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Sample records for ammonia lyase microcompartment

  1. Preliminary structural investigations of the Eut-L shell protein of the ethanolamine ammonia-lyase metabolosome of Escherichia coli

    International Nuclear Information System (INIS)

    Nikolakakis, Kiel; Ohtaki, Akashi; Newton, Keith; Chworos, Arkadiusz; Sagermann, Martin

    2009-01-01

    Preliminary X-ray analysis of crystals of the bacterial microcompartment shell protein Eut-L from Escherichia coli is reported. The ethanolamine ammonia-lyase microcompartment is composed of five different shell proteins that have been proposed to assemble into symmetrically shaped polyhedral particles of varying sizes. Here, preliminary X-ray analysis of crystals of the bacterial microcompartment shell protein Eut-L from Escherichia coli is reported. Cloning, overexpression and purification resulted in highly pure protein that crystallized readily under many different conditions. In all cases the protein forms thin hexagonal plate-shaped crystals belonging to space group P3 that are of unusually high stability against different solvent conditions. The crystals diffracted to a resolution of 2.0 Å using synchrotron radiation but proved to be radiation-sensitive. Preparations of heavy-atom-derivatized crystals for use in determining the three-dimensional structure are under way

  2. Phenylalanine ammonia-lyase (PAL) gene activity in response to ...

    African Journals Online (AJOL)

    Phenylalanine ammonia-lyase (PAL) catalyzes the biosynthesis of rosmarinic acid (RA), tyrosine and phenylalanine are the precursors of RA, while proline drives metabolite precursors toward Shikimate and phenylpropanoid pathway ending with the production of RA. The aim of this study was to investigate the PAL gene ...

  3. Phenylalanine ammonia-lyase (PAL) gene activity in response to ...

    African Journals Online (AJOL)

    Yomi

    2012-01-03

    Jan 3, 2012 ... Phenylalanine ammonia-lyase (PAL) catalyzes the biosynthesis of rosmarinic acid (RA), tyrosine and phenylalanine are the precursors of RA, while proline drives metabolite precursors toward Shikimate and phenylpropanoid pathway ending with the production of RA. The aim of this study was to.

  4. Priming ammonia lyases and aminomutases for industrial and therapeutic applications

    NARCIS (Netherlands)

    Heberling, Matthew M.; Wu, Bian; Bartsch, Sebastian; Janssen, Dick B.

    Ammonia lyases (AL) and aminomutases (AM) are emerging in green synthetic routes to chiral amines and an AL is being explored as an enzyme therapeutic for treating phenylketonuria and cancer. Although the restricted substrate range of the wild-type enzymes limits their widespread application, the

  5. Phenylalanine ammonia-lyase through evolution: A bioinformatic approach

    Directory of Open Access Journals (Sweden)

    Shiva Hemmati

    2015-03-01

    Full Text Available Phenylalanine ammonia-lyase (PAL is the first entry enzyme of the phenylpropanoid pathway that converts phenylalanine to cinnamic acid which is the precursor of various secondary metabolites. PAL is recently formulated for phenylketonuric patients in pegylated forms; therefore, screening a PAL with the highest affinity to the substrate is of a great importance. PAL exists in all higher plants and some fungi and few bacteria. Ancestors of land plants have been adopted by evolving metabolic pathways. A multi-gene family encodes PAL by gene duplication events in most plants. In this study, the taxonomic distribution and phylogeny of pal gene found in land plants, fungi and bacteria have been analyzed. It seems that the ancestor of plants acquired a pal gene via horizontal gene transfer in symbioses with bacteria and fungi. Gymnosperms have kept a diverse set of pal genes that arose from gene duplication events. In angiosperms, after the divergence of dicotyledons from monocots, pal genes were duplicated many times. The close paralogues of pal genes in some species indicate expansion of gene families after the divergence in plant pal gene evolution. Interestingly, some of the plant pals clustered by species in a way that pals within one species are more closely related to each other than to homologs in the other species which indicates this duplication event occurred more recently.

  6. Genomic Characterization of Phenylalanine Ammonia Lyase Gene in Buckwheat.

    Directory of Open Access Journals (Sweden)

    Karthikeyan Thiyagarajan

    Full Text Available Phenylalanine Ammonia Lyase (PAL gene which plays a key role in bio-synthesis of medicinally important compounds, Rutin/quercetin was sequence characterized for its efficient genomics application. These compounds possessing anti-diabetic and anti-cancer properties and are predominantly produced by Fagopyrum spp. In the present study, PAL gene was sequenced from three Fagopyrum spp. (F. tataricum, F. esculentum and F. dibotrys and showed the presence of three SNPs and four insertion/deletions at intra and inter specific level. Among them, the potential SNP (position 949th bp G>C with Parsimony Informative Site was selected and successfully utilised to individuate the zygosity/allelic variation of 16 F. tataricum varieties. Insertion mutations were identified in coding region, which resulted the change of a stretch of 39 amino acids on the putative protein. Our Study revealed that autogamous species (F. tataricum has lower frequency of observed SNPs as compared to allogamous species (F. dibotrys and F. esculentum. The identified SNPs in F. tataricum didn't result to amino acid change, while in other two species it caused both conservative and non-conservative variations. Consistent pattern of SNPs across the species revealed their phylogenetic importance. We found two groups of F. tataricum and one of them was closely related with F. dibotrys. Sequence characterization information of PAL gene reported in present investigation can be utilized in genetic improvement of buckwheat in reference to its medicinal value.

  7. Catalytic Mechanisms and Biocatalytic Applications of Aspartate and Methylaspartate Ammonia Lyases

    NARCIS (Netherlands)

    de Villiers, Marianne; Veetil, Vinod Puthan; Raj, Hans; de Villiers, Jandre; Poelarends, Gerrit J.

    2012-01-01

    Ammonia lyases catalyze the formation of alpha-beta-unsaturated bonds by the elimination of ammonia from their substrates. This conceptually straightforward reaction has been the emphasis of many studies, with the main focus on the catalytic mechanism of these enzymes and/or the use of these enzymes

  8. Engineering methylaspartate ammonia lyase for the asymmetric synthesis of unnatural amino acids

    NARCIS (Netherlands)

    Raj, Hans; Szymanski, Wiktor; de Villiers, Jandre; Rozeboom, Henriëtte J.; Puthan Veetil, Vinod; Reis, Carlos R.; Villiers, Marianne de; Dekker, Frank J.; Wildeman, Stefaan de; Quax, Wim J.; Thunnissen, Andy-Mark W.H.; Feringa, Ben L.; Janssen, Dick B.; Poelarends, Gerrit J.

    The redesign of enzymes to produce catalysts for a predefined transformation remains a major challenge in protein engineering. Here, we describe the structure-based engineering of methylaspartate ammonia lyase (which in nature catalyses the conversion of 3-methylaspartate to ammonia and

  9. Processes for the production of hydroxycinnamic acids using polypeptides having tyrosine ammonia lyase activity

    DEFF Research Database (Denmark)

    2016-01-01

    The present invention generally relates to the field of biotechnology as it applies to the production of hydroxycinnamic acids using polypeptides having tyrosine ammonia lyase activity. More particularly, the present invention pertains to polypeptides having tyrosine ammonia lyase activity and high...... substrate specificity towards tyrosine, which makes them particularly suitable in the production of p-coumaric acid and other hydroxycinnamic acids. The present invention thus provides processes for the production of p-coumaric acid and other hydroxycinnamic acids employing these polypeptides as well...

  10. Reduced phenylalanine ammonia-lyase and tyrosine ammonia-lyase activities and lignin synthesis in wheat grown under low pressure sodium lamps

    Science.gov (United States)

    Guerra, D.; Anderson, A. J.; Salisbury, F. B.

    1985-01-01

    Wheat (Triticum aestivum L. cv Fremont) grown in hydroponic culture under 24-hour continuous irradiation at 560 to 580 micromoles per square meter per second from either metalhalide (MH), high pressure sodium (HPS), or low pressure sodium (LPS) lamps reached maturity in 70 days. Grain yields were similar under all three lamps, although LPS-grown plants lodged at maturity. Phenylalanine ammonia-lyase (PAL) and a tyrosine ammonia lyase (TAL) with lesser activity were detected in all extracts of leaf, inflorescence, and stem. Ammonia-lyase activities increased with age of the plant, and plants grown under the LPS lamp displayed PAL and TAL activities lower than wheat cultured under MH and HPS radiation. Greenhouse solar-grown wheat had the highest PAL and TAL activities. Lignin content of LPS-grown wheat was also significantly reduced from that of plants grown under MH or HPS lamps or in the greenhouse, showing a correlation with the reduced PAL and TAL activities. Ratios of far red-absorbing phytochrome to total phytochrome were similar for all three lamps, but the data do not yet warrant a conclusion about specific wavelengths missing from the LPS lamps that might have induced PAL and TAL activities in plants under the other lamps.

  11. Crystal structure and characterization of a novel L-serine ammonia-lyase from Rhizomucor miehei

    Energy Technology Data Exchange (ETDEWEB)

    Qin, Zhen [College of Food Science and Nutritional Engineering, Beijing Advanced Innovation Center of Food Nutrition and Human Health, China Agricultural University, Beijing 100083 (China); Yan, Qiaojuan [College of Engineering, China Agricultural University, Beijing 100083 (China); Ma, Qingjun [Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071 (China); Jiang, Zhengqiang, E-mail: zhqjiang@cau.edu.cn [College of Food Science and Nutritional Engineering, Beijing Advanced Innovation Center of Food Nutrition and Human Health, China Agricultural University, Beijing 100083 (China)

    2015-10-23

    L-serine ammonia-lyase, as a member of the β-family of pyridoxal-5′-phosphate (PLP) dependent enzymes, catalyzes the conversion of L-serine (L-threonine) to pyruvate (α-ketobutyrate) and ammonia. The crystal structure of L-serine ammonia-lyase from Rhizomucor miehei (RmSDH) was solved at 1.76 Å resolution by X-ray diffraction method. The overall structure of RmSDH had the characteristic β-family PLP dependent enzyme fold. It consisted of two distinct domains, both of which show the typical open twisted α/β structure. A PLP cofactor was located in the crevice between the two domains, which was attached to Lys52 by a Schiff-base linkage. Unique residue substitutions (Gly78, Pro79, Ser146, Ser147 and Thr312) were discovered at the catalytic site of RmSDH by comparison of structures of RmSDH and other reported eukaryotic L-serine ammonia-lyases. Optimal pH and temperature of the purified RmSDH were 7.5 and 40 °C, respectively. It was stable in the pH range of 7.0–9.0 and at temperatures below 40 °C. This is the first crystal structure of a fungal L-serine ammonia-lyase. It will be useful to study the catalytic mechanism of β-elimination enzymes and will provide a basis for further enzyme engineering. - Highlights: • The crystal structure of a fungal L-serine ammonia-lyase (RmSDH) was solved. • Five unique residue substitutions are found at the catalytic site of RmSDH. • RmSDH was expressed in Pichia. pastoris and biochemically characterized. • RmSDH has potential application in splitting D/L-serine.

  12. Biocatalytic Enantioselective Synthesis of N-Substituted Aspartic Acids by Aspartate Ammonia Lyase

    NARCIS (Netherlands)

    Weiner, Barbara; Poelarends, Gerrit J.; Janssen, Dick B.; Feringa, Ben L.

    2008-01-01

    The gene encoding aspartate ammonia lyase (aspB) from Bacillus sp. YM55-1 has been cloned and overexpressed, and the recombinant enzyme containing a C-terminal His6 tag has been purified to homogeneity and subjected to kinetic characterization. Kinetic studies have shown that the His6 tag does not

  13. Kinetic Resolution and Stereoselective Synthesis of 3-Substituted Aspartic Acids by Using Engineered Methylaspartate Ammonia Lyases

    NARCIS (Netherlands)

    Raj, Hans; Szymanski, Wiktor; Villiers, Jandré de; Puthan Veetil, Vinod; Quax, Wim J.; Shimamoto, Keiko; Janssen, Dick B.; Feringa, Ben L.; Poelarends, Gerrit J.

    2013-01-01

    Kinetic resolution and asymmetric synthesis of various valuable 3-substituted aspartic acids, which were obtained in fair to good yields with diastereomeric ratio values of up to >98:2 and enantiomeric excess values of up to >99 %, by using engineered methylaspartate ammonia lyases are described.

  14. Alteration of the Diastereoselectivity of 3-Methylaspartate Ammonia Lyase by Using Structure-Based Mutagenesis

    NARCIS (Netherlands)

    Raj, Hans; Weiner, Barbara; Puthan Veetil, Vinod; Reis, Carlos R.; Quax, Wim J.; Janssen, Dick B.; Feringa, Ben L.; Poelarends, Gerrit J.

    2009-01-01

    3-Methylaspartate ammonia-lyase (MAL) catalyzes the reversible amination of mesaconate to give both (2S,3S)-3-methylaspartic acid and (2S,3R)-3-methylaspartic acid as products. The deamination mechanism of MAL is likely to involve general base catalysis, in which a catalytic base abstracts the C3

  15. Probing reversible chemistry in coenzyme B12-dependent ethanolamine ammonia lyase with kinetic isotope effects

    OpenAIRE

    Jones, Alex R; Rentergent, Julius; Scrutton, NS; Hay, S

    2015-01-01

    Coenzyme B12-dependent enzymes such as ethanolamine ammonia lyase have remarkable catalytic power and some unique properties that enable detailed analysis of the reaction chemistry and associated dynamics. By selectively deuterating the substrate (ethanolamine) and/or the ?-carbon of the 5?-deoxyadenosyl moiety of the intrinsic coenzyme B12, it was possible to experimentally probe both the forward and reverse hydrogen atom transfers between the 5?-deoxyadenosyl radical and substrate during si...

  16. Production of Diamino propionic acid ammonia lyase by a new strain of Salmonella typhimurium PU011

    Directory of Open Access Journals (Sweden)

    Shiva Kumar Vasanth V

    2002-03-01

    Full Text Available Abstract Background Seeds of the legume plant Lathyrus sativus, which is grown in arid and semi arid tropical regions, contain Diamino Propionic acid (DAP. DAP is a neurotoxin, which, when consumed, causes a disease called Lathyrism. Lathryrism may manifest as Neurolathyrism or Osteolathyrism, in which the nervous system, and bone formation respectively, are affected. DAP ammonia lyase is produced by a few microorganisms such as Salmonella typhi, Salmonella typhimurium and Pseudomonas, and is capable of detoxifying DAP. Results S. typhimurium PU011, a non-virulent bacterial strain isolated in our lab, was found to produce DAP ammonia lyase enzyme when grown in minimal medium containing DAP. There was a direct correlation between biomass yield and enzyme activity, until 16 h post inoculation in minimal medium containing DAP. Following ammonium sulphate precipitation and passing through Sephadex G100, CM-Sephadex and DEAE-Sephacel for crude enzyme extract preparation, about 68-fold enzyme purity was obtained. The purified enzyme gave maximum activity at pH 8.0 and was stable up to 45 degrees C. The Km value for the substrate was found to be 0.685mM, calculated from a Line Weaver Burk plot. Conclusion A new bacterial strain, S.typhimurium PU 011, which is capable of producing DAP ammonia lyase, was isolated.

  17. Ammonia lyases and aminomutases as biocatalysts for the synthesis of α-amino and β-amino acids.

    Science.gov (United States)

    Turner, Nicholas J

    2011-04-01

    Ammonia lyases catalyse the reversible addition of ammonia to cinnamic acid (1: R=H) and p-hydroxycinnamic (1: R=OH) to generate L-phenylalanine (2: R=H) and L-tyrosine (2: R=OH) respectively (Figure 1a). Both phenylalanine ammonia lyase (PAL) and tyrosine ammonia lyase (TAL) are widely distributed in plants, fungi and prokaryotes. Recently there has been interest in the use of these enzymes for the synthesis of a broader range of L-arylalanines. Aminomutases catalyse a related reaction, namely the interconversion of α-amino acids to β-amino acids (Figure 1b). In the case of L-phenylalanine, this reaction is catalysed by phenylalanine aminomutase (PAM) and proceeds stereospecifically via the intermediate cinnamic acid to generate β-Phe 3. Ammonia lyases and aminomutases are related in sequence and structure and share the same active site cofactor 4-methylideneimidazole-5-one (MIO). There is currently interest in the possibility of using these biocatalysts to prepare a wide range of enantiomerically pure l-configured α-amino and β-amino acids. Recent reviews have focused on the mechanism of these MIO containing enzymes. The aim of this review is to review recent progress in the application of ammonia lyase and aminomutase enzymes to prepare enantiomerically pure α-amino and β-amino acids. Copyright © 2010 Elsevier Ltd. All rights reserved.

  18. Site-directed mutagenesis, kinetic and inhibition studies of aspartate ammonia lyase from Bacillus sp YM55-1

    NARCIS (Netherlands)

    Veetil, Vinod Puthan; Raj, Hans; Quax, Wim J.; Janssen, Dick B.; Poelarends, Gerrit J.

    Aspartate ammonia lyases (also referred to as aspartases) catalyze the reversible deamination of l-aspartate to yield fumarate and ammonia. In the proposed mechanism for these enzymes, an active site base abstracts a proton from C3 of l-aspartate to form an enzyme-stabilized enediolate intermediate.

  19. Highly Active and Specific Tyrosine Ammonia-Lyases from Diverse Origins Enable Enhanced Production of Aromatic Compounds in Bacteria and Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Jendresen, Christian Bille; Stahlhut, Steen Gustav; Li, Mingji

    2015-01-01

    Phenylalanine and tyrosine ammonia-lyases form cinnamic acid and p-coumaric acid, which are precursors of a wide range of aromatic compounds of biotechnological interest. Lack of highly active and specific tyrosine ammonia-lyases has previously been a limitation in metabolic engineering approaches...

  20. Expression and properties of the highly alkalophilic phenylalanine ammonia-lyase of thermophilic Rubrobacter xylanophilus.

    Directory of Open Access Journals (Sweden)

    Klaudia Kovács

    Full Text Available The sequence of a phenylalanine ammonia-lyase (PAL; EC: 4.3.1.24 of the thermophilic and radiotolerant bacterium Rubrobacter xylanophilus (RxPAL was identified by screening the genomes of bacteria for members of the phenylalanine ammonia-lyase family. A synthetic gene encoding the RxPAL protein was cloned and overexpressed in Escherichia coli TOP 10 in a soluble form with an N-terminal His6-tag and the recombinant RxPAL protein was purified by Ni-NTA affinity chromatography. The activity assay of RxPAL with l-phenylalanine at various pH values exhibited a local maximum at pH 8.5 and a global maximum at pH 11.5. Circular dichroism (CD studies showed that RxPAL is associated with an extensive α-helical character (far UV CD and two distinctive near-UV CD peaks. These structural characteristics were well preserved up to pH 11.0. The extremely high pH optimum of RxPAL can be rationalized by a three-dimensional homology model indicating possible disulfide bridges, extensive salt-bridge formation and an excess of negative electrostatic potential on the surface. Due to these properties, RxPAL may be a candidate as biocatalyst in synthetic biotransformations leading to unnatural l- or d-amino acids or as therapeutic enzyme in treatment of phenylketonuria or leukemia.

  1. Possible regulatory role of phenylalanine ammonia-lyase in the production of anthocyanins in asparagus (Asparagus officinalis L)

    NARCIS (Netherlands)

    Flores, F.B.; Oosterhaven, J.; Martinez-Madrid, M.C.; Romojaro, F.

    2005-01-01

    The regulatory role of phenylalanine ammonia-lyase (PAL) in the light-induced accumulation of anthocyanins in the epidermis of asparagus spears has been analysed. A correlation between the stimulation of PAL activity and the rise in total anthocyanin content has been observed. Light radiation

  2. Cooperative functioning between phenylalanine ammonia lyase and isochorishmate synthase activities contributes to salicylic acid biosynthesis in soybean

    Science.gov (United States)

    Salicylic acid (SA), an essential regulator of plant defense, is derived from chorismate via either the phenylalanine ammonia lyase (PAL), or the isochorishmate synthase (ICS) catalyzed steps. The ICS pathway is thought to be the primary contributor of defense-related SA, at least in Arabidopsis. We...

  3. Phenylalanine ammonia lyase catalyzed synthesis of amino acids by an MIO-cofactor independent pathway.

    Science.gov (United States)

    Lovelock, Sarah L; Lloyd, Richard C; Turner, Nicholas J

    2014-04-25

    Phenylalanine ammonia lyases (PALs) belong to a family of 4-methylideneimidazole-5-one (MIO) cofactor dependent enzymes which are responsible for the conversion of L-phenylalanine into trans-cinnamic acid in eukaryotic and prokaryotic organisms. Under conditions of high ammonia concentration, this deamination reaction is reversible and hence there is considerable interest in the development of PALs as biocatalysts for the enantioselective synthesis of non-natural amino acids. Herein the discovery of a previously unobserved competing MIO-independent reaction pathway, which proceeds in a non-stereoselective manner and results in the generation of both L- and D-phenylalanine derivatives, is described. The mechanism of the MIO-independent pathway is explored through isotopic-labeling studies and mutagenesis of key active-site residues. The results obtained are consistent with amino acid deamination occurring by a stepwise E1 cB elimination mechanism. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Comparative structural and enzymatic studies on Salmonella typhimurium diaminopropionate ammonia lyase reveal its unique features.

    Science.gov (United States)

    Deka, G; Bisht, S; Savithri, H S; Murthy, M R N

    2018-05-01

    Cellular metabolism of amino acids is controlled by a large number of pyridoxal 5'-phosphate (PLP) dependent enzymes. Diaminopropionate ammonia lyase (DAPAL), a fold type II PLP-dependent enzyme, degrades both the D and L forms of diaminopropionic acid (DAP) to pyruvate and ammonia. Earlier studies on the Escherichia coli DAPAL (EcDAPAL) had suggested that a disulfide bond located close to the active site may be crucial for maintaining the geometry of the substrate entry channel and the active site. In order to obtain further insights into the catalytic properties of DAPAL, structural and functional studies on Salmonella typhimurium DAPAL (StDAPAL) were initiated. The three-dimensional X-ray crystal structure of StDAPAL was determined at 2.5 Å resolution. As expected, the polypeptide fold and dimeric organization of StDAPAL is similar to those of EcDAPAL. A phosphate group was located in the active site of StDAPAL and expulsion of this phosphate is probably essential to bring Asp125 to a conformation suitable for proton abstraction from the substrate (D-DAP). The unique disulfide bond of EcDAPAL was absent in StDAPAL, although the enzyme displayed comparable catalytic activity. Site directed mutagenesis of the cysteine residues involved in disulfide bond formation in EcDAPAL followed by functional and biophysical studies further confirmed that the disulfide bond is not necessary either for substrate binding or for catalysis. The activity of StDAPAL but not EcDAPAL was enhanced by monovalent cations suggesting subtle differences in the active site geometries of these two closely related enzymes. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. Probing reversible chemistry in coenzyme B12 -dependent ethanolamine ammonia lyase with kinetic isotope effects.

    Science.gov (United States)

    Jones, Alex R; Rentergent, Julius; Scrutton, Nigel S; Hay, Sam

    2015-06-08

    Coenzyme B12 -dependent enzymes such as ethanolamine ammonia lyase have remarkable catalytic power and some unique properties that enable detailed analysis of the reaction chemistry and associated dynamics. By selectively deuterating the substrate (ethanolamine) and/or the β-carbon of the 5'-deoxyadenosyl moiety of the intrinsic coenzyme B12 , it was possible to experimentally probe both the forward and reverse hydrogen atom transfers between the 5'-deoxyadenosyl radical and substrate during single-turnover stopped-flow measurements. These data are interpreted within the context of a kinetic model where the 5'-deoxyadenosyl radical intermediate may be quasi-stable and rearrangement of the substrate radical is essentially irreversible. Global fitting of these data allows estimation of the intrinsic rate constants associated with CoC homolysis and initial H-abstraction steps. In contrast to previous stopped-flow studies, the apparent kinetic isotope effects are found to be relatively small. © 2015 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

  6. Probing Reversible Chemistry in Coenzyme B12-Dependent Ethanolamine Ammonia Lyase with Kinetic Isotope Effects

    Science.gov (United States)

    Jones, Alex R; Rentergent, Julius; Scrutton, Nigel S; Hay, Sam

    2015-01-01

    Coenzyme B12-dependent enzymes such as ethanolamine ammonia lyase have remarkable catalytic power and some unique properties that enable detailed analysis of the reaction chemistry and associated dynamics. By selectively deuterating the substrate (ethanolamine) and/or the β-carbon of the 5′-deoxyadenosyl moiety of the intrinsic coenzyme B12, it was possible to experimentally probe both the forward and reverse hydrogen atom transfers between the 5′-deoxyadenosyl radical and substrate during single-turnover stopped-flow measurements. These data are interpreted within the context of a kinetic model where the 5′-deoxyadenosyl radical intermediate may be quasi-stable and rearrangement of the substrate radical is essentially irreversible. Global fitting of these data allows estimation of the intrinsic rate constants associated with CoC homolysis and initial H-abstraction steps. In contrast to previous stopped-flow studies, the apparent kinetic isotope effects are found to be relatively small. PMID:25950663

  7. Biochemical analysis of Centaurea depressa phenylalanine ammonia lyase (PAL) for biotechnological applications in phenylketonuria (PKU).

    Science.gov (United States)

    Babaoğlu Aydaş, Selcen; Şirin, Seda; Aslim, Belma

    2016-12-01

    Phenylketonuria (PKU) is the most common hereditary defect of phenylalanine hydroxylase (PAH) enzyme achieving the hydroxylation of phenylalanine (Phe). Phenylalanine ammonia lyase (PAL) converts Phe to a harmless metabolite, trans-cinnamic acid (TCA) in plants and PAL enzyme activity is fairly high in plants rich in flavonoids. The study aimed the biochemical analysis of PAL form Centaurea depressa BIEB. (Asteraceae) a flavonoid rich plant. This study may form the main frame of future research efforts for the development of a plant preparation aimed for oral intake in PKU patients in an attempt to enrich their diet by allowing them to ingest some food stuff containing Phe without being exposed to complications. PAL was partially purified from the leaves of C. depressa. Enzyme activity was determined in comparison with that of other herbs that reportedly have a high PAL activity. Enzyme optimization was achieved and the PAL protein was detected by western blotting. C. depressa PAL demonstrated high activity (34.9 ± 0.6 U/mg protein). The enzyme was purified by 1.92-fold, which resulted in an activity of 53.30 ± 0.2 U/mg protein. The high-performance liquid chromatography analyzes of the PAL activity both before and after purification were in agreement. Western blot of PAL exhibited a 70 kDa protein band. The optimum pH and temperature are pH 8.8 and 37 °C. The optimum activities under gastric and intestinal digestion conditions were observed at pH 4.0 and pH 8.0, respectively. PAL activity of C. depressa is high, and does not disappear under different environmental conditions. This enzyme could be used for the development of dietary foods and biotechnological products for patients with PKU.

  8. Formulation and PEGylation optimization of the therapeutic PEGylated phenylalanine ammonia lyase for the treatment of phenylketonuria.

    Science.gov (United States)

    Bell, Sean M; Wendt, Dan J; Zhang, Yanhong; Taylor, Timothy W; Long, Shinong; Tsuruda, Laurie; Zhao, Bin; Laipis, Phillip; Fitzpatrick, Paul A

    2017-01-01

    Phenylketonuria (PKU) is a genetic metabolic disease in which the decrease or loss of phenylalanine hydroxylase (PAH) activity results in elevated, neurotoxic levels of phenylalanine (Phe). Due to many obstacles, PAH enzyme replacement therapy is not currently an option. Treatment of PKU with an alternative enzyme, phenylalanine ammonia lyase (PAL), was first proposed in the 1970s. However, issues regarding immunogenicity, enzyme production and mode of delivery needed to be overcome. Through the evaluation of PAL enzymes from multiple species, three potential PAL enzymes from yeast and cyanobacteria were chosen for evaluation of their therapeutic potential. The addition of polyethylene glycol (PEG, MW = 20,000), at a particular ratio to modify the protein surface, attenuated immunogenicity in an animal model of PKU. All three PEGylated PAL candidates showed efficacy in a mouse model of PKU (BTBR Pahenu2) upon subcutaneous injection. However, only PEGylated Anabaena variabilis (Av) PAL-treated mice demonstrated sustained low Phe levels with weekly injection and was the only PAL evaluated that maintained full enzymatic activity upon PEGylation. A PEGylated recombinant double mutant version of AvPAL (Cys503Ser/Cys565Ser), rAvPAL-PEG, was selected for drug development based on its positive pharmacodynamic profile and favorable expression titers. PEGylation was shown to be critical for rAvPAL-PEG efficacy as under PEGylated rAvPAL had a lower pharmacodynamic effect. rAvPAL and rAvPAL-PEG had poor stability at 4°C. L-Phe and trans-cinnamate were identified as activity stabilizing excipients. rAvPAL-PEG is currently in Phase 3 clinical trials to assess efficacy in PKU patients.

  9. Genome-wide characterization of phenylalanine ammonia-lyase gene family in watermelon (Citrullus lanatus).

    Science.gov (United States)

    Dong, Chun-Juan; Shang, Qing-Mao

    2013-07-01

    Phenylalanine ammonia-lyase (PAL), the first enzyme in the phenylpropanoid pathway, plays a critical role in plant growth, development, and adaptation. PAL enzymes are encoded by a gene family in plants. Here, we report a genome-wide search for PAL genes in watermelon. A total of 12 PAL genes, designated ClPAL1-12, are identified . Nine are arranged in tandem in two duplication blocks located on chromosomes 4 and 7, and the other three ClPAL genes are distributed as single copies on chromosomes 2, 3, and 8. Both the cDNA and protein sequences of ClPALs share an overall high identity with each other. A phylogenetic analysis places 11 of the ClPALs into a separate cucurbit subclade, whereas ClPAL2, which belongs to neither monocots nor dicots, may serve as an ancestral PAL in plants. In the cucurbit subclade, seven ClPALs form homologous pairs with their counterparts from cucumber. Expression profiling reveals that 11 of the ClPAL genes are expressed and show preferential expression in the stems and male and female flowers. Six of the 12 ClPALs are moderately or strongly expressed in the fruits, particularly in the pulp, suggesting the potential roles of PAL in the development of fruit color and flavor. A promoter motif analysis of the ClPAL genes implies redundant but distinctive cis-regulatory structures for stress responsiveness. Finally, duplication events during the evolution and expansion of the ClPAL gene family are discussed, and the relationships between the ClPAL genes and their cucumber orthologs are estimated.

  10. Partial rescue of neuropathology in the murine model of PKU following administration of recombinant phenylalanine ammonia lyase (pegvaliase).

    Science.gov (United States)

    Goldfinger, Marc; Zeile, William L; Corado, Carley R; O'Neill, Charles A; Tsuruda, Laurie S; Laipis, Philip J; Cooper, Jonathan D

    2017-09-01

    Pegylated recombinant phenylalanine ammonia lyase (pegvaliase) is an enzyme substitution therapy being evaluated for the treatment of phenylketonuria (PKU). PKU is characterized by elevated plasma phenylalanine, which is thought to lead to a deficiency in monoamine neurotransmitters and ultimately, neurocognitive dysfunction. A natural history evaluation in a mouse model of PKU demonstrated a profound decrease in tyrosine hydroxylase (TH) immunoreactivity in several brain regions, beginning at 4weeks of age. Following treatment with pegvaliase, the number of TH positive neurons was increased in several brain regions compared to placebo treated ENU2 mice. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  11. The phenylalanine ammonia lyase (PAL) gene family shows a gymnosperm-specific lineage

    Science.gov (United States)

    2012-01-01

    Background Phenylalanine ammonia lyase (PAL) is a key enzyme of the phenylpropanoid pathway that catalyzes the deamination of phenylalanine to trans-cinnamic acid, a precursor for the lignin and flavonoid biosynthetic pathways. To date, PAL genes have been less extensively studied in gymnosperms than in angiosperms. Our interest in PAL genes stems from their potential role in the defense responses of Pinus taeda, especially with respect to lignification and production of low molecular weight phenolic compounds under various biotic and abiotic stimuli. In contrast to all angiosperms for which reference genome sequences are available, P. taeda has previously been characterized as having only a single PAL gene. Our objective was to re-evaluate this finding, assess the evolutionary history of PAL genes across major angiosperm and gymnosperm lineages, and characterize PAL gene expression patterns in Pinus taeda. Methods We compiled a large set of PAL genes from the largest transcript dataset available for P. taeda and other conifers. The transcript assemblies for P. taeda were validated through sequencing of PCR products amplified using gene-specific primers based on the putative PAL gene assemblies. Verified PAL gene sequences were aligned and a gene tree was estimated. The resulting gene tree was reconciled with a known species tree and the time points for gene duplication events were inferred relative to the divergence of major plant lineages. Results In contrast to angiosperms, gymnosperms have retained a diverse set of PAL genes distributed among three major clades that arose from gene duplication events predating the divergence of these two seed plant lineages. Whereas multiple PAL genes have been identified in sequenced angiosperm genomes, all characterized angiosperm PAL genes form a single clade in the gene PAL tree, suggesting they are derived from a single gene in an ancestral angiosperm genome. The five distinct PAL genes detected and verified in P. taeda

  12. Structural And Biochemical Characterization of the Therapeutic A. Variabilis Phenylalanine Ammonia Lyase

    Energy Technology Data Exchange (ETDEWEB)

    Wang, L.; Gamez, A.; Archer, H.; Abola, E.E.; Sarkissian, C.N.; Fitzpatrick, P.; Wendt, D.; Zhang, Y.; Vellard, M.; Bliesath, J.; Bell, S.; Lemont, J.; Scriver, C.R.; Stevens, R.C.

    2009-05-26

    We have recently observed promising success in a mouse model for treating the metabolic disorder phenylketonuria with phenylalanine ammonia lyase (PAL) from Rhodosporidium toruloides and Anabaena variabilis. Both molecules, however, required further optimization in order to overcome problems with protease susceptibility, thermal stability, and aggregation. Previously, we optimized PAL from R. toruloides, and in this case we reduced aggregation of the A. variabilis PAL by mutating two surface cysteine residues (C503 and C565) to serines. Additionally, we report the structural and biochemical characterization of the A. variabilis PAL C503S/C565S double mutant and carefully compare this molecule with the R. toruloides engineered PAL molecule. Unlike previously published PAL structures, significant electron density is observed for the two active-site loops in the A. variabilis C503S/C565S double mutant, yielding a complete view of the active site. Docking studies and N-hydroxysuccinimide-biotin binding studies support a proposed mechanism in which the amino group of the phenylalanine substrate is attacked directly by the 4-methylidene-imidazole-5-one prosthetic group. We propose a helix-to-loop conformational switch in the helices flanking the inner active-site loop that regulates accessibility of the active site. Differences in loop stability among PAL homologs may explain the observed variation in enzyme efficiency, despite the highly conserved structure of the active site. A. variabilis C503S/C565S PAL is shown to be both more thermally stable and more resistant to proteolytic cleavage than R. toruloides PAL. Additional increases in thermal stability and protease resistance upon ligand binding may be due to enhanced interactions among the residues of the active site, possibly locking the active-site structure in place and stabilizing the tetramer. Examination of the A. variabilis C503S/C565S PAL structure, combined with analysis of its physical properties, provides

  13. The requirement of oxygen for the induction of phenylalanine ammonia lyase in potatoes by light or gamma irradiation

    International Nuclear Information System (INIS)

    Shirsat, S.G.; Nair, P.M.

    1976-01-01

    The induction of phenylalanine ammonia lyase (PAL) either in excised potato parenchymatous tissue by light or excised bud tissue by gamma radiation was abolished completely by submersion of the tissue in water. However, this effect could be reversed by passing air or oxygen vigorously through the medium. In this case the recovery of the induction was only about 75% of that of the air-incubated sample. Similarly the incubation of the tissue in nitrogen atmosphere instead of air caused complete inhibition of PAL induction. Experiments with protein synthesis inhibitors like cycloheximide, puromycin, actinomycin D and cinnabarinic acid, indicated that the oxygen-dependent enhancement of PAL activity, when the tissues are suspended in water, was due to de novo synthesis of the enzyme. (author)

  14. Site-directed mutagenesis, kinetic and inhibition studies of aspartate ammonia lyase from Bacillus sp. YM55-1.

    Science.gov (United States)

    Puthan Veetil, Vinod; Raj, Hans; Quax, Wim J; Janssen, Dick B; Poelarends, Gerrit J

    2009-06-01

    Aspartate ammonia lyases (also referred to as aspartases) catalyze the reversible deamination of L-aspartate to yield fumarate and ammonia. In the proposed mechanism for these enzymes, an active site base abstracts a proton from C3 of L-aspartate to form an enzyme-stabilized enediolate intermediate. Ketonization of this intermediate eliminates ammonia and yields the product, fumarate. Although two crystal structures of aspartases have been determined, details of the catalytic mechanism have not yet been elucidated. In the present study, eight active site residues (Thr101, Ser140, Thr141, Asn142, Thr187, His188, Lys324 and Asn326) were mutated in the structurally characterized aspartase (AspB) from Bacillus sp. YM55-1. On the basis of a model of the complex in which L-aspartate was docked manually into the active site of AspB, the residues responsible for binding the amino group of L-aspartate were predicted to be Thr101, Asn142 and His188. This postulate is supported by the mutagenesis studies: mutations at these positions resulted in mutant enzymes with reduced activity and significant increases in the K(m) for L-aspartate. Studies of the pH dependence of the kinetic parameters of AspB revealed that a basic group with a pK(a) of approximately 7 and an acidic group with a pK(a) of approximately 10 are essential for catalysis. His188 does not play the typical role of active site base or acid because the H188A mutant retained significant activity and displayed an unchanged pH-rate profile compared to that of wild-type AspB. Mutation of Ser140 and Thr141 and kinetic analysis of the mutant enzymes revealed that these residues are most likely involved in substrate binding and in stabilizing the enediolate intermediate. Mutagenesis studies corroborate the essential role of Lys324 because all mutations at this position resulted in mutant enzymes that were completely inactive. The substrate-binding model and kinetic analysis of mutant enzymes suggest that Thr187 and Asn326

  15. Phenylalanine ammonia-lyase (pal) and peroxidase activity in brown rust infected tissues of pakistani wheat cultivars

    International Nuclear Information System (INIS)

    Riaz, A.; Tahir, M.I.

