xiujun; Li, Ping

    2014-11-01

    Thirty-six strains of marine actinomycetes were isolated from a sample of marine sediment collected from the Yellow Sea and evaluated in terms of their inhibitory activity on the growth of Aspergillus parasiticus and the production of norsolorinic acid using dual culture plate assay and agar diffusion methods. Among them, three strains showed strong antifungal activity and were subsequently identified as Streptomyces sp. by 16S rRNA gene sequencing analysis. The supernatant from the fermentation of the MA01 strain was extracted sequentially with chloroform and ethyl acetate, and the activities of the extracts were determined by tip culture assay. The assay results show that both extracts inhibited mycelium growth and toxin production, and the inhibitory activities of the extracts increased as their concentrations increased. The results of this study suggest that marine actinomycetes are biologically important for the control of mycotoxins, and that these bacteria could be used as novel biopesticides against mycotoxins.

  15. Underground Cordon by Microorganisms-Part-III Role of Soil Inhabiting Actinomycetes

    Directory of Open Access Journals (Sweden)

    H. M. Dayal

    1989-04-01

    Full Text Available Certain strains of soil inhabiting actinomycetes were found to substantially corrode aluminium alloy (54-S which has bscn found tobe more resistant to bacterial or fungal corrosion in our earlier studies.These strains did not produce any corrosion on the mild steel and galvanised iron panels which were heavily corroded by bacteria and fungi. The corrosive isolates have been partialiy characterised after their isolation and purification. The extent of corrosion caused by eachstrain has been determined.

  16. Halophilic and halotolerant actinomycetes from a marine saltern of Goa, India producing anti-bacterial metabolites.

    Science.gov (United States)

    Ballav, Shuvankar; Kerkar, Savita; Thomas, Sabu; Augustine, Nimmy

    2015-03-01

    Marine salterns are estuarine ecosystems in Goa, receiving inputs from riverine and marine waters. The Salinity fluctuates between 0 and 300 psu which makes it a conducive niche for salt tolerant and salt loving Actinomycetales. Halotolerant and halophilic Actinomycetales producing anti-bacterial metabolites were studied from crystallizer pond sediments of Ribandar saltern, Goa. Three media viz. Starch casein, R2A and Inorganic salt starch agar at four different salinities (35, 50, 75 and 100 psu) were used for isolation. R2A agar at 35 psu was the most preferred by hypersaline actinomycetes. The dominant group was halotolerant Streptomyces spp. others being rare actinomycetes viz. Nocardiopsis, Micromonospora and Kocuria spp. More than 50% of the isolates showed anti-bacterial activity against one or more of the fifteen human pathogens tested. Eight strains from 4 genera showed consistent anti-bacterial activity and studied in detail. Most halotolerant isolates grew from 0 to 75 psu, with optimum antibiotic production at 35 psu whereas halophiles grew at 20 to 100 psu with optimum antibiotic production at 35 psu. Four Streptomyces strains showed multiple inhibition against test organisms while four rare actinomycetes were specific in their inhibitory activity. This is the first report of a halophilic Kocuria sp., Nocardiopsis sp., and halotolerant Micromonospora sp. producing anti-bacterial compound(s) against Staphylococcus aureus, Staphylococcus citreus, and Vibrio cholerae, respectively. Sequential extraction with varying polarity of organic solvents showed that the extracts inhibited different test pathogens. These results suggest that halophilic and halotolerant actinomycetes from marine salterns are a potential source of anti-bacterial compounds. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  17. Marine Actinomycetes as potential source for histone deacetylase inhibitors and epigenetic modulation.

    Science.gov (United States)

    Varghese, T A; Jayasri, M A; Suthindhiran, K

    2015-07-01

    In the light of important detrimental role of aberrant histone deacetylases (HDAC) production during various clinical complications, development of therapeutically effective and specific inhibitors of HDAC is critically important. This study deals with the screening for HDAC inhibitors from marine Actinomycetes. The isolation of Actinomycetes from 22 sediment samples along the Southern Coast of India yielded 186 strains including Streptomyces, Nocardipsis, evaluated for HDAC inhibition using HeLa cells. Among the 186 isolates, 10 strains have shown moderate to strong inhibition. The maximum inhibition (61%) was seen with strain VITKSM06 and least inhibition (31%) was seen with strain VITSJT03. The MTT cell proliferation assay using HeLa cell line showed significant cytotoxicity with an IC50 of 5·9 μg ml(-1) by VITKSM06-derived metabolite and 26·2 μg ml(-1) by VITSJT03. The compound treated HeLa cells displayed an altered morphology and condensed chromatin which may be due to HDAC inhibition. Based on the phylogenetic analysis, the potential strains were identified as Nocardiopsis sp VITKSM06, Streptomyces sp VITAKS1 and Streptomyces sp VITRSM02. This study reveals the importance of screening marine Actinomycetes for the discovery of potential novel HDAC inhibitors of therapeutic importance. Histone deacetylases (HDAC) are epigenetic enzymes that regulate the deacetylation in lysine group on a histone, and thus regulate the gene expression. The HDAC inhibitors are reported to promote apoptosis on tumour cells, thus become clinically important drug target. Several studies have addressed the identification of putative HDAC inhibitors as therapeutic agents for cancer and until now those cleared phase III human trials are very limited. This study attempts to investigate the chemical diversity found in marine Actinomycetes towards negative HDAC modulation, which could be used individually or in combination as anti-cancerous and other therapeutic measure. © 2015 The

  18. Three New 2-pyranone Derivatives from Mangrove Endophytic Actinomycete Strain Nocardiopsis sp. A00203

    Directory of Open Access Journals (Sweden)

    Yuemao Shen

    2010-10-01

    Full Text Available Three new 2-pyranone derivatives, namely Norcardiatones A (1, B (2 and C (3, were isolated from the agar cultures of the strain Nocardiopsis sp. A00203, a mangrove endophytic actinomycete. Their structures were elucidated by spectroscopic and mass-spectrometric analyses, including 1D-, 2D-NMR and HR Q-TOF-MS. Compound 1 showed week cytotoxicity against HeLa cells in MTT assay.

  19. A mixed community of actinomycetes produce multiple antibiotics for the fungus farming ant Acromyrmex octospinosus

    OpenAIRE

    Barke Jörg; Seipke Ryan F; Grüschow Sabine; Heavens Darren; Drou Nizar; Bibb Mervyn J; Goss Rebecca JM; Yu Douglas W; Hutchings Matthew I

    2010-01-01

    This work was supported by a UEA-funded PhD studentship (JB) and an MRC Milstein award, G0801721 (MIH, RJMG and DY). MIH is a Research Councils UK Fellow. DY also received support from the Yunnan provincial government (20080A001) and the Chinese Academy of Sciences (0902281081). Background: Attine ants live in an intensely studied tripartite mutualism with the fungus Leucoagaricus gongylophorus, which provides food to the ants, and with antibiotic-producing actinomycete bacteria. One hypot...

  20. Conservation of thiol-oxidative stress responses regulated by SigR orthologues in actinomycetes

    Science.gov (United States)

    Kim, Min-Sik; Dufour, Yann S.; Yoo, Ji Sun; Cho, Yoo-Bok; Park, Joo-Hong; Nam, Gi-Baeg; Kim, Hae Min; Lee, Kang-Lok; Donohue, Timothy J.; Roe, Jung-Hye

    2015-01-01

    Summary Numerous thiol-reactive compounds cause oxidative stress where cells counteract by activation of survival strategies regulated by thiol-based sensors. In Streptomyces coelicolor, a model actinomycete, a sigma/antisigma pair SigR/RsrA controls the response to thiol-oxidative stress. To unravel its full physiological functions, chromatin immuno-precipitation combined with sequence and transcript analyses were employed to identify 108 SigR target genes in S. coelicolor and to predict orthologous regulons across actinomycetes. In addition to reported genes for thiol homeostasis, protein degradation and ribosome modulation, 64 additional operons were identified suggesting new functions of this global regulator. We demonstrate that SigR maintains the level and activity of the housekeeping sigma factor HrdB during thiol-oxidative stress, a novel strategy for stress responses. We also found that SigR defends cells against UV and thiol-reactive damages, in which repair UvrA takes a part. Using a refined SigR-binding sequence model, SigR orthologues and their targets were predicted in 42 actinomycetes. This revealed a conserved core set of SigR targets to function for thiol homeostasis, protein quality control, possible modulation of transcription and translation, flavin-mediated redox reactions, and Fe-S delivery. The composition of the SigR regulon reveals a robust conserved physiological mechanism to deal with thiol-oxidative stress from bacteria to human. PMID:22651816

  1. Actinomycete complexes in soils of industrial and residential zones in the city of Kirov

    Science.gov (United States)

    Shirokikh, I. G.; Solov'eva, E. S.; Ashikhmina, T. Ya.

    2014-02-01

    The number, diversity, and structure of the actinomycetal complexes in the soils of the industrial and residential zones of the city of Kirov are considered. The total content of mobile cadmium, zinc, lead, iron, and nickel in the soils of the industrial biotopes was 1.8 and 6.0 times higher than their concentration in the soils of the residential and background zones, respectively. In the heavy metal (HM)-polluted soils, the share of actinomycetes in the total number of prokaryotes and the relative abundance of the micromono-spores in the actinomycetal complex were much higher and the species diversity of the streptomycetes was lower than these characteristics in the soils of the residential zone. The differences in the composition of the mycelial prokaryote complexes appear to be related to the selective resistance of some of their representatives to heavy metals. The possibility to select the strains resistant to HMs and suitable for use in the bioremediation of polluted soils is considered.

  2. Diversity and biosynthetic potential of culturable actinomycetes associated with marine sponges in the China Seas.

    Science.gov (United States)

    Xi, Lijun; Ruan, Jisheng; Huang, Ying

    2012-01-01

    The diversity and secondary metabolite potential of culturable actinomycetes associated with eight different marine sponges collected from the South China Sea and the Yellow sea were investigated. A total of 327 strains were isolated and 108 representative isolates were selected for phylogenetic analysis. Ten families and 13 genera of Actinomycetales were detected, among which five genera represent first records isolated from marine sponges. Oligotrophic medium M5 (water agar) proved to be efficient for selective isolation, and "Micromonospora-Streptomyces" was proposed as the major distribution group of sponge-associated actinomycetes from the China Seas. Ten isolates are likely to represent novel species. Sponge Hymeniacidon perleve was found to contain the highest genus diversity (seven genera) of actinomycetes. Housekeeping gene phylogenetic analyses of the isolates indicated one ubiquitous Micromonospora species, one unique Streptomyces species and one unique Verrucosispora phylogroup. Of the isolates, 27.5% displayed antimicrobial activity, and 91% contained polyketide synthase and/or nonribosomal peptide synthetase genes, indicating that these isolates had a high potential to produce secondary metabolites. The isolates from sponge Axinella sp. contained the highest presence of both antimicrobial activity and NRPS genes, while those from isolation medium DNBA showed the highest presence of antimicrobial activity and PKS I genes.

  3. Sequencing rare marine actinomycete genomes reveals high density of unique natural product biosynthetic gene clusters.

    Science.gov (United States)

    Schorn, Michelle A; Alanjary, Mohammad M; Aguinaldo, Kristen; Korobeynikov, Anton; Podell, Sheila; Patin, Nastassia; Lincecum, Tommie; Jensen, Paul R; Ziemert, Nadine; Moore, Bradley S

    2016-12-01

    Traditional natural product discovery methods have nearly exhausted the accessible diversity of microbial chemicals, making new sources and techniques paramount in the search for new molecules. Marine actinomycete bacteria have recently come into the spotlight as fruitful producers of structurally diverse secondary metabolites, and remain relatively untapped. In this study, we sequenced 21 marine-derived actinomycete strains, rarely studied for their secondary metabolite potential and under-represented in current genomic databases. We found that genome size and phylogeny were good predictors of biosynthetic gene cluster diversity, with larger genomes rivalling the well-known marine producers in the Streptomyces and Salinispora genera. Genomes in the Micrococcineae suborder, however, had consistently the lowest number of biosynthetic gene clusters. By networking individual gene clusters into gene cluster families, we were able to computationally estimate the degree of novelty each genus contributed to the current sequence databases. Based on the similarity measures between all actinobacteria in the Joint Genome Institute's Atlas of Biosynthetic gene Clusters database, rare marine genera show a high degree of novelty and diversity, with Corynebacterium, Gordonia, Nocardiopsis, Saccharomonospora and Pseudonocardia genera representing the highest gene cluster diversity. This research validates that rare marine actinomycetes are important candidates for exploration, as they are relatively unstudied, and their relatives are historically rich in secondary metabolites.

  4. Isolation and in vitro selection of actinomycetes strains as potential probiotics for aquaculture

    Directory of Open Access Journals (Sweden)

    Milagro García Bernal

    2015-02-01

    Full Text Available Aim: This study was designed to describe a series of in vitro tests that may aid the discovery of probiotic strains from actinomycetes. Materials and Methods: Actinomycetes were isolated from marine sediments using four different isolation media, followed by antimicrobial activity and toxicity assessment by the agar diffusion method and the hemolysis of human blood cells, respectively. Extracellular enzymatic production was monitored by the hydrolysis of proteins, lipids and carbohydrates. Tolerance to different pH values and salt concentrations was also determined, followed by hydrophobicity analysis and genetic identification of the most promising strains. Results: Five out of 31 isolated strains showed antimicrobial activity against three Vibrio species. Three non-hemolytic strains (N7, RL8 and V4 among these active isolates yielded positive results in hydrophobicity tests and exhibited good growth at salt concentrations ranging from 0% to 10%, except strain RL8, which required a salt concentration >0.6%. Although these strains did not grow at pH<3, they showed different enzymatic activities. Phylogenetic analysis revealed that strains N7 and V4 have more than 99% identity with several Streptomyces species, whereas the closest matches to strain RL8 are Streptomyces panacagri and Streptomyces flocculus, with 98% and 98.2% similarity, respectively. Conclusion: Three actinomycetes strains showing probiotic-like properties were discovered using several in vitro tests that can be easily implemented in different institutions around the world.

  5. Diversity and Biosynthetic Potential of Culturable Actinomycetes Associated with Marine Sponges in the China Seas

    Directory of Open Access Journals (Sweden)

    Ying Huang

    2012-05-01

    Full Text Available The diversity and secondary metabolite potential of culturable actinomycetes associated with eight different marine sponges collected from the South China Sea and the Yellow sea were investigated. A total of 327 strains were isolated and 108 representative isolates were selected for phylogenetic analysis. Ten families and 13 genera of Actinomycetales were detected, among which five genera represent first records isolated from marine sponges. Oligotrophic medium M5 (water agar proved to be efficient for selective isolation, and “MicromonosporaStreptomyces” was proposed as the major distribution group of sponge-associated actinomycetes from the China Seas. Ten isolates are likely to represent novel species. Sponge Hymeniacidon perleve was found to contain the highest genus diversity (seven genera of actinomycetes. Housekeeping gene phylogenetic analyses of the isolates indicated one ubiquitous Micromonospora species, one unique Streptomyces species and one unique Verrucosispora phylogroup. Of the isolates, 27.5% displayed antimicrobial activity, and 91% contained polyketide synthase and/or nonribosomal peptide synthetase genes, indicating that these isolates had a high potential to produce secondary metabolites. The isolates from sponge Axinella sp. contained the highest presence of both antimicrobial activity and NRPS genes, while those from isolation medium DNBA showed the highest presence of antimicrobial activity and PKS I genes.

  6. Taxonomy and Polyphasic Characterization of Alkaline Amylase Producing Marine Actinomycete Streptomyces rochei BTSS 1001

    Directory of Open Access Journals (Sweden)

    Aparna Acharyabhatta

    2013-01-01

    Full Text Available Actinomycetes isolated from marine sediments along the southeast coast of Bay of Bengal were investigated for amylolytic activity. Marine actinomycete BTSS 1001 producing an alkaline amylase was identified from marine sediment of Diviseema coast, Bay of Bengal. The isolate produced alkaline amylase with maximum amylolytic activity at pH 9.5 at 50°C. The organism produced white to pale grey substrate mycelium and grayish aerial mycelium with pinkish brown pigmentation. A comprehensive study of morphological, physiological parameters, cultural characteristics, and biochemical studies was performed. The presence of iso-C15 : 0, anteiso-C15 : 0, iso-C16 : 0, and anteiso-C17 : 0 as the major cellular fatty acids, LL-diaminopimelic acid as the characteristic cell wall component, and menaquinones MK-9H(6 and MK-9H(8 as the major isoprenoid quinones is attributed to the strain BTSS 1001 belonging to the genus Streptomyces. Comparison of 16S rRNA gene sequences showed that strain BTSS 1001 exhibited the highest similarities to the type strains of Streptomyces rochei (99%, Streptomyces plicatus (99%, and Streptomyces enissocaesilis (99%. Using the polyphasic taxonomical approach and phenotypic characteristic studies, the isolate BTSS 1001 was characterized as marine actinomycete Streptomyces rochei.

  7. Actinomycetes: A Repertory of Green Catalysts with a Potential Revenue Resource

    Science.gov (United States)

    Prakash, Divya; Nawani, Neelu; Prakash, Mansi; Bodas, Manish; Mandal, Abul; Khetmalas, Madhukar; Kapadnis, Balasaheb

    2013-01-01

    Biocatalysis, one of the oldest technologies, is becoming a favorable alternative to chemical processes and a vital part of green technology. It is an important revenue generating industry due to a global market projected at $7 billion in 2013 with a growth of 6.7% for enzymes alone. Some microbes are important sources of enzymes and are preferred over sources of plant and animal origin. As a result, more than 50% of the industrial enzymes are obtained from bacteria. The constant search for novel enzymes with robust characteristics has led to improvisations in the industrial processes, which is the key for profit growth. Actinomycetes constitute a significant component of the microbial population in most soils and can produce extracellular enzymes which can decompose various materials. Their enzymes are more attractive than enzymes from other sources because of their high stability and unusual substrate specificity. Actinomycetes found in extreme habitats produce novel enzymes with huge commercial potential. This review attempts to highlight the global importance of enzymes and extends to signify actinomycetes as promising harbingers of green technology. PMID:23691495

  8. Isolation and in vitro selection of actinomycetes strains as potential probiotics for aquaculture

    Science.gov (United States)

    Bernal, Milagro García; Campa-Córdova, Ángel Isidro; Saucedo, Pedro Enrique; González, Marlen Casanova; Marrero, Ricardo Medina; Mazón-Suástegui, José Manuel

    2015-01-01

    Aim: This study was designed to describe a series of in vitro tests that may aid the discovery of probiotic strains from actinomycetes. Materials and Methods: Actinomycetes were isolated from marine sediments using four different isolation media, followed by antimicrobial activity and toxicity assessment by the agar diffusion method and the hemolysis of human blood cells, respectively. Extracellular enzymatic production was monitored by the hydrolysis of proteins, lipids and carbohydrates. Tolerance to different pH values and salt concentrations was also determined, followed by hydrophobicity analysis and genetic identification of the most promising strains. Results: Five out of 31 isolated strains showed antimicrobial activity against three Vibrio species. Three non-hemolytic strains (N7, RL8 and V4) among these active isolates yielded positive results in hydrophobicity tests and exhibited good growth at salt concentrations ranging from 0% to 10%, except strain RL8, which required a salt concentration >0.6%. Although these strains did not grow at pH<3, they showed different enzymatic activities. Phylogenetic analysis revealed that strains N7 and V4 have more than 99% identity with several Streptomyces species, whereas the closest matches to strain RL8 are Streptomyces panacagri and Streptomyces flocculus, with 98% and 98.2% similarity, respectively. Conclusion: Three actinomycetes strains showing probiotic-like properties were discovered using several in vitro tests that can be easily implemented in different institutions around the world. PMID:27047067

  9. Eco-taxonomic insights into actinomycete symbionts of termites for discovery of novel bioactive compounds.

    Science.gov (United States)

    Kurtböke, D Ipek; French, John R J; Hayes, R Andrew; Quinn, Ronald J

    2015-01-01

    Termites play a major role in foraging and degradation of plant biomass as well as cultivating bioactive microorganisms for their defense. Current advances in "omics" sciences are revealing insights into function-related presence of these symbionts, and their related biosynthetic activities and genes identified in gut symbiotic bacteria might offer a significant potential for biotechnology and biodiscovery. Actinomycetes have been the major producers of bioactive compounds with an extraordinary range of biological activities. These metabolites have been in use as anticancer agents, immune suppressants, and most notably, as antibiotics. Insect-associated actinomycetes have also been reported to produce a range of antibiotics such as dentigerumycin and mycangimycin. Advances in genomics targeting a single species of the unculturable microbial members are currently aiding an improved understanding of the symbiotic interrelationships among the gut microorganisms as well as revealing the taxonomical identity and functions of the complex multilayered symbiotic actinofloral layers. If combined with target-directed approaches, these molecular advances can provide guidance towards the design of highly selective culturing methods to generate further information related to the physiology and growth requirements of these bioactive actinomycetes associated with the termite guts. This chapter provides an overview on the termite gut symbiotic actinoflora in the light of current advances in the "omics" science, with examples of their detection and selective isolation from the guts of the Sunshine Coast regional termite Coptotermes lacteus in Queensland, Australia.

  10. Occupational allergic respiratory diseases in garbage workers: relevance of molds and actinomycetes.

    Science.gov (United States)

    Hagemeyer, O; Bünger, J; van Kampen, V; Raulf-Heimsoth, M; Drath, C; Merget, R; Brüning, Th; Broding, H C

    2013-01-01

    Exposures to molds and bacteria (especially actinomycetes) at workplaces are common in garbage workers, but allergic respiratory diseases due to these microorganisms have been described rarely. The aim of our study was a detailed analysis of mold or bacteria-associated occupational respiratory diseases in garbage workers. From 2002 to 2011 four cases of occupational respiratory diseases related to garbage handling were identified in our institute (IPA). Hypersensitivity pneumonitis (HP) was diagnosed in three subjects (cases 1-3, one smoker, two non-smokers), occupational asthma (OA) was diagnosed in one subject (case 4, smoker), but could not be excluded completely in case 2. Cases 1 and 2 worked in composting sites, while cases 3 and 4 worked in packaging recycling plants. Exposure periods were 2-4 years. Molds and actinomycetes were identified as allergens in all cases. Specific IgE antibodies to Aspergillus fumigatus were detected exclusively in case 4. Diagnoses of HP were essentially based on symptoms and the detection of specific IgG serum antibodies to molds and actinomycetes. OA was confirmed by bronchial provocation test with Aspergillus fumigatus in case 4. In conclusion, occupational HP and OA due to molds occur rarely in garbage workers. Technical prevention measures are insufficient and the diagnosis of HP is often inconclusive. Therefore, it is recommended to implement the full repertoire of diagnostic tools including bronchoalveolar lavage and high resolution computed tomography in the baseline examination.

  11. Actinomycetes: a repertory of green catalysts with a potential revenue resource.

    Science.gov (United States)

    Prakash, Divya; Nawani, Neelu; Prakash, Mansi; Bodas, Manish; Mandal, Abul; Khetmalas, Madhukar; Kapadnis, Balasaheb

    2013-01-01

    Biocatalysis, one of the oldest technologies, is becoming a favorable alternative to chemical processes and a vital part of green technology. It is an important revenue generating industry due to a global market projected at $7 billion in 2013 with a growth of 6.7% for enzymes alone. Some microbes are important sources of enzymes and are preferred over sources of plant and animal origin. As a result, more than 50% of the industrial enzymes are obtained from bacteria. The constant search for novel enzymes with robust characteristics has led to improvisations in the industrial processes, which is the key for profit growth. Actinomycetes constitute a significant component of the microbial population in most soils and can produce extracellular enzymes which can decompose various materials. Their enzymes are more attractive than enzymes from other sources because of their high stability and unusual substrate specificity. Actinomycetes found in extreme habitats produce novel enzymes with huge commercial potential. This review attempts to highlight the global importance of enzymes and extends to signify actinomycetes as promising harbingers of green technology.

  12. Mesophilic Actinomycetes in the natural and reconstructed sand dune vegetation zones of Fraser Island, Australia.

    Science.gov (United States)

    Kurtböke, D I; Neller, R J; Bellgard, S E

    2007-08-01

    The natural coastal habitat of Fraser Island located in the State of Queensland, Australia, has been disturbed in the past for mining of the mineral sand ilmenite. Currently, there is no information available on whether these past mining disturbances have affected the distribution, diversity, and survival of beneficial soil microorganisms in the sand dunes of the island. This in turn could deleteriously affect the success of the natural regeneration, plant growth, and establishment on the sand dunes. To support ongoing restoration efforts at sites like these mesophilic actinomycetes were isolated using conventional techniques, with particular emphasis on the taxa previously reported to produce plant-growth-promoting substances and providing support to mycorrhizal fungi, were studied at disturbed sites and compared with natural sites. In the natural sites, foredunes contained higher densities of micromonosporae replaced by increasing numbers of streptomycete species in the successional dune and finally leading to complex actinomycete communities in the mature hind dunes. Whereas in the disturbed zones affected by previous mining activities, which are currently being rehabilitated, no culturable actinomycete communities were detected. These findings suggest that the paucity of beneficial microflora in the rehabilitated sand dunes may be limiting the successful colonization by pioneer plant species. Failure to establish a cover of plant species would result in the mature hind dune plants being exposed to harsh salt and climatic conditions. This could exacerbate the incidence of wind erosion, resulting in the destabilization of well-defined and vegetated successional dunal zones.

  13. The prevalence of actinomycetes-like organisms found in cervicovaginal smears of 300 IUD wearers.

    Science.gov (United States)

    Jones, M C; Buschmann, B O; Dowling, E A; Pollock, H M

    1979-01-01

    The association of Actinomyces with IUD wearers has been widely documented and the possibility of the recognition of actinomycetes-like organisms in routine Papanicolaou-stained cervicovaginal smears has been reported. We conducted a retrospective study of IUD wearers to determine the prevalence and significance of actinomycetes-like organisms found in such smears. Three hundred smears from current IUD wearers were rescreened for actinomycetes-like organisms. Of this group, 200 patients were from a public health family planning clinic, and 100 were private patients. The incidence for the public health group was 25.5% and for the private patient group, 8%. A case history of actinomycosis is included. Findings such as other infectious agents, abnormal cytology and symptoms are also discussed. Although the presence of Actinomyces probably represents an opportunistic infection, the threat of pelvic actinomycosis with serious complications poses a management problem to the clinician when Actinomyces is reported in a routine Papanicolaou smear. Our findings lead us to question the practicality of the earlier recommendations of IUD removal and antibiotic therapy.

  14. Bioperspective of actinomycetes isolates from coastal soils: A new source of antimicrobial producers

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    Rattanaporn Srivibool

    2006-05-01

    Full Text Available Forty five soil samples were collected from four coastal islands on the east coast of Thailand: Chang, Hwai, Lao-yanai in Trat Province and Pai Islands in Chonburi Province. On 3 isolating media, Actinomycetes Isolation Agar, Starch Casein Agar and Glucose Asparagine Agar, 495 isolates of actinomycetes were found. Preliminary test to search for antimicrobial activity was done with Bacillus subtilis TISTR 008, Staphylococcus aureus TISTR 885, Staphylococus aureus TISTR 517 (ATCC 25923, Micrococcus luteus TISTR 884 and Pseudomonas aeruginosa TISTR 781 and Escherichia coli TISTR 887 (ATCC 25922. Fifty-eight actinomycetes were found to be antimicrobial-producing strains. From the morphological determination, cell wall diaminopimelic acid and sugars in whole-cell hydrolysate studies, among the 58 strains, Streptomyces sp. and Actinomadura sp. were the predominant genera. The other antibiotic active strains were Micromonospora sp., Microbispora sp., Nocardia sp., Pseudonocardia sp., Saccharomonospora sp., Streptoalloteichus sp. and Streptoverticillium sp. Most of them could inhibit gram-positive bacteria, especially M. luteus TISTR 884, and 8 strains (4 strains of Actinomadura, 2 strains of Micromonospora, 1 strain of Microbispora, and 1 strain of Streptomyces could inhibit both gram-positive and gram-negative bacteria.

  15. Actinomycetes: A Repertory of Green Catalysts with a Potential Revenue Resource

    Directory of Open Access Journals (Sweden)

    Divya Prakash

    2013-01-01

    Full Text Available Biocatalysis, one of the oldest technologies, is becoming a favorable alternative to chemical processes and a vital part of green technology. It is an important revenue generating industry due to a global market projected at $7 billion in 2013 with a growth of 6.7% for enzymes alone. Some microbes are important sources of enzymes and are preferred over sources of plant and animal origin. As a result, more than 50% of the industrial enzymes are obtained from bacteria. The constant search for novel enzymes with robust characteristics has led to improvisations in the industrial processes, which is the key for profit growth. Actinomycetes constitute a significant component of the microbial population in most soils and can produce extracellular enzymes which can decompose various materials. Their enzymes are more attractive than enzymes from other sources because of their high stability and unusual substrate specificity. Actinomycetes found in extreme habitats produce novel enzymes with huge commercial potential. This review attempts to highlight the global importance of enzymes and extends to signify actinomycetes as promising harbingers of green technology.

  16. Anticancer property of sediment actinomycetes against MCF-7 and MDA-MB-231 cell lines

    Institute of Scientific and Technical Information of China (English)

    Ravikumar S; Fredimoses M; Gnanadesigan M

    2012-01-01

    Objective: To investigate the anticancer property of marine sediment actinomycetes against two different breast cancer cell lines. Methods:In vitro anticancer activity was carried out against breast (MCF-7 and MDA-MB-231) cancer cell lines. Partial sequences of the 16s rRNA gene, phylogenetic tree construction, multiple sequence analysis and secondary structure analysis were also carried out with the actinomycetes isolates. Results: Of the selected five actinomycete isolates, ACT01 and ACT02 showed the IC50 value with (10.13±0.92) and (22.34±5.82)μg/mL concentrations, respectively for MCF-7 cell line at 48 h, but ACT01 showed the minimum (18.54±2.49 μg/mL) level of IC50 value with MDA-MB-231 cell line. Further, the 16s rRNA partial sequences of ACT01, ACT02, ACT03, ACT04 and ACT05 isolates were also deposited in NCBI data bank with the accession numbers of GQ478246, GQ478247, GQ478248, GQ478249 and GQ478250, respectively. The phylogenetic tree analysis showed that, the isolates of ACT02 and ACT03 were represented in group I and III, respectively, but ACT01 and ACT02 were represented in group II. The multiple sequence alignment of the actinomycete isolates showed that, the maximum identical conserved regions were identified with the nucleotide regions of 125 to 221st base pairs, 65 to 119th base pairs and 55, 48 and 31st base pairs. Secondary structure prediction of the 16s rRNA showed that, the maximum free energy was consumed with ACT03 isolate (-45.4 kkal/mol) and the minimum free energy was consumed with ACT04 isolate (-57.6 kkal/mol). Conclusions:The actinomycete isolates of ACT01 and ACT02 (GQ478246 and GQ478247) which are isolated from sediment sample can be further used as anticancer agents against breast cancer cell lines.

  17. Isolation and screening of antibiotic producing actinomycetes from soils in Gondar town, North West Ethiopia

    Directory of Open Access Journals (Sweden)

    Abebe Bizuye

    2013-10-01

    Full Text Available Objective: To isolate and screen antibiotic producing actinomycetes from potential soil samples of Gondar town, Ethiopia. Methods: Fifteen soil samples were collected, serially diluted and spread on starch casein and oat meal agar supplemented with amoxicillin and cyclohexamide for inhibition of bacteria and fungi, respectively. Cross streak method was used to check antagonistic activity of isolated actinomycetes against test organisms. Solid state fermentation and crude extraction were used for the production of antibiotics from isolates. Agar well diffusion was used for antimicrobial activity of crude extracts against test organisms. Results: Three isolates (Ab18, Ab28 and Ab43 have been shown high antagonistic activity during primary screening. Inhibition zones obtained from crude extracts showed significance differences when compared with standard antibiotics tested against test organisms (P<0.05. Inhibition zone of crude extracts from isolate Ab18 against Klebsiella pneumonia ATCC7000603 and Escherichia coli ATCC25922 were (14依1 mm and (35依1 mm, respectively which were strong active when compared to amoxicillin (0 mm and tetracycline [(13依1 mm for Klebsiella pneumonia ATCC7000603 and (33依 1 mm for Escherichia coli ATCC25922]. Crude extracts from isolate Ab18 showed (20依1 mm and (15 依1 mm inhibition zones against methicillin resistant Staphylococcus aureus strains 2 (MRSA2 and MRSA4, respectively. Crude extract from isolate Ab43 has shown inhibition zones of (16依1 mm and (17依1 mm against MRSA2 and MRSA4, respectively. Combination of Ab18 and Ab43 has shown high antimicrobial activity (18依1 mm against MRSA2 and MRSA4. Conclusions: There was not any scientific report on soil actinomycetes producing antibiotic in the study areas. Therefore, isolation and screening of actinomycetes from such areas in optimum condition may contribute the discovery of new antibiotics. Potent antibiotics from these actinomycetes could contribute a

  18. The biodegradation of layered silicates under the influence of cyanobacterial-actinomycetes associations

    Science.gov (United States)

    Ivanova, Ekaterina

    2013-04-01

    The weathering of sheet silicates is well known to be related to local and global geochemical cycles. Content and composition of clay minerals in soil determine the sorption properties of the soil horizons, water-holding capacity of the soil, stickiness, plasticity, etc. Microorganisms have a diverse range of mechanisms of minerals' structure transformation (acid- and alkali formation, biosorption, complexing, etc). One of the methods is an ability of exopolysaccharide-formation, in particular the formation of mucus, common to many bacteria, including cyanobacteria. Mucous covers cyanobacteria are the specific econiches for other bacteria, including actinomycetes. The objective was to analyze the structural changes of clay minerals under the influence of the cyanobacterial-actinomycetes associative growth. The objects of the study were: 1) the experimental symbiotic association, consisting of free-living heterocyst-formative cyanobacterium Anabaena variabilis Kutz. ATCC 294132 and actinomycete Streptomyces cyaneofuscatus FR837630, 2) rock samples obtained from the Museum of the Soil Science Department of the Lomonosov Moscow State University: kaolinite, consisting of kaolin (96%) Al4 (OH) 8 [Si4O10]; mixed with hydromica, chlorite and quartz; vermiculite, consisting of vermiculite (Ca, Mg, ...)*(Mg, Fe)3(OH)2[(Si, Al)4O10]*4H2O and trioctahedral mica (biotite). The mineralogical compositions of the rocks were determined by the universal X-ray Diffractometer Carl Zeiss Yena. The operationg regime was kept constant (30 kv, 40 mA). The cultivation of the association of actinomycete S. cyanoefuscatus and cyanobacterium A. variabilis caused a reduction in the intensity of kaolinite and hydromica reflexes. However, since both (mica and kaolinite) components have a rigid structure, the significant structural transformation of the minerals was not revealed. Another pattern was observed in the experiment, where the rock sample of vermiculite was used as the mineral

  19. Microbial Consortium with High Cellulolytic Activity (MCHCA) for Enhanced Biogas Production.

    Science.gov (United States)

    Poszytek, Krzysztof; Ciezkowska, Martyna; Sklodowska, Aleksandra; Drewniak, Lukasz

    2016-01-01

    The use of lignocellulosic biomass as a substrate in agricultural biogas plants is very popular and yields good results. However, the efficiency of anaerobic digestion, and thus biogas production, is not always satisfactory due to the slow or incomplete degradation (hydrolysis) of plant matter. To enhance the solubilization of the lignocellulosic biomass various physical, chemical and biological pretreatment methods are used. The aim of this study was to select and characterize cellulose-degrading bacteria, and to construct a microbial consortium, dedicated for degradation of maize silage and enhancing biogas production from this substrate. Over 100 strains of cellulose-degrading bacteria were isolated from: sewage sludge, hydrolyzer from an agricultural biogas plant, cattle slurry and manure. After physiological characterization of the isolates, 16 strains (representatives of Bacillus, Providencia, and Ochrobactrum genera) were chosen for the construction of a Microbial Consortium with High Cellulolytic Activity, called MCHCA. The selected strains had a high endoglucanase activity (exceeding 0.21 IU/mL CMCase activity) and a wide range of tolerance to various physical and chemical conditions. Lab-scale simulation of biogas production using the selected strains for degradation of maize silage was carried out in a two-bioreactor system, similar to those used in agricultural biogas plants. The obtained results showed that the constructed MCHCA consortium is capable of efficient hydrolysis of maize silage, and increases biogas production by even 38%, depending on the inoculum used for methane fermentation. The results in this work indicate that the mesophilic MCHCA has a great potential for application on industrial scale in agricultural biogas plants.

  20. The complete genome sequence of Fibrobacter succinogenes S85 reveals a cellulolytic and metabolic specialist.

    Directory of Open Access Journals (Sweden)

    Garret Suen

    Full Text Available Fibrobacter succinogenes is an important member of the rumen microbial community that converts plant biomass into nutrients usable by its host. This bacterium, which is also one of only two cultivated species in its phylum, is an efficient and prolific degrader of cellulose. Specifically, it has a particularly high activity against crystalline cellulose that requires close physical contact with this substrate. However, unlike other known cellulolytic microbes, it does not degrade cellulose using a cellulosome or by producing high extracellular titers of cellulase enzymes. To better understand the biology of F. succinogenes, we sequenced the genome of the type strain S85 to completion. A total of 3,085 open reading frames were predicted from its 3.84 Mbp genome. Analysis of sequences predicted to encode for carbohydrate-degrading enzymes revealed an unusually high number of genes that were classified into 49 different families of glycoside hydrolases, carbohydrate binding modules (CBMs, carbohydrate esterases, and polysaccharide lyases. Of the 31 identified cellulases, none contain CBMs in families 1, 2, and 3, typically associated with crystalline cellulose degradation. Polysaccharide hydrolysis and utilization assays showed that F. succinogenes was able to hydrolyze a number of polysaccharides, but could only utilize the hydrolytic products of cellulose. This suggests that F. succinogenes uses its array of hemicellulose-degrading enzymes to remove hemicelluloses to gain access to cellulose. This is reflected in its genome, as F. succinogenes lacks many of the genes necessary to transport and metabolize the hydrolytic products of non-cellulose polysaccharides. The F. succinogenes genome reveals a bacterium that specializes in cellulose as its sole energy source, and provides insight into a novel strategy for cellulose degradation.

  1. Interactions between Cellulolytic Enzymes with Native, Autohydrolysis, and Technical Lignins and the Effect of a Polysorbate Amphiphile in Reducing Nonproductive Binding.

    Science.gov (United States)

    Fritz, Consuelo; Ferrer, Ana; Salas, Carlos; Jameel, Hasan; Rojas, Orlando J

    2015-12-14

    Understanding enzyme-substrate interactions is critical in designing strategies for bioconversion of lignocellulosic biomass. In this study we monitored molecular events, in situ and in real time, including the adsorption and desorption of cellulolytic enzymes on lignins and cellulose, by using quartz crystal microgravimetry and surface plasmon resonance. The effect of a nonionic surface active molecule was also elucidated. Three lignin substrates relevant to the sugar platform in biorefinery efforts were considered, namely, hardwood autohydrolysis cellulolytic (HWAH), hardwood native cellulolytic (MPCEL), and nonwood native cellulolytic (WSCEL) lignin. In addition, Kraft lignins derived from softwoods (SWK) and hardwoods (HWK) were used as references. The results indicated a high affinity between the lignins with both, monocomponent and multicomponent enzymes. More importantly, the addition of nonionic surfactants at concentrations above their critical micelle concentration reduced remarkably (by over 90%) the nonproductive interactions between the cellulolytic enzymes and the lignins. This effect was hypothesized to be a consequence of the balance of hydrophobic and hydrogen bonding interactions. Moreover, the reduction of surface roughness and increased wettability of lignin surfaces upon surfactant treatment contributed to a lower affinity with the enzymes. Conformational changes of cellulases were observed upon their adsorption on lignin carrying preadsorbed surfactant. Weak electrostatic interactions were determined in aqueous media at pH between 4.8 and 5.5 for the native cellulolytic lignins (MPCEL and WSCEL), whereby a ∼20% reduction in the enzyme affinity was observed. This was mainly explained by electrostatic interactions (osmotic pressure effects) between charged lignins and cellulases. Noteworthy, adsorption of nonionic surfactants onto cellulose, in the form cellulose nanofibrils, did not affect its hydrolytic conversion. Overall, our results

  2. Succession of Actinomycetes During Composting Proccess of Dairy-Farm Waste Investigated by Culture-Dependent and Independent Approaches

    Directory of Open Access Journals (Sweden)

    Mukhlissul Faatih1

    2015-11-01

    Full Text Available Mesophilic, thermophilic, and maturation phases were recognized in composting proccess. Temperaturechanges influence the microbial communities in compost within composting proccess. Actinomycetes account for alarger part of compost microbial population. The aim of this research was to study succession of actinomycetescommunity during composting of dairy-farm waste investigated by culture-dependent and independentapproaches.In culture-independent method, the succession of actinomycetes community was analyzed by nestedpolymerasechain reaction of ribosomal intergenic spacer (nested-PCR RISA using spesific primer F243 and primerR23S followed by a second PCR using primers F968 and R23S. In culture-dependent method actinomycetes fromcompost were isolated on selective media, starch-nitrate medium and humic-acid + vitamins medium. DNA ofactinomycetes was extracted and amplified by repetitive sequence-based PCR (rep-PCR using primer BOXA1R. Thebanding patterns were used to generate dendrograms by UPGMA clustering with NTSYS program. Microcosmcontaining sterile rice-straw and water which is inoculated with each actinomycetes isolates was used for examiningthe ability of each isolate in rice-straw degradation.The experiment results showed that succession of both bacteria and actinomycetes was occured withincomposting proccess of dairy-farm waste. Analysed by culture-independent method revealed that the highestcommunity of compost’s bacteria was on mesophilic, thermophilic, and maturation phases, respectively. WhereasPCR-nested RISA resulted the highest population of actinomycetes was on thermophilic, maturation, and mesophilicphases, respectively. By culture-dependent method was obtained 29 actinomycetes isolates from mesophilic phase,23 isolates from thermophilic phase, and 19 isolates from maturation phase. Genetic diversity analysis of the obtainedisolates showed the presence of phylogenetic grouping on each phase of composting proccess. This result

  3. Functional Gene-Guided Discovery of Type II Polyketides from Culturable Actinomycetes Associated with Soft Coral Scleronephthya sp

    Science.gov (United States)

    Sun, Wei; Peng, Chongsheng; Zhao, Yunyu; Li, Zhiyong

    2012-01-01

    Compared with the actinomycetes in stone corals, the phylogenetic diversity of soft coral-associated culturable actinomycetes is essentially unexplored. Meanwhile, the knowledge of the natural products from coral-associated actinomycetes is very limited. In this study, thirty-two strains were isolated from the tissue of the soft coral Scleronephthya sp. in the East China Sea, which were grouped into eight genera by 16S rDNA phylogenetic analysis: Micromonospora, Gordonia, Mycobacterium, Nocardioides, Streptomyces, Cellulomonas, Dietzia and Rhodococcus. 6 Micromonospora strains and 4 Streptomyces strains were found to be with the potential for producing aromatic polyketides based on the analysis of KSα (ketoacyl-synthase) gene in the PKS II (type II polyketides synthase) gene cluster. Among the 6 Micromonospora strains, angucycline cyclase gene was amplified in 2 strains (A5-1 and A6-2), suggesting their potential in synthesizing angucyclines e.g. jadomycin. Under the guidance of functional gene prediction, one jadomycin B analogue (7b, 13-dihydro-7-O-methyl jadomycin B) was detected in the fermentation broth of Micromonospora sp. strain A5-1. This study highlights the phylogenetically diverse culturable actinomycetes associated with the tissue of soft coral Scleronephthya sp. and the potential of coral-derived actinomycetes especially Micromonospora in producing aromatic polyketides. PMID:22880121

  4. SELEKSI DAN PEMANFAATAN ACTINOMYCETES SEBAGAI MIKROBA ANTAGONIS YANG RAMAH LINGKUNGAN TERHADAP Fusarium oxysporum f.sp. cubense SECARA IN VITRO

    Directory of Open Access Journals (Sweden)

    I MADE SUDARMA

    2015-06-01

    Full Text Available A total of 119 different actinomycete isolate were recovered from banana crop habitats with and without Fusarium wilt disease symptom. These were than assessed for their antagonist ability against Fusarium oxysporum £sp. cubense (Foe in vitro. Results indicated that four of all actinomycete isolate active against Foe. The four of actinomycete isolates were Streptomyces sp. l (AAo4, Streptomyces sp.2 (AAo32 , Streptomyces sp.3 (AAo33 and Streptomyces sp. 4 (AAo35. It was can inhibit the Foe mycelium growth, 79,63%, 72,22%, 78,89% and 72,22% respectively. After tested with the 3 times replication, the four Streptomyces spp. isolate effective to control the Foe that attack Bali banana cultivars, such as Susu, Saba, Raja and Ketip.