    2014-01-01

    Besides other factors resistance and susceptibility is the outcome of biochemical processes such as activities of defense-related enzymes. So in this study, Phenylalanine ammonia-lyase (PAL) and Peroxidase activity of resistant (Inqilab-91) and susceptible (Kirin-95) wheat cultivars were determined through spectrophotometer to address the biochemical aspect related to the disease after 8 hours, 24 hours, 48 hours and 72 hours of leaf rust inoculation. The results have shown that these enzymes were present in both the resistant and susceptible cultivars but the activity was more pronounced in the resistant one. The effect of PAL and peroxidase activity was also investigated among inoculated and uninoculated plants within the same cultivar. The activity of both PAL and peroxidase were more significant in inoculated ones. The results have shown that the after 72 hours of inoculation Inqilab-91 had more PAL activity i.e., 5.47 IU/ml/min than in Kirin-95 i.e., 2.08 IU/ml/min at 270 nm. While peroxidase activity in Inqilab-91 was 6.41 IU/ml/min and in Kirin-95, 3.66 IU/ml/min after 72 hours of inoculation, observed under 470 nm wavelength. Increase in one's activity increases the other enzyme's activity. The activity was more prominent after 72 hours of infection as pathogen had successfully established itself in the host plant tissue. The activities of these enzymes act as plants active defense mechanism against the attack of pathogen. (author)

  16. [The relationship between activity and gene expression of phenylalanine ammonia-lyase and peel pitting in 'Fengjie' navel orange fruits].

    Science.gov (United States)

    Li, Zheng-Guo; Gao, Xue; Fan, Jing; Yang, Ying-Wu; Li, Dao-Gao; Kanellis, Angelos K

    2006-06-01

    Citrus fruit is prone to develop peel pitting during development and storage, which greatly decreases its fresh market value because of the deterioration of the peel. In the present study, we have examined the effect of different temperatures (15 degrees C and 4 degrees C), waxing and mechanical damage on the changes in the activity of phenylalanine ammonia-lyase (PAL) and the incidence of peel pitting in 'Fengjie' navel orange (Citrus sinensis Osbeck) fruits. The expression levels of PAL2, PAL6 genes in the peel during the development of peel pitting have been investigated through semi-quantitative PCR method. The incidence of peel pitting was greatly enhanced by waxing and mechanical damage and was decreased in lower temperature storage (4 degrees C) (Fig.1). Waxing and mechanical damage might be the important factors inducing peel pitting and suitable low temperature could decrease the incidence of this disease. The PAL activity increased during the whole storage period in accordance with the development of this pitting (Fig.2). The expression levels of PAL2 and PAL6 genes in damaged peel were higher than those in healthy peel and the expression of PAL2 is much more higher than that of PAL6 (Figs.4 and 5). The results suggested that the enzyme activity of PAL, along with the expression of PAL2 gene is highly related to this peel pitting occurred on 'Fengjie' navel orange fruits.

  17. Reduction of L-phenylalanine in protein hydrolysates using L-phenylalanine ammonia-lyase from Rhodosporidium toruloides.

    Science.gov (United States)

    Castañeda, María Teresita; Adachi, Osao; Hours, Roque Alberto

    2015-10-01

    L-Phenylalanine ammonia-lyase (PAL, EC 4.3.1.25) from Rhodosporidium toruloides was utilized to remove L-phenylalanine (L-Phe) from different commercial protein hydrolysates. A casein acid hydrolysate (CAH, L-Phe ~2.28 %) was employed as a model substrate. t-Cinnamic acid resulting from deamination of L-Phe was extracted, analyzed at λ = 290 nm, and used for PAL activity determination. Optimum reaction conditions, optimized using successive Doehlert design, were 35 mg mL(-1) of CAH and 800 mU mL(-1) of PAL, while temperature and pH were 42 °C and 8.7, respectively. Reaction kinetics of PAL with CAH was determined under optimized conditions. Then, removal of L-Phe from CAH was tested. Results showed that more than 92 % of initial L-Phe was eliminated. Similar results were obtained with other protein hydrolysates. These findings demonstrate that PAL is a useful biocatalyst for L-Phe removal from protein hydrolysates, which can be evaluated as potential ingredients in foodstuffs for PKU patients.

  18. Biochemical Evaluation of Phenylalanine Ammonia Lyase from Endemic Plant Cyathobasis fruticulosa (Bunge Aellen. for the Dietary Treatment of Phenylketonuria

    Directory of Open Access Journals (Sweden)

    Seda Şirin

    2016-01-01

    Full Text Available Enzyme substitution therapy with the phenylalanine ammonia lyase (PAL is a new approach to the treatment of patients with phenylketonuria (PKU. This enzyme is responsible for the conversion of phenylalanine to trans-cinnamic acid. We assessed the PAL enzyme of the endemic plant Cyathobasis fruticulosa (Bunge Aellen. for its possible role in the dietary treatment of PKU. The enzyme was found to have a high activity of (64.9±0.1 U/mg, with the optimum pH, temperature and buffer (Tris–HCl and L-phenylalanine concentration levels of pH=8.8, 37 °C and 100 mM, respectively. Optimum enzyme activity was achieved at pH=4.0 and 7.5, corresponding to pH levels of gastric and intestinal juice, and NaCl concentration of 200 mM. The purifi cation of the enzyme by 1.87-fold yielded an activity of 98.6 U/mg. PAL activities determined by HPLC analyses before and after purification were similar. Two protein bands, one at 70 and the other at 23 kDa, were determined by Western blot analysis of the enzyme. This enzyme is a potential candidate for serial production of dietary food and biotechnological products.

  19. Peroxidase and phenylalanine ammonia-lyase activities, phenolic acid contents, and allelochemicals-inhibited root growth of soybean

    Directory of Open Access Journals (Sweden)

    VANESSA HERRIG

    2002-01-01

    Full Text Available The influence of the allelochemicals ferulic (FA and vanillic (VA acids on peroxidase (POD, EC 1.11.1.7 and phenylalanine ammonia-lyase (PAL, EC 4.3.1.5 activities and their relationships with phenolic acid (PhAs contents and root growth of soybean (Glycine max (L. Merr. were examined. Three-day-old seedlings were cultivated in nutrient solution containing FA or VA (0.1 to 1 mM for 48 h. Both compounds (at 0.5 and 1 mM decreased root length (RL, fresh weight (FW and dry weight (DW and increased PhAs contents. At 0.5 and 1 mM, FA increased soluble POD activity (18% and 47%, respectively and cell wall (CW-bound POD activity (61% and 34%, while VA increased soluble POD activity (33% and 17% but did not affect CW-bound POD activity. At 1 mM, FA increased (82% while VA reduced (32% PAL activities. The results are discussed on the basis of the role of these compounds on phenylpropanoid metabolism and root growth and suggest that the effects caused on POD and PAL activities are some of the many mechanisms by which allelochemicals influence plant growth

  20. Entropic origin of cobalt-carbon bond cleavage catalysis in adenosylcobalamin-dependent ethanolamine ammonia-lyase.

    Science.gov (United States)

    Wang, Miao; Warncke, Kurt

    2013-10-09

    Adenosylcobalamin-dependent enzymes accelerate the cleavage of the cobalt-carbon (Co-C) bond of the bound coenzyme by >10(10)-fold. The cleavage-generated 5'-deoxyadenosyl radical initiates the catalytic cycle by abstracting a hydrogen atom from substrate. Kinetic coupling of the Co-C bond cleavage and hydrogen-atom-transfer steps at ambient temperatures has interfered with past experimental attempts to directly address the factors that govern Co-C bond cleavage catalysis. Here, we use time-resolved, full-spectrum electron paramagnetic resonance spectroscopy, with temperature-step reaction initiation, starting from the enzyme-coenzyme-substrate ternary complex and (2)H-labeled substrate, to study radical pair generation in ethanolamine ammonia-lyase from Salmonella typhimurium at 234-248 K in a dimethylsulfoxide/water cryosolvent system. The monoexponential kinetics of formation of the (2)H- and (1)H-substituted substrate radicals are the same, indicating that Co-C bond cleavage rate-limits radical pair formation. Analysis of the kinetics by using a linear, three-state model allows extraction of the microscopic rate constant for Co-C bond cleavage. Eyring analysis reveals that the activation enthalpy for Co-C bond cleavage is 32 ± 1 kcal/mol, which is the same as for the cleavage reaction in solution. The origin of Co-C bond cleavage catalysis in the enzyme is, therefore, the large, favorable activation entropy of 61 ± 6 cal/(mol·K) (relative to 7 ± 1 cal/(mol·K) in solution). This represents a paradigm shift from traditional, enthalpy-based mechanisms that have been proposed for Co-C bond-breaking in B12 enzymes. The catalysis is proposed to arise from an increase in protein configurational entropy along the reaction coordinate.

  1. Single-dose, subcutaneous recombinant phenylalanine ammonia lyase conjugated with polyethylene glycol in adult patients with phenylketonuria: an open-label, multicentre, phase 1 dose-escalation trial.

    Science.gov (United States)

    Longo, Nicola; Harding, Cary O; Burton, Barbara K; Grange, Dorothy K; Vockley, Jerry; Wasserstein, Melissa; Rice, Gregory M; Dorenbaum, Alejandro; Neuenburg, Jutta K; Musson, Donald G; Gu, Zhonghua; Sile, Saba

    2014-07-05

    Phenylketonuria is an inherited disease caused by impaired activity of phenylalanine hydroxylase, the enzyme that converts phenylalanine to tyrosine, leading to accumulation of phenylalanine and subsequent neurocognitive dysfunction. Phenylalanine ammonia lyase is a prokaryotic enzyme that converts phenylalanine to ammonia and trans-cinnamic acid. We aimed to assess the safety, tolerability, pharmacokinetic characteristics, and efficacy of recombinant Anabaena variabilis phenylalanine ammonia lyase (produced in Escherichia coli) conjugated with polyethylene glycol (rAvPAL-PEG) in reducing phenylalanine concentrations in adult patients with phenylketonuria. In this open-label, phase 1, multicentre trial, single subcutaneous injections of rAvPAL-PEG were given in escalating doses (0·001, 0·003, 0·010, 0·030, and 0·100 mg/kg) to adults with phenylketonuria. Participants aged 18 years or older with blood phenylalanine concentrations of 600 μmol/L or higher were recruited from among patients attending metabolic disease clinics in the USA. The primary endpoints were safety and tolerability of rAvPAL-PEG. Secondary endpoints were the pharmacokinetic characteristics of the drug and its effect on concentrations of phenylalanine. Participants and investigators were not masked to assigned dose group. This study is registered with ClinicalTrials.gov, number NCT00925054. 25 participants were recruited from seven centres between May 6, 2008, and April 15, 2009, with five participants assigned to each escalating dose group. All participants were included in the safety population. The most frequently reported adverse events were injection-site reactions and dizziness, which were self-limited and without sequelae. Two participants had serious adverse reactions to intramuscular medroxyprogesterone acetate, a drug that contains polyethylene glycol as an excipient. Three of five participants given the highest dose of rAvPAL-PEG (0·100 mg/kg) developed a generalised skin rash

  2. Change in the Content of Salicylic Acid and in the Activities of Phenylalanine Ammonia-Lyase and Catalase in Wheat Seedling Roots Under the Effect of Azospirillum Lectins

    Directory of Open Access Journals (Sweden)

    Alen'kina S.A.

    2012-05-01

    Full Text Available We investigated the time course of changes in the endogenous content of salicylic acid, the ratio between the acid's free and bound forms, and changes in the activities of phenylalanine ammonia-lyase and catalase in wheat seedling roots under the effect of the lectins of two strains of the associative nitrogen-fixing bacterium Azospirillum: A. brasilense Sp7 and its mutant defective in lectin activity, A. brasilense Sp7.2.3. Differences in plant response to the action of the lectins from these two strains were established. On the basis of the obtained data, a model was proposed for lectin-assisted induction of resistance, according to which the lectin effect on the roots of seedlings results in accumulation of free salicylic acid, which inhibits catalase activity, ultimately leading to accumulation of hydrogen peroxide and to formation of induced resistance.

  3. Structure and characterization of a cDNA clone for phenylalanine ammonia-lyase from cut-injured roots of sweet potato

    Energy Technology Data Exchange (ETDEWEB)

    Tanaka, Yoshiyuki; Matsuoka, Makoto; Yamanoto, Naoki; Ohashi, Yuko; Kano-Murakami, Yuriko; Ozeki, Yoshihiro (National Institute of Agro-Environmental Sciences, Ibaraki (Japan) Univ. of Tokyo (Japan))

    1989-08-01

    A cDNA clone for phenylalanine ammonia-lyase (PAL) induced in wounded sweet potato (Ipomoea batatas Lam.) root was obtained by immunoscreening a cDNA library. The protein produced in Escherichia coli cells containing the plasmid pPAL02 was indistinguishable from sweet potato PAL as judged by Ouchterlony double diffusion assays. The M{sub r} of its subunit was 77,000. The cells converted ({sup 14}C)-L-phenylalanine into ({sup 14}C)-t-cinnamic acid and PAL activity was detected in the homogenate of the cells. The activity was dependent on the presence of the pPAL02 plasmid DNA. The nucleotide sequence of the cDNA contained a 2,121-base pair (bp) open-reading frame capable of coding for a polypeptide with 707 amino acids (M{sub r} 77,137), a 22-bp 5{prime}-noncoding region and a 207-bp 3{prime}-noncoding region. The results suggest that the insert DNA fully encoded the amino acid sequence for sweet potato PAL that is induced by wounding. Comparison of the deduced amino acid sequence with that of a PAL cDNA fragment from Phaseolus vulgaris revealed 78.9% homology. The sequence from amino acid residues 258 to 494 was highly conserved, showing 90.7% homology.

  4. Structure and characterization of a cDNA clone for phenylalanine ammonia-lyase from cut-injured roots of sweet potato

    International Nuclear Information System (INIS)

    Tanaka, Yoshiyuki; Matsuoka, Makoto; Yamanoto, Naoki; Ohashi, Yuko; Kano-Murakami, Yuriko; Ozeki, Yoshihiro

    1989-01-01

    A cDNA clone for phenylalanine ammonia-lyase (PAL) induced in wounded sweet potato (Ipomoea batatas Lam.) root was obtained by immunoscreening a cDNA library. The protein produced in Escherichia coli cells containing the plasmid pPAL02 was indistinguishable from sweet potato PAL as judged by Ouchterlony double diffusion assays. The M r of its subunit was 77,000. The cells converted [ 14 C]-L-phenylalanine into [ 14 C]-t-cinnamic acid and PAL activity was detected in the homogenate of the cells. The activity was dependent on the presence of the pPAL02 plasmid DNA. The nucleotide sequence of the cDNA contained a 2,121-base pair (bp) open-reading frame capable of coding for a polypeptide with 707 amino acids (M r 77,137), a 22-bp 5'-noncoding region and a 207-bp 3'-noncoding region. The results suggest that the insert DNA fully encoded the amino acid sequence for sweet potato PAL that is induced by wounding. Comparison of the deduced amino acid sequence with that of a PAL cDNA fragment from Phaseolus vulgaris revealed 78.9% homology. The sequence from amino acid residues 258 to 494 was highly conserved, showing 90.7% homology

  5. DNA Methylation Influences Chlorogenic Acid Biosynthesis in Lonicera japonica by Mediating LjbZIP8 to Regulate Phenylalanine Ammonia-Lyase 2 Expression

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    Liangping Zha

    2017-07-01

    Full Text Available The content of active compounds differ in buds and flowers of Lonicera japonica (FLJ and L. japonica var. chinensis (rFLJ. Chlorogenic acid (CGAs were major active compounds of L. japonica and regarded as measurements for quality evaluation. However, little is known concerning the formation of active compounds at the molecular level. We quantified the major CGAs in FLJ and rFLJ, and found the concentrations of CGAs were higher in the buds of rFLJ than those of FLJ. Further analysis of CpG methylation of CGAs biosynthesis genes showed differences between FLJ and rFLJ in the 5′-UTR of phenylalanine ammonia-lyase 2 (PAL2. We identified 11 LjbZIP proteins and 24 rLjbZIP proteins with conserved basic leucine zipper domains, subcellular localization, and electrophoretic mobility shift assay showed that the transcription factor LjbZIP8 is a nuclear-localized protein that specifically binds to the G-box element of the LjPAL2 5′-UTR. Additionally, a transactivation assay and LjbZIP8 overexpression in transgenic tobacco indicated that LjbZIP8 could function as a repressor of transcription. Finally, treatment with 5-azacytidine decreased the transcription level of LjPAL2 and CGAs content in FLJ leaves. These results raise the possibility that DNA methylation might influence the recruitment of LjbZIP8, regulating PAL2 expression level and CGAs content in L. japonica.

  6. Characterization of the tissue-specific expression of phenylalanine ammonia-lyase gene promoter from loblolly pine (Pinus taeda) in Nicotiana tabacum.

    Science.gov (United States)

    Osakabe, Yuriko; Osakabe, Keishi; Chiang, Vincent L

    2009-09-01

    We isolated the 5' flanking region of a gene for phenylalanine ammonia-lyase (PAL; EC 4.3.1.5) from Pinus taeda, PtaPAL. To investigate the tissue-specific expression of the PtaPAL promoter, histochemical assay of GUS activity was performed using the transgenic tobacco expressing the PtaPAL promoter-GUS. The region of -897 to -420 in PtaPAL promoter showed high activities in the secondary xylem and response to bending stress. To characterize the cis-regulatory functions of the promoters for enzymes in phenylpropanoid biosynthesis, we examined the activity of chimeric promoters of PtaPAL and a 4-coumarate CoA ligase, Pta4CL alpha. The chimeric promoter showed similar activity as the Pta4CL alpha promoter. Electrophoretic mobility shift assays implicated -897 to -674 of PtaPAL promoter containing cis-elements of the expression in xylem of Pinus taeda. The results suggested that AC elements of PtaPAL have multiple functions in the expression under the various developmental stages and stress conditions in the transgenic tobacco.

  7. Expression and enzymatic activity of phenylalanine ammonia-lyase and p-coumarate 3-hydroxylase in mango (Mangifera indica 'Ataulfo') during ripening.

    Science.gov (United States)

    Palafox-Carlos, H; Contreras-Vergara, C A; Muhlia-Almazán, A; Islas-Osuna, M A; González-Aguilar, G A

    2014-05-16

    Phenylalanine ammonia lyase (PAL) and p-coumarate 3-hydroxylase (C3H) are key enzymes in the phenylpropanoid pathway. The relative expression of PAL and C3H was evaluated in mango fruit cultivar 'Ataulfo' in four ripening stages (RS1, RS2, RS3, and RS4) by quantitative polymerase chain reaction. In addition, enzyme activity of PAL and C3H was determined in mango fruits during ripening. The PAL levels were downregulated at the RS2 and RS3 stages, while C3H levels were upregulated in fruits only at RS3. The enzyme activity of PAL followed a pattern that was different from that of the PAL expression, thus suggesting regulation at several levels. For C3H, a regulation at the transcriptional level is suggested because a similar pattern was revealed by its activity and transcript level. In this study, the complexity of secondary metabolite biosynthesis regulation is emphasized because PAL and C3H enzymes are involved in the biosynthesis of several secondary metabolites that are active during all mango ripening stages.

  8. Isolation and Functional Characterization of a Phenylalanine Ammonia-Lyase Gene (SsPAL1 from Coleus (Solenostemon scutellarioides (L. Codd

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    Qinlong Zhu

    2015-09-01

    Full Text Available Phenylalanine ammonia-lyase (PAL is the first enzyme involved in the phenylpropanoid pathway and plays important roles in the secondary metabolisms, development and defense of plants. To study the molecular function of PAL in anthocyanin synthesis of Coleus (Solenostemon scutellarioides (L. Codd, a Coleus PAL gene designated as SsPAL1 was cloned and characterized using a degenerate oligonucleotide primer PCR and RACE method. The full-length SsPAL1 was 2450 bp in size and consisted of one intron and two exons encoding a polypeptide of 711 amino acids. The deduced SsPAL1 protein showed high identities and structural similarities with other functional plant PAL proteins. A series of putative cis-acting elements involved in transcriptional regulation, light and stress responsiveness were found in the upstream regulatory sequence of SsPAL1. Transcription pattern analysis indicated that SsPAL1 was constitutively expressed in all tissues examined and was enhanced by light and different abiotic factors. The recombinant SsPAL1 protein exhibited high PAL activity, at optimal conditions of 60 °C and pH 8.2. Although the levels of total PAL activity and total anthocyanin concentration have a similar variation trend in different Coleus cultivars, there was no significant correlation between them (r = 0.7529, p > 0.1, suggesting that PAL was not the rate-limiting enzyme for the downstream anthocyanin biosynthetic branch in Coleus. This study enables us to further understand the role of SsPAL1 in the phenylpropanoid (flavonoids, anthocyanins biosynthesis in Coleus at the molecular level.

  9. The Carboxysome and Other Bacterial Microcompartments

    Energy Technology Data Exchange (ETDEWEB)

    Kerfeld, Cheryl A.; Greenleaf, William B.; Kinney, James N.

    2010-06-23

    - Carboxysomes are part of the carbon concentrating mechanism in cyanobacteria and chemoautotrophs. - Carboxysomes are a subclass of bacterial microcompartments (BMCs); BMCs can encapsulate a range of metabolic processes. - Like some viral particles, the carboxysome can be modeled as an icosahedron-in its case, having 4,000-5,000 hexameric shell subunits and 12 surface pentamers to generate curvature. - The threefold axis of symmetry of the CsoS1D protein in carboxysomes forms a pore that can open and close, allowing for selective diffusion. - Genetic modules encoding BMC shell proteins and the enzymes that they encapsulate are horizontally transferable, suggesting they enable bacteria to adapt to diverse environments.

  10. Critical role of arginine 160 of the EutB protein subunit for active site structure and radical catalysis in coenzyme B12-dependent ethanolamine ammonia-lyase.

    Science.gov (United States)

    Sun, Li; Groover, Olivia A; Canfield, Jeffrey M; Warncke, Kurt

    2008-05-20

    The protein chemical, kinetic, and electron paramagnetic resonance (EPR) and electron spin-echo envelope modulation (ESEEM) spectroscopic properties of ethanolamine ammonia-lyase (EAL) from Salmonella typhimurium with site-directed mutations in a conserved arginine residue (R160) of the active site containing EutB protein subunit have been characterized. R160 was predicted by a comparative model of EutB to play a critical role in protein structure and catalysis [Sun, L., and Warncke, K. (2006) Proteins: Struct., Funct., Bioinf. 64, 308-319]. R160I and R160E mutants fail to assemble into an EAL oligomer that can be isolated by the standard enzyme purification procedure. The R160K and R160A mutants assemble, but R160A EAL is catalytically inactive and reacts with substrates to form magnetically isolated Co(II) and unidentified radical species. R160A EAL activity is resurrected by externally added guanidinium to 2.3% of wild-type EAL. R160K EAL displays catalytic turnover of aminoethanol, with a 180-fold lower value of k(cat)/ K(M) relative to wild-type enzyme. R160K EAL also forms Co(II)-substrate radical pair intermediate states during turnover on aminoethanol and (S)-2-aminopropanol substrates. Simulations of the X-band EPR spectra show that the Co(II)-substrate radical pair separation distances are increased by 2.1 +/- 1.0 A in R160K EAL relative to wild-type EAL, which corresponds to the predicted 1.6 A change in arginine versus lysine side chain length. 14N ESEEM from a hyperfine-coupled protein nitrogen in wild type is absent in R160K EAL, which indicates that a guanidinium 14N of R160 interacts directly with the substrate radical through a hydrogen bond. ESEEM of the 2H-labeled substrate radical states in wild-type and R160K EAL shows that the native separation distances among the substrate C1 and C2, and coenzyme C5' reactant centers, are conserved in the mutant protein. The EPR and ESEEM measurements evince a protein-mediated force on the C5'-methyl center

  11. Phenolics and Flavonoids Compounds, Phenylanine Ammonia Lyase and Antioxidant Activity Responses to Elevated CO2 in Labisia pumila (Myrisinaceae

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    Hawa Z.E. Jaafar

    2012-05-01

    Full Text Available A split plot 3 × 3 experiment was designed to examine the impact of three concentrations of CO2 (400, 800 and 1,200 µmol·mol−1 on the phenolic and flavonoid compound profiles, phenylalanine ammonia lyase (PAL and antioxidant activity in three varieties of Labisia pumila Benth. (var. alata, pumila and lanceolata after 15 weeks of exposure. HPLC analysis revealed a strong influence of increased CO2 concentration on the modification of phenolic and flavonoid profiles, whose intensity depended on the interaction between CO2 levels and L. pumila varieties. Gallic acid and quercetin were the most abundant phenolics and flavonoids commonly present in all the varieties. With elevated CO2 (1,200 µmol·mol−1 exposure, gallic acid increased tremendously, especially in var. alata and pumila (101–111%, whilst a large quercetin increase was noted in var. lanceolata (260%, followed closely by alata (201%. Kaempferol, although detected under ambient CO2 conditions, was undetected in all varieties after exposure. Instead, caffeic acid was enhanced tremendously in var. alata (338~1,100% and pumila (298~433%. Meanwhile, pyragallol and rutin were only seen in var. alata (810 µg·g−1 DW and pumila (25 µg·g−1 DW, respectively, under ambient conditions; but the former compound went undetected in all varieties while rutin continued to increase by 262% after CO2 enrichment. Interestingly, naringenin that was present in all varieties under ambient conditions went undetected under enrichment, except for var. pumila where it was enhanced by 1,100%. PAL activity, DPPH and FRAP also increased with increasing CO2 levels implying the possible improvement of health-promoting quality of Malaysian L. pumila

  12. Engineering self-assembled bioreactors from protein microcompartments

    Energy Technology Data Exchange (ETDEWEB)

    Savage, David [Univ. of California, Berkeley, CA (United States)

    2016-10-12

    The goals of this research are to understand how organisms such as bacteria segregate certain metabolic processes inside of specific structures, or “microcompartments,” in the cell and apply this knowledge to develop novel engineered microcompartments for use in nanotechnology and metabolic engineering. For example, in some bacteria, self-assembling protein microcompartments called carboxysomes encapsulate the enzymes involved in carbon fixation, enabling the cell to utilize carbon dioxide more effectively than if the enzymes were free in the cell. The proposed research will determine how structures such as carboxysomes assemble and function in bacteria and develop a means for creating novel, synthetic microcompartments for optimizing production of specific energy-rich compounds.

  13. Structural insight into the Clostridium difficile ethanolamine utilisation microcompartment.

    Directory of Open Access Journals (Sweden)

    Alison C Pitts

    Full Text Available Bacterial microcompartments form a protective proteinaceous barrier around metabolic enzymes that process unstable or toxic chemical intermediates. The genome of the virulent, multidrug-resistant Clostridium difficile 630 strain contains an operon, eut, encoding a bacterial microcompartment with genes for the breakdown of ethanolamine and its utilisation as a source of reduced nitrogen and carbon. The C. difficile eut operon displays regulatory genetic elements and protein encoding regions in common with homologous loci found in the genomes of other bacteria, including the enteric pathogens Salmonella enterica and Enterococcus faecalis. The crystal structures of two microcompartment shell proteins, CD1908 and CD1918, and an uncharacterised protein with potential enzymatic activity, CD1925, were determined by X-ray crystallography. CD1908 and CD1918 display the same protein fold, though the order of secondary structure elements is permuted in CD1908 and this protein displays an N-terminal β-strand extension. These proteins form hexamers with molecules related by crystallographic and non-crystallographic symmetry. The structure of CD1925 has a cupin β-barrel fold and a putative active site that is distinct from the metal-ion dependent catalytic cupins. Thin-section transmission electron microscopy of Escherichia coli over-expressing eut proteins indicates that CD1918 is capable of self-association into arrays, suggesting an organisational role for CD1918 in the formation of this microcompartment. The work presented provides the basis for further study of the architecture and function of the C. difficile eut microcompartment, its role in metabolism and the wider consequences of intestinal colonisation and virulence in this pathogen.

  14. Wound-induced ethylene synthesis and expression and formation of 1-aminocyclopropane-1-carboxylate (ACC) synthase, ACC oxidase, phenylalanine ammonia-lyase, and peroxidase in wounded mesocarp tissue of Cucurbita maxima.

    Science.gov (United States)

    Kato, M; Hayakawa, Y; Hyodo, H; Ikoma, Y; Yano, M

    2000-04-01

    1-Aminocyclopropane-1-carboxylate (ACC) synthase was rapidly induced in mesocarp tissue of Cucurbita maxima after wounding in the cut surface layer in 1 mm thickness (ca. 9 cells) (first layer) in both the enzyme activity and the levels of transcript. This led to a rapid accumulation of ACC and hence ethylene production. In the inside tissue (1-2 mm) (second layer), no significant induction of ACC synthase was observed, which resulted in a low level of ACC, although ethylene was evolved at a much lower rate than the first one. In contrast to ACC synthase, ACC oxidase was induced markedly in both the first and second layers and the development of its activity and the levels of mRNA remained high until later stages. It was considered that wound ethylene was closely associated with the development of ACC oxidase, since 2,5-norbornadiene (NBD), an inhibitor of ethylene action, substantially suppressed it. Phenylalanine ammonia-lyase (PAL) greatly increased in activity after wounding similarly to that of ACC synthase, in which increase in PAL activity occurred predominantly in the first layer. Induction of peroxidase activity after wounding had a close correlation in profile with that of ACC oxidase in that marked increases in the activity were observed in both the first and second layers and were strongly suppressed by NBD application. Four peroxidase isozymes were found by PAGE, among which a fraction was newly detected after wounding.

  15. Hydrogen Peroxide Treatment and the Phenylpropanoid Pathway Precursors Feeding Improve Phenolics and Antioxidant Capacity of Quinoa Sprouts via an Induction of L-Tyrosine and L-Phenylalanine Ammonia-Lyases Activities

    Directory of Open Access Journals (Sweden)

    Michał Świeca

    2016-01-01

    Full Text Available Hydrogen peroxide treatment and the phenylpropanoid pathway precursors feeding affected the antioxidant capacity of quinoa sprouts. Compared to the control, total phenolics content was significantly increased by treatment of control sprouts with 50 mM and 200 mM H2O2—an elevation of about 24% and 28%, respectively. The highest increase of flavonoids content was found for the sprouts treated with 200 mM H2O2 obtained from seeds fed with shikimic acid. All the studied modifications increased the antioxidant potential of sprouts (at least by 50% compared to control. The highest reducing power was found for the sprouts treated with 200 mM H2O2 obtained by phenylalanine feeding (5.03 mg TE/g DW and those obtained from the seeds fed with tyrosine (5.26 mg TE/g DW. The activities of L-tyrosine (TAL and L-phenylalanine (PAL ammonia-lyases were strongly affected by germination time as well as the applied modification of sprouting. On the 3rd day the highest PAL activity was determined for both untreated and induced with 50 mM H2O2 sprouts obtained by phenylalanine feeding. H2O2 induced TAL activity; the highest TAL activity was determined for 3-day-old sprouts induced with 200 mM H2O2 obtained from seeds fed with phenylalanine.

  16. Two Dynamical Regimes of the Substrate Radical Rearrangement Reaction in B12-Dependent Ethanolamine Ammonia-Lyase Resolve Contributions of Native Protein Configurations and Collective Configurational Fluctuations to Catalysis.

    Science.gov (United States)

    Kohne, Meghan; Zhu, Chen; Warncke, Kurt

    2017-06-27

    The kinetics of the substrate radical rearrangement reaction step in B 12 -dependent ethanolamine ammonia-lyase (EAL) from Salmonella typhimurium are measured over a 92 K temperature range. The observed first-order rate constants display a piecewise-continuous Arrhenius dependence, with linear regions over 295 → 220 K (monoexponential) and 214 → 203 K (biexponential) that are delineated by a kinetic bifurcation and kinks at 219 and 217 K, respectively. The results are interpreted by using a free energy landscape model and derived microscopic kinetic mechanism. The bifurcation and kink transitions correspond to the effective quenching of two distinct sets of native collective protein configurational fluctuations that (1) reconfigure the protein within the substrate radical free energy minimum, in a reaction-enabling step, and (2) create the protein configurations associated with the chemical step. Below 217 K, the substrate radical decay reaction persists. Increases in activation enthalpy and entropy of both the microscopic enabling and reaction steps indicate that this non-native reaction coordinate is conducted by local, incremental fluctuations. Continuity in the Arrhenius relations indicates that the same sets of protein groups and interactions mediate the rearrangement over the 295 to 203 K range, but with a repertoire of configurations below 217 K that is restricted, relative to the native configurations accessible above 219 K. The experimental features of a culled reaction step, first-order kinetic measurements, and wide room-to-cryogenic temperature range, allow the direct demonstration and kinetic characterization of protein dynamical contributions to the core adiabatic, bond-making/bond-breaking reaction in EAL.

  17. The structural model of Salmonella typhimurium ethanolamine ammonia-lyase directs a rational approach to the assembly of the functional [(EutB-EutC)₂]₃ oligomer from isolated subunits.

    Science.gov (United States)

    Bovell, Adonis Miguel; Warncke, Kurt

    2013-02-26

    Ethanolamine ammonia-lyase (EAL) is a 5'-deoxyadenosylcobalamin-dependent bacterial enzyme that catalyzes the deamination of the short-chain vicinal amino alcohols, aminoethanol and (S)- and (R)-2-aminopropanol. The coding sequence for EAL is located within the 17-gene eut operon, which encodes the broad spectrum of proteins that comprise the ethanolamine utilization (eut) metabolosome suborganelle structure. A high-resolution structure of the ∼500 kDa EAL [(EutB-EutC)₂]₃ oligomer from Escherichia coli has been determined by X-ray crystallography, but high-resolution spectroscopic determinations of reactant intermediate-state structures and detailed kinetic and thermodynamic studies of EAL have been conducted for the Salmonella typhimurium enzyme. Therefore, a statistically robust homology model for the S. typhimurium EAL is constructed from the E. coli structure. The model structure is used to describe the hierarchy of EutB and EutC subunit interactions that construct the native EAL oligomer and, specifically, to address the long-standing challenge of reconstitution of the functional oligomer from isolated, purified subunits. Model prediction that the (EutB₂)₃ oligomer assembly will occur from isolated EutB, and that this hexameric structure will template the formation of the complete, native [(EutB-EutC)₂]₃ oligomer, is verified by biochemical methods. Prediction that cysteine residues on the exposed subunit-subunit contact surfaces of isolated EutB and EutC will interfere with assembly by cystine formation is verified by activating effects of disulfide reducing agents. Angstrom-scale congruence of the reconstituted and native EAL in the active site region is shown by electron paramagnetic resonance spectroscopy. Overall, the hierarchy of subunit interactions and microscopic features of the contact surfaces, which are revealed by the homology model, guide and provide a rationale for a refined genetic and biochemical approach to reconstitution of the

  18. Evidence for Improved Encapsulated Pathway Behavior in a Bacterial Microcompartment through Shell Protein Engineering.

    Science.gov (United States)

    Slininger Lee, Marilyn F; Jakobson, Christopher M; Tullman-Ercek, Danielle

    2017-10-20

    Bacterial microcompartments are a class of proteinaceous organelles comprising a characteristic protein shell enclosing a set of enzymes. Compartmentalization can prevent escape of volatile or toxic intermediates, prevent off-pathway reactions, and create private cofactor pools. Encapsulation in synthetic microcompartment organelles will enhance the function of heterologous pathways, but to do so, it is critical to understand how to control diffusion in and out of the microcompartment organelle. To this end, we explored how small differences in the shell protein structure result in changes in the diffusion of metabolites through the shell. We found that the ethanolamine utilization (Eut) protein EutM properly incorporates into the 1,2-propanediol utilization (Pdu) microcompartment, altering native metabolite accumulation and the resulting growth on 1,2-propanediol as the sole carbon source. Further, we identified a single pore-lining residue mutation that confers the same phenotype as substitution of the full EutM protein, indicating that small molecule diffusion through the shell is the cause of growth enhancement. Finally, we show that the hydropathy index and charge of pore amino acids are important indicators to predict how pore mutations will affect growth on 1,2-propanediol, likely by controlling diffusion of one or more metabolites. This study highlights the use of two strategies to engineer microcompartments to control metabolite transport: altering the existing shell protein pore via mutation of the pore-lining residues, and generating chimeras using shell proteins with the desired pores.

  19. Bacterial microcompartments as metabolic modules for plant synthetic biology.

    Science.gov (United States)

    Gonzalez-Esquer, C Raul; Newnham, Sarah E; Kerfeld, Cheryl A

    2016-07-01

    Bacterial microcompartments (BMCs) are megadalton-sized protein assemblies that enclose segments of metabolic pathways within cells. They increase the catalytic efficiency of the encapsulated enzymes while sequestering volatile or toxic intermediates from the bulk cytosol. The first BMCs discovered were the carboxysomes of cyanobacteria. Carboxysomes compartmentalize the enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) with carbonic anhydrase. They enhance the carboxylase activity of RuBisCO by increasing the local concentration of CO2 in the vicinity of the enzyme's active site. As a metabolic module for carbon fixation, carboxysomes could be transferred to eukaryotic organisms (e.g. plants) to increase photosynthetic efficiency. Within the scope of synthetic biology, carboxysomes and other BMCs hold even greater potential when considered a source of building blocks for the development of nanoreactors or three-dimensional scaffolds to increase the efficiency of either native or heterologously expressed enzymes. The carboxysome serves as an ideal model system for testing approaches to engineering BMCs because their expression in cyanobacteria provides a sensitive screen for form (appearance of polyhedral bodies) and function (ability to grow on air). We recount recent progress in the re-engineering of the carboxysome shell and core to offer a conceptual framework for the development of BMC-based architectures for applications in plant synthetic biology. © 2016 The Authors The Plant Journal © 2016 John Wiley & Sons Ltd.

  20. Studies on pectin lyase

    NARCIS (Netherlands)

    Houdenhoven, van F.E.A.

    1975-01-01

    The pectin lyase activity in the commercial enzyme preparation Ultrazym originates from more then one type of enzyme; two of them, accounting for 95 % of the total activity, have been completely purified. As purity criteria specific activity, polyacrylamide disc gel electrophoresis and SDS

  1. Engineering transcriptional regulation to control Pdu microcompartment formation.

    Directory of Open Access Journals (Sweden)

    Edward Y Kim

    Full Text Available Bacterial microcompartments (MCPs show great promise for the organization of engineered metabolic pathways within the bacterial cytoplasm. This subcellular organelle is composed of a protein shell of 100-200 nm diameter that natively encapsulates multi-enzyme pathways. The high energy cost of synthesizing the thousands of protein subunits required for each MCP demands precise regulation of MCP formation for both native and engineered systems. Here, we study the regulation of the propanediol utilization (Pdu MCP, for which growth on 1,2-propanediol induces expression of the Pdu operon for the catabolism of 1,2-propanediol. We construct a fluorescence-based transcriptional reporter to investigate the activation of the Ppdu promoter, which drives the transcription of 21 pdu genes. Guided by this reporter, we find that MCPs can be expressed in strains grown in rich media, provided that glucose is not present. We also characterize the response of the Ppdu promoter to a transcriptional activator of the pdu operon, PocR, and find PocR to be a necessary component of Pdu MCP formation. Furthermore, we find that MCPs form normally upon the heterologous expression of PocR even in the absence of the natural inducer 1,2-propanediol and in the presence of glucose, and that Pdu MCPs formed in response to heterologous PocR expression can metabolize 1,2-propanediol in vivo. We anticipate that this technique of overexpressing a key transcription factor may be used to study and engineer the formation, size, and/or number of MCPs for the Pdu and related MCP systems.

  2. Phenylalanine ammonia lyase from Arabidopsis thaliana (AtPAL2): A potent MIO-enzyme for the synthesis of non-canonical aromatic alpha-amino acids: Part I: Comparative characterization to the enzymes from Petroselinum crispum (PcPAL1) and Rhodosporidium toruloides (RtPAL).

    Science.gov (United States)

    Dreßen, Alana; Hilberath, Thomas; Mackfeld, Ursula; Billmeier, Arne; Rudat, Jens; Pohl, Martina

    2017-09-20

    Phenylalanine ammonia lyase (PAL) from Arabidopsis thaliana (AtPAL2) was comparatively characterized to the well-studied enzyme from parsley (PcPAL1) and Rhodosporidium toruloides (RtPAL) with respect to kinetic parameters for the deamination and the amination reaction, pH- and temperature optima and the substrate range of the amination reaction. Whereas both plant enzymes are specific for phenylalanine, the bifunctional enzyme from Rhodosporidium toruloides shows K M -values for L-Phe and L-Tyr in the same order of magnitude and, compared to both plant enzymes, a 10-15-fold higher activity. At 30°C all enzymes were sufficiently stable with half-lives of 3.4days (PcPAL1), 4.6days (AtPAL2) and 9.7days (RtPAL/TAL). Very good results for the amination of various trans-cinnamic acid derivatives were obtained using E. coli cells as whole cell biocatalysts in ammonium carbonate buffer. Investigation of the substrate ranges gave interesting results for the newly tested enzymes from A. thaliana and R. toruloides. Only the latter accepts besides 4-hydroxy-CA also 3-methoxy-4-hydroxy-CA as a substrate, which is an interesting intermediate for the formation of pharmaceutically relevant L-Dopa. AtPAL2 is a very good catalyst for the formation of (S)-3-F-Phe, (S)-4-F-Phe and (S)-2-Cl-Phe. Such non-canonical amino acids are valuable building blocks for the formation of various drug molecules. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Phenylalanine ammonia lyase from Arabidopsis thaliana (AtPAL2): A potent MIO-enzyme for the synthesis of non-canonical aromatic alpha-amino acids.: Part II: Application in different reactor concepts for the production of (S)-2-chloro-phenylalanine.

    Science.gov (United States)

    Dreßen, Alana; Hilberath, Thomas; Mackfeld, Ursula; Rudat, Jens; Pohl, Martina

    2017-09-20

    Phenylalanine ammonia lyase (PAL) from Arabidopsis thaliana (AtPAL2) is in general a very good catalyst for the amination of fluoro- and chloro-cinnamic acid derivatives yielding halogenated (S)-phenylalanine derivatives with ≥85% conversion and excellent ee values >99%. We have studied the application of this enzyme as whole cell biocatalyst and immobilized on the cellulose carrier Avicel ® for the production of the hypertension drug precursor (S)-2-chloro-phenylalanine using batch, fed-batch, as well as continuous membrane reactor and plug-flow reactor. For immobilization, a C-terminal fusion of the enzyme with a carbohydrate binding module (CBM) was produced, which selectively binds to Avicel ® directly from crude cell extracts, thus enabling a fast and cheap immobilization, stabilization and recycling of the enzyme. 1g Avicel was loaded with 10mg enzyme. Best results were obtained with whole cells using the continuous membrane reactor (47g product /g DryCellWeight ) and using the immobilized enzyme in a repetitive fed-batch (274g product /g immobilized enzyme ) or in a continuous plug-flow reactor (288g product /g immobilize enzyme ). Therewith the productivity of AtPAL2 outperforms the established fed-batch process at DSM using PAL from Rhodotorula glutinis in E. coli as whole cell biocatalyst with a productivity of 0.14g product /g WetCellWeight (ca. 0.7g product /g DryCellWeight ) (de Lange et al., 2011; doi:10.1002/cctc.201000435). Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Adenylosuccinate lyase deficiency.

    Science.gov (United States)

    Jurecka, Agnieszka; Zikanova, Marie; Kmoch, Stanislav; Tylki-Szymańska, Anna

    2015-03-01

    Adenylosuccinate lyase ADSL) deficiency is a defect of purine metabolism affecting purinosome assembly and reducing metabolite fluxes through purine de novo synthesis and purine nucleotide recycling pathways. Biochemically this defect manifests by the presence in the biologic fluids of two dephosphorylated substrates of ADSL enzyme: succinylaminoimidazole carboxamide riboside (SAICAr) and succinyladenosine (S-Ado). More than 80 individuals with ADSL deficiency have been identified, but incidence of the disease remains unknown. The disorder shows a wide spectrum of symptoms from slowly to rapidly progressing forms. The fatal neonatal form has onset from birth and presents with fatal neonatal encephalopathy with a lack of spontaneous movement, respiratory failure, and intractable seizures resulting in early death within the first weeks of life. Patients with type I (severe form) present with a purely neurologic clinical picture characterized by severe psychomotor retardation, microcephaly, early onset of seizures, and autistic features. A more slowly progressing form has also been described (type II, moderate or mild form), as having later onset, usually within the first years of life, slight to moderate psychomotor retardation and transient contact disturbances. Diagnosis is facilitated by demonstration of SAICAr and S-Ado in extracellular fluids such as plasma, cerebrospinal fluid and/or followed by genomic and/or cDNA sequencing and characterization of mutant proteins. Over 50 ADSL mutations have been identified and their effects on protein biogenesis, structural stability and activity as well as on purinosome assembly were characterized. To date there is no specific and effective therapy for ADSL deficiency.

  5. Argininosuccinate lyase deficiency.

    Science.gov (United States)

    Nagamani, Sandesh C S; Erez, Ayelet; Lee, Brendan

    2012-05-01

    The urea cycle consists of six consecutive enzymatic reactions that convert waste nitrogen into urea. Deficiencies of any of these enzymes of the cycle result in urea cycle disorders (UCDs), a group of inborn errors of hepatic metabolism that often result in life-threatening hyperammonemia. Argininosuccinate lyase (ASL) catalyzes the fourth reaction in this cycle, resulting in the breakdown of argininosuccinic acid to arginine and fumarate. ASL deficiency (ASLD) is the second most common UCD, with a prevalence of ~1 in 70,000 live births. ASLD can manifest as either a severe neonatal-onset form with hyperammonemia within the first few days after birth or as a late-onset form with episodic hyperammonemia and/or long-term complications that include liver dysfunction, neurocognitive deficits, and hypertension. These long-term complications can occur in the absence of hyperammonemic episodes, implying that ASL has functions outside of its role in ureagenesis and the tissue-specific lack of ASL may be responsible for these manifestations. The biochemical diagnosis of ASLD is typically established with elevation of plasma citrulline together with elevated argininosuccinic acid in the plasma or urine. Molecular genetic testing of ASL and assay of ASL enzyme activity are helpful when the biochemical findings are equivocal. However, there is no correlation between the genotype or enzyme activity and clinical outcome. Treatment of acute metabolic decompensations with hyperammonemia involves discontinuing oral protein intake, supplementing oral intake with intravenous lipids and/or glucose, and use of intravenous arginine and nitrogen-scavenging therapy. Dietary restriction of protein and dietary supplementation with arginine are the mainstays in long-term management. Orthotopic liver transplantation (OLT) is best considered only in patients with recurrent hyperammonemia or metabolic decompensations resistant to conventional medical therapy.

  6. Comparative activities of phenylalanine ammonia-lyase and tyrosine ...

    African Journals Online (AJOL)

    Plants respond to attack by pathogens by initiating a change in cellular metabolism, leading to synthesis of antifungal proteins, production of phytoalexins and/or accumulation of phenolic compounds, namely lignins and salicylic. Lignins reinforce pectocellulosic cell walls and limit the invasion of plant tissues by pathogens; ...

  7. Molecular analysis and expression of phenylalanine ammonia-lyase ...

    African Journals Online (AJOL)

    Meanwhile, expression levels increased in the order: green leaves - turning color leaves - bracts, which were consistent with their anthocyanin content during growth and development of bracts. The curve of diurnal variation of EpPAL expression level in bracts exhibited two highest peaks at 9:00 and 18:00, respectively, and ...

  8. Involvement of a bacterial microcompartment in the metabolism of fucose and rhamnose by Clostridium phytofermentans.

    Directory of Open Access Journals (Sweden)

    Elsa Petit

    Full Text Available Clostridium phytofermentans, an anaerobic soil bacterium, can directly convert plant biomass into biofuels. The genome of C. phytofermentans contains three loci with genes encoding shell proteins of bacterial microcompartments (BMC, organelles composed entirely of proteins.One of the BMC loci has homology to a BMC-encoding locus implicated in the conversion of fucose to propanol and propionate in a human gut commensal, Roseburia inulinivorans. We hypothesized that it had a similar role in C. phytofermentans. When C. phytofermentans was grown on fucose, the major products identified were ethanol, propanol and propionate. Transmission electron microscopy of fucose- and rhamnose-grown cultures revealed polyhedral structures, presumably BMCs. Microarray analysis indicated that during growth on fucose, operons coding for the BMC locus, fucose dissimilatory enzymes, and an ATP-binding cassette transporter became the dominant transcripts. These data are consistent with fucose fermentation producing a 1,2-propanediol intermediate that is further metabolized in the microcompartment encoded in the BMC locus. Growth on another deoxyhexose sugar, rhamnose, resulted in the expression of the same BMC locus and similar fermentation products. However, a different set of dissimilatory enzymes and transport system genes were induced. Quite surprisingly, growth on fucose or rhamnose also led to the expression of a diverse array of complex plant polysaccharide-degrading enzymes.Based on physiological, genomic, and microarray analyses, we propose a model for the fermentation of fucose and rhamnose in C. phytofermentans that includes enzymes encoded in the same BMC locus. Comparative genomic analysis suggests that this BMC may be present in other clostridial species.

  9. Bacterial Microcompartments

    Energy Technology Data Exchange (ETDEWEB)

    Kerfeld, Cheryl A.; Heinhorst, Sabine; Cannon, Gordon C.

    2010-06-05

    Bacterialmicrocompartments (BMCs) are organelles composed entirely of protein. They promote specific metabolic processes by encapsulatingand colocalizing enzymes with their substrates and cofactors, by protecting vulnerable enzymes in a defined microenvironment, and bysequestering toxic or volatile intermediates. Prototypes of the BMCsare the carboxysomes of autotrophic bacteria. However, structures of similarpolyhedral shape are being discovered in an ever-increasing number of heterotrophic bacteria, where they participate in the utilization ofspecialty carbon and energy sources.Comparative genomics reveals that the potential for this type of compartmentalization is widespread acrossbacterial phyla and suggests that genetic modules encoding BMCs are frequently laterally transferred among bacteria. The diverse functionsof these BMCs suggest that they contribute to metabolic innovation in bacteria in a broad range of environments.

  10. Ammonia intoxication

    International Nuclear Information System (INIS)

    Bessman, S.P.; Pal, N.

    1982-01-01

    Data is presented which shows that there is a relation between ammonia concentration in the blood and state of consciousness. The concentrations of GTP and ATP also relate both to the ammonia concentration in blood and the state of consciousness. The rate of protein synthesis in the brain as measured by the percent of intracellular counts that are incorporated into protein is also related to ammonia concentration. These findings of energy depletion and depressed synthesis resulting from energy depletion suggest that the primary lesion in ammonia intoxication involves the Krebs cycle. The greater effect of ammonia on GTP than on ATP metabolism supports the view that the primary site of action of ammonia is at the glutamate dehydrogenase-ketoglutarate reduction step - and is consistent with previous work on this subject. (H.K.)

  11. Ammonia Monitor

    Science.gov (United States)

    Sauer, Richard L. (Inventor); Akse, James R. (Inventor); Thompson, John O. (Inventor); Atwater, James E. (Inventor)

    1999-01-01

    Ammonia monitor and method of use are disclosed. A continuous, real-time determination of the concentration of ammonia in an aqueous process stream is possible over a wide dynamic range of concentrations. No reagents are required because pH is controlled by an in-line solid-phase base. Ammonia is selectively transported across a membrane from the process stream to an analytical stream to an analytical stream under pH control. The specific electrical conductance of the analytical stream is measured and used to determine the concentration of ammonia.

  12. Ammonia blood test

    Science.gov (United States)

    ... page: //medlineplus.gov/ency/article/003506.htm Ammonia blood test To use the sharing features on this page, ... Encephalopathy - ammonia; Cirrhosis - ammonia; Liver failure - ammonia Images Blood test References Chernecky CC, Berger BJ. Ammonia (NH3) - blood ...

  13. Bacterial microcompartments: widespread prokaryotic organelles for isolation and optimization of metabolic pathways.

    Science.gov (United States)

    Bobik, Thomas A; Lehman, Brent P; Yeates, Todd O

    2015-10-01

    Prokaryotes use subcellular compartments for a variety of purposes. An intriguing example is a family of complex subcellular organelles known as bacterial microcompartments (MCPs). MCPs are widely distributed among bacteria and impact processes ranging from global carbon fixation to enteric pathogenesis. Overall, MCPs consist of metabolic enzymes encased within a protein shell, and their function is to optimize biochemical pathways by confining toxic or volatile metabolic intermediates. MCPs are fundamentally different from other organelles in having a complex protein shell rather than a lipid-based membrane as an outer barrier. This unusual feature raises basic questions about organelle assembly, protein targeting and metabolite transport. In this review, we discuss the three best-studied MCPs highlighting atomic-level models for shell assembly, targeting sequences that direct enzyme encapsulation, multivalent proteins that organize the lumen enzymes, the principles of metabolite movement across the shell, internal cofactor recycling, a potential system of allosteric regulation of metabolite transport and the mechanism and rationale behind the functional diversification of the proteins that form the shell. We also touch on some potential biotechnology applications of an unusual compartment designed by nature to optimize metabolic processes within a cellular context. © 2015 John Wiley & Sons Ltd.

  14. Rhamnogalacturonan lyase reveals a unique three-domain modular structure for polysaccharide lyase family 4

    DEFF Research Database (Denmark)

    McDonough, Michael A.; Kadirvelraj, Renuka; Harris, Pernille

    2004-01-01

    Rhamnogalacturonan lyase (RG-lyase) specifically recognizes and cleaves alpha-1,4 glycosidic bonds between L-rhamnose and D-galacturonic acids in the backbone of rhamno galacturonan-I, a major component of the plant cell wall polysaccharide, pectin. The three-dimensional structure of RG-lyase from...... Aspergillus aculeatus has been determined to 1.5 Angstrom resolution representing the first known structure from polysaccharide lyase family 4 and of an enzyme with this catalytic specificity. The 508-amino acid polypeptide displays a unique arrangement of three distinct modular domains. Each domain shows...

  15. Prenatal diagnosis in adenylosuccinate lyase deficiency

    NARCIS (Netherlands)

    Marie, S.; Flipsen, J. W.; Duran, M.; Poll-The, B. T.; Beemer, F. A.; Bosschaart, A. N.; Vincent, M. F.; van den Berghe, G.

    2000-01-01

    Adenylosuccinate lyase deficiency, an autosomal recessive inborn error of purine synthesis, provokes accumulation in body fluids of succinylaminoimidazolecarboxamide riboside and succinyladenosine, the dephosphorylated derivatives of the two substrates of the enzyme. Most patients display severe

  16. Enhancing RGI lyase thermostability by targeted single point mutations

    DEFF Research Database (Denmark)

    Silva, Inês R.; Larsen, Dorte Møller; Jers, Carsten

    2013-01-01

    Rhamnogalacturonan I lyase (RGI lyase) (EC 4.2.2.-) catalyzes the cleavage of rhamnogalacturonan I in pectins by β-elimination. In this study the thermal stability of a RGI lyase (PL 11) originating from Bacillus licheniformis DSM 13/ATCC14580 was increased by a targeted protein engineering...

  17. Misleading behavioural phenotype with adenylosuccinate lyase deficiency.

    Science.gov (United States)

    Gitiaux, Cyril; Ceballos-Picot, Irène; Marie, Sandrine; Valayannopoulos, Vassili; Rio, Marlène; Verrieres, Séverine; Benoist, Jean François; Vincent, Marie Françoise; Desguerre, Isabelle; Bahi-Buisson, Nadia

    2009-01-01

    Adenylosuccinate lyase deficiency is a rare autosomal disorder of de novo purine synthesis, which results in the accumulation of succinylpurines in body fluids. Patients with adenylosuccinate lyase deficiency show a variable combination of mental retardation, epilepsy and autistic features and are usually discovered during screens for unexplained encephalopathy using the Bratton-Marshall assay that reveals the excretion of the succinylaminoimidazolecarboxamide riboside (SAICAr). Here, we report on two sisters aged 11 and 12 years presented with global developmental delay, motor apraxia, severe speech deficits, seizures and behavioural features, which combined excessive laughter, a very happy disposition, hyperactivity, a short attention span, the mouthing of objects, tantrums and stereotyped movements that gave a behavioural profile mimicking Angelman syndrome. Both patients had an increased succinyladenosine/SAICAr ratio of 1.6, and exhibited a novel homozygous missense mutation (c.674T>C; p.Met225Thr) in the exon 6 of the ADSL gene. We suggest that these clinical features might be a new presentation of adenylosuccinate lyase deficiency. On the basis of this observation, although adenylosuccinate lyase deficiency is a rare disorder, this diagnosis should be considered in patients with mental retardation and a behavioural profile suggestive of Angelman syndrome.

  18. Exploring Lactobacillus reuteri DSM20016 as a biocatalyst for transformation of longer chain 1,2-diols: Limits with microcompartment.

    Science.gov (United States)

    Chen, Lu; Hatti-Kaul, Rajni

    2017-01-01

    Lactobacillus reuteri metabolises glycerol efficiently to form 3-hydroxypropionic acid (3-HP) and 1,3-propanediol (1,3PDO) by the same mechanism as that for 1,2-propanediol (1,2PDO) conversion to propionic acid and propanol via its propanediol utilization (pdu) pathway. Pdu enzymes are encoded by the pdu-operon, which also contain genes encoding the shell proteins of the microcompartment housing the metabolic pathway. In this work the selectivity and kinetics of the reactions catalysed by L. reuteri DSM20016 Pdu enzymes glycerol dehydratase (GDH), 1,3-propanediol oxidoreductase (PduQ) and coenzyme-A acylating propionaldehyde dehydrogenase (PduP), produced recombinantly, was investigated against corresponding substrates of different chain lengths. Glycerol dehydratase exhibited activity against C2-C4 polyols, with the highest activity against glycerol and 1,2-propanediol (1,2-PDO). A double mutant of the pduC gene of GDH (PduC-S302A/Q337A) was constructed that displayed lowered activity against glycerol and 1,2PDO but extended the substrate range upto C6-diol. The best substrate for both PduQ and PduP was 3-hydroxypropanal (3HPA), although PduP exhibited nearly 10-fold higher specific activity. The enzymes also showed some activity against C3-C10 aliphatic aldehydes, with PduP having higher relative activity. Subsequently, transformation of polyols using whole cells of L. reuteri containing the wild type- and mutated GDH, respectively, confirmed the reduced activity of the mutant against glycerol and 1,2PDO, but its activity against longer substrates was negligible. In contrast, recombinant Escherichia coli BL21(DE3) cells harboring the GDH variant converted diols with up to C6 carbon chain length to their respective aldehydes, suggesting that the protein shell of the microcompartment in L. reuteri posed a barrier to the passage of longer chain substrate.

  19. Characterisation of PduS, the pdu metabolosome corrin reductase, and evidence of substructural organisation within the bacterial microcompartment.

    Directory of Open Access Journals (Sweden)

    Joshua B Parsons

    2010-11-01

    Full Text Available PduS is a corrin reductase and is required for the reactivation of the cobalamin-dependent diol dehydratase. It is one component encoded within the large propanediol utilisation (pdu operon, which is responsible for the catabolism of 1,2-propanediol within a self-assembled proteinaceous bacterial microcompartment. The enzyme is responsible for the reactivation of the cobalamin coenzyme required by the diol dehydratase. The gene for the cobalamin reductase from Citrobacter freundii (pduS has been cloned to allow the protein to be overproduced recombinantly in E. coli with an N-terminal His-tag. Purified recombinant PduS is shown to be a flavoprotein with a non-covalently bound FMN that also contains two coupled [4Fe-4S] centres. It is an NADH-dependent flavin reductase that is able to mediate the one-electron reductions of cob(IIIalamin to cob(IIalamin and cob(IIalamin to cob(Ialamin. The [4Fe-4S] centres are labile to oxygen and their presence affects the midpoint redox potential of flavin. Evidence is presented that PduS is able to bind cobalamin, which is inconsistent with the view that PduS is merely a flavin reductase. PduS is also shown to interact with one of the shell proteins of the metabolosome, PduT, which is also thought to contain an [Fe-S] cluster. PduS is shown to act as a corrin reductase and its interaction with a shell protein could allow for electron passage out of the bacterial microcompartment.

  20. Ligand-induced type II interleukin-4 receptor dimers are sustained by rapid re-association within plasma membrane microcompartments

    Science.gov (United States)

    Richter, David; Moraga, Ignacio; Winkelmann, Hauke; Birkholz, Oliver; Wilmes, Stephan; Schulte, Markos; Kraich, Michael; Kenneweg, Hella; Beutel, Oliver; Selenschik, Philipp; Paterok, Dirk; Gavutis, Martynas; Schmidt, Thomas; Garcia, K. Christopher; Müller, Thomas D.; Piehler, Jacob

    2017-07-01

    The spatiotemporal organization of cytokine receptors in the plasma membrane is still debated with models ranging from ligand-independent receptor pre-dimerization to ligand-induced receptor dimerization occurring only after receptor uptake into endosomes. Here, we explore the molecular and cellular determinants governing the assembly of the type II interleukin-4 receptor, taking advantage of various agonists binding the receptor subunits with different affinities and rate constants. Quantitative kinetic studies using artificial membranes confirm that receptor dimerization is governed by the two-dimensional ligand-receptor interactions and identify a critical role of the transmembrane domain in receptor dimerization. Single molecule localization microscopy at physiological cell surface expression levels, however, reveals efficient ligand-induced receptor dimerization by all ligands, largely independent of receptor binding affinities, in line with the similar STAT6 activation potencies observed for all IL-4 variants. Detailed spatiotemporal analyses suggest that kinetic trapping of receptor dimers in actin-dependent microcompartments sustains robust receptor dimerization and signalling.

  1. Alanine Scanning Mutagenesis Identifies an Asparagine–Arginine–Lysine Triad Essential to Assembly of the Shell of the Pdu Microcompartment

    Energy Technology Data Exchange (ETDEWEB)

    Sinha, Sharmistha; Cheng, Shouqiang; Sung, Yea Won; McNamara, Dan E.; Sawaya, Michael R.; Yeates, Todd O.; Bobik, Thomas A.

    2014-06-01

    Bacterial microcompartments (MCPs) are the simplest organelles known. They function to enhance metabolic pathways by confining several related enzymes inside an all-protein envelope called the shell. In this study, we investigated the factors that govern MCP assembly by performing scanning mutagenesis on the surface residues of PduA, a major shell protein of the MCP used for 1,2-propanediol degradation. Biochemical, genetic, and structural analysis of 20 mutants allowed us to determine that PduA K26, N29, and R79 are crucial residues that stabilize the shell of the 1,2-propanediol MCP. In addition, we identify two PduA mutants (K37A and K55A) that impair MCP function most likely by altering the permeability of its protein shell. These are the first studies to examine the phenotypic effects of shell protein structural mutations in an MCP system. The findings reported here may be applicable to engineering protein containers with improved stability for biotechnology applications.

  2. Xylella fastidiosa esterase rather than hydroxynitrile lyase.

    Science.gov (United States)

    Torrelo, Guzman; Ribeiro de Souza, Fayene Zeferino; Carrilho, Emanuel; Hanefeld, Ulf

    2015-03-02

    In 2009, we reported that the product of the gene SCJ21.16 (XFa0032) from Xylella fastidiosa, a xylem-restricted plant pathogen that causes a range of diseases in several important crops, encodes a protein (XfHNL) with putative hydroxynitrile lyase activity. Sequence analysis and activity tests indicated that XfHNL exhibits an α/β-hydrolase fold and could be classified as a member of the family of FAD-independent HNLs. Here we provide a more detailed sequence analysis and new experimental data. Using pure heterologously expressed XfHNL we show that this enzyme cannot catalyse the cleavage/synthesis of mandelonitrile and that this protein is in fact a non-enantioselective esterase. Homology modelling and ligand docking simulations were used to study the active site and support these results. This finding could help elucidate the common ancestor of esterases and hydroxynitrile lyases with an α/β -hydrolase fold. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Mammalian O-phosphorylethanolamine phospho-lyase activity and its inhibition.

    Science.gov (United States)

    Grøn, I H

    1978-04-01

    The activity of the enzyme O-phosphorylethanolamine phospho-lyase, metabolizing O-phosphorylethanolamine to acetaldehyde, orthophosphate, and ammonia in vitro, was studied in human liver biopsy and autopsy material, and leucocytes. Only in the liver biopsies enzyme activity towards O-phosphorylethanolamine could be found, and in amounts corresponding to one tenth of the activity found in rat liver examined under identical conditions. The enzyme activity of the liver biopsies was confined to the post-microsomal fraction, the activity amounting to 35 +/- 7 (SD) micromicron/mg protein. The results suggest the presence of an inhibiting factor of protein character. Inhibition was not due to competition from alkaline phosphatase (E.C. 3.1.3.1.) or O-phosphorylethanolamine cytidylyl-transferase (E.C. 2.7.7.14).

  4. Aquatic Life Criteria - Ammonia

    Science.gov (United States)

    Documents related to EPA's final 2013 Aquatic Life Ambient Water Quality Criteria for Ammonia (Freshwater). These documents pertain to the safe levels of Ammonia in water that should protect to the majority of species.

  5. Adenylosuccinate lyase deficiency--first British case.

    Science.gov (United States)

    Marinaki, A M; Champion, M; Kurian, M A; Simmonds, H A; Marie, S; Vincent, M F; van den Berghe, G; Duley, J A; Fairbanks, L D

    2004-10-01

    A deficiency of adenylosuccinate lyase (ASDL) is characterised by the accumulation of SAICAriboside (SAICAr) and succinyladenosine (S-Ado) in body fluids. The severity of the clinical presentation correlates with a low S-Ado/SAICAr ratio in body fluids. We report the first British case of ADSL deficiency. The patient presented at 14 days with a progressive neonatal encephalopathy and seizures. There was marked axial and peripheral hypotonia. Brain MRI showed widespread white matter changes. She died at 4 weeks of age. Concentrations of SAICAr and SAdo were markedly elevated in urine, plasma and CSF and the SAdo/SAICAr ratio was low, consistent with the severe phenotype. The patient was compound heterozygous for 2 novel ADSL mutations; c.9 G>C (A3P) and c.572 C>T (R190X).

  6. Novel proton MR spectroscopy findings in adenylosuccinate lyase deficiency

    NARCIS (Netherlands)

    Zulfiqar, M.; Lin, D.D.; Graaf, M. van der; Barker, P.B.; Fahrner, J.A.; Marie, S. de; Morava, E.; Boer, L. de; Willemsen, M.A.A.P.; Vining, E.; Horska, A.; Engelke, U.F.H.; Wevers, R.A.; Maegawa, G.H.

    2013-01-01

    Adenylosuccinate lyase (ADSL) deficiency is a rare inborn error of metabolism resulting in accumulation of metabolites including succinylaminoimidazole carboxamide riboside (SAICAr) and succinyladenosine (S-Ado) in the brain and other tissues. Patients with ADSL have progressive psychomotor

  7. The hydroxynitrile lyase from almond: crystal structure and mechanistical studies

    International Nuclear Information System (INIS)

    Dreveny, Ingrid

    2001-09-01

    Cyanogenesis is a defense process of several thousand plant species. Hydroxynitrile lyase (HNL), a key enzyme of this process, cleaves a cyanohydrin precursor into hydrocyanic acid and the corresponding aldehyde or ketone. The reverse reaction constitutes an important tool in industrial biocatalysis. Different classes of hydroxynitrile lyases have convergently evolved from FAD-dependent oxidoreductases, α/β hydrolases and alcohol dehydrogenases. The FAD-dependent hydroxynitrile lyases (FAD-HNLs) carry a flavin cofactor whose redox properties appear to be unimportant for catalysis. The high resolution crystal structure of the hydroxynitrile lyase from almond (Prunus amygdalus), PaHNL1, has been determined and constitutes the first 3D structure of an FAD-HNL. The overall fold and the architecture of the active site region showed that PaHNL1 belongs to the glucose-methanol-choline-oxidoreductase family, with closest structural similarity to glucose oxidase. There is strong evidence from the sequence and the reaction product that FAD-dependent hydroxynitrile lyases have evolved from an aryl alcohol oxidizing precursor. Structures of PaHNL1 in complex with its natural substrate mandelonitrile and the competitive inhibitor benzyl alcohol provided insight into the residues involved in catalysis and a mechanism without participation of the cofactor could be suggested. Although the catalytic residues differ between the α/β-hydrolase-type HNLs and PaHNL1, common general features relevant for hydroxynitrile lyase activity could be proposed. (author)

  8. Sources of atmospheric ammonia

    International Nuclear Information System (INIS)

    Harriss, R.C.; Michaels, J.T.

    1982-01-01

    The information available on factors that influence emissions from the principal societal sources of ammonia to the atmosphere, namely combustion processes, volatilization of farm animal wastes, and volatilization of fertilizers, is reviewed. Emission factors are established for each major source of atmospheric ammonia. The factors are then multiplied by appropriate source characterization descriptors to obtain calculated fluxes of ammonia to the atmosphere on a state-by-state basis for the United States

  9. Catalyst for Ammonia Oxidation

    DEFF Research Database (Denmark)

    2015-01-01

    The present invention relates to a bimetallic catalyst for ammonia oxidation, a method for producing a bimetallic catalyst for ammonia oxidation and a method for tuning the catalytic activity of a transition metal. By depositing an overlayer of less catalytic active metal onto a more catalytic...

  10. Method for forming ammonia

    Science.gov (United States)

    Kong, Peter C.; Pink, Robert J.; Zuck, Larry D.

    2008-08-19

    A method for forming ammonia is disclosed and which includes the steps of forming a plasma; providing a source of metal particles, and supplying the metal particles to the plasma to form metal nitride particles; and providing a substance, and reacting the metal nitride particles with the substance to produce ammonia, and an oxide byproduct.

  11. Expression of Phenylalanine Ammonia Lyase Genes in Maize Lines Differing in Susceptibility to Meloidogyne incognita

    OpenAIRE

    Starr, J. L.; Yang, W.; Yan, Y.; Crutcher, F.; Kolomiets, M.

    2014-01-01

    Maize is a well-known host for Meloidogyne incognita, and there is substantial variation in host status among maize genotypes. In previous work it was observed that nematode reproduction increased in the moderately susceptible maize inbred line B73 when the ZmLOX3 gene from oxylipid metabolism was knocked out. Additionally, in this mutant line, use of a nonspecific primer for phenyl alanine ammonialyase (PAL) genes indicated that expression of these genes was reduced in the mutant maize plant...

  12. Expression of Phenylalanine Ammonia Lyase Genes in Maize Lines Differing in Susceptibility to Meloidogyne incognita.

    Science.gov (United States)

    Starr, J L; Yang, W; Yan, Y; Crutcher, F; Kolomiets, M

    2014-12-01

    Maize is a well-known host for Meloidogyne incognita, and there is substantial variation in host status among maize genotypes. In previous work it was observed that nematode reproduction increased in the moderately susceptible maize inbred line B73 when the ZmLOX3 gene from oxylipid metabolism was knocked out. Additionally, in this mutant line, use of a nonspecific primer for phenyl alanine ammonialyase (PAL) genes indicated that expression of these genes was reduced in the mutant maize plants whereas expression of several other defense related genes was increased. In this study, we used more specific gene primers to examine the expression of six PAL genes in three maize genotypes that were good, moderate, and poor hosts for M. incognita, respectively. Of the six PAL genes interrogated, two (ZmPAL3 and ZmPAL6) were not expressed in either M. incognita-infected or noninfected roots. Three genes (ZmPAL1, ZmPAL2, and ZmPAL5) were strongly expressed in all three maize lines, in both nematode-infected and noninfected roots, between 2 and 16 d after inoculation (DAI). In contrast, ZmPAL4 was most strongly expressed in the most-resistant maize line W438, was not detected in the most-susceptible maize line CML, and was detected only at 8 DAI in the maize line B73 that supported intermediate levels of reproduction by M. incognita. These observations are consistent with at least one PAL gene playing a role in modulating host status of maize toward M. incognita and suggest a need for additional research to further elucidate this association.

  13. Phenylalanine Ammonia Lyase Activity in Stem of Pepper (Capsicum annuum L.) Infected by Phytophthora capsici L.

    OpenAIRE

    KOC, Esra; USTUN, Ayşen Sulun

    2012-01-01

    In this study, PAL activity in stems of pepper (Capsicum annuum L.) plants infected by the root rot pathogen Phytophthora capsici-22 in a resistant (PM-702) and two susceptible (Kahramanmaraş-Hot (KM-Hot) and Demre-8) cultivar were studied. The response of the PAL activity in the resistant cultivar was faster and higher than in the susceptible cultivars (p < 0.01). The increase in production of  PAL upon Phytophthora capsici-22 were higher in the infected plants compared to the non-in...