  5. Isolation of aerobic cultivable cellulolytic bacteria from different regions of the gastrointestinal tract of giant land snail Achatina fulica

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    Guilherme L. Pinheiro

    2015-08-01

    Full Text Available The enzymatic hydrolysis of cellulose by cellulases is one of the major limiting steps in the conversion of lignocellulosic biomass to yield bioethanol. To overcome this hindrance, significant efforts are underway to identify novel cellulases. The snail Achatina fulica is a gastropod with high cellulolytic activity, mainly due to the abundance of glycoside hydrolases produced by both the animal and its resident microbiota. In this study, we partially assessed the cellulolytic bacterial diversity inside the gastrointestinal tract of A. fulica by culture-dependent methods and evaluated the hydrolytic repertoire of the isolates. Forty bacterial isolates were recovered from distinct segments of the snail gut and identified to the genus level by 16S rRNA gene sequence analysis. Additional phenotypic characterization was performed using biochemical tests provided by the Vitek2 identification system. The overall enzymatic repertoire of the isolated strains was investigated by enzymatic plate assays, containing the following substrates: powdered sugarcane bagasse, carboxymethylcellulose (CMC, p-nitrophenyl-b-D-glucopyranoside (pNPG, p-nitrophenyl-b-D-cellobioside (pNPC, 4-methylumbelliferyl-b-D-glucopyranoside (MUG, 4-methylumbelliferyl-b-D-cellobioside (MUC and 4-methylumbelliferyl-b-D-xylopyranoside (MUX. Our results indicate that the snail Achatina fulica is an attractive source of cultivable bacteria that showed to be valuable resources for the production of different types of biomass-degrading enzymes.

  6. Comparative genotyping of Clostridium thermocellum strains isolated from biogas plants: genetic markers and characterization of cellulolytic potential.

    Science.gov (United States)

    Koeck, Daniela E; Zverlov, Vladimir V; Liebl, Wolfgang; Schwarz, Wolfgang H

    2014-07-01

    Clostridium thermocellum is among the most prevalent of known anaerobic cellulolytic bacteria. In this study, genetic and phenotypic variations among C. thermocellum strains isolated from different biogas plants were determined and different genotyping methods were evaluated on these isolates. At least two C. thermocellum strains were isolated independently from each of nine different biogas plants via enrichment on cellulose. Various DNA-based genotyping methods such as ribotyping, RAPD (Random Amplified Polymorphic DNA) and VNTR (Variable Number of Tandem Repeats) were applied to these isolates. One novel approach - the amplification of unknown target sequences between copies of a previously discovered Random Inserted Mobile Element (RIME) - was also tested. The genotyping method with the highest discriminatory power was found to be the amplification of the sequences between the insertion elements, where isolates from each biogas plant yielded a different band pattern. Cellulolytic potentials, optimal growth conditions and substrate spectra of all isolates were characterized to help identify phenotypic variations. Irrespective of the genotyping method used, the isolates from each individual biogas plant always exhibited identical patterns. This is suggestive of a single C. thermocellum strain exhibiting dominance in each biogas plant. The genotypic groups reflect the results of the physiological characterization of the isolates like substrate diversity and cellulase activity. Conversely, strains isolated across a range of biogas plants differed in their genotyping results and physiological properties. Both strains isolated from one biogas plant had the best specific cellulose-degrading properties and might therefore achieve superior substrate utilization yields in biogas fermenters.

  7. Temporal variations in microbial biomass C and cellulolytic enzyme activity in arable soils: effects of organic matter input

    DEFF Research Database (Denmark)

    Debosz, K.; Rasmussen, Peter Have; Pedersen, A. R.

    1999-01-01

    Temporal variations in soil microbial biomass C concentration and in activity of extracellular enzymes of the cellulolytic complex were investigated in a field experiment after eight years of cultivation with either low organic matter input (low-OM) or high organic matter input (high-OM). The cul......Temporal variations in soil microbial biomass C concentration and in activity of extracellular enzymes of the cellulolytic complex were investigated in a field experiment after eight years of cultivation with either low organic matter input (low-OM) or high organic matter input (high......-OM). The cultivation systems differed in whether their source of fertiliser was mainly mineral or organic, in whether a winter cover crop was grown, and whether straw was mulched or removed. Sampling occurred at approximately monthly intervals, over a period of two years. Distinct temporal variations in microbial......) and an endocellulase activity of 44.2 +/- 1.1 nmol g(-1) h(-1). (C) 1999 Elsevier Science B.V. All rights reserved....

  8. Characterization of cellulolytic enzymes and bioH2 production from anaerobic thermophilic Clostridium sp. TCW1.

    Science.gov (United States)

    Lo, Yung-Chung; Huang, Chi-Yu; Cheng, Chieh-Lun; Lin, Chiu-Yue; Chang, Jo-Shu

    2011-09-01

    A thermophilic anaerobic bacterium Clostridium sp. TCW1 was isolated from dairy cow dung and was used to produce hydrogen from cellulosic feedstock. Extracellular cellulolytic enzymes produced from TCW1 strain were identified as endoglucanases (45, 53 and 70 kDa), exoglucanase (70 kDa), xylanases (53 and 60 kDa), and β-glucosidase (45 kDa). The endoglucanase and xylanase were more abundant. The optimal conditions for H2 production and enzyme production of the TCW1 strain were the same (60 °C, initial pH 7, agitation rate of 200 rpm). Ten cellulosic feedstock, including pure or natural cellulosic materials, were used as feedstock for hydrogen production by Clostridium strain TCW1 under optimal culture conditions. Using filter paper at 5.0 g/L resulted in the most effective hydrogen production performance, achieving a H2 production rate and yield of 57.7 ml/h/L and 2.03 mol H2/mol hexose, respectively. Production of cellulolytic enzyme activities was positively correlated with the efficiency of dark-H2 fermentation.

  9. Direct ethanol production from cellulosic materials at high temperature using the thermotolerant yeast Kluyveromyces marxianus displaying cellulolytic enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Yanase, Shuhei; Yamada, Ryosuke; Ogino, Chiaki; Kondo, Akihiko [Kobe Univ. (Japan). Dept. of Chemical Science and Engineering; Hasunuma, Tomohisa; Tanaka, Tsutomu; Fukuda, Hideki [Kobe Univ. (Japan). Organization of Advanced Science and Technology

    2010-09-15

    To exploit cellulosic materials for fuel ethanol production, a microorganism capable of high temperature and simultaneous saccharification-fermentation has been required. However, a major drawback is the optimum temperature for the saccharification and fermentation. Most ethanol-fermenting microbes have an optimum temperature for ethanol fermentation ranging between 28 C and 37 C, while the activity of cellulolytic enzymes is highest at around 50 C and significantly decreases with a decrease in temperature. Therefore, in the present study, a thermotolerant yeast, Kluyveromyces marxianus, which has high growth and fermentation at elevated temperatures, was used as a producer of ethanol from cellulose. The strain was genetically engineered to display Trichoderma reesei endoglucanase and Aspergillus aculeatus {beta}-glucosidase on the cell surface, which successfully converts a cellulosic {beta}-glucan to ethanol directly at 48 C with a yield of 4.24 g/l from 10 g/l within 12 h. The yield (in grams of ethanol produced per gram of {beta}-glucan consumed) was 0.47 g/g, which corresponds to 92.2% of the theoretical yield. This indicates that high-temperature cellulose fermentation to ethanol can be efficiently accomplished using a recombinant K. marxianus strain displaying thermostable cellulolytic enzymes on the cell surface. (orig.)

  10. Isolation of aerobic cultivable cellulolytic bacteria from different regions of the gastrointestinal tract of giant land snail Achatina fulica

    Science.gov (United States)

    Pinheiro, Guilherme L.; Correa, Raquel F.; Cunha, Raquel S.; Cardoso, Alexander M.; Chaia, Catia; Clementino, Maysa M.; Garcia, Eloi S.; de Souza, Wanderley; Frasés, Susana

    2015-01-01

    The enzymatic hydrolysis of cellulose by cellulases is one of the major limiting steps in the conversion of lignocellulosic biomass to yield bioethanol. To overcome this hindrance, significant efforts are underway to identify novel cellulases. The snail Achatina fulica is a gastropod with high cellulolytic activity, mainly due to the abundance of glycoside hydrolases produced by both the animal and its resident microbiota. In this study, we partially assessed the cellulolytic aerobic bacterial diversity inside the gastrointestinal tract of A. fulica by culture-dependent methods and evaluated the hydrolytic repertoire of the isolates. Forty bacterial isolates were recovered from distinct segments of the snail gut and identified to the genus level by 16S rRNA gene sequence analysis. Additional phenotypic characterization was performed using biochemical tests provided by the Vitek2 identification system. The overall enzymatic repertoire of the isolated strains was investigated by enzymatic plate assays, containing the following substrates: powdered sugarcane bagasse, carboxymethylcellulose (CMC), p-nitrophenyl-β-D-glucopyranoside (pNPG), p-nitrophenyl-β-D-cellobioside (pNPC), 4-methylumbelliferyl-β-D-glucopyranoside (MUG), 4-methylumbelliferyl-β-D-cellobioside (MUC), and 4-methylumbelliferyl-β-D-xylopyranoside (MUX). Our results indicate that the snail A. fulica is an attractive source of cultivable bacteria that showed to be valuable resources for the production of different types of biomass-degrading enzymes. PMID:26347735

  11. Degradation of cellulose by the bean-pathogenic fungus Colletotrichum lindemuthianum. Production of extracellular cellulolytic enzymes by cellulose induction.

    Science.gov (United States)

    Acosta-Rodríguez, Ismael; Piñón-Escobedo, Carlos; Zavala-Páramo, Ma Guadalupe; López-Romero, Everardo; Cano-Camacho, Horacio

    2005-05-01

    Colletotrichum lindemuthianum was able to grow and produce extracellular cellulolytic activity in a defined medium containing cellulose as the main carbon substrate. As measured either by the hydrolysis of 4-methylumbelliferyl-beta-D -cellotrioside or the release of glucose from carboxymethylcellulose, activity reached a peak after 13 days of incubation and then declined whereas growth markedly increased afterwards. Detection of glucose in carboxymethylcellulose hydrolysates suggested the concerted operation of endo-1,4-beta-glucanase, cellobiohydrolase (exo-1,4-beta-glucanase) and beta-glucosidase activities. The highest levels of cellulolytic activity were obtained in media supplemented with cellulose and glutamate. Other carbon and nitrogen sources markedly influenced growth and enzyme production. Oligonucleotides homologous to specific regions of the cellobiohydrolase-encoding cbhII gene from Trichoderma reesei were used to isolate a C. lindemuthianum cbhII-DNA fragment whose sequence revealed homologies of 98% and 92% with the nucleotide and the deduced amino acid sequences of the corresponding cbhII-DNA of T. reesei, respectively. RT-PCR and Southern blot analyses of total RNA samples obtained from cellulose-grown but not from glucose-grown mycelium revealed the expression of the corresponding cbhII transcript. The cbhII-cDNA fragment was cloned and sequenced.

  12. THE INFLUENCE OF KAPOK (Ceiba pentandra SEED OIL SUPPLEMENTATION ON CELLULOLYTIC ENZYME AND RUMEN MICROBIAL FERMENTATION ACTIVITY OF LOCAL SHEEP

    Directory of Open Access Journals (Sweden)

    W. Widiyanto

    2014-10-01

    Full Text Available This research was conducted to study the influence of kapok seed oil (KSO supplementation oncellulolytic enzyme and microbial fermentation activity. Sheep rumen fluid was used as enzyme sourceand inoculant, whereas carboxymethylcellulose (CMC was used as the substrate. There were 4 levels ofKSO supplementation as treatment, i.e. : 0% (T0, 5% (T1, 10% (T2, and 15% (T3. Two measuredvariables were reduced sugar production rate and gas fermentation production. The data were analyzedby analysis of variance in completely randomized design. The result showed that reduced sugarproduction rate in T0, T1, T2 and T3 treatment groups were 2.58; 2.93; 2.08 and 1.58 mg/gCMC/minute, respectively, whereas gas production were : 15.97; 13.26; 10.54 and 7.57 mg/g CMC,respectively. Kapok seed oil supplementation up to 5% DM of cellulose substrate (CMC did notinfluence the ruminal cellulolytic enzyme activity. The KSO supplementation level 10% - 15%decreased the ruminal cellulolytic enzyme activity.

  13. Purification and characterization of protease enzyme from actinomycetes and its cytotoxic effect on cancer cell line (A549)

    Institute of Scientific and Technical Information of China (English)

    C Balachandran; V Duraipandiyan; S Ignacimuthu

    2012-01-01

    Objective: To isolate active actinomycetes from soil samples of Northern Himalayas and study their culture characterization, protease production and cytotoxic effects on cancer cell line (A549). Methods: Forty six strains of actinomycetes were isolated from the soil collected from Northern Himalayas, India. Isolation of actinomycetes was performed by serial dilution plate technique. Forty six isolated actinomycetes cultures were grown in ISP 2 medium to study the morphology and biochemical characteristics. Isolated strains were studied for protease enzyme production in skim milk agar medium with solubilising capacity. Seven isolates were studied for melanin pigmentation and different NaCl concentration. Effects of environmental conditions influencing protease enzyme production of seven isolated strains were also studied at different pH, temperature and metal ions (β-mercaptoethanol, dithiothreitol, iodoacetamide, MgSO4, CaCl2 and EDTA). The seven isolates were also studied for lytic enzyme activity using different bacteria and yeast such as Pseudomonas aeruginosa (P. aeruginosa), Enterococcus feacalis (E. feacalis), Escherishia coli (E. coli), Candida albicans (C. albicans), Bacillus subtilis (B. subtilis), Klebsiella pneumonia (K. pneumonia) and Staphylococcus aureus (S. aureus). Results: Isolates ERIA-31 and ERIA-33 produced more protease enzyme activity in modified nutrient agar media compared to other actinomycetes cultures. ERIA-31 and ERIA-33 were tested for cytotoxic effect in human adenocarcinoma cancer cell line (A549). IC50 for ERIA-31 was 57.04 μg/mL and IC50 for ERIA-33 was 55.07 μg/mL. Conclusion: Actinomycete being a protease producing bacteria has the potential for use in industrial purpose, pharmaceuticals, cytotoxic agent and its proteolytic activity. Isolates of ERIA-31 and ERIA-33 produced significant amount of protease enzymes.

  14. Isolation and characterization of potential antibiotic producing actinomycetes from water and sediments of Lake Tana, Ethiopia

    Institute of Scientific and Technical Information of China (English)

    Gebreselema Gebreyohannes; Feleke Moges; Samuel Sahile; Nagappan Raja

    2013-01-01

    To isolate, evaluate and characterize potential antibiotic producing actinomycetes from water and sediments of Lake Tana, Ethiopia. Methods: A total of 31 strains of actinomycetes were isolated and tested against Gram positive and Gram negative bacterial strains by primary screening. In the primary screening, 11 promising isolates were identified and subjected to solid state and submerged state fermentation methods to produce crude extracts. The fermented biomass was extracted by organic solvent extraction method and tested against bacterial strains by disc and agar well diffusion methods. The isolates were characterized by using morphological, physiological and biochemical methods. Results: The result obtained from agar well diffusion method was better than disc diffusion method. The crude extract showed higher inhibition zone against Gram positive bacteria than Gram negative bacteria. One-way analysis of variance confirmed most of the crude extracts were statistically significant at 95% confidence interval. The minimum inhibitory concentration and minimum bactericidal concentration of crude extracts were 1.65 mg/mL and 3.30 mg/mL against Staphylococcus aureus, and 1.84 mg/mL and 3.80 mg/mL against Escherichia coli respectively. The growth of aerial and substrate mycelium varied in different culture media used. Most of the isolates were able to hydrolysis starch and urea; able to survive at 5% concentration of sodium chloride; optimum temperature for their growth was 30 °C. Conclusions: The results of the present study revealed that freshwater actinomycetes of Lake Tana appear to have immense potential as a source of antibacterial compounds.

  15. Marine Actinomycetes screening of Banten West Coast and their antibiotics purification

    Directory of Open Access Journals (Sweden)

    ROFIQ SUNARYANTO

    2010-10-01

    Full Text Available Sunaryanto R, Marwoto B (2010 Marine Actinomycetes screening of Banten West Coast and their antibiotics purification. Biodiversitas 11: 176-181. Isolation and purification of active compounds produced by marine Actinomycetes has been carried out. Marine sediment samples were obtained from six different places at Anyer, Banten West Coast in October 20, 2007. Isolation was carried out using two methods pretreatments, acid treatment and heat shock treatment. A total of 29 Actinomycetes isolates were obtained from the various sediment samples collected, then tested for antimicrobial test against Escherichia coli ATCC 25922, Staphylococcus aureus ATCC25923, Pseudomonas aeruginosa ATCC27853, Bacillus subtilis ATCC 66923, Candida albicans BIOMCC00122 and Aspergillus niger BIOMCC00134. Identification of potential isolate was carried out using 16S rRNA. Purification of active compound was carried out using silica gel column chromatography and preparative HPLC. Result of this research showed that isolate A11 produced the most active compound against Gram-positive and Gram-negative bacteria. Morphology and identification test using 16S rRNA gen showed that isolate A11 is Streptomyces sp. Production of active compound from isolate A11 used yeast peptone medium. The single peak of active compound was detected by HPLC and showed retention time on 8.35 min and maximum absorbance UV visible of antibiotic was 210 nm and 274.5 nm. Active purified compound showed inhibition activity to Gram-positive and Gram-negative bacteria. Minimum inhibitory concentration (MIC to E. coli ATCC 25922 was 27 µg/mL, P. aeruginosa ATCC 27853 68.7 µg/mL, S. aureus ATCC 25923 80.2 µg/mL, and B. subtilis ATCC 66923 73.7 µg/mL.

  16. Lignin-solubilizing ability of actinomycetes isolated from termite (Termitidae) gut.

    Science.gov (United States)

    Pasti, M B; Pometto, A L; Nuti, M P; Crawford, D L

    1990-01-01

    The lignocellulose-degrading abilities of 11 novel actinomycete strains isolated from termite gut were determined and compared with that of the well-characterized actinomycete, Streptomyces viridosporus T7A. Lignocellulose bioconversion was followed by (i) monitoring the degradation of [14C]lignin- and [14C]cellulose-labeled phloem of Abies concolor to 14CO2 and 14C-labeled water-soluble products, (ii) determining lignocellulose, lignin, and carbohydrate losses resulting from growth on a lignocellulose substrate prepared from corn stalks (Zea mays), and (iii) quantifying production of a water-soluble lignin degradation intermediate (acid-precipitable polymeric lignin). The actinomycetes were all Streptomyces strains and could be placed into three groups, including a group of five strains that appear superior to S. viridosporus T7A in lignocellulose-degrading ability, three strains of approximately equal ability, and three strains of lesser ability. Strain A2 was clearly the superior and most effective lignocellulose decomposer of those tested. Of the assays used, total lignocellulose weight loss was most useful in determining overall bioconversion ability but not in identifying the best lignin-solubilizing strains. A screening procedure based on 14CO2 evolution from [14C-lignin]lignocellulose combined with measurement of acid-precipitable polymeric lignin yield was the most effective in identifying lignin-solubilizing strains. For the termite gut strains, the pH of the medium showed no increase after 3 weeks of growth on lignocellulose. This is markedly different from the pattern observed with S. viridosporus T7A, which raises the medium pH considerably. Production of extracellular peroxidases by the 11 strains and S. viridosporus T7A was followed for 5 days in liquid cultures.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2167628

  17. Screening of actinomycetes from earthworm castings for their antimicrobial activity and industrial enzymes

    Directory of Open Access Journals (Sweden)

    Vijay Kumar

    2012-03-01

    Full Text Available Actinomycetes from earthworm castings were isolated and screened for their antimicrobial activity and industrial enzymes. A total of 48 isolates were obtained from 12 samples of earthworm castings. Highest numbers of isolates were recovered from forest site (58.33 % as compared to grassland (25% and agricultural land (16.66%. The growth patterns, mycelial coloration of abundance actinomycetes were documented. The dominant genera Identified by cultural, morphological and physiological characteristics were Streptomyces (60.41% followed by Streptosporangium (10.41%, Saccharopolyspora (6.25% and Nocardia (6.25%. Besides these, other genera like Micromonospora, Actinomadura, Microbispora, Planobispora and Nocardiopsis were also recovered but in low frequency. Among the 48 isolates, 52.08% were found active against one or more test organisms. Out of 25 active isolates 16% showed activity against bacterial, human fungal as well as phytopathogens. Among 48 isolates 38, 32, 21, 20, 16 and 14 produced enzyme amylase, caseinase, cellulase, gelatinase, xylanase and lipase respectively while 10 isolates produced all the enzymes. More interestingly 2, 3, and 1 isolates produced amylase, xylanase and lipase at 45°C respectively. In the view of its antimicrobial activity as well as enzyme production capability the genus Streptomyces was dominant. The isolate EWC 7(2 was most promising on the basis of its interesting antimicrobial activity and was identified as Streptomyces rochei. The results of these findings have increased the scope of finding industrially important actinomycetes from earthworm castings and these organisms could be promising sources for industrially important molecules or enzymes.

  18. Acetylcholinesterase inhibitory dimeric indole derivatives from the marine actinomycetes Rubrobacter radiotolerans.

    Science.gov (United States)

    Li, Jian Lin; Huang, Lei; Liu, Juan; Song, Yan; Gao, Jie; Jung, Jee H; Liu, Yonghong; Chen, Guangtong

    2015-04-01

    Investigation of the bioactive secondary metabolites of the marine actinomycetes Rubrobacter radiotolerans led to the isolation and characterization of two naturally rare dimeric indole derivatives (1 and 2). The structures of these new compounds were elucidated by spectroscopic data interpretation, and the absolute configurations were assigned by CD calculations. The acetylcholinesterase (AchE) inhibitory activity of compounds 1 and 2 was evaluated, both of which showed moderate activity with IC50 values of 11.8 and 13.5μM, respectively. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Use of bacteriophage for the selective isolation of thermophilic actinomycetes from composted eucalyptus bark.

    Science.gov (United States)

    Kurtböke, D I; Murphy, N E; Sivasithamparam, K

    1993-01-01

    A method was developed to reduce the numbers of thermophilic bacteria on isolation plates, which in turn facilitated the detection and isolation of thermophilic actinomycetes. The method involves exposing the test material to bacteriophage suspensions prior to inoculation on isolation plates. This method was applied to composted eucalyptus bark samples, which were then inoculated on R8 and 1/2 TSA + 0.2% casein hydrolysate agar plates. The phage susceptibility of thermophilic bacteria provided a selective means of reducing their numbers on isolation plates and hence increased the numbers of Thermomonospora, Saccharopolyspora rectivirgula, and thermophilic Streptomyces spp. on these media in comparison with the numbers recorded from control plates.

  20. A New Benzofuran Glycoside and Indole Alkaloids from a Sponge-Associated Rare Actinomycete, Amycolatopsis sp.

    Directory of Open Access Journals (Sweden)

    Yun Kwon

    2014-04-01

    Full Text Available Three new secondary metabolites, amycofuran (1, amycocyclopiazonic acid (2, and amycolactam (3, were isolated from the sponge-associated rare actinomycete Amycolatopsis sp. Based on combined spectroscopic analyses, the structures of 1–3 were determined to be a new benzofuran glycoside and new indole alkaloids related to cyclopiazonic acids, a class that has previously only been reported in fungi. The absolute configurations of 1 and 3 were deduced by ECD calculations, whereas that of 2 was determined using the modified Mosher method. Amycolactam (3 displayed significant cytotoxicity against the gastric cancer cell line SNU638 and the colon cancer cell line HCT116.

  1. Metabolic engineering of antibiotic factories: New tools for antibiotic production in actinomycetes

    DEFF Research Database (Denmark)

    Weber, Tilmann; Charusanti, Pep; Musiol-Kroll, Ewa Maria

    2015-01-01

    -resistant pathogens and the development of new technologies to find and produce such compounds have again attracted interest in this field. Based on improvements in whole-genome sequencing, novel methods have been developed to identify the secondary metabolite biosynthetic gene clusters by genome mining, to clone......Actinomycetes are excellent sources for novel bioactive compounds, which serve as potential drug candidates for antibiotics development. While industrial efforts to find and develop novel antimicrobials have been severely reduced during the past two decades, the increasing threat of multidrug...

  2. EXPLORATION OF ACTINOMYCETES ENDOPHYTICALLY ASSOCIATED WITH PIPER NIGRUM FOR POTENTIAL BIOACTIVITY

    Directory of Open Access Journals (Sweden)

    Jasim B.

    2015-02-01

    Full Text Available Piper nigrum is well known for its metabolite richness. So endophytic microorganisms that reside within such environments can be expected to have promising biosynthetic potential. The current study identified three endophytic actinomycetes with broad bioactivity which can have applications in natural product related pharmacological research. The Verrucosispora sp identified in the study was found to have promising anticancer and antimicrobial activities and Streptomyces sp. was found to have antioxidant activity. The results obtained are supported by many previous reports and this suggests the isolates obtained in the study to have the possible presence of potential known or novel compounds with broad spectrum of activity.

  3. Characterization of cytotoxic compound from marine sediment derived actinomycete Streptomyces avidinii strain SU4

    Institute of Scientific and Technical Information of China (English)

    Sudha S; Masilamani Selvam M

    2012-01-01

    To investigate the cytotoxic activity of actinomycete isolated from marine sediment. Methods: In the present study the DNA was isolated and the ITS region of 16s rRNA was amplified by polymerase chain reaction, using two universal bacterial primers, 1492R (5′-GGTTACCTTGTTAC GACTT-3′) and Eubac27F (5′-AGAGTTTGATCCTGGCTC AG-3′). The amplified products were purified using TIANgel mini purification kit, ligated to MD18-T simple vector (TaKaRa), and transformed into competent cells of Escherichia coli DH5α. 16S rRNA gene fragment was sequenced using forward primer M13F (-47) and reverse primer M13R (-48). Blast search sequence similarity was found against the existing non-redundant nucleotide sequence database thus, identified as Streptomyces sp SU, Streptomyces rubralavandulae strain SU1, Streptomyces cacaoi strain SU2, Streptomyces cavourensis strain SU3, Streptomyces avidinii strain SU4, Streptomyces globisporus strain SU5, Streptomyces variabilis strain SU6, Streptomycescoelicolor strain SU 7. Among the eight identified isolates, one actinomycete Streptomyces avidinii strain SU4 was selected for further study. Results: Crude extract of the actinomycete isolate exhibited IC50 in 64.5 μg against Hep-2 cell line, 250 μg in VERO cell line. This value is very close to the criteria of cytotoxicity activity for the crude extracts, as established by the American National Cancer Institute (NCI) is in IC50 < 30 μg /mL. The GC MS analysis showed that the active principle might be 1,2-benzenedicarboxylic acid, bis(2-methylpropyl) ester (12.17%), isooctyl phthalate (15.29%) with the retention time 15.642 and 21.612, respectively. Conclusions: This study clearly proves that the marine sediment derived actinomycetes with bioactive metabolites can be expected to provide high quality biological material for high throughout biochemical and anticancer screening programs. These results help us to conclude that the potential of using metabolic engineering and post genomic

  4. Characterization of cytotoxic compound from marine sediment derived actinomycete Streptomyces avidinii strain SU4

    Institute of Scientific and Technical Information of China (English)

    Sudha; S; Masilamani; Selvam; M

    2012-01-01

    Objective:To investigate the cytotoxic activity of actinomycete isolated from marine sediment.Methods:In the present study the DNA was isolated and the ITS region of 16s rRNA was amplified by polymerase chain reaction,using two universal bacterial primers,1492K(5’-GGTTACCTTG’TTAC GACTT-3’)and Eubac27F(5’-AGAGTTTGATCCTGGCTC AG-3’).The amplified products were purified using TIANgel mini purification kit,ligated to MD18-T simple vector(TaKaRa),and transformed into competent cells of Escherichia coli DH5α.16S rRNA gene fragment was sequenced using forward primer M13F(-47)and reverse primer M13R(-48).Blast search sequence similarity was found against the existing non-redundanl nucleotide sequence database thus,identified as Streptomyces sp SU,Streptomyces rubralavandulae strain SU1,Streptomyces cacaoi strain SU2,Streptomyces cavourensis strain SU3,Streptomyces avidinii strain SU4,Streptomyces globisporus strain SU5,Streptomyces variabilis strain SU6,Streptomyces coelicolor strain SU 7.Among the eight identified isolates,one actinomycete Streptomyces avidinii strain SU4 was selected for further study.Results:Crude extract of the actinomycete isolate exhibited IC50in 64.5μg against Hep-2 cell line,250μg in VERO cell line.This value is very close to the criteria of cytotoxicity activity for the crude extracts,as established by the American National Cancer Institute(NCI)is in IC50<30μg/mL.The CC MS analysis showed that the active principle might be 1,2-benzenedicarboxylic acid,bis(2-methylpropyl)ester(12.17%),isooctyl phthalate(15.29%)with the retention time 15.642 and 21.612,respectively.Conclusions:This study clearly proves that the marine sediment derived actinomycetes with bioactive metabolites can be expected to provide high quality biological material for high throughout biochemical and anticancer screening programs.These results help us to conclude that the potential of using metabolic engineering and post genomic

  5. Utilization of mixed cellulolytic microbes from termite extract, elephant faecal solution and buffalo ruminal fluid to increase in vitro digestibility of King Grass

    Directory of Open Access Journals (Sweden)

    Agung Prabowo

    2007-06-01

    Full Text Available Cellulose is a compound of plant cell walls which is difficult to be degraded because it composed of glucose monomers linked by β-(1.4-bound. It will be hydrolysed by cellulase enzyme secreted by cellulolytic microbes. The effective digestion of cellulose needs high activity of cellulase enzyme. This research aims to increase in vitro king grass digestibility utilizing mixed cellulolytic microbes of termite extract, elephant faecal solution, and buffalo ruminal fluid. Twelve syringes contained gas test media were randomly divided into four treatments based on sources of microbe (SM, namely: S (SM: cattle ruminal fluid [S], RGK (SM: mixed cellulolytic microbes of termite extract, elephant faecal solution, and buffalo ruminal fluid [RGK], with composition 1 : 1 : 1, S-RGK (SM: S + RGK, with composition 1:1, and TM (without given treatment microbe. Digestibility was measured using gas test method. Average of gas production treatment of S-RGK (70.2 + 0.6 ml was higher and significantly different (P<0.01 compared to treatment of S (60.3 + 0.8 ml, RGK (40.8 + 2.3 ml, and TM (13.3 + 2.0 ml. Utilization of mixed cellulolytic microbes of termite extract, elephant faecal solution, and buffalo ruminal fluid (RGK that combined with microbes of cattle ruminal fluid (S could increase in vitro digestibility of king grass.

  6. Screening Antimicrobial Activity of Actinomycetes Isolated from Raja Ampat, West Papua, Indonesia

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    Wellyzar Sjamsuridzal

    2012-04-01

    Full Text Available In the framework of exploitation of antimicrobial activity of Actinomycetes in Papua, one hundred isolates ofActinomycetes isolated from soil and leaf litter samples from various ecosystems in Batanta and Salawati Island, RajaAmpat, West Papua were screened. We obtained 200 crude extracts from 100 isolates based on two extraction phases.Nonpolar metabolites were extracted by ethyl acetate : methanol (4:1 solvent while the polar metabolites wereconcentrated using a freeze-drying method. Based on the agar dilution method, a total of 43 from 200(21.5% crudeextracts have antimicrobial activity against bacteria and yeasts (Escherichia coli NBRC 14237, Bacillus subtilis NBRC3134, Staphylococcus aureus NBRC 13276, Micrococcus luteus NBRC 1367, Candida albicans NBRC 1594 andSaccharomyces cerevisiae NBRC 10217. Some crude extracts showed anti-Gram negative (1.5%, anti-Gram positive(17% and antifungal (17% activities. Crude metabolites which were extracted using ethyl acetate : methanol weremore effective on antimicrobial activity (35% compared with water extraction (17%. Five most potential isolates (BL-13-5, BL-06-5, BL-14-2, BL-22-3, and Sl-36-1 were identified based on 16S rRNA gene sequence data. Sequencesimilarity search by BLAST program revealed that they show sequence similarities to Streptomyces kanamyceticus(92%, Streptomyces verne (92%, Streptomyces narbonensis (92%, Streptomyces malachitofuscus (98%, andStreptomyces hygroscopicus (96%, respectively.

  7. Genome sequencing reveals complex secondary metabolome in themarine actinomycete Salinispora tropica

    Energy Technology Data Exchange (ETDEWEB)

    Udwary, Daniel W.; Zeigler, Lisa; Asolkar, Ratnakar; Singan,Vasanth; Lapidus, Alla; Fenical, William; Jensen, Paul R.; Moore, BradleyS.

    2007-05-01

    Recent fermentation studies have identified actinomycetes ofthe marine-dwelling genus Salinispora as prolific natural productproducers. To further evaluate their biosynthetic potential, we analyzedall identifiable secondary natural product gene clusters from therecently sequenced 5,184,724 bp S. tropica CNB-440 circular genome. Ouranalysis shows that biosynthetic potential meets or exceeds that shown byprevious Streptomyces genome sequences as well as other naturalproduct-producing actinomycetes. The S. tropica genome features ninepolyketide synthase systems of every known formally classified family,non-ribosomal peptide synthetases and several hybrid clusters. While afew clusters appear to encode molecules previously identified inStreptomyces species,the majority of the 15 biosynthetic loci are novel.Specific chemical information about putative and observed natural productmolecules is presented and discussed. In addition, our bioinformaticanalysis was critical for the structure elucidation of the novelpolyenemacrolactam salinilactam A. This study demonstrates the potentialfor genomic analysis to complement and strengthen traditional naturalproduct isolation studies and firmly establishes the genus Salinispora asa rich source of novel drug-like molecules.

  8. Antimethicilin resistance agents from marine actinomycetes from soil sediments of Lagos Lagoon

    Institute of Scientific and Technical Information of China (English)

    Davies Olabisi Flora; Adeleye Isaac Adeyemi; Wang Peng George

    2015-01-01

    Objective:To evaluate theisolation of actinomycetes strains with potential for producing antimicrobials with high methicilin resistance capability. Methods: The soil samples were collected from four different locations of Lagos lagoon. TheActinomycetes were isolated from the samples by serial dilution using spread plate method. Isolates were selected based on their cultural characteristics as well as their Gram reaction and phenotypically and molecularly characterizedStreptomyces sp. Isolates were inoculated in starch casein and Kuster’s broth media and secondary metabolites were screened for antimicrobial activity against the following microorganisms: methicillin resistant Staphylococcus aureus,Staphylococcus aureusATCC29213,Escherichia coliATCC 29522, Pseudomonas aeruginosaATCC27853,Candida albicans,Enterococcus faecalisATCC 29212. Coagulase-negative staphylococci isolated fromHIV patients were also used (Staphylococcus warneri,Staphylococcus xylosus andStaphylococcus epidermidis). The antimicrobial metabolites of the isolates were identified using gas chromatography-mass spectrometer. Results:Extracts from isolatesULS12 andULS13 showed antimicrobial activity against methicillin resistantStaphylococcus aureus whileULK3 inhibitedCandida albicans only. The gas chromatography-mass spectrometerdata analysis showed the antibiotic profile of these isolates. Conclusions: The isolatesULS12 andULS13 were found to display the highest antimicrobial activity against the test organisms and could be a potential source of new antibiotics.

  9. Isolation and characterization of Cr(VI)-reducing actinomycetes from estuarine sediments.

    Science.gov (United States)

    Terahara, Takeshi; Xu, Xudan; Kobayashi, Takeshi; Imada, Chiaki

    2015-04-01

    Bioremediation technologies have strong potential use in the less costly and more environmentally friendly removal of highly toxic hexavalent-chromium (Cr(VI)) compared with physicochemical technologies. Several Cr(VI)-reducing bacteria have been isolated; however, there are few studies on Cr(VI)-resistant and Cr(VI)-reducing actinomycetes. In this study, Cr(VI)-reducing actinomycetes were screened from estuarine, marine, and terrestrial samples on the basis of Cr(VI)-resistant and Cr(VI)-reducing ability. Of the 80 Streptomyces-like strains isolated, 20 strains were found to be resistant to 50 mg/l of Cr(VI). In addition, two strains isolated from the estuarine sediment of Tokyo Bay were found to be resistant to a concentration of 150 mg/l of Cr(VI). Furthermore, one Cr(VI)-reducing strain was found to remove 60 mg/l of Cr(VI) within 1 week and was identified as Streptomyces thermocarboxydus based on 16S rRNA gene analysis. The comparative evaluation with the type strain S. thermocarboxydus NBRC 16323 showed that our isolated strain had higher ability to grow at 27 °C and reduce Cr(VI) at a NaCl concentration of 6.0 % at 27 °C compared with the type strain NBRC 16323. These results indicate that our isolated strain have a potential ability to remove Cr(VI) from contaminated, highly saline sources without heating.

  10. Screening of Marine Actinomycetes from Segara Anakan for Natural Pigment and Hydrolytic Activities

    Science.gov (United States)

    Asnani, A.; Ryandini, D.; Suwandri

    2016-02-01

    Marine actinomycetes have become sources of great interest to natural product chemistry due to their new chemical entities and bioactive metabolites. Since April 2010, we have screened actinobacteria from five sites that represent different ecosystems of Segara Anakan lagoon. In this present study we focus on specific isolates, K-2C which covers 1) actinomycetes identification based on morphology observation and 16S rRNA gene; 2) fermentation and isolation of pigment; 3) structure determination of pigment; and 4) hydrolytic enzymes characterization; Methodologies relevant to the studies were implemented accordingly. The results indicated that K-2C was likely Streptomyces fradiae strain RSU15, and the best fermentation medium should contain starch and casein with 21 days of incubation. The isolate has extracellular as well as intracellular pigments. Isolated pigments gave purple color with λmax of 529.00 nm. The pigment was structurally characterized. Interestingly, Streptomyces K-2C was able to produce potential hydrolytic enzymes such as amylase, cellulase, protease, lipase, urease, and nitrate reductase.

  11. Antimethicilin resistance agents from marine actinomycetes from soil sediments of Lagos Lagoon

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    Davies Olabisi Flora

    2015-03-01

    Full Text Available Objective: To evaluate the isolation of actinomycetes strains with potential for producing antimicrobials with high methicilin resistance capability. Methods: The soil samples were collected from four different locations of Lagos lagoon. The Actinomycetes were isolated from the samples by serial dilution using spread plate method. Isolates were selected based on their cultural characteristics as well as their Gram reaction and phenotypically and molecularly characterized Streptomyces sp. Isolates were inoculated in starch casein and Kuster’s broth media and secondary metabolites were screened for antimicrobial activity against the following microorganisms: methicillin resistant Staphylococcus aureus, Staphylococcus aureus ATCC 29213, Escherichia coli ATCC 29522, Pseudomonas aeruginosa ATCC 27853, Candida albicans, Enterococcus faecalis ATCC 29212. Coagulase-negative staphylococci isolated from HIV patients were also used (Staphylococcus warneri, Staphylococcus xylosus and Staphylococcus epidermidis. The antimicrobial metabolites of the isolates were identified using gas chromatography-mass spectrometer. Results: Extracts from isolates ULS12 and ULS13 showed antimicrobial activity against methicillin resistant Staphylococcus aureus while ULK3 inhibited Candida albicans only. The gas chromatography-mass spectrometer data analysis showed the antibiotic profile of these isolates. Conclusions: The isolates ULS12 and ULS13 were found to display the highest antimicrobial activity against the test organisms and could be a potential source of new antibiotics.