  14. Structural and Functional Characterization of a Short-Chain Flavodoxin Associated with a Noncanonical 1,2-Propanediol Utilization Bacterial Microcompartment

    Energy Technology Data Exchange (ETDEWEB)

    Plegaria, Jefferson S. [MSU-DOE; Sutter, Markus [MSU-DOE; Molecular; Ferlez, Bryan [MSU-DOE; Aussignargues, Clément [MSU-DOE; Niklas, Jens [Solar; Poluektov, Oleg G. [Solar; Fromwiller, Ciara [MSU-DOE; TerAvest, Michaela [Department; amp, Molecular Biology, Michigan State University, East; Utschig, Lisa M. [Solar; Tiede, David M. [Solar; Kerfeld, Cheryl A. [MSU-DOE; Molecular; Department; amp, Molecular Biology, Michigan State University, East; Berkeley Synthetic Biology Institute, Berkeley, California 94720, United States

    2017-09-21

    Bacterial microcompartments (BMCs) are proteinaceous organelles that encapsulate enzymes involved in CO2 fixation (carboxysomes). or carbon catabolism (metabolosomes). Metabolosomes share a common core of enzymes and a distinct signature enzyme for substrate degradation that defines the function of the BMC (e,g., propanediol or ethanolamine utilization BMCs, or glycyl-radical enzyme microcompartments). Loci encoding metabolosomes also typically contain genes for proteins that support organelle function, such as regulation, transport of substrate, and cofactor (e.g., vitamin B-12) synthesis and recycling. Flavoproteins are frequently among these ancillary gene products, suggesting that these redox active proteins play an undetermined function in many metabolosomes. Here, we report the first characterization of a BMC-associated flavodoxin (Fld1C), a small flavoprotein, derived from the noncanonical 1,2-propanediol utilization BMC locus (PDU1C) of Lactobacillus reuteri. The 2.0 angstrom X-ray structure of Fld1C displays the alpha/beta flavodoxin fold, which noncovalently binds a single flavin mononucleotide molecule. Fld1C is a short-chain flavodoxin with redox potentials of -240 +/- 3 mV oxidized/semiquinone and -344 +/- 1 mV semiquinone/hydroquinone versus the standard hydrogen electrode at pH 7.5. It can participate in an electron transfer reaction with a photoreductant to form a stable semiquinone species. Collectively, our structural and functional results suggest that PDU1C BMCs encapsulate Fld1C to store and transfer electrons for the reactivation and/or recycling of the B-12 cofactor utilized by the signature enzyme.

  15. Ammonia Release on ISS

    Science.gov (United States)

    Macatangay, Ariel

    2009-01-01

    Crew: Approximately 53% metabolic load Product of protein metabolism Limit production of ammonia by external regulation NOT possbile Payloads Potential source Scientific experiments Thorough safety review ensures sufficient levels of containment

  16. Reactor for removing ammonia

    Science.gov (United States)

    Luo, Weifang [Livermore, CA; Stewart, Kenneth D [Valley Springs, CA

    2009-11-17

    Disclosed is a device for removing trace amounts of ammonia from a stream of gas, particularly hydrogen gas, prepared by a reformation apparatus. The apparatus is used to prevent PEM "poisoning" in a fuel cell receiving the incoming hydrogen stream.

  17. Titan's Ammonia Feature

    Science.gov (United States)

    Smythe, W.; Nelson, R.; Boryta, M.; Choukroun, M.

    2011-01-01

    NH3 has long been considered an important component in the formation and evolution of the outer planet satellites. NH3 is particularly important for Titan, since it may serve as the reservoir for atmospheric nitrogen. A brightening seen on Titan starting in 2004 may arise from a transient low-lying fog or surface coating of ammonia. The spectral shape suggests the ammonia is anhydrous, a molecule that hydrates quickly in the presence of water.

  18. Isolation, expression and comparison of a pectate lyase produced ...

    African Journals Online (AJOL)

    Administrator

    2010-12-27

    Dec 27, 2010 ... Key words: Pectate lyase, Fusarium oxysporum f.sp. cubense, Fusarium wilt, banana. INTRODUCTION. Banana (Musa spp.) is a number four important crop in developing countries (Heslop-Harrison and Schwarzacher,. 2007). Banana production suffers from several diseases and Fusarium wilt disease ...

  19. Cystathionine .gamma.-lyase: Clinical, metabolic, genetic, and structural studies

    Czech Academy of Sciences Publication Activity Database

    Kraus, J. P.; Hašek, Jindřich; Kožich, V.; Collard, R.; Venezia, S.; Janošíková, B.; Wang, J.; Stabler, S. P.; Allen, R. H.; Jakobs, C.; Finn, C. T.; Chien, Y. H.; Hwu, W. L.; Hegele, R. A.; Mudd, S. H.

    2009-01-01

    Roč. 97, č. 4 (2009), s. 250-259 ISSN 1096-7192 R&D Projects: GA ČR GA305/07/1073 Institutional research plan: CEZ:AV0Z40500505 Keywords : cystathionine gamma-lyase * cystathioninuria * hypercystathioninemia Subject RIV: CD - Macromolecular Chemistry Impact factor: 2.897, year: 2009

  20. Catalytic Promiscuity of Ancestral Esterases and Hydroxynitrile Lyases.

    Science.gov (United States)

    Devamani, Titu; Rauwerdink, Alissa M; Lunzer, Mark; Jones, Bryan J; Mooney, Joanna L; Tan, Maxilmilien Alaric O; Zhang, Zhi-Jun; Xu, Jian-He; Dean, Antony M; Kazlauskas, Romas J

    2016-01-27

    Catalytic promiscuity is a useful, but accidental, enzyme property, so finding catalytically promiscuous enzymes in nature is inefficient. Some ancestral enzymes were branch points in the evolution of new enzymes and are hypothesized to have been promiscuous. To test the hypothesis that ancestral enzymes were more promiscuous than their modern descendants, we reconstructed ancestral enzymes at four branch points in the divergence hydroxynitrile lyases (HNL's) from esterases ∼ 100 million years ago. Both enzyme types are α/β-hydrolase-fold enzymes and have the same catalytic triad, but differ in reaction type and mechanism. Esterases catalyze hydrolysis via an acyl enzyme intermediate, while lyases catalyze an elimination without an intermediate. Screening ancestral enzymes and their modern descendants with six esterase substrates and six lyase substrates found higher catalytic promiscuity among the ancestral enzymes (P promiscuous and catalyzed both hydrolysis and lyase reactions with many substrates. A broader screen tested mechanistically related reactions that were not selected for by evolution: decarboxylation, Michael addition, γ-lactam hydrolysis and 1,5-diketone hydrolysis. The ancestral enzymes were more promiscuous than their modern descendants (P = 0.04). Thus, these reconstructed ancestral enzymes are catalytically promiscuous, but HNL1 is especially so.

  1. [Adenylosuccinate lyase deficiency: an unusual cause of neonatal seizure].

    Science.gov (United States)

    Clamadieu, C; Cottin, X; Rousselle, C; Claris, O

    2008-02-01

    Adenylosuccinate lyase deficiency is an autosomal recessive inborn error of purine synthesis, which provokes epilepsy, psychomotor delay and/or autistic features. We report on two siblings with ADSL deficiency, who developed seizures on the first day of life. ADSL deficiency should be part of the screening to be performed in case of neonatal seizures.

  2. Correlation between the cystathionine-r-lyase (CES) and the ...

    African Journals Online (AJOL)

    Background: The infection of Helicobacter pylori (H. pylori) is one of the most important causes of gastric ulcer disease. The role of hydrogen sulfide (H2S) production in H. pylori-induced gastric ulcer disease. Aim: The expression of cystathionine-γ-lyase (CSE) was determined, and correlated with the severity of gastric ulcer ...

  3. In Silico Characterization of Pectate Lyase Protein Sequences from Different Source Organisms

    Directory of Open Access Journals (Sweden)

    Amit Kumar Dubey

    2010-01-01

    Full Text Available A total of 121 protein sequences of pectate lyases were subjected to homology search, multiple sequence alignment, phylogenetic tree construction, and motif analysis. The phylogenetic tree constructed revealed different clusters based on different source organisms representing bacterial, fungal, plant, and nematode pectate lyases. The multiple accessions of bacterial, fungal, nematode, and plant pectate lyase protein sequences were placed closely revealing a sequence level similarity. The multiple sequence alignment of these pectate lyase protein sequences from different source organisms showed conserved regions at different stretches with maximum homology from amino acid residues 439–467, 715–816, and 829–910 which could be used for designing degenerate primers or probes specific for pectate lyases. The motif analysis revealed a conserved Pec_Lyase_C domain uniformly observed in all pectate lyases irrespective of variable sources suggesting its possible role in structural and enzymatic functions.

  4. Azospirillum irakense Produces a Novel Type of Pectate Lyase

    Science.gov (United States)

    Bekri, My Ali; Desair, Jos; Keijers, Veerle; Proost, Paul; Searle-van Leeuwen, Marjo; Vanderleyden, Jos; vande Broek, Ann

    1999-01-01

    The pelA gene from the N2-fixing plant-associated bacterium Azospirillum irakense, encoding a pectate lyase, was isolated by heterologous expression in Escherichia coli. Nucleotide sequence analysis of the region containing pelA indicated an open reading frame of 1,296 bp, coding for a preprotein of 432 amino acids with a typical amino-terminal signal peptide of 24 amino acids. N-terminal amino acid sequencing confirmed the processing of the protein in E. coli at the signal peptidase cleavage site predicted by nucleotide sequence analysis. Analysis of the amino acid sequence of PelA revealed no homology to other known pectinases, indicating that PelA belongs to a new pectate lyase family. PelA macerates potato tuber tissue, has an alkaline pH optimum, and requires Ca2+ for its activity. Of several divalent cations tested, none could substitute for Ca2+. Methyl-esterified pectin (with a degree of esterification up to 93%) and polygalacturonate can be used as substrates. Characterization of the degradation products formed upon incubation with polygalacturonate indicated that PelA is an endo-pectate lyase generating unsaturated digalacturonide as the major end product. Regulation of pelA expression was studied by means of a translational pelA-gusA fusion. Transcription of this fusion is low under all growth conditions tested and is dependent on the growth phase. In addition, pelA expression was found to be induced by pectin. An A. irakense pelA::Tn5 mutant still displayed pectate lyase activity, suggesting the presence of multiple pectate lyase genes in A. irakense. PMID:10198006

  5. Azospirillum irakense produces a novel type of pectate lyase.

    Science.gov (United States)

    Bekri, M A; Desair, J; Keijers, V; Proost, P; Searle-van Leeuwen, M; Vanderleyden, J; Vande Broek, A

    1999-04-01

    The pelA gene from the N2-fixing plant-associated bacterium Azospirillum irakense, encoding a pectate lyase, was isolated by heterologous expression in Escherichia coli. Nucleotide sequence analysis of the region containing pelA indicated an open reading frame of 1,296 bp, coding for a preprotein of 432 amino acids with a typical amino-terminal signal peptide of 24 amino acids. N-terminal amino acid sequencing confirmed the processing of the protein in E. coli at the signal peptidase cleavage site predicted by nucleotide sequence analysis. Analysis of the amino acid sequence of PelA revealed no homology to other known pectinases, indicating that PelA belongs to a new pectate lyase family. PelA macerates potato tuber tissue, has an alkaline pH optimum, and requires Ca2+ for its activity. Of several divalent cations tested, none could substitute for Ca2+. Methyl-esterified pectin (with a degree of esterification up to 93%) and polygalacturonate can be used as substrates. Characterization of the degradation products formed upon incubation with polygalacturonate indicated that PelA is an endo-pectate lyase generating unsaturated digalacturonide as the major end product. Regulation of pelA expression was studied by means of a translational pelA-gusA fusion. Transcription of this fusion is low under all growth conditions tested and is dependent on the growth phase. In addition, pelA expression was found to be induced by pectin. An A. irakense pelA::Tn5 mutant still displayed pectate lyase activity, suggesting the presence of multiple pectate lyase genes in A. irakense.

  6. Ammonia diffusion through Nalophan™ bags.

    Science.gov (United States)

    Sironi, Selena; Eusebio, Lidia; Dentoni, Licinia; Capelli, Laura; Del Rosso, Renato

    2014-01-01

    The aim of the work is to verify the diffusion rate of ammonia through the Nalophan™ film that constitutes the sampling bag, considering storage times ranging from 1 to 26 h. The ammonia decay over time was evaluated using gas-chromatography for the quantification of ammonia concentration inside the bag. The research assesses the roles of both of ammonia and water concentration gradients at the polymeric film interface on the diffusion process. The results show that both the ammonia concentration gradient and, in a less pronounced way, the water concentration gradient are the main 'engines' of ammonia diffusion. Double bags seem to represent a simple solution for preventing ammonia losses during storage. Another interesting result concerns the role of the bag surface on the ammonia diffusion rate: the higher the surface/volume (S/V) ratio, the higher the ammonia diffusion rate through the polymeric film.

  7. Adenylosuccinate lyase deficiency in a Malaysian patient, with novel adenylosuccinate lyase gene mutations.

    Science.gov (United States)

    Chen, Bee Chin; McGown, Ivan N; Thong, Meow Keong; Pitt, James; Yunus, Zabedah M; Khoo, Teck Beng; Ngu, Lock Hock; Duley, John A

    2010-12-01

    Most cases of adenylosuccinate lyase (ADSL OMIM 103050) deficiency reported to date are confined to the various European ethnic groups. We report on the first Malaysian case of ADSL deficiency, which appears also to be the first reported Asian case. The case was diagnosed among a cohort of 450 patients with clinical features of psychomotor retardation, global developmental delay, seizures, microcephaly and/or autistic behaviour. The patient presented with frequent convulsions and severe myoclonic jerk within the first few days of life and severe psychomotor retardation. The high performance liquid chromatography (HPLC) profile of the urine revealed the characteristic biochemical markers of succinyladenosine (S-Ado) and succinyl-aminoimidazole carboximide riboside (SAICAr). The urinary S-Ado/SAICAr ratio was found to be 1.02 (type I ADSL deficiency). The patient was compound heterozygous for two novel mutations, c.445C > G (p.R149G) and c.774_778insG (p.A260GfsX24).

  8. Cultivable Alginate Lyase-Excreting Bacteria Associated with the Arctic Brown Alga Laminaria

    Directory of Open Access Journals (Sweden)

    Yu-Zhong Zhang

    2012-11-01

    Full Text Available Although some alginate lyases have been isolated from marine bacteria, alginate lyases-excreting bacteria from the Arctic alga have not yet been investigated. Here, the diversity of the bacteria associated with the brown alga Laminaria from the Arctic Ocean was investigated for the first time. Sixty five strains belonging to nine genera were recovered from six Laminaria samples, in which Psychrobacter (33/65, Psychromonas (10/65 and Polaribacter (8/65 were the predominant groups. Moreover, 21 alginate lyase-excreting strains were further screened from these Laminaria-associated bacteria. These alginate lyase-excreting strains belong to five genera. Psychromonas (8/21, Psedoalteromonas (6/21 and Polaribacter (4/21 are the predominant genera, and Psychrobacter, Winogradskyella, Psychromonas and Polaribacter were first found to produce alginate lyases. The optimal temperatures for the growth and algiante lyase production of many strains were as low as 10–20 °C, indicating that they are psychrophilic bacteria. The alginate lyases produced by 11 strains showed the highest activity at 20–30 °C, indicating that these enzymes are cold-adapted enzymes. Some strians showed high levels of extracellular alginate lyase activity around 200 U/mL. These results suggest that these algiante lyase-excreting bacteria from the Arctic alga are good materials for studying bacterial cold-adapted alginate lyases.

  9. Cultivable alginate lyase-excreting bacteria associated with the Arctic brown alga Laminaria.

    Science.gov (United States)

    Dong, Sheng; Yang, Jie; Zhang, Xi-Ying; Shi, Mei; Song, Xiao-Yan; Chen, Xiu-Lan; Zhang, Yu-Zhong

    2012-11-06

    Although some alginate lyases have been isolated from marine bacteria, alginate lyases-excreting bacteria from the Arctic alga have not yet been investigated. Here, the diversity of the bacteria associated with the brown alga Laminaria from the Arctic Ocean was investigated for the first time. Sixty five strains belonging to nine genera were recovered from six Laminaria samples, in which Psychrobacter (33/65), Psychromonas (10/65) and Polaribacter (8/65) were the predominant groups. Moreover, 21 alginate lyase-excreting strains were further screened from these Laminaria-associated bacteria. These alginate lyase-excreting strains belong to five genera. Psychromonas (8/21), Psedoalteromonas (6/21) and Polaribacter (4/21) are the predominant genera, and Psychrobacter, Winogradskyella, Psychromonas and Polaribacter were first found to produce alginate lyases. The optimal temperatures for the growth and algiante lyase production of many strains were as low as 10–20 °C, indicating that they are psychrophilic bacteria. The alginate lyases produced by 11 strains showed the highest activity at 20–30 °C, indicating that these enzymes are cold-adapted enzymes. Some strians showed high levels of extracellular alginate lyase activity around 200 U/mL. These results suggest that these algiante lyase-excreting bacteria from the Arctic alga are good materials for studying bacterial cold-adapted alginate lyases.

  10. Tritiated ammonia formation

    International Nuclear Information System (INIS)

    Heung, L.K.

    1995-01-01

    When nitrogen was selected as the glovebox atmosphere for the Replacement Tritium Facility (RTF) at the Savannah River Site (SRS), a concern was raised as to the possibility of tritiated ammonia formation in the gloveboxes. Experimental data were produced to study the tritiated ammonia formation rate in a tritium and nitrogen mixture. A rate equation that closely simulates the experimental data was developed. This rate equation can be used to calculate the formation of tritiated ammonia from different concentrations of tritium and nitrogen. The reaction of T 2 and N 2 to form NT 3 is a slow process, particularly when the tritium concentration is low. The reaction requires weeks or months to reach radiochemical equilibrium dependent on the concentrations of the reactants. 4 refs., 6 figs., 1 tab

  11. Respiratory ammonia output and blood ammonia concentration during incremental exercise

    NARCIS (Netherlands)

    Ament, W; Huizenga, [No Value; Kort, E; van der Mark, TW; Grevink, RG; Verkerke, GJ

    The aim of this study was to investigate whether the increase of ammonia concentration and lactate concentration in blood was accompanied by an increased expiration of ammonia during graded exercise. Eleven healthy subjects performed an incremental cycle ergometer test. Blood ammonia, blood lactate

  12. Liquid ammonia injury.

    Science.gov (United States)

    George, A; Bang, R L; Lari, A R; Gang, R K; Kanjoor, J R

    2000-06-01

    The toxic effects of a gas depend on the time of exposure, concentration and its chemical nature. Pressurized liquids and gases exert an additional cold thermal injury and this may complicate the clinical picture. A patient who had an accidental exposure to liquid ammonia over a prolonged period, manifesting in cutaneous, respiratory and ocular damage in addition to a severe cold thermal injury (frostbite) with a fatal outcome is presented. The patient had flaccid quadriparesis and episodes of bradycardia, which has not been reported previously. These manifestations raise the possibility of the systemic toxicity in patients with prolonged exposure to ammonia.

  13. Ammonia emissions in Europe

    DEFF Research Database (Denmark)

    Jacobsen, Brian H.

    2012-01-01

    The NEC (National Emission Ceiling) directive has set targets for the 2010 ammonia emissions from a number of European countries. The target will be reached by most EU-countries and the total emission for EU-27 has been reduced by 22% from 1990 to 2007. Denmark is one of the countries with the la......The NEC (National Emission Ceiling) directive has set targets for the 2010 ammonia emissions from a number of European countries. The target will be reached by most EU-countries and the total emission for EU-27 has been reduced by 22% from 1990 to 2007. Denmark is one of the countries...

  14. The Ammonia-Soda Process.

    Science.gov (United States)

    Tingle, M.

    1979-01-01

    This article is a condensed version of a commentary written to accompany a set of slides which describes the ammonia-soda process used by the ammonia-soda plant at Northwich of the United Kingdom. (HM)

  15. Liberation of ammonia by cyanobacteria

    Energy Technology Data Exchange (ETDEWEB)

    Newton, J.W.

    1986-04-01

    Photoheterotrophic nitrogen-fixing cyanobacteria release ammonia when treated with methionine sulfoximine (MSX) to inhibit nitrogen incorporation into protein. This released ammonia can be derived from recently fixed nitrogen (nitrogen atmosphere) or endogenous reserves (argon atmosphere). Anaerobic ammonia release requires light and is stimulated by the photosystem II herbicides DCMU and Atrazine, regardless of the source of ammonia. As much as one quarter of the total cellular nitrogen can be released as ammonia by cyanbacteria treated with MSX and DCMU under argon in light. Chromatography of cell extracts indicates that virtually all cellular proteins are degraded. DCMU and Atrazine, at very low concentration, inhibit sustained uptake of the ammonia analog /sup 14/C methylamine. These data indicate that the herbicides interrupt ammonia uptake and retention by the cells, and support a role for photosystem II in ammonia metabolism.

  16. Crystallization and preliminary X-ray analysis of an exotype alginate lyase Atu3025 from Agrobacterium tumefaciens strain C58, a member of polysaccharide lyase family 15

    Energy Technology Data Exchange (ETDEWEB)

    Ochiai, Akihito [Laboratory of Basic and Applied Molecular Biotechnology, Graduate School of Agriculture, Kyoto University, Uji, Kyoto 611-0011 (Japan); Yamasaki, Masayuki; Mikami, Bunzo [Laboratory of Food Quality Design and Development, Graduate School of Agriculture, Kyoto University, Uji, Kyoto 611-0011 (Japan); Hashimoto, Wataru; Murata, Kousaku, E-mail: kmurata@kais.kyoto-u.ac.jp [Laboratory of Basic and Applied Molecular Biotechnology, Graduate School of Agriculture, Kyoto University, Uji, Kyoto 611-0011 (Japan)

    2006-05-01

    The crystallization and preliminary X-ray characterization of a family PL-15 exotype alginate lyase are presented. Almost all alginate lyases depolymerize alginate in an endolytical fashion via a β-elimination reaction. The alginate lyase Atu3025 from Agrobacterium tumefaciens strain C58, consisting of 776 amino-acid residues, is a novel exotype alginate lyase classified into polysaccharide lyase family 15. The enzyme was crystallized at 293 K by sitting-drop vapour diffusion with polyethylene glycol 4000 as a precipitant. Preliminary X-ray analysis showed that the Atu3025 crystal belonged to space group P2{sub 1} and diffracted to 2.8 Å resolution, with unit-cell parameters a = 107.7, b = 108.3, c = 149.5 Å, β = 91.5°.

  17. Crystallization and preliminary X-ray analysis of an exotype alginate lyase Atu3025 from Agrobacterium tumefaciens strain C58, a member of polysaccharide lyase family 15

    International Nuclear Information System (INIS)

    Ochiai, Akihito; Yamasaki, Masayuki; Mikami, Bunzo; Hashimoto, Wataru; Murata, Kousaku

    2006-01-01

    The crystallization and preliminary X-ray characterization of a family PL-15 exotype alginate lyase are presented. Almost all alginate lyases depolymerize alginate in an endolytical fashion via a β-elimination reaction. The alginate lyase Atu3025 from Agrobacterium tumefaciens strain C58, consisting of 776 amino-acid residues, is a novel exotype alginate lyase classified into polysaccharide lyase family 15. The enzyme was crystallized at 293 K by sitting-drop vapour diffusion with polyethylene glycol 4000 as a precipitant. Preliminary X-ray analysis showed that the Atu3025 crystal belonged to space group P2 1 and diffracted to 2.8 Å resolution, with unit-cell parameters a = 107.7, b = 108.3, c = 149.5 Å, β = 91.5°

  18. Quantitation of heparosan with heparin lyase III and spectrophotometry.

    Science.gov (United States)

    Huang, Haichan; Zhao, Yingying; Lv, Shencong; Zhong, Weihong; Zhang, Fuming; Linhardt, Robert J

    2014-02-15

    Heparosan is Escherichia coli K5 capsule polysaccharide, which is the key precursor for preparing bioengineered heparin. A rapid and effective quantitative method for detecting heparosan is important in the large-scale production of heparosan. Heparin lyase III (Hep III) effectively catalyzes the heparosan depolymerization, forming unsaturated disaccharides that are measurable using a spectrophotometer at 232 nm. We report a new method for the quantitative detection of heparosan with heparin lyase III and spectrophotometry that is safer and more specific than the traditional carbazole assay. In an optimized detection system, heparosan at a minimum concentration of 0.60 g/L in fermentation broth can be detected. Copyright © 2013 Elsevier Inc. All rights reserved.

  19. Isolation of protoplasts from undaria pinnatifida by alginate lyase digestion

    Science.gov (United States)

    Xiaoke, Hu; Xiaolu, Jiang; Huashi, Guan

    2003-04-01

    The aim of this study is to isolate protoplasts from Undaria pinnatifida. Protoplasts of the alga were isolated enzymatically by using alginate lyase, which was prepared by fermenting culture of a strain Vibrio sp. 510. Monofacterial method was applied for optimizing digestion condition. The optimum condition for protoplast preparation is enzymatic digestion at 28°C for 2h using alginate lyase at the concentration of 213.36 U (8 mL) every 0.5g fresh thalline with NaCl 50 and at the shaking speed of 150 r min-1 during digestion. The protoplast yield can reach 2.62±0.09 million per 0.5 g fresh leave under the optimum condition. The enzyme activity is inhibited by Ca2+ and slightly enhanced by Fe2+ and Mn2+ at concentrations of 0.05, 0.08 and 0.10 mol L-1.

  20. Function analysis of cystathionine gamma-lyase mutants

    Czech Academy of Sciences Publication Activity Database

    Hašek, Jindřich

    2009-01-01

    Roč. 16, 1a (2009), b5 ISSN 1211-5894. [Discussions in Structural Molecular Biology /7./. 12.03.2009-14.03.2009, Nové Hrady] R&D Projects: GA ČR GA305/07/1073 Institutional research plan: CEZ:AV0Z40500505 Keywords : cystathionine gamma-lyase * medical treatment * vitamin B6 Subject RIV: CD - Macromolecular Chemistry

  1. Similarities in the action patterns of exopolygalacturonate lyase and pectinesterase from Clostridium multifermentans.

    Science.gov (United States)

    Lee, M; Miller, L; Macmillan, J D

    1970-09-01

    Exopolygalacturonate lyase and pectinesterase from Clostridium multifermentans were assayed simultaneously in the same reaction mixture which contained a highly esterified pectin, polymethyl polygalacturonic acid methyl glycoside. Lyase is specific for unesterified galacturonide residues and cannot degrade this substrate in the absence of the esterase. The rate for esterase was twice the rate for lyase throughout the entire course of the combined reaction. Thus, the molar ratio of the two enzyme activities was the same since the product of the lyase is an unsaturated digalacturonic acid containing two free carboxyl groups. Since clostridial exopolygalacturonate lyase is known to degrade polygalacturonate in a linear manner beginning from the reducing ends of polygalacturonate chains, it was apparent that clostridial pectinesterase must hydrolyze methyl groups in highly esterified pectins with an action pattern similar to that of the lyase. Otherwise it would be impossible for the two enzyme rates to have corresponded on the basis of a 2:1 ratio.

  2. Hydroaminomethylation in supercritical ammonia

    Energy Technology Data Exchange (ETDEWEB)

    Martin, A.; Kant, M. [Leibniz-Institute for Catalysis, Berlin (Germany); Klein, H.; Jackstell, R.; Beller, M. [Leibniz-Institute for Catalysis, Rostock (Germany)

    2006-07-01

    Thermodynamic measurements were carried in the reaction system of hydroaminomethylation of olefins. Mixtures of ammonia, olefins, co-solvents, syngas and products such as nonylamine used as model and water were studied. In dependence on the reaction conditions and the mixtures selected opalescence points in a region from 92-290 bar and 120-172 C were found. (orig.)

  3. Ammonia abundances in comets

    Science.gov (United States)

    Wyckoff, S.; Tegler, S.; Engel, L.

    The emission band strengths of the NH2 bands of Comets Halley, Hartley-Good, Thiele, and Borrelly were measured to determine the NH2 column densities for the comets. Production rates obtained using the Haser and vectorial models are in agreement within the observational errors, suggesting that a simple two-step decay model may be used to approximate the NH2 distribution in a comet's coma. Ammonia-to-water abundance ratios from 0.01 to 0.4 percent were found for the four comets. The ratio in Comet Halley is found to be Q(NH3)/Q(H2O) = 0.002 + or - 0.001. No significant difference in the ammonia abundance was found before or after perihelion in Comet Halley.

  4. Alfalfa contains substantial 9-hydroperoxide lyase activity and a 3Z:2E-enal isomerase

    NARCIS (Netherlands)

    Vliegenthart, J.F.G.; Noordermeer, M.A.; Veldink, G.A.

    1999-01-01

    Fatty acid hydroperoxides formed by lipoxygenase can be cleaved by hydroperoxide lyase resulting in the formation of short-chain aldehydes and omega-oxo acids. Plant hydroperoxide lyases use 13- or 9-hydroperoxy linoleic and linolenic acid as substrates. Alfalfa (Medicago sativa L.) has been

  5. Overexpression of isocitrate lyase-glyoxylate bypass influence on metabolism in Aspergillus niger

    DEFF Research Database (Denmark)

    Meijer, Susan Lisette; Otero, José Manuel; Olivares Hernandez, Roberto

    2009-01-01

    In order to improve the production of succinate and malate by the filamentous fungus Aspergillus niger the activity of the glyoxylate bypass pathway was increased by over-expression of the isocitrate lyase (icl) gene. The hypothesis was that when isocitrate lyase was up-regulated the flux towards...

  6. The PduQ enzyme is an alcohol dehydrogenase used to recycle NAD+ internally within the Pdu microcompartment of Salmonella enterica.

    Directory of Open Access Journals (Sweden)

    Shouqiang Cheng

    Full Text Available Salmonella enterica uses a bacterial microcompartment (MCP for coenzyme B(12-dependent 1,2-propanediol (1,2-PD utilization (Pdu. The Pdu MCP consists of a protein shell that encapsulates enzymes and cofactors required for metabolizing 1,2-PD as a carbon and energy source. Here we show that the PduQ protein of S. enterica is an iron-dependent alcohol dehydrogenase used for 1,2-PD catabolism. PduQ is also demonstrated to be a new component of the Pdu MCP. In addition, a series of in vivo and in vitro studies show that a primary function of PduQ is to recycle NADH to NAD(+ internally within the Pdu MCP in order to supply propionaldehyde dehydrogenase (PduP with its required cofactor (NAD(+. Genetic tests determined that a pduQ deletion mutant grew slower than wild-type Salmonella on 1,2-PD and that this phenotype was not complemented by a non-MCP associated Adh2 from Zymomonas that catalyzes the same reaction. This suggests that PduQ has a MCP-specific function. We also found that a pduQ deletion mutant had no growth defect in a genetic background having a second mutation that prevents MCP formation which further supports a MCP-specific role for PduQ. Moreover, studies with purified Pdu MCPs demonstrated that the PduQ enzyme can convert NADH to NAD(+ to supply the PduP reaction in vitro. Cumulatively, these studies show that the PduQ enzyme is used to recycle NADH to NAD(+ internally within the Pdu MCP. To our knowledge, this is the first report of internal recycling as a mechanism for cofactor homeostasis within a bacterial MCP.

  7. Structural insights into the bacterial carbon - phosphorus lyase machinery

    DEFF Research Database (Denmark)

    Seweryn, Paulina; Van, Lan Bich; Kjeldgaard, Morten

    2015-01-01

    Phosphorus is required for all life and microorganisms can extract it from their environment through several metabolic pathways. When phosphate is in limited supply, some bacteria are able to use phosphonate compounds, which require specialized enzymatic machinery to break the stable carbon–phosphorus...... (C–P) bond. Despite its importance, the details of how this machinery catabolizes phosphonates remain unknown. Here we determine the crystal structure of the 240-kilodalton Escherichia coli C–P lyase core complex (PhnG–PhnH–PhnI–PhnJ; PhnGHIJ), and show that it is a two-fold symmetric hetero...

  8. Hydroxynitrile Lyases: Biological Sources and Application as Biocatalysts

    Directory of Open Access Journals (Sweden)

    Herfried Griengl

    2004-01-01

    Full Text Available We review the state of the art regarding the application of hydroxynitrile lyases to obtain, enantioselectively, (R- and (S-cyanohydrins of aldehydes and ketones. Special emphasis is given to recent preparative applications and to research for extending the number of plants serving as sources for the enzyme. Depending on the plant family, the mechanism of the enzyme-catalysed reaction can be different. A novel area of research is the consideration of evolutionary aspects on the basis of structure comparisons.

  9. Adenylosuccinate lyase deficiency: the first identified polish patient.

    Science.gov (United States)

    Jurkiewicz, Elzbieta; Mierzewska, Hanna; Kuśmierska, Katarzyna

    2007-10-01

    Adenylosuccinate lyase (ADSL) deficiency is a rare disease of de novo purine synthesis. The main symptoms are psychomotor retardation, epilepsy, autistic features, occasionally associated with muscular hypotonia. Diagnosis is made by detection of abnormal purine metabolites (succinyladenosine - S-Ado and succinylaminoimidazole carboxamide riboside - SAICAr) in body fluids. The severity of the clinical features correlates with low S-Ado/SAICAr ratio. We report clinical, biochemical and brain MRI findings of a female infant with severe early epilepsy and hypotonia, who died at the age of 10 weeks.

  10. Azospirillum irakense Produces a Novel Type of Pectate Lyase

    OpenAIRE

    Bekri, My Ali; Desair, Jos; Keijers, Veerle; Proost, Paul; Searle-van Leeuwen, Marjo; Vanderleyden, Jos; vande Broek, Ann

    1999-01-01

    The pelA gene from the N2-fixing plant-associated bacterium Azospirillum irakense, encoding a pectate lyase, was isolated by heterologous expression in Escherichia coli. Nucleotide sequence analysis of the region containing pelA indicated an open reading frame of 1,296 bp, coding for a preprotein of 432 amino acids with a typical amino-terminal signal peptide of 24 amino acids. N-terminal amino acid sequencing confirmed the processing of the protein in E. coli at the signal peptidase cleavage...

  11. Design of thermostable rhamnogalacturonan lyase mutants from Bacillus licheniformis by combination of targeted single point mutations

    DEFF Research Database (Denmark)

    da Silva, Ines Isabel Cardoso Rodrigues; Jers, Carsten; Otten, Harm

    2014-01-01

    Rhamnogalacturonan I lyases (RGI lyases) (EC 4.2.2.-) catalyze cleavage of α-1,4 bonds between rhamnose and galacturonic acid in the backbone of pectins by β-elimination. In the present study, targeted improvement of the thermostability of a PL family 11 RGI lyase from Bacillus licheniformis (DSM...... the wild-type RGI lyase in Bacillus subtilis as opposed to in Pichia pastoris; this effect is suggested to be a negative result of glycosylation of the P. pastoris expressed enzyme. A ~ twofold improvement in thermal stability at 60 °C, accompanied by less significant increases in Tm of the enzyme mutants......, were obtained due to additive stabilizing effects of single amino acid mutations (E434L, G55V, and G326E) compared to the wild type. The crystal structure of the B. licheniformis wild-type RGI lyase was also determined; the structural analysis corroborated that especially mutation of charged amino...

  12. Elimination of hydrogen sulphide and β substitution in cystein, catalyzed by the cysteine-lyase of hens yolk-sac and yolk (1961)

    International Nuclear Information System (INIS)

    Chapeville, F.; Fromageot, P.

    1961-01-01

    The yolk of incubated hen's eggs contains a pyridoxal phosphate activated enzyme, free of iron, copper, magnesium and calcium. This enzyme activates the β-carbon atom of cysteine. Its reactivity is demonstrated by the ease with which this β-carbon fixes various sulfur containing substances in which the sulfur has reducing properties: inorganic sulfide, sulfide or cysteine itself. In the absence of substances able to react with the β-carbon atom, the active complex, consisting of the enzyme and the aminated tri-carbon chain, is hydrolysed to pyruvic acid and ammonia. The liberation of hydrogen sulfide thus appears to be the consequence either of the substitution of the β-carbon atom of cysteine or of the decomposition of the complex which this aminoacid forms with the enzyme studied. The latter seems therefore to possess an activity which differs from the activity of the desulfhydrases as yet known. We suggest to call this enzyme cystein-lyase. (authors) [fr

  13. Screening of alginate lyase-excreting microorganisms from the surface of brown algae.

    Science.gov (United States)

    Wang, Mingpeng; Chen, Lei; Zhang, Zhaojie; Wang, Xuejiang; Qin, Song; Yan, Peisheng

    2017-12-01

    Alginate lyase is a biocatalyst that degrades alginate to produce oligosaccharides, which have many bioactive functions and could be used as renewable biofuels. Here we report a simple and sensitive plate assay for screening alginate lyase-excreting microorganisms from brown algae. Brown algae Laminaria japonica, Sargassum horneri and Sargassum siliquatrum were cultured in sterile water. Bacteria growing on the surface of seaweeds were identified and their capacity of excreting alginate lyase was analyzed. A total of 196 strains were recovered from the three different algae samples and 12 different bacterial strains were identified capable of excreting alginate lyases. Sequence analysis of the 16S rRNA gene revealed that these alginate lyase-excreting strains belong to eight genera: Paenibacillus (4/12), Bacillus (2/12), Leclercia (1/12), Isoptericola (1/12), Planomicrobium (1/12), Pseudomonas (1/12), Lysinibacillus (1/12) and Sphingomonas (1/12). Further analysis showed that the LJ-3 strain (Bacillus halosaccharovorans) had the highest enzyme activity. To our best knowledge, this is the first report regarding alginate lyase-excreting strains in Paenibacillus, Planomicrobium and Leclercia. We believe that our method used in this study is relatively easy and reliable for large-scale screening of alginate lyase-excreting microorganisms.