  12. Diversity and Antifungal Activity of Actinomycetes Symbiont Hard Coral Mucus of Genera Goniopora and Porites

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    Riyanti

    2016-12-01

    Full Text Available Screening new bioactive compounds from marine actinomycete organisms associated with corals (Goniopora and Porites can be an alternative method to discover the natural antifungal compounds. This study aims to determine the density and diversity of actinomycete symbionts based on repetitive sequence-based-polymerase chain reactions (rep-PCR and to discern the ability of antifungal activity of isolates symbiotic with hard coral mucus by using a pour plate method. A total of 143 isolates were obtained from the hard coral mucus of genera Goniopora and Porites. High genetic diversity was observed among the isolates. Ten isolates with different morphological characteristics were selected to extract its secondary metabolites and then followed by an antifungal test. The isolate with the code of SCAS324 was the one with the antifungal activity, marked by the formation of a very strong inhibition zone of 54.7±0.4 mm toward Aspergillus flavus and 49.2±2.7 mm toward Candida albicans. Antifungal screening showed that the antifungal activity of the isolate SCAS324 was three times as effective as the commercial antifungal.

  13. Targeted search for actinomycetes from nearshore and deep-sea marine sediments.

    Science.gov (United States)

    Prieto-Davó, Alejandra; Villarreal-Gómez, Luis J; Forschner-Dancause, Stephanie; Bull, Alan T; Stach, James E M; Smith, David C; Rowley, Dave C; Jensen, Paul R

    2013-06-01

    Sediment samples collected off the coast of San Diego were analyzed for actinomycete diversity using culture-independent techniques. Eight new operational taxonomic units (OTUs) in the Streptomycetaceae were identified as well as new diversity within previously cultured marine OTUs. Sequences belonging to the marine actinomycete genus Salinispora were also detected, despite the fact that this genus has only been reported from more tropical environments. Independent analyses of marine sediments from the Canary Basin (3814 m) and the South Pacific Gyre (5126 and 5699 m) also revealed Salinispora sequences providing further support for the occurrence of this genus in deep-sea sediments. Efforts to culture Salinispora spp. from these samples have yet to be successful. This is the first report of Salinispora spp. from marine sediments > 1100 m and suggests that the distribution of this genus is broader than previously believed. © 2013 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  14. Streptomyces temperate bacteriophage integration systems for stable genetic engineering of actinomycetes (and other organisms).

    Science.gov (United States)

    Baltz, Richard H

    2012-05-01

    ϕC31, ϕBT1, R4, and TG1 are temperate bacteriophages with broad host specificity for species of the genus Streptomyces. They form lysogens by integrating site-specifically into diverse attB sites located within individual structural genes that map to the conserved core region of streptomycete linear chromosomes. The target genes containing the ϕC31, ϕBT1, R4, and TG1 attB sites encode a pirin-like protein, an integral membrane protein, an acyl-CoA synthetase, and an aminotransferase, respectively. These genes are highly conserved within the genus Streptomyces, and somewhat conserved within other actinomycetes. In each case, integration is mediated by a large serine recombinase that catalyzes unidirectional recombination between the bacteriophage attP and chromosomal attB sites. The unidirectional nature of the integration mechanism has been exploited in genetic engineering to produce stable recombinants of streptomycetes, other actinomycetes, eucaryotes, and archaea. The ϕC31 attachment/integration (Att/Int) system has been the most widely used, and it has been coupled with the ϕBT1 Att/Int system to facilitate combinatorial biosynthesis of novel lipopeptide antibiotics in Streptomyces fradiae.

  15. In vitro actinomycete biofilm development and inhibition by the polyene antibiotic, nystatin, on IUD copper surfaces.

    Science.gov (United States)

    Shanmughapriya, Santhanam; Francis, Arumugam Lency; Kavitha, Senthil; Natarajaseenivasan, Kalimuthusamy

    2012-01-01

    The presence of intrauterine contraceptive devices (IUDs) gives a solid surface for attachment and an ideal niche for biofilm to form and flourish. Pelvic actinomycosis is often associated with the use of IUDs. Treatment of IUD-associated pelvic actinomycosis requires the immediate removal of the IUD. Therefore, this article presents in vitro evidence to support the use of novel antibiotics in the treatment of actinomycete biofilms. Twenty one clinical actinomycetes isolates from endocervical swabs of IUD wearers were assessed for their biofilm forming ability. An in vitro biofilm model with three isolates, Streptomyces strain A4, Nocardia strain C15 and Nocardia strain C17 was subjected to treatment with nystatin. Inhibition of biofilm formation by nystatin was found to be concentration dependent, with MBIC50 values in the range 0.08-0.16 mg ml(-1). Furthermore, at a concentration of 0.16 mg ml(-1), nystatin inhibited the twitching motility of the isolates, providing evidence for a possible mechanism of biofilm inhibition.

  16. Endophytic actinomycetes: a novel source of potential acyl homoserine lactone degrading enzymes.

    Science.gov (United States)

    Chankhamhaengdecha, Surang; Hongvijit, Suphatra; Srichaisupakit, Akkaraphol; Charnchai, Pattra; Panbangred, Watanalai

    2013-01-01

    Several Gram-negative pathogenic bacteria employ N-acyl-L-homoserine lactone (HSL) quorum sensing (QS) system to control their virulence traits. Degradation of acyl-HSL signal molecules by quorum quenching enzyme (QQE) results in a loss of pathogenicity in QS-dependent organisms. The QQE activity of actinomycetes in rhizospheric soil and inside plant tissue was explored in order to obtain novel strains with high HSL-degrading activity. Among 344 rhizospheric and 132 endophytic isolates, 127 (36.9%) and 68 (51.5%) of them, respectively, possessed the QQE activity. The highest HSL-degrading activity was at 151.30 ± 3.1 nmole/h/mL from an endophytic actinomycetes isolate, LPC029. The isolate was identified as Streptomyces based on 16S  rRNA gene sequence similarity. The QQE from LPC029 revealed HSL-acylase activity that was able to cleave an amide bond of acyl-side chain in HSL substrate as determined by HPLC. LPC029 HSL-acylase showed broad substrate specificity from C6- to C12-HSL in which C10HSL is the most favorable substrate for this enzyme. In an in vitro pathogenicity assay, the partially purified HSL-acylase efficiently suppressed soft rot of potato caused by Pectobacterium carotovorum ssp. carotovorum as demonstrated. To our knowledge, this is the first report of HSL-acylase activity derived from an endophytic Streptomyces.

  17. Isolation, screening and identification of novel isolates of Actinomycetes from India for antimicrobial applications

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    Vineeta Singh

    2016-12-01

    Full Text Available The search for novel bioactive compounds from the natural environment has been rapidly increased with the increase in multi-drug resistant (MDR pathogens. In the present study, the antimicrobial potential of novel actinomycetes has been evaluated by initial screening of six soil samples. Primary and secondary screening was performed against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Candida albicans, Candida tropicalis, Trichophyton rubrum, and other MDR bacterial and fungal test strains, and at the end thirteen active isolates were selected for further study. Microbial strains were identified on the basis of growth conditions and other biochemical characters. Five most active microbial strains were identified using 16S rRNA sequence homology and designated as Streptomyces xanthophaeus MTCC 11938, Streptomyces variabilis MTCC 12266, Streptomyces xanthochromogenes MTCC 11937, Streptomyces levis EU 124569 and Streptomyces sp. NCIM 5500. Four antibacterial and three antifungal compounds isolated from the above five isolates were purified and partially characterized using UV absorption and IR spectra. Two antibacterial metabolites, belong to chromone and peptide antibiotic, respectively. The antifungal compounds were found to be of non-polyene nature. In conclusion, we study the isolation of novel bacterial strains of actinomycetes for producing novel compounds having antibacterial and antifungal activities from the unexplored agro-ecological niches of India. Also, this study paves the way for further characterization of these isolates of Streptomyces sp. for their optimum utilization for antimicrobial purposes.

  18. Endophytic Actinomycetes: A Novel Source of Potential Acyl Homoserine Lactone Degrading Enzymes

    Directory of Open Access Journals (Sweden)

    Surang Chankhamhaengdecha

    2013-01-01

    Full Text Available Several Gram-negative pathogenic bacteria employ N-acyl-L-homoserine lactone (HSL quorum sensing (QS system to control their virulence traits. Degradation of acyl-HSL signal molecules by quorum quenching enzyme (QQE results in a loss of pathogenicity in QS-dependent organisms. The QQE activity of actinomycetes in rhizospheric soil and inside plant tissue was explored in order to obtain novel strains with high HSL-degrading activity. Among 344 rhizospheric and 132 endophytic isolates, 127 (36.9% and 68 (51.5% of them, respectively, possessed the QQE activity. The highest HSL-degrading activity was at 151.30±3.1 nmole/h/mL from an endophytic actinomycetes isolate, LPC029. The isolate was identified as Streptomyces based on 16S  rRNA gene sequence similarity. The QQE from LPC029 revealed HSL-acylase activity that was able to cleave an amide bond of acyl-side chain in HSL substrate as determined by HPLC. LPC029 HSL-acylase showed broad substrate specificity from C6- to C12-HSL in which C10HSL is the most favorable substrate for this enzyme. In an in vitro pathogenicity assay, the partially purified HSL-acylase efficiently suppressed soft rot of potato caused by Pectobacterium carotovorum ssp. carotovorum as demonstrated. To our knowledge, this is the first report of HSL-acylase activity derived from an endophytic Streptomyces.

  19. The genus Nonomuraea: A review of a rare actinomycete taxon for novel metabolites.

    Science.gov (United States)

    Sungthong, Rungroch; Nakaew, Nareeluk

    2015-05-01

    The genus Nonomuraea is a rare actinomycete taxon with a long taxonomic history, while its generic description was recently emended. The genus is less known among the rare actinomycete genera as its taxonomic position was revised several times. It can be found in diverse ecological niches, while most of its member species were isolated from soil samples. However, new trends to discover the genus in other habitats are increasing. Generic abundance of the genus was found to be dependent on geographical changes. Novel sources together with selective and invented isolation techniques might increase a chance to explore the genus and its novel candidates. Interestingly, some of its members have been revealed as a valuable source of novel metabolites for medical and industrial purposes. Broad-range of potent bioactive compounds including antimicrobial, anticancer, and antipsychotic substances, broad-spectrum antibiotics and biocatalysts can be synthesized by the genus. In order to investigate biosynthetic pathways of the bioactive compounds and self-resistant mechanisms to these compounds, the links from genes to metabolites have yet been needed for further discovery and biotechnological development of the genus Nonomuraea. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Marine sponge Craniella austrialiensis-associated bacterial diversity revelation based on 16S rDNA library and biologically active Actinomycetes screening, phylogenetic analysis.

    Science.gov (United States)

    Li, Z-Y; Liu, Y

    2006-10-01

    The aim of this study was to investigate the bacterial diversity associated with the sponge Craniella australiensis using a molecular strategy and isolating Actinomycetes with antimicrobial potentials. The bacterial diversity associated with South China Sea sponge C. austrialiensis was assessed using a 16S rDNA clone library alongside restriction fragment length polymorphism and phylogenetic analysis. It was found that the C. austrialiensis-associated bacterial community consisted of alpha, beta and gamma-Proteobacteria, Firmicutes, Bacteroidetes as well as Actinobacterium. Actinomycetes were isolated successfully using seawater medium with sponge extracts. According to the BLAST and phylogenetic analysis based on about 600-bp 16S rDNA sequences, 11 of the representative 23 isolates closely matched the Streptomyces sp. while the remaining 12 matched the Actinomycetales. Twenty Actinomycetes have antimicrobial potentials, of which 15 are found to possess broad-spectrum antimicrobial potentials. The sponge C. austrialiensis-associated bacterial community is very abundant including Proteobacteria, Firmicutes, Bacteroidetes and Actinobacterium while Actinomycetes is not predominant. Artificial seawater medium with sponge extracts is suitable for Actinomycetes isolation. Most of the isolated C. austrialiensis-associated Actinomycetes have a broad spectrum of antimicrobial activity. This study revealed the diversity of the bacterial community and the isolated Actinomycetes with antimicrobial potentials associated with sponge C. australiensis.

  1. Identification of the minimal replicon of plasmid pMEA300 of the methylotrophic actinomycete Amycolatopsis methanolica

    NARCIS (Netherlands)

    Vrijbloed, J.W.; Jelínková, M.; Hessels, G.I.; Dijkhuizen, L.

    1995-01-01

    The actinomycete Amycolatopsis methanolica contains a 13.3 kb plasmid (pMEA300), capable of enhancing the spontaneous mutation frequency of its host. Depending on the growth medium pMEA300 is not only maintained as an integrated element but can additionally be present as a multicopy, autonomously re

  2. Evolution of cyclizing 5-aminolevulinate synthases in the biosynthesis of actinomycete secondary metabolites: outcomes for genetic screening techniques.

    Science.gov (United States)

    Petříčková, Kateřina; Chroňáková, Alica; Zelenka, Tomáš; Chrudimský, Tomáš; Pospíšil, Stanislav; Petříček, Miroslav; Krištůfek, Václav

    2015-01-01

    A combined approach, comprising PCR screening and genome mining, was used to unravel the diversity and phylogeny of genes encoding 5-aminolevulinic acid synthases (ALASs, hemA gene products) in streptomycetes-related strains. In actinomycetes, these genes were believed to be directly connected with the production of secondary metabolites carrying the C5N unit, 2-amino-3-hydroxycyclopent-2-enone, with biological activities making them attractive for future use in medicine and agriculture. Unlike "classical" primary metabolism ALAS, the C5N unit-forming cyclizing ALAS (cALAS) catalyses intramolecular cyclization of nascent 5-aminolevulinate. Specific amino acid sequence changes can be traced by comparison of "classical" ALASs against cALASs. PCR screening revealed 226 hemA gene-carrying strains from 1,500 tested, with 87% putatively encoding cALAS. Phylogenetic analysis of the hemA homologs revealed strain clustering according to putative type of metabolic product, which could be used to select producers of specific C5N compound classes. Supporting information was acquired through analysis of actinomycete genomic sequence data available in GenBank and further genetic or metabolic characterization of selected strains. Comparison of 16S rRNA taxonomic identification and BOX-PCR profiles provided evidence for numerous horizontal gene transfers of biosynthetic genes or gene clusters within actinomycete populations and even from non-actinomycete organisms. Our results underline the importance of environmental and evolutionary data in the design of efficient techniques for identification of novel producers.

  3. Distribution and generic composition of culturable marine actinomycetes from the sediments of Indian continental slope of Bay of Bengal

    Science.gov (United States)

    Das, Surajit; Lyla, P. S.; Ajmal Khan, S.

    2008-05-01

    Actinomycetes population from continental slope sediment of the Bay of Bengal was studied. Samples were collected during two voyages of FORV Sagar Sampada in 2004 (May-June) and 2005 (July) respectively from 11 transects (each transect had ca. 200 m, 500 m, and 1 000 m depth stations). The physicochemical parameters of overlying water, and sediment samples were also recorded. The actinomycete population ranged from 5.17 to 51.94 CFU/g dry sediment weight and 9.38 to 45.22 CFU/g dry sediment weight during the two cruises respectively. No actinomycete colony was isolated from stations in 1 000 m depth. Two-way analysis of variance showed significant variation among stations (ANOVA two-way, P0.05). Three actinomycetes genera were identified. Streptomyces was found to be the dominating one in both the cruises, followed by Micromonospora, and Actinomyces. The spore of Streptomyces isolates showed the abundance in spiral spore chain. Spore surface was smooth. Multiple regression analysis revealed that the influencing physico-chemical factors were sediment pH, sediment temperature, TOC, porosity, salinity, and pressure. The media used in the present study was prepared with seawater. Thus, they may represent an autochthonous marine flora and deny the theory of land runoff carriage into the sea for adaptation to the salinity of the seawater and sediments.

  4. Different Physiological Roles of ATP- and PPi-Dependent Phosphofructokinase Isoenzymes in the Methylotrophic Actinomycete Amycolatopsis methanolica

    NARCIS (Netherlands)

    Alves, A.M.C.R.; Euverink, G.J.W.; Santos, H.; Dijkhuizen, L.

    2001-01-01

    Cells of the actinomycete Amycolatopsis methanolica grown on glucose possess only a single, exclusively PPi-dependent phosphofructokinase (PPi-PFK) (A. M. C. R. Alves, G. J. W. Euverink, H. J. Hektor, J. van der Vlag, W. Vrijbloed, D.H.A. Hondmann, J. Visser, and L. Dijkhuizen, J. Bacteriol. 176:682

  5. Use of the meganuclease I-SceI of Saccharomyces cerevisiae to select for gene deletions in actinomycetes.

    Science.gov (United States)

    Fernández-Martínez, Lorena T; Bibb, Mervyn J

    2014-11-18

    The search for new natural products is leading to the isolation of novel actinomycete species, many of which will ultimately require genetic analysis. Some of these isolates will likely exhibit low intrinsic frequencies of homologous recombination and fail to sporulate under laboratory conditions, exacerbating the construction of targeted gene deletions and replacements in genetically uncharacterised strains. To facilitate the genetic manipulation of such species, we have developed an efficient method to generate gene or gene cluster deletions in actinomycetes by homologous recombination that does not introduce any other changes to the targeted organism's genome. We have synthesised a codon optimised I-SceI gene for expression in actinomycetes that results in the production of the yeast I-SceI homing endonuclease which produces double strand breaks at a unique introduced 18 base pair recognition sequence. Only those genomes that undergo homologous recombination survive, providing a powerful selection for recombinants, approximately half of which possess the desired mutant genotype. To demonstrate the efficacy and efficiency of the system, we deleted part of the gene cluster for the red-pigmented undecylprodiginine complex of compounds in Streptomyces coelicolor M1141. We believe that the system we have developed will be broadly applicable across a wide range of actinomycetes.

  6. Azalomycin F4a 2-ethylpentyl ester, a new macrocyclic lactone, from mangrove actinomycete Streptomyces sp.211726

    Institute of Scientific and Technical Information of China (English)

    Gan Jun Yuan; Kui Hong; Hai Peng Lin; Jia Li

    2010-01-01

    Azalomycin F4a 2-ethylpentyl ester,a new 36-membered macrocyclic lactone antibiotic,was isolated from mangrove actinomycete Streptomyces sp.211726.Its structure was elucidated on the basis of spectroscopic data.The compound showed broad-spectrum antifungal activity and moderate cytotoxicity against human colon tumor cell HCT-116.

  7. A new naphthalenepropanoic acid analog from the marine-derived actinomycetes Micromonospora sp. HS-HM-036.

    Science.gov (United States)

    Gao, Mei-Yue; Qi, Huan; Li, Jian-Song; Zhang, Hui; Zhang, Ji; Wang, Ji-Dong; Xiang, Wen-Sheng

    2017-09-01

    A new naphthalenepropanoic acid analog (1) was isolated from the broth of the actinomycetes Micromonospora sp. HS-HM-036. The structure of compound 1 was determined based on MS and extensive NMR analysis. A preliminary investigation of the biological activity of compound 1 was also described.

  8. In vitro Antimicrobial Assay of Actinomycetes in Rice AgainstXanthomonas oryzae pv. oryzicola and as Potential Plant Growth Promoter

    Directory of Open Access Journals (Sweden)

    Erneeza Mohd Hata

    2015-12-01

    Full Text Available ABSTRACT The aim of this work was to invitro assay the antimicrobial activity of actinomycetes in rice against Xanthomonas oryzae pv. oryzicola and as potential plant growth promoter. A total of 92 actinomycete strains were isolated from different rice plant components and field locations. Of these, only 21.74% showed antagonistic activity against the Xoc pathogen. Molecular identification via 16s rRNA amplification revealed that 60% of the active antagonistic strains belonged to the genus Streptomyces. Isolates that demonstrated the highest antagonistic activity were also able to produce hydrolytic enzymes and plant growth-promoting hormones. Combination of preliminary screening based on in vitro antagonistic, hydrolytic enzyme and plant growth hormone activity facilitated the best selection of actinomycete candidates as evidenced by strains classification using cluster analysis (Ward's Method. Results from the preliminary screening showed that actinomycetes, especially Streptomycetes, could offer a promising source for both biocontrol and plant growth-promotion agents against BLS disease in rice.

  9. Retraction: Characterization of cellulolytic activities of newly isolated Thelephora sowerbyi from North-Western Himalayas on different lignocellulosic substrates.

    Science.gov (United States)

    Sharma, Deepika; Goel, Gunjan; Bansal, Saurabh; Mahajan, Rishi; Sharma, B M; Chauhan, Rajinder Singh

    2016-12-01

    Characterization of cellulolytic activities of newly isolated Thelephora sowerbyi from North-Western Himalayas on different lignocellulosic substrate J. Basic Microbiol. 2015, 55, 1-11 - DOI: 10.1002/jobm.201500107 The above article from the Journal of Basic Microbiology, published online on 08 June 2015 in Wiley Online Library as Early View (http://onlinelibrary.wiley.com/doi/10.1002/jobm.201500107/pdf), has been retracted by agreement between the authors, the Editor-in-Chief and Wiley-VCH GmbH & Co. KGaA. The retraction has been agreed because the microorganism studied in the described experiments has been identified as the fungus Cotylidia pannosa (Gene Accession No. KT008117) instead of Thelephora sowerbyi. The culture has been identified on the basis of the sequence of the amplified ITS region of the microorganism which was submitted by the authors to the NCBI database.

  10. Synergy between cellulolytic enzymes during the biodegradation of cellulose microfibrils measured using angle-scanning surface plasmon resonance (SPR) imaging

    Science.gov (United States)

    Raegen, Adam; Dion, Alexander; Reiter, Kyle; Clarke, Anthony; Lipkowski, Jacek; Dutcher, John

    2014-03-01

    The use of cellulosic ethanol, a promising emerging energy source, is limited by the energy intensive and costly step of first converting the cellulose fibers into their constituent glucose monomers. Industrial processes mimic those that occur in nature, using mixtures or ``cocktails'' of different classes of cellulolytic enzymes derived from fungi. Despite several decades of investigation, the molecular mechanisms for enzyme synergy remain poorly understood. To gain additional insight, we have used a custom angle-scanning surface plasmon resonance (SPR) imaging apparatus to obtain a sensitive measure of enzymatic degradation. By implementing a novel SPR data analysis procedure, we have been able to track the thickness and roughness of laterally heterogeneous cellulose microfibril-coated substrates as enzymatic degradation proceeds. This has allowed us to measure the synergistic actions of the different enzymes, providing data that are directly relevant to the cellulosic ethanol industry.

  11. Identification and molecular modeling of a family 5 endocellulase from Thermus caldophilus GK24, a cellulolytic strain of Thermus thermophilus

    Directory of Open Access Journals (Sweden)

    Dae-Sil Lee

    2006-12-01

    Full Text Available The genome of T. caldophilus GK24 was recently sequenced and annotated as 14contigs, equivalent to 2.3 mega basepairs (Mbp of DNA. In the current study, we identifieda unique 13.7 kbp DNA sequence, which included the endocellulase gene of T. caldophilusGK24, which did not appear to be present in the complete genomic sequence of the closelyrelated species T. thermophilus HB27 and HB8. Congo-red staining revealed a uniquephenotype of cellulose degradation by strain GK24 that was distinct from other closelyrelated Thermus strains. The results showed that strain GK24 is an aerobic, thermophilic,cellulolytic eubacterium which belongs to the group T. thermophilus. In order to understandthe mechanism of production of cellobiose in T. caldophilus GK24, a three-dimensionalmodel of the endocellulase, TcCel5A, was generated based on known crystal structures.Using this model, we carried out a flexible cellotetraose docking study.

  12. Structural insights into cellulolytic and chitinolytic enzymes revealing crucial residues of insect β-N-acetyl-D-hexosaminidase.

    Directory of Open Access Journals (Sweden)

    Tian Liu

    Full Text Available The chemical similarity of cellulose and chitin supports the idea that their corresponding hydrolytic enzymes would bind β-1,4-linked glucose residues in a similar manner. A structural and mutational analysis was performed for the plant cellulolytic enzyme BGlu1 from Oryza sativa and the insect chitinolytic enzyme OfHex1 from Ostrinia furnacalis. Although BGlu1 shows little amino-acid sequence or topological similarity with OfHex1, three residues (Trp(490, Glu(328, Val(327 in OfHex1, and Trp(358, Tyr(131 and Ile(179 in BGlu1 were identified as being conserved in the +1 sugar binding site. OfHex1 Glu(328 together with Trp(490 was confirmed to be necessary for substrate binding. The mutant E328A exhibited a 8-fold increment in K(m for (GlcNAc(2 and a 42-fold increment in K(i for TMG-chitotriomycin. A crystal structure of E328A in complex with TMG-chitotriomycin was resolved at 2.5 Å, revealing the obvious conformational changes of the catalytic residues (Glu(368 and Asp(367 and the absence of the hydrogen bond between E328A and the C3-OH of the +1 sugar. V327G exhibited the same activity as the wild-type, but acquired the ability to efficiently hydrolyse β-1,2-linked GlcNAc in contrast to the wild-type. Thus, Glu(328 and Val(327 were identified as important for substrate-binding and as glycosidic-bond determinants. A structure-based sequence alignment confirmed the spatial conservation of these three residues in most plant cellulolytic, insect and bacterial chitinolytic enzymes.

  13. Entomopathogenic marine actinomycetes as potential and low-cost biocontrol agents against bloodsucking arthropods.

    Science.gov (United States)

    Loganathan, Karthik; Kumar, Gaurav; Kirthi, Arivarasan Vishnu; Rao, Kokati Venkata Bhaskara; Rahuman, Abdul Abdul

    2013-11-01

    A novel approach to control strategies for integrated blood-feeding parasite management is in high demand, including the use of biological control agents. The present study aims to determine the efficacy of optimized crude extract of actinomycetes strain LK1 as biological control agent against the fourth-instar larvae of Anopheles stephensi and Culex tritaeniorhynchus (Diptera: Culicidae) and adults of Haemaphysalis bispinosa, Rhipicephalus (Boophilus) microplus (Acari: Ixodidae), and Hippobosca maculata (Diptera: Hippoboscidae). Antiparasitic activity was optimized using the Plackett-Burman method, and the design was developed using the software Design-Expert version 8.0.7.1. The production of the optimized crude actinomycetes LK1 strain extract was performed using response surface methodology to optimize the process parameters of protease inhibitor activity of marine actinobacteria for the independent variables like pH, temperature, glucose, casein, and NaCl at two levels (-1 and +1). The potential actinomycetes strain was identified as Saccharomonas spp., and the metamodeling surface simulation procedure was followed. It was studied using a computer-generated experimental design, automatic control of simulation experiments, and sequential optimization of the metamodels fitted to a simulation response surface function. The central composite design (CCD) used for the analysis of treatment showed that a second-order polynomial regression model was in good agreement with the experimental results at R (2) = 0.9829 (p < 0.05). The optimized values of the variables for antioxidant production were pH 6.00, glucose 1.3%, casein 0.09%, temperature 31.23 °C, and NaCl 0.10%. The LK1 strain-optimized crude extract was purified using reversed-phase high-pressure liquid chromatography, and the isolated protease inhibitor showed antiparasitic activity. The antiparasitic activity of optimized crude extract of LK1 was tested against larvae of A. stephensi (LC₅₀ = 31.82 ppm

  14. Isolasi Actinomycetes Laut Penghasil Metabolit Sekunder yang Aktif terhadap Sel Kanker A549

    Directory of Open Access Journals (Sweden)

    Rofiq Sunaryanto

    2010-12-01

    Full Text Available Telah dilakukan isolasi Actinomycetes laut yang mampu menghasilkan senyawa aktif citropeptin yang memiliki efek toksik terhadap sel kanker paru-paru A549. Isolasi dilakukan dengan menggunakan medium agar starch caseinyang ditambah dengan cycloheximidedan nistatin sebagai antifungi serta rifampisin dan nalidixic acids ebagai antibakteri. Sampel sedimen laut diperoleh dari pelabuhan Kamaishi-shi Iwate, Jepang pada kedalaman 5 meter. Dari 71 isolat yang diperoleh, hanya 9 isolat menunjukkan aktivitas terhadap sel kanker A549 pada konsentrasi 1 µg/200 µL. Hasil studi lebih lanjut menunjukkan bahwa isolat RS02-85 yang merupakan isolat terpilih adalah Streptomyces tsukubaensis dengan tingkat kemiripan 98%. Dari hasil identifikasi senyawa aktif, diduga senyawa tersebut adalah citropeptin dengan m/z (M+H+ 1035,4 g/mol dan rumus molekul C50H82N8O15

  15. Nocardia kroppenstedtii sp. nov., an actinomycete isolated from a lung transplant patient with a pulmonary infection.

    LENUS (Irish Health Repository)

    Jones, Amanda L

    2014-03-01

    A novel actinomycete, strain N1286(T), isolated from a lung transplant patient with a pulmonary infection, was provisionally assigned to the genus Nocardia. The strain had chemotaxonomic and morphological properties typical of members of the genus Nocardia and formed a distinct phyletic line in the Nocardia 16S rRNA gene tree. Isolate N1286(T) was most closely related to Nocardia farcinica DSM 43665(T) (99.8% gene sequence similarity) but could be distinguished from the latter by the low level of DNA-DNA relatedness. These strains were also distinguishable on the basis of a broad range of phenotypic properties. It is concluded that strain N1286(T) represents a novel species of the genus Nocardia for which the name Nocardia kroppenstedtii sp. nov. is proposed. The type strain is N1286(T) ( = DSM 45810(T) = NCTC 13617(T)).

  16. [Identification and analysis of an actinomycete strain suppressing Clavibacter michiganensis subsp, michiganensis].

    Science.gov (United States)

    Zhang, Yan; Zhang, Weihong; Wang, Songhong; Li, Yaning; Zhao, Zhiquan; Liu, Daqun; Yang, Wenxiang

    2009-07-01

    To identify and analyze bioactive compounds of an actinomycete strain Z-L-22 suppressing Clavibacter michiganensis subsp. michiganensis, the causal agent of bacterial canker of tomato. Morphological, biological and biochemical characterization, chemotaxonomy analysis and 16S rDNA sequences homology analysis were performed to identify the strain Z-L-22. Bioactive compounds were separated and retrieved by thin layer chromatography. Paper chromatography and confirmation tests were used to identify the antibiotic. PCR was carried out using the primers targeted to synthetase of the antibiotic. Strain Z-L-22 belonged to Streptomyces sp. and was similar to Streptomyces setonii. Two main bioactive components were isolated by thin layer chromatography, which were all identified as actinomycin. New actinomycin synthetase gene was cloned using the primers designed from actinomycin synthetase conserve domain. Strain Z-L-22 was classified as Streptomyces setonii. Actinomycin produced by Streptomyces setonii was first reported.

  17. Semi-solid-state fermentation: a promising alternative for neomycin production by the actinomycete Streptomyces fradiae.

    Science.gov (United States)

    Machado, Isabel; Teixeira, José A; Rodríguez-Couto, Susana

    2013-06-10

    The production of neomycin by the actinomycete Streptomyces fradiae, under semi-solid-state fermentation conditions was the main subject of this study. Two supports (nylon sponge and orange peelings) were tested in order to determine the most suitable one for the production of neomycin by the above-mentioned microorganism. Nylon sponge led to the highest neomycin production, reaching a maximum value of 13,903 μg/mL on the 10th day of cultivation. As a control, the same experiment was performed under submerged fermentation (SmF) conditions, without solid support. Here the production of neomycin by S. fradiae was about 55-fold lower (i.e. 250 μg/mL) than that obtained for SSF.

  18. Biodegradation of anthracene by a novel actinomycete, Microbacterium sp. isolated from tropical hydrocarbon-contaminated soil.

    Science.gov (United States)

    Salam, Lateef B; Obayori, Oluwafemi S; Olatoye, Nojeem O

    2014-01-01

    A novel anthracene-degrading Gram-positive actinomycete, Microbacterium sp. strain SL10 was isolated from a hydrocarbon-contaminated soil at a mechanical engineering workshop in Lagos, Nigeria. The polluted soil had an unusually high total hydrocarbon content of 157 g/kg and presence of various heavy metals. The isolate tolerated salt concentration of more than 4%. It resisted cefotaxime, streptomycin and ciprofloxacin, but susceptible to meropenem, linezolid and vancomycin. The isolate exhibited growth rate and doubling time of 0.82 days(-1) and 0.84 days, respectively on anthracene. It degraded 57.5 and 90.12% of anthracene within 12 and 21 days, respectively while the rate of anthracene utilization by the isolate was 4.79 mg l(-1) d(-1). To the best of our knowledge, this is the first report of isolation and characterization of anthracene-degrading Microbacterium sp.

  19. Metabolomics of the bio-degradation process of aflatoxin B1 by actinomycetes at an initial pH of 6.0

    National Research Council Canada - National Science Library

    Eshelli, Manal; Harvey, Linda; Edrada-Ebel, RuAngelie; McNeil, Brian

    2015-01-01

    .... Different processes have been used to degrade AFB1. In this study, biological degradation of AFB1 was carried out using three Actinomycete species, Rhodococcus erythropolis ATCC 4277, Streptomyces lividans TK 24...

  20. Himalomycin A and cycloheximide-producing marine actinomycete from Lagos Lagoon soil sediment

    Directory of Open Access Journals (Sweden)

    Davies Olabisi Flora

    2015-05-01

    Full Text Available Objective: To isolate and screen Actinomycetes from Lagos Lagoon soil sediments for antibiotic production. Methods: Soil samples were collected from four different locations of Lagos Lagoon and were dried for 2 weeks. Actinomycetes were isolated by serial dilution using spread plate method on starch casein and Kuster’s agar supplemented with 80 μg/mL cycloheximide to prevent fungal growth. The plates were incubated at 28 °C for 1-2 weeks. Isolates were selected based on their cultural characteristics as well as their Gram’s reaction and subcultured on same media for isolation and incubated at 28 °C for 3 days. Pure cultures were maintained on nutrient agar slants at 4 °C. Thereafter, they were inoculated into starch casein and Kuster’s broth media and incubated at 28 °C for 8 days. The resulting crude extracts were screened for antimicrobial activity against the following microorganisms: methicillin resistant Staphylococcus aureus, Staphylococcus aureus ATCC 29213, Escherichia coli ATCC 29522, Pseudomonas aeruginosa ATCC 27853, Candida albicans and Enterococcus faecalis ATCC 29212. Coagulasenegative staphylococci isolated from HIV patients were also used (Staphylococcus warneri, Staphylococcus xylosus and Staphylococcus epidermidis. Extraction of secondary metabolites was carried out and analysed using gas chromatography-mass spectrometer. Results: All the isolates displayed varying antimicrobial activity against at least one of the test organisms. Himalomycin A was identified in the extract from isolate ULS7. The gas chromatography-mass spectrometer data analysis showed the antibiotic profile of these isolates. Conclusions: The isolate ULS7 was found to display the highest antimicrobial activity against the test organisms.

  1. Himalomycin A and cycloheximide-producing marine actinomycete from Lagos Lagoon soil sediment

    Institute of Scientific and Technical Information of China (English)

    Davies Olabisi Flora; Adeleye IsaacAdeyemi; Wang Peng George

    2015-01-01

    Objective: To isolate and screen Actinomycetes from Lagos Lagoon soil sediments for antibiotic production. Methods: Soil samples were collected from four different locations of Lagos Lagoon and were dried for 2 weeks. Actinomycetes were isolated by serial dilution using spread plate method on starch casein and Kuster’s agar supplemented with 80 μg/mL cycloheximide to prevent fungal growth. The plates were incubated at 28 °C for 1-2 weeks. Isolates were selected based on their cultural characteristics as well as their Gram’s reaction and subcultured on same media for isolation and incubated at 28 °C for 3 days. Pure cultures were maintained on nutrient agar slants at 4 °C. Thereafter, they were inoculated into starch casein and Kuster’s broth media and incubated at 28 °C for 8 days. The resulting crude extracts were screened for antimicrobial activity against the following microorganisms: methicillin resistant Staphylococcus aureus, Staphylococcus aureus ATCC 29213, Escherichia coli ATCC 29522, Pseudomonas aeruginosa ATCC 27853, Candida albicans and Enterococcus faecalis ATCC 29212. Coagulase-negative staphylococci isolated from HIV patients were also used (Staphylococcus warneri, Staphylococcus xylosus and Staphylococcus epidermidis). Extraction of secondary metabolites was carried out and analysed using gas chromatography-mass spectrometer. Results: All the isolates displayed varying antimicrobial activity against at least one of the test organisms. Himalomycin A was identified in the extract from isolate ULS7. The gas chromatography-mass spectrometer data analysis showed the antibiotic profile of these isolates. Conclusions: The isolate ULS7 was found to display the highest antimicrobial activity against the test organisms.

  2. Marine actinomycetes: a new source of compounds against the human malaria parasite.

    Directory of Open Access Journals (Sweden)

    Jacques Prudhomme

    Full Text Available BACKGROUND: Malaria continues to be a devastating parasitic disease that causes the death of 2 million individuals annually. The increase in multi-drug resistance together with the absence of an efficient vaccine hastens the need for speedy and comprehensive antimalarial drug discovery and development. Throughout history, traditional herbal remedies or natural products have been a reliable source of antimalarial agents, e.g. quinine and artemisinin. Today, one emerging source of small molecule drug leads is the world's oceans. Included among the source of marine natural products are marine microorganisms such as the recently described actinomycete. Members of the genus Salinispora have yielded a wealth of new secondary metabolites including salinosporamide A, a molecule currently advancing through clinical trials as an anticancer agent. Because of the biological activity of metabolites being isolated from marine microorganisms, our group became interested in exploring the potential efficacy of these compounds against the malaria parasite. METHODS: We screened 80 bacterial crude extracts for their activity against malaria growth. We established that the pure compound, salinosporamide A, produced by the marine actinomycete, Salinispora tropica, shows strong inhibitory activity against the erythrocytic stages of the parasite cycle. Biochemical experiments support the likely inhibition of the parasite 20S proteasome. Crystal structure modeling of salinosporamide A and the parasite catalytic 20S subunit further confirm this hypothesis. Ultimately we showed that salinosporamide A protected mice against deadly malaria infection when administered at an extremely low dosage. CONCLUSION: These findings underline the potential of secondary metabolites, derived from marine microorganisms, to inhibit Plasmodium growth. More specifically, we highlight the effect of proteasome inhibitors such as salinosporamide A on in vitro and in vivo parasite development

  3. SCREENING, ISOLATION AND PURIFICATION OF ANTIBIOTIC(S FROM MARINE ACTINOMYCETES

    Directory of Open Access Journals (Sweden)

    Attimarad S L

    2012-06-01

    Full Text Available As marine environmental conditions are extremely different from terrestrial ones, it is surmised that marine actinomycetes might produce novel bioactive compounds. Hence marine sediments, collected from the coastal areas of Gokharna and Muradeshwara of Karnataka state, were screened. Seventeen isolates were obtained on starch-casein agar media by soil dilution technique. However, only six isolates namely SUN-A2, SUN-A3, SUN-A4, SUN-A5, SUN-A7 and SUN-A15 showed significant antibacterial activity against both gram-positive and gram-negative bacteria. Further studies were carried out with the most active SUN-A2. Optimization of media, temperature and pH by shake flask fermentation indicated starch-casein, 28o C and pH 7 to be suitable for SUN-A2. The production of antibiotics began after 24 h reached maximum at 72 h and maintained at the same level up to 120 h. Ethyl acetate was used to extract antibacterial compounds from the culture filtrate. TLC was done on silica gel using ethyl acetate: methanol (6:4 and direct bioautography showed the presence of two active substance, one with Rf 0.8 more active than the other with Rf 0.4. Further purification is done by column chromatography using a mixture of dicholoromethane and ethyl acetate. The findings from this investigation reveal that the strain SUN-A2 in order exhibited superior antimicrobial activity to other sediment isolates of actinomycetes.

  4. Antibacterial activity of Pseudonocardia sp. JB05, a rare salty soil actinomycete against Staphylococcus aureus.

    Science.gov (United States)

    Jafari, Nesa; Behroozi, Reza; Farajzadeh, Davoud; Farsi, Mohammad; Akbari-Noghabi, Kambiz

    2014-01-01

    Staphylococcus aureus is a Gram-positive bacterium that causes many harmful and life-threatening diseases. Some strains of this bacterium are resistant to available antibiotics. This study was designed to evaluate the ability of indigenous actinomycetes to produce antibacterial compounds against S. aureus and characterize the structure of the resultant antibacterial compounds. Therefore, a slightly modified agar well diffusion method was used to determine the antibacterial activity of actinomycete isolates against the test microorganisms. The bacterial extracts with antibacterial activity were fractionated by silica gel and G-25 sephadex column chromatography. Also, the active fractions were analyzed by thin layer chromatography. Finally, the partial structure of the resultant antibacterial compound was characterized by Fourier transform infrared spectroscopy. One of the isolates, which had a broad spectrum and high antibacterial activity, was designated as Pseudonocardia sp. JB05, based on the results of biochemical and 16S rDNA gene sequence analysis. Minimum inhibitory concentration for this bacterium was 40 AU mL(-1) against S. aureus. The antibacterial activity of this bacterium was stable after autoclaving, 10% SDS, boiling, and proteinase K. Thin layer chromatography, using anthrone reagent, showed the presence of carbohydrates in the purified antibacterial compound. Finally, FT-IR spectrum of the active compound illustrated hydroxyl groups, hydrocarbon skeleton, and double bond of polygenic compounds in its structure. To the best of our knowledge, this is the first report describing the efficient antibacterial activity by a local strain of Pseudonocardia. The results presented in this work, although at the initial stage in bioactive product characterization, will possibly contribute toward the Pseudonocardia scale-up for the production and identification of the antibacterial compounds.

  5. Antibiotic Producing Potentials of Three Freshwater Actinomycetes Isolated from the Eastern Cape Province of South Africa

    Directory of Open Access Journals (Sweden)

    Timothy Sibanda

    2010-07-01

    Full Text Available Crude extracts of three actinomycetes species belonging to Saccharopolyspora (TR 046 and TR 039 and Actinosynnema (TR 024 genera were screened for antibacterial activities against a panel of several bacterial strains. The extracts showed antibacterial activities against both gram-negative and gram-positive test bacteria with inhibition zones ranging from 8 to 28 mm (TR 046; 8 to15 mm (TR 039; and 10 to 13 mm (TR 024. The minimum inhibitory concentrations ranged from 0.078 to 10 mg/mL (TR 046; 5 to >10 mg/mL (TR 039; and 1.25 to 5 mg/mL (TR 024. Time-kill studies revealed that crude extract of TR 046 showed strong bactericidal activity against Bacillus pumilus (ATCC14884, reducing the bacterial load by 104 cfu/mL and 102 cfu/mL at 4× MIC and 2× MIC, respectively, after 6 h of exposure. Similarly, against Proteus vulgaris (CSIR 0030, crude extract of TR 046 achieved a 0.9log10 and 0.13log10 cfu/mL reduction at 5 mg/mL (4× MIC and 1.25 mg/mL (2× MIC after 12 h of exposure. The extract was however weakly bactericidal against two environmental bacterial strains (Klebsiella pneumoniae and Staphylococcus epidermidis; and against Pseudomonas aeruginosa (ATCC 19582: the extract showed bacteriostatic activities at all concentrations tested. These freshwater actinomycetes appear to have immense potential as a source of new antibacterial compound(s.

  6. Effect of Feeding Palm Oil By-Products Based Diets on Total Bacteria, Cellulolytic Bacteria and Methanogenic Archaea in the Rumen of Goats

    OpenAIRE

    Abdelrahim Abubakr; Abdul Razak Alimon; Halimatun Yaakub; Norhani Abdullah; Michael Ivan

    2014-01-01

    Rumen microorganisms are responsible for digestion and utilization of dietary feeds by host ruminants. Unconventional feed resources could be used as alternatives in tropical areas where feed resources are insufficient in terms of quality and quantity. The objective of the present experiment was to evaluate the effect of diets based on palm oil (PO), decanter cake (DC) or palm kernel cake (PKC) on rumen total bacteria, selected cellulolytic bacteria, and methanogenic archaea. Four diets: cont...

  7. Study of the diversity of culturable actinomycetes in the North Pacific and Caribbean coasts of Costa Rica

    Science.gov (United States)

    Solano, Godofredo; Rojas-Jiménez, Keilor; Jaspars, Marcel

    2011-01-01

    In this study, 137 actinomycetes were isolated from subtidal marine sediments in the North Pacific and Caribbean coasts of Costa Rica. Bioinformatics analysis of the 16S rRNA gene sequences assigned the isolates to 15 families and 21 genera. Streptomyces was the dominant genus while the remaining 20 genera were poorly represented. Nearly 70% of the phylotypes presented a coastal-restricted distribution whereas the other 30% were common inhabitants of both shores. The coastal tropical waters of Costa Rica showed a high diversity of actinomycetes, both in terms of the number of species and phylogenetic composition, although significant differences were observed between and within shores. The observed pattern of species distribution might be the result of several factors including the characteristics of the ecosystems, presence of endemic species and the influence of terrestrial runoff. PMID:19365710

  8. Clear felling and burning effects on soil nitrogen transforming bacteria and actinomycetes population in Chittagong University campus, Bangladesh

    Institute of Scientific and Technical Information of China (English)

    S.M.Sirajul Haque; Rahima Ferdoshi; Sohag Miah; M.Nural Anwar

    2012-01-01

    The effect of forests clear felling and associated burning on the population of soil nitrogen transforming bacteria and actinomycetes are reported at three pair sites of Chittagong University campus,Bangladesh in monsoon tropical climate.Clear felled area or burnt site and 15-21 year mixed plantation of native and exotic species,situated side by side on low hill having Typic Dystrochrepts soil was represented at each pair site.At all the three pair sites,clear felled area or burnt site showed very significantly (p≤0.001) lower population of actinomycetes,Rhizobium,Nitrosomonas,Nitrobacter and ammonifying as well as denitrifying bacteria compared to their adjacent mixed plantation.From environmental consideration,this finding has implication in managing natural ecosystem.

  9. Evolution of cyclizing 5-aminolevulinate synthases in the biosynthesis of actinomycete secondary metabolites: Outcomes for genetic screening techniques

    Directory of Open Access Journals (Sweden)

    Katerina ePetrickova

    2015-08-01

    Full Text Available A combined approach, comprising PCR screening and genome mining, was used to unravel the diversity and phylogeny of genes encoding 5-aminolevulinic acid synthases (ALASs, hemA gene products in streptomycetes-related strains. In actinomycetes, these genes were believed to be directly connected with the production of secondary metabolites carrying the C5N unit, 2-amino-3-hydroxycyclopent-2-enone, with biological activities making them attractive for future use in medicine and agriculture. Unlike classical primary metabolism ALAS, the C5N unit-forming cALAS (cyclizing ALAS catalyses intramolecular cyclization of nascent 5-aminolevulinate. Specific amino acid sequence changes can be traced by comparison of classical ALASs against cALASs. PCR screening revealed 226 hemA gene-carrying strains from 1,500 tested, with 87 % putatively encoding cALAS. Phylogenetic analysis of the hemA homologues revealed strain clustering according to putative type of metabolic product, which could be used to select producers of specific C5N compound classes. Supporting information was acquired through analysis of actinomycete genomic sequence data available in GeneBank and further genetic or metabolic characterization of selected strains. Comparison of 16S rRNA taxonomic identification and BOX-PCR profiles provided evidence for numerous horizontal gene transfers of biosynthetic genes or gene clusters within actinomycete populations and even from non-actinomycete organisms. Our results underline the importance of environmental and evolutionary data in the design of efficient techniques for identification of novel producers.

  10. Distribution and generic composition of culturable marine actinomycetes from the sediments of Indian continental slope of Bay of Bengal

    Institute of Scientific and Technical Information of China (English)

    Surajit DAS; P.S.LYLA; S.AJMAL KHAN

    2008-01-01

    Actinomycetes population from continental slope sediment of the Bay of Bengal was studied.Samples were collected during two voyages of FORV Sagar Sampada in 2004 (May-June) and 2005 (July) respectively from 11 transects (each transect had ca.200m,500m,and 1000m depth stations).The physicochemical parameters of overlying water,and sediment samples were also recorded.The actinomycete population ranged from 5.17 to 51.94 CFU/g dry sediment weight and 9.38 to 45.22 CFU/g dry sediment weight during the two cruises respectively.No actinomycete colony was isolated from stations in 1000m depth.Two-way analysis of variance showed significant variation among stations (ANOVA two-way,P0.05),but no significance was found between the two cruises (ANOVA two-way,P0.05).Three actinomycetes genera were identified.Streptomyces was found to be the dominating one in both the cruises,followed by Micromonospora,and Actinomyces.The spore of Streptomyces isolates showed the abundance in spiral spore chain.Spore surface was smooth.Multiple regression analysis revealed that the influencing physico-chemical factors were sediment pH,sediment temperature,TOC,porosity,salinity,and pressure.The media used in the present study was prepared with seawater.Thus,they may represent an autochthonous marine flora and deny the theory of land runoff carriage into the sea for adaptation to the salinity of the seawater and sediments.

  11. Transformation of 2,4,6-trinitrotoluene (TNT) by actinomycetes isolated from TNT-contaminated and uncontaminated environments

    Energy Technology Data Exchange (ETDEWEB)

    Pasti-Grigsby, M.B.; Lewis, T.A.; Crawford, D.L.; Crawford, R.L. [Univ. of Idaho, Moscow, ID (United States)

    1996-03-01

    Biotransformation of TNT has been reported under both aerobic and anaerobic conditions. Actinomycetes are important decomposers in composts. This study examines the tolerance of acitomycete cultures, isolated from both TNT-contaminated and uncontaminated environments for different concentrations to TNT, determined how selected isolates transform TNT, and examined whether such TNT transformations were constitutive or induced by exposure to TNT. 33 refs., 1 figs., 1 tab.

  12. Evaluation of antimicrobial activity of the endophytic actinomycete R18(6 against multiresistant Gram-negative bacteria

    Directory of Open Access Journals (Sweden)

    Tiele Carvalho

    2016-03-01

    Full Text Available Endophytic actinomycetes are promising sources of antimicrobial substances. This study evaluates the activity of metabolites produced by the endophytic actinomycete R18(6 against Gram-negative bacteria multiresistant to antimicrobials. R18(6 isolate was grown in submerged cultures under different conditions: carbon source, temperature, pH and incubation time to optimize antimicrobials production. The actinomycete grown in base medium supplemented with 1% glucose, pH 6.5 and incubation at 30 ºC for 96 h with shaking at 100 rpm, exhibited the highest activity against the used Gram-negative bacteria. Minimum inhibitory concentration (MIC of the crude extract produced by the microorganism varied between 1/32 and 1/256. It had bactericide or bacteriostatic activity, depending on the Gram-negative organism. The active extract was stable at high temperatures, and unstable in medium containing proteolytic enzymes. Micromorphology of R18(6 was investigated by optical and scan microscopy, revealing that it was morphologically similar to the genusStreptomyces.

  13. Degradative crystal–chemical transformations of clay minerals under the influence of cyanobacterium-actinomycetal symbiotic associations

    Directory of Open Access Journals (Sweden)

    Ekaterina Ivanova

    2014-04-01

    Full Text Available Cyanobacteria and actinomycetes are essential components of soil microbial community and play an active role in ash elements leaching from minerals of the parent rock. Content and composition of clay minerals in soil determine the sorption properties of the soil horizons, water-holding capacity of the soil, stickiness, plasticity, etc. The transformative effect of cyanobacterial–actinomycetes associations on the structure of clay minerals – kaolinite, vermiculite, montmorillonite, biotite and muscovite – was observed, with the greatest structural lattice transformation revealed under the influence of association in comparison with monocultures of cyanobacterium and actinomycete. The range of the transformative effect depended both on the type of biota (component composition of association and on the crystal–chemical parameters of the mineral itself (trioctahedral mica – biotite, was more prone to microbial degradation than the dioctahedral – muscovite. The formation of the swelling phase – the product of biotite transformation into the mica–vermicullite mixed-layered formation was revealed as a result of association cultivation. Crystal chemical transformation of vermiculite was accompanied by the removal of potassium (К, magnesium (Mg and aluminum (Al from the crystal lattice. The study of such prokaryotic communities existed even in the early stages of the Earth's history helps to understand the causes and nature of the transformations undergone by the atmosphere, hydrosphere and lithosphere of the planet.contribution of treatments on structure induces and model parameters are discussed in the paper.

  14. Cellulolytic potential of a novel strain of Paenibacillus sp. isolated from the armored catfish Parotocinclus maculicauda gut

    Directory of Open Access Journals (Sweden)

    André L. M. de Castro

    2011-12-01

    Full Text Available A cellulolytic bacterial strain, designated P118, isolated from the gut of the tropical fish Parotocinclus maculicauda was identified as belonging to the genus Paenibacillus based on phenotypic and chemotaxonomic characteristics and the 16S rRNA gene sequence. The novel strain was Gram-positive, spore-forming and rod-shaped. Catalase but not oxidase was produced. Carboxymethylcellulose was hydrolyzed but starch or gelatin was not. Acetoin production was negative whereas nitrate reduction and urease production were positive. Many carbohydrates served as carbon sources for growth. MK-7 was the predominant isoprenoid quinone. Anteiso-C15:0 (38.73% and C16:0 (20.85% were the dominant cellular fatty acids. Strain P118 was closely related to Paenibacillus amylolyticus NRRL NRS-290, P. pabuli HSCC 492, P. tundrae Ab10b, P. xylanexedens B22a, and P. tylopili MK2 with 98.3-98.8% 16S rRNA gene sequence similarity. The results presented here suggest that strain P118 represents a novel species of the genus Paenibacillus and it is a potential strain for further studies concerning its role in the production of industrially important products from cellulosic biomass.

  15. Caldicellulosiruptor obsidiansis sp. nov., an anaerobic, extremely thermophilic, cellulolytic bacterium isolated from Obsidian Pool, Yellowstone National Park

    Energy Technology Data Exchange (ETDEWEB)

    Hamilton-Brehm, Scott [ORNL; Elkins, James G [ORNL; Phelps, Tommy Joe [ORNL; Keller, Martin [ORNL; Carroll, Sue L [ORNL; Allman, Steve L [ORNL; Podar, Mircea [ORNL; Mosher, Jennifer J [ORNL; Vishnivetskaya, Tatiana A [ORNL

    2010-01-01

    A novel, obligately anaerobic, extremely thermophilic, cellulolytic bacterium, designated OB47T, was isolated from Obsidian Pool, Yellowstone National Park, WY, USA. The isolate was a non-motile, non-spore forming, Gram-positive rod approximately 2 m long by 0.2 m wide and grew at temperatures between 55-85oC with the optimum at 78oC. The pH range for growth was 6.0-8.0 with values of near 7.0 being optimal. Growth on cellobiose produced the fastest specific growth rates at 0.75 hr-1. The organism also displayed fermentative growth on glucose, maltose, arabinose, fructose, starch, lactose, mannose, sucrose, galactose, xylose, arabinogalactan, Avicel, xylan, filter paper, processed cardboard, pectin, dilute acid-pretreated switchgrass and Populus. OB47T was unable to grow on mannitol, fucose, lignin, Gelrite, acetate, glycerol, ribose, sorbital, carboxymethylcellulose and casein. Yeast extract stimulated growth and thiosulfate, sulfate, nitrate, and sulfur were not reduced. Fermentation end products were mainly acetate, H2, and CO2 although lactate and ethanol were produced in 5 l batch fermentations. The G+C content of the DNA was 35 mol% and sequence analysis of the small subunit ribosomal RNA gene placed OB47T within the genus Caldicellulosiruptor. Based on its phylogenetic and phenotypic properties, the isolate is proposed to be designated Caldicellulosiruptor obsidiansis sp. nov. and OB47T is the type stain (ATCC = ____, JCM = ____).

  16. PRODUCTION AND CHARACTERIZATION OF CELLULOLYTIC ENZYMES BY ASPERGILLUS NIGER AND RHIZOPUS SP . BY SOLID STATE FERMENTATION OF PRICKLY PEAR

    Directory of Open Access Journals (Sweden)

    TAMIRES CARVALHO DOS SANTOS

    2016-01-01

    Full Text Available Prickly palm cactus husk was used as a solid - state fermentation support substrate for the production of cellulolytic enzymes using Aspergillus niger and Rhizopus sp. A Box - Behnken design was used to evaluate the effects of water activity, fermentation time and temperature on endoglucanase and total cellulase production. Response Surface Methodology showed that optimum conditions for endoglucanase production were achieved at after 70.35 h of fermentation at 29.56°C and a water activity of 0.875 for Aspergillus niger and after 68.12 h at 30.41°C for Rhizopus sp. Optimum conditions for total cellulase production were achieved after 74.27 h of fermentation at 31.22°C for Aspergillus niger and after 72.48 h and 27.86°C for Rhizopus sp . Water activity had a significant effect on Aspergillus niger endoglucanase production only. In industrial applications, enzymatic characterization is important for optimizing variables such as temperature and pH. In this study we showed that endoglucanase and total cellulase had a high level of thermostability and pH stability in all the enzymatic extracts. Enzymatic deactivation kinetic experiments indicated that the enzymes remained active after the freezing of the crude extract. Based on the results, bioconversion of cactus is an excellent alternative for the production of thermostable enzymes.

  17. Cellulolytic enzymes production by utilizing agricultural wastes under solid state fermentation and its application for biohydrogen production.

    Science.gov (United States)

    Saratale, Ganesh D; Kshirsagar, Siddheshwar D; Sampange, Vilas T; Saratale, Rijuta G; Oh, Sang-Eun; Govindwar, Sanjay P; Oh, Min-Kyu

    2014-12-01

    Phanerochaete chrysosporium was evaluated for cellulase and hemicellulase production using various agricultural wastes under solid state fermentation. Optimization of various environmental factors, type of substrate, and medium composition was systematically investigated to maximize the production of enzyme complex. Using grass powder as a carbon substrate, maximum activities of endoglucanase (188.66 U/gds), exoglucanase (24.22 U/gds), cellobiase (244.60 U/gds), filter paperase (FPU) (30.22 U/gds), glucoamylase (505.0 U/gds), and xylanase (427.0 U/gds) were produced under optimized conditions. The produced crude enzyme complex was employed for hydrolysis of untreated and mild acid pretreated rice husk. The maximum amount of reducing sugar released from enzyme treated rice husk was 485 mg/g of the substrate. Finally, the hydrolysates of rice husk were used for hydrogen production by Clostridium beijerinckii. The maximum cumulative H2 production and H2 yield were 237.97 mL and 2.93 mmoL H2/g of reducing sugar, (or 2.63 mmoL H2/g of cellulose), respectively. Biohydrogen production performance obtained from this work is better than most of the reported results from relevant studies. The present study revealed the cost-effective process combining cellulolytic enzymes production under solid state fermentation (SSF) and the conversion of agro-industrial residues into renewable energy resources.

  18. Assessment of multi-enzyme operon engineering of tobacco chloroplast genome for high-level simultaneous expression of cellulolytic enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Kolotilin, I. [Agriculture and Agri-Food Canada, London, ON (Canada); Pereira, E.O.; Menassa, R. [Western Ontario Univ., London, ON (Canada). Dept. of Biology; Agriculture and Agri-Food Canada, London, ON (Canada)

    2009-07-01

    The use of biofuels as an environmentally-sound substitute for depleting fossil fuels was discussed. Commercially produced biofuels are generated primarily from starch or sugar and supply only a small fraction of global fuel requirements. Although cellulosic biomass can serve as an abundant and renewable source of fermentable sugars, the cost of converting biomass to fuel is too high. Plant genetic engineering techniques are more economical for producing recombinant proteins because of the low-cost of the growing bioreactors. The transformation of the tobacco chloroplast genome has proven to be very prolific in terms of recombinant protein yield, which typically reaches 10 to 20 per cent of total soluble protein. In addition, plastid transcription-translation machinery allows for the simultaneous expression of several genes from artificial operons, providing the potential to engineer several proteins in one transformation step. The purpose of this study was to produce transplastomic tobacco plants bearing single genes as well as operons of cell wall-degrading enzymes for high-level expression. An attempt was made to reproduce an engineering approach in tobacco chloroplasts to generate a potent mini-cellulosome. The resulting enzymes were evaluated for their ability to degrade biomass. The study also examined the feasibility of using crude extracts of highly-expressing plants as an additive in the biomass fermentation process. The productivity of transplastomic plants was compared with plants transiently expressing cellulolytic enzymes directed to other cellular compartments.

  19. Caldicellulosiruptor obsidiansis sp. nov., an anaerobic, extremely thermophilic, cellulolytic bacterium isolated from Obsidian Pool, Yellowstone National Park.

    Science.gov (United States)

    Hamilton-Brehm, Scott D; Mosher, Jennifer J; Vishnivetskaya, Tatiana; Podar, Mircea; Carroll, Sue; Allman, Steve; Phelps, Tommy J; Keller, Martin; Elkins, James G

    2010-02-01

    A novel, obligately anaerobic, extremely thermophilic, cellulolytic bacterium, designated OB47(T), was isolated from Obsidian Pool, Yellowstone National Park, WY. The isolate was a nonmotile, non-spore-forming, Gram-positive rod approximately 2 microm long by 0.2 microm wide and grew at temperatures between 55 and 85 degrees C, with the optimum at 78 degrees C. The pH range for growth was 6.0 to 8.0, with values of near 7.0 being optimal. Growth on cellobiose produced the fastest specific growth rate at 0.75 h(-1). The organism also displayed fermentative growth on glucose, maltose, arabinose, fructose, starch, lactose, mannose, sucrose, galactose, xylose, arabinogalactan, Avicel, xylan, filter paper, processed cardboard, pectin, dilute acid-pretreated switchgrass, and Populus. OB47(T) was unable to grow on mannitol, fucose, lignin, Gelrite, acetate, glycerol, ribose, sorbitol, carboxymethylcellulose, and casein. Yeast extract stimulated growth, and thiosulfate, sulfate, nitrate, and sulfur were not reduced. Fermentation end products were mainly acetate, H2, and CO2, although lactate and ethanol were produced in 5-liter batch fermentations. The G+C content of the DNA was 35 mol%, and sequence analysis of the small subunit rRNA gene placed OB47(T) within the genus Caldicellulosiruptor. Based on its phylogenetic and phenotypic properties, the isolate is proposed to be designated Caldicellulosiruptor obsidiansis sp. nov. and OB47 is the type strain (ATCC BAA-2073).

  20. Caldicellulosiruptor obsidiansis sp. nov., an Anaerobic, Extremely Thermophilic, Cellulolytic Bacterium Isolated from Obsidian Pool, Yellowstone National Park▿

    Science.gov (United States)

    Hamilton-Brehm, Scott D.; Mosher, Jennifer J.; Vishnivetskaya, Tatiana; Podar, Mircea; Carroll, Sue; Allman, Steve; Phelps, Tommy J.; Keller, Martin; Elkins, James G.

    2010-01-01

    A novel, obligately anaerobic, extremely thermophilic, cellulolytic bacterium, designated OB47T, was isolated from Obsidian Pool, Yellowstone National Park, WY. The isolate was a nonmotile, non-spore-forming, Gram-positive rod approximately 2 μm long by 0.2 μm wide and grew at temperatures between 55 and 85°C, with the optimum at 78°C. The pH range for growth was 6.0 to 8.0, with values of near 7.0 being optimal. Growth on cellobiose produced the fastest specific growth rate at 0.75 h−1. The organism also displayed fermentative growth on glucose, maltose, arabinose, fructose, starch, lactose, mannose, sucrose, galactose, xylose, arabinogalactan, Avicel, xylan, filter paper, processed cardboard, pectin, dilute acid-pretreated switchgrass, and Populus. OB47T was unable to grow on mannitol, fucose, lignin, Gelrite, acetate, glycerol, ribose, sorbitol, carboxymethylcellulose, and casein. Yeast extract stimulated growth, and thiosulfate, sulfate, nitrate, and sulfur were not reduced. Fermentation end products were mainly acetate, H2, and CO2, although lactate and ethanol were produced in 5-liter batch fermentations. The G+C content of the DNA was 35 mol%, and sequence analysis of the small subunit rRNA gene placed OB47T within the genus Caldicellulosiruptor. Based on its phylogenetic and phenotypic properties, the isolate is proposed to be designated Caldicellulosiruptor obsidiansis sp. nov. and OB47 is the type strain (ATCC BAA-2073). PMID:20023107

  1. Characterization of cellulolytic enzyme system of Schizophyllum commune mutant and evaluation of its efficiency on biomass hydrolysis.

    Science.gov (United States)

    Sornlake, Warasirin; Rattanaphanjak, Phatcharamon; Champreda, Verawat; Eurwilaichitr, Lily; Kittisenachai, Suthathip; Roytrakul, Sittiruk; Fujii, Tatsuya; Inoue, Hiroyuki

    2017-07-01

    Schizophyllum commune is a basidiomycete equipped with an efficient cellulolytic enzyme system capable of growth on decaying woods. In this study, production of lignocellulose-degrading enzymes from S. commune mutant G-135 (SC-Cel) on various cellulosic substrates was examined. The highest cellulase activities including CMCase, FPase, and β-glucosidase were obtained on Avicel-PH101 while a wider range of enzymes attacking non-cellulosic polysaccharides and lignin were found when grown on alkaline-pretreated biomass. Proteomic analysis of SC-Cel also revealed a complex enzyme system comprising seven glycosyl hydrolase families with an accessory carbohydrate esterase, polysaccharide lyase, and auxiliary redox enzymes. SC-Cel obtained on Avicel-PH101 effectively hydrolyzed all agricultural residues with the maximum glucan conversion of 98.0% using corn cobs with an enzyme dosage of 5 FPU/g-biomass. The work showed potential of SC-Cel on hydrolysis of various herbaceous biomass with enhanced efficiency by addition external β-xylosidase.

  2. Comparative efficacy of macrolides containing marine actinomycetes formulation versus ciprofloxacin ophthalmic solution in controlling Pseudomonas aeruginosa induced conjunctivitis on rabbit model

    Directory of Open Access Journals (Sweden)

    Femina Wahaab

    2015-06-01

    Full Text Available The main objective of this study was to evaluate the antimicrobial activity and anti-inflammatory activity of marine actinomycetes extract against ocular pathogen Pseudomonas aeruginosa. Actinomycetes isolated from Rameswaram coastal region, Tamilnadu, India were initially screened by primary screening and secondary screening against ocular pathogen P. aeruginosa. Followed by anti-conjunctivitis efficacy of actinomycetes ethyl acetate extract formulation versus ciprofloxacin ophthalmic solution was evaluated using rabbit as animal model. The bioactive compounds present in the best actinomycetes extract was identified by HPTLC and GC–MS analysis. Finally the screened best actinomycetes was identified by 16S rRNA sequencing method. In primary screening 28 actinomycetes that inhibited the growth of P. aeruginosa were taken for secondary screening. In secondary screening RAM24C2 extract had maximum activity against P. aeruginosa. In vivo study of conjunctivitis developed rabbits treated with RAM24C2 extract formulation showed the best clinical cure than ciprofloxacin ophthalmic solution. The RAM24C2 extract was chromatographically characterized and found to contain macrolides. In addition, the effective major pivotal molecule in the extract was detected as 1, 2 benzene dicarboxylic acid and Bis (2-ethylhexyl phthalate by GC–MS analysis. The RAM24C2 strain was identified as Streptomyces sp. MAD01 and the sequence was submitted in NCBI with accession number JX050218. From our study it is found that the ethyl acetate extract obtained from marine actinomycetes is effective against ocular pathogen P. aeruginosa. Compared to ciprofloxacin ophthalmic solution our RAM24C2 extract formulation hastens the cure of conjunctivitis developed rabbits and need less dosage frequency.

  3. 红树林海洋淤泥中放线菌的分离与鉴定%Identification and Analysis of Actinomycetes in Marine Mud of Mangrove

    Institute of Scientific and Technical Information of China (English)

    陈森洲; 刘菁; 陈建宏; 骆耐香; 钟毓娟; 蒋莲秀

    2011-01-01

    To explore actinomycetes resources in marine mud of mangrove in Beihai city of Guangxi province,actinomycete samples in marine mud of mangrove were separated by using seawater prepared Gause culture medium, total DNA of 10 typical strains of actinomycetes was screened, separated and extracted, 16Sr DNA was amplified by PCR with universal primers. The results from DNA sequencing and comparison identification of amplified product show that 10 typical strains of actinomycetes belong to 2 genera, 8 of which are common actinomycetes and belong to Streptomyces (80%), the other 2 are rare actinomycetes and belong to Nocardia (20%). All above indicate that there are abundant species of actinomycetes in marine mud of mangrove in Beihai city of Guangxi province.%为了解广西北海红树林海洋淤泥中的放线菌资源,采用海水配制高氏培养基分离红树林海洋淤泥中的放线菌样品,从中筛选、分离、提取lO株典型放线菌菌株总DNA,用放线菌通用引物对16Sr DNA进行PCR扩增,对获得的扩增结果进行DNA序列测定和对比鉴定.结果表明,10株典型放线菌菌株为2种菌属,其中有8株为链霉菌属(80%),为常见放线菌;有2株为拟诺卡氏菌属(20%),为稀有放线菌.表明,广西北海红树林海洋淤泥蕴含着种类丰富的放线菌.

  4. Inhibition of Aspergillus parasiticus and cancer cells by marine actinomycete strains

    Science.gov (United States)

    Li, Ping; Yan, Peisheng

    2014-12-01

    Ten actinomycete strains isolated from the Yellow Sea off China's coasts were identified as belonging to two genera by 16S rDNA phylogenetic analysis: Streptomyces and Nocardiopsis. Six Streptomyces strains (MA10, 2SHXF01-3, MA35, MA05-2, MA05-2-1 and MA08-1) and one Nocardiopsis strain (MA03) were predicted to have the potential to produce aromatic polyketides based on the analysis of the KSα (ketoacyl-synthase) gene in the type II PKS (polyketides synthase) gene cluster. Four strains (MA03, MA01, MA10 and MA05-2) exhibited significant inhibitory effects on mycelia growth (inhibition rate >50%) and subsequent aflatoxin production (inhibition rate >75%) of the mutant aflatoxigenic Aspergillus parasiticus NFRI-95. The ethyl acetate extracts of the broth of these four strains displayed significant inhibitory effects on mycelia growth, and the IC50 values were calculated (MA03: 0.275 mg mL-1, MA01: 0.106 mg mL-1, MA10: 1.345 mg mL-1 and MA05-2: 1.362 mg mL-1). Five strains (2SHXF01-3, MA03, MA05-2, MA01 and MA08-1) were selected based on their high cytotoxic activities. The ethyl acetate extract of the Nocardiopsis strain MA03 was particularly noted for its high antitumor activity against human carcinomas of the cervix (HeLa), lung (A549), kidney (Caki-1) and liver (HepG2) (IC50: 2.890, 1.981, 3.032 and 2.603 μg mL-1, respectively). The extract also remarkably inhibited colony formation of HeLa cells at an extremely low concentration (0.5 μg mL-1). This study highlights that marine-derived actinomycetes are a huge resource of compounds for the biological control of aflatoxin contamination and the development of novel drugs for human carcinomas.

  5. Isolation and Identification of a Rare Actinomycete with Antibacterial Activity from Saline Region of Iran

    Directory of Open Access Journals (Sweden)

    Samaneh Mashhadi

    2016-07-01

    Full Text Available Background: The appearance of multi-drug resistant microorganisms is becoming a global problem. Already several strategies have been employed to overcome antibiotic resistance issue. Developing new antimicrobial compounds from microbial sources could be a beneficial solution. Hence screening programs in order to discover new antibiotics from microbial entities are interesting. Because of high capabilities of extremophiles for adaptation to harsh environmental conditions, the microbial communities of the extreme environments could be regarded as rich resources for new antibacterial metabolites. Materials and Methods: In this research different saline environments of Iran have been subjected to screening of antibiotic producing actinomycetes using overlaid method after the ingredient optimization of culture media. The strain which was shown pronounce inhibition zone in the screening step, has been phylogenetically analyzed followed by studying the effect of agar concentration and cultivation time on the production of antibacterial agent(s. Results: The strain RS1, a rare actinomycete, had antibacterial activity against Escherichia coli (PTCC 1330 and Bacillus subtilis (PTCC 1023 and taxonomically belongs to the genus Amycolatopsis with high similarity of 99.6% to Amycolatopsis coloradensis IMSNU 22096T based on sequencing of 16S rRNA gene nucleotide. The zone of growth inhibition of E.coli was the widest when the base layer had contained 1.2% agar, while no significant differences were observed on anti-gram-positive bacterial assay. This strain produced the antibacterial agent at the highest level after 5 days when B. subtilis was used as an indicator, but the production of antibacterial agent active against E.Coli was reached to its highest level on the 3rd days of cultivation and then was decreased significantly. Conclusion: Due to the results of agar concentration and time course study as well as possessing activity against both Gram

  6. Identification of actinomycetes from plant rhizospheric soils with inhibitory activity against Colletotrichum spp., the causative agent of anthracnose disease.

    Science.gov (United States)

    Intra, Bungonsiri; Mungsuntisuk, Isada; Nihira, Takuya; Igarashi, Yasuhiro; Panbangred, Watanalai

    2011-04-01

    Colletotrichum is one of the most widespread and important genus of plant pathogenic fungi worldwide. Various species of Colletotrichum are the causative agents of anthracnose disease in plants, which is a severe problem to agricultural crops particularly in Thailand. These phytopathogens are usually controlled using chemicals; however, the use of these agents can lead to environmental pollution. Potential non-chemical control strategies for anthracnose disease include the use of bacteria capable of producing anti-fungal compounds such as actinomycetes spp., that comprise a large group of filamentous, Gram positive bacteria from soil. The aim of this study was to isolate actinomycetes capable of inhibiting the growth of Colletotrichum spp, and to analyze the diversity of actinomycetes from plant rhizospheric soil. A total of 304 actinomycetes were isolated and tested for their inhibitory activity against Colletotrichum gloeosporioides strains DoA d0762 and DoA c1060 and Colletotrichum capsici strain DoA c1511 which cause anthracnose disease as well as the non-pathogenic Saccharomyces cerevisiae strain IFO 10217. Most isolates (222 out of 304, 73.0%) were active against at least one indicator fungus or yeast. Fifty four (17.8%) were active against three anthracnose fungi and 17 (5.6%) could inhibit the growth of all three fungi and S. cerevisiae used in the test. Detailed analysis on 30 selected isolates from an orchard at Chanthaburi using the comparison of 16S rRNA gene sequences revealed that most of the isolates (87%) belong to the genus Streptomyces sp., while one each belongs to Saccharopolyspora (strain SB-2) and Nocardiopsis (strain CM-2) and two to Nocardia (strains BP-3 and LK-1). Strains LC-1, LC-4, JF-1, SC-1 and MG-1 exerted high inhibitory activity against all three anthracnose fungi and yeast. In addition, the organic solvent extracts prepared from these five strains inhibited conidial growth of the three indicator fungi. Preliminary analysis of crude

  7. Screening of phospholipase A activity and its production by new actinomycete strains cultivated by solid-state fermentation

    Directory of Open Access Journals (Sweden)

    Priscila Sutto-Ortiz

    2017-07-01

    Full Text Available Novel microbial phospholipases A (PLAs can be found in actinomycetes which have been poorly explored as producers of this activity. To investigate microbial PLA production, efficient methods are necessary such as high-throughput screening (HTS assays for direct search of PLAs in microbial cultures and cultivation conditions to promote this activity. About 200 strains isolated with selected media for actinomycetes and mostly belonging to Streptomyces (73% and Micromonospora (10% genus were first screened on agar-plates containing the fluorophore rhodamine 6G and egg yolk phosphatidylcholine (PC to detect strains producing phospholipase activity. Then, a colorimetric HTS assay for general PLA activity detection (cHTS-PLA using enriched PC (≈60% as substrate and cresol red as indicator was developed and applied; this cHTS-PLA assay was validated with known PLAs. For the first time, actinomycete strains were cultivated by solid-state fermentation (SSF using PC as inductor and sugar-cane bagasse as support to produce high PLA activity (from 207 to 2,591 mU/g of support. Phospholipase activity of the enzymatic extracts from SSF was determined using the implemented cHTS-PLA assay and the PC hydrolysis products obtained, were analyzed by TLC showing the presence of lyso-PC. Three actinomycete strains of the Streptomyces genus that stood out for high accumulation of lyso-PC, were selected and analyzed with the specific substrate 1,2-α-eleostearoyl-sn-glycero-3-phosphocholine (EEPC in order to confirm the presence of PLA activity in their enzymatic extracts. Overall, the results obtained pave the way toward the HTS of PLA activity in crude microbial enzymatic extracts at a larger scale. The cHTS-PLA assay developed here can be also proposed as a routine assay for PLA activity determination during enzyme purification,directed evolution or mutagenesis approaches. In addition, the production of PLA activity by actinomycetes using SSF allow find and

  8. Actinomadura gamaensis sp. nov., a novel actinomycete isolated from soil in Gama, Chad.

    Science.gov (United States)

    Abagana, Adam Yacoub; Sun, Pengyu; Liu, Chongxi; Cao, Tingting; Zheng, Weiwei; Zhao, Shanshan; Xiang, Wensheng; Wang, Xiangjing

    2016-06-01

    A novel single spore-producing actinomycete, designated strain NEAU-Gz5(T), was isolated from a soil sample from Gama, Chad. A polyphasic taxonomic study was carried out to establish the status of this strain. The diamino acid present in the cell wall is meso-diaminopimelic acid. Glucose, mannose and madurose occur in whole cell hydrolysates. The polar lipids were found to consist of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol mannoside and an unidentified glycolipid. The predominant menaquinones were identified as MK-9(H8) and MK-9(H6). The predominant cellular fatty acids were found to be C16:0, iso-C15:0, iso-C16:0 and C18:0 10-methyl. Phylogenetic analysis based on the 16S rRNA gene showed that strain NEAU-Gz5(T) belongs to the genus Actinomadura and is closely related to Actinomadura oligospora JCM 10648(T) (ATCC 43269(T); 98.3 % similarity). However, the low level of DNA-DNA relatedness and some different phenotypic characteristics allowed the strain to be distinguished from its close relatives. Therefore, it is concluded that strain NEAU-Gz5(T) represents a novel species of the genus of Actinomadura, for which the name Actinomadura gamaensis sp. nov. is proposed. The type strain is NEAU-Gz5(T) (= CGMCC 4.7301(T) = DSM 100815(T)).

  9. Diversity and Bioactivity of Actinomycetes from Marine Sediments of the Yellow Sea

    Institute of Scientific and Technical Information of China (English)

    ZHANG Shumin; YE Liang; TANG Xuexi

    2012-01-01

    Among the 116 actinomycetes collected from marine sediments of the Yellow Sea,56 grew slowly and appeared after 2-3 weeks of incubation.Among the 56 strains,only 3 required seawater (SW) for growth,and 21 grew well in the medium prepared with SW rather than distilled water (DW),while the remaining 32 grew well either with SW or with DW.Six representatives with different morphological characteristics,including 1 SW-requiring strain and 5 well-growing with SW strains,were selected for phylogenetic analysis based on 16S rRNA gene.Two strains belong to Micrococcaceae and Nocardiopsaceae respectively.The other 4strains belong to the family of Streptomycetaceae.In the analyzed 6 strains,one was related to Nocardiopsis spp.and the other three were related to Streptomyces spp.,representing new taxa.Bioactivity testing of fermentation products from 3 SW-requiring strains and 21 well-growing with SW strains revealed that 17 strains possessed remarkable activities against gram-positive pathogen or/and tumor cells,suggesting that they were prolific resources for natural drug discovery.

  10. A method to type the potential angucycline producers in actinomycetes isolated from marine sponges.

    Science.gov (United States)

    Ouyang, Yongchang; Wu, Houbo; Xie, Lianwu; Wang, Guanghua; Dai, Shikun; Chen, Minjie; Yang, Keqian; Li, Xiang

    2011-05-01

    Angucyclines are aromatic polyketides with antimicrobial, antitumor, antiviral and enzyme inhibition activities. In this study, a new pair of degenerate primers targeting the cyclase genes that are involved in the aromatization of the first and/or second ring of angucycline, were designed and evaluated in a PCR protocol targeting the jadomycin cyclase gene of Streptomyces venezuelae ISP5230. The identity of the target amplicon was confirmed by sequencing. After validation, the primers were used to screen 49 actinomycete isolates from three different marine sponges to identify putative angucycline producers. Seven isolates were positively identified using this method. Sequence analysis of the positive amplicons confirmed their identity as putative angucycline cyclases with sequence highly similar to known angucycline cyclases. Phylogenetic analysis clustered these positives into the angucycline group of cyclases. Furthermore, amplifications of the seven isolates using ketosynthase-specific primers were positive, backing the results using the cyclase primers. Together these results provided strong support for the presence of angucycline biosynthetic genes in these isolates. The specific primer set targeting the cyclase can be used to identify putative angucycline producers among marine actinobacteria, and aid in the discovery of novel angucyclines.

  11. Effects of marine actinomycete on the removal of a toxicity alga Phaeocystis globose in eutrophication waters

    Directory of Open Access Journals (Sweden)

    Huajun eZhang

    2015-05-01

    Full Text Available Phaeocystis globosa blooms in eutrophication waters can cause severely damage in marine ecosystem and consequently influence human activities. This study investigated the effect and role of an algicidal actinomycete (Streptomyces sp. JS01 on the elimination process of P. globosa. JS01 supernatant could alter algal cell membrane permeability in 4 h when analyzed with flow cytometry. Reactive oxygen species (ROS levels were 7.2 times higher than that at 0 h following exposure to JS01 supernatant for 8 h, which indicated that algal cells suffered from oxidative damage. The Fv/Fm value which could reflect photosystem II (PS II electron flow status also decreased. Real-time PCR showed that the expression of the photosynthesis related genes psbA and rbcS were suppressed by JS01 supernatant, which might induce damage to PS II. Our results demonstrated that JS01 supernatant can change algal membrane permeability in a short time and then affect photosynthesis process, which might block the PS II electron transport chain to produce excessive ROS. This experiment demonstrated that Streptomyces sp. JS01 could eliminate harmful algae in marine waters efficiently and may be function as a harmful algal bloom controller material.

  12. Streptomyces alkalithermotolerans sp. nov., a novel alkaliphilic and thermotolerant actinomycete isolated from a soda lake.

    Science.gov (United States)

    Sultanpuram, Vishnuvardhan Reddy; Mothe, Thirumala; Mohammed, Farooq

    2015-02-01

    An alkaliphilic actinomycete, strain AC3(T), was isolated from Lonar soda lake, in India. Based on 16S rRNA gene sequence analysis it was identified that the strain belongs to the class Actinobacteria and was most closely related to Streptomyces sodiiphilus JCM 13581(T) (96.4 % sequence similarity), Streptomyces leeuwenhoekii DSM 42122(T) (96.1 %), Streptomyces albus NRRL B-2365(T) (96.1 %), Streptomyces panacagri Gsoil 519(T) (96.0 %), Streptomyces fimbriatus NBRC 15411(T) (95.9 %) and other members of the genus Streptomyces (cream substrate and white aerial mycelia on most tested media. The optimum pH for growth was determined to be 9.5-10.0 with no growth at pH 7.0. The DNA G+C content of strain AC3(T) was determined to be 71.2 mol %. The results of the polyphasic analysis allowed a clear differentiation of strain AC3(T) from all other members of the genus Streptomyces. Strain AC3(T) is thus considered to represent a novel member of the genus Streptomyces, for which the name Streptomyces alkalithermotolerans sp. nov. is proposed. The type strain is AC3(T) (=KCTC 29497(T) = JCM 30167(T)).

  13. Generic and functional diversity in endophytic actinomycetes from wild Compositae plant species at South Sinai - Egypt.