  14. Renal Ammonia Metabolism and Transport

    Science.gov (United States)

    Weiner, I. David; Verlander, Jill W.

    2015-01-01

    Renal ammonia metabolism and transport mediates a central role in acid-base homeostasis. In contrast to most renal solutes, the majority of renal ammonia excretion derives from intrarenal production, not from glomerular filtration. Renal ammoniagenesis predominantly results from glutamine metabolism, which produces 2 NH4+ and 2 HCO3− for each glutamine metabolized. The proximal tubule is the primary site for ammoniagenesis, but there is evidence for ammoniagenesis by most renal epithelial cells. Ammonia produced in the kidney is either excreted into the urine or returned to the systemic circulation through the renal veins. Ammonia excreted in the urine promotes acid excretion; ammonia returned to the systemic circulation is metabolized in the liver in a HCO3−-consuming process, resulting in no net benefit to acid-base homeostasis. Highly regulated ammonia transport by renal epithelial cells determines the proportion of ammonia excreted in the urine versus returned to the systemic circulation. The traditional paradigm of ammonia transport involving passive NH3 diffusion, protonation in the lumen and NH4+ trapping due to an inability to cross plasma membranes is being replaced by the recognition of limited plasma membrane NH3 permeability in combination with the presence of specific NH3-transporting and NH4+-transporting proteins in specific renal epithelial cells. Ammonia production and transport are regulated by a variety of factors, including extracellular pH and K+, and by several hormones, such as mineralocorticoids, glucocorticoids and angiotensin II. This coordinated process of regulated ammonia production and transport is critical for the effective maintenance of acid-base homeostasis. PMID:23720285

  15. CADDIS Volume 2. Sources, Stressors and Responses: Ammonia

    Science.gov (United States)

    Introduction to the ammonia module, when to list ammonia as a candidate cause, ways to measure ammonia, simple and detailed conceptual diagrams for ammonia, literature reviews and references for the ammonia module.

  16. Sequence analysis and overexpression of a pectin lyase gene (pel1) from Aspergillus oryzae KBN616.

    Science.gov (United States)

    Kitamoto, N; Yoshino-Yasuda, S; Ohmiya, K; Tsukagoshi, N

    2001-01-01

    A gene (pel1) encoding pectin lyase (Pel1) was isolated from a shoyu koji mold, Aspergillus oryzae KBN616, and characterized. The structural gene comprised 1,196 bp with a single intron. The ORF encoded 381 amino acids with a signal peptide of 20 amino acids. The deduced amino acid sequence showed high similarity to those of Aspergillus niger pectin lyases and Glomerella cingulata PnlA. The pel1 gene was successfully overexpressed under the promoter of the A. oryzae TEF1 gene. The molecular mass of the recombinant pectin lyase substantially coincided with that calculated based on nucleotide sequence.

  17. Sulfur mobilization in cyanobacteria: the catalytic mechanism of L-cystine C-S lyase (C-DES) from synechocystis.

    Science.gov (United States)

    Campanini, Barbara; Schiaretti, Francesca; Abbruzzetti, Stefania; Kessler, Dorothea; Mozzarelli, Andrea

    2006-12-15

    Sulfur mobilization represents one of the key steps in ubiquitous Fe-S clusters assembly and is performed by a recently characterized set of proteins encompassing cysteine desulfurases, assembly factors, and shuttle proteins. Despite the evolutionary conservation of these proteins, some degree of variability among organisms was observed, which might reflect functional specialization. L-Cyst(e)ine lyase (C-DES), a pyridoxal 5'-phosphatedependent enzyme identified in the cyanobacterium Synechocystis, was reported to use preferentially cystine over cysteine with production of cysteine persulfide, pyruvate, and ammonia. In this study, we demonstrate that C-DES sequences are present in all cyanobacterial genomes and constitute a new family of sulfur-mobilizing enzymes, distinct from cysteine desulfurases. The functional properties of C-DES from Synechocystis sp. PCC 6714 were investigated under pre-steady-state and steady-state conditions. Single wavelength and rapid scanning stopped-flow kinetic data indicate that the internal aldimine reacts with cystine forming an external aldimine that rapidly decays to a transient quinonoid species and stable tautomers of the alpha-aminoacrylate Schiff base. In the presence of cysteine, the transient formation of a dipolar species precedes the selective and stable accumulation of the enolimine tautomer of the external aldimine, with no formation of the alpha-aminoacrylate Schiff base under reducing conditions. Effective sulfur mobilization from cystine might represent a mechanism that allows adaptation of cyanobacteria to different environmental conditions and to light-dark cycles.

  18. Structural Insights Into The Bacterial Carbon-Phosphorus Lyase Machinery

    DEFF Research Database (Denmark)

    Brodersen, Ditlev Egeskov

    the proteins encoded in the phn operon act in concert to catabolise phosphonate remain unknown. We have determined the crystal structure of a 240 kDa Escherichia coli carbon-phosphorus lyase core complex at 1.7 Å and show that it comprises a highly intertwined network of subunits with several unexpected......Phosphonate compounds act as a nutrient source for some microorganisms when phosphate is limiting but require a specialised enzymatic machinery due to the presence of the highly stable carbon-phosphorus bond. Despite the fundamental importance to microbial metabolism, the details of how...... structural features. The complex contains at least two different active sites and suggest a revision of current models of carbon-phosphorus bond cleavage. Using electron microscopy, we map the binding site of an additional protein subunit, which may use ATP for driving conformational changes during...

  19. Anhydrous Ammonia Frost on Titan

    Science.gov (United States)

    Smythe, W. D.; Nelson, R.; Boryta, M. D.

    2009-12-01

    Ammonia has been suggested as a probable source for sustaining Titan's thick nitrogen-dominated atmosphere. Ammonia is believed to be important to maintaining nitrogen in Titan's atmosphere. Ammonia is seen in clouds in the atmospheres of Jupiter and Saturn, but has yet to be detected on any of the satellites. This may be because all forms of NH3 are unstable in the ambient conditions of the satellites surfaces or that its spectral features are altered by other components of the surface, and have not been identified. It has recently been demonstrated[1] that brightening occurs in Titan’s atmosphere that is transient on the time-scale of months. The spectral shape of the brightening is more consistent with that of the transient apparition of a pure ammonia frost than of an ammonia monohydrate or ammonia dihydrate frost. However, the phase behavior of the ammonia water system has peritectics at compositions of 1:1 and 1:2. These hydrate forms would be expected to dominate if the frost, or the reservoir from which the frost was derived had any water present. Physical mechanisms for producing measurable quanitities of anhydrous ammonia can include chemical dehydration or dehydration of the vapor phase - but it is challenging to store significant quantities of the anhydrous material because of the phase behavior in the solid state. [1] Nelson, R.M., et al. Saturn’s Titan: Surface Change, Ammonia, and Implications for Atmospheric and Tectonic Activity., Icarus, 199, pp. 429-441, 2009 This work was performed at JPL under contract to NASA

  20. Ammonia-based quantum computer

    International Nuclear Information System (INIS)

    Ferguson, Andrew J.; Cain, Paul A.; Williams, David A.; Briggs, G. Andrew D.

    2002-01-01

    We propose a scheme for quantum computation using two eigenstates of ammonia or similar molecules. Individual ammonia molecules are confined inside fullerenes and used as two-level qubit systems. Interaction between these ammonia qubits takes place via the electric dipole moments, and in particular we show how a controlled-NOT gate could be implemented. After computation the qubit is measured with a single-electron electrometer sensitive enough to differentiate between the dipole moments of different states. We also discuss a possible implementation based on a quantum cellular automaton

  1. Wastewater Treatment with Ammonia Recovery System

    OpenAIRE

    M. Örvös; T. Balázs; K. F. Both

    2008-01-01

    From environmental aspect purification of ammonia containing wastewater is expected. High efficiency ammonia desorption can be done from the water by air on proper temperature. After the desorption process, ammonia can be recovered and used in another technology. The calculation method described below give some methods to find either the minimum column height or ammonia rich solution of the effluent.

  2. Cytochrome c and c1 heme lyases are essential in Plasmodium berghei.

    Science.gov (United States)

    Posayapisit, Navaporn; Songsungthong, Warangkhana; Koonyosying, Pongpisid; Falade, Mofolusho O; Uthaipibull, Chairat; Yuthavong, Yongyuth; Shaw, Philip J; Kamchonwongpaisan, Sumalee

    Malaria parasites possess a de novo heme synthetic pathway. Interestingly, this pathway is dispensable during the blood stages of development in mammalian hosts. The assembly of the two most important hemeproteins, cytochromes c and c1, is mediated by cytochrome heme lyase enzymes. Plasmodium spp. possess two cytochrome heme lyases encoded by separate genes. Given the redundancy of heme synthesis, we sought to determine if heme lyase function also exhibits redundancy. To answer this question, we performed gene knockout experiments. We found that the PBANKA_143950 and PBANKA_0602600 Plasmodium berghei genes encoding cytochrome c (Pbcchl) and cytochrome c1 (Pbcc 1 hl) heme lyases, respectively, can only be disrupted when a complementary gene is present. In contrast, four genes in the de novo heme synthesis pathway can be disrupted without complementation. This work provides evidence that Pbcchl and Pbcc 1 hl are both essential and thus may be antimalarial targets. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Isocitrate lyase and the glyoxylate cycle. Progress report, February 15, 1989--February 15, 1990

    Energy Technology Data Exchange (ETDEWEB)

    McFadden, B.A.

    1990-12-31

    Active site modifications of isocitrate lyase (icl) from Escherichia coli are described. In addition directed mutagenesis of icl gene are detailed aimed at varying the charge yet conserving the structure of the enzymes active site.

  4. Genetics Home Reference: 17 alpha-hydroxylase/17,20-lyase deficiency

    Science.gov (United States)

    ... breasts and pubic hair, and do not menstruate (amenorrhea). Women with partial 17α-hydroxylase/17,20-lyase ... have female internal reproductive organs, these individuals have amenorrhea and do not develop female secondary sex characteristics. ...

  5. In vivo proton MR spectroscopy findings specific for adenylosuccinate lyase deficiency.

    NARCIS (Netherlands)

    Henneke, M.; Dreha-Kulaczewski, S.; Brockmann, K.; Graaf, M. van der; Willemsen, M.H.; Engelke, U.F.H.; Dechent, P.; Heerschap, A.; Helms, G.; Wevers, R.A.; Gartner, J.

    2010-01-01

    Adenylosuccinate lyase (ADSL) deficiency is an inherited metabolic disorder affecting predominantly the central nervous system. The disease is characterized by the accumulation of succinylaminoimidazolecarboxamide riboside and succinyladenosine (S-Ado) in tissue and body fluids. Three children

  6. Haber Process for Ammonia Synthesis

    Indian Academy of Sciences (India)

    C in order to obtain even a small percentage of ammonia. For this temperature range, however, no catalyst was available. By increasing the pressure to 75 bar the equilibrium conditions improved, but even at this pressure, and an operating.

  7. Utilization of Aspergillus oryzae to produce pectin lyase from various agro-industrial residues

    Directory of Open Access Journals (Sweden)

    Safia Koser

    2014-07-01

    Full Text Available The present study was aimed to investigate the culture influence on pectin lyase production potential of fungal strain Aspergillus oryzae. The enzyme profile of A. oryzae showed highest activity of pectin lyase after 3rd day of incubation on lemon peel waste under solid state fermentation conditions. To induce the pectin lyase synthesis capability of A. oryzae at optimal level various culture variables including physical and nutritional parameters were optimized by adopting classical optimization technique. Therefore, through fermentation process optimization the production of pectin lyase was substantially induced up to the level of 875 U/mL, when fermentation medium of lemon peel waste inoculated with 5 mL spore suspension of A. oryzae. The optimal fermentation conditions for maximum pectin lyase yield were as: optimum pH 5, 70% moisture level and incubated at 40 °C in addition with 1% sterile glucose solution as readily available carbon source and 0.2% yeast extract as an inexpensive nitrogen supplement (1%. The results obtained in current investigation so far demonstrated that culture conditions have great influence on the pectin lyase production potential of A. oryzae.

  8. Aspergillus fumigatus Does Not Require Fatty Acid Metabolism via Isocitrate Lyase for Development of Invasive Aspergillosis▿

    Science.gov (United States)

    Schöbel, Felicitas; Ibrahim-Granet, Oumaïma; Avé, Patrick; Latgé, Jean-Paul; Brakhage, Axel A.; Brock, Matthias

    2007-01-01

    Aspergillus fumigatus is the most prevalent airborne filamentous fungus causing invasive aspergillosis in immunocompromised individuals. Only a limited number of determinants directly associated with virulence are known, and the metabolic requirements of the fungus to grow inside a host have not yet been investigated. Previous studies on pathogenic microorganisms, i.e., the bacterium Mycobacterium tuberculosis and the yeast Candida albicans, have revealed an essential role for isocitrate lyase in pathogenicity. In this study, we generated an isocitrate lyase deletion strain to test whether this strain shows attenuation in virulence. Results have revealed that isocitrate lyase from A. fumigatus is not required for the development of invasive aspergillosis. In a murine model of invasive aspergillosis, the wild-type strain, an isocitrate lyase deletion strain, and a complemented mutant strain were similarly effective in killing mice. Moreover, thin sections demonstrated invasive growth of all strains. Additionally, thin sections of lung tissue from patients with invasive aspergillosis stained with anti-isocitrate lyase antibodies remained negative. From these results, we cannot exclude the use of lipids or fatty acids as a carbon source for A. fumigatus during invasive growth. Nevertheless, test results do imply that the glyoxylate cycle from A. fumigatus is not required for the anaplerotic synthesis of oxaloacetate under infectious conditions. Therefore, an antifungal drug inhibiting fungal isocitrate lyases, postulated to act against Candida infections, is assumed to be ineffective against A. fumigatus. PMID:17178786

  9. Coordinated action of pectinesterase and polygalacturonate lyase complex of Clostridium multifermentans.

    Science.gov (United States)

    Sheiman, M I; Macmillan, J D; Miller, L; Chase, T

    1976-05-01

    The polygalacturonate lyase and pectinesterase activities of Clostridium multifermentans, both produced extracellularly when the organism grows on pectin or polygalacturonate, have been suggested to be associated in a single complex. Both enzymic sites act on their respective substrates by single-chain action patterns, as shown by equivalent release of terminal tritium label and total product throughout the reaction. From these results, the Km and V of the lyase, and the amount of lyase activity present, we calculate the steady-state concentration of lyase substrate expected during action of the two sites on pectin if the sites are independent. No such steady-state concentration of lyase substrate was observed. Therefore, we conclude that the two types of active site act in a coordinated manner; the polysaccharide chain passes from the esterase site to the lyase site without intermediate dissociation and rebinding. This 'molecular disassembly line' constituted by the two sites may represent a system of general significance in synthesis and degradation of biological polymers.

  10. Three Alginate Lyases from Marine Bacterium Pseudomonas fluorescens HZJ216: Purification and Characterization

    Energy Technology Data Exchange (ETDEWEB)

    Liyan, Li [Ocean University of China, Qingdao, PRC; Jiang, Xiaolu [Ocean University of China, Qingdao, PRC; Wang, Peng [Ocean University of China, Qingdao, PRC; Guan, Huashi [Ocean University of China, Qingdao, PRC; Guo, Hong [ORNL

    2010-01-01

    Three alginate lyases (A, B, and C) from an alginate-degrading marine bacterium strain HZJ216 isolated from brown seaweed in the Yellow Sea of China and identified preliminarily as Pseudomonas fluorescens are purified, and their biochemical properties are described. Molecular masses of the three enzymes are determined by SDS-PAGE to be 60.25, 36, and 23 kDa with isoelectric points of 4, 4.36, and 4.59, respectively. Investigations of these enzymes at different pH and temperatures show that they are most active at pH 7.0 and 35 C. Alginate lyases A and B are stable in the pH range of 5.0 9.0, while alginate lyase C is stable in the pH range of 5.0 7.0. Among the metal ions tested, additions of Na+, K+, and Mg2+ ions can enhance the enzyme activities while Fe2+, Fe3+, Ba2+, and Zn2+ ions show inhibitory effects. The substrate specificity results demonstrate that alginate lyase C has the specificity for G block while alginate lyases A and B have the activities for both M and G blocks. It is the first report about extracellular alginate lyases with high alginate-degrading activity from P. fluorescens.

  11. Characterization of AlgMsp, an alginate lyase from Microbulbifer sp. 6532A.

    Directory of Open Access Journals (Sweden)

    Steven M Swift

    Full Text Available Alginate is a polysaccharide produced by certain seaweeds and bacteria that consists of mannuronic acid and guluronic acid residues. Seaweed alginate is used in food and industrial chemical processes, while the biosynthesis of bacterial alginate is associated with pathogenic Pseudomonas aeruginosa. Alginate lyases cleave this polysaccharide into short oligo-uronates and thus have the potential to be utilized for both industrial and medicinal applications. An alginate lyase gene, algMsp, from Microbulbifer sp. 6532A, was synthesized as an E.coli codon-optimized clone. The resulting 37 kDa recombinant protein, AlgMsp, was expressed, purified and characterized. The alginate lyase displayed highest activity at pH 8 and 0.2 M NaCl. Activity of the alginate lyase was greatest at 50°C; however the enzyme was not stable over time when incubated at 50°C. The alginate lyase was still highly active at 25°C and displayed little or no loss of activity after 24 hours at 25°C. The activity of AlgMsp was not dependent on the presence of divalent cations. Comparing activity of the lyase against polymannuronic acid and polyguluronic acid substrates showed a higher turnover rate for polymannuronic acid. However, AlgMSP exhibited greater catalytic efficiency with the polyguluronic acid substrate. Prolonged AlgMsp-mediated degradation of alginate produced dimer, trimer, tetramer, and pentamer oligo-uronates.

  12. Ammonia Synthesis at Low Pressure.

    Science.gov (United States)

    Cussler, Edward; McCormick, Alon; Reese, Michael; Malmali, Mahdi

    2017-08-23

    Ammonia can be synthesized at low pressure by the use of an ammonia selective absorbent. The process can be driven with wind energy, available locally in areas requiring ammonia for synthetic fertilizer. Such wind energy is often called "stranded," because it is only available far from population centers where it can be directly used. In the proposed low pressure process, nitrogen is made from air using pressure swing absorption, and hydrogen is produced by electrolysis of water. While these gases can react at approximately 400 °C in the presence of a promoted conventional catalyst, the conversion is often limited by the reverse reaction, which makes this reaction only feasible at high pressures. This limitation can be removed by absorption on an ammine-like calcium or magnesium chloride. Such alkaline metal halides can effectively remove ammonia, thus suppressing the equilibrium constraints of the reaction. In the proposed absorption-enhanced ammonia synthesis process, the rate of reaction may then be controlled not by the chemical kinetics nor the absorption rates, but by the rate of the recycle of unreacted gases. The results compare favorably with ammonia made from a conventional small scale Haber-Bosch process.

  13. A role for glutamate-333 of Saccharomyces cerevisiae cystathionine γ-lyase as a determinant of specificity.

    Science.gov (United States)

    Hopwood, Emily M S; Ahmed, Duale; Aitken, Susan M

    2014-02-01

    Cystathionine γ-lyase (CGL) catalyzes the hydrolysis of l-cystathionine (l-Cth), producing l-cysteine (l-Cys), α-ketobutyrate and ammonia, in the second step of the reverse transsulfuration pathway, which converts l-homocysteine (l-Hcys) to l-Cys. Site-directed variants substituting residues E48 and E333 with alanine, aspartate and glutamine were characterized to probe the roles of these acidic residues, conserved in fungal and mammalian CGL sequences, in the active-site of CGL from Saccharomyces cerevisiae (yCGL). The pH optimum of variants containing the alanine or glutamine substitutions of E333 is increased by 0.4-1.2 pH units, likely due to repositioning of the cofactor and modification of the pKa of the pyridinium nitrogen. The pH profile of yCGL-E48A/E333A resembles that of Escherichia coli cystathionine β-lyase. The effect of substituting E48, E333 or both residues is the 1.3-3, 26-58 and 124-568-fold reduction, respectively, of the catalytic efficiency of l-Cth hydrolysis. The Km(l-Cth) of E333 substitution variants is increased ~17-fold, while Km(l-OAS) is within 2.5-fold of the wild-type enzyme, indicating that residue E333 interacts with the distal amine moiety of l-Cth, which is not present in the alternative substrate O-acetyl-l-serine. The catalytic efficiency of yCGL for α,γ-elimination of O-succinyl-l-homoserine (kcat/Km(l-OSHS)=7±2), which possesses a distal carboxylate, but lacks an amino group, is 300-fold lower than that of the physiological l-Cth substrate (kcat/Km(l-Cth)=2100±100) and 260-fold higher than that of l-Hcys (kcat/Km(l-Hcys)=0.027±0.005), which lacks both distal polar moieties. The results of this study suggest that the glutamate residue at position 333 is a determinant of specificity. Copyright © 2013 Elsevier B.V. All rights reserved.

  14. Optimization of culturing condition and medium composition for the production of alginate lyase by a marine Vibrio sp. YKW-34

    Science.gov (United States)

    Fu, Xiaoting; Lin, Hong; Kim, Sang Moo

    2008-02-01

    Carbohydrases secreted by marine Vibrio sp. YKW-34 with strong Laminaria cell wall degrading ability were screened, and among them alginate lyase was found to be dominant. The effects of medium composition and culturing condition on the production of alginate lyase by marine Vibrio sp. YKW-34 in flask were investigated in this study. In the culture medium of marine broth, no alginate lyase was produced. The activity of the alginate lyase, after being induced, reached 5 UmL-1. The best inoculum volume and inoculum age were 10% and 12 h, respectively. The optimal temperature for alginate lyase production was 25°C. The fermentation medium was composed of 0.5% of Laminaria powder and 0.2% of KNO3 with an initial acidity of pH 8.0. Alginate could induce alginate lyase production but not as efficiently as Laminaria powder did. The addition of fucoidan, cellulose and glucose had negative effect on the alginate lyase production. Other kinds of nitrogen sources, such as yeast extract, beef extract and peptone, had positive effect on the growth of the microorganism and negative effect on alginate lyase production. In addition, the time course of alginate lyase production under the optimized condition was described. The optimal harvest time was 48 h.

  15. Modern Approaches to Discovering New Hydroxynitrile Lyases for Biocatalysis.

    Science.gov (United States)

    Padhi, Santosh Kumar

    2017-01-17

    Hydroxynitrile lyases (HNLs) have grown in importance from laboratory to industry due to their potential to catalyze stereoselective C-C bond-formation reactions in the synthesis of several chiral intermediates, such as enantiopure α-cyanohydrins, β-nitro alcohols, and their derivatives with multiple functional groups. With these wide applications, the demand for finding new HNLs has increased, and this has led to exploration not only of new HNLs but also of new ways to discover them. An exclusive review article on HNLs by Asano et al. in 2011 described the discovery of HNLs along with their applications. Since then many scientific advancements have been seen in this area. This article aims to highlight the modern HNL discovery approaches, based mainly on 1) genome mining, 2) use of INTMSAlign software, 3) rational design (based on a millipede HNL), 4) evolution of catalytic mechanisms, 5) protein engineering guided by catalytic mechanisms, and 6) screening of plants with cyanogen glycoside (CG) content. This description is followed by future prospects. Overall this review represents the present state and the future potential of HNL discovery approaches, and so might be hoped to be instrumental not only in exploration of new HNLs but also in the invention of methods for potential biotechnological applications. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Structural insights into the bacterial carbon-phosphorus lyase machinery.

    Science.gov (United States)

    Seweryn, Paulina; Van, Lan Bich; Kjeldgaard, Morten; Russo, Christopher J; Passmore, Lori A; Hove-Jensen, Bjarne; Jochimsen, Bjarne; Brodersen, Ditlev E

    2015-09-03

    Phosphorus is required for all life and microorganisms can extract it from their environment through several metabolic pathways. When phosphate is in limited supply, some bacteria are able to use phosphonate compounds, which require specialized enzymatic machinery to break the stable carbon-phosphorus (C-P) bond. Despite its importance, the details of how this machinery catabolizes phosphonates remain unknown. Here we determine the crystal structure of the 240-kilodalton Escherichia coli C-P lyase core complex (PhnG-PhnH-PhnI-PhnJ; PhnGHIJ), and show that it is a two-fold symmetric hetero-octamer comprising an intertwined network of subunits with unexpected self-homologies. It contains two potential active sites that probably couple phosphonate compounds to ATP and subsequently hydrolyse the C-P bond. We map the binding site of PhnK on the complex using electron microscopy, and show that it binds to a conserved insertion domain of PhnJ. Our results provide a structural basis for understanding microbial phosphonate breakdown.

  17. Characterization of Enzyme Structure-Function Relationship of Adenylosuccinate Lyase

    Science.gov (United States)

    Ray, Stephen; Patterson, David; Ghosh, Kingshuk; Wilkinson, Terry; Shaheen, Sean

    2009-10-01

    Adenylosuccinate lyase (ADSL) is an enzyme involved in de novo purine biosynthesis required for several important biological functions. Occasionally disturbances within the enzyme occur, causing a disorder known as ADSL deficiency. It is likely these mutations affect the formation of the tetramer structure by protein misfolding or aggregation. We are beginning to study fundamental properties of the enzyme structure-function relationship of Wild-Type ADSL compared to mutants associated with ADSL Deficiency with two major studies: i) Stability and formation of multimeric complexes in a heterogeneous pool of other structures, ii) Enzymatic activity and reaction kinetics studies by measuring reaction rates of the conversion of substrate into products and enzyme substrate complex formation equilibrium. Our group has successfully expressed Wild-Type (WT) and the mutants R426H and A291V in a protein expression vector and have measured their respective enzyme activity after purification. Modelling approaches for molecular interactions of monomer subunits show the trimer structure could be problematic. We have also carried out our preliminary analysis of the structure-function relationship using microscopic model for the A291V mutant compared to the WT protein.

  18. Novel proton MR spectroscopy findings in adenylosuccinate lyase deficiency.

    Science.gov (United States)

    Zulfiqar, Maria; Lin, Doris D M; Van der Graaf, Marinette; Barker, Peter B; Fahrner, Jill A; Marie, Sandrine; Morava, Eva; De Boer, Lonneke; Willemsen, Michel A A P; Vining, Eileen; Horská, Alena; Engelke, Udo; Wevers, Ron A; Maegawa, Gustavo H B

    2013-04-01

    Adenylosuccinate lyase (ADSL) deficiency is a rare inborn error of metabolism resulting in accumulation of metabolites including succinylaminoimidazole carboxamide riboside (SAICAr) and succinyladenosine (S-Ado) in the brain and other tissues. Patients with ADSL have progressive psychomotor retardation, neonatal seizures, global developmental delay, hypotonia, and autistic features, although variable clinical manifestations may make the initial diagnosis challenging. Two cases of the severe form of the disease are reported here: an 18-month-old boy with global developmental delay, intractable neonatal seizures, progressive cerebral atrophy, and marked hypomyelination, and a 3-month-old girl presenting with microcephaly, neonatal seizures, and marked psychomotor retardation. In both patients in vivo proton magnetic resonance spectroscopy (MRS) showed the presence of S-Ado signal at 8.3 ppm, consistent with a prior report. Interestingly, SAICAr signal was also detectable at 7.5 ppm in affected white matter, which has not been reported in vivo before. A novel splice-site mutation, c.IVS12 + 1/G > C, in the ADSL gene was identified in the second patient. Our findings confirm the utility of in vivo proton MRS in suggesting a specific diagnosis of ADSL deficiency, and also demonstrate an additional in vivo resonance (7.5 ppm) of SAICAr in the cases of severe disease. Copyright © 2012 Wiley Periodicals, Inc.

  19. Biophysical Studies of Function and Stability in Adenylosuccinate Lyase

    Science.gov (United States)

    Ray, Stephen; Duval, Nathan; Wilkinson, Terry, II; Shaheen, Sean; Ghosh, Kinshuk; Patterson, David

    2012-10-01

    Adenylosuccinate Lyase (ADSL) is a homotetrameric protein with four active sites that accommodate two reactions in the de novo purine biosynthesis pathway. It catalyzes the conversion of SAICAR to AICAR and AMPS to AMP. Point mutations in the gene encoding the protein ADSL lead to ADSL deficiency, a disorder characterized by serious neurological and physiological symptoms. Two leading hypotheses regarding the pathogenesis are ``Loss of Function'' or ``Gain in Toxic Function.'' These hypotheses can be related to the reduction of either the enzyme kinetics or the stability of the tetramer structure. Enzymatic studies can be used to provide a quantitative measure of the extent to which the enzyme acts on its designated substrates, SAICAR and AMPS. Recent kinetic studies have measured activity only on the substrates independently. Here we present characterization of enzyme kinetics for the biophysically interesting and physiologically relevant case where two substrates exhibit competitive binding to the enzyme, for both wild type and disease causing mutants of ADSL. Preliminary data suggest equivalent specific activities may be necessary to suppress severe phenotypes from expressing. Additionally, we will present results on the role of mutations on the thermodynamic stability of the enzyme. We will discuss thermodynamic analysis that gives a direct quantitative measure of the propensity of formation of folded and unfolded states of the protein, which influence the functionality of the protein and may also influence aggregation.

  20. Cystathionine γ-lyase deficiency mediates neurodegeneration in Huntington's disease.

    Science.gov (United States)

    Paul, Bindu D; Sbodio, Juan I; Xu, Risheng; Vandiver, M Scott; Cha, Jiyoung Y; Snowman, Adele M; Snyder, Solomon H

    2014-05-01

    Huntington's disease is an autosomal dominant disease associated with a mutation in the gene encoding huntingtin (Htt) leading to expanded polyglutamine repeats of mutant Htt (mHtt) that elicit oxidative stress, neurotoxicity, and motor and behavioural changes. Huntington's disease is characterized by highly selective and profound damage to the corpus striatum, which regulates motor function. Striatal selectivity of Huntington's disease may reflect the striatally selective small G protein Rhes binding to mHtt and enhancing its neurotoxicity. Specific molecular mechanisms by which mHtt elicits neurodegeneration have been hard to determine. Here we show a major depletion of cystathionine γ-lyase (CSE), the biosynthetic enzyme for cysteine, in Huntington's disease tissues, which may mediate Huntington's disease pathophysiology. The defect occurs at the transcriptional level and seems to reflect influences of mHtt on specificity protein 1, a transcriptional activator for CSE. Consistent with the notion of loss of CSE as a pathogenic mechanism, supplementation with cysteine reverses abnormalities in cultures of Huntington's disease tissues and in intact mouse models of Huntington's disease, suggesting therapeutic potential.

  1. Sexual Dimorphism in the Selenocysteine Lyase Knockout Mouse

    Directory of Open Access Journals (Sweden)

    Ashley N. Ogawa-Wong

    2018-01-01

    Full Text Available Selenium (Se is an essential micronutrient known for its antioxidant properties and health benefits, attributed to its presence in selenoproteins as the amino acid, selenocysteine. Selenocysteine lyase (Scly catalyzes hydrolysis of selenocysteine to selenide and alanine, facilitating re-utilization of Se for de novo selenoprotein synthesis. Previously, it was reported that male Scly−/− mice develop increased body weight and body fat composition, and altered lipid and carbohydrate metabolism, compared to wild type mice. Strikingly, females appeared to present with a less severe phenotype, suggesting the relationship between Scly and energy metabolism may be regulated in a sex-specific manner. Here, we report that while body weight and body fat gain occur in both male and female Scly−/− mice, strikingly, males are susceptible to developing glucose intolerance, whereas female Scly−/− mice are protected. Because Se is critical for male reproduction, we hypothesized that castration would attenuate the metabolic dysfunction observed in male Scly−/− mice by eliminating sequestration of Se in testes. We report that fasting serum insulin levels were significantly reduced in castrated males compared to controls, but islet area was unchanged between groups. Finally, both male and female Scly−/− mice exhibit reduced hypothalamic expression of selenoproteins S, M, and glutathione peroxidase 1.

  2. Phenol recovery with liquid ammonia

    Energy Technology Data Exchange (ETDEWEB)

    Heilman

    1943-08-03

    This report covers the results of a phenol recovery plant at Ludwigshafen which had recently suffered a severe explosion. From a Gelsenberg hydrogenation middle oil with 18 percent phenol, an 86 to 96 percent phenol fraction was recovered. Because the occurring neutral oil was phenol-free and because with the process a phenol loss was impossible, it was assumed that the yield was quantitative. With regard to the working process, the middle oil was fed into the upper section of a column four feet high, in which liquid ammonia from below climbed upward. The ammonia thereby absorbed the phenol quantitatively, and the ammonium phenolate solution absorbed a certain amount of neutral oil. The loaded ammonia went over the top of the column while at the foot of the column the phenol-free neutral oil collected and was drawn off. The ammonium phenolate solution was then washed with light gasoline in a second column. For this, the ammonia was fed into the upper, the light gasoline into the lower part of the column. The light gasoline absorbed almost quantitatively the neutral oil which was molecularly or actually colloidally dissolved in the ammonium phenolate solution, and even a small amount of the phenol and ammonia. Thickening in concentration, the light gasoline was fed into a storage tank where it was freed of its dissolved components by atmospheric distillation and recycled into the process. The ammonium phenolate solution which before the gasoline wash left behind a vaporization residue with about 40 percent phenol afterwards produced a raw phenol of 86 to 96 percent pure phenol. Because of technical difficulties, the concentration of the washed ammonium phenolate solution could not be determined. It was gathered at the bottom of the second column and fed into a storage tank where the phenol was freed by pressure distillation. The ammonia was then recycled into the process.

  3. Ammonia abatement by slurry acidification

    DEFF Research Database (Denmark)

    Petersen, Søren O.; Hutchings, Nicholas J.; Hafner, Sasha D.

    2016-01-01

    Livestock production systems can be major sources of trace gases including ammonia (NH3), the greenhouse gases methane (CH4) and nitrous oxide (N2O), and odorous compounds such as hydrogen sulphide (H2S). Short-term campaigns have indicated that acidification of livestock slurry during in...... sections with 30-32 pigs with or without daily adjustment of slurry pH to below 6. Ammonia losses from reference sections with untreated slurry were between 9.5 and 12.4% of N excreted, and from sections with acidified slurry between 3.1 and 6.2%. Acidification reduced total emissions of NH3 by 66 and 71...

  4. Catalytic Organometallic Reactions of Ammonia

    Science.gov (United States)

    Klinkenberg, Jessica L.

    2012-01-01

    Until recently, ammonia had rarely succumbed to catalytic transformations with homogeneous catalysts, and the development of such reactions that are selective for the formation of single products under mild conditions has encountered numerous challenges. However, recently developed catalysts have allowed several classes of reactions to create products with nitrogen-containing functional groups from ammonia. These reactions include hydroaminomethylation, reductive amination, alkylation, allylic substitution, hydroamination, and cross-coupling. This Minireview describes examples of these processes and the factors that control catalyst activity and selectivity. PMID:20857466

  5. Ammonia synthesis at low temperatures

    DEFF Research Database (Denmark)

    Rod, Thomas Holm; Logadottir, Ashildur; Nørskov, Jens Kehlet

    2000-01-01

    Density functional theory (DFT) calculations of reaction paths and energies for the industrial and the biological catalytic ammonia synthesis processes are compared. The industrial catalyst is modeled by a ruthenium surface, while the active part of the enzyme is modeled by a MoFe6S9 complex...... have been carried out to evaluate its feasibility. The calculations suggest that it might be possible to catalytically produce ammonia from molecular nitrogen at low temperatures and pressures, in particular if energy is fed into the process electrochemically. (C) 2000 American Institute of Physics....

  6. Ammonia-water Rankine cycle

    International Nuclear Information System (INIS)

    Bo Hanliang; Ma Changwen; Wu Shaorong

    1997-01-01

    On characteristics of heating source and cooling source in nuclear heating reactor cooperation, the authors advance a new kind of power cycle in which a multicomponent mixture as the work fluid, ammonia-water Rankine cycle, describe its running principle, and compare it with steam Rankine cycle in the same situation. The result is that: the new kind of power cycle, ammonia-water Rankine cycle has higher electricity efficiency; it suits for the situation of heating source and cooling source which offered by nuclear heating reactor cooperation. For low temperature heating source, it maybe has a widely application

  7. RESULTS OF INITIAL AMMONIA OXIDATION TESTING

    Energy Technology Data Exchange (ETDEWEB)

    Nash, C. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL); Fowley, M. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)

    2016-12-30

    This memo presents an experimental survey of aqueous phase chemical processes to remove aqueous ammonia from waste process streams. Ammonia is generated in both the current Hanford waste flowsheet and in future waste processing. Much ammonia will be generated in the Low Activity Waste (LAW) melters.i Testing with simulants in glass melters at Catholic University has demonstrated the significant ammonia production.ii The primary reaction there is the reducing action of sugar on nitrate in the melter cold cap. Ammonia has been found to be a problem in secondary waste stabilization. Ammonia vapors are noxious and destruction of ammonia could reduce hazards to waste treatment process personnel. It is easily evolved especially when ammonia-bearing solutions are adjusted to high pH.

  8. The Chinese soft-shelled turtle, Pelodiscus sinensis, decreases nitrogenous excretion, reduces urea synthesis and suppresses ammonia production during emersion.

    Science.gov (United States)

    Ip, Yuen K; Lee, Serene M L; Wong, Wai P; Chew, Shit F

    2013-05-01

    The objective of this study was to examine the effects of 6 days of emersion on nitrogen metabolism and excretion in the Chinese soft-shelled turtle, Pelodiscus sinensis. Despite having a soft shell with a cutaneous surface that is known to be water permeable, P. sinensis lost only ~2% of body mass and was able to maintain its hematocrit and plasma osmolality, [Na(+)] and [Cl(-)] during 6 days of emersion. During emersion, it ameliorated water loss by reducing urine output, which led to a reduction (by 29-76%) in ammonia excretion. In comparison, there was a more prominent reduction (by 82-99%) in urea excretion during emersion due to a lack of water to flush the buccopharyngeal epithelium, which is known to be the major route of urea excretion. Consequently, emersion resulted in an apparent shift from ureotely to ammonotely in P. sinensis. Although urea concentration increased in several tissues, the excess urea accumulated could only account for 13-22% of the deficit in urea excretion. Hence, it can be concluded that a decrease (~80%) in urea synthesis occurred in P. sinensis during the 6 days of emersion. Indeed, emersion led to significant decreases in the activity of some ornithine-urea cycle enzymes (argininosuccinate synthetase/argininosuccinate lyase and arginase) from the liver of P. sinensis. As a decrease in urea synthesis occurred without the accumulation of ammonia and total free amino acids, it can be deduced that ammonia production through amino acid catabolism was suppressed with a proportional reduction in proteolysis in P. sinensis during emersion. Indeed, calculated results revealed that there could be a prominent decrease (~88%) in ammonia production in turtles after 6 days of emersion. In summary, despite being ureogenic and ureotelic in water, P. sinensis adopted a reduction in ammonia production, instead of increased urea synthesis, as the major strategy to ameliorate ammonia toxicity and problems associated with dehydration during

  9. Clinical utility of breath ammonia for evaluation of ammonia physiology in healthy and cirrhotic adults

    OpenAIRE

    Spacek, Lisa A; Mudalel, Matthew; Tittel, Frank; Risby, Terence H; Solga, Steven F

    2015-01-01

    Blood ammonia is routinely used in clinical settings to assess systemic ammonia in hepatic encephalopathy and urea cycle disorders. Despite its drawbacks, blood measurement is often used as a comparator in breath studies because it is a standard clinical test. We sought to evaluate sources of measurement error and potential clinical utility of breath ammonia compared to blood ammonia.

  10. Determination of ammonia in ear-lobe capillary blood is an alternative to arterial blood ammonia

    NARCIS (Netherlands)

    Huizenga, J. R.; Gips, C. H.; Conn, H. O.; Jansen, P. L.

    1995-01-01

    Blood ammonia determination is a laboratory test to diagnose hepatic encephalopathy. Arterial blood is superior to peripheral venous blood ammonia because of ammonia metabolism in muscle. We have compared capillary with arterial whole blood ammonia as capillary sampling is an attractive alternative.

  11. DETERMINATION OF AMMONIA IN EAR-LOBE CAPILLARY BLOOD IS AN ALTERNATIVE TO ARTERIAL BLOOD AMMONIA

    NARCIS (Netherlands)

    HUIZENGA, [No Value; GIPS, CH; CONN, HO; JANSEN, PLM

    1995-01-01

    Blood ammonia determination is a laboratory test to diagnose hepatic encephalopathy. Arterial blood is superior to peripheral venous blood ammonia because of ammonia metabolism in muscle. We have compared capillary with arterial whole blood ammonia as capillary sampling is an attractive alternative.

  12. Solvent Isotope-induced Equilibrium Perturbation for Isocitrate Lyase

    Science.gov (United States)

    Quartararo, Christine E.; Hadi, Timin; Cahill, Sean M.; Blanchard, John S.

    2014-01-01

    Isocitrate lyase (ICL) catalyzes the reversible retro-aldol cleavage of isocitrate to generate glyoxylate and succinate. ICL is the first enzyme of the glyoxylate shunt, which allows for the anaplerosis of citric acid cycle intermediates under nutrient limiting conditions. In Mycobacterium tuberculosis, the source of ICL for these studies, ICL is vital for the persistence phase of the bacteria’s life cycle. Solvent kinetic isotope effects (KIEs) in the direction of isocitrate cleavage of D2OV = 2.0 ± 0.1 and D2O[V/Kisocitrate] = 2.2 ± 0.3 arise from the initial deprotonation of the C2 hydroxyl group of isocitrate or the protonation of the aci-acid of succinate product of the isocitrate aldol cleavage by a solvent-derived proton. This KIE suggested that an equilibrium mixture of all protiated isocitrate, glyoxylate and succinate prepared in D2O, would undergo transient changes in equilibrium concentrations as a result of the solvent KIE and solvent-derived deuterium incorporation into both succinate and isocitrate. No change in the isotopic composition of glyoxylate was expected or observed. We have directly monitored the changing concentrations of all isotopic species of all reactants and products using a combination of NMR spectroscopy and mass spectrometry. Continuous monitoring of glyoxylate by 1H NMR spectroscopy shows a clear equilibrium perturbation in D2O. The final equilibrium isotopic composition of reactants in D2O revealed di-deuterated succinate, protiated glyoxylate, and mono-deuterated isocitrate, with the transient appearance and disappearance of mono-deuterated succinate. A model for the equilibrium perturbation of substrate species, and their time-dependent isotopic composition is presented. PMID:24261638

  13. Novel features in the evolution of adenylosuccinate lyase deficiency.

    Science.gov (United States)

    Pérez-Dueñas, Belén; Sempere, Angela; Campistol, Jaume; Alonso-Colmenero, Itziar; Díez, María; González, Verónica; Merinero, Begoña; Desviat, Lourdes R; Artuch, Rafael

    2012-07-01

    Adenylosuccinate lyase (ADSL) deficiency is an autosomal recessive disorder of the purine synthesis which results in accumulation of succinylpurines (succinyladenosine (S-Ado) and succinylamino-imidazole carboxamide riboside (SAICAr)) in body fluids. Patients present developmental delay, often accompanied by epilepsy and autistic spectrum disorders. To describe atypical neurological features in the evolution of three novel unrelated cases of ADSL deficiency. A 9-year-old boy with severe cognitive impairment and autistic behaviour received d-ribose therapy for one year. Drug withdrawal was associated with acute neurological deterioration, severe brain atrophy and demyelination on MRI. The second patient is a 5.5-year-old girl with mild developmental delay who presented a benign course with moderate cognitive impairment as the only feature in her evolution. The final patient is a 14-year-old boy with severe cognitive impairment who developed drug-resistant epilepsy and bathing reflex seizures, progressive spasticity in the lower limbs and thoracic deformity. SAICAr and S-Ado in urine were analysed by HPLC with diode array detection. Diagnosis was confirmed by molecular analysis of the ADSL gene. An elevation of S-Ado and SAICAr excretion in urine was detected in all three patients. The patients were homozygous for the missence change p.I369L and for the novel change p.M389V. Drug-resistant epilepsy and specific therapeutic interventions may modify the neurological outcome in ADSL deficiency. d-ribose must be considered with caution as, in our experience, it returns no clinical benefit and drug withdrawal can precipitate status epilepticus and acute neurological deterioration. Copyright © 2011 European Paediatric Neurology Society. Published by Elsevier Ltd. All rights reserved.

  14. Clinical, biochemical and molecular genetic correlations in adenylosuccinate lyase deficiency.

    Science.gov (United States)

    Race, V; Marie, S; Vincent, M F; Van den Berghe, G

    2000-09-01

    Adenylosuccinate lyase (ADSL) deficiency (MIM 103050) is an autosomal recessive inborn error of purine synthesis characterized by the accumulation in body fluids of succinylaminoimidazolecarboxamide (SAICA) riboside and succinyladenosine (S-Ado), the dephosphorylated derivatives of the two substrates of the enzyme. Because ADSL-deficient patients display widely variable degrees of psychomotor retardation, we have expressed eight mutated ADSL enzymes as thioredoxin fusions and compared their properties with the clinical and biochemical characteristics of 10 patients. Three expressed mutated ADSL enzymes (M26L, R426H and T450S) were thermolabile, four (A2V, R141W, R303C and S395R) were thermostable and one (del206-218), was inactive. Thermolabile mutations decreased activities with SAICA ribotide (SAICAR) and adenylosuccinate (S-AMP) in parallel, or more with SAICAR than with S-AMP. Patients homozygous for one of these mutations, R426H, displayed similarly decreased ADSL activities in their fibroblasts, S-Ado:SAICA riboside ratios of approximately 1 in their cerebrospinal fluid and were profoundly retarded. With the exception of A2V, thermostable mutations decreased activity with S-AMP to a much more marked extent than with SAICAR. Two unrelated patients homozygous for one of the thermostable mutations, R303C, also displayed a much more marked decrease in the activity of fibroblast ADSL with S-AMP than with SAICAR, had S-Ado:SAICA riboside ratios between 3 and 4 in their cerebrospinal fluid and were mildly retarded. These results suggest that, in some cases, the genetic lesion of ADSL determines the ratio of its activities with S-AMP versus SAICAR, which in turn defines the S-Ado:SAICA riboside ratio and the patients' mental status.

  15. 46 CFR 154.1760 - Liquid ammonia.

    Science.gov (United States)

    2010-10-01

    ... 46 Shipping 5 2010-10-01 2010-10-01 false Liquid ammonia. 154.1760 Section 154.1760 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES SAFETY STANDARDS FOR....1760 Liquid ammonia. The master shall ensure that no person sprays liquid ammonia into a cargo tank...

  16. 27 CFR 21.96 - Ammonia, aqueous.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Ammonia, aqueous. 21.96 Section 21.96 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT... Ammonia, aqueous. (a) Alkalinity. Strongly alkaline to litmus. (b) Ammonia content. 27 to 30 percent by...

  17. Synthesis of ammonia with microwave plasma

    International Nuclear Information System (INIS)

    Xu Wenguo; Yu Aimin; Liu Jun; Jin Qinhan

    1991-01-01

    THe synthesis of ammonia absorbed on 13X zeolite with the aid of microwave plasma is described. The ammonia molecule absorbed on 13X zeolite as ammonium ions were detected by IR spectroscopy. The results obtained show that the ammonia synthesis is facilitated by the surface reactions of NH x (x = 1, 2) radicals adsorbed on zeolite with hydrogen atoms

  18. Hydrogen production using ammonia borane

    Science.gov (United States)

    Hamilton, Charles W; Baker, R. Thomas; Semelsberger, Troy A; Shrestha, Roshan P

    2013-12-24

    Hydrogen ("H.sub.2") is produced when ammonia borane reacts with a catalyst complex of the formula L.sub.nM-X wherein M is a base metal such as iron, X is an anionic nitrogen- or phosphorus-based ligand or hydride, and L is a neutral ancillary ligand that is a neutral monodentate or polydentate ligand.

  19. Reducing ammonia emissions in Europe

    DEFF Research Database (Denmark)

    Jacobsen, Brian H.

    2011-01-01

    The NEC directive has set targets for the 2010 ammonia emissions from a number of European countries. The target will be reached by most EU-countries and the total emission for EU-27 has been reduced by 22% from 1990 to 2007. Denmark is one of the countries with the largest reductions since 1990...

  20. by thiocyanates in liquid ammonia

    African Journals Online (AJOL)

    ABSTRACT. Infrared and Raman spectroscopy techniques have been used to study the ionic interactions of strontium(II) and barium(II) with thiocyanate ion in liquid ammonia. A number of bands were observed in both ν(CN) and ν(CS) regions of infrared and Raman spectra and these were assigned to 1:1 contact ion pair, ...

  1. Haber Process for Ammonia Synthesis

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 16; Issue 12. Haber Process for Ammonia Synthesis. Jayant M Modak. Volume 16 Issue 12 December 2011 pp 1159-1167. Fulltext. Click here to view fulltext PDF. Permanent link: https://www.ias.ac.in/article/fulltext/reso/016/12/1159-1167. Keywords.

  2. Haber Process for Ammonia Synthesis

    Indian Academy of Sciences (India)

    Before synthetic nitrogen fixation, wastes and manures of various types or their decomposition products, and ammonium sulfate, which is a by-product from the coking of coal, were the primary sources of agricultural nitrogen. Chilean saltpetre, saltpetre from hu- man and animal urine, and later ammonia recovered from coke.

  3. Probing the structure of glucan lyases – the lytic members of GH31 - by sequence analysis, circular dichroism and proteolysis

    DEFF Research Database (Denmark)

    Ernst, Heidi; Lo Leggio, Leila; Yu, Shukun

    2005-01-01

    Glucan lyase (GL) is a polysaccharide lyase with unique characteristics. It is involved in an alternative pathway for the degradation of alpha-glucans, the anhydrofructose pathway. Sequence similarity suggests that this lytic enzyme belongs to glycoside hydrolase family 31, for which until very r...

  4. Transcriptional Response of the Archaeal Ammonia Oxidizer Nitrosopumilus maritimus to Low and Environmentally Relevant Ammonia Concentrations

    OpenAIRE

    Nakagawa, Tatsunori; Stahl, David A.

    2013-01-01

    The ability of chemoautotrophic ammonia-oxidizing archaea to compete for ammonia among marine microorganisms at low ambient concentrations has been in part attributed to their extremely high affinity for ammonia, but as yet there is no mechanistic understanding of supporting metabolism. We examined transcription of selected genes for anabolic functions (CO2 fixation, ammonia transport, and cell wall synthesis) and a central catabolic function (ammonia oxidation) in the thaumarchaeon Nitrosopu...

  5. Transcriptional Response of the Archaeal Ammonia Oxidizer Nitrosopumilus maritimus to Low and Environmentally Relevant Ammonia Concentrations

    Science.gov (United States)

    Stahl, David A.

    2013-01-01

    The ability of chemoautotrophic ammonia-oxidizing archaea to compete for ammonia among marine microorganisms at low ambient concentrations has been in part attributed to their extremely high affinity for ammonia, but as yet there is no mechanistic understanding of supporting metabolism. We examined transcription of selected genes for anabolic functions (CO2 fixation, ammonia transport, and cell wall synthesis) and a central catabolic function (ammonia oxidation) in the thaumarchaeon Nitrosopumilus maritimus SCM1 growing at two ammonia concentrations, as measured by combined ammonia and ammonium, one well above the Km for ammonia oxidation (∼500 μM) and the other well below the Km (ammonia-replete to ammonia-limiting conditions. Transcript levels for ammonia oxidation, CO2 fixation, and one of the ammonia transport genes were approximately the same at high and low ammonia availability. Transcripts for all analyzed genes decreased with time in the complete absence of ammonia, but with various rates of decay. The new steady-state mRNA levels established are presumably more reflective of the natural physiological state of ammonia-oxidizing archaea and offer a reference for interpreting message abundance patterns in the natural environment. PMID:23995944

  6. Adenylosuccinate lyase deficiency in the United Kingdom pediatric population: first three cases.

    Science.gov (United States)

    Lundy, Claire T; Jungbluth, Heinz; Pohl, Keith R E; Siddiqui, Ata; Marinaki, Anthony M; Mundy, Helen; Champion, Michael P

    2010-11-01

    Adenylosuccinate lyase deficiency is an autosomal recessive disorder of purine metabolism resulting from mutations in the ADSL gene on chromosome subband 22q13.1 and associated with a wide range of clinical manifestations. Although there is currently no effective treatment of ADSL deficiency, recognition of the condition is important, because prenatal genetic diagnosis can be offered to affected families. Reported here are the cases of the only three children diagnosed to date in the United Kingdom with adenylosuccinate lyase deficiency, to further delineate the clinical phenotype and to raise awareness of this disorder. Copyright © 2010 Elsevier Inc. All rights reserved.

  7. Ammonia toxicity: from head to toe?

    Science.gov (United States)

    Dasarathy, Srinivasan; Mookerjee, Rajeshwar P; Rackayova, Veronika; Rangroo Thrane, Vinita; Vairappan, Balasubramaniyan; Ott, Peter; Rose, Christopher F

    2017-04-01

    Ammonia is diffused and transported across all plasma membranes. This entails that hyperammonemia leads to an increase in ammonia in all organs and tissues. It is known that the toxic ramifications of ammonia primarily touch the brain and cause neurological impairment. However, the deleterious effects of ammonia are not specific to the brain, as the direct effect of increased ammonia (change in pH, membrane potential, metabolism) can occur in any type of cell. Therefore, in the setting of chronic liver disease where multi-organ dysfunction is common, the role of ammonia, only as neurotoxin, is challenged. This review provides insights and evidence that increased ammonia can disturb many organ and cell types and hence lead to dysfunction.

  8. Ammonia for hydrogen storage: challenges and opportunities

    DEFF Research Database (Denmark)

    Klerke, Asbjørn; Christensen, Claus H.; Nørskov, Jens Kehlet

    2008-01-01

    The possibility of using ammonia as a hydrogen carrier is discussed. Compared to other hydrogen storage materials, ammonia has the advantages of a high hydrogen density, a well-developed technology for synthesis and distribution, and easy catalytic decomposition. Compared to hydrocarbons...... and alcohols, it has the advantage that there is no CO2 emission at the end user. The drawbacks are mainly the toxicity of liquid ammonia and the problems related to trace amounts of ammonia in the hydrogen after decomposition. Storage of ammonia in metal ammine salts is discussed, and it is shown...... that this maintains the high volumetric hydrogen density while alleviating the problems of handling the ammonia. Some of the remaining challenges for research in ammonia as a hydrogen carrier are outlined....

  9. Operation experience with elevated ammonia

    International Nuclear Information System (INIS)

    Vankova, Katerina; Kysela, Jan; Malac, Miroslav; Petrecky, Igor; Svarc, Vladimir

    2011-01-01

    The 10 VVER units in the Czech and Slovak Republics are all in very good water chemistry and radiation condition, yet questions have arisen regarding the optimization of cycle chemistry and improved operation in these units. To address these issues, a comprehensive experimental program for different water chemistries of the primary circuit was carried out at the Rez Nuclear Research Institute, Czech Republic, with the goal of judging the influence of various water chemistries on radiation build-up. Four types of water chemistries were compared: standard VVER water chemistry (in common use), direct hydrogen dosing without ammonia, standard VVER water chemistry with elevated ammonia levels, and zinc dosing to standard VVER water chemistry. The test results showed that the types of water chemistry other than the common one have benefits for the operation of the nuclear power plant (NPP) primary circuit. Operation experience with elevated ammonia at NPP Dukovany Units 3 and 4 is presented which validates the experimental results, demonstrating improved corrosion product volume activity. (orig.)

  10. Planar waveguide sensor of ammonia

    Science.gov (United States)

    Rogoziński, Roman; Tyszkiewicz, Cuma; Karasiński, Paweł; Izydorczyk, Weronika

    2015-12-01

    The paper presents the concept of forming ammonia sensor based on a planar waveguide structure. It is an amplitude sensor produced on the basis of the multimode waveguide. The technological base for this kind of structure is the ion exchange method and the sol-gel method. The planar multimode waveguide of channel type is produced in glass substrate (soda-lime glass of Menzel-Glaser company) by the selective Ag+↔Na+ ion exchange. On the surface of the glass substrate a porous (~40%) silica layer is produced by the sol-gel method. This layer is sensitized to the presence of ammonia in the surrounding atmosphere by impregnation with Bromocresol Purple (BCP) dye. Therefore it constitutes a sensor layer. Spectrophotometric tests carried out showed about 50% reduction of cross-transmission changes of such sensor layer for a wave λ=593 nm caused by the presence of 25% ammonia water vapor in its ambience. The radiation source used in this type of sensor structure is a light emitting diode LED. The gradient channel waveguide is designed for frontal connection (optical glue) with a standard multimode telecommunications waveguide 62.5/125μm.

  11. Exergy analysis of industrial ammonia synthesis

    International Nuclear Information System (INIS)

    Kirova-Yordanova, Zornitza

    2004-01-01

    Exergy consumption of ammonia production plants depends strongly on the ammonia synthesis loop design. Due to the thermodynamically limited low degree of conversion of hydrogen-nitrogen mixture to ammonia, industrial ammonia synthesis is implemented as recycle process (so-called 'ammonia synthesis loop'). Significant quantities of reactants are recycled back to reactor, after the removal of ammonia at low temperatures. Modern ammonia synthesis plants use well-developed heat- and cold recovery to improve the reaction heat utilisation and to reduce the refrigeration costs. In this work, the exergy method is applied to estimate the effect of the most important process parameters on the exergy efficiency of industrial ammonia synthesis. A specific approach, including suitable definitions of the system boundaries and process parameters, is proposed. Exergy efficiency indexes are discussed in order to make the results applicable to ammonia synthesis loops of various designs. The dependence of the exergy losses on properly selected independent process parameters is studied. Some results from detailed exergy analysis of the most commonly used ammonia synthesis loop design configurations at a wide range of selected parameters values are shown

  12. Purification and characterization of a new bioscouring pectate lyase from Bacillus pumilus BK2

    NARCIS (Netherlands)

    Klug-Santner, Barbara G.; Schnitzhofer, Wolfgang; Vrsanska, Maria; Weber, Jörg; Agrawal, Pramod; Nierstrasz, Vincent; Guebitz, Georg M.

    2006-01-01

    An alkalophilic bacterium was isolated based on the potential of extra-cellular enzymes for bioscouring. The bacterium was identified as a new strain of Bacillus pumilus BK2 producing an extra-cellular endo-pectate lyase PL (EC 4.2.2.2). PL was purified to homogeneity in three steps and has a

  13. Biotechnological potential of agro-industrial waste in the synthesis of pectin lyase from Aspergillus brasiliensis.

    Science.gov (United States)

    Pili, Jonaina; Danielli, Angélica; Nyari, Nádia Ld; Zeni, Jamile; Cansian, Rogério L; Backes, Geciane T; Valduga, Eunice

    2018-03-01

    This study aims at investigating pectin lyase bioproduction in submerged fermentation with synthetic medium and agro-industrial residues, using the filamentous fungus Aspergillus brasiliensis. The maximum pectin lyase activity in a synthetic medium (42 g/l pectin, 40 g/l yeast extract, and 0.02 g/l iron sulfate) was 31 U/ml, and 46 U/ml in the agro-industrial medium (160 g/l orange peel, 150 g/l corn steep liquor, and 300 g/l parboiled rice water), obtained over 60 and 124 h of bioproduction, 180 r/min, 30 ℃, pH initial 5.5, and 5·10 6 spores/ml, respectively. Partial characterization of pectin lyase crude enzyme extract obtained from the synthetic medium and the one made of agro-industrial residues showed optimum conditions at pH of 5.5 and 4.5 and temperatures of 37 and 55 ℃, respectively. The E d obtained was 3.13 and 9.15 kJ/mol, and the half-life time (t 1/2 ) was 5.71 and 80 h at 55 ℃ for pectin lyase produced in synthetic and agro-industrial medium, respectively.

  14. Cystathionine-γ-lyase gene silencing with siRNA in monocytes ...

    Indian Academy of Sciences (India)

    Hydrogen sulphide is an endogenous inflammatory mediator produced by cystathionine-γ-lyase (CSE) in macrophages. To determine the role of H2S and macrophages in sepsis, we used small interference RNA (siRNA) to target the CSE gene and investigated its effect in a mouse model of sepsis. Cecal ligation puncture ...

  15. Hematopoietic sphingosine 1-phosphate lyase deficiency decreases atherosclerotic lesion development in LDL-receptor deficient mice.

    Directory of Open Access Journals (Sweden)

    Martine Bot

    Full Text Available AIMS: Altered sphingosine 1-phosphate (S1P homeostasis and signaling is implicated in various inflammatory diseases including atherosclerosis. As S1P levels are tightly controlled by S1P lyase, we investigated the impact of hematopoietic S1P lyase (Sgpl1(-/- deficiency on leukocyte subsets relevant to atherosclerosis. METHODS AND RESULTS: LDL receptor deficient mice that were transplanted with Sgpl1(-/- bone marrow showed disrupted S1P gradients translating into lymphopenia and abrogated lymphocyte mitogenic and cytokine response as compared to controls. Remarkably however, Sgpl1(-/- chimeras displayed mild monocytosis, due to impeded stromal retention and myelopoiesis, and plasma cytokine and macrophage expression patterns, that were largely compatible with classical macrophage activation. Collectively these two phenotypic features of Sgpl1 deficiency culminated in diminished atherogenic response. CONCLUSIONS: Here we not only firmly establish the critical role of hematopoietic S1P lyase in controlling S1P levels and T cell trafficking in blood and lymphoid tissue, but also identify leukocyte Sgpl1 as critical factor in monocyte macrophage differentiation and function. Its, partly counterbalancing, pro- and anti-inflammatory activity spectrum imply that intervention in S1P lyase function in inflammatory disorders such as atherosclerosis should be considered with caution.

  16. Biochemical and structural characterization of a novel bacterial manganese-dependent hydroxynitrile lyase.

    NARCIS (Netherlands)

    Hajnal, I.; Lyskowski, A.; Hanefeld, U.; Gruber, K.; Schwab, H.; Steiner, K.

    2013-01-01

    Hydroxynitrile lyases (HNLs), which catalyse the decomposition of cyanohydrins, are found mainly in plants. In vitro, they are able to catalyse the synthesis of enantiopure cyanohydrins, which are versatile building blocks in the chemical industry. Recently, HNLs have also been discovered in

  17. Novel Pectate Lyase Genes of Heterodera glycines Play Key Roles in the Early Stage of Parasitism.

    Directory of Open Access Journals (Sweden)

    Huan Peng

    Full Text Available Pectate lyases are known to play a key role in pectin degradation by catalyzing the random cleavage of internal polymer linkages (endo-pectinases. In this paper, four novel cDNAs, designated Hg-pel-3, Hg-pel-4, Hg-pel-6 and Hg-pel-7, that encode pectate lyases were cloned and characterized from the soybean cyst nematode, Heterodera glycines. The predicted protein sequences of HG-PEL-3, HG-PEL-4 and HG-PEL-6 differed significantly in both their amino acid sequences and their genomic structures from other pectate lyases of H. glycines (HG-PEL-1, HG-PEL-2 and HG-PEL-7. A phylogenetic study revealed that the pectate lyase proteins of H. glycines are clustered into distinct clades and have distinct numbers and positioning of introns, which suggests that the pectate lyase genes of H. glycines may have evolved from at least two ancestral genes. A Southern blot analysis revealed that multiple Hg-pel-6-like genes were present in the H. glycines genome. In situ hybridization showed that four novel pectate lyases (Hg-pel-3, Hg-pel-4, Hg-pel-6 and Hg-pel-7 were actively transcribed in the subventral esophageal gland cells. A semi-quantitative RT-PCR assay supported the finding that the expression of these genes was strong in the egg, pre-parasitic second-stage juvenile (J2 and early parasitic J2 stages and that it declined in further developmental stages of the nematode. This expression pattern suggests that these proteins play a role in the migratory phase of the nematode life cycle. Knocking down Hg-pel-6 using in vitro RNA interference resulted in a 46.9% reduction of the number of nematodes that invaded the plants and a 61.5% suppression of the development of H. glycines females within roots compared to the GFP-dsRNA control. Plant host-derived RNAi induced the silencing of the Hg-pel-6gene, which significantly reduced the nematode infection levels at 7 Days post inoculation (dpi. Similarly, this procedure reduced the number of female adults at 40 dpi

  18. Contribution of ammonia-oxidizing archaea and ammonia-oxidizing bacteria to ammonia oxidation in two nitrifying reactors.

    Science.gov (United States)

    Srithep, Papitchaya; Pornkulwat, Preeyaporn; Limpiyakorn, Tawan

    2018-03-01

    In this study, two laboratory nitrifying reactors (NRI and NRII), which were seeded by sludge from different sources and operated under different operating conditions, were found to possess distinct dominant ammonia-oxidizing microorganisms. Ammonia-oxidizing archaeal (AOA) amoA genes outnumbered ammonia-oxidizing bacterial (AOB) amoA genes in reactor NRI, while only AOB amoA genes were detectable in reactor NRII. The AOA amoA gene sequences retrieved from NRI were characterized within the Nitrososphaera sister cluster of the group 1.1b Thaumarchaeota. Two inhibitors for ammonia oxidation, allylthiourea (ATU) and 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (PTIO), were applied individually and as a mixture to observe the ammonia-oxidizing activity of both microorganisms in the reactors' sludge. The results indicated that AOA and AOB jointly oxidized ammonia in NRI, while AOB played the main role in ammonia oxidation in NRII. DNA-stable isotope probing with labeled 13 C-HCO 3 - was performed on NRI sludge. Incorporation of 13 C into AOA and AOB implied that both microorganisms may perform autotrophy during ammonia oxidation. Taken together, the results from this study provide direct evidence demonstrating the contribution of AOA and AOB to ammonia oxidation in the nitrifying reactors.

  19. Ammonia Process by Pressure Swing Adsorption

    Energy Technology Data Exchange (ETDEWEB)

    Dr Felix Jegede

    2010-12-27

    The overall objective of the project is to design, develop and demonstrate a technically feasible and commercially viable system to produce ammonia along with recovery of the products by adsorption separation methods and significantly decrease the energy requirement in ammonia production. This is achieved through a significantly more efficient ammonia psa recovery system. The new ammonia recovery system receives the reactor effluents and achieves complete ammonia recovery, (which completely eliminates the energy intensive refrigeration and condensation system currently used in ammonia production). It also recovers the unused reactants and recycles them back to the reactor, free of potential reactor contaminants, and without the need for re-compression and re-heat of recycle stream thereby further saving more energy. The result is a significantly lower energy consumption, along with capital cost savings.

  20. Ammonia emission from crop residues : quantification of ammonia volatilization based on crop residue properties

    NARCIS (Netherlands)

    Ruijter, de F.J.; Huijsmans, J.F.M.

    2012-01-01

    This paper gives an overview of available literature data on ammonia volatilization from crop residues. From these data, a relation is derived for the ammonia emission depending on the N-content of crop residue.

  1. New findings on cerebral ammonia uptake in HE using functional (13)N-ammonia PET

    DEFF Research Database (Denmark)

    Sørensen, Michael; Keiding, Susanne

    2007-01-01

    ) analysed such data in patients with HE by a kinetic model accounting for transfer of (13)N-ammonia across the blood-brain barrier (BBB) and intracellular formation of (13)N-glutamine. Initial unidirectional (13)N-ammonia transfer across BBB was characterized by the permeability-surface area product PS...... metabolites was linearly correlated to arterial ammonia. In conclusion, basic brain ammonia kinetics was not changed significantly in patients with cirrhosis +/- HE compared to healthy controls. Blood ammonia seems to be the more important factor for increased brain ammonia uptake in HE. Udgivelsesdato: 2007......PET is a functional imaging technique suitable for studies of brain ammonia metabolism. Dynamic (13)N-ammonia PET yields time-courses of radioactivity concentrations in brain (PET camera) and blood (samples). Ahl et al. (Hepatology 40:73-79, 2004) and Keiding et al. (Hepatology 43:42-50, 2006...

  2. 21 CFR 862.1065 - Ammonia test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Ammonia test system. 862.1065 Section 862.1065....1065 Ammonia test system. (a) Identification. An ammonia test system is a device intended to measure ammonia levels in blood, serum, and plasma, Ammonia measurements are used in the diagnosis and treatment...

  3. Synthesis of ammonia using sodium melt

    OpenAIRE

    Kawamura, Fumio; Taniguchi, Takashi

    2017-01-01

    Research into inexpensive ammonia synthesis has increased recently because ammonia can be used as a hydrogen carrier or as a next generation fuel which does not emit CO2. Furthermore, improving the efficiency of ammonia synthesis is necessary, because current synthesis methods emit significant amounts of CO2. To achieve these goals, catalysts that can effectively reduce the synthesis temperature and pressure, relative to those required in the Haber-Bosch process, are required. Although severa...

  4. Polyaniline-based optical ammonia detector

    Science.gov (United States)

    Duan, Yixiang; Jin, Zhe; Su, Yongxuan

    2002-01-01

    Electronic absorption spectroscopy of a polyaniline film deposited on a polyethylene surface by chemical oxidation of aniline monomer at room temperature was used to quantitatively detect ammonia gas. The present optical ammonia gas detector was found to have a response time of less than 15 s, a regeneration time of less than 2 min. at room temperature, and a detection limit of 1 ppm (v/v) for ammonia, with a linear dynamic range from 180 ppm to 18,000 ppm.

  5. Ammonia separation in novel bio filters

    Directory of Open Access Journals (Sweden)

    Jochen Hahne

    2017-04-01

    Full Text Available Currently used bio filters are suited for odor reduction in livestock keeping but not for ammonia separation. Therefore it was the objective to develop a novel bio filter system which was able to ensure a high and long-lasting ammonia separation. This novel bio filter, which is equipped with a pH control in a water swamp beneath the filter layer and a conductivity control for water discharge, was investigated in terms of ammonia separation and nitrogen disposition over several months under practical conditions. The results show a stable ammonia separation of more than 88 % if certain consecutively described operating conditions are kept.

  6. Sizing of an Ammonia Discharge Tank

    Directory of Open Access Journals (Sweden)

    Tuliagenda Beckfords

    2011-01-01

    Full Text Available Phosphate companies use well-stirred tanks to regulate the concentration of ammonia they discharge via their wastewater, preventing ammonia spikes from exceeding the cap set by the Environmental Protection Agency. This report discusses the methods used to determine the minimum possible volume of the tank required to regulate wastewater discharge. With this information, it was determined that the use of a stirring tank is an efficient and cost effective way to regulate ammonia discharge. Based on these results many other companies may use this method to decrease the negative effects of ammonia on the environment.

  7. Synthesis of ammonia using sodium melt.

    Science.gov (United States)

    Kawamura, Fumio; Taniguchi, Takashi

    2017-09-14

    Research into inexpensive ammonia synthesis has increased recently because ammonia can be used as a hydrogen carrier or as a next generation fuel which does not emit CO 2 . Furthermore, improving the efficiency of ammonia synthesis is necessary, because current synthesis methods emit significant amounts of CO 2 . To achieve these goals, catalysts that can effectively reduce the synthesis temperature and pressure, relative to those required in the Haber-Bosch process, are required. Although several catalysts and novel ammonia synthesis methods have been developed previously, expensive materials or low conversion efficiency have prevented the displacement of the Haber-Bosch process. Herein, we present novel ammonia synthesis route using a Na-melt as a catalyst. Using this route, ammonia can be synthesized using a simple process in which H 2 -N 2 mixed gas passes through the Na-melt at 500-590 °C under atmospheric pressure. Nitrogen molecules dissociated by reaction with sodium then react with hydrogen, resulting in the formation of ammonia. Because of the high catalytic efficiency and low-cost of this molten-Na catalyst, it provides new opportunities for the inexpensive synthesis of ammonia and the utilization of ammonia as an energy carrier and next generation fuel.

  8. Characterization of an Alteromonas long-type ulvan lyase involved in the degradation of ulvan extracted from Ulva ohnoi.

    Science.gov (United States)

    He, Chuan; Muramatsu, Hisashi; Kato, Shin-Ichiro; Ohnishi, Kouhei

    2017-11-01

    Ulvan is a sulfated polysaccharide found in the cell wall of the green algae Ulva. We first isolated several ulvan-utilizing Alteromonas sp. from the feces of small marine animals. The strain with the highest ulvan-degrading activity, KUL17, was analyzed further. We identified a 55-kDa ulvan-degrading protein secreted by this strain and cloned the gene encoding for it. The deduced amino acid sequence indicated that the enzyme belongs to polysaccharide lyase family 24 and thus the protein was named ulvan lyase. The predicted molecular mass of this enzyme is 110 kDa, which is different from that of the identified protein. By deletion analysis, the catalytic domain was proven to be located on the N-terminal half of the protein. KUL17 contains two ulvan lyases, one long and one short, but the secreted and cleaved long ulvan lyase was demonstrated to be the major enzyme for ulvan degradation.

  9. [Construction and high-density fermentation of alkaline pectate lyase high-yield yeast].