    Science.gov (United States)

    El-Shatoury, Sahar A; El-Kraly, Omnia A; Trujillo, Martha E; El-Kazzaz, Waleed M; El-Din, El-Sayeda Gamal; Dewedar, Ahmed

    2013-09-01

    The diversity of culturable endophytic actinomycetes associated with wild Compositae plants is scantily explored. In this study, one hundred and thirty one endophytic actinobacteria were isolated from ten Compositae plant species collected from South Sinai in Egypt. Microscopic and chemotaxonomic investigation of the isolates indicated fourteen genera. Rare genera, such as Microtetraspora, and Intrasporangium, which have never been previously reported to be endophytic, were identified. Each plant species accommodated between three to eight genera of actinobacteria and unidentified strains were recovered from seven plant species. The generic diversity analysis of endophytic assemblages grouped the plant species into three main clusters, representing high, moderate and low endophytic diversity. The endophytes showed high functional diversity, based on forty four catabolic and plant growth promotion traits; providing some evidence that such traits could represent key criteria for successful residence of endophytes in the endosphere. Stress-tolerance traits were more predictive measure of functional diversity differences between the endophyte assemblages (Shannon's index, p = 0.01). The results indicate a potential prominent role of endophytes for their hosts and emphasize the potency of plant endosphere as a habitat for actinobacteria with promising future applications.

  14. [Actinomycosic mycetoma of the foot in Morocco due to Actinomycetes viscosus].

    Science.gov (United States)

    Baha, H; Khadir, K; Hali, F; Benchikhi, H; Zeghwagh, A; Zerouali, K; Belabbes, H; El Mdaghri, N; Soussi, M A; Marnissi, F; Kadioui, F

    2015-03-01

    We present the case of an actinomycotic mycetoma of the foot due to Actinomycetes viscosus. It evolved for nine years on the foot of a 26-year-old patient from a rural environment: Douar Inezgane (city in southern Morocco). Bacteriological study of the skin and grains confirmed the diagnosis. It showed positive bacilli on direct examination and on Gram staining and in positive culture. Histological study showed a polymorphous granulomatous inflammation without signs of malignancy with actinomycotic grains. Then we retained the diagnosis of primary cutaneous actinomycosis without visceral locations. The treatment was based on antibiotics: penicillin G by intravenous infusion for five weeks, relayed orally by amoxicillin associated with trimethoprim-sulfamethoxazole for long periods. After six months of treatment, we observed a favorable outcome with reduction of the swelling, nodules, lymphadenopathy, fistula's number and extension of time of issue of grains. The current follow up is 15 months. The primary cutaneous actinomycosis is still relevant in Morocco. Copyright © 2015. Published by Elsevier Masson SAS.

  15. Amycolatopsis thailandensis sp. nov., a poly(L-lactic acid)-degrading actinomycete, isolated from soil.

    Science.gov (United States)

    Chomchoei, Atchareeya; Pathom-Aree, Wasu; Yokota, Akira; Kanongnuch, Chartchai; Lumyong, Saisamorn

    2011-04-01

    A novel actinomycete that was capable of degrading poly(l-lactic acid), strain CMU-PLA07(T), was isolated from soil in northern Thailand. Strain CMU-PLA07(T) had biochemical, chemotaxonomic, morphological and physiological properties that were consistent with its classification in the genus Amycolatopsis. 16S rRNA gene sequence analysis showed that the isolate formed a phyletic line within the genus Amycolatopsis. Strain CMU-PLA07(T) was most similar to Amycolatopsis coloradensis IMSNU 22096(T) (99.5 % 16S rRNA gene sequence similarity) and Amycolatopsis alba DSM 44262(T) (99.4 %). However, strain CMU-PLA07(T) was distinguishable from the type strains of species of the genus Amycolatopsis on the basis of DNA-DNA relatedness and phenotypic data. Therefore, strain CMU-PLA07(T) is considered to represent a novel species of the genus Amycolatopsis, for which the name Amycolatopsis thailandensis sp. nov. is proposed. The type strain is CMU-PLA07(T) ( = JCM 16380(T) = BCC 38279(T)).

  16. Biotechnological potential of endophytic actinomycetes associated with Asteraceae plants: isolation, biodiversity and bioactivities.

    Science.gov (United States)

    Tanvir, Rabia; Sajid, Imran; Hasnain, Shahida

    2014-04-01

    Endophytic actinomycetes from five Asteraceae plants were isolated and evaluated for their bioactivities. From Parthenium hysterophorus, Ageratum conyzoides, Sonchus oleraceus, Sonchus asper and Hieracium canadense, 42, 45, 90, 3, and 2 isolates, respectively, were obtained. Of the isolates, 86 (47.2 %) showed antimicrobial activity. Majority of the isolates were recovered from the roots (n = 127, 69.7 %). The dominant genus was Streptomyces (n = 96, 52.7 %), while Amycolatopsis, Pseudonocardia, Nocardia and Micromonospora were also recovered. Overall, 36 of the 86 isolates were significantly bioactivity while 18 (20.9 %) showed strong bioactivity. In total, 52.1 and 66.6 % showed potent cytotoxicity and antioxidant activities. The LC50 for 15 strains was <20 μg/ml. Compared to the ascorbate standard (EC50 0.34 μg/ml), all isolates gave impressive results with notable EC50 values of 0.65, 0.67, 0.74 and 0.79 μg/ml.

  17. Conjugal transfer of a virulence plasmid in the opportunistic intracellular actinomycete Rhodococcus equi.

    Science.gov (United States)

    Tripathi, V N; Harding, W C; Willingham-Lane, J M; Hondalus, M K

    2012-12-01

    Rhodococcus equi is a facultative intracellular, Gram-positive, soilborne actinomycete which can cause severe pyogranulomatous pneumonia with abscessation in young horses (foals) and in immunocompromised people, such as persons with AIDS. All strains of R. equi isolated from foals and approximately a third isolated from humans contain a large, ~81-kb plasmid which is essential for the intramacrophage growth of the organism and for virulence in foals and murine in vivo model systems. We found that the entire virulence plasmid could be transferred from plasmid-containing strains of R. equi (donor) to plasmid-free R. equi strains (recipient) at a high frequency and that plasmid transmission reestablished the capacity for intracellular growth in macrophages. Plasmid transfer required living cells and cell-to-cell contact and was unaffected by the presence of DNase, factors pointing to conjugation as the major means of genetic transfer. Deletion of a putative relaxase-encoding gene, traA, located in the proposed conjugative region of the plasmid, abolished plasmid transfer. Reversion of the traA mutation restored plasmid transmissibility. Finally, plasmid transmission to other Rhodococcus species and some additional related organisms was demonstrated. This is the first study showing a virulence plasmid transfer in R. equi, and it establishes a mechanism by which the virulence plasmid can move among bacteria in the soil.

  18. Marine actinomycete crude extracts with potent TRAIL-resistance overcoming activity against breast cancer cells.

    Science.gov (United States)

    Elmallah, Mohammed I Y; Micheau, Olivier; Eid, Mennat Allah G; Hebishy, Ali M S; Abdelfattah, Mohamed S

    2017-06-01

    Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising anticancer agent, as it can kill tumor cells selectively. In our search of bioactive natural products to overcome TRAIL-resistance, we isolated 47 actinomycete strains from different sediments and seawater samples collected from the Red Sea coast in Egypt and found four crude extracts (EGY1, EGY3, EGY24 and EGY34) displaying TRAIL sensitizing activity in the resistant breast cancer cell line MDA-MB-231. None of these crude extracts exhibited cytotoxic effect on normal mouse embryonic fibroblasts (MEF), with the exception of EGY34. Analysis of the signaling pathways underlying the sensitization of MDA-MB-231 cells to TRAIL-induced apoptosis, by western blotting, revealed that all crude extracts facilitated initiator caspase‑8/-10 activation upon TRAIL stimulation, but that in addition, EGY3 and EGY34, alone, induced strong ER-stress activation, with the appearance of BiP in the cytosolic extracts. Our results pave the way to the discovery and the development of marine-derived drugs for cancer therapy.

  19. Antibiotic-producing ability by representatives of a newly discovered lineage of actinomycetes.

    Science.gov (United States)

    Busti, Elena; Monciardini, Paolo; Cavaletti, Linda; Bamonte, Ruggiero; Lazzarini, Ameriga; Sosio, Margherita; Donadio, Stefano

    2006-03-01

    The discovery of new antibiotics and other bioactive microbial metabolites continues to be an important objective in new drug research. Since extensive screening has led to the discovery of thousands of bioactive microbial molecules, new approaches must be taken in order to reduce the probability of rediscovering known compounds. The authors have recently isolated slow-growing acidophiles belonging to the novel genera Catenulispora and Actinospica within the order Actinomycetales. These strains, which likely belong to a new suborder, grow as filamentous mycelia, have a genome size around 8 Mb, and produce antimicrobial activities. In addition, a single strain harbours simultaneously genes encoding type I and type II polyeketide synthases, as well as non-ribosomal peptide synthetases. The metabolite produced by one strain was identified as a previously reported dimeric isochromanequinone. In addition, at least the Catenulispora strains appear globally distributed, since a PCR-specific signal could be detected in a significant fraction of acidic soils from different continents, and similar strains have been independently isolated from an Australian soil (Jospeh et al., Appl Environ Microbiol 69, 7210-7215, 2003). Thus, these previously uncultured actinomycetes share several features with Streptomyces and related antibiotic-producing genera, and represent a promising source of novel antibiotics.

  20. Streptomyces castaneus sp. nov., a novel actinomycete isolated from the rhizosphere of Peucedanum praeruptorum Dunn.

    Science.gov (United States)

    Zhou, Shuyu; Li, Zhilei; Bai, Lu; Yan, Kai; Zhao, Junwei; Lu, Chang; Liu, Chongxi; Wang, Xiangjing; Xiang, Wensheng

    2017-01-01

    During an investigation of microbial diversity in medicinal herbs, a novel actinomycete, strain NEAU-QHHV11(T) was isolated from the rhizosphere of Peucedanum praeruptorum Dunn collected from Xianglu Mountain in Heilongjiang Province, northeast China and characterized using a polyphasic approach. The organism was found to have typical characteristics of the genus Streptomyces. Phylogenetic analysis based on 16S rRNA gene sequence also indicated that strain NEAU-QHHV11(T) belongs to the genus Streptomyces and was most closely related to Streptomyces graminilatus NBRC 108882(T) (98.7 % sequence similarity) and Streptomyces turgidiscabies NBRC 16080(T) (98.7 % sequence similarity). The results of DNA-DNA hybridization and some phenotypic characteristics indicated that strain NEAU-QHHV11(T) could be distinguished from its close phylogenetic relatives. Thus, strain NEAU-QHHV11(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces castaneus sp. nov. is proposed. The type strain is NEAU-QHHV11(T) (=CGMCC 4.7235(T) = DSM 100520(T)).

  1. Cloning and characterization of the first actinomycete β-propeller phytase from Streptomyces sp. US42.

    Science.gov (United States)

    Boukhris, Ines; Farhat-Khemakhem, Ameny; Bouchaala, Kameleddine; Virolle, Marie-Joëlle; Chouayekh, Hichem

    2016-10-01

    A gene encoding an extracellular phytase was cloned for the first time from an Actinomycete, Streptomyces sp. US42 and sequenced. The sequence of this gene revealed an encoded polypeptide (PHY US42) exhibiting one and six residues difference with the putative phytases of Streptomyces lividans TK24 and Streptomyces coelicolor A3(2), respectively. The molecular modeling of PHY US42 indicated that this phytase belongs to the group of β-propeller phytases that are usually calcium-dependent. PHY US42 was purified and characterized. Its activity was calcium-dependent and maximal at pH 7 and 65 °C. The enzyme was perfectly stable at pH ranging from 5 to 10 and its thermostability was greatly enhanced in the presence of calcium. Indeed, PHY US42 maintained 80% of activity after 10 min of incubation at 75 °C in the presence of 5 mM CaCl2 . PHY US42 was also found to exhibit high stability after incubation at 37 °C for 1 h in the presence of bovine bile and digestive proteases like of pepsin, trypsin, and chymotrypsin. Considering its biochemical properties, PHY US42 could be used as feed additive in combination with an acid phytase for monogastric animals.

  2. Streptomyces gamaensis sp. nov., a novel actinomycete with antifungal activity isolated from soil in Gama, Chad.

    Science.gov (United States)

    Zhao, Shanshan; Ye, Lan; Liu, Chongxi; Abagana, Adam Yacoub; Zheng, Weiwei; Sun, Pengyu; Li, Jiansong; Xiang, Wensheng; Wang, Xiangjing

    2017-04-01

    During an investigation exploring potential sources of novel species and natural products, a novel actinomycete with antifungal activity, designated strain NEAU-Gz11(T), was isolated from a soil sample, which was collected from Gama, Chad. The isolate was found to have morphological and chemotaxonomic characteristics typical of members of the genus Streptomyces. 16S rRNA gene sequence similarity studies showed that strain NEAU-Gz11(T) belongs to the genus Streptomyces with high sequence similarity to Streptomyces hiroshimensis JCM 4098(T) (98.0 %). Similarities to other type strains of the genus Streptomyces were lower than 98.0 %. However, the physiological and biochemical characteristics and low levels of DNA-DNA relatedness could differentiate the isolate genotypically and phenotypically from S. hiroshimensis JCM 4098(T). Therefore, the strain is concluded to represent a novel species of the genus Streptomyces, for which the name Streptomyces gamaensis sp. nov. is proposed. The type strain is NEAU-Gz11(T) (=CGMCC 4.7304(T)=DSM 101531(T)).

  3. Streptomyces atlanticus sp. nov., a novel actinomycete isolated from marine sponge Aplysina fulva (Pallas, 1766).

    Science.gov (United States)

    Silva, Fábio Sérgio Paulino; Souza, Danilo Tosta; Zucchi, Tiago Domingues; Pansa, Camila Cristiane; de Figueiredo Vasconcellos, Rafael Leandro; Crevelin, Eduardo José; de Moraes, Luiz Alberto Beraldo; Melo, Itamar Soares

    2016-11-01

    The taxonomic position of a novel marine actinomycete isolated from a marine sponge, Aplysina fulva, which had been collected in the Archipelago of Saint Peter and Saint Paul (Equatorial Atlantic Ocean), was determined by using a polyphasic approach. The organism showed a combination of morphological and chemotaxonomic characteristics consistent with its classification in the genus Streptomyces and forms a distinct branch within the Streptomyces somaliensis 16S rRNA gene tree subclade. It is closely related to Streptomyces violascens ISP 5183(T) (97.27 % 16S rRNA gene sequence similarity) and Streptomyces hydrogenans NBRC 13475(T) (97.15 % 16S rRNA gene sequence similarity). The 16S rRNA gene similarities between the isolate and the remaining members of the subclade are lower than 96.77 %. The organism can be distinguished readily from other members of the S. violacens subclade using a combination of phenotypic properties. On the basis of these results, it is proposed that isolate 103(T) (=NRRL B-65309(T) = CMAA 1378(T)) merits recognition as the type strain of a new Streptomyces species, namely Streptomyces atlanticus sp. nov.

  4. Biofilm formation and partial biodegradation of polystyrene by the actinomycete Rhodococcus ruber: biodegradation of polystyrene.

    Science.gov (United States)

    Mor, Roi; Sivan, Alex

    2008-11-01

    Polystyrene, which is one of the most utilized thermoplastics, is highly durable and is considered to be non-biodegradable. Hence, polystyrene waste accumulates in the environment posing an increasing ecological threat. In a previous study we have isolated a biofilm-producing strain (C208) of the actinomycete Rhodococcus ruber that degraded polyethylene films. Formation of biofilm, by C208, improved the biodegradation of polyethylene. Consequently, the present study aimed at monitoring the kinetics of biofilm formation by C208 on polystyrene, determining the physiological activity of the biofilm and analyzing its capacity to degrade polystyrene. Quantification of the biofilm biomass was performed using a modified crystal violet (CV) staining or by monitoring the protein content in the biofilm. When cultured on polystyrene flakes, most of the bacterial cells adhered to the polystyrene surface within few hours, forming a biofilm. The growth of the on polystyrene showed a pattern similar to that of a planktonic culture. Furthermore, the respiration rate, of the biofilm, exhibited a pattern similar to that of the biofilm growth. In contrast, the respiration activity of the planktonic population showed a constant decline with time. Addition of mineral oil (0.005% w/v), but not non-ionic surfactants, increased the biofilm biomass. Extended incubation of the biofilm for up to 8 weeks resulted in a small reduction in the polystyrene weight (0.8% of gravimetric weight loss). This study demonstrates the high affinity of C208 to polystyrene which lead to biofilm formation and, presumably, induced partial biodegradation.

  5. Nocardiopsis arabia sp. nov., a halotolerant actinomycete isolated from a sand-dune soil.

    Science.gov (United States)

    Hozzein, Wael N; Goodfellow, Michael

    2008-11-01

    The taxonomic status of an unknown actinomycete isolated from a sand-dune soil was established using a polyphasic approach. Isolate S186(T) had chemotaxonomic and morphological properties consistent with its classification in the genus Nocardiopsis, grew on agar plates at NaCl concentrations of up to 15 % (w/v) and formed a distinct phyletic line in the Nocardiopsis 16S rRNA gene sequence tree. Its closest phylogenetic neighbours were Nocardiopsis chromatogenes, Nocardiopsis composta, Nocardiopsis gilva and Nocardiopsis trehalosi, with sequence similarity to the various type strains of 96.9 %, but it was readily distinguished from the type strains of these and related species using a range of phenotypic properties. It is apparent from the genotypic and phenotypic data that strain S186(T) belongs to a novel species of the genus Nocardiopsis, for which the name Nocardiopsis arabia sp. nov. is proposed. The type strain is S186(T) (=CGMCC 4.2057(T) =DSM 45083(T)).

  6. Trehalose lipid biosurfactants produced by the actinomycetes Tsukamurella spumae and T. pseudospumae.

    Science.gov (United States)

    Kügler, Johannes H; Muhle-Goll, Claudia; Kühl, Boris; Kraft, Axel; Heinzler, Raphael; Kirschhöfer, Frank; Henkel, Marius; Wray, Victor; Luy, Burkhard; Brenner-Weiss, Gerald; Lang, Siegmund; Syldatk, Christoph; Hausmann, Rudolf

    2014-11-01

    Actinomycetales are known to produce various secondary metabolites including products with surface-active and emulsifying properties known as biosurfactants. In this study, the nonpathogenic actinomycetes Tsukamurella spumae and Tsukamurella pseudospumae are described as producers of extracellular trehalose lipid biosurfactants when grown on sunflower oil or its main component glyceryltrioleate. Crude extracts of the trehalose lipids were purified using silica gel chromatography. The structure of the two trehalose lipid components (TL A and TL B) was elucidated using a combination of matrix-assisted laser desorption/ionization time-of-flight/time-of-flight/tandem mass spectroscopy (MALDI-ToF-ToF/MS/MS) and multidimensional NMR experiments. The biosurfactants were identified as 1-α-glucopyranosyl-1-α-glucopyranosid carrying two acyl chains varying of C4 to C6 and C16 to C18 at the 2' and 3' carbon atom of one sugar unit. The trehalose lipids produced demonstrate surface-active behavior and emulsifying capacity. Classified as risk group 1 organisms, T. spumae and T. pseudospumae hold potential for the production of environmentally friendly surfactants.

  7. Diversity and bioactivity of actinomycetes from marine sediments of the Yellow Sea

    Science.gov (United States)

    Zhang, Shumin; Ye, Liang; Tang, Xuexi

    2012-03-01

    Among the 116 actinomycetes collected from marine sediments of the Yellow Sea, 56 grew slowly and appeared after 2-3 weeks of incubation. Among the 56 strains, only 3 required seawater (SW) for growth, and 21 grew well in the medium prepared with SW rather than distilled water (DW), while the remaining 32 grew well either with SW or with DW. Six representatives with different morphological characteristics, including 1 SW-requiring strain and 5 well-growing with SW strains, were selected for phylogenetic analysis based on 16S rRNA gene. Two strains belong to Micrococcaceae and Nocardiopsaceae respectively. The other 4 strains belong to the family of Streptomycetaceae. In the analyzed 6 strains, one was related to Nocardiopsis spp. and the other three were related to Streptomyces spp., representing new taxa. Bioactivity testing of fermentation products from 3 SW-requiring strains and 21 well-growing with SW strains revealed that 17 strains possessed remarkable activities against gram-positive pathogen or/and tumor cells, suggesting that they were prolific resources for natural drug discovery.

  8. Antibiotic pigment from desert soil actinomycetes; biological activity, purification and chemical screening

    Directory of Open Access Journals (Sweden)

    Selvameenal L

    2009-01-01

    Full Text Available An actinomycete strain, Streptomyces hygroscopicus subsp. ossamyceticus (strain D10 was isolated from Thar Desert soil, Rajasthan during the year 2006 and found to produce a yellow color pigment with antibiotic activity. Crude pigment was produced from strain D10 by solid state fermentation using wheat bran medium followed by extraction with ethyl acetate. The antimicrobial activity of the crude pigment was evaluated against drug resistant pathogens such as methicillin-resistant Staphylococcus aureus, vancomycin-resistant Staphylococcus aureus, extended spectrum b-lactamase producing cultures of Escherichia coli, Pseudomonas aeruginosa and Klebsiella sp. About 420 mg of crude pigment was produced per 10 g of wheat bran medium. In the disc diffusion method the crude ethyl acetate extract showed a minimum of 10 mm inhibition against Klebsiella sp. and maximum of 19 mm of inhibition against Escherichia coli. The crude pigment was partially purified using thin layer chromatography with the solvent system chloroform:methanol (30:70 and the Rf value was calculated as 0.768. Antimicrobial activity of the partially purified compound from thin layer chromatography was determined using the bioautography method. The purified pigment showed minimum of 15 mm inhibition against Klebsiella sp. and a maximum of 23 mm of inhibition against vancomycin-resistant Staphylococcus aureus in the disc diffusion method. Based on the results of chemical screening, the pigment was tentatively identified as group of sugar containing molecules.

  9. Actinomycetospora iriomotensis sp. nov., a novel actinomycete isolated from a lichen sample.

    Science.gov (United States)

    Yamamura, Hideki; Ashizawa, Haruna; Nakagawa, Youji; Hamada, Moriyuki; Ishida, Yuumi; Otoguro, Misa; Tamura, Tomohiko; Hayakawa, Masayuki

    2011-04-01

    An actinomycete strain, IR73-Li102(T), was isolated from a lichen sample obtained from Iriomote Island, Japan, and subsequently characterized using a polyphasic approach. Comparative 16S rRNA gene sequence analysis revealed that strain IR73-Li102(T) had the highest sequence similarities with Actinomycetospora chiangmaiensis YIM 0006(T) (98.3%), A. corticola 014-5(T) (98.1%) and A. rishiriensis RI109-Li102(T) (98.0%). However, DNA-DNA hybridization assays, as well as physiological and biochemical analyzes, showed that strain IR73-Li102(T) could be clearly differentiated from its closest phylogenetic relatives. The strain contained meso-diaminopimelic acid, and arabinose and galactose were present in whole-cell hydrolysates. The predominant menaquinone was MK-8(H(4)), and the diagnostic phospholipids were phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol and diphosphatidylglycerol. The major cellular fatty acid was iso-C(16:0) (58%). The chemotaxonomic properties of strain IR73-Li102(T) were consistent with those shared by members of the genus Actinomycetospora. On the basis of the phenotypic features and DNA-DNA hybridization data, strain IR73-Li102(T) (= NBRC 106365(T) = KCTC 19783(T)) represents a novel species of the genus Actinomycetospora, for which the name Actinomycetospora iriomotensis sp. nov. is proposed.

  10. 16S rRNA-based PCR-DGGE analysis of actinomycete communities in fields with continuous cotton cropping in Xinjiang, China.

    Science.gov (United States)

    Zhang, Wei; Long, XuanQi; Huo, XiangDong; Chen, YiFeng; Lou, Kai

    2013-08-01

    The purpose of this study was to examine the variations in the microbial community structure of soil actinomycetes in fields with continuous cropping of cotton in Xinjiang Autonomous Region, China. Soil samples were collected from four depths in fields with 7-year continuous cotton cropping. The community structure of soil actinomycetes was examined using the 16S rRNA-based polymerase chain reaction-density gradient gel electrophoresis (PCR-DGGE) techniques. The microbial diversity indices of the soil samples from different depths generally decreased along with the period of continuous cotton cropping. When the period of continuous cropping of cotton reached 5 years, the diversity indices rose again and gradually stabilized at a level slightly lower than that of soils with original ecology (i.e., 0-year cotton cropping). Cluster analysis showed that at the 1-20-cm depth, the actinomycete community structure of the soil subjected to 1-year cotton cropping was similar to that of soil subjected to 0-year cotton cropping, whereas that of soils after 3-year continuous cotton cropping showed high similarity. At the 21-40-cm depth, the actinomycete community structure showed various changes but generally recovered to its original pattern after repeated fluctuations. Principal component analysis showed that at the 1-30-cm depth, the actinomycete community structure varied similarly regardless of the period of continuous cotton cropping. In contrast, there were no clear actinomycete community structure variation trends at the 31-40-cm soil depth. Homology comparison of sequences recovered from the DGGE bands showed that the obtained sequences shared similarities >88 %. Alignment with the known homologous sequences indicated a lack of microorganisms related to soil-borne cotton diseases. Continuous cotton cropping exerted significant influences on the community structure of soil actinomycetes in Xinjiang Autonomous Region, which were largely determined by the soil depth and

  11. 植物内生放线菌多样性研究进展%Research Progress in Diversity of Endophytic Actinomycetes

    Institute of Scientific and Technical Information of China (English)

    冯天祥; 陆可茵; 陆兰依塔; 陈海敏; 盛清

    2015-01-01

    植物内生放线菌作为植物内生菌资源的一大类,具有丰富的多样性,其次级代谢产物也十分丰富,而且还具有各种生物学活性。通过了解植物内生放线菌资源的多样性,可以对其产生的生物活性物质进行筛选和分析,从而筛选出具有良好药理活性的物质应用于临床。从植物内生放线菌宿主植物、组织分布、种类和数量以及活性基因4个方面综述了植物内生放线菌的多样性资源,介绍植物内生放线菌新的微生物物种,总结了植物内生放线菌目前研究中存在的问题及展望。%Endophytic actinomycetes display abundant diversity and have a capacity to produce a huge number of sec-ondary metabolites exhibiting a broad variety of biological activities as a kind of endophyte resources.Biological activi-ties produced by endophytic actinomycetes are screened and analyzed via understanding the biodiversity of endophytic actinomycetes resources so that favorable pharmacological active substances were screened for clinical diseases.Re-cently, with the development of researches about endophytic actinomycetes, many novel microorganism species were reported.This review focuses on diversity resources of endophytic actinomycetes from four aspects:host plants of en-dophytic actinomycetes, distribution in host plants tissue, species and number diversity and active gene and introduces the researches of novel endophytic actinomycetes species.Finally, this article summarizes the problems of endophytic actinomycetes at present researches and gives the possible further research direction.Endophytic actinomycetes which are emerging microbial resources have an important effect on science researches and progress of mankind.

  12. Investigation on Actinomycetes Population in Yuxi Normal University%玉溪师范学院校园内土壤放线菌资源调查

    Institute of Scientific and Technical Information of China (English)

    陈艳; 李淑英; 刘家忠; 李红梅; 陈丽君; 刘菊安

    2012-01-01

    [Objective] Actinomycetes population in Yuxi normal university was investigated to provide information on the development and the utilization of actinomycetes resource. [Method] Ten soil samples from plant rhizosphere in Yuxi normal university were collected. Actinomycetes were isolated by spreading the samples on Gao 1 agar medium. Isolaets were identified at genus level.[Result] The number of actinomycetes was different in different plant rhizosphere. Streptomyces dominated. [Conclusion] Actinomycetes populations were rich in the soil of Yuxi normal university. Streptomyces dominated, mainly for the inerogriseus and the griseofuscus.%[目的]调查玉溪师范学院校园内土壤的放线菌资源,为云南放线菌资源开发利用提供资料.[方法]从玉溪师范学院校园内采集10种植物根际土壤,采用涂布平板法分离,用高氏一号培养基培养,采用国内通用的方法进行鉴定.[结果]不同植物根际分离到的放线菌数量不同;链霉菌占绝对优势.[结论]玉溪师院校园内土壤中放线菌资源丰富,链霉菌占绝对优势,主要为烬灰类群、灰褐类群.

  13. Complete genome of the cellulolytic thermophile Acidothermus cellulolyticus 11B provides insights into its ecophysiological and evolutionary adaptations

    Energy Technology Data Exchange (ETDEWEB)

    Xie, Gary [Los Alamos National Laboratory; Detter, John C [Los Alamos National Laboratory; Bruce, David C [Los Alamos National Laboratory; Challacombe, Jean F [Los Alamos National Laboratory; Brettin, Thomas S [Los Alamos National Laboratory; Necsulea, Anamaria [UNIV LYON; Daubin, Vincent [UNIV LYON; Medigue, Claudine [GENOSCOPE; Adney, William S [NREL; Xu, Xin C [UC DAVIS; Lapidus, Alla [JGI; Pujic, Pierre [UNIV LYON; Berry, Alison M [UC DAVIS; Barabote, Ravi D [UC DAVIS; Leu, David [UC DAVIS; Normand, Phillipe [UNIV LYON

    2009-01-01

    We present here the complete 2.4 MB genome of the actinobacterial thermophile, Acidothermus cellulolyticus 11B, that surprisingly reveals thermophilic amino acid usage in only the cytosolic subproteome rather than its whole proteome. Thermophilic amino acid usage in the partial proteome implies a recent, ongoing evolution of the A. cellulolyticus genome since its divergence about 200-250 million years ago from its closest phylogenetic neighbor Frankia, a mesophilic plant symbiont. Differential amino acid usage in the predicted subproteomes of A. cellulolyticus likely reflects a stepwise evolutionary process of modern thermophiles in general. An unusual occurrence of higher G+C in the non-coding DNA than in the transcribed genome reinforces a late evolution from a higher G+C common ancestor. Comparative analyses of the A. cellulolyticus genome with those of Frankia and other closely-related actinobacteria revealed that A. cellulolyticus genes exhibit reciprocal purine preferences at the first and third codon positions, perhaps reflecting a subtle preference for the dinucleotide AG in its mRNAs, a possible adaptation to a thermophilic environment. Other interesting features in the genome of this cellulolytic, hot-springs dwelling prokaryote reveal streamlining for adaptation to its specialized ecological niche. These include a low occurrence of pseudo genes or mobile genetic elements, a flagellar gene complement previously unknown in this organism, and presence of laterally-acquired genomic islands of likely ecophysiological value. New glycoside hydrolases relevant for lignocellulosic biomass deconstruction were identified in the genome, indicating a diverse biomass-degrading enzyme repertoire several-fold greater than previously characterized, and significantly elevating the industrial value of this organism.

  14. Complete genome of the cellulolytic thermophile Acidothermus cellulolyticus 11B provides insights into its ecophysiological and evolutionary adaptations

    Energy Technology Data Exchange (ETDEWEB)

    Xie, Gary [Los Alamos National Laboratory; Detter, Chris [Los Alamos National Laboratory; Bruce, David [Los Alamos National Laboratory; Challacome, Jean F [Los Alamos National Laboratory; Brettin, Thomas S [Los Alamos National Laboratory; Barabote, Ravi D [UC DAVIS; Leu, David [UC DAVIS; Normand, Philippe [CNRS, UNIV LYON; Necsula, Anamaria [CNRS, UNIV LYON; Daubin, Vincent [CNRS, UNIV LYON; Medigue, Claudine [CNRS/GENOSCOPE; Adney, William S [NREL; Xu, Xin C [UC DAVIS; Lapidus, Alla [DOE JOINT GENOME INST.; Pujic, Pierre [CNRS, UNIV LYON; Richardson, Paul [DOE JOINT GENOME INST; Berry, Alison M [UC DAVIS

    2008-01-01

    We present here the complete 2.4 MB genome of the actinobacterial thermophile, Acidothermus cellulolyticus lIB, that surprisingly reveals thermophilic amino acid usage in only the cytosolic subproteome rather than its whole proteome. Thermophilic amino acid usage in the partial proteome implies a recent, ongoing evolution of the A. cellulolyticus genome since its divergence about 200-250 million years ago from its closest phylogenetic neighbor Frankia, a mesophilic plant symbiont. Differential amino acid usage in the predicted subproteomes of A. cellulolyticus likely reflects a stepwise evolutionary process of modern thermophiles in general. An unusual occurrence of higher G+C in the non-coding DNA than in the transcribed genome reinforces a late evolution from a higher G+C common ancestor. Comparative analyses of the A. cellulolyticus genome with those of Frankia and other closely-related actinobacteria revealed that A. cellulolyticus genes exhibit reciprocal purine preferences at the first and third codon positions, perhaps reflecting a subtle preference for the dinucleotide AG in its mRNAs, a possible adaptation to a thermophilic environment. Other interesting features in the genome of this cellulolytic, hot-springs dwelling prokaryote reveal streamlining for adaptation to its specialized ecological niche. These include a low occurrence of pseudogenes or mobile genetic elements, a flagellar gene complement previously unknown in this organism, and presence of laterally-acquired genomic islands of likely ecophysiological value. New glycoside hydrolases relevant for lignocellulosic biomass deconstruction were identified in the genome, indicating a diverse biomass-degrading enzyme repertoire several-fold greater than previously characterized, and significantly elevating the industrial value of this organism.

  15. Enzymatic activity of the cellulolytic complex produced by Trichoderma reesei. Enzymatic hydrolysis of cellulose; Actividad enzimatica del complejo celulolitico producido por Trichoderma reesei. Hidrolisis enzimatica de la celulosa

    Energy Technology Data Exchange (ETDEWEB)

    Alfonsel, M.; Negro, M. J.; Saez, R.; Martin, C.

    1986-07-01

    The enzymatic activity characterization of the cellulolytic complex obtained from Trichoderma reesei QM 9414 and the influence of the enzymatic hydrolysis conditions on the hydrolysis yield are studied. Pure cellulose and native or alkali pretreated biomass Onopordum nervosum have been used as substrates. The values of pH, temperature, substrate concentration and enzyme-substrate ratio for the optimum activity of that complex, evaluated as glucose and reducing sugars production, have been selected. Previous studies on enzymatic hydrolysis of 0. nervosum have shown a remarkable effect of the alkaline pretreatments on the final hydrolysis yield. (Author) 10 refs.

  16. Antifungal Production of a Strain of Actinomycetes spp Isolated from the Rhizosphere of Cajuput Plant: Selection and Detection of Exhibiting Activity Against Tested Fungi

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    Alimuddin A

    2015-11-01

    Full Text Available Actinomycetes are bacteria known to constitute a large part of the rhizosphere microbiota. Their isolation is an important step for screening of new bioactive compounds. Culturable actinomycetes populations from cajuput plant rhizosphere soils in Wanagama I Forest UGM Yogyakarta were collected to study about their antifungal activity. Among 17 of a total 43 isolates that showed activity were screened for producing antifungi substances. Screening for antifungal activity of isolates were performed with dual culture bioassay in vitro. One isolate that was designated as Streptomyces sp.GMR-22 was the strongest against all tested fungi and appeared promising for a sources of antifungal. Culture’s supernatant and mycelia were extracted with chloroform, ethyl acetate and methanol, respectively. Antifungal activity of crude extracts was tested by diffusion method against tested fungi. The result indicates that isolates of actinomycetes from cajuput plant rhizosphere could be an interesting sources of antifungal bioactive substances.

  17. [Study of marine actinomycetes isolated from the central coast of Peru and their antibacterial activity against methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus faecalis].

    Science.gov (United States)

    León, Jorge; Aponte, Juan José; Rojas, Rosario; Cuadra, D'Lourdes; Ayala, Nathaly; Tomás, Gloria; Guerrero, Marco

    2011-06-01

    To determine the antimicrobial potential of marine actinomycetes against drug-resistant pathogens represented by strains of methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecalis (VRE). Strains of actinomycetes (29) isolated from marine sediment were evaluated by their characteristics in two culture media and by testing their inhibitory capacity by in vitro antagonism against multi-drug resistant (MDR) pathogenic bacteria for MRSA and VRE. Organic extracts of 3 selected actinomicetes were processed to determine the minimum inhibitory concentration (MIC) of the active compound. Most isolated actinomycetes belong to a homogeneous group of write-gray actinomycetes with a good growth in Marine Agar. The inhibitory rates of the isolates were above 85% for both pathogens with inhibition zones greater than 69 and 78 mm in diameter for MRSA and VRE respectively. Dichloromethane extracts of 3 isolates (I-400A, B1-T61, M10-77) showed strong inhibitory activity of both pathogens, M10-77 being the highest actinomycete strain with antibiotic activity against methicillin-resistant S. aureus ATCC 43300 and vancomycin-resistant E. faecalis ATCC 51299 with a minimum inhibitory concentrations (MIC) of 7.9 and 31.7 μg/ml respectively. Phylogenetic analysis of M10-77 strain showed 99% similarity with the marine species Streptomyces erythrogriseus. Marine sediments of the central coast of Peru, are a source of actinomycetes strains showing high capacity to produce bioactive compounds able to inhibit pathogens classified as multi-drug-resistant such as methicillin-resistant S. aureus and vancomycin-resistant E. faecalis.

  18. Nonomuraea flavida sp. nov., a novel species of soil actinomycete isolated from Aconitum napellus rhizosphere.

    Science.gov (United States)

    Chen, Shaofeng; Shi, Jindi; Li, Dan; Wu, Yingying; Huang, Yaojian

    2015-11-01

    A novel actinomycete strain, YN-5-1T, isolated from the rhizosphere soil of a medicinal plant, Aconitum napellus, was characterized by a polyphasic approach to determine its taxonomic position. The strain showed highest 16S rRNA gene sequence similarities of 97.3, 97.2 and 97.1 % to Nonomuraea turkmeniaca DSM 43926T, Nonomuraea ferruginea DSM 43553T and Nonomuraea candida DSM 45086T, respectively. A wide range of genotypic and phenotypic characteristics, as well as levels of DNA-DNA relatedness between strain YN-5-1T and N. turkmeniaca DSM 43926T (57.46 %), N. ferruginea DSM 43553T (53.50 %) and N. candida DSM 45086T (48.80 %), distinguished the novel isolate from its closest phylogenetic neighbours. The morphological characteristics of strain YN-5-1T were typical of the genus Nonomuraea. Chemotaxonomic characteristics, such as diagnostic diamino acid of the peptidoglycan, whole-cell sugars, phospholipid type, major menaquinone and major fatty acids, further supported the assignment of strain YN-5-1T to the genus Nonomuraea. The G+C content of the genomic DNA was 72.1 mol%. Based on the above data, strain YN-5-1T is considered to represent a novel species of the genus Nonomuraea, for which the name Nonomuraea flavida sp. nov. is proposed. The type strain is YN-5-1T ( = CCTCC AB 2012909T = KCTC 29143T).

  19. Actinopolyspora biskrensis sp. nov., a novel halophilic actinomycete isolated from Northern Sahara.

    Science.gov (United States)

    Saker, Rafika; Bouras, Noureddine; Meklat, Atika; Zitouni, Abdelghani; Schumann, Peter; Spröer, Cathrin; Klenk, Hans-Peter; Sabaou, Nasserdine

    2015-03-01

    A novel halophilic, filamentous actinomycete, designated H254(T), was isolated from a Saharan soil sample collected from Biskra (Northern Sahara), and subjected to a polyphasic taxonomic characterization. The strain is Gram-positive, aerobic, and halophilic, and the optimum NaCl concentration for growth is 15-20 % (w/v). The cell-wall hydrolysate contained meso-diaminopimelic acid, and the diagnostic whole-cell sugars were arabinose and galactose. The diagnostic phospholipid detected was phosphatidylcholine, and MK-9(H4) was the predominant menaquinone. The major fatty acid profiles were anteiso-C17:0 (32.8 %), C15:0 (28 %), and iso-C17:0 (12.3 %). Comparative analysis of the 16S rRNA gene sequences revealed that the strain H254(T) formed a well-separated sub-branch within the radiation of the genus Actinopolyspora, and the microorganism was most closely related to Actinopolyspora saharensis DSM 45459(T) (99.2 %), Actinopolyspora halophila DSM 43834(T) (99.1 %), and Actinopolyspora algeriensis DSM 45476(T) (99.0 %). Nevertheless, the strain had relatively lower mean values for DNA-DNA relatedness with the above strains (57.2, 68.4, and 45.6 %, respectively). Based on phenotypic features and phylogenetic position, we propose that strain H254(T) represents a novel species of the genus Actinopolyspora, for which the name Actinopolyspora biskrensis sp. nov. is proposed. The type strain of A. biskrensis is strain H254(T) (=DSM 46684(T) =CECT 8576(T)).

  20. Diketopiperazine Derivatives from the Marine-Derived Actinomycete Streptomyces sp. FXJ7.328

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    Weiming Zhu

    2013-03-01

    Full Text Available Five new diketopiperazine derivatives, (3Z,6E-1-N-methyl-3-benzy lidene-6-(2S-methyl-3-hydroxypropylidenepiperazine-2,5-dione (1, (3Z,6E-1-N-methyl-3-benzylidene-6-(2R-methyl-3-hydroxypropylidenepiperazine-2,5-dione (2, (3Z,6Z-3- (4-hydroxybenzylidene-6-isobutylidenepiperazine-2,5-dione (3, (3Z,6Z-3-((1H-imidazol-5-yl-methylene-6-isobutylidenepiperazine-2,5-dione (4, and (3Z,6S-3-benzylidene-6-(2S-but-2-ylpiperazine-2,5-dione (5, were isolated from the marine-derived actinomycete Streptomyces sp. FXJ7.328. The structures of 1–5 were determined by spectroscopic analysis, CD exciton chirality, the modified Mosher’s, Marfey’s and the C3 Marfey’s methods. Compound 3 showed modest antivirus activity against influenza A (H1N1 virus with an IC50 value of 41.5 ± 4.5 μM. In addition, compound 6 and 7 displayed potent anti-H1N1 activity with IC50 value of 28.9 ± 2.2 and 6.8 ± 1.5 μM, respectively. Due to the lack of corresponding data in the literature, the 13C NMR data of (3Z,6S-3-benzylidene-6-isobutylpiperazine-2,5-dione (6 were also reported here for the first time.