    Science.gov (United States)

    Wang, Xiaowen; Xiang, La; Xu, Ting; Lu, Zhenghui; Zhang, Guimin

    2017-12-25

    Pectate lyase is widely applied in ramie degumming and fabric bioscouring in the textile industry. Compared to conventional processes that involve high alkaline and high temperature treatment, enzyme based treatments have significant advantages in fibers protectiveness, improved efficiency of refining, reduced energy consumption and pollution. Hence, it would be highly desirable to construct high-yield alkaline pectate lyase engineered strains and reduce the pectate lyase production cost. In the previous study, pectate lyase gene pel from Bacillus subtilis168 was expressed in Pichia pastoris GS115 after codon usage optimization based on the vector pHBM905A. To improve the expression level, the vector pHBM905BDM with optimized promoter and signal peptide was used to express the optimized gene pels in GS115. The transformant had increased activity from 68 U/mL to 100 U/mL with the improvement in the transcription level by 27% measured by qPCR. The transformants were further screened on pectin plates, where higher halo forming strains were picked for shake-flask fermentation and strain GS115-pHBM905BDM-pels4 showed the highest activity of 536 U/mL. Then plasmid pPIC9K-pels was constructed and electroporated into the GS115-pHBM905BDM-pels4 cells. Subsequently, high-copy transformant was screened by using the medium containing antibiotics G418, strain GS115-pHBM905BDMpPIC9K- pels1 was identified with increased activity of 770 U/mL and the copy number of pels was 7 confirmed by qPCR. Finally, the activity of pectate lyase produced by GS115-pHBM905BDM-pPIC9K-pels1reached to 2 271 U/mL in a 5-L fermentor. The activity of pectate lyase in our study reached the highest level of expression in P. pastoris, showing good application potential in the textile industry.

  10. Method for releasing hydrogen from ammonia borane

    Science.gov (United States)

    Varma, Arvind; Diwan, Moiz; Shafirovich, Evgeny; Hwang, Hyun-Tae; Al-Kukhun, Ahmad

    2013-02-19

    A method of releasing hydrogen from ammonia borane is disclosed. The method comprises heating an aqueous ammonia borane solution to between about 80-135.degree. C. at between about 14.7 and 200 pounds per square inch absolute (psia) to release hydrogen by hydrothermolysis.

  11. Chilled ammonia process for CO2 capture

    DEFF Research Database (Denmark)

    Darde, Victor Camille Alfred; Thomsen, Kaj; van Well, Willy J. M

    2009-01-01

    The chilled ammonia process absorbs the CO2 at low temperature (2-10 degrees C). The heat of absorption of carbon dioxide by ammonia is significantly lower than for amines. In addition, degradation problems can be avoided and a high carbon dioxide capacity is achieved. Hence, this process shows...

  12. Regeneration of ammonia borane from polyborazylene

    Science.gov (United States)

    Sutton, Andrew; Gordon, John C; Ott, Kevin C; Burrell, Anthony K

    2013-02-05

    Method of producing ammonia borane, comprising providing a reagent comprising a dehydrogenated material in a suitable solvent; and combining the reagent with a reducing agent comprising hydrazine, a hydrazine derivative, or combinations thereof, in a reaction which produces a mixture comprising ammonia borane.

  13. Chilled Ammonia Process for CO2 Capture

    DEFF Research Database (Denmark)

    Darde, Victor Camille Alfred; Thomsen, Kaj; Well, Willy J.M. van

    2010-01-01

    The chilled ammonia process absorbs the CO2 at low temperature (2–10°C). The heat of absorption of carbon dioxide by ammonia is significantly lower than for amines. In addition, degradation problems can be avoided and a high carbon dioxide capacity is achieved. Hence, this process shows good...

  14. Atmospheric behaviour of ammonia and ammonium

    NARCIS (Netherlands)

    Asman, W.A.H.

    1987-01-01

    1.4.1 Scope of this thesis

    A few models for ammonia and ammonium exist. Russell et al. (1983) made a multi-layer Lagrangian transport model describing the transport and formation of ammonium nitrate aerosol for California. They did not take reactions of ammonia and sulphuric acid

  15. Reducing ammonia volatilization from compound fertilizers ...

    African Journals Online (AJOL)

    Ammonia volatilization is a direct loss of available nitrogen in agriculture. The objective of this study was to determine the effect of amending NPK fertilizer with different rates of clinoptilolite zeolite on ammonia volatilization, soil exchangeable ammonium, and available nitrate. Seven treatments evaluated were: 250 g soil ...

  16. Ammonia Sensor Using Wavelength Modulation Spectroscopy

    KAUST Repository

    Farooq, Aamir

    2015-09-01

    An ammonia sensor can include a laser detector configured to provide stable sample readings. The sensor can implement a method including processing the recorded intensity of the laser beam to determine a first harmonic component and a second harmonic component and the amount of ammonia in the sample.

  17. Ammonia Solubility in High Concentration Salt Solutions

    International Nuclear Information System (INIS)

    HEDENGREN, D.C.

    2000-01-01

    Solubility data for ammonia in water and various dilute solutions are abundant in the literature. However, there is a noticeable lack of ammonia solubility data for high salt, basic solutions of various mixtures of salts including those found in many of the Hanford Washington underground waste tanks. As a result, models based on solubility data for dilute salt solutions have been used to extrapolate to high salt solutions. These significant extrapolations need to be checked against actual laboratory data. Some indirect vapor measurements have been made. A more direct approach is to determine the ratio of solubility of ammonia in water to its solubility in high salt solutions. In various experiments, pairs of solutions, one of which is water and the other a high salt solution, are allowed to come to equilibrium with a common ammonia vapor pressure. The ratio of concentrations of ammonia in the two solutions is equal to the ratio of the respective ammonia solubilities (Henry's Law constants) at a given temperature. This information can then be used to refine the models that predict vapor space compositions of ammonia. Ammonia at Hanford is of concern because of its toxicity in the environment and its contribution to the flammability of vapor space gas mixtures in waste tanks

  18. Ammonia Concentrations in Different Aquaculture Holding Tanks ...

    African Journals Online (AJOL)

    High unionized ammonia recorded in the collapsible and concrete ponds was from excretion of high protein rich feed, decomposition of uneaten feed, high stocking density, low water exchange rates, water source and the alkaline medium of the systems. Low unionized ammonia in earthen pond and natural pond was ...

  19. Measuring ammonia emissions from manured fields

    NARCIS (Netherlands)

    Berkhout, A.J.C.; Hoff, G.R.; Bergwerff, J.B.; Swart, D.P.J.; Hensen, A.; Kraai, A.; Bleeker, A.; Huijsmans, J.F.M.; Mosquera Losada, J.; Pul, van W.A.J.

    2008-01-01

    In this report, 2 novel instruments are described that are able to measure the ammonia emissions of manured fields. The 1st instrument, developed and operated by ECN, is a tuneable diode laser spectrometer (TDL), mounted in a van. It is used to measure the ammonia concentration patterns downwind

  20. New findings on cerebral ammonia uptake in HE using functional (13)N-ammonia PET.

    Science.gov (United States)

    Sørensen, Michael; Keiding, Susanne

    2007-12-01

    PET is a functional imaging technique suitable for studies of brain ammonia metabolism. Dynamic (13)N-ammonia PET yields time-courses of radioactivity concentrations in brain (PET camera) and blood (samples). Ahl et al. (Hepatology 40:73-79, 2004) and Keiding et al. (Hepatology 43:42-50, 2006) analysed such data in patients with HE by a kinetic model accounting for transfer of (13)N-ammonia across the blood-brain barrier (BBB) and intracellular formation of (13)N-glutamine. Initial unidirectional (13)N-ammonia transfer across BBB was characterized by the permeability-surface area product PS(BBB) (ml blood min(-1) ml(-1) tissue). There was a tendency to lower PS(BBB) values in patients with cirrhosis and HE than in patients with cirrhosis without HE and healthy controls but the differences were not statistically significant. Keiding et al. (Hepatology 43:42-50, 2006) also calculated PS(met) (ml blood min(-1) ml(-1) tissue) as a measure of the combined transfer of (13)N-ammonia across BBB and subsequent intracellular metabolism of (13)N-ammonia; neither did this PS-value show significant difference between the groups of subjects. Net flux of ammonia from blood into intracellular metabolites was linearly correlated to arterial ammonia. In conclusion, basic brain ammonia kinetics was not changed significantly in patients with cirrhosis +/- HE compared to healthy controls. Blood ammonia seems to be the more important factor for increased brain ammonia uptake in HE.

  1. Enrichment of ammonia concentration from aqua-ammonia vapors by using 3A molecular sieve

    International Nuclear Information System (INIS)

    Chiou, J.S.; Lin, T.M.; She, K.Y.; Chen, W.M.

    2009-01-01

    In aqua-ammonia refrigeration systems, the ammonia is the refrigerant and the water is the absorbent, the vapor produced in the generator always contains a small fraction of water. The removed of this residual water is a crucial issue in order to guarantee a reliable and efficient operation of these systems. Currently, the thermal distillation methods (via a rectifier and/or an analyzer) are used to further separate the water from aqua-ammonia mixtures. In this study, a molecular sieve module is used for ammonia purification. A thermal system with a 3A molecular sieve module was set up, and the conditions of working fluid entering into the sieve module is similar to that entering into the rectifier tower of a typical aqua-ammonia absorption system. Results from ammonia enrichment tests indicate the concentration of ammonia can be raised from about 80% up to about 99% if siever installation was properly arranged.

  2. Ammonia and urea permeability of mammalian aquaporins

    DEFF Research Database (Denmark)

    Litman, Thomas; Søgaard, Rikke; Zeuthen, Thomas

    2009-01-01

    and 9 are found together with Rh proteins in cells exposed to portal blood coming from the intestine. In the kidney, AQP3 might participate in the excretion of NH(4) (+) in the collecting duct. The interplay between the ammonia-permeable aquaporins and the other types of ammonia- and urea......The human aquaporins,AQP3,AQP7, AQP8,AQP9, and possibly AQP10, are permeable to ammonia, and AQP7, AQP9, and possibly AQP3, are permeable to urea. In humans, these aquaporins supplement the ammonia transport of the Rhesus (Rh) proteins and the urea transporters (UTs). The mechanism by which...... ammonium is transported by aquaporins is not fully resolved. A comparison of transport equations, models, and experimental data shows that ammonia is transported in its neutral form, NH(3). In the presence of NH(3), the aquaporin stimulates H(+) transport. Consequently, this transport of H(+) is only...

  3. Ammonia transformation in a biotrickling air filter

    DEFF Research Database (Denmark)

    Nielsen, Lars Peter; Nielsen, Marie Louise; Andersen, Mathias

    2007-01-01

    A simple, tubular biotrickling filter was designed for optimal removal of ammonia and odour in ventilation air from a pig house. The removal and transformation of ammonia was studied in detail by analysis and modelling of chemical gradients through the filter. Good correspondence between...... measurements and model was obtained by using conventional substrate and inhibition kinetics of ammonium and nitrite oxidizing bacteria. Highest rates of ammonia removal were observed in the central section of the filter. Near the air outlet and water inlet the process was ammonia limited, while high nitrous...... acid concentrations almost excluded any biological activity near the air inlet and water outlet. Nitrous acid inhibition also stabilized pH at 6.5-7 all through the filter. Being sensitive to both ammonia and nitrous acid the nitrite oxidation process occurred mainly in the filter sections near the air...

  4. Crystallization and preliminary X-ray analysis of argininosuccinate lyase from Streptococcus mutans

    International Nuclear Information System (INIS)

    Cao, Yan-Li; Li, Gui-Lan; Wang, Kai-Tuo; Zhang, Hong-Yin; Li, Lan-Fen

    2011-01-01

    Crystals of argininosuccinate lyase from S. mutans were obtained and X-ray data were collected to 2.5 Å resolution in space group R3. Argininosuccinate lyase (ASL) is an important enzyme in arginine synthesis and the urea cycle, which are highly conserved from bacteria to eukaryotes. The gene encoding Streptococcus mutans ASL (smASL) was amplified and cloned into expression vector pET28a. The recombinant smASL protein was expressed in a soluble form in Escherichia coli strain BL21 (DE3) and purified to homogeneity by two-step column chromatography. Crystals suitable for X-ray analysis were obtained and X-ray diffraction data were collected to a resolution of 2.5 Å. The crystals belonged to space group R3, with unit-cell parameters a = b = 254.5, c = 78.3 Å

  5. New lupane triterpenoids from Solidago canadensis that inhibit the lyase activity of DNA polymerase beta.

    Science.gov (United States)

    Chaturvedula, V S Prakash; Zhou, Bing-Nan; Gao, Zhijie; Thomas, Shannon J; Hecht, Sidney M; Kingston, David G I

    2004-12-01

    Bioassay-directed fractionation of a methyl ethyl ketone extract of Solidago canadensis L. (Asteraceae), using an assay to detect the lyase activity of DNA polymerase beta, resulted in the isolation of the four new lupane triterpenoids 1-4 and the seven known compounds lupeol, lupeyl acetate, ursolic acid, cycloartenol, cycloartenyl palmitate, alpha-amyrin acetate, and stigmasterol. The structures of the new compounds were established as 3beta-(3R-acetoxyhexadecanoyloxy)-lup-20(29)-ene (1), 3beta-(3-ketohexadecanoyloxy)-lup-20(29)-ene (2), 3beta-(3R-acetoxyhexadecanoyloxy)-29-nor-lupan-20-one (3), and 3beta-(3-hetohexadecanoyloxy)-29-nor-lupan-20-one (4), respectively, on the basis of extensive 1D and 2D NMR spectroscopic interpretation and chemical modification studies. All 11 compounds were inhibitory to the lyase activity of DNA polymerase beta.

  6. Mechanism of Cytochrome P450 17A1-Catalyzed Hydroxylase and Lyase Reactions

    DEFF Research Database (Denmark)

    Bonomo, Silvia; Jorgensen, Flemming Steen; Olsen, Lars

    2017-01-01

    Cytochrome P450 17A1 (CYP17A1) catalyzes C17 hydroxylation of pregnenolone and progesterone and the subsequent C17–C20 bond cleavage (lyase reaction) to form androgen precursors. Compound I (Cpd I) and peroxo anion (POA) are the heme-reactive species underlying the two reactions. We have...... states (TSs) for the two reactions into the active site of CYP17A1 showed that the TS for the C17 hydroxylation needs to be distorted by 13 kJ·mol–1, whereas the TS for the 17,20 lyase reaction easily can be accommodated in the protein. Finally, differences in the hydrogen-bond pattern of the substrates...

  7. Abundance and genetic diversity of microbial polygalacturonase and pectate lyase in the sheep rumen ecosystem.

    Science.gov (United States)

    Yuan, Peng; Meng, Kun; Wang, Yaru; Luo, Huiying; Huang, Huoqing; Shi, Pengjun; Bai, Yingguo; Yang, Peilong; Yao, Bin

    2012-01-01

    Efficient degradation of pectin in the rumen is necessary for plant-based feed utilization. The objective of this study was to characterize the diversity, abundance, and functions of pectinases from microorganisms in the sheep rumen. A total of 103 unique fragments of polygalacturonase (PF00295) and pectate lyase (PF00544 and PF09492) genes were retrieved from microbial DNA in the rumen of a Small Tail Han sheep, and 66% of the sequences of these fragments had low identities (pectinases. The two most abundant pectate lyase genes were cloned, and their protein products, expressed in Escherichia coli, were characterized. Both enzymes probably act extracellularly as their nucleotide sequences contained signal sequences, and they had optimal activities at the ruminal physiological temperature and complementary pH-dependent activity profiles. This study reveals the specificity, diversity, and abundance of pectinases in the rumen ecosystem and provides two additional ruminal pectinases for potential industrial use under physiological conditions.

  8. Biochemical Stability and Molecular Dynamic Characterization of Aspergillus fumigatus Cystathionine γ-Lyase in Response to Various Reaction Effectors

    KAUST Repository

    El-Sayed, Ashraf S.A.

    2015-08-11

    Cystathionine γ-lyase (CGL) is a key enzyme in the methionine-cysteine cycle in all living organisms forming cysteine, α-ketobutyrate and ammonia via homocysteine and cystathionine intermediates. Although, human and plant CGLs have been extensively studied at the molecular and mechanistic levels, there has been little work on the molecular and catalytic properties of fungal CGL. Herein, we studied in detail for the first time the molecular and catalytic stability of Aspergillus fumigatus CGL, since conformational instability, inactivation and structural antigenicity are the main limitations of the PLP-dependent enzymes on various therapeutic uses. We examined these properties in response to buffer compositions, stabilizing and destabilizing agents using Differential Scanning Fluorometery (DSF), steady state and gel-based fluorescence of the intrinsic hydrophobic core, stability of internal aldimine linkage and catalytic properties. The activity of the recombinant A. fumigatus CGL was 13.8 U/mg. The melting temperature (Tm) of CGL in potassium phosphate buffer (pH 7.0-8.0) was 73.3 °C, with ∼3 °C upshifting in MES and sodium phosphate buffers (pH 7.0). The conformational thermal stability was increased in potassium phosphate, sodium phosphate and MES buffers, in contrast to Tris-HCl, HEPES (pH 7.0) and CAPS (pH 9.0-10.0). The thermal stability and activity of CGL was slightly increased in the presence of trehalose and glycerol that might be due to hydration of the enzyme backbone, unlike the denaturing effect of GdmCl and urea. Modification of surface CGL glutamic and aspartic acids had no significant effect on the enzyme conformational and catalytic stability. Molecular modeling and dynamics simulations unveil the high conformational stability of the overall scaffold of CGL with high flexibility at the non-structural regions. CGL structure has eight buried Trp residues, which are reoriented to the enzyme surface and get exposed to the solvent under

  9. Clinical utility of breath ammonia for evaluation of ammonia physiology in healthy and cirrhotic adults

    Science.gov (United States)

    Spacek, Lisa A; Mudalel, Matthew; Tittel, Frank; Risby, Terence H; Solga, Steven F

    2016-01-01

    Blood ammonia is routinely used in clinical settings to assess systemic ammonia in hepatic encephalopathy and urea cycle disorders. Despite its drawbacks, blood measurement is often used as a comparator in breath studies because it is a standard clinical test. We sought to evaluate sources of measurement error and potential clinical utility of breath ammonia compared to blood ammonia. We measured breath ammonia in real time by quartz enhanced photoacoustic spectrometry and blood ammonia in 10 healthy and 10 cirrhotic participants. Each participant contributed 5 breath samples and blood for ammonia measurement within 1 h. We calculated the coefficient of variation (CV) for 5 breath ammonia values, reported medians of healthy and cirrhotic participants, and used scatterplots to display breath and blood ammonia. For healthy participants, mean age was 22 years (±4), 70% were men, and body mass index (BMI) was 27 (±5). For cirrhotic participants, mean age was 61 years (±8), 60% were men, and BMI was 31 (±7). Median blood ammonia for healthy participants was within normal range, 10 μmol L−1 (interquartile range (IQR), 3–18) versus 46 μmol L−1 (IQR, 23–66) for cirrhotic participants. Median breath ammonia was 379 pmol mL−1 CO2 (IQR, 265–765) for healthy versus 350 pmol mL−1 CO2 (IQR, 180–1013) for cirrhotic participants. CV was 17 ± 6%. There remains an important unmet need in the evaluation of systemic ammonia, and breath measurement continues to demonstrate promise to fulfill this need. Given the many differences between breath and blood ammonia measurement, we examined biological explanations for our findings in healthy and cirrhotic participants. We conclude that based upon these preliminary data breath may offer clinically important information this is not provided by blood ammonia. PMID:26658550

  10. Mandelonitrile lyase from Ximenia americana L.: stereospecificity and lack of flavin prosthetic group.

    Science.gov (United States)

    Kuroki, G W; Conn, E E

    1989-09-01

    A mandelonitrile lyase (EC 4.1.2.10) that catalyzes the dissociation of (S)-(-)-mandelonitrile to benzaldehyde and hydrogen cyanide has been purified to apparent homogeneity from leaves of Ximenia americana L. (Olacaceae). The lyase was purified 122-fold with 38% yield by chromatography on carboxymethyl-cellulose and chromatofocusing. The enzyme had a pH optimum of 5.5, with a Km value of 280 microM. Activity toward 4-hydroxy-(R,S)-mandelonitrile was 77% of that observed with the endogenous substrate; no activity was observed toward the aliphatic substrate acetone cyanohydrin. The enzyme was stable at 4 degrees C and at room temperature for at least 1 month. Native and subunit molecular weights of 38,000 and 36,500, respectively, suggest the enzyme is a monomer. The isoelectric point was pH 3.9 as determined by isoelectric focusing. Staining with periodic acid-Schiff and fluorescein-labeled concanavalin A reagents indicate this enzyme is a glycoprotein. In contrast to (R)-mandelonitrile lyases isolated from Prunus species, the Ximenia lyase does not appear to be a flavoprotein. A second enzyme that eluted from the chromatofocusing column at pH 4.0 was also active toward mandelonitrile. However, this form accounted for less than 10% of the total activity, and its specific activity was only 6% of that of the major component. Additional physical and kinetic studies suggested this activity may be due to a nonspecific enzyme that is active toward mandelonitrile.

  11. Progress Towards Simple and Direct Detection of Adenylosuccinate Lyase Deficiency in Human Urine.

    Science.gov (United States)

    Lim, Soojin; Lowry, Mark; Strongin, Robert M

    2011-10-14

    A rhodamine based boronic acid linearly responds to increasing 5-aminoimidazole-4-carboxamide riboside (AICAr) concentrations in human urine. This method is thus an advance in detecting adenylosuccinate lyase (ADSL) deficiency as AICAr is a model riboside for the ADSL substrates succinyladenosine (S-Ado) and succinylaminoimidazolecarboxamide riboside (SAICAr). ADSL deficiency is a rare but devastating disease of de novo purine synthesis in infants. Its diagnosis is also significant as it is one of the autism spectrum disorders.

  12. Improvement of aromatic thiol release through the selection of yeasts with increased β-lyase activity.

    Science.gov (United States)

    Belda, Ignacio; Ruiz, Javier; Navascués, Eva; Marquina, Domingo; Santos, Antonio

    2016-05-16

    The development of a selective medium for the rapid differentiation of yeast species with increased aromatic thiol release activity has been achieved. The selective medium was based on the addition of S-methyl-l-cysteine (SMC) as β-lyase substrate. In this study, a panel of 245 strains of Saccharomyces cerevisiae strains was tested for their ability to grow on YCB-SMC medium. Yeast strains with an increased β-lyase activity grew rapidly because of their ability to release ammonium from SMC in comparison to others, and allowed for the easy isolation and differentiation of yeasts with promising properties in oenology, or another field, for aromatic thiol release. The selective medium was also helpful for the discrimination between those S. cerevisiae strains, which present a common 38-bp deletion in the IRC7 sequence (present in around 88% of the wild strains tested and are likely to be less functional for 4-mercapto-4-methylpentan-2-one (4MMP) production), and those S. cerevisiae strains homozygous for the full-length IRC7 allele. The medium was also helpful for the selection of non-Saccharomyces yeasts with increased β-lyase activity. Based on the same medium, a highly sensitive, reproducible and non-expensive GC-MS method for the evaluation of the potential volatile thiol release by different yeast isolates was developed. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Characterization of a Long-Lived Alginate Lyase Derived from Shewanella Species YH1

    Directory of Open Access Journals (Sweden)

    Hisashi Yagi

    2017-12-01

    Full Text Available Polysaccharides from seaweeds are widely used in various fields, including the food, biomedical material, cosmetic, and biofuel industries. Alginate, which is a major polysaccharide in brown algae, and the products of its degradation (oligosaccharides have been used in stabilizers, thickeners, and gelling agents, especially in the food industry. Discovering novel alginate lyases with unique characteristics for the efficient production of oligosaccharides may be relevant for the food and pharmaceutical fields. In this study, we identified a unique alginate lyase derived from an alginate-utilizing bacterium, Shewanella species YH1. The recombinant enzyme (rAlgSV1-PL7 was produced in an Escherichia coli system and it was classified in the Polysaccharide Lyase family 7. The optimal temperature and pH for rAlgSV1-PL7 activity were around 45 °C and 8, respectively. Interestingly, we observed that rAlgSV1-PL7 retained over 80% of its enzyme activity after incubation at 30 °C for at least 20 days, indicating that rAlgSV1-PL7 is a long-lived enzyme. Moreover, the degradation of alginate by rAlgSV1-PL7 produced one to four sugars because of the broad substrate specificity of this enzyme. Our findings suggest that rAlgSV1-PL7 may represent a new commercially useful enzyme.

  14. Characterization of a Long-Lived Alginate Lyase Derived from Shewanella Species YH1.

    Science.gov (United States)

    Yagi, Hisashi; Isobe, Natsuki; Itabashi, Narumi; Fujise, Asako; Ohshiro, Takashi

    2017-12-27

    Polysaccharides from seaweeds are widely used in various fields, including the food, biomedical material, cosmetic, and biofuel industries. Alginate, which is a major polysaccharide in brown algae, and the products of its degradation (oligosaccharides) have been used in stabilizers, thickeners, and gelling agents, especially in the food industry. Discovering novel alginate lyases with unique characteristics for the efficient production of oligosaccharides may be relevant for the food and pharmaceutical fields. In this study, we identified a unique alginate lyase derived from an alginate-utilizing bacterium, Shewanella species YH1. The recombinant enzyme (rAlgSV1-PL7) was produced in an Escherichia coli system and it was classified in the Polysaccharide Lyase family 7. The optimal temperature and pH for rAlgSV1-PL7 activity were around 45 °C and 8, respectively. Interestingly, we observed that rAlgSV1-PL7 retained over 80% of its enzyme activity after incubation at 30 °C for at least 20 days, indicating that rAlgSV1-PL7 is a long-lived enzyme. Moreover, the degradation of alginate by rAlgSV1-PL7 produced one to four sugars because of the broad substrate specificity of this enzyme. Our findings suggest that rAlgSV1-PL7 may represent a new commercially useful enzyme.

  15. Cloning and study of the pectate lyase gene of Erwinia carotovora

    International Nuclear Information System (INIS)

    Bukanov, N.O.; Fonshtein, M.Yu.; Evtushenkov, A.N.; Syarinskii, M.A.; Strel'chenko, P.P.; Yankovski, N.K.; Alikhanyan, S.I.; Fomichev, Yu.K.; Debabov, V.G.

    1986-01-01

    The cloning of the gene of a secretable protein of Erwinia carotovora, pectate lyase, in Escherichia coli was described. Primary cloning was conducted using the phage vector λ 47.1. In the gene library of E. carotovora obtained, eight phages carrying the gene sought were identified according to the appearance of enzymatic activity of the gene product, pectate lyase, in situ. The BamHI fragment of DNA, common to all these phages, was recloned on the plasmid pUC19. It was shown that the cloned pectate lyase gene is represented on the E. carotovora chromosome in one copy. Methods of production of representative gene libraries on phage vectors from no less than 1 μg of cloned DNA even for the genomes of eukaryotes have now been developed. Vectors have been created, for example, λ 47.1, permitting the selection only of hybrid molecules. A number of methods have been developed for the search for a required gene in the library, depending on whether the cloned gene can be expressed or not, and if it can, what properties it will impart to the hybrid clone containing it

  16. Purification and Characterization of a Novel (R)-Mandelonitrile Lyase from the Fern Phlebodium aureum.

    Science.gov (United States)

    Wajant, H.; Forster, S.; Selmar, D.; Effenberger, F.; Pfizenmaier, K.

    1995-12-01

    Using high-performance liquid chromatography and nuclear magnetic resonance we identified vicianin as the cyanogenic compound of Phlebodium aureum. The (R)-hydroxynitrile lyase involved during cyanogenesis in the catabolism of the aglycon ([R]-mandelonitrile) was purified to apparent homogeneity. The purified holoenzyme is a homomultimer with subunits of Mr = 20,000. At least three isoforms of the enzyme exist. In contrast to other hydroxynitrile lyases, mandelonitrile lyase (MDL) from P. aureum was not inhibited by sulfhydryl- or hydroxyl-modifying reagents, suggesting a different catalytic mechanism. The enzyme is active over a broad temperature range, with maximum activity between 35 and 50[deg]C, and a pH optimum at 6.5. In contrast to (R)-MDLs isolated from several species of the Rosaceae family, (R)-MDL from P. aureum is not a flavoprotein. The substrate specificity was investigated using immobilized enzyme and diisopropyl ether as solvent. The addition of cyanide to aromatic and heterocyclic carbonyls is catalyzed by this (R)-MDL, whereas aliphatic carbonyls are poorly converted.

  17. Neurologic presentation, diagnostics, and therapeutic insights in a severe case of adenylosuccinate lyase deficiency.

    Science.gov (United States)

    Jurecka, Agnieszka; Opoka-Winiarska, Violetta; Rokicki, Dariusz; Tylki-Szymańska, Anna

    2012-05-01

    Epilepsy in adenylosuccinate lyase deficiency may be difficult to treat, and there is no standardized therapy. The authors describe a case of severe adenylosuccinate lyase deficiency resulting from a heterozygous mutation of the ADSL gene (p.D215H/p.I351T). The patient presented with tonic-clonic seizures, opisthotonus, tremor, and myoclonus in the 4th day of life. The seizures were refractory on various combinations of antiepileptic treatment. A ketogenic diet was introduced at the age of 2 resulting in a seizure-free period. The patient, however, developed a metabolic hyperchloremic acidosis with Fanconi syndrome, which disappeared a month after cessation of the diet at the age of 5. Since the withdrawal of the ketogenic diet, seizures have returned to a frequency of several times a day. In conclusion, a ketogenic diet could be considered a valid therapeutic option in patients with intractable seizures in a course of adenylosuccinate lyase deficiency; however, it requires a formal study.

  18. Vapor Phase Catalytic Ammonia Reduction

    Science.gov (United States)

    Flynn, Michael T.; Harper, Lynn D. (Technical Monitor)

    1994-01-01

    This paper discusses the development of a Vapor Phase Catalytic Ammonia Reduction (VPCAR) teststand and the results of an experimental program designed to evaluate the potential of the technology as a water purification process. In the experimental program the technology is evaluated based upon product water purity, water recovery rate, and power consumption. The experimental work demonstrates that the technology produces high purity product water and attains high water recovery rates at a relatively high specific power consumption. The experimental program was conducted in 3 phases. In phase I an Igepon(TM) soap and water mixture was used to evaluate the performance of an innovative Wiped-Film Rotating-Disk evaporator and associated demister. In phase II a phenol-water solution was used to evaluate the performance of the high temperature catalytic oxidation reactor. In phase III a urine analog was used to evaluate the performance of the combined distillation/oxidation functions of the processor.

  19. Molecular cloning, purification, and characterization of a novel polyMG-specific alginate lyase responsible for alginate MG block degradation in Stenotrophomas maltophilia KJ-2

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Su In; Kim, Hee Sook [Kyungsung Univ., Busan (Korea, Republic of). Dept. of Food Science and Biotechnology; Choi, Sung Hee; Lee, Eun Yeol [Kyung Hee Univ., Gyeonggi-do (Korea, Republic of). Dept. of Chemical Engineering

    2012-09-15

    A gene for a polyMG-specific alginate lyase possessing a novel structure was identified and cloned from Stenotrophomas maltophilia KJ-2 by using PCR with homologous nucleotide sequences-based primers. The recombinant alginate lyase consisting of 475 amino acids was purified on Ni-Sepharose column and exhibited the highest activity at pH 8 and 40 C. Interestingly, the recombinant alginate lyase was expected to have a similar catalytic active site of chondroitin B lyase but did not show chondroitin lyase activity. In the test of substrate specificity, the recombinant alginate lyase preferentially degraded the glycosidic bond of polyMG-block than polyM-block and polyG-block. The chemical structures of the degraded alginate oligosaccharides were elucidated to have mannuronate (M) at the reducing end on the basis of NMR analysis, supporting that KJ-2 polyMG-specific alginate lyase preferably degraded the glycosidic bond in M-G linkage than that in G-M linkage. The KJ-2 polyMG-specific alginate lyase can be used in combination with other alginate lyases for a synergistic saccharification of alginate. (orig.)

  20. IRIS Toxicological Review of Ammonia (Revised External ...

    Science.gov (United States)

    In August 2013, EPA submitted a revised draft IRIS assessment of ammonia to the agency's Science Advisory Board (SAB) and posted this draft on the IRIS website. EPA had previously released a draft of the assessment for public comment, held a public meeting about the draft, and then revised it based on the comments received. The SAB CAAC-Ammonia panel will review this draft assessment. Details about the meeting dates, times, and location are available via the Federal Register Notice posted on March 25, 2014. The SAB provided information on how the public can participate in the external peer review meetings, as well as instructions about how to provide comments to the SAB in the notice. Additional information on the SAB review of ammonia is on the SAB website. Report Information: The Toxicological Review of Ammonia was originally released for a 60-day public comment period on June 8, 2012. [Federal Register Notice Jun 8, 2012] EPA revised the toxicological review in response to the public comments received. EPA has released the revised external review draft ammonia assessment and the SAB CAAC is conducting a peer review of the scientific basis supporting the assessment that will appear in the Integrated Risk Information System (IRIS) database. Information regarding the peer review can be found at the SAB review of ammonia website. EPA is undertaking an Integrated Risk Information System (IRIS) health assessment for ammonia. IRIS is an EPA database cont

  1. Resveratrol Prevents Ammonia Toxicity in Astroglial Cells

    Science.gov (United States)

    Guerra, Maria Cristina; Leite, Marina Concli; Souza, Diogo Onofre; Gonçalves, Carlos-Alberto; Gottfried, Carmem

    2012-01-01

    Ammonia is implicated as a neurotoxin in brain metabolic disorders associated with hyperammonemia. Acute ammonia toxicity can be mediated by an excitotoxic mechanism, oxidative stress and nitric oxide (NO) production. Astrocytes interact with neurons, providing metabolic support and protecting against oxidative stress and excitotoxicity. Astrocytes also convert excess ammonia and glutamate into glutamine via glutamine synthetase (GS). Resveratrol, a polyphenol found in grapes and red wines, exhibits antioxidant and anti-inflammatory properties and modulates glial functions, such as glutamate metabolism. We investigated the effect of resveratrol on the production of reactive oxygen species (ROS), GS activity, S100B secretion, TNF-α, IL-1β and IL-6 levels in astroglial cells exposed to ammonia. Ammonia induced oxidative stress, decreased GS activity and increased cytokines release, probably by a mechanism dependent on protein kinase A (PKA) and extracellular signal-regulated kinase (ERK) pathways. Resveratrol prevented ammonia toxicity by modulating oxidative stress, glial and inflammatory responses. The ERK and nuclear factor-κB (NF-κB) are involved in the protective effect of resveratrol on cytokines proinflammatory release. In contrast, other antioxidants (e.g., ascorbic acid and trolox) were not effective against hyperammonemia. Thus, resveratrol could be used to protect against ammonia-induced neurotoxicity. PMID:23284918

  2. Resveratrol prevents ammonia toxicity in astroglial cells.

    Directory of Open Access Journals (Sweden)

    Larissa Daniele Bobermin

    Full Text Available Ammonia is implicated as a neurotoxin in brain metabolic disorders associated with hyperammonemia. Acute ammonia toxicity can be mediated by an excitotoxic mechanism, oxidative stress and nitric oxide (NO production. Astrocytes interact with neurons, providing metabolic support and protecting against oxidative stress and excitotoxicity. Astrocytes also convert excess ammonia and glutamate into glutamine via glutamine synthetase (GS. Resveratrol, a polyphenol found in grapes and red wines, exhibits antioxidant and anti-inflammatory properties and modulates glial functions, such as glutamate metabolism. We investigated the effect of resveratrol on the production of reactive oxygen species (ROS, GS activity, S100B secretion, TNF-α, IL-1β and IL-6 levels in astroglial cells exposed to ammonia. Ammonia induced oxidative stress, decreased GS activity and increased cytokines release, probably by a mechanism dependent on protein kinase A (PKA and extracellular signal-regulated kinase (ERK pathways. Resveratrol prevented ammonia toxicity by modulating oxidative stress, glial and inflammatory responses. The ERK and nuclear factor-κB (NF-κB are involved in the protective effect of resveratrol on cytokines proinflammatory release. In contrast, other antioxidants (e.g., ascorbic acid and trolox were not effective against hyperammonemia. Thus, resveratrol could be used to protect against ammonia-induced neurotoxicity.

  3. Regulation of acetate metabolism in Corynebacterium glutamicum: transcriptional control of the isocitrate lyase and malate synthase genes.