  1. Thermodynamics of a Ca(2+)-dependent highly thermostable alkaline protease from a haloalkliphilic actinomycete.

    Science.gov (United States)

    Gohel, S D; Singh, S P

    2015-01-01

    An alkaline protease from salt-tolerant alkaliphilic actinomycetes, Nocardiopsis alba OK-5 was purified by a single-step hydrophobic interaction chromatography and characterized. The purified protease with an estimated molecular mass of 20 kDa was optimally active at 70 °C in 0-3 M NaCl and 0-100 mM Ca(2+) displaying significant stability at 50-80 °C. The enzyme was stable at 80 °C in 100 mM Ca(2+) with Kd of 17 × 10(-3) and t1/2 of 32 min. The activation energy (Ea), enthalpy (ΔH*), and entropy (ΔS*) for the protease deactivation calculated in the presence of 200 mM Ca(2+) were 38.15 kJ/mol, 35.49 kJ/mol and 183.48 J/mol, respectively. The change in free energy (ΔG*) for protease deactivation at 60 °C in 200 mM Ca(2+) was 95.88 kJ/mol. Decrease in ΔH* reflected reduced cooperativity of deactivation and unfolding. The enzyme was intrinsically stable that counteracted heat denaturation by a weak cooperativity during the unfolding. Further, the enzyme was highly stable in the presence of various cations, surfactants, H2O2, β-mercaptoethanol, and commercial detergents. The compatibility of the enzyme with various cations, surfactants, and detergent matrices suggests its suitability as an additive in the detergents and peptide synthesis.

  2. Saccharothrix lopnurensis sp. nov., a filamentous actinomycete isolated from sediment of Lop Nur.

    Science.gov (United States)

    Li, Yu-Qian; Liu, Lan; Cheng, Cong; Shi, Xiao-Han; Lu, Chun-Yan; Dong, Zhou-Yan; Salam, Nimaichand; An, Deng-Di; Li, Wen-Jun

    2015-10-01

    A novel actinomycete strain, designated YIM LPA2h(T), was isolated from a sediment sample collected from Lop Nur, Xinjiang Uygur Autonomous Region, North-West China. The taxonomic position of strain YIM LPA2h(T) was investigated by a polyphasic approach. The morphological and chemotaxonomic properties of the isolate were in accordance with the members of the genus Saccharothrix. The 16S rRNA gene sequence of the strain showed highest similarities to Saccharothrix yanglingensis (98.6 %), Saccharothrix longispora (98.4 %) and Saccharothrix hoggarensis (98.3 %). However, the DNA-DNA hybridization values between the new isolate YIM LPA2h(T) and S. yanglingensis, S. longispora and S. hoggarensis were significantly below 70 %. Strain YIM LPA2h(T) was found to contain meso-diaminopimelic acid as diagnostic diamino acid. The major sugars in whole-cell hydrolysates were rhamnose, galactose, mannose, glucose and fructose. The major polar lipids were identified as phosphatidylethanolamine, phosphatidylhydroxylethanolamine, diphosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannoside. The predominant respiratory menaquinones were MK-9 (H4) and MK-10 (H4). The major fatty acids were C17:1 ω8c (15 %), iso-C15:0 (12 %), anteiso-C15:0 (12 %) and summed feature 3 (C16:1 ω6c/C16:1 ω7c, 10 %). The genomic DNA G+C content of strain YIM LPA2h(T) was determined to be 75 mol %. The genotypic and phenotypic results suggest that strain YIM LPA2h(T) represents a novel species of the genus Saccharothrix, for which the name Saccharothrix lopnurensis sp. nov. is proposed. The type strain is YIM LPA2h(T) (=CGMCC 4.7246(T)=KCTC 39545(T)).

  3. Identification and Antibiosis of a Novel Actinomycete Strain RAF-11 Isolated From Iraqi Soil.

    Directory of Open Access Journals (Sweden)

    Rabah Forar Laidi

    2013-04-01

    Full Text Available A total of 35 actinomycetes strains were isolated from and around Baghdad, Iraq, at a depth of 5-10 m, by serial dilution agar plating method. Nineteen out of them showed noticeable antimicrobial activities against at least, to one of the target pathogens. Five among the nineteen were active against both Gram positive and Gram negative bacteria, yeasts and moulds. The most active isolate, strain RAF-11, based on its largest zone of inhibition and strong antifungal activity, especially against Candida albicans and Aspergillus niger, the causative of candidiasis and aspergillosis respectively, was selected for identification. Morphological and chemical studies indicated that this isolate belongs to the genus Streptomyces. Analysis of the 16S rDNA sequence showed a high similarity, 98 %, with the most closely related species, Streptomyces labedae NBRC 15864T/AB184704, S. erythrogriseus LMG 19406T/AJ781328, S. griseoincarnatus LMG 19316T/AJ781321 and S. variabilis NBRC 12825T/AB184884, having the closest match. From the taxonomic features, strain RAF-11 matched with S. labedae, in the morphological, physiological and biochemical characters, however it showed significant differences in morphological characteristics with this nearest species, S. labedae, which encourage us to consider our starin as a novel isolate and was given the suggested name, Streptomyces labedae strain RAF-11. ISP-4 broth medium supplemented with glucose and soybean powder at concentrations of 1g % and 0.1g % as carbon and nitrogen sources respectively, for 120h incubation at 28 °C, increased the active compounds production, where we recorded a strong activity against yeasts, 42mm inhibition zone against Candida albicans, 41mm against C. pseudotropicalis, 40mm against C. tropicalis, followed by 38mm against Rhodotorula minota and Aspergillus niger then, 35mm against both Aspergillus flavus and Bacillus subtilis. N-butanol was best solvent for antibiotic extraction compared to

  4. Saccharopolyspora griseoalba sp. nov., a novel actinomycete isolated from the Dead Sea.

    Science.gov (United States)

    Jiang, Yingying; Wei, Xiaomin; Chen, Xiu; Jiang, Yi; Xue, Quanhong; Lai, Hangxian; Jiang, Chenglin

    2016-12-01

    A novel halotolerant actinomycete, designated strain AFM 10238(T), was isolated from a sediment sample collected from the Dead Sea of Israel. The isolate grew at 15-45 °C, pH 6-12 and with 0-15 % (w/v) NaCl. Strain AFM 10238(T) contains meso-diaminopimelic acid as cell wall diamino acid, and galactose and arabinose as the whole cell sugars. The major polar lipids are phosphatidylcholine, phosphatidylglycerol, and diphosphatidylglycerol. Major fatty acids are iso-C16:0, iso-C17:0, iso-C15:0, anteiso-C17:0 and C17:1 ω8c. MK-9(H4) is the predominant menaquinone and the DNA G + C content is 72.7 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain AFM10238(T) belongs to the genus Saccharopolyspora. The 16S rRNA gene sequence similarity between strain AFM 10238(T) and its close neighbours, Saccharopolyspora halophila YIM 90500(T) , Saccharopolyspora spinosa DSM 44228(T), Saccharopolyspora dendranthemae KLBMP 1305(T) and Saccharopolyspora cebuensis DSM 45019(T) were 98.2, 97.2, 97.1 and 97.0 %, respectively. Sequence similarities to other type strains of this genus were below 97 %. DNA-DNA relatedness data, together with phenotypic and chemotaxonomic differences, clearly distinguished the isolate from its close neighbours. On the basis of the data from this polyphasic analysis, a novel species Saccharopolyspora griseoalba sp. nov. is proposed. The type strain is AFM 10238(T) (= DSM 46,663 = CGMCC 4.7124).

  5. Amycolatopsis flava sp. nov., a halophilic actinomycete isolated from Dead Sea.

    Science.gov (United States)

    Wei, Xiaomin; Jiang, Yingying; Chen, Xiu; Jiang, Yi; Lai, Hangxian

    2015-10-01

    A novel halophilic, filamentous actinomycete, designated strain AFM 10111(T), was isolated from a sediment sample collected from the Dead Sea of Israel and its taxonomic position was established by using a polyphasic taxonomic approach. The isolate grew at 20-35 °C, pH 5-12 and with 1-30 % NaCl. The substrate mycelium is white or yellow, well developed, branched and fragments into squarish, rod-like elements. The isolate contained meso-diaminopimelic acid as cell-wall diamino acid, and arabinose and galactose as whole-cell sugars. The major menaquinone was MK-9(H4). The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine phosphatidylmethylethanolamine and one unidentified phospholipid. Major fatty acids were iso-C16:0, iso-C16:1 H, C17:1 ω6c. The DNA G + C content was 67.7 mol %. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain AFM 10111(T) belongs to the genus Amycolatopsis, and formed a distinct clade with Amycolatopsis marina CGMCC 4.3568(T) and Amycolatopsis palatopharyngis CGMCC 4.1729(T), with the sequence similarity 98.4 and 98.6 %. The level of DNA-DNA relatedness between the strain AFM 10111(T) and A. marina CGMCC 4.3568(T) and A. palatopharyngis CGMCC 4.1729(T) were 46.9 ± 3.08 and 49.4 ± 1.25 %. The combined genotypic and phenotypic data indicate that strain AFM 10111(T) represents a novel species of the genus Amycolatopsis, for which the name Amycolatopsis flava sp. nov. is proposed. The type strain is AFM 10111(T) (= DSM 46658(T) = CGMCC 4.7123(T)).

  6. Discovery of phosphonic acid natural products by mining the genomes of 10,000 actinomycetes.

    Science.gov (United States)

    Ju, Kou-San; Gao, Jiangtao; Doroghazi, James R; Wang, Kwo-Kwang A; Thibodeaux, Christopher J; Li, Steven; Metzger, Emily; Fudala, John; Su, Joleen; Zhang, Jun Kai; Lee, Jaeheon; Cioni, Joel P; Evans, Bradley S; Hirota, Ryuichi; Labeda, David P; van der Donk, Wilfred A; Metcalf, William W

    2015-09-29

    Although natural products have been a particularly rich source of human medicines, activity-based screening results in a very high rate of rediscovery of known molecules. Based on the large number of natural product biosynthetic genes in microbial genomes, many have proposed "genome mining" as an alternative approach for discovery efforts; however, this idea has yet to be performed experimentally on a large scale. Here, we demonstrate the feasibility of large-scale, high-throughput genome mining by screening a collection of over 10,000 actinomycetes for the genetic potential to make phosphonic acids, a class of natural products with diverse and useful bioactivities. Genome sequencing identified a diverse collection of phosphonate biosynthetic gene clusters within 278 strains. These clusters were classified into 64 distinct groups, of which 55 are likely to direct the synthesis of unknown compounds. Characterization of strains within five of these groups resulted in the discovery of a new archetypical pathway for phosphonate biosynthesis, the first (to our knowledge) dedicated pathway for H-phosphinates, and 11 previously undescribed phosphonic acid natural products. Among these compounds are argolaphos, a broad-spectrum antibacterial phosphonopeptide composed of aminomethylphosphonate in peptide linkage to a rare amino acid N(5)-hydroxyarginine; valinophos, an N-acetyl l-Val ester of 2,3-dihydroxypropylphosphonate; and phosphonocystoximate, an unusual thiohydroximate-containing molecule representing a new chemotype of sulfur-containing phosphonate natural products. Analysis of the genome sequences from the remaining strains suggests that the majority of the phosphonate biosynthetic repertoire of Actinobacteria has been captured at the gene level. This dereplicated strain collection now provides a reservoir of numerous, as yet undiscovered, phosphonate natural products.

  7. Rare actinomycetes Nocardia caishijiensis and Pseudonocardia carboxydivorans as endophytes, their bioactivity and metabolites evaluation.

    Science.gov (United States)

    Tanvir, Rabia; Sajid, Imran; Hasnain, Shahida; Kulik, Andreas; Grond, Stephanie

    2016-04-01

    Two strains identified as Nocardia caishijiensis (SORS 64b) and Pseudonocardia carboxydivorans (AGLS 2) were isolated as endophytes from Sonchus oleraceus and Ageratum conyzoides respectively. The analysis of their extracts revealed them to be strongly bioactive. The N. caishijiensis extract gave an LC50 of 570 μg/ml(-1) in the brine shrimp cytotoxicity assay and an EC50 of 0.552 μg/ml(-1) in the DPPH antioxidant assay. Antimicrobial activity was observed against Methicillin resistant Staphlococcus aureus (MRSA) and Escherichia coli ATCC 25922 (14 mm), Klebsiella pneumoniae ATCC 706003 (13 mm), S. aureus ATCC 25923 (11 mm) and Candida tropicalis (20 mm). For the extract of P. carboxydivorans the EC50 was 0.670 μg/ml(-1) and it was observed to be more bioactive against Bacillus subtilis DSM 10 ATCC 6051 (21 mm), C. tropicalis (20 mm), S. aureus ATCC 25923 (17 mm), MRSA (17 mm), E. coli K12 (W1130) (16 mm) and Chlorella vulgaris (10 mm). The genotoxicity testing revealed a 20 mm zone of inhibition against the polA mutant strain E. coli K-12 AB 3027 suggesting damage to the DNA and polA genes. The TLC and bioautography screening revealed a diversity of active bands of medium polar and nonpolar compounds. Metabolite analysis by HPLC-DAD via UV/vis spectral screening suggested the possibility of stenothricin and bagremycin A in the mycelium extract of N. caishijiensis respectively. In the broth and mycelium extract of P. carboxydivorans borrelidin was suggested along with α-pyrone. The HPLC-MS revealed bioactive long chained amide derivatives such as 7-Octadecenamide, 9, 12 octadecandienamide. This study reports the rare actinomycetes N. caishijiensis and P. carboxydivorans as endophytes and evaluates their bioactive metabolites.

  8. Sphaerisporangium dianthi sp. nov., an endophytic actinomycete isolated from a root of Dianthus chinensis L.

    Science.gov (United States)

    Xing, Jia; Liu, Chongxi; Zhang, Yuejing; He, Hairong; Zhou, Ying; Li, Lianjie; Zhao, Junwei; Liu, Shuanghe; Wang, Xiangjing; Xiang, Wensheng

    2015-01-01

    A novel actinomycete, designated strain NEAU-CY18(T), was isolated from the root of a Chinese medicinal plant Dianthus chinensis L and subjected to a polyphasic taxonomic study. The novel strain was found to develop spherical sporangia with non-motile spores on aerial mycelium. The cell-wall peptidoglycan was found to contain meso-diaminopimelic acid. The whole-cell sugars were identified as madurose, mannose, ribose, galactose and glucose. The phospholipid profile was found to contain diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylethanolamine, hydroxy-phosphatidylmethylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannosides and an unidentified phospholipid. The predominant menaquinones were identified as MK-9(H4), MK-9(H2) and MK-9(H6). The major fatty acids were identified as C17:0 10-methyl, iso-C16:0 and C16:0. EzTaxon-e analysis of the 16S rRNA gene sequence indicated that the strain belongs to the genus Sphaerisporangium and was most closely related to Sphaerisporangium cinnabarinum JCM 3291(T) (98.9 %) and Sphaerisporangium melleum JCM 13064(T) (98.3 %). Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain NEAU-CY18(T) forms a monophyletic clade with S. cinnabarinum JCM 3291(T), an association that was supported by a bootstrap value of 97 % in the neighbour-joining tree and also recovered with the maximum-likelihood algorithm. Comparisons of some phenotypic properties and low DNA-DNA relatedness values enabled the strain to be differentiated from S. cinnabarinum JCM 3291(T) and S. melleum JCM 13064(T). Therefore, it is concluded that strain NEAU-CY18(T) represents a novel Sphaerisporangium species, for which the name Sphaerisporangium dianthi sp. nov. is proposed. The type strain is NEAU-CY18(T) ( = CGMCC 4.7132(T) = DSM 46736(T)).

  9. Streptomyces gilvifuscus sp. nov., an actinomycete that produces antibacterial compounds isolated from soil.

    Science.gov (United States)

    Nguyen, uan Manh; Kim, Jaisoo

    2015-10-01

    This study describes a novel actinomycete, designated T113T, which was isolated from forest soil in Pyeongchang-gun, Republic of Korea, and is an aerobic, Gram-stain-positive actinobacterium that forms flexibilis chains of smooth, elliptical or short rod-shaped spores. The results of 16S rRNA sequence analysis indicated that strain T113T exhibited high levels of similarity to previously characterized species of the genus Streptomyces (98.19–98.89 %, respectively). However, the results of phylogenetic and DNA–DNA hybridization analyses confirmed that the organism represented a novel member of the genus Streptomyces. Furthermore, using chemotaxonomic and phenotypic analyses it was demonstrated that the strain exhibited characteristics similar to those of other members of the genus Streptomyces. The primary cellular fatty acids expressed by this strain included anteiso-C15 : 0, anteiso-C17 : 0, iso-C15 : 0 and iso-C16 : 0. While diphosphatidylglycerol and phosphatidylethanolamine were the predominant lipids expressed by strain T113T, moderate amounts of phosphatidylinositol and phosphatidylinositol mannoside were also detected. Whole-cell hydrolysates contained glucose and ribose, and the predominant menaquinone detected was MK-9 (H6); however, moderate amounts of MK-9 (H8) and trace amounts of MK-10 (H2) and MK-10 (H4) were also detected. We therefore propose that strain T113T be considered as representing a novel species of the genus Streptomyces and propose the name Streptomyces gilvifuscus sp. nov. for this species, with strain T113T ( = KEMB 9005-213T = KACC 18248T = NBRC 110904T) being the type strain.

  10. Streptomyces maoxianensis sp. nov., a novel actinomycete isolated from soil in Maoxian, China.

    Science.gov (United States)

    Guan, Xuejiao; Liu, Chongxi; Zhao, Junwei; Fang, Baozhu; Zhang, Yuejing; Li, Lianjie; Jin, Pinjiao; Wang, Xiangjing; Xiang, Wensheng

    2015-05-01

    A novel actinomycete, designated strain NEAU-Spg16(T), was isolated from a soil sample from a pine forest in Songpinggou, Maoxian, southwest China. A polyphasic taxonomic study was carried out to establish the status of strain NEAU-Spg16(T). The cell wall peptidoglycan was found to contain LL-diaminopimelic acid and glycine. The major menaquinones were identified as MK-9(H8), MK-9(H4) and MK-9(H6). The phospholipid profile was found to contain diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and an unidentified phospholipid. The major fatty acids were identified as iso-C(16:0), C(18:0), C(16:0) and iso-C(15:0). 16S rRNA gene sequence similarity studies showed that strain NEAU-Spg16(T) belongs to the genus Streptomyces with the highest sequence similarity to Streptomyces nitrosporeus DSM 40023(T) (98.6%). However, phylogenetic analysis based on the 16S rRNA gene sequence indicated that it is most closely related to Streptomyces scopuliridis DSM 41917(T) (98.2% sequence similarity). A combination of DNA-DNA hybridization results and some phenotypic characteristics indicated that strain NEAU-Spg16(T) can be clearly differentiated from S. scopuliridis DSM 41917(T) and S. nitrosporeus DSM 40023(T). Therefore, it is concluded that strain NEAU-Spg16(T) represents a novel species of the genus of Streptomyces, for which the name Streptomyces maoxianensis sp. nov. is proposed. The type stain is NEAU-Spg16(T) (=CGMCC 4.7139(T) = DSM 42137(T)).

  11. Streptomyces formicae sp. nov., a novel actinomycete isolated from the head of Camponotus japonicus Mayr.

    Science.gov (United States)

    Bai, Lu; Liu, Chongxi; Guo, Lifeng; Piao, Chenyu; Li, Zhilei; Li, Jiansong; Jia, Feiyu; Wang, Xiangjing; Xiang, Wensheng

    2016-02-01

    During a screening for novel and biotechnologically useful actinobacteria in insects, a novel actinomycete with antifungal activity, designated strain 1H-GS9(T), was isolated from the head of a Camponotus japonicus Mayr ant, which were collected from Northeast Agricultural University (Harbin, Heilongjiang, China). Strain 1H-GS9(T) was characterised using a polyphasic approach. The organism was found to have morphological and chemotaxonomic characteristics typical of members of the genus Streptomyces. 16S rRNA gene sequence similarity studies showed that strain 1H-GS9(T) belongs to the genus Streptomyces with high sequence similarities to Streptomyces scopuliridis DSM 41917(T) (98.8 %) and Streptomyces mauvecolor JCM 5002(T) (98.6 %). However, phylogenetic analysis based on the 16S rRNA gene sequence indicated that it forms a monophyletic clade with Streptomyces kurssanovii JCM 4388(T) (98.6 %), Streptomyces xantholiticus JCM 4282(T) (98.6 %) and Streptomyces peucetius JCM 9920(T) (98.5 %). Thus, a combination of DNA-DNA hybridization experiments and phenotypic tests were carried out between strain 1H-GS9(T) and the above-mentioned five strains, which further clarified their relatedness and demonstrated that strain 1H-GS9(T) could be distinguished from these strains. Therefore, the strain is concluded to represent a novel species of the genus Streptomyces, for which the name Streptomyces formicae sp. nov. is proposed. The type strain is 1H-GS9(T) (=CGMCC 4.7277(T) = DSM 100524(T)).

  12. Streptomyces bohaiensis sp. nov., a novel actinomycete isolated from Scomberomorus niphonius in the Bohai Sea.

    Science.gov (United States)

    Pan, Hua-Qi; Cheng, Juan; Zhang, Dao-Feng; Yu, Su-Ya; Khieu, Thi-Nhan; Son, Chu Ky; Jiang, Zhao; Hu, Jiang-Chun; Li, Wen-Jun

    2015-04-01

    A novel actinomycete strain, designated 11A07(T), was isolated from young Scomberomorus niphonius in the Bohai Sea. Basic local alignment search tool analyses showed that this isolate had the highest 16S rRNA gene sequence similarity of 97.41% with Streptomyces rimosus subsp. paromomycinus DSM 41429(T). Phylogenetic tree revealed that strain 11A07(T) formed a distinct lineage clustered with Streptomyces panacagri Gsoil 519(T), Streptomyces sodiiphilus YIM 80305(T) and Streptomyces albus subsp. albus NRRL B-2365(T) having similarities of 97.30%, 97.10% and 96.83%, respectively. Multilocus sequence analysis further demonstrated that the new isolate was different from the selected representatives of Streptomyces as a separate phylogenetic line. Strain 11A07(T) produced straight or rectiflexibile spore chains with smooth surface, white aerial mycelia and brown diffusible pigments on international streptomyces project 2 medium. Maximum tolerated NaCl concentration for growth was 11.0%. Whole-cell sugars were mannose, ribose, glucose, galactose and xylose. The predominant menaquinones were MK-9(H2), MK-9(H4) and MK-9 (H6). The fatty-acid profile contained iso-C16:0, C18:0 10-methyl (tuberculostearic acid) and anteiso-C17:0 as the major compositions. The polar lipids consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannoside and an unknown phospholipid. The G+C content of the genomic DNA was 71.4 mol%. These morphological, phenotypic and chemotaxonomic properties showed that strain 11A07(T) could be readily distinguished from the most closely related members of the genus Streptomyces. Thus, based on the polyphasic taxonomic data, strain 11A07(T) (=JCM 19630(T)=CCTCC AA 2013020(T)=KCTC 29263(T)) represents a novel species within the genus Streptomyces, for which the name Streptomyces bohaiensis sp. nov. is proposed.

  13. Streptomyces bryophytorum sp. nov., an endophytic actinomycete isolated from moss (Bryophyta).

    Science.gov (United States)

    Li, Chuang; Jin, Pinjiao; Liu, Chongxi; Ma, Zhaoxu; Zhao, Junwei; Li, Jiansong; Wang, Xiangjing; Xiang, Wensheng

    2016-09-01

    A novel endophytic actinomycete, designated strain NEAU-HZ10(T) was isolated from moss and characterised using a polyphasic approach. The strain was found to have morphological and chemotaxonomic characteristics typical of the genus Streptomyces. Strain NEAU-HZ10(T) formed grayish aerial mycelia, which differentiated into straight to flexuous chains of cylindrical spores. The cell wall peptidoglycan was found to contain LL-diaminopimelic acid. Predominant menaquinones were identified as MK-9(H6) and MK-9(H8). The polar lipid profile was found to consist of phosphatidylethanolamine, phosphatidylinositol and two unidentified phospholipids. The major fatty acids were identified as iso-C16:0, anteiso-C15:0 and C16:0. 16S rRNA gene sequence similarity studies showed that strain NEAU-HZ10(T) belongs to the genus Streptomyces and exhibits high sequence similarity to Streptomyces cocklensis DSM 42063(T) (98.9 %). Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain NEAU-HZ10(T) clustered with S. cocklensis DSM 42063(T), Streptomyces yeochonensis CGMCC 4.1882(T) (98.7 %), Streptomyces paucisporeus CGMCC 4.2025(T) (98.4 %) and Streptomyces yanglinensis CGMCC 4.2023(T) (98.1 %). However, a combination of DNA-DNA hybridisation results and some phenotypic characteristics indicated that strain NEAU-HZ10(T) can be distinguished from its phylogenetically closely related strains. Therefore, it is proposed that strain NEAU-HZ10(T) represents a novel species of the genus Streptomyces for which the name Streptomyces bryophytorum sp. nov. is proposed. The type strain is NEAU-HZ10(T) (= CGMCC 4.7151(T) = DSM 42138(T)).

  14. Use of dyes in solid medium for screening ligninolytic activity of selective actinomycetes

    Energy Technology Data Exchange (ETDEWEB)

    Chahal, D.S.; Kluepfel, D.; Morosoli, R. [Universite du Quebec (Canada)] [and others

    1995-12-31

    Lignin, a three-dimensional biopolymer, not only encrusts the cellulose microfibrils in a sheath-like manner, but is also bonded physically and chemically to the plant polysaccharides. Unless the lignin is depolymerized, solubilized, or removed, the cellulose and hemicelluloses cannot be easily hydrolyzed by respective enzymes for their bioconversion into biofuels and chemicals. By now it has been established that lignin peroxidase (LiP) of white-rot fungus Phanerochaete chrysosporium is responsible for degradation of lignin. It has been reported that LiP is produced during secondary metabolism under carbon or nitrogen limitation by this organism. In literature, usually low yields (per unit volume) of LiP with P. chrysosporium have been reported. The reasons for low yields may be attributed to insufficient nitrogen in production media, which ultimately affects the synthesis of LiP protein. Therefore, it necessitated a search for an organism that can produce a ligninolytic enzyme system during its primary metabolism, without any effect of nitrogen limitation in the fermentation medium and without supply of extra oxygen to the cultures. Glenn and Gold were the first to report that decolorization of polymeric dyes in liquid cultures is related to the lignin degradation system. They demonstrated that like lignin degradation, the decolorization of polymeric dyes by the white-rot basidiomycete P. chrysosporium occurred during secondary metabolism, was suppressed in cultures grown in the presence of high levels of nitrogen, and was strongly dependent on the oxygen concentration in the cultures. The present study was undertaken to establish if certain dyes in solid media could be used to screen ligninolytic activity of selective actinomycetes during their primary metabolism without the limitation of nitrogen in the medium.

  15. Domain Characterization of Cyclosporin Regio-Specific Hydroxylases in Rare Actinomycetes.

    Science.gov (United States)

    Woo, Min-Woo; Lee, Bo-Ram; Nah, Hee-Ju; Choi, Si-Sun; Li, Shengying; Kim, Eung-Soo

    2015-10-01

    Cytochrome P450 hydroxylase (CYP) in actinomycetes plays an important role in the biosynthesis and bioconversion of various secondary metabolites. Two unique CYPs named CYP-sb21 and CYP-pa1, which were identified from Sebekia benihana and Pseudonocardia autotrophica, respectively, were proven to transfer a hydroxyl group at the 4(th) or 9(th) N-methyl leucine position of immunosuppressive agent cyclosporin A (CsA). Interestingly, these two homologous CYPs showed different CsA regio-selectivities. CYP-sb21 exhibited preferential hydroxylation activity at the 4(th) position over the 9(th) position, whereas CYP-pa1 showed the opposite preference. To narrow down the CYP domain critical for CsA regio-selectivity, each CYP was divided into four domains, and each domain was swapped with its counterpart from the other CYP. A total of 18 hybrid CYPs were then individually tested for CsA regioselectivity. Although most of the hybrid CYPs failed to exhibit a significant change in regioselectivity in the context of CsA hydroxylation, hybrid CYP-pa1 swapped with the second domain of CYP-sb21 showed a higher preference for the 9th position. Moreover, hybrid CYPsb21 containing seven amino acids from the 2nd domain of CYP-pa1 showed higher preference for the 4(th) position. These results imply that the 2nd domain of CsA-specific CYP plays a critical role in CsA regio-selectivity, thereby setting the stage for biotechnological application of CsA regio-selective hydroxylation.

  16. Asanoa endophytica sp. nov., an endophytic actinomycete isolated from the rhizome of Boesenbergia rotunda.

    Science.gov (United States)

    Niemhom, Nantawan; Chutrakul, Chanikul; Suriyachadkun, Chanwit; Thawai, Chitti

    2016-01-12

    A novel Gram-stain-positive, non motile endophytic actinomycete, designated strain BR3-1T, which produced spore chains borne on the tip of short sporophores was isolated from the rhizome of Boesenbergia rotunda collected from Udon Thani province, Thailand. This strain was investigated for its taxonomic position using polyphasic approach. The strain contained 3-hydroxy-diaminopimelic acid and meso-diaminopimelic acid in the cell-wall peptidoglycan. The whole-cell sugars contained glucose, mannose, rhamnose, ribose and xylose. Phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannosides were found as the characteristic phospholipids. The predominant menaquinones were MK-10(H8) and MK-10(H6). The major cellular fatty acids were iso-C16:0, iso-C15:0 and anteiso-C15:0. The G+C content of the genomic DNA was 71.4 mol%. On the basis of 16S rRNA gene sequences analysis revealed that strain BR3-1T belonged to the genus Asanoa and was most closely related to Asanoa ishikariensis (99.39%), Asanoa iriomotensis (99.31%), Asanoa siamensis (99.17%), Asanoa ferruginea (98.84%) and Asanoa hainanensis (98.71%). The DNA-DNA relatedness value between the strain BR3-1T and its phylogenetically closest relatives were in the range of 15.4% ± 1.2 to 45.8% ± 2.6. In addition, some physiological and biochemical properties indicated that strain BR3-1T could be readily distinguished from all type strains in the genus Asanoa. Thus, strain BR3-1T should be represented as a novel species, for which the name Asanoa endophytica sp. nov. is proposed. The type strain is BR3-1T (=BCC 66355T =NBRC 110002T).

  17. Nonomuraea stahlianthi sp. nov., an endophytic actinomycete isolated from the stem of Stahlianthus campanulatus.

    Science.gov (United States)

    Niemhom, Nantawan; Chutrakul, Chanikul; Suriyachadkun, Chanwit; Thawai, Chitti

    2017-08-01

    A novel endophytic actinomycete, designated strain SC1-1T, was isolated from sterilized stem tissue from Stahlianthus campanulatus collected in Udon Thani province, Thailand. The isolate formed short chains of spores on aerial mycelium and presented meso-diaminopimelic acid in the cell wall peptidoglycan. Glucose, madurose, mannose, rhamnose and ribose were observed as sugars in the cells. The cell membrane phospholipids were phosphatidylethanolamine, phosphatidylmethylethanolamine, hydroxy-phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside and ninhydrin-positive phosphoglycolipids. The major menaquinones were MK-9(H4) and MK-9(H2). The main cellular fatty acids were iso-C16:0, 10-methyl C17 : 0 and C17 : 1ω6c. A high G+C content (70.7 mol%) was present in the genomic DNA. The taxonomic position based on the 16S rRNA gene sequence analysis revealed that strain SC1-1T belonged to the genus Nonomuraea and shared the highest 16S rRNA gene sequence similarity value with Nonomuraea dietziae DSM 44320T (98.82 %), followed by Nonomuraea africana IFO 14745T (98.58 %), Nonomuraea jabiensis A4036T (98.43 %), Nonomuraea endophytica YIM 65601T (98.36 %), Nonomuraea purpurea 1SM4-01T (98.34 %), Nonomuraea angiospora IFO 13155T (98.29 %), Nonomuraea roseola IFO 14685T (98.23 %) and Nonomuraea recticatena IFO 14525T (98.23 %). On the basis of the DNA-DNA hybridization relatedness and including the physiological and biochemical characteristics, strain SC1-1T should be judged as a novel species of the genus Nonomuraea, for which the name Nonomuraea stahlianthi sp. nov. is proposed. The type strain is strain SC1-1T (=BCC 66361T=NBRC 110006T).

  18. Tartrolon D, a cytotoxic macrodiolide from the marine-derived actinomycete Streptomyces sp. MDG-04-17-069.

    Science.gov (United States)

    Pérez, Marta; Crespo, Cristina; Schleissner, Carmen; Rodríguez, Pilar; Zúñiga, Paz; Reyes, Fernando

    2009-12-01

    Exploration of marine-derived actinomycetes as a source of antitumor compounds has led to the isolation of a new member of the tartrolon series, tartrolon D (4). This new compound was obtained from Streptomyces sp. MDG-04-17-069 fermentation broths and displayed strong cytotoxic activity against three human tumor cell lines. Additionally, the known compound ikarugamycin (5) was also found in the culture broths of the same microorganism. The structure of this new tartrolon was established by a combination of spectroscopic techniques (1D and 2D NMR, HRMS, and UV) as well as by comparison with published data for similar compounds.

  19. In Vitro and In Vivo Plant Growth Promoting Activities and DNA Fingerprinting of Antagonistic Endophytic Actinomycetes Associates with Medicinal Plants.

    Science.gov (United States)

    Passari, Ajit Kumar; Mishra, Vineet Kumar; Gupta, Vijai Kumar; Yadav, Mukesh Kumar; Saikia, Ratul; Singh, Bhim Pratap

    2015-01-01

    Endophytic actinomycetes have shown unique plant growth promoting as well as antagonistic activity against fungal phytopathogens. In the present study forty-two endophytic actinomycetes recovered from medicinal plants were evaluated for their antagonistic potential and plant growth-promoting abilities. Twenty-two isolates which showed the inhibitory activity against at least one pathogen were subsequently tested for their plant-growth promoting activities and were compared genotypically using DNA based fingerprinting, including enterobacterial repetitive intergenic consensus (ERIC) and BOX repetitive elements. Genetic relatedness based on both ERIC and BOX-PCR generates specific patterns corresponding to particular genotypes. Exponentially grown antagonistic isolates were used to evaluate phosphate solubilization, siderophores, HCN, ammonia, chitinase, indole-3-acetic acid production, as well as antifungal activities. Out of 22 isolates, the amount of indole-3-acetic acid (IAA) ranging between 10-32 μg/ml was produced by 20 isolates and all isolates were positive for ammonia production ranging between 5.2 to 54 mg/ml. Among 22 isolates tested, the amount of hydroxamate-type siderophores were produced by 16 isolates ranging between 5.2 to 36.4 μg/ml, while catechols-type siderophores produced by 5 isolates ranging from 3.2 to 5.4 μg/ml. Fourteen isolates showed the solubilisation of inorganic phosphorous ranging from 3.2 to 32.6 mg/100ml. Chitinase and HCN production was shown by 19 and 15 different isolates, respectively. In addition, genes of indole acetic acid (iaaM) and chitinase (chiC) were successively amplified from 20 and 19 isolates respectively. The two potential strains Streptomyces sp. (BPSAC34) and Leifsonia xyli (BPSAC24) were tested in vivo and improved a range of growth parameters in chilli (Capsicum annuum L.) under greenhouse conditions. This study is the first published report that actinomycetes can be isolated as endophytes from within these

  20. In Vitro and In Vivo Plant Growth Promoting Activities and DNA Fingerprinting of Antagonistic Endophytic Actinomycetes Associates with Medicinal Plants.

    Directory of Open Access Journals (Sweden)

    Ajit Kumar Passari

    Full Text Available Endophytic actinomycetes have shown unique plant growth promoting as well as antagonistic activity against fungal phytopathogens. In the present study forty-two endophytic actinomycetes recovered from medicinal plants were evaluated for their antagonistic potential and plant growth-promoting abilities. Twenty-two isolates which showed the inhibitory activity against at least one pathogen were subsequently tested for their plant-growth promoting activities and were compared genotypically using DNA based fingerprinting, including enterobacterial repetitive intergenic consensus (ERIC and BOX repetitive elements. Genetic relatedness based on both ERIC and BOX-PCR generates specific patterns corresponding to particular genotypes. Exponentially grown antagonistic isolates were used to evaluate phosphate solubilization, siderophores, HCN, ammonia, chitinase, indole-3-acetic acid production, as well as antifungal activities. Out of 22 isolates, the amount of indole-3-acetic acid (IAA ranging between 10-32 μg/ml was produced by 20 isolates and all isolates were positive for ammonia production ranging between 5.2 to 54 mg/ml. Among 22 isolates tested, the amount of hydroxamate-type siderophores were produced by 16 isolates ranging between 5.2 to 36.4 μg/ml, while catechols-type siderophores produced by 5 isolates ranging from 3.2 to 5.4 μg/ml. Fourteen isolates showed the solubilisation of inorganic phosphorous ranging from 3.2 to 32.6 mg/100ml. Chitinase and HCN production was shown by 19 and 15 different isolates, respectively. In addition, genes of indole acetic acid (iaaM and chitinase (chiC were successively amplified from 20 and 19 isolates respectively. The two potential strains Streptomyces sp. (BPSAC34 and Leifsonia xyli (BPSAC24 were tested in vivo and improved a range of growth parameters in chilli (Capsicum annuum L. under greenhouse conditions. This study is the first published report that actinomycetes can be isolated as endophytes from

  1. Cephamycins, a New Family of β-Lactam Antibiotics I. Production by Actinomycetes, Including Streptomyces lactamdurans sp. n1

    Science.gov (United States)

    Stapley, E. O.; Jackson, M.; Hernandez, S.; Zimmerman, S. B.; Currie, S. A.; Mochales, S.; Mata, J. M.; Woodruff, H. B.; Hendlin, D.

    1972-01-01

    A number of actinomycetes isolated from soil were found to produce one or more members of a new family of antibiotics, the cephamycins, which are structurally related to cephalosporin C. The cephamycins were produced in submerged fermentation in a wide variety of media by one or more of eight different species of Streptomyces, including a newly described species, S. lactamdurans. These antibiotics exhibit antibacterial activity against a broad spectrum of bacteria which includes many that are resistant to the cephalosporins and penicillins. PMID:4790552

  2. Research Progress of Active Metabolite of Marine Actinomycetes%海洋放线菌活性代谢产物的研究进展

    Institute of Scientific and Technical Information of China (English)

    宗志友; 魏刚; 斯聪聪; 王莹

    2011-01-01

    The research background of marine actinomycetes are briefly introduced in this paper. The research progresses of the active metabolite of marine actinomycetes in recent years are summarized and these metabolites are discussed individually according to their structures.%简要介绍了海洋放线菌的研究背景,重点对近年来发现的由海洋放线菌产生的活性代谢产物的研究进展作一概述,并对这些代谢产物依据其结构进行分类概述.

  3. Production and assay of cellulolytic enzyme activity of Enterobacter cloacae WPL 214 isolated from bovine rumen fluid waste of Surabaya abbatoir, Indonesia

    Directory of Open Access Journals (Sweden)

    W. P. Lokapirnasari

    2015-03-01

    Full Text Available Aim: This study aims to produce and assay cellulolytic enzyme activity (endo-(1,4-β-D-glucanase, exo-(1,4-β-Dglucanase, and β-glucosidase, at optimum temperature and optimum pH of Enterobacter cloacae WPL 214 isolated from bovine rumen fluid waste of Surabaya Abbatoir, Indonesia. Materials and Methods: To produce enzyme from a single colony of E. cloacae WPL 214, 98 × 1010 CFU/ml of isolates was put into 20 ml of liquid medium and incubated in a shaker incubator for 16 h at 35°C in accordance with growth time and optimum temperature of E. cloacae WPL 214. Further on, culture was centrifuged at 6000 rpm at 4°C for 15 min. Pellet was discarded while supernatant containing cellulose enzyme activity was withdrawn to assay endo-(1,4-β-D-glucanase, exo-(1,4-β-D-glucanase, and β-glucosidase. Results: Cellulase enzyme of E. cloacae WPL 214 isolates had endoglucanase activity of 0.09 U/ml, exoglucanase of 0.13 U/ml, and cellobiase of 0.10 U/ml at optimum temperature 35°C and optimum pH 5. Conclusion: E. cloacae WPL 214 isolated from bovine rumen fluid waste produced cellulose enzyme with activity as cellulolytic enzyme of endo-(1,4-β-D-glucanase, exo-(1,4-β-D-glucanase and β-glucosidase.

  4. Influence of an aerobic fungus grown on solid culture on ruminal degradability and on a mixture culture of anaerobic cellulolytic bacteria.

    Science.gov (United States)

    Hernández-Díaz, R; Pimentel-González, D J; Figueira, A C; Viniegra-González, G; Campos-Montiel, R G

    2010-06-01

    In this work, the effect of a solid fungal culture of Aspergillus niger (An) grown on coffee pulp on the in situ ruminal degradability (RD) of corn stover was evaluated. In addition, the effect of its extracts on the in vitro dry matter disappearance (IVDMD) and on a mixed culture of anaerobic cellulolytic bacteria (MCACB) was also investigated. The solid ferment was a crude culture of An, grown on coffee pulp. Regarding in situ RD, a significant difference (p < 0.05) was found between treatment with 200 g/day of the solid culture and control (no solid culture added) on dry matter, crude protein and neutral detergent fibre on RD. All the water extracts (pH 4, 7 and 10) enhanced IVDMD and stimulated the cellulolytic activity on a MCACB. Ultrafiltration results showed that active compounds with a molecular weight lower than 30 kDa were responsible for the effect on MCACB. Such results suggest that the effects of the solid An culture in RD are related to the presence of water soluble compounds having a molecular weight lower than 30 kDa.

  5. Effect of feeding palm oil by-products based diets on total bacteria, cellulolytic bacteria and methanogenic archaea in the rumen of goats.

    Directory of Open Access Journals (Sweden)

    Abdelrahim Abubakr

    Full Text Available Rumen microorganisms are responsible for digestion and utilization of dietary feeds by host ruminants. Unconventional feed resources could be used as alternatives in tropical areas where feed resources are insufficient in terms of quality and quantity. The objective of the present experiment was to evaluate the effect of diets based on palm oil (PO, decanter cake (DC or palm kernel cake (PKC on rumen total bacteria, selected cellulolytic bacteria, and methanogenic archaea. Four diets: control diet (CD, decanter cake diet (DCD, palm kernel cake diet (PKCD and CD plus 5% PO diet (CPOD were fed to rumen cannulated goats and rumen samples were collected at the start of the experimental diets (day 0 and on days 4, 6, 8, 12, 18, 24 and 30 post dietary treatments. Feeding DCD and PKCD resulted in significantly higher (P<0.05 DNA copy number of total bacteria, Fibrobacter succinogenes, Ruminococcus flavefeciens, and Ruminococcus albus. Rumen methanogenic archaea was significantly lower (P<0.05 in goats fed PKCD and CPOD and the trend showed a severe reduction on days 4 and 6 post experimental diets. In conclusion, results indicated that feeding DCD and PKC increased the populations of cellulolytic bacteria and decreased the density of methanogenic archaea in the rumen of goats.

  6. Effect of feeding palm oil by-products based diets on total bacteria, cellulolytic bacteria and methanogenic archaea in the rumen of goats.

    Science.gov (United States)

    Abubakr, Abdelrahim; Alimon, Abdul Razak; Yaakub, Halimatun; Abdullah, Norhani; Ivan, Michael

    2014-01-01

    Rumen microorganisms are responsible for digestion and utilization of dietary feeds by host ruminants. Unconventional feed resources could be used as alternatives in tropical areas where feed resources are insufficient in terms of quality and quantity. The objective of the present experiment was to evaluate the effect of diets based on palm oil (PO), decanter cake (DC) or palm kernel cake (PKC) on rumen total bacteria, selected cellulolytic bacteria, and methanogenic archaea. Four diets: control diet (CD), decanter cake diet (DCD), palm kernel cake diet (PKCD) and CD plus 5% PO diet (CPOD) were fed to rumen cannulated goats and rumen samples were collected at the start of the experimental diets (day 0) and on days 4, 6, 8, 12, 18, 24 and 30 post dietary treatments. Feeding DCD and PKCD resulted in significantly higher (P<0.05) DNA copy number of total bacteria, Fibrobacter succinogenes, Ruminococcus flavefeciens, and Ruminococcus albus. Rumen methanogenic archaea was significantly lower (P<0.05) in goats fed PKCD and CPOD and the trend showed a severe reduction on days 4 and 6 post experimental diets. In conclusion, results indicated that feeding DCD and PKC increased the populations of cellulolytic bacteria and decreased the density of methanogenic archaea in the rumen of goats.

  7. Screening of Actinomycetes from mangrove ecosystem for L-asparaginase activity and optimization by response surface methodology.

    Science.gov (United States)

    Usha, Rajamanickam; Mala, Krishnaswami Kanjana; Venil, Chidambaram Kulandaisamy; Palaniswamy, Muthusamy

    2011-01-01

    Marine actinomycetes were isolated from sediment samples collected from Pitchavaram mangrove ecosystem situated along the southeast coast of India. Maximum actinomycete population was noted in rhizosphere region. About 38% of the isolates produced L-asparaginase. One potential strain KUA106 produced higher level of enzyme using tryptone glucose yeast extract medium. Based on the studied phenotypic characteristics, strain KUA106 was identified as Streptomyces parvulus KUA106. The optimization method that combines the Plackett-Burman design, a factorial design and the response surface method, which were used to optimize the medium for the production of L-asparaginase by Streptomycetes parvulus. Four medium factors were screened from eleven medium factors by Plackett-Burman design experiments and subsequent optimization process to find out the optimum values of the selected parameters using central composite design was performed. Asparagine, tryptone, d) extrose and NaCl components were found to be the best medium for the L-asparaginase production. The combined optimization method described here is the effective method for screening medium factors as well as determining their optimum level for the production of L-asparaginase by Streptomycetes parvulus KUAP106.

  8. Diversity of culturable nocardioform actinomycetes from wastewater treatment plants in Spain and their role in the biodegradability of aromatic compounds.

    Science.gov (United States)

    Soler, Albert; García-Hernández, Jorge; Zornoza, Andrés; Alonso, José Luis

    2017-03-06

    Currently, municipal and industrial wastewater treatment plants (WWTPs) are mainly focusing on reduction of biological oxygen demand and on the removal of nutrients. However, there are microorganisms that interfere with the process. In this environment, there is a large diversity of microorganisms that have not been studied in detail and that could provide real and practical solutions to the foaming problems. Among such microorganisms, Gram-positive actinomycete bacteria are of special interest because they are known for producing secondary metabolites as well as chemically diverse compounds and for their capacity to degrade recalcitrant pollutants. Three different media were chosen to isolate actinomycetes from 28 WWTPs in Spain. A total of 189 activated sludge samples were collected; 126 strains were isolated and identified to belong to 1 suborder, i.e. Corynebacterineae, and 7 genera, i.e. Corynebacterium, Dietzia, Gordonia, Mycobacterium, Rhodococcus, Tsukamurella and Williamsia. Furthermore, 71 strains were capable of biodegrading at least 1 aromatic product, and that 27 of them amplified for catA gene. The results of this research help us understand the complexity of the foam-forming microbial populations in Spain and it shows that WWTPs can be a good source of microorganisms that can degrade phenol or naphthalene.

  9. Isolation and evaluation of proteolytic actinomycete isolates as novel inducers of pearl millet downy mildew disease protection

    Science.gov (United States)

    Jogaiah, Sudisha; Kurjogi, Mahantesh; Govind, Sharathchandra Ramasandra; Huntrike, Shekar Shetty; Basappa, Vedamurthy Ankala; Tran, Lam-Son Phan

    2016-01-01

    Native endophytic actinomycetes isolated from pearl millet roots were examined for their efficacy to protect pearl millet against downy mildew. Nineteen of 39 isolates were found to be proteolytic, of which 7 strains could directly suppress the sporangium formation of Sclerospora graminicola, the pearl millet downy mildew pathogen. Thus, mycelial suspensions containing either spores or cell-free extract of these 7 isolates were used for seed-coating and -soaking treatments to test for their induction of downy mildew resistance. Results indicated that seed-coating overall provided better protection to downy mildew than seed-soaking. In both treatments, the tested isolates demonstrated differential abilities in downy mildew disease protection, with Streptomyces griseus SJ_UOM-07-09 and Streptosporangium roseum SJ_UOM-18-09 showing the highest protection rates. Additionally, the levels of disease protection conferred by the actinomycetes were just slightly lower than that of the systemic fungicide Apron, suggesting their effectiveness. Further studies revealed that the more rapid root colonization by SJ_UOM-18-09 resulted in faster and higher induced resistance in comparison with SJ_UOM-07-09 under greenhouse conditions, indicating that SJ_UOM-18-09 was superior than SJ_UOM-07-09 in inducing resistance. Results from this study provide comprehensive information on biocontrol functions of SJ_UOM- 18-09 with great potential to control downy mildew disease in pearl millet. PMID:27499196

  10. Effect of crude extracts of selected actinomycetes on biofilm formation of A. schindleri, M. aci, and B. cereus.

    Science.gov (United States)

    Saleem, Hafiz Ghulam Murtaza; Aftab, Usman; Sajid, Imran; Abbas, Zaigham; Sabri, Anjum Nasim

    2015-05-01

    Actinomycetes are well known group of gram positive bacteria for their potential to produce antibiotics. This study sought to assess the ability of the selected actinomycetes to control biofilm forming bacteria isolated from different dental plaque samples. On the basis of morphological differences three out of ten different dental plaque bacterial isolates were selected for further study. These isolates were biochemically and genetically characterized and were identified as Acinetobacter schinndleri, Moraxella aci, and Bacillus cereus. Antibiotic resistant profile was measured through disc diffusion method and found that all three isolates were moderately sensitive to ofloxacin and erythromycin and resistant to trimethoprim. Antibacterial activity of ten different Streptomyces strains was assessed through an agar plug and well diffusion method against three dental biofilm forming bacteria. Two Streptomyces strains named as S. erythrogriseus and S. labedae showed good antibacterial activity against Moraxella and Acinetobacter strains. Ability of the four active antibiotic producing strains to inhibit biofilm formation was assessed using microtiter biofilm detection assay. It was found that biofilm forming ability of Acinetobacter and Moraxella was inhibited by S. labedae an antibiotic producing strain, while S. macrosporeus can only inhibit biofilm formation by B. cereus.

  11. Cytotoxicity of actinomycetes associated with the ascidian Eudistoma vannamei (Millar, 1977, endemic of northeastern coast of Brazil

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    Paula C Jimenez

    2013-04-01

    Full Text Available Previous studies demonstrated that the crude extract of the ascidian Eudistoma vannamei, endemic from northeasttern Brazil, strongly hinders growth of tumor cells in vitro by inducing apoptosis due to tryptophan derivatives, which are commonly found in bacteria. This study presents a bioactivity-guided screening among actinomycetes, associated with E. vannamei, aiming at recognizing active principles with biological relevance. Twenty strains of actinomycetes, designated as EVA 0101 through 0120, were isolated from colonies of E. vannamei among which 11 were selected for cytotoxicity evaluation. The extracts from EVA 0102, 0103, 0106, 0109 and 0113 were the most active, and were further studied for IC50 determination and chemical analysis by ¹H NMR. IC50 values obtained ranged from 3.62 µg mL-1 (for EVA 0109 in leukemia cells to 84.65 µg/mL (for EVA 0106 in melanoma cells. All active extracts exhibited the same TLC and spectroscopic profiles, suggesting the presence of quinones and other related secondary metabolites. Furthermore, these strains were identified and compared based on their respective 16S rRNA sequences. The results herein identified the five strains as Micromonospora spp. while phylogenetic analysis suggests that they are possibly two different Micromonospora species producing the cytotoxic compounds.

  12. Broad spectrum antimicrobial activity of forest-derived soil actinomycete, Nocardia sp. PB-52

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    Priyanka eSharma

    2016-03-01

    Full Text Available A mesophilic actinomycete strain designated as PB-52 was isolated from soil samples of Pobitora Wildlife Sanctuary of Assam, India. Based on phenotypic and molecular characteristics, the strain was identified as Nocardia sp. which shares 99.7% sequence similarity with Nocardia niigatensis IFM 0330 (NR_112195. The strain is a Gram-positive filamentous bacterium with rugose spore surface which exhibited a wide range of antimicrobial activity against Gram-positive bacteria including methicillin resistant Staphylococcus aureus (MRSA, Gram-negative bacteria and yeasts. Optimization for the growth and antimicrobial metabolite production of the strain PB-52 was carried out in batch culture under shaking condition. The optimum growth and the antimicrobial metabolite production by the strain PB-52 was recorded in GLM medium at 28ºC, initial pH 7.4 of the medium and incubation period of eight days. Based on polyketide synthases (PKS and nonribosomal peptide synthetases (NRPS gene-targeted PCR amplification, the occurrence of both of these biosynthetic pathways was detected which might be involved in the production of antimicrobial metabolite in PB-52. Extract of the fermented broth culture of PB-52 was prepared with organic solvent extraction method using ethyl acetate. The ethyl acetate extract of PB-52 (EA-PB-52 showed lowest minimum inhibitory concentration (MIC against Staphylococcus aureus MTCC 96 (0.975 μg/ml whereas highest was recorded against Klebsiella pneumoniae ATCC 13883 (62.5 μg/ml. Scanning electron microscopy (SEM revealed that treatment of the test microorganisms with EA-PB-52 destroyed the targeted cells with prominent loss of cell shape and integrity. In order to determine the constituents responsible for its antimicrobial activity, EA-PB-52 was subjected to chemical analysis using gas chromatography-mass spectrometry (GC-MS. GC-MS analysis showed the presence of twelve different chemical constituents in the extract, some of which

  13. Streptomyces luozhongensis sp. nov., a novel actinomycete with antifungal activity and antibacterial activity.

    Science.gov (United States)

    Zhang, Renwen; Han, Xiaoxue; Xia, Zhanfeng; Luo, Xiaoxia; Wan, Chuanxing; Zhang, Lili

    2017-02-01

    A novel actinomycete strain, designated TRM 49605(T), was isolated from a desert soil sample from Lop Nur, Xinjiang, north-west China, and characterised using a polyphasic taxonomic approach. The strain exhibited antifungal activity against the following strains: Saccharomyces cerevisiae, Curvularia lunata, Aspergillus flavus, Aspergillus niger, Fusarium oxysporum, Penicillium citrinum, Candida albicans and Candida tropicalis; Antibacterial activity against Bacillus subtilis, Staphylococcus epidermidis and Micrococcus luteus; and no antibacterial activity against Escherichia coli. Phylogenetic analysis based on 16S rRNA gene sequences affiliated strain TRM 49605(T) to the genus Streptomyces. Strain TRM 49605(T) shows high sequence similarities to Streptomyces roseolilacinus NBRC 12815(T) (98.62 %), Streptomyces flavovariabilis NRRL B-16367(T) (98.45 %) and Streptomyces variegatus NRRL B-16380(T) (98.45 %). Whole cell hydrolysates of strain TRM 49605(T) were found to contain LL-diaminopimelic acid as the diagnostic diamino acid and galactose, glucose, xylose and mannose as the major whole cell sugars. The major fatty acids in strain TRM 49605(T) were identified as iso C16:0, anteiso C15:0, C16:0 and Summed Feature 5 as defined by MIDI. The main menaquinones were identified as MK-9(H4), MK-9(H6), MK-9(H8) and MK-10(H6). The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol and phosphatidylinositol mannoside. The G+C content of the genomic DNA was determined to be 71.2 %. The DNA-DNA relatedness between strain TRM 49605(T) and the phylogenetically related strain S. roseolilacinus NBRC 12815(T) was 60.12 ± 0.06 %, which is lower than the 70 % threshold value for delineation of genomic prokaryotic species. Based on the phenotypic, chemotaxonomic and phylogenetic data, strain TRM 49605(T) (=CCTCC AA2015026(T) = KCTC 39666(T)) should be designated as the type strain of a novel species of the genus

  14. Pharmaceutical composition to protect an animal against a disorder arising from an infection with a bacterium that belongs to the group of nocardioform actinomycetes

    NARCIS (Netherlands)

    Arnoldus, Christiaan Jacobs; van der Geize, Robert; Dijkhuizen, Lubbert

    2011-01-01

    The invention pertains to a pharmaceutical composition to protect an animal against a disorder arising from an infection with a bacterium that belongs to the group of nocardioform actinomycetes having the ability to survive within macrophages of the animal, comprising live bacteria of a nocardioform

  15. XML In Vitro Comparison of MIC Crude Extracts of Active Actinomycetes Isolated with Terbinafine, Griseofulvin Ketoconazole and Fluconazole against Microsporum Canis, Microsporum Gypseum and Trichophyton Mentagrophytes

    Directory of Open Access Journals (Sweden)

    Keikha, N. (MSc

    2015-05-01

    Full Text Available Background and Objective: Dermatophytes are the fungi that have the ability to attack the keratinized tissues such as the skin, hair and nails. Infections caused by these organisms are named dermatophytosis. We aimed to compare Minimum inhibitory concentration (MIC of Crude extracts of Active Actinomycete Isolates with Terbinafine, Griseofulvin, Ketoconazole and Fluconazole Drugs against Microsporum Canis, Microsporum gypseum and Trichophyton mentagrophytes. Material and Methods: In this experimental study, in order to find MIC by actionmycete, 100 isolates were studied and then crude extracts of the active actinomycete isolates were prepared in sterile conditions. Finally, the crude extracts obtained at different concentrations were used to obtain the MIC of Microsporum Canis, Microsporum gypseum and Trichophyton mentagrophytes. Moreover, various concentrations of the drugs such as terbinafine, griseofulvin, ketoconazole and fluconazole in solvent Dimethyl sulfoxide (DMSO were prepared and their growth inhibitory effect was evaluated and then compared with the results obtained from the crude extract of active actinomycete isolates. Results: the crude extracts obtained from active Actioiomycetes isolates and the drugs such as terbinafine, griseofulvin, ketoconazole and fluconazole, in a dose-dependent manner, could inhibit the growth of Microsporum Canis, Microsporum gypseum and Trichophyton Mentagrophytes. Conclusion: compared to MIC of Crude extract of active actinomycete isolates, Terbinafine has a significant effect on the growth inhibition in all of the fungal Dermatophytes and then griseofulvin, ketoconazole and fluconazole are in the next rank, respectively.

  16. Pharmaceutical composition to protect an animal against a disorder arising from an infection with a bacterium that belongs to the group of nocardioform actinomycetes

    NARCIS (Netherlands)

    Arnoldus, Christiaan Jacobs; van der Geize, Robert; Dijkhuizen, Lubbert

    2011-01-01

    The invention pertains to a pharmaceutical composition to protect an animal against a disorder arising from an infection with a bacterium that belongs to the group of nocardioform actinomycetes having the ability to survive within macrophages of the animal, comprising live bacteria of a nocardioform

  17. Complete Genome Sequence of Micromonospora Strain L5, a Potential Plant-Growth-Regulating Actinomycete, Originally Isolated from Casuarina equisetifolia Root Nodules

    Energy Technology Data Exchange (ETDEWEB)

    Hirsch, A. M.; Alvarado, J.; Bruce, D.; Chertkov, O.; De Hoff, P. L.; Detter, J. C.; Fujishige, N. A.; Goodwin, L. A.; Han, J.; Han, S.; Ivanova, N.; Land, M. L.; Lum, M. R.; Milani-Nejad, N.; Nolan, M.; Pati, A.; Pitluck, S.; Tran, S. S.; Woyke, T.; Valdes, M.

    2013-08-29

    Micromonospora species live in diverse environments and exhibit a broad range of functions including antibiotic production, biocontrol, and ability to degrade complex polysaccharides. To learn more about these versatile actinomycetes, we sequenced the genome of strain L5, originally isolated from root nodules of an actinorhizal plant growing in Mexico.

  18. The genome sequences of Cellulomonas fimi and "Cellvibrio gilvus" reveal the cellulolytic strategies of two facultative anaerobes, transfer of "Cellvibrio gilvus" to the genus Cellulomonas, and proposal of Cellulomonas gilvus sp. nov.

    Directory of Open Access Journals (Sweden)

    Melissa R Christopherson

    Full Text Available Actinobacteria in the genus Cellulomonas are the only known and reported cellulolytic facultative anaerobes. To better understand the cellulolytic strategy employed by these bacteria, we sequenced the genome of the Cellulomonas fimi ATCC 484(T. For comparative purposes, we also sequenced the genome of the aerobic cellulolytic "Cellvibrio gilvus" ATCC 13127(T. An initial analysis of these genomes using phylogenetic and whole-genome comparison revealed that "Cellvibrio gilvus" belongs to the genus Cellulomonas. We thus propose to assign "Cellvibrio gilvus" to the genus Cellulomonas. A comparative genomics analysis between these two Cellulomonas genome sequences and the recently completed genome for Cellulomonas flavigena ATCC 482(T showed that these cellulomonads do not encode cellulosomes but appear to degrade cellulose by secreting multi-domain glycoside hydrolases. Despite the minimal number of carbohydrate-active enzymes encoded by these genomes, as compared to other known cellulolytic organisms, these bacteria were found to be proficient at degrading and utilizing a diverse set of carbohydrates, including crystalline cellulose. Moreover, they also encode for proteins required for the fermentation of hexose and xylose sugars into products such as ethanol. Finally, we found relatively few significant differences between the predicted carbohydrate-active enzymes encoded by these Cellulomonas genomes, in contrast to previous studies reporting differences in physiological approaches for carbohydrate degradation. Our sequencing and analysis of these genomes sheds light onto the mechanism through which these facultative anaerobes degrade cellulose, suggesting that the sequenced cellulomonads use secreted, multidomain enzymes to degrade cellulose in a way that is distinct from known anaerobic cellulolytic strategies.

  19. Isolation, abundance and phylogenetic affiliation of endophytic actinomycetes associated with medicinal plants and screening for their in vitro antimicrobial biosynthetic potential

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    Ajit Kumar Passari

    2015-04-01

    Full Text Available Microorganisms associated with medicinal plants are of interest as the producers of important bioactive compounds. To date, the diversity of culturable endophytic actinomycetes associated with medicinal plants is in its initial phase of exploration. In this study, 42 endophytic actinomycetes were isolated from different organs of seven selected medicinal plants. The highest number of isolates (n=22, 52.3% of actinomycetes was isolated from roots, followed by stems (n=9, 21.4%, leaves (n=6, 14.2%, flowers (n=3, 7.1% and petioles (n=2, 4.7%. The genus Streptomyces was the most dominant among the isolates (66.6% in both the locations (Dampa TRF and Phawngpuii NP, Mizoram, India. From a total of 42 isolates, 22 isolates were selected for further studies based on their ability to inhibit one of the tested human bacterial or fungal pathogen. Selected isolates were identified based on 16S rRNA gene analysis and subsequently the isolates were grouped to four different genera; Streptomyces, Brevibacterium, Microbacterium and Leifsonia. Antibiotic sensitivity assay was performed to understand the responsible antimicrobials present in the isolates showing the antimicrobial activities and revealed that the isolates were mostly resistant to penicillin G and ampicillin. Further, antimicrobial properties and antibiotic sensitivity assay in combination with the results of amplification of biosynthetic genes polyketide synthase (PKS-I and nonribosomal peptide synthetase (NRPS showed that the endophytic actinomycetes associated with the selected medicinal plants have broad-spectrum antimicrobial activity. This is the first report of the isolation of Brevibacterium sp., Microbacterium sp. and Leifsonia xyli from endophytic environments of medicinal plants, Mirabilis jalapa and Clerodendrum colebrookianum. Our results emphasize that endophytic actinomycetes associated with medicinal plants are an unexplored resource for the discovery of biologically active

  20. Antibiotics Resistance Profiling and In-Vitro Inhibition of Clinical Klebsiella Strains by Actinomycetes Isolated From Different Ecological Niches in Pakistan

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    Noureen

    2016-02-01

    Full Text Available Background Multidrug resistance among different pathogens is increasing immensely day by day. To control these problems, we need new potent antimicrobial agents in repository of antibiotics. Objectives This study aimed at investigation of antibiotics resistance pattern of pathogenic Klebsiella strains isolated from clinical samples in Lahore region Pakistan and study inhibition of resistant strains by natural extracts obtained from actinomycetes isolated from different ecological niches in Pakistan. Materials and Methods The isolated Klebsiella strains were identified by morphological, biochemical and physiological characterization along with 16S rRNA gene sequencing. Antibiotics susceptibility was determined by standard Kirby Bauer disc diffusion assay. The biological and chemical screening was performed for detection of active secondary metabolites produced by actinomycetes against resistant Klebsiella strains. Biological screenings include antimicrobial activity by agar diffusion assay and brine shrimp microwell cytotoxicity assay. In chemical screening, the crude extracts of actinomycetes strains were analysed by TLC and HPLC-UV techniques. Results The isolated Klebsiella strains showed resistance against most of the antibiotics as follows; ceftriaxone > cephalexin > cefpirome > cefoxitin = cefepime > levofloxacin > ciprofloxacin = ceftrazidime = fusidum > cefoperazone > ampicillin sulbactam. The actinomycetes strain A19, A20, A2, A10, A6 and A8 exhibited remarkable activity against resistant Klebsiella strains. The strains A19 and 20 showed excellent inhibitory effects on all isolated multidrug resistant Klebsiella strains. Conclusions The clinical Klebsiella strains isolated from Lahore region, Pakistan exhibited resistance to most commonly used antibiotics, which can be a serious threat to public health. The study reported some potential actinomycetes strains, which exhibit promising activity against multidrug resistant Klebsiella strains.

  1. Metagenomic of Actinomycetes Based on 16S rRNA and nifH Genes in Soil and Roots of Four Indonesian Rice Cultivars Using PCR-DGGE

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    Mahyarudin

    2015-07-01

    Full Text Available The research was conducted to study the metagenomic of actinomycetes based on 16S ribosomal RNA (rRNA and bacterial nifH genes in soil and roots of four rice cultivars. The denaturing gradient gel electrophoresis profile based on 16S rRNA gene showed that the diversity of actinomycetes in roots was higher than soil samples. The profile also showed that the diversity of actinomycetes was similar in four varieties of rice plant and three types of agroecosystem. The profile was partially sequenced and compared to GenBank database indicating their identity with closely related microbes. The blast results showed that 17 bands were closely related ranging from 93% to 100% of maximum identity with five genera of actinomycetes, which is Geodermatophilus, Actinokineospora, Actinoplanes, Streptomyces and Kocuria. Our study found that Streptomyces species in soil and roots of rice plants were more varied than other genera, with a dominance of Streptomyces alboniger and Streptomyces acidiscabies in almost all the samples. Bacterial community analyses based on nifH gene denaturing gradient gel electrophoresis showed that diversity of bacteria in soils which have nifH gene was higher than that in rice plant roots. The profile also showed that the diversity of those bacteria was similar in four varieties of rice plant and three types of agroecosystem. Five bands were closely related with nifH gene from uncultured bacterium clone J50, uncultured bacterium clone clod-38, and uncultured bacterium clone BG2.37 with maximum identity 99%, 98%, and 92%, respectively. The diversity analysis based on 16S rRNA gene differed from nifH gene and may not correlate with each other. The findings indicated the diversity of actinomycetes and several bacterial genomes analyzed here have an ability to fix nitrogen in soil and roots of rice plant.

  2. An approach to mitigating soil CO2 emission by biochemically inhibiting cellulolytic microbial populations through mediation via the medicinal herb Isatis indigotica

    Science.gov (United States)

    Wu, Hong-Sheng; Chen, Su-Yun; Li, Ji; Liu, Dong-Yang; Zhou, Ji; Xu, Ya; Shang, Xiao-Xia; Wei, Dong-yang; Yu, Lu-ji; Fang, Xiao-hang; Li, Shun-yi; Wang, Ke-ke

    2017-06-01

    Greenhouse gases (GHGs, particularly carbon dioxide (CO2)) emissions from soil under wheat production are a significant source of agricultural carbon emissions that have not been mitigated effectively. A field experiment and a static incubation study in a lab were conducted to stimulate wheat growth and investigate its potential to reduce CO2 emissions from soil through intercropping with a traditional Chinese medicinal herb called Isatis indigotica. This work was conducted by adding I. indigotica root exudates based on the quantitative real-time PCR (qPCR) analysis of the DNA copy number of the rhizosphere or bulk soil microbial populations. This addition was performed in relation to the CO2 formation by cellulolytic microorganisms (Penicillium oxalicum, fungi and Ruminococcus albus) to elucidate the microbial ecological basis for the molecular mechanism that decreases CO2 emissions from wheat fields using I. indigotica. The results showed that the panicle weight and full grains per panicle measured through intercropping with I. indigotica (NPKWR) increased by 39% and 28.6%, respectively, compared to that of the CK (NPKW). Intercropping with I. indigotica significantly decreased the CO2 emissions from soil under wheat cultivation. Compared with CK, the total CO2 emission flux during the wheat growth period in the I. indigotica (NPKWR) intercropping treatment decreased by 29.26%. The intensity of CO2 emissions per kg of harvested wheat grain declined from 7.53 kg CO2/kg grain in the NPKW (CK) treatment to 5.55 kg CO2/kg grain in the NPKWR treatment. The qPCR analysis showed that the DNA copy number of the microbial populations of cellulolytic microorganisms (P. oxalicum, fungi and R. albus) in the field rhizosphere around I. indigotica or in the bulk soil under laboratory incubation was significantly lower than that of CK. This finding indicated that root exudates from I. indigotica inhibited the activity and number of cellulolytic microbial populations, which led

  3. Investigations on potato pulp as a dietary fiber source. The influence of pectolytic and cellulolytic enzymes. Untersuchungen an Kartoffelpuelpe als Ballaststoffquelle. Zum Einfluss von pektolytischen und cellulolytischen Enzymen

    Energy Technology Data Exchange (ETDEWEB)

    Dongowski, G. (Deutsches Inst. fuer Ernaehrungsforschung Potsdam-Rehbruecke, Bergholz-Rehbruecke (Germany))

    1993-05-01

    The influence of treatment with pectolytic and cellulolytic enzyme preparations was investigated with reference to the separation of water and the composition of potato pulp. In contrast to pectinesterase, pectate lyase or cellulase it was found an intensive action on the pulp after incubation with Pectinex Ultra SP-L or pectinase/cellulase combinations. The content of pectin, starch and protein as well as the water binding capacity are varied in dependence of the used enzyme preparations. The occurring changes in the supermolecular structure of the potato pulp tissue are investigated by scanning electron microscopy. The grown biological structure is partly or extensive destroyed especially after action of pectinases and cellulases. The content of starch in the potato pulp preparations remains relatively high even after intensive treatment with cell wall degrading enzymes. (orig.)

  4. Anaerobic cellulolytic rumen fungal populations in goats fed with and without Leucaena leucocephala hybrid, as determined by real-time PCR.

    Science.gov (United States)

    Kok, Ching Mun; Sieo, Chin Chin; Tan, Hui Yin; Saad, Wan Zuhainis; Liang, Juan Boo; Ho, Yin Wan

    2013-10-01

    The effect of Leucaena leucocephala hybrid-Bahru (LLB), which contains a high concentration of condensed tannins, on cellulolytic rumen fungal population in goats was investigated using real-time PCR. The fungal population in goats fed LLB was inhibited during the first 10 days of feeding, but after 15 days of feeding, there was a tremendous increase of fungal population (157.0 μg/ml), which was about fourfold more than that in control goats (39.7 μg/ml). However, after this period, the fungal population decreased continuously, and at 30 days of feeding, the fungal population (50.6 μg/ml) was not significantly different from that in control goats (55.4 μg/ml).

  5. Activity-based protein profiling of secreted cellulolytic enzyme activity dynamics in Trichoderma reesei QM6a, NG14, and RUT-C30

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, Lindsey N.; Culley, David E.; Hofstad, Beth A.; Chauvigne-Hines, Lacie M.; Zink, Erika M.; Purvine, Samuel O.; Smith, Richard D.; Callister, Stephen J.; Magnuson, Jon M.; Wright, Aaron T.

    2013-12-01

    Development of alternative, non-petroleum based sources of bioenergy that can be applied in the short-term find great promise in the use of highly abundant and renewable lignocellulosic plant biomass.1 This material obtained from different feedstocks, such as forest litter or agricultural residues, can yield liquid fuels and other chemical products through biorefinery processes.2 Biofuels are obtained from lignocellulosic materials by chemical pretreatment of the biomass, followed by enzymatic decomposition of cellulosic and hemicellulosic compounds into soluble sugars that are converted to desired chemical products via microbial metabolism and fermentation.3, 4 To release soluble sugars from polymeric cellulose multiple enzymes are required, including endoglucanase, exoglucanase, and β-glucosidase.5, 6 However, the enzymatic hydrolysis of cellulose into soluble sugars remains a significant limiting factor to the efficient and economically viable utilization of lignocellulosic biomass for transport fuels.7, 8 The primary industrial source of cellulose and hemicellulases is the mesophilic soft-rot fungus Trichoderma reesei,9 having widespread applications in food, feed, textile, pulp, and paper industries.10 The genome encodes 200 glycoside hydrolases, including 10 cellulolytic and 16 hemicellulolytic enzymes.11 The hypercellulolytic catabolite derepressed strain RUT-C30 was obtained through a three-step UV and chemical mutagenesis of the original T. reesei strain QM6a,12, 13 in which strains M7 and NG14 were intermediate, having higher cellulolytic activity than the parent strain but less activity and higher catabolite repression than RUT-C30.14 Numerous methods have been employed to optimize the secreted enzyme cocktail of T. reesei including cultivation conditions, operational parameters, and mutagenesis.3 However, creating an optimal and economical enzyme mixture for production-scale biofuels synthesis may take thousands of experiments to identify.

  6. Kallotenue papyrolyticum gen. nov., sp. nov., a cellulolytic and filamentous thermophile that represents a novel lineage (Kallotenuales ord. nov., Kallotenuaceae fam. nov.) within the class Chloroflexia

    Energy Technology Data Exchange (ETDEWEB)

    Cole, Jesse; Gieler, Brandon; Heisler, Devon; Palisoc, Maryknoll; Williams, Amanda; Dohnalkova, Alice; Ming, Hong; Yu, Tian T.; Dodsworth, Jeremy A.; Li, Wen J.; Hedlund, Brian P.

    2013-08-15

    Several closely-related, thermophilic, and cellulolytic bacterial strains, designated JKG1T, JKG2, JKG3, JKG4, and JKG5, were isolated from a cellulolytic enrichment (corn stover) incubated in the water column of Great Boiling Spring, NV. Strain JKG1T had cells of a diameter of 0.7 - 0.9 μm and length of ~2.0 μm that formed non-branched multicellular filaments reaching >300 μm. Spores were not formed and dense liquid cultures were red. The temperature range for growth was 45-65 °C, with an optimum of 55 °C. The pH range for growth was 5.6-9.0, with an optimum of 7.5. JKG1T grew as an aerobic heterotroph, utilizing glucose, sucrose, xylose, arabinose, cellobiose, carboxymethylcellulose, filter paper, microcrystalline cellulose, xylan, starch, casamino acids, tryptone, peptone, yeast extract, acetate, citrate, lactate, pyruvate, and glycerol as sole carbon sources, and was not observed to photosynthesize. The cells stained Gram-negative. Phylogenetic analysis using 16S rRNA gene sequences placed the new isolates in the class Chloroflexia, but distant from other cultivated members, with the highest sequence identity of 82.5% to Roseiflexus castenholzii. The major quinone was menaquinone-9; no ubiquinones were detected. The major cellular fatty acids (>5%) were C18:0, anteiso-C17:0, iso-C18:0, and iso-C17:0. C16:0, iso-C16:0, and C17:0. The peptidoglycan amino acids were alanine, ornithine, glutamic acid, serine, and asparagine. Whole-cell sugars included mannose, rhamnose, glucose, galactose, ribose, arabinose, and xylose. Morphological, phylogenetic, and chemotaxonomic results suggest that JKG1T is representative of a new lineage within the class Chloroflexia, which we propose to designate Kallotenue papyrolyticum gen. nov., sp. nov., Kallotenuaceae fam. nov., Kallotenuales ord. nov.

  7. Effects of Neutral Detergent Soluble Fiber and Sucrose Supplementation on Ruminal Fermentation, Microbial Synthesis, and Populations of Ruminal Cellulolytic Bacteria Using the Rumen Simulation Technique (RUSITEC)

    Institute of Scientific and Technical Information of China (English)

    ZHAO Xiang-hui; LIU Chan-juan; LI Chao-yun; YAO Jun-hu

    2013-01-01

    We evaluated the effects of neutral detergent soluble fiber (NDSF) and sucrose supplementation on ruminal fermentation, microbial synthesis, and populations of ruminal cellulolytic bacteria using the rumen simulation technique (RUSITEC). The experiment had a 2×2 factorial design with two dosages of sucrose, low (ca. 0.26 g d-1, low-sucrose) and high (ca. 1.01 g d-1, high-sucrose), and two dosages of supplied NDSF, low (1.95 g d-1, low-NDSF) and high (2.70 g d-1, high-NDSF). Interactions between NDSF and sucrose were detected for xylanase activity from solid fraction and apparent disappearance of neutral detergent fiber (NDF) and hemicellulose, with the lowest values observed for high-NDSF and high-sucrose treatment. Supplemental NDSF appeared to increase the molar proportion of acetate and reduce that of butyrate;however, the effects of supplemental sucrose on VFA profiles depended upon NDSF amount. There was a NDSF×sucrose interaction for the production of methane. High-NDSF fermenters had lower ammonia-N production, greater daily N flow of solid-associated microbial pellets and total microorganisms, and greater microbial synthesis efficiency compared with low-NDSF fermenters. Supplementation with NDSF resulted in an increase in 16S rDNA copies of Ruminococcus flavefaciens and a reduction in copies of Ruminococcus albus. Supplementation with sucrose tended to increase the 16S rDNA copies of R. albus from liquid fraction, but did not affect daily total microbial N flow and cellulolytic bacterium populations from solid fraction. These data indicate that the effects of the interaction between NDSF and sugars on ruminal fermentation and fiber digestion should be taken into account in diet formulation. Ruminal fermentation and metabolism of sugars warrant further investigation.

  8. Enhanced biomethane production rate and yield from lignocellulosic ensiled forage ley by in situ anaerobic digestion treatment with endogenous cellulolytic enzymes.

    Science.gov (United States)

    Speda, Jutta; Johansson, Mikaela A; Odnell, Anna; Karlsson, Martin

    2017-01-01

    Enzymatic treatment of lignocellulosic material for increased biogas production has so far focused on pretreatment methods. However, often combinations of enzymes and different physicochemical treatments are necessary to achieve a desired effect. This need for additional energy and chemicals compromises the rationale of using enzymes for low energy treatment to promote biogas production. Therefore, simpler and less energy intensive in situ anaerobic digester treatment with enzymes is desirable. However, investigations in which exogenous enzymes are added to treat the material in situ have shown mixed success, possibly because the enzymes used originated from organisms not evolutionarily adapted to the environment of anaerobic digesters. In this study, to examine the effect of enzymes endogenous to methanogenic microbial communities, cellulolytic enzymes were instead overproduced and collected from a dedicated methanogenic microbial community. By this approach, a solution with very high endogenous microbial cellulolytic activity was produced and tested for the effect on biogas production from lignocellulose by in situ anaerobic digester treatment. Addition of enzymes, endogenous to the environment of a mixed methanogenic microbial community, to the anaerobic digestion of ensiled forage ley resulted in significantly increased rate and yield of biomethane production. The enzyme solution had an instant effect on more readily available cellulosic material. More importantly, the induced enzyme solution also affected the biogas production rate from less accessible cellulosic material in a second slower phase of lignocellulose digestion. Notably, this effect was maintained throughout the experiment to completely digested lignocellulosic substrate. The induced enzyme solution collected from a microbial methanogenic community contained enzymes that were apparently active and stable in the environment of anaerobic digestion. The enzymatic activity had a profound effect on the

  9. Structures and comparative characterization of biosynthetic gene clusters for cyanosporasides, enediyne-derived natural products from marine actinomycetes.

    Science.gov (United States)

    Lane, Amy L; Nam, Sang-Jip; Fukuda, Takashi; Yamanaka, Kazuya; Kauffman, Christopher A; Jensen, Paul R; Fenical, William; Moore, Bradley S

    2013-03-20

    Cyanosporasides are marine bacterial natural products containing a chlorinated cyclopenta[a]indene core of suspected enediyne polyketide biosynthetic origin. Herein, we report the isolation and characterization of novel cyanosporasides C-F (3-6) from the marine actinomycetes Salinispora pacifica CNS-143 and Streptomyces sp. CNT-179, highlighted by the unprecedented C-2' N-acetylcysteamine functionalized hexose group of 6. Cloning, sequencing, and mutagenesis of homologous ~50 kb cyanosporaside biosynthetic gene clusters from both bacteria afforded the first genetic evidence supporting cyanosporaside's enediyne, and thereby p-benzyne biradical, biosynthetic origin and revealed the molecular basis for nitrile and glycosyl functionalization. This study provides new opportunities for bioengineering of enediyne derivatives and expands the structural diversity afforded by enediyne gene clusters.

  10. Creation of an HDAC-based yeast screening method for evaluation of marine-derived actinomycetes: discovery of streptosetin A.

    Science.gov (United States)

    Amagata, Taro; Xiao, Jing; Chen, Yi-Pei; Holsopple, Nicholas; Oliver, Allen G; Gokey, Trevor; Guliaev, Anton B; Minoura, Katsuhiko

    2012-12-28

    A histone deacetylase (HDAC)-based yeast assay employing a URA3 reporter gene was applied as a primary screen to evaluate a marine-derived actinomycete extract library and identify human class III HDAC (SIRT) inhibitors. On the basis of the bioassay-guided purification, a new compound designated as streptosetin A (1) was obtained from one of the active strains identified through the yeast assay. The gross structure of the new compound was elucidated from the 1D and 2D NMR data. The absolute stereostructure of 1 was determined based on X-ray crystal structure analysis and simulation of ECD spectra using time-dependent density functional theory calculations. This compound showed weak inhibitory activity against yeast Sir2p and human SIRT1 and SIRT2.

  11. Cultivable actinomycetes from rhizosphere of birch (Betula pendula) growing on a coal mine dump in Silets, Ukraine.

    Science.gov (United States)

    Ostash, Bohdan; Gren, Tetiana; Hrubskyy, Yaroslav; Tistechok, Stepan; Beshley, Stepan; Baranov, Volodymyr; Fedorenko, Victor

    2014-08-01

    Five actinomycete strains were isolated from the rhizosphere of birch, one of a few native tree forms capable of thriving on the upper level of a coal mine dump near the village of Silets (Lvivska region, Ukraine). No such strains were isolated from surrounding gangue, or from nearby grass Calamagrostis epigeios. Using 16S rDNA sequencing and analysis of cell wall aminoacids, four of these strains were shown to belong to genus Streptomyces and one to be Amycolatopsis. The isolates were able to produce siderophores and antibacterial compounds. In comparison to the reference strain Streptomyces coelicolor M145, certain rhizospheric isolates displayed somewhat increased survival in the presence of copper, iron(III), or chromium(VI) salts. The Amycolatopsis isolate was also shown to accumulate significant quantities of heavy metals from waste extracts. Possible roles of the described strains in coal mine dump ecology are discussed.

  12. Production and characterization of lipopeptide biosurfactant by a sponge-associated marine actinomycetes Nocardiopsis alba MSA10.

    Science.gov (United States)

    Gandhimathi, R; Seghal Kiran, G; Hema, T A; Selvin, Joseph; Rajeetha Raviji, T; Shanmughapriya, S

    2009-10-01

    A sponge-associated marine actinomycetes Nocardiopsis alba MSA10 was screened and evaluated for the production of biosurfactant. Biosurfactant production was confirmed by conventional screening methods including hemolytic activity, drop collapsing test, oil displacement method, lipase production and emulsification index. The active compound was extracted with three solvents including ethyl acetate, diethyl ether and dichloromethane. The diethyl ether extract was fractionated by TLC and semi-preparative HPLC to isolate the pure compound. In TLC, a single discrete spot was obtained with the R (f) 0.60 and it was extrapolated as valine. Based on the chemical characterization, the active compound was partially confirmed as lipopeptide. The optimum production was attained at pH 7, temperature 30 degrees C, and 1% salinity with glucose and peptone supplementation as carbon and nitrogen sources, respectively. Considering the biosurfactant production potential of N. alba, the strain could be developed for large-scale production of lipopeptide biosurfactant.

  13. Characterization and phylogenetic analysis of novel polyene type antimicrobial metabolite producing actinomycetes from marine sediments:Bay of Bengal India

    Institute of Scientific and Technical Information of China (English)

    Valan Arasu M; Asha KRT; Duraipandiyan V; Ignacimuthu S; Agastian P

    2012-01-01

    To isolate and indentify the promising antimicrobial metabolite producingStreptomyces strains from marine sediment samples from Andrapradesh coast of India. Methods:Antagonistic actinomycetes were isolated by starch casein agar medium and modified nutrient agar medium with 1% glucose used as a base for primary screening. Significant antimicrobial metabolite producing strains were selected and identified by using biochemical and 16S rDNA level. Minimum inhibitory concentrations of the organic extracts were done by using broth micro dilution method. Results: Among the 210 actinomycetes, 64.3% exhibited activity against Gram positive bacteria, 48.5 % showed activity towards Gram negative bacteria, 38.8% exhibited both Gram positive and negative bacteria and 80.85 % isolates revealed significant antifungal activity. However, five isolates AP-5, AP-18, AP-41 and AP-70 showed significant antimicrobial activity. The analysis of cell wall hydrolysates showed the presence of LL-diaminopimelic acid and glycine in all the isolates. Sequencing analysis indicated that the isolates shared 98.5%-99.8%sequence identity to the 16S rDNA gene sequences of the Streptomyces taxons. The antimicrobial substances were extracted using hexane and ethyl acetate from spent medium in which strains were cultivated at 30℃for five days. The antimicrobial activity was assessed using broth micro dilution technique. Each of the culture extracts from these five strains showed a typical polyene-like property. The lowest minimum inhibitory concentrations of ethyl acetate extracts against Escherichia coli and Curvularia lunata were 67.5 and 125.0 μg/mL, respectively. Conclusions: It can be concluded that hexane and ethyl acetate soluble extracellular products of novel isolates are effective against pathogenic bacteria and fungi.

  14. Characterization and phylogenetic analysis of novel polyene type antimicrobial metabolite producing actinomycetes from marine sediments: Bay of Bengal, India.

    Science.gov (United States)

    Valan, Arasu M; Asha, K R T; Duraipandiyan, V; Ignacimuthu, S; Agastian, P

    2012-10-01

    To isolate and indentify the promising antimicrobial metabolite producing Streptomyces strains from marine sediment samples from Andrapradesh coast of India. Antagonistic actinomycetes were isolated by starch casein agar medium and modified nutrient agar medium with 1% glucose used as a base for primary screening. Significant antimicrobial metabolite producing strains were selected and identified by using biochemical and 16S rDNA level. Minimum inhibitory concentrations of the organic extracts were done by using broth micro dilution method. Among the 210 actinomycetes, 64.3% exhibited activity against Gram positive bacteria, 48.5 % showed activity towards Gram negative bacteria, 38.8% exhibited both Gram positive and negative bacteria and 80.85 % isolates revealed significant antifungal activity. However, five isolates AP-5, AP-18, AP-41 and AP-70 showed significant antimicrobial activity. The analysis of cell wall hydrolysates showed the presence of LL-diaminopimelic acid and glycine in all the isolates. Sequencing analysis indicated that the isolates shared 98.5%-99.8% sequence identity to the 16S rDNA gene sequences of the Streptomyces taxons. The antimicrobial substances were extracted using hexane and ethyl acetate from spent medium in which strains were cultivated at 30°Cfor five days. The antimicrobial activity was assessed using broth micro dilution technique. Each of the culture extracts from these five strains showed a typical polyene-like property. The lowest minimum inhibitory concentrations of ethyl acetate extracts against Escherichia coli and Curvularia lunata were 67.5 and 125.0 µg/mL, respectively. It can be concluded that hexane and ethyl acetate soluble extracellular products of novel isolates are effective against pathogenic bacteria and fungi.

  15. Screening and Identifying of Antagonistic Actinomycetes against Ralstonia Solancearum%烟草青枯菌拮抗放线菌的筛选及鉴定

    Institute of Scientific and Technical Information of China (English)

    陆铮铮; 彭丽娟; 丁海霞; 左希; 彭杰; 蒋选利

    2013-01-01

    In order to select effective antagonistic actinomycetes against Ralstonia solanacearum, we isolated 56 strains of actinomycetes from healthy tobacco rhizosphere soils. By pairing culture on plate method to screen antagonistic actinomycetes, we attained 3 strains of antagonistic actinomycetes, whose average inhibition diameters were more than 20 mm. We identified 3 strains of actinomycetes according to culture characteristics on differential culture mediums, spore and spore chain morphology, physiological and biochemical, molecular biology methods. The results showed that the 3 strains of actinomycetes were Streptomyces roseofulvus, Streptomyces olivaceoviridis and Streptomyces corchorusii. Among them, the best was S. roseofulvus, and average inhibition diameter was 54.66 mm. Average inhibition diameter of S.olivaceoviridis was 43.20 mm, and average inhibition diameter of S. corchorusii was 20.34 mm.%  为筛选出烟草青枯菌的有效拮抗菌,本研究从烟草根围土壤中分离了97株放线菌,通过平板对峙培养法筛选出抑菌圈直径均达20 mm以上的拮抗放线菌3株。根据其在鉴别培养基上的培养特征、孢子和孢子链的形态特征,生理生化特性以及16SrDNA序列分析对这3株拮抗放线菌进行鉴定。结果表明,3株拮抗放线菌都属于链霉菌属(Streptomyces),分别为粉红孢类群中的玫瑰暗黄链霉菌(S. roseofulvus Preobrazhenskaya)、绿色类群中的橄榄绿链霉菌(S. olivaceoviridis Preobrazhenskaya)和灰褐类群中的黄麻链霉菌(S. corchorusii Ahmed)。其中,玫瑰暗黄链霉菌菌株对青枯劳尔氏菌的拮抗效果最好,其平均抑菌圈直径可达54.66 mm;橄榄绿链霉菌菌株的拮抗效果次之,其平均抑菌圈直径为43.20 mm;黄麻链霉菌平均抑菌圈直径为20.34 mm。

  16. Diversity of actinomycetes in Taiwan strait marine sediments%台湾海峡海洋沉积物放线菌的多样性

    Institute of Scientific and Technical Information of China (English)

    梁效伟; 陈名洪; 林如; 王海龙; 谢阳; 连云阳; 江红

    2012-01-01

    目的 探究台湾海峡海洋沉积物中放线菌的多样性及发现合成药物先导化合物的新菌源.方法 采用6种选择性培养基分离15份来自台湾海峡沉积物样品中含有的放线菌.挑选不同培养特征的放线菌进行初步分类鉴别、16S rRNA基因序列系统进化分析及基于PCR的烯二炔抗生素基因筛选.结果 共分离到497株放线菌,挑选的95株放线菌分别属于放线菌7个科,11个属.16S rRNA基因序列分析结果提示分离到的小单孢菌科菌种存在数个潜在新种,95株菌中有27%的菌株含有烯二炔抗生素核弹头的生物合成基因片段.结论 海洋环境蕴含丰富的放线菌资源,具有产生烯二炔类抗生素的潜能.%Objective To investigate the diversity of actinomycetes isolated from Taiwan strait marine sediments and isolate new actinomycetes for discovering compounds of pharmaceutical importance. Methods Six selective media were used to isolate actinomycetes from 15 sediment samples. Actinobacterial diversity in these sediments was investigated by phylogenetic analysis based on 16S rRNA gene sequences. To detect potential producer strains of enediyne antibiotics, PCR based screening strategy was used. Results A total of 497 strains of actinomycetes were isolated and 95 representative isolates were selected on the basis of their morphologies on different media. 16S rRNA gene sequences phylogenetic analysis showed that these strains belonged to seven families including eleven genera. Phylogenetic analyses also grouped many of the strains into clades distinct from alt known genera within Micromonosporaceae, indicating that they may be new genera. 27% of the above 95 strains were detected and found containing enediyne polyketide synthase (PK.S) gene. Conclusion The results confirm that marine sediments are rich source of rare actinomycetes and the actinomycetes from marine environment have the potential of producing enediyne family antibiotics.

  17. 盐碱土壤放线菌的研究概况%Research Advances of Actinomycetes in Saline-alkali Soil

    Institute of Scientific and Technical Information of China (English)

    胡磊; 景彩虹; 薄乐涛; 达文燕; 杨建文; 姚健; 牛世全

    2012-01-01

    盐碱土壤放线菌是极端微生物的重要组成部分,也是一类极具应用前景的微生物资源.讨论了盐碱土壤放线菌的分离问题,并对其分类的发展与现状进行了概述,同时还就筛选抗生素高产菌株作了介绍.%Actinomycetes existing in saline—alkali soil are important members of extreme environmental microorganisms with wide application prospect. The screening methods for isolating actinmycetes from saline-alkali soil was discussed; and the development and current situation of their identification were briefly summarized. Furthermore, the screening of high antibiotics producing actinomycetes was also introduced.

  18. Isolation of ~richoderma sp. and Actinomycetes from Camation tDianthus caryophyllus Soil and Evaluation in vitro of their Antagonic Activity against Fusarium oxysporum. f. sp. dianthi Aislamiento de Trichoderma sp. y actinomycetes a partir de suelos de clavel (Dianthus caryophyllus y evaluación de su capacidad antagónica in vitro sobre Fusarium oxysporum. f. sp. Dianthi

    Directory of Open Access Journals (Sweden)

    Márquez Marcela

    2002-08-01

    Full Text Available Uno de los problemas más limitantes en el cultivo de clavel en Colombia es el marchitamiento vascular causado por Fusarium oxysporum f. sp. dianthi (Foxd el método empleado actualmente para controlar y/o prevenir esta enfermedad es la aplicación de fungicidas, los cuáles no son tan efectivos como se espera y al emplearse en exceso causan daños al medio ambiente. Por lo tanto el uso de poblaciones microbianas nativas para controlar esta enfermedad se perfila corno una alternativa importante en los programas de erradicación de la enfermedad. Algunas especies de Trichoderma y de Actinomycetes, se han estudiado, por la capacidad de producir sustancias inhibitorias del crecimiento y/o la actividad de este fitopatógeno. En este estudio se aislaron diversas cepas de estos microorganismos controladores y se evaluó in vitro su actividad antagónica.
    Se aislaron seis cepas de Trichoderma y treinta de Actinomycetes a partir de la rizósfera de diferentes
    cultivos de clavel de la Sabana de Bogotá; la inhibición del crecimiento de Foxd fue evaluada in vitro por medio de la interacción hongo-hongo y actinomycete-hongo al medir el porcentaje de inhibición micelial (%MI y la formación de un halo de inhibición alrededor del crecimiento de Foxd. Los aislamientos de Trichoderma sp y Actinomycetes mostraron un %MI mayor al 50%. El aislamiento VI de Trichoderma sp (T-VI presentó un %MI del 89% mientras que el aislamiento VII de Actinomycetes (A-VII, identificado como Streptomyces
    sp alcanzó un %MI del 91%, un halo de inhibición mayor a 1cm. Posteriormente, fue imposible determinar la actividad antagónica en asociación entre los aislamientos T-VI y A-VII debido al efecto inhibitorio de Streptomyces sp sobre  Trichoderma sp.
    One of the major problems of the carnation crop in
    Colombia is the vascular wilt disease caused by Fusarium
    oxysporum f.. sp. dianthi (Foxd. Since the only method currently used to control andlor

  19. In vitro and in vivo antagonism of actinomycetes isolated from Moroccan rhizospherical soils against Sclerotium rolfsii: a causal agent of root rot on sugar beet (Beta vulgaris L.).

    Science.gov (United States)

    Errakhi, R; Lebrihi, A; Barakate, M

    2009-08-01

    To evaluate the ability of the isolated actinomycetes to inhibit in vitro plant pathogenic fungi and the efficacy of promising antagonistic isolates to reduce in vivo the incidence of root rot induced by Sclerotium rolfsii on sugar beet. Actinomycetes isolated from rhizosphere soil of sugar beet were screened for antagonistic activity against a number of plant pathogens, including S. rolfsii. Ten actinomycetes out of 195 screened in vitro were strongly inhibitory to S. rolfsii. These isolates were subsequently tested for their ability to inhibit sclerotial germination and hyphal growth of S. roflsii. The most important inhibitions were obtained by the culture filtrate from the isolates J-2 and B-11, including 100% inhibition of sclerotial germination and 80% inhibition of hyphal growth. These two isolates (J-2 and B-11) were then screened for their ability to protect sugar beet against infection of S. rolfsii induced root rot in a pot trial. The treatment of S. rolfsii infested soil with a biomass and culture filtrate mixture of the selected antagonists reduced significantly (P beet. Isolate J-2 was most effective and allowed a high fresh weight of sugar beet roots to be obtained. Both antagonists J-2 and B-11 were classified as belonging to the genus Streptomyces species through morphological and chemical characteristics as well as 16S rDNA analysis. Streptomyces isolates J-2 and B-11 showed a potential for controlling root rot on sugar beet and could be useful in integrated control against diverse soil borne plant pathogens. This investigation showed the role, which actinomycete bacteria can play to control root rot caused by S. rolfsii, in the objective to reduce treatments with chemical fungicides.

  20. Quantitative Use of Fluorescent In Situ Hybridization To Examine Relationships between Mycolic Acid-Containing Actinomycetes and Foaming in Activated Sludge Plants

    OpenAIRE

    Davenport, Russell J.; Curtis, Thomas P; GOODFELLOW, Michael; Stainsby, Fiona M.; Bingley, Marc

    2000-01-01

    The formation of viscous foams on aeration basins and secondary clarifiers of activated sludge plants is a common and widespread problem. Foam formation is often attributed to the presence of mycolic acid-containing actinomycetes (mycolata). In order to examine the relationship between the number of mycolata and foam, we developed a group-specific probe targeting the 16S rRNA of the mycolata, a protocol to permeabilize mycolata, and a statistically robust quantification method. Statistical an...

  1. Use of remazol blue dyed avicel for the determination of cellulolytic activity in basidiomycetes Uso de Avicel colorida com Remazol Blue para determinação da atividade celulolítica em Basidiomycetos

    OpenAIRE

    Marcos José Correia; José Antônio de Sousa Pereira Junior; Jefferson Cunha dos Santos; Maria Auxiliadora de Queiroz Cavalcanti

    1998-01-01

    A modified method for direct determination of cellulolytic activity using Avicel colored with Remazol Brilliant Blue R (RBBR) in Agar test tubes is discussed. Refinements were introduced in a simple method for quantitation of cellulase activity, based on the release of dye from Avicel-RBBR medium by the enzymatic hydrolysis. Modifications in Avicel-dye preparation were enhanced and a spectrophotometer for direct OD measurement in agar test tubes used. The use of a spectrophotometer improved t...

  2. Structure of an MmyB-like regulator from C. aurantiacus, member of a new transcription factor family linked to antibiotic metabolism in actinomycetes.

    Directory of Open Access Journals (Sweden)

    Qingping Xu

    Full Text Available Actinomycetes are important bacterial sources of antibiotics and other secondary metabolites. Many antibiotic gene clusters are controlled by pathway-specific activators that act in response to growth conditions. Here we present the crystal structure of an MmyB-like transcription regulator MltR (PDB code 3pxp (Caur_2278 from Chloroflexus aurantiacus, in complex with a fatty acid (myristic acid. MltR is a distant homolog of the methylenomycin activator MmyB and consists of an Xre-type N-terminal DNA-binding domain and a C-terminal ligand-binding module that is related to the Per-Arnt-Sim (PAS domain. This structure has enabled identification of a new family of bacterial transcription factors that are distributed predominantly in actinomycetes. Bioinformatics analysis of MltR and other characterized family members suggest that they are likely associated with antibiotic and fatty acid metabolism in actinomycetes. Streptomyces coelicolor SCO4944 is a candidate as an ancestral member of the family. Its ortholog in S. griseus, SGR_6891, is induced by A-factor, a γ-butyrolactone that controls antibiotic production and development, and is adjacent to the A-factor synthase gen, afsA. The location of mltR/mmyB homologs, in particular those adjacent to less well-studied antibiotic-related genes, makes them interesting genetic markers for identifying new antibiotic genes. A model for signal-triggered DNA-binding by MltR is proposed.

  3. The Bio - control and Application of Actinomycetes against Plant Diseases%放线菌对植物病害的防治作用及应用

    Institute of Scientific and Technical Information of China (English)

    贾雨; 贾丽苑; 黄建新

    2012-01-01

    Actinomycetes are one of the most important bio - control micro - organisms which produce antibiotics and enzymes and are advantageous in the bio - control of plant diseases. This study addresses the bio - control of actinomycetes against plant diseases in terms of actinomycetes' action on phytopathogen and on the plants' disease resistance under the soil environ- ment. The paper also gives a brief introduction to the applications of actinomycetes in the bio - control of plant diseases, research hotspot and development trend.%放线菌是产生抗生素和酶的重要微生物资源之一,在防治植物病害中有很多优势.主要从放线菌与植物病原菌的作用和土壤环境中的放线菌对植物的抗病作用两方面,介绍了放线菌对植物病害的防治作用;简述目前放线菌在植病生防中的应用状况;以及研究热点和发展趋势.

  4. Recent advances in the bioactive metabolites of marine actinomycetes%海洋放线菌活性代谢产物研究最新进展

    Institute of Scientific and Technical Information of China (English)

    朱峰; 刘晓红; 林永成

    2007-01-01

    近年来海洋放线菌代谢产物的研究取得了很大进展,从海洋放线菌中分离到许多结构新奇、有特殊生理活性和有潜在实用价值的新化合物.研究表明海洋放线菌有可能成为抗生紊等药物工业的又一重要微生物资源.本文分类介绍了2001年到2005年间海洋放线菌代谢产物的研究进展,重点在于从海洋放线菌发现的新化合物及其生物活性.%Studis on the metabolites of marine actinomycetes have been rapidly developed recently.A number of unique structural compounds with special bioactivities and potential values were isolated from marine actinomycetes,which were expected to be another important microorganism resources for pharmaceutical industries.The recent advances in the bioactive metabolites of marine actinomycetes were reviewed in this paper with the literature published during 2001~2005.The emphasis is on the novel compounds and the relative bioactivities based on the structural classification.

  5. Screening of Mangrove Forest Actinomycetes and Its Antitumor Activity Detection%红树林放线菌筛选及其抗肿瘤活性测定

    Institute of Scientific and Technical Information of China (English)

    周中流; 徐立军

    2012-01-01

    Objective To isolate and purify the microbial strains from sea mud samples collected in Zhanjiang mangrove wetland for screening and detecting the antitumor activity. Methods The morphological method was adopted to identify actinomycetes strains. The MTT assay was applied to measure two tumor cells lines ( A549 and K562 ) cytotoxicity of 72 strains of actinomycete fermentation broth. Results By identification of isolated 72 strains of actinomycete, 18 percents of actinomycete fermentation broth showed the cytotoxicity in varying degrees. Especially,the fermentation broth of N2010-37 and N2010-68 revealed obvious antitumor activity on the above - mentioned two tumor cells lines. Conclusion The research results establish the foundation for seeking the antitumor components from mangrove forest actinomycete in Zhanjiang.%目的 从我国湛江红树林采集的海泥样品中分离纯化微生物菌株并进行筛选及抗肿瘤活性测定.方法 采用形态学方法鉴定放线菌菌株;采用四氮唑盐(MTT)法测定筛选出的72株放线菌发酵液对肺癌细胞A549与人类慢性髓性白血病细胞K562两种肿瘤细胞的细胞毒活性.结果 经鉴定分离得到了72株放线菌,其中18%的放线菌发酵液具有不同程度的细胞毒活性.N2010-37和N2010-68两株放线菌发酵液对上述两种肿瘤细胞株作用较显著.结论 该研究结果为从湛江红树林放线菌中寻找抗肿瘤活性成分奠定了基础.

  6. Comparative study of different methods for isolation of marine actinomycetes%海洋放线菌不同分离方法的比较研究

    Institute of Scientific and Technical Information of China (English)

    常显波; 刘文正; 尹琦; 张晓华

    2012-01-01

    The isolation techniques for marine actinomycetes from the inter-tidal sediment at Qingdao were studied by using the series dilution and plate spreading methods. The impacts of different pretreatments, diluents, seawater concentrations and media on the isolation of actinomycetes were investigated. The results showed that the growth of bacteria were obviously restrained in the samples when pretreated with 55℃ for 6 minutes, which enhanced the isolation of actinomycetes from the inter-tidal sediment; dilution of the samples with 1/4 Ringer's solution and spreading them on the media prepared with pure seawater could improve the isolation of actinomycetes. The 9 media exhibited significant differences on the number of actinomycetes recovered, with media Ml, M6, M7 and M8 being more effective than others.%采用平板涂布法,以青岛海区潮间带沉积物为对象进行海洋放线菌的分离方法研究.具体分析了不同样品预处理方式、稀释液、海水浓度和培养基种类等对分离效果的影响.结果表明,55℃预处理样品6 min能有效减少细菌数量,利于潮间带沉积物中放线菌的分离;以1/4林格氏溶液稀释样品、纯海水配置培养基,可以分离得到较多的放线菌;不同培养基对沉积物中放线菌的分离效果差别很大,本实验设置的M1 ~ M9培养基中,M1、M6、M7和M8培养基的分离效果优于其他5种.

  7. Chapter 5. Applying the genetics of secondary metabolism in model actinomycetes to the discovery of new antibiotics.

    Science.gov (United States)

    van Wezel, Gilles P; McKenzie, Nancy L; Nodwell, Justin R

    2009-01-01

    The actinomycetes, including in particular members of the filamentous genus Streptomyces, are the industrial source of a large number of bioactive small molecules employed as antibiotics and other drugs. They produce these molecules as part of their "secondary" or nonessential metabolism. The number and diversity of secondary metabolic pathways is enormous, with some estimates suggesting that this one genus can produce more than 100,000 distinct molecules. However, the discovery of new antimicrobials is hampered by the fact that many wild isolates fail to express all or sometimes any of their secondary metabolites under laboratory conditions. Furthermore, the use of previously successful screening strategies frequently results in the rediscovery of known molecules: the all-important novel structures have proven to be elusive. Mounting evidence suggests that streptomycetes possess many regulatory pathways that control the biosynthetic gene clusters for these secondary metabolic pathways and that cell metabolism plays a significant role in limiting or potentiating expression as well. In this article we explore the idea that manipulating metabolic conditions and regulatory pathways can "awaken" silent gene clusters and lead to the discovery of novel antimicrobial activities.

  8. Chromomycins A2 and A3 from Marine Actinomycetes with TRAIL Resistance-Overcoming and Wnt Signal Inhibitory Activities

    Directory of Open Access Journals (Sweden)

    Kazufumi Toume

    2014-06-01

    Full Text Available A biological screening study of an actinomycetes strain assembly was conducted using a cell-based cytotoxicity assay. The CKK1019 strain was isolated from a sea sand sample. Cytotoxicity-guided fractionation of the CKK1019 strain culture broth, which exhibited cytotoxicity, led to the isolation of chromomycins A2 (1 and A3 (2. 1 and 2 showed potent cytotoxicity against the human gastric adenocarcinoma (AGS cell line (IC50 1; 1.7 and 2; 22.1 nM, as well as strong inhibitory effects against TCF/β-catenin transcription (IC50 1; 1.8 and 2; 15.9 nM. 2 showed the ability to overcome tumor necrosis factor (TNF-related apoptosis-inducing ligand (TRAIL resistance. To the best of our knowledge, the effects of chromomycins A2 (1 and A3 (2 on TRAIL resistance-overcoming activity, and on the Wnt signaling pathway, have not been reported previously. Thus, 1 and 2 warrant potential drug lead studies in relation to TRAIL-resistant and Wnt signal-related diseases and offer potentially useful chemical probes for investigating TRAIL resistance and the Wnt signaling pathway.

  9. Chromomycins A2 and A3 from marine actinomycetes with TRAIL resistance-overcoming and Wnt signal inhibitory activities.

    Science.gov (United States)

    Toume, Kazufumi; Tsukahara, Kentaro; Ito, Hanako; Arai, Midori A; Ishibashi, Masami

    2014-06-05

    A biological screening study of an actinomycetes strain assembly was conducted using a cell-based cytotoxicity assay. The CKK1019 strain was isolated from a sea sand sample. Cytotoxicity-guided fractionation of the CKK1019 strain culture broth, which exhibited cytotoxicity, led to the isolation of chromomycins A2 (1) and A3 (2). 1 and 2 showed potent cytotoxicity against the human gastric adenocarcinoma (AGS) cell line (IC50 1; 1.7 and 2; 22.1 nM), as well as strong inhibitory effects against TCF/β-catenin transcription (IC50 1; 1.8 and 2; 15.9 nM). 2 showed the ability to overcome tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) resistance. To the best of our knowledge, the effects of chromomycins A2 (1) and A3 (2) on TRAIL resistance-overcoming activity, and on the Wnt signaling pathway, have not been reported previously. Thus, 1 and 2 warrant potential drug lead studies in relation to TRAIL-resistant and Wnt signal-related diseases and offer potentially useful chemical probes for investigating TRAIL resistance and the Wnt signaling pathway.

  10. Anti-Allergic Compounds from the Deep-Sea-Derived Actinomycete Nesterenkonia flava MCCC 1K00610

    Directory of Open Access Journals (Sweden)

    Chun-Lan Xie

    2017-03-01

    Full Text Available A novel cyclic ether, nesterenkoniane (1, was isolated from the deep-sea-derived actinomycete Nesterenkonia flava MCCC 1K00610, together with 12 known compounds, including two macrolides (2, 3, two diketopiperazines (4, 5, two nucleosides (6, 7, two indoles (8, 9, three phenolics (10–12, and one butanol derivate (13. Their structures were established mainly on detailed analysis of the NMR and MS spectroscopic data. All 13 compounds were tested for anti-allergic activities using immunoglobulin E (IgE mediated rat mast RBL-2H3 cell model. Under the concentration of 20 μg/mL, 1 exhibited moderate anti-allergic activity with inhibition rate of 9.86%, compared to that of 37.41% of the positive control, loratadine. While cyclo(d-Pro-(d-Leu (4 and indol-3-carbaldehyde (8 showed the most potent effects with the IC50 values of 69.95 and 57.12 μg/mL, respectively, which was comparable to that of loratadine (IC50 = 35.01 μg/mL. To the best of our knowledge, it is the first report on secondary metabolites from the genus of Nesterenkonia.

  11. Pseudonocardians A–C, New Diazaanthraquinone Derivatives from a Deap-Sea Actinomycete Pseudonocardia sp. SCSIO 01299

    Directory of Open Access Journals (Sweden)

    Xianwen Yang

    2011-08-01

    Full Text Available Pseudonocardians A–C (2–4, three new diazaanthraquinone derivatives, along with a previously synthesized compound deoxynyboquinone (1, were produced by the strain SCSIO 01299, a marine actinomycete member of the genus Pseudonocardia, isolated from deep-sea sediment of the South China Sea. The structures of compounds 1–4 were determined by mass spectrometry and NMR experiments (1H, 13C, HSQC, and HMBC. The structure of compound 1, which was obtained for the first time from a natural source, was confirmed by X-ray analysis. Compounds 1–3 exhibited potent cytotoxic activities against three tumor cell lines of SF-268, MCF-7 and NCI-H460 with IC50 values between 0.01 and 0.21 μm, and also showed antibacterial activities on Staphylococcus aureus ATCC 29213, Enterococcus faecalis ATCC 29212 and Bacillus thuringensis SCSIO BT01, with MIC values of 1–4 μg mL−1.

  12. A novel alkaloid from marine-derived actinomycete Streptomyces xinghaiensis with broad-spectrum antibacterial and cytotoxic activities.

    Directory of Open Access Journals (Sweden)

    Wence Jiao

    Full Text Available Due to the increasing emergence of drug-resistant bacteria and tumor cell lines, novel antibiotics with antibacterial and cytotoxic activities are urgently needed. Marine actinobacteria are rich sources of novel antibiotics, and here we report the discovery of a novel alkaloid, xinghaiamine A, from a marine-derived actinomycete Streptomyces xinghaiensis NRRL B24674(T. Xinghaiamine A was purified from the fermentation broth, and its structure was elucidated based on extensive spectroscopic analysis, including 1D and 2D NMR spectrum as well as mass spectrometry. Xinghaiamine A was identified to be a novel alkaloid with highly symmetric structure on the basis of sulfoxide functional group, and sulfoxide containing compound has so far never been reported in microorganisms. Biological assays revealed that xinghaiamine A exhibited broad-spectrum antibacterial activities to both Gram-negative persistent hospital pathogens (e.g. Acinetobacter baumannii, Pseudomonas aeruginosa and Escherichia coli and Gram-positive ones, which include Staphylococcus aureus and Bacillus subtilis. In addition, xinghaiamine A also exhibited potent cytotoxic activity to human cancer cell lines of MCF-7 and U-937 with the IC50 of 0.6 and 0.5 µM, respectively.

  13. Bioaugmenting anaerobic digestion of biosolids with selected strains of Bacillus, Pseudomonas, and Actinomycetes species for increased methanogenesis and odor control.

    Science.gov (United States)

    Duran, Metin; Tepe, Nalan; Yurtsever, Deniz; Punzi, Vito L; Bruno, Charles; Mehta, Raj J

    2006-12-01

    The objective of this study was to evaluate the effects of bioaugmenting anaerobic biosolids digestion with a commercial product containing selected strains of bacteria from genera Bacillus, Pseudomonas, and Actinomycetes, along with ancillary organic compounds containing various micronutrients. Specifically, the effects of the bioaugment in terms of volatile solids destruction and generation and fate of odor-causing compounds during anaerobic digestion and during storage of the digested biosolids were studied. Two bench-scale anaerobic digesters receiving primary and secondary clarifier biosolids from various full-scale biological wastewater treatment plants were operated. One of the digesters received the bioaugment developed by Organica Biotech, while the other was operated as control. The bioaugmented digester generated 29% more net CH(4) during the 8 weeks of operation. In addition, the average residual propionic acid concentration in the bioaugmented digester was 54% of that in the control. The monitoring of two organic sulfide compounds, methyl mercaptan (CH(3)SH) and dimethyl sulfide (CH(3)SCH(3)), clearly demonstrated the beneficial effects of the bioaugmentation in terms of odor control. The biosolids digested in the bioaugmented digester generated a negligible amount of CH(3)SH during 10 days of post-digestion storage, while CH(3)SH concentration in the control reached nearly 300 ppm(v) during the same period. Similarly, peak CH(3)SCH(3) generated by stored biosolids from the bioaugmented digester was only 37% of that from the control.

  14. Nematicidal activity of fervenulin isolated from a nematicidal actinomycete, Streptomyces sp. CMU-MH021, on Meloidogyne incognita.

    Science.gov (United States)

    Ruanpanun, Pornthip; Laatsch, Hartmut; Tangchitsomkid, Nuchanart; Lumyong, Saisamorn

    2011-06-01

    An isolate of the actinomycete, Streptomyces sp. CMU-MH021 produced secondary metabolites that inhibited egg hatch and increased juvenile mortality of the root-knot nematode Meloidogyne incognita in vitro. 16S rDNA gene sequencing showed that the isolate sequence was 99% identical to Streptomyces roseoverticillatus. The culture filtrates form different culture media were tested for nematocidal activity. The maximal activity against M. incognita was obtained by using modified basal (MB) medium. The nematicidal assay-directed fractionation of the culture broth delivered fervenulin (1) and isocoumarin (2). Fervenulin, a low molecular weight compound, shows a broad range of biological activities. However, nematicidal activity of fervenulin was not previously reported. The nematicidal activity of fervenulin (1) was assessed using the broth microdilution technique. The lowest minimum inhibitory concentrations (MICs) of the compound against egg hatch of M. incognita was 30 μg/ml and juvenile mortality of M. incognita increasing was observed at 120 μg/ml. Moreover, at the concentration of 250 μg/ml fervenulin (1) showed killing effect on second-stage nematode juveniles of M. incognita up to 100% after incubation for 96 h. Isocoumarin (2), another bioactive compound produced by Streptomyces sp. CMU-MH021, showed weak nematicidal activity with M. incognita.

  15. Streptomyces kanasensis sp. nov., an Antiviral Glycoprotein Producing Actinomycete Isolated from Forest Soil Around Kanas Lake of China.

    Science.gov (United States)

    Han, Lirong; Zhang, Guoqiang; Miao, Guopeng; Zhang, Xing; Feng, Juntao

    2015-12-01

    A filamentous actinomycete, designated strain ZX01(T), was isolated from forest soil around Kanas Lake of China. A polyphasic taxonomic study was carried out to establish the status of strain ZX01(T). Chemical and morphological properties of the isolate were similar to those of species of the genus Streptomyces. Analysis of the almost complete 16S rRNA gene sequence placed strain ZX01(T) in the genus Streptomyces where it formed a distinct phyletic line with recognized species of this genus. The strain exhibited the highest sequence similarities to Streptomyces lavendofoliae NBRC 12882(T) (99.1%), S. luridus NBRC 12793(T) (99.0%), S. lavendulocolor NBRC 12881(T) (99.0%), S. gobitricini NBRC 15419(T) (99.0%), and S. roseolilacinus NBRC 12815(T) (98.9%). Low DNA-DNA relatedness values of 54.0, 50.0, 60.0, 66.7, and 50.4%, respectively, were found between strain ZX01(T) and corresponding strains above. A number of phenotypic properties also enabled the isolate to be differentiated from related species of the genus Streptomyces. Therefore, it is proposed that strain ZX01(T) should be classified as the type strain of a novel species in the genus Streptomyces, Streptomyces kanasensis sp. nov. The type strain is ZX01(T) (= CGMCC 4893(T) =JCM 30232(T)).

  16. Degradation of carbonyl sulfide by Actinomycetes and detection of clade D of β-class carbonic anhydrase.

    Science.gov (United States)

    Ogawa, Takahiro; Kato, Hiromi; Higashide, Mitsuru; Nishimiya, Mami; Katayama, Yoko

    2016-09-25

    Carbonyl sulfide (COS) is an atmospheric trace gas and one of the sources of stratospheric aerosol contributing to climate change. Although one of the major sinks of COS is soil, the distribution of COS degradation ability among bacteria remains unclear. Seventeen out of 20 named bacteria belonging to Actinomycetales had COS degradation activity at mole fractions of 30 parts per million by volume (ppmv) COS. Dietzia maris NBRC 15801(T) and Mycobacterium sp. THI405 had the activity comparable to a chemolithoautotroph Thiobacillus thioparus THI115 that degrade COS by COS hydrolase for energy production. Among 12 bacteria manifesting rapid degradation at 30 ppmv COS, Dietzia maris NBRC 15801(T) and Streptomyces ambofaciens NBRC 12836(T) degraded ambient COS (∼500 parts per trillion by volume). Geodermatophilus obscurus NBRC 13315(T) and Amycolatopsis orientalis NBRC 12806(T) increased COS concentrations. Moreover, six of eight COS degrading bacteria isolated from soils had partial nucleotide sequences similar to that of the gene encoding clade D of β-class carbonic anhydrase, which included COS hydrolase. These results indicate the potential importance of Actinomycetes in the role of soils as sinks of atmospheric COS.

  17. Genetic transformation of marine Actinomycete sp. Isolate M048 and expression of a recombinant plasmid carrying the apc gene

    Institute of Scientific and Technical Information of China (English)

    HOU Yanhua; LI Fuchao; QIN Song; WANG Quanfu

    2006-01-01

    Optimal conditions for protoplasts formation of marine Actinomycete sp. isolate M048 were described, dense and disperse mycelia were cultured in SGGP medium, 0.5% glycine, lysozyme exposure (2 mg/cm3, 37 ℃, 40 min), and the concentration of sucrose in protoplast buffer was 0.4 mol/dm3 for keeping the balance of osmotic pressure. Using PEG-mediated protoplasts transformation, the transformation frequency was 89 transformants per microgramme of pIJ702. Meanwhile, an effective transformation procedure was established based on intergeneric conjugation from E. coli ET12567 (pUZ8002) using shuttle vectors pPM801, pPM803 and a(ψ)C31-derived integration vector pIJ8600 containing oriT and attP fragments. Transformation frequencies were 5.30×10-4±0.26×10-4, 8.92×10-4±0.19×10-4 and 6.38×10-5±0.41×10-5, respectively. Further, the heterologous expression of the allophycocyanin gene (apc) in the strain M048 was used to demonstrate this transformation system. SDS-PAGE and Western blot analysis confirmed the expression of recombinant APC (rAPC).

  18. Identification and characterization of an anaerobic ethanol-producing cellulolytic bacterial consortium from Great Basin hot springs with agricultural residues and energy crops.

    Science.gov (United States)

    Zhao, Chao; Deng, Yunjin; Wang, Xingna; Li, Qiuzhe; Huang, Yifan; Liu, Bin

    2014-09-01

    In order to obtain the cellulolytic bacterial consortia, sediments from Great Basin hot springs (Nevada, USA) were sampled and enriched with cellulosic biomass as the sole carbon source. The bacterial composition of the resulting anaerobic ethanol-producing celluloytic bacterial consortium, named SV79, was analyzed. With methods of the full-length 16S rRNA librarybased analysis and denaturing gradient gel electrophoresis, 21 bacteria belonging to eight genera were detected from this consortium. Clones with closest relation to the genera Acetivibrio, Clostridium, Cellulosilyticum, Ruminococcus, and Sporomusa were predominant. The cellulase activities and ethanol productions of consortium SV79 using different agricultural residues (sugarcane bagasse and spent mushroom substrate) and energy crops (Spartina anglica, Miscanthus floridulus, and Pennisetum sinese Roxb) were studied. During cultivation, consortium SV79 produced the maximum filter paper activity (FPase, 9.41 U/ml), carboxymethylcellulase activity (CMCase, 6.35 U/ml), and xylanase activity (4.28 U/ml) with sugarcane bagasse, spent mushroom substrate, and S. anglica, respectively. The ethanol production using M. floridulus as substrate was up to 2.63 mM ethanol/g using gas chromatography analysis. It has high potential to be a new candidate for producing ethanol with cellulosic biomass under anoxic conditions in natural environments.

  19. Cloning of novel cellulases from cellulolytic fungi: heterologous expression of a family 5 glycoside hydrolase from Trametes versicolor in Pichia pastoris.

    Science.gov (United States)

    Salinas, Alejandro; Vega, Marcela; Lienqueo, María Elena; Garcia, Alejandro; Carmona, Rene; Salazar, Oriana

    2011-12-10

    Total cDNA isolated from cellulolytic fungi cultured in cellulose was examined for the presence of sequences encoding for endoglucanases. Novel sequences encoding for glycoside hydrolases (GHs) were identified in Fusarium oxysporum, Ganoderma applanatum and Trametes versicolor. The cDNA encoding for partial sequences of GH family 61 cellulases from F. oxysporum and G. applanatum shares 58 and 68% identity with endoglucanases from Glomerella graminicola and Laccaria bicolor, respectively. A new GH family 5 endoglucanase from T. versicolor was also identified. The cDNA encoding for the mature protein was completely sequenced. This enzyme shares 96% identity with Trametes hirsuta endoglucanase and 22% with Trichoderma reesei endoglucanase II (EGII). The enzyme, named TvEG, has N-terminal family 1 carbohydrate binding module (CBM1). The full length cDNA was cloned into the pPICZαB vector and expressed as an active, extracellular enzyme in the methylotrophic yeast Pichia pastoris. Preliminary studies suggest that T. versicolor could be useful for lignocellulose degradation. Copyright © 2011 Elsevier Inc. All rights reserved.

  20. Similar is not the same: differences in the function of the (hemi-)cellulolytic regulator XlnR (Xlr1/Xyr1) in filamentous fungi.

    Science.gov (United States)

    Klaubauf, Sylvia; Narang, Hari Mander; Post, Harm; Zhou, Miaomiao; Brunner, Kurt; Mach-Aigner, Astrid R; Mach, Robert L; Heck, Albert J R; Altelaar, A F Maarten; de Vries, Ronald P

    2014-11-01

    The transcriptional activator XlnR (Xlr1/Xyr1) is a major regulator in fungal xylan and cellulose degradation as well as in the utilization of d-xylose via the pentose catabolic pathway. XlnR homologs are commonly found in filamentous ascomycetes and often assumed to have the same function in different fungi. However, a comparison of the saprobe Aspergillus niger and the plant pathogen Magnaporthe oryzae showed different phenotypes for deletion strains of XlnR. In this study wild type and xlnR/xlr1/xyr1 mutants of five fungi were compared: Fusarium graminearum, M. oryzae, Trichoderma reesei, A. niger and Aspergillus nidulans. Growth profiling on relevant substrates and a detailed analysis of the secretome as well as extracellular enzyme activities demonstrated a common role of this regulator in activating genes encoding the main xylanolytic enzymes. However, large differences were found in the set of genes that is controlled by XlnR in the different species, resulting in the production of different extracellular enzyme spectra by these fungi. This comparison emphasizes the functional diversity of a fine-tuned (hemi-)cellulolytic regulatory system in filamentous fungi, which might be related to the adaptation of fungi to their specific biotopes. Data are available via ProteomeXchange with identifier PXD001190. Copyright © 2014 Elsevier Inc. All rights reserved.