    Science.gov (United States)

    Wendisch, V F; Spies, M; Reinscheid, D J; Schnicke, S; Sahm, H; Eikmanns, B J

    1997-10-01

    In the amino-acid-producing microorganism Corynebacterium glutamicum, the specific activities of the acetate-activating enzymes acetate kinase and phosphotransacetylase and those of the glyoxylate cycle enzymes isocitrate lyase and malate synthase were found to be high when the cells were grown on acetate (0.8, 2.9, 2.1, and 1.8 U/mg protein, respectively). When the cells were grown on glucose or on other carbon sources such as lactate, succinate, or glutamate, the specific activities were two- to fourfold (acetate kinase and phosphotransacetylase) and 45- to 100-fold (isocitrate lyase and malate synthase) lower, indicating that the synthesis of the four enzymes is regulated by acetate in the growth medium. A comparative Northern (RNA) analysis of the C. glutamicum isocitrate lyase and malate synthase genes (aceA and aceB) and transcriptional cat fusion experiments revealed that aceA and aceB are transcribed as 1.6- and 2.7-kb monocistronic messages, respectively, and that the regulation of isocitrate lyase and malate synthase synthesis is exerted at the level of transcription from the respective promoters. Surprisingly, C. glutamicum mutants defective in either acetate kinase or phosphotransacetylase showed low specific activities of the other three enzymes (phosphotransacetylase, isocitrate lyase, and malate synthase or acetate kinase, isocitrate lyase, and malate synthase, respectively) irrespective of the presence or absence of acetate in the medium. This result and a correlation of a high intracellular acetyl coenzyme A concentration with high specific activities of isocitrate lyase, malate synthase, acetate kinase, and phosphotransacetylase suggest that acetyl coenzyme A or a derivative thereof may be a physiological trigger for the genetic regulation of enzymes involved in acetate metabolism of C. glutamicum.

  4. Transformation of ammonia i biological airfilters

    DEFF Research Database (Denmark)

    Nielsen, Lars Peter; Sørensen, Karen; Andersen, Mathias

    2007-01-01

    Ammonia is a major compound in ventilation air from animal houses. In biological filters it is with varying efficiency transformed by physical, biological, and chemical processes and ends up as ammonium, nitrate, and nitrite dissolved in water and as dinitrogen, nitrous oxide and nitric oxide...... nitrite oxidation only appeared in locations with minimal ammonia and nitrite levels. Nitrous oxide emission depended on anoxic microsites, and nitric oxide production was associated with nitrite accumulation. Water and biomass management appear to be the important tools for optimization of ammonia...

  5. Electrochemical monitoring of ammonia during anaerobic digestion

    DEFF Research Database (Denmark)

    Zhao, Nannan; Angelidaki, Irini; Zhang, Yifeng

    Ammonia is known as key inhibitor to methanogens in anaerobic digestion (AD) process. It’s of importance to develop efficient tool for ammonia monitoring. In this study, an electrolysis cell (EC) coupled with a complete nitrification reactor was developed as sensor for real time and online......-rich digesters. It was observed that the initial transient currents (0 min) were linearly corresponding to the ammonia levels (from 0 to 95.75 mg/L NH4+-N, R2 = 0.9673). Finally, this new sensor was tested with real AD effluent and the results showed no significant difference with that measured by conventional...

  6. Ammonia complexes of metals in aqueous solutions with high concentrations of ammonia

    International Nuclear Information System (INIS)

    Padar, T.G.; Novikov, L.K.; Stupko, T.V.; Isaev, I.D.; Pashkov, G.L.; Mironov, V.E.

    1991-01-01

    Potentiometric method, glass electrodes and Bierrum function were used to study the formation of ammonia complexes of magnesium, calcium, cadmium, zinc, copper(2) and silver in 2.0 mol/dm 3 aqueous solutions of ammonia nitrate with 0-18 mol/dm 3 ammonia concentrations at 25.0 deg C. Step constants of stability of studied complexes were calculated and their compositions were determined with account of nonideal character of aqueous-salt solutions with ammonia concentrations above 1.0 mol/dm 3 . Values of correction effects on salting out ammonia action for Bierrum function in solutions with 1.0-18 mol/dm 3 ammonia concentrations were found

  7. Ammonia/Hydrogen Mixtures in an SI-Engine

    DEFF Research Database (Denmark)

    Mørch, Christian Sandersen; Bjerre, Andreas; Gøttrup, Morten Piil

    2011-01-01

    In recent years there has been increasing focus on using metal ammine complexes for ammonia storage. In this paper a fuel system for ammonia fuelled internal combustion engines using metal ammine complexes as ammonia storage is analyzed. The use of ammonia/hydrogen mixtures as an SI-engine fuel...

  8. Ammonia as a Suitable Fuel for Fuel Cells

    International Nuclear Information System (INIS)

    Lan, Rong; Tao, Shanwen

    2014-01-01

    Ammonia, an important basic chemical, is produced at a scale of 150 million tons per year. Half of hydrogen produced in chemical industry is used for ammonia production. Ammonia containing 17.5 wt% hydrogen is an ideal carbon-free fuel for fuel cells. Compared to hydrogen, ammonia has many advantages. In this mini-review, the suitability of ammonia as fuel for fuel cells, the development of different types of fuel cells using ammonia as the fuel and the potential applications of ammonia fuel cells are briefly reviewed.

  9. Ammonia as a Suitable Fuel for Fuel Cells

    OpenAIRE

    Lan, Rong; Tao, Shanwen

    2014-01-01

    Ammonia, an important basic chemical, is produced at a scale of 150 million tons per year. Half of hydrogen produced in chemical industry is used for ammonia production. Ammonia containing 17.5 wt% hydrogen is an ideal carbon-free fuel for fuel cells. Compared to hydrogen, ammonia has many advantages. In this mini-review, the suitability of ammonia as fuel for fuel cells, the development of different types of fuel cells using ammonia as the fuel and the potential applications of ammonia fuel ...

  10. Ammonia as a suitable fuel for fuel cells

    Directory of Open Access Journals (Sweden)

    Rong eLan

    2014-08-01

    Full Text Available Ammonia, an important basic chemical, is produced at a scale of 150 million tons per year. Half of hydrogen produced in chemical industry is used for ammonia production. Ammonia containing 17.5wt% hydrogen is an ideal carbon-free fuel for fuel cells. Compared to hydrogen, ammonia has many advantages. In this mini-review, the suitability of ammonia as fuel for fuel cells, the development of different types of fuel cells using ammonia as the fuel and the potential applications of ammonia fuel cells are briefly reviewed.

  11. Aqueous Ammonia soaking of digested manure fibers

    DEFF Research Database (Denmark)

    Mirtsou-Xanthopoulou, Chrysoula; Jurado, Esperanza; Skiadas, Ioannis

    2012-01-01

    , their economical profitable operation relies on increasing the methane yield from manure, and especially of its solid fraction which is not so easily degradable. In the present study, Aqueous Ammonia Soaking was successfully applied on digested fibers separated from the effluent of a manure-fed, full......-scale anaerobic digester to enhance their methane productivity. Soaking in six different reagent concentrations in ammonia (5%, 10%, 15%, 20%, 25%, 32%) was applied for 3 days at 22°C. An overall methane yield increase from 85% to 110% was achieved compared to controls (digested manure fibers where AAS...... was not applied). The difference in reagent concentration at the range of 5-25% w/w in ammonia did not affect that much the overall methane yield resulting to an increase of 104-110% compared to the non AAS-treated fibers. Thus, an ammonia concentration as low as 5% is adequate for achieving the same increase...

  12. Ammonia Affects Astroglial Proliferation in Culture.

    Directory of Open Access Journals (Sweden)

    Guillermo Bodega

    Full Text Available Primary cultures of rat astroglial cells were exposed to 1, 3 and 5 mM NH4Cl for up to 10 days. Dose- and time-dependent reductions in cell numbers were seen, plus an increase in the proportion of cells in the S phase. The DNA content was reduced in the treated cells, and BrdU incorporation diminished. However, neither ammonia nor ammonia plus glutamine had any effect on DNA polymerase activity. iTRAQ analysis showed that exposure to ammonia induced a significant reduction in histone and heterochromatin protein 1 expression. A reduction in cell viability was also noted. The ammonia-induced reduction of proliferative activity in these cultured astroglial cells seems to be due to a delay in the completion of the S phase provoked by the inhibition of chromatin protein synthesis.

  13. Ammonia Affects Astroglial Proliferation in Culture

    Science.gov (United States)

    Bodega, Guillermo; Segura, Berta; Ciordia, Sergio; Mena, María del Carmen; López-Fernández, Luis Andrés; García, María Isabel; Trabado, Isabel; Suárez, Isabel

    2015-01-01

    Primary cultures of rat astroglial cells were exposed to 1, 3 and 5 mM NH4Cl for up to 10 days. Dose- and time-dependent reductions in cell numbers were seen, plus an increase in the proportion of cells in the S phase. The DNA content was reduced in the treated cells, and BrdU incorporation diminished. However, neither ammonia nor ammonia plus glutamine had any effect on DNA polymerase activity. iTRAQ analysis showed that exposure to ammonia induced a significant reduction in histone and heterochromatin protein 1 expression. A reduction in cell viability was also noted. The ammonia-induced reduction of proliferative activity in these cultured astroglial cells seems to be due to a delay in the completion of the S phase provoked by the inhibition of chromatin protein synthesis. PMID:26421615

  14. Insertion Mutation in HMG-CoA Lyase Increases the Production Yield of MPA through Agrobacterium tumefaciens-Mediated Transformation.

    Science.gov (United States)

    Dong, Yuguo; Zhang, Jian; Xu, Rui; Lv, Xinxin; Wang, Lihua; Sun, Aiyou; Wei, Dongzhi

    2016-11-28

    Mycophenolic acid (MPA) is an antibiotic produced by Penicillium brevicompactum . MPA has antifungal, antineoplastic, and immunosuppressive functions, among others. β-Hydroxy-β-methylglutaryl-CoA (HMG-CoA) lyase is a key enzyme in the bypass metabolic pathway. The inhibitory activity of HMG-CoA lyase increases the MPA biosynthetic flux by reducing the generation of by-products. In this study, we cloned the P. brevicompactum HMG-CoA lyase gene using the thermal asymmetric interlaced polymerase chain reaction and gene walking technology. Agrobacterium tumefaciens -mediated transformation (ATMT) was used to insert a mutated HMG-CoA lyase gene into P. brevicompactum . Successful insertion of the HMG-CoA lyase gene was confirmed by hygromycin screening, PCR, Southern blot analysis, and enzyme content assay. The maximum MPA production by transformants was 2.94 g/l. This was 71% higher than wild-type ATCC 16024. Our results demonstrate that ATMT may be an alternative practical genetic tool for directional transformation of P. brevicompactum .

  15. DIRECT AMMONIA-AIR FUEL CELL.

    Science.gov (United States)

    A new type of direct oxidation ammonia fuel cell was investigated. This cell is based on the use of a non-aqueous fused hydroxide electrolyte matrix...EMF’s of 0.5 to 0.6 volts. At practical levels of current density the direct ammonia fuel cell has an overall efficiency of about 60% compared to 30-35% for the indirect-type fuel cell . (Author)

  16. Nitrocarburizing in ammonia-hydrocarbon gas mixtures

    DEFF Research Database (Denmark)

    Pedersen, Hanne; Christiansen, Thomas; Somers, Marcel A. J.

    2011-01-01

    The present work investigates the possibility of nitrocarburising in ammonia-acetylene-hydrogen and ammonia-propene-hydrogen gas mixtures, where unsaturated hydrocarbon gas is the carbon source during nitrocarburising. Consequently, nitrocarburising is carried out in a reducing atmosphere...... microscopy and X-ray diffraction analysis. It is shown that the use of unsaturated hydrocarbon gas in nitrocarburising processes is a viable alternative to traditional nitrocarburising methods....

  17. Corrosion inhibitor for aqueous ammonia absorption system

    Science.gov (United States)

    Phillips, B.A.; Whitlow, E.P.

    1998-09-22

    A method is described for inhibiting corrosion and the formation of hydrogen and thus improving absorption in an ammonia/water absorption refrigeration, air conditioning or heat pump system by maintaining the hydroxyl ion concentration of the aqueous ammonia working fluid within a selected range under anaerobic conditions at temperatures up to 425 F. This hydroxyl ion concentration is maintained by introducing to the aqueous ammonia working fluid an inhibitor in an amount effective to produce a hydroxyl ion concentration corresponding to a normality of the inhibitor relative to the water content ranging from about 0.015 N to about 0.2 N at 25 C. Also, working fluids for inhibiting the corrosion of carbon steel and resulting hydrogen formation and improving absorption in an ammonia/water absorption system under anaerobic conditions at up to 425 F. The working fluids may be aqueous solutions of ammonia and a strong base or aqueous solutions of ammonia, a strong base, and a specified buffer. 5 figs.

  18. Effects of chronic ammonia exposure on ammonia metabolism and excretion in marine medaka Oryzias melastigma.

    Science.gov (United States)

    Gao, Na; Zhu, Limei; Guo, Zhiqiang; Yi, Meisheng; Zhang, Li

    2017-06-01

    Ammonia is highly toxic to aquatic organisms, but whether ammonia excretion or ammonia metabolism to less toxic compounds is the major strategy for detoxification in marine fish against chronic ammonia exposure is unclear to date. In this study, we investigated the metabolism and excretion of ammonia in marine medaka Oryzias melastigma during chronic ammonia exposure. The fish were exposed to 0, 0.1, 0.3, 0.6, and 1.1 mmol l -1  NH 4 Cl spiked seawater for 8 weeks. Exposure of 0.3-1.1 mmol l -1  NH 4 Cl had deleterious effects on the fish, including significant reductions in growth, feed intake, and total protein content. However, the fish could take strategies to detoxify ammonia. The tissue ammonia (T Amm ) in the 0.3-1.1 mmol l -1  NH 4 Cl treatments was significantly higher than those in the 0 and 0.1 mmol l -1  NH 4 Cl treatments after 2 weeks of exposure, but it recovered with prolonged exposure time, ultimately reaching the control level after 8 weeks. The amino acid catabolic rate decreased to reduce the gross ammonia production with the increasing ambient ammonia concentration. The concentrations of most metabolites remained constant in the 0-0.6 mmol l -1  NH 4 Cl treatments, whereas 5 amino acids and 3 energy metabolism-related metabolites decreased in the 1.1 mmol l -1  NH 4 Cl treatment. J Amm steadily increased in ambient ammonia from 0 to 0.6 mmol l -1 and slightly decreased when the ambient ammonia concentration increased to 1.1 mmol l -1 . Overall, marine medaka cope with sublethal ammonia environment by regulating the tissue T Amm via reducing the ammonia production and increasing ammonia excretion. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Sphingosine-1-Phosphate Lyase Deficient Cells as a Tool to Study Protein Lipid Interactions.

    Directory of Open Access Journals (Sweden)

    Mathias J Gerl

    Full Text Available Cell membranes contain hundreds to thousands of individual lipid species that are of structural importance but also specifically interact with proteins. Due to their highly controlled synthesis and role in signaling events sphingolipids are an intensely studied class of lipids. In order to investigate their metabolism and to study proteins interacting with sphingolipids, metabolic labeling based on photoactivatable sphingoid bases is the most straightforward approach. In order to monitor protein-lipid-crosslink products, sphingosine derivatives containing a reporter moiety, such as a radiolabel or a clickable group, are used. In normal cells, degradation of sphingoid bases via action of the checkpoint enzyme sphingosine-1-phosphate lyase occurs at position C2-C3 of the sphingoid base and channels the resulting hexadecenal into the glycerolipid biosynthesis pathway. In case the functionalized sphingosine looses the reporter moiety during its degradation, specificity towards sphingolipid labeling is maintained. In case degradation of a sphingosine derivative does not remove either the photoactivatable or reporter group from the resulting hexadecenal, specificity towards sphingolipid labeling can be achieved by blocking sphingosine-1-phosphate lyase activity and thus preventing sphingosine derivatives to be channeled into the sphingolipid-to-glycerolipid metabolic pathway. Here we report an approach using clustered, regularly interspaced, short palindromic repeats (CRISPR-associated nuclease Cas9 to create a sphingosine-1-phosphate lyase (SGPL1 HeLa knockout cell line to disrupt the sphingolipid-to-glycerolipid metabolic pathway. We found that the lipid and protein compositions as well as sphingolipid metabolism of SGPL1 knock-out HeLa cells only show little adaptations, which validates these cells as model systems to study transient protein-sphingolipid interactions.

  20. Mode of action of pectic enzymes. II. Further purification of exopolygalacturonate lyase and pectinesterase from Clostridium multifermentans.

    Science.gov (United States)

    Miller, L; Macmillan, J D

    1970-04-01

    Exopolygalacturonate lyase and pectinesterase from Clostridium multifermentans were purified 156-fold and 178-fold, respectively, by gel filtration chromatography on Sephadex G-200. The activities of both enzymes coincided in a single protein peak. Profiles of the two activities also coincided in diethylaminoethyl-cellulose chromatography and zonal centrifugation. These studies indicated that the esterase and the lyase were either complexed or similar molecular species. The former seems more probable because of the relatively high molecular weight. Both activities were most stable at pH 6.0. The esterase was inactivated rapidly at pH 5 or 7. Lyase preparations were freed of pectinesterase activity by heating for 30 min at 38 C and pH 7.0.

  1. [Sudden death of a patient with 3-hydroxy-3-methylglutaryl coenzyme A lyase deficiency].

    Science.gov (United States)

    Vilaseca Busca, M A; Ribes Rubio, A; Briones Godino, P; Cusi Sánchez, V; Baraíbar Castelló, R; Gairi Taull, J M

    1990-02-01

    A new case of neonatal 3-hydroxy-3-methylglutaric aciduria is described. 3-hydroxy-3-methylglutaryl CoA lyase activities in leukocytes demonstrated the patient's homozygosity and the heterozygous character of the parents and two other members of the family. Dietetic management with low fat high carbohydrate diet together with protein restriction and carnitine resulted in a good control of the metabolic acidosis, the hypoglycemia, and the physical and neurological development. Nevertheless, sudden death occurred at the age thirteen months without any previous apparent trouble and the necropsia showed neither signs of infection nor hepatic or cardiac derangement.

  2. One-step purification and characterization of alginate lyase from a clinical Pseudomonas aeruginosa with destructive activity on bacterial biofilm

    Directory of Open Access Journals (Sweden)

    Parinaz Ghadam

    2017-05-01

    Full Text Available Objective(s: Pseudomonas aeruginosais a Gram-negative and aerobic rod bacterium that displays mucoid and non-mucoid phenotype. Mucoid strains secrete alginate, which is the main agent of biofilms in chronic P. aeruginosa infections, show high resistance to antibiotics; consequently, the biological disruption of mucoid P. aeruginosa biofilms is an attractive area of study for researchers. Alginate lyase gene (algl is a member of alginate producing operon which by glycosidase activity produces primer for other enzymes in this cluster. Also this activity can destroy the extracellular alginate; therefore this enzyme participates in alginate production and destruction pathway. Alginate lyase causes detachment of a biofilm by reducing its adhesion to the surfaces, and increases phagocytosis and antibiotic susceptibility. In this study, alginate lyase was purified in just one step and its properties were investigated. Materials and Methods: The purification was done by affinity chromatography, analysed by SDS-PAGE, and its effect on P. aeruginosa biofilms was surveyed by micro titer plate assay and SEM. The substrate specificity of the enzyme was determined by PCR. Results: Alginate lyase from isolate 48 was purified in one step. It is more thermally resistant than alginate lyase from Pseudomonas aeruginosa PAO1 and poly M, poly G and poly MG alginate were the substrate of this enzyme. Moreover, it has an eradication effect on biofilms from P. aeruginosa 48 and PAO1. Conclusion: In this study an alginate lyase with many characteristics suitable in medicine such as thermal stability, effective on poly M alginate, and bacterial biofilm destructive was introduced and purified.

  3. Ammonia tolerant enriched methanogenic cultures as bioaugmentation inocula to alleviate ammonia inhibition in continuous anaerobic reactors

    DEFF Research Database (Denmark)

    Fotidis, Ioannis; Wang, Han; Angelidaki, Irini

    Ammonia is the most common inhibitor of anaerobic digestion (AD) process, resulting in suboptimal exploitation of the biogas potential of the feedstocks, causing significant economic losses to the biogas plants. Ammonia is mainly inhibiting the aceticlastic methanogens, while the hydrogenotrophic...... methanogens are more robust to ammonia toxicity effect. It has been shown that bioaugmentation of a pure strain of a hydrogenotrophic methanogen (i.e. Methanoculleus bourgensis) in an ammonia inhibited continuous anaerobic reactor can improve methane production more than 30%. Nevertheless, cultivation...... of a pure culture, to be used as bioaugmentation inoculum, poses technical difficulties due to the required sterile conditions and the special growing media. On the contrary acclimatized enrichment methanogenic cultures have lower requirements to sterility. In the present study, we used an enriched ammonia...

  4. A Case of Dilated Cardiomyopathy Associated with 3-Hydroxy-3-Methylglutaryl-Coenzyme A (HMG CoA Lyase Deficiency

    Directory of Open Access Journals (Sweden)

    Alexander A. C. Leung

    2009-01-01

    Full Text Available 3-hydroxy-3-methylglutaryl-coenzyme A (HMG CoA lyase deficiency is an inborn error of metabolism characterized by impairment of ketogenesis and leucine catabolism resulting in an organic acidopathy. In 1994, a case of dilated cardiomyopathy and fatal arrhythmia was reported in a 7-month-old infant. We report a case of dilated cardiomyopathy in association with HMG CoA lyase deficiency in a 23-year-old man with the acute presentation of heart failure. To our knowledge, this is the first case reported in an adult.

  5. Ammonia stripping of biologically treated liquid manure.

    Science.gov (United States)

    Alitalo, Anni; Kyrö, Aleksis; Aura, Erkki

    2012-01-01

    A prerequisite for efficient ammonia removal in air stripping is that the pH of the liquid to be stripped is sufficiently high. Swine manure pH is usually around 7. At pH 7 (at 20°C), only 0.4% of ammonium is in ammonia form, and it is necessary to raise the pH of swine slurry to achieve efficient ammonia removal. Because manure has a very high buffering capacity, large amounts of chemicals are needed to change the slurry pH. The present study showed that efficient air stripping of manure can be achieved with a small amount of chemicals and without strong bases like NaOH. Slurry was subjected to aerobic biological treatment to raise pH before stripping. This facilitated 8 to 32% ammonia removal without chemical treatment. The slurry was further subjected to repeated cycles of stripping with MgO and Ca(OH)(2) additions after the first and second strippings, respectively, to raise slurry pH in between the stripping cycles. After three consecutive stripping cycles, 59 to 86% of the original ammonium had been removed. It was shown that the reduction in buffer capacity of the slurry was due to ammonia and carbonate removal during the stripping cycles. Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.

  6. Study of Ammonia Emissions in a Ventilated Pig Pen

    DEFF Research Database (Denmark)

    Rong, Li

    Pig productions cause a wide emission of odors, such as ammonia (NH3), hydrogen sulfide (H2S), and methane (CH4). Ammonia is one of the most important emissions for evaluating the air quality either in animal buildings or atmospheric environment. In studies of ammonia emission from animal buildings...... reported in literature, little effort has been made to investigate the accuracy of current Henry’s law constant for modeling ammonia mass transfer process and study ammonia emissions in a full scale pig pen from fluid dynamics by CFD simulations. This will be the main objectives of this study. The ammonia...

  7. Effect of overexpression of fatty acid 9-hydroperoxide lyase in tomatoes (Lycopersicon esculentum Mill.).

    Science.gov (United States)

    Matsui, K; Fukutomi, S; Wilkinson, J; Hiatt, B; Knauf, V; Kajwara, T

    2001-11-01

    To modify the flavor properties of tomato fruits, cucumber fatty acid hydroperoxide lyase (HPL), which can act on 9-hydroperoxides of fatty acids to form volatile C9-aldehydes, was introduced to tomato plants. Through enzyme assay, high activity of the introduced HPL could be found in either the leaves or fruits of transgenic tomatoes; however, the composition of volatile short-chain aldehydes and alcohols in the transgenic tomato fruits was little modified. This was unexpected because tomato fruits have high lipoxygenase activity to form 9-hydroperoxides. When linoleic acid was added to a crude homogenate prepared from the transgenic tomato fruits, a high amount of C9-aldehyde was formed, but the amount of C6-aldehyde was almost equivalent to that in nontransgenic tomatoes. Through quantification of fatty acid hydroperoxides, it has been revealed that 13-hydroperoxides of fatty acids are preferably formed from endogenous substrate, whereas 9-hydroperoxides are formed from fatty acids added exogenously. From these observations, possible mechanisms to regulate metabolic flow of the lyase pathway are discussed.

  8. Enantioselective Synthesis of Various Cyanohydrins Using Covalently Immobilized Preparations of Hydroxynitrile Lyase from Prunus dulcis.

    Science.gov (United States)

    Alagöz, Dilek; Tükel, S Seyhan; Yildirim, Deniz

    2015-11-01

    The carrier-based and carrier-free (cross-linked enzyme aggregate) covalent immobilizations of Prunus dulcis hydroxynitrile lyase were investigated. The immobilized preparations were tested for enantioselective carbon-carbon bond formation activity in the biphasic medium. Of the tested preparations, only cross-linked enzyme aggregate of P. dulcis hydroxynitrile lyase (PdHNL-CLEA) achieved the synthesis of (R)-mandelonitrile with 93% yield and 99% enantiopurity. PdHNL-CLEA was also used in the synthesis of various (R)-cyanohydrins from corresponding aldehydes/ketones and hydrocyanic acid. When 4-methoxybenzaldehyde, 4-methyl benzaldehyde, and 4-hydroxybenzaldehyde were used as substrates, the yield-enantiomeric excess of corresponding (R)-cyanohydrins were obtained as 95-95, 85-79, and 2-25%, respectively, after 96 h at pH 4.0 and 5 °C. For acetophenone, 4-fluoroacetophenone, 4-chloroacetophenone, 4-bromoacetophenone, and 4-iodoacetophenone, the yield-enantiomeric excess of corresponding (R)-cyanohydrins were 1-99, 20-84, 11-95, 5-99, and 3-24%, respectively at the same conditions. The results demonstrate PdHNL-CLEA can be effectively used in the synthesis of (R)-mandelonitrile.

  9. Sugar-cane juice induces pectin lyase and polygalacturonase in Penicillium griseoroseum

    Directory of Open Access Journals (Sweden)

    Minussi Rosana Cristina

    1998-01-01

    Full Text Available The use of other inducers as substitutes for pectin was studied aiming to reduce the production costs of pectic enzymes. The effects of sugar-cane juice on the production of pectin lyase (PL and polygalacturonase (PG by Penicillium griseoroseum were investigated. The fungus was cultured in a mineral medium (pH 6.3 in a rotary shaker (150 rpm for 48 h at 25oC. Culture media were supplemented with yeast extract and sucrose or sugar-cane juice. Sugar-cane juice added singly to the medium promoted higher PL activity and mycelial dry weight when compared to pectin and the use of sugar-cane juice and yeast extract yielded levels of PG activity that were similar to those obtained with sucrose-yeast extract or pectin. The results indicated that, even at low concentrations, sugar-cane juice was capable of inducing pectin lyase and polygalacturonase with no cellulase activity in P. griseoroseum.

  10. Enzyme assay for identification of pectin and pectin derivatives, based on recombinant pectate lyase.

    Science.gov (United States)

    Hansen, K M; Truesen, A B; Søderberg, J R

    2001-01-01

    A simple method was developed for fast identification of pectin, based on a recombinant endopectate lyase cloned from Aspergillus niger. When pectin was demethylated and treated with pectate lyase, beta-elimination occurred, resulting in a double bond between C-4 and C-5 in the galacturonic acid residue of the released nonreducing end. The formation of double bonds produced an increase in light absorption, which was detected at 235 nm. The assay was tested on pectin of different origins (apple, orange, sugar beet, sunflower, celery, lemon), pectin derivatives (amidated pectin), and speciality types such as low molecular weight and low %DE (degree of esterification, percentage of galacturonic acid groups esterified with methanol) pectin. The highest response was given by pectate (pectin with %DEgums (carboxymethylcellulose, carrageenan, locust bean gum, tragacanth, gellan, tamarind, xanthan, amylogum, sodium alginate, or agar) gave any response. Members of IPPA (International Pectin Producers Association) have evaluated the validity of the assay in a ring test. All members of the Association were able to identify pectin from other gums in a blind test. The method can replace more laborious and ambiguous identification tests which exist today.

  11. Isolation and characterization of an Antarctic Flavobacterium strain with agarase and alginate lyase activities

    Directory of Open Access Journals (Sweden)

    Lavín Paris

    2016-09-01

    Full Text Available Several bacteria that are associated with macroalgae can use phycocolloids as a carbon source. Strain INACH002, isolated from decomposing Porphyra (Rhodophyta, in King George Island, Antarctica, was screened and characterized for the ability to produce agarase and alginate-lyase enzymatic activities. Our strain INACH002 was identified as a member of the genus Flavobacterium, closely related to Flavobacterium faecale, using 16S rRNA gene analysis. The INACH002 strain was characterized as psychrotrophic due to its optimal temperature (17ºC and maximum temperature (20°C of growth. Agarase and alginate-lyase displayed enzymatic activities within a range of 10°C to 50°C, with differences in the optimal temperature to hydrolyze agar (50°C, agarose (50°C and alginate (30°C during the first 30 min of activity. Strain Flavobacterium INACH002 is a promising Antarctic biotechnological resource; however, further research is required to illustrate the structural and functional bases of the enzymatic performance observed during the degradation of different substrates at different temperatures.

  12. Utilization of Glyphosate as Phosphate Source: Biochemistry and Genetics of Bacterial Carbon-Phosphorus Lyase

    Science.gov (United States)

    Zechel, David L.; Jochimsen, Bjarne

    2014-01-01

    SUMMARY After several decades of use of glyphosate, the active ingredient in weed killers such as Roundup, in fields, forests, and gardens, the biochemical pathway of transformation of glyphosate phosphorus to a useful phosphorus source for microorganisms has been disclosed. Glyphosate is a member of a large group of chemicals, phosphonic acids or phosphonates, which are characterized by a carbon-phosphorus bond. This is in contrast to the general phosphorus compounds utilized and metabolized by microorganisms. Here phosphorus is found as phosphoric acid or phosphate ion, phosphoric acid esters, or phosphoric acid anhydrides. The latter compounds contain phosphorus that is bound only to oxygen. Hydrolytic, oxidative, and radical-based mechanisms for carbon-phosphorus bond cleavage have been described. This review deals with the radical-based mechanism employed by the carbon-phosphorus lyase of the carbon-phosphorus lyase pathway, which involves reactions for activation of phosphonate, carbon-phosphorus bond cleavage, and further chemical transformation before a useful phosphate ion is generated in a series of seven or eight enzyme-catalyzed reactions. The phn genes, encoding the enzymes for this pathway, are widespread among bacterial species. The processes are described with emphasis on glyphosate as a substrate. Additionally, the catabolism of glyphosate is intimately connected with that of aminomethylphosphonate, which is also treated in this review. Results of physiological and genetic analyses are combined with those of bioinformatics analyses. PMID:24600043

  13. Abundance and genetic diversity of microbial polygalacturonase and pectate lyase in the sheep rumen ecosystem.

    Directory of Open Access Journals (Sweden)

    Peng Yuan

    Full Text Available Efficient degradation of pectin in the rumen is necessary for plant-based feed utilization. The objective of this study was to characterize the diversity, abundance, and functions of pectinases from microorganisms in the sheep rumen.A total of 103 unique fragments of polygalacturonase (PF00295 and pectate lyase (PF00544 and PF09492 genes were retrieved from microbial DNA in the rumen of a Small Tail Han sheep, and 66% of the sequences of these fragments had low identities (<65% with known sequences. Phylogenetic tree building separated the PF00295, PF00544, and PF09492 sequences into five, three, and three clades, respectively. Cellulolytic and noncellulolytic Butyrivibrio, Prevotella, and Fibrobacter species were the major sources of the pectinases. The two most abundant pectate lyase genes were cloned, and their protein products, expressed in Escherichia coli, were characterized. Both enzymes probably act extracellularly as their nucleotide sequences contained signal sequences, and they had optimal activities at the ruminal physiological temperature and complementary pH-dependent activity profiles.This study reveals the specificity, diversity, and abundance of pectinases in the rumen ecosystem and provides two additional ruminal pectinases for potential industrial use under physiological conditions.

  14. Expression and Bioinformatics Analysis of Pectate Lyase Gene from Bacillus subtilis521

    Science.gov (United States)

    Xiao, Jing; Lu, Fu-Ping; Li, Yu; Li, Jin-Ting

    In order to exploit new genetic resources, Pectate lyase(PEL) gene was amplified by PCR using the genome DNA from an alkaline Bacillus subtilis521. The PCR product was inserted into pET22b(+) vector. The recombinant plasmids were cloned in E.coli DH5α and then expressed in E.coli BL21. When cultured in the optimized medium, the positive clones E.coli BL21(pET22b(+)pel)showed intracellular pectate lyase activity of 90.0 U/mL. It was indicated that we had obtained the correct PEL gene. The pel has an open reading frame of 1263 nucleotides and codes for a product of 420 amino acids with a calculated molecular mass of 45.5 kD. Based on computer assisted analysis, a signal peptides and two conserved domains were revealed. The sequence analysis for PEL showed that it shares 26-82% homology with other strains in GenBank. In addition, the advanced structure of PEL were also predicted and analysed. This study will help to the experimental design of PEL fermentation and production purification and enzyme evolution.

  15. Molecular cloning, characterization and expression analysis of adenylosuccinate lyase gene in grass carp (Ctenopharyngodon idella).

    Science.gov (United States)

    Yuan, Tian; Gu, Ji-Rui; Gu, Wen-Bo; Wu, Jiang; Ge, Shao-Rong; Xu, Heng

    2011-03-01

    Adenylosuccinate lyase (ADSL) is a bifunctional enzyme acting in de novo purine synthesis and purine nucleotide recycling. In the present study, we have constructed a grass carp (Ctenopharyngodon idella) intestinal cDNA library that has over 2.3 × 10(5) primary clones. An expressed sequence tag (EST) of grass carp adenylosuccinate lyase (gcADSL) gene was screened from this library. Both 5'-RACE and 3'-RACE were carried out in order to obtain the complete cDNA sequence, which contains a 1,446 bp open reading frame encoding 482 amino acids about 54.552 kDa. The deduced amino acid sequence shares high homology with its vertebrate counterparts, which shares 94% similarity with zebrafish, 81% with African clawed frog as well as chicken, 77% with human and 76% with mouse. This gcADSL genomic sequence, consisted of 13 exons and 12 introns, is 8,557 bp in size. Real-time quantitative PCR analysis revealed that the highest expression level of gcADSL was detected in muscle and the lowest in gill. In western blotting analysis, His(6)-tagged gcADSL protein expressed in Escherichia coli could be recognized not only by an anti-His(6)-tag monoclonal antibody but also by an anti-human ADSL polyclonal antibody, indicating immunological crossreactivity occurs between grass carp and human ADSL protein. 1,082 bp 5'-flanking region sequence was cloned and analyzed.

  16. Structural Basis for Streptogramin B Resistance in Staphylococcus aureus by Virginiamycin B Lyase

    Energy Technology Data Exchange (ETDEWEB)

    Korczynska,M.; Mukhtar, T.; Wright, G.; Berghuis, A.

    2007-01-01

    The streptogramin combination therapy of quinupristin-dalfopristin (Synercid) is used to treat infections caused by bacterial pathogens, such as methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus faecium. However, the effectiveness of this therapy is being compromised because of an increased incidence of streptogramin resistance. One of the clinically observed mechanisms of resistance is enzymatic inactivation of the type B streptogramins, such as quinupristin, by a streptogramin B lyase, i.e., virginiamycin B lyase (Vgb). The enzyme catalyzes the linearization of the cyclic antibiotic via a cleavage that requires a divalent metal ion. Here, we present crystal structures of Vgb from S. aureus in its apoenzyme form and in complex with quinupristin and Mg{sup 2+} at 1.65- and 2.8-{angstrom} resolution, respectively. The fold of the enzyme is that of a seven-bladed {beta}-propeller, although the sequence reveals no similarity to other known members of this structural family. Quinupristin binds to a large depression on the surface of the enzyme, where it predominantly forms van der Waals interactions. Validated by site-directed mutagenesis studies, a reaction mechanism is proposed in which the initial abstraction of a proton is facilitated by a Mg{sup 2+}-linked conjugated system. Analysis of the Vgb-quinupristin structure and comparison with the complex between quinupristin and its natural target, the 50S ribosomal subunit, reveals features that can be exploited for developing streptogramins that are impervious to Vgb-mediated resistance.

  17. Expression, purification and crystallization of l-methionine γ-lyase 2 from Entamoeba histolytica

    Energy Technology Data Exchange (ETDEWEB)

    Sato, Dan [Department of Parasitology, Gunma University Graduate School of Medicine, 3-39-22 Showa-machi, Maebashi, Gunma 371-8511 (Japan); Yamagata, Wataru; Kamei, Kaeko [Graduate School of Science and Technology, Department of Applied Biology, Kyoto Institute of Technology, Sakyo-ku, Kyoto 606-8585 (Japan); Nozaki, Tomoyoshi [Department of Parasitology, Gunma University Graduate School of Medicine, 3-39-22 Showa-machi, Maebashi, Gunma 371-8511 (Japan); Harada, Shigeharu, E-mail: harada@kit.ac.jp [Graduate School of Science and Technology, Department of Applied Biology, Kyoto Institute of Technology, Sakyo-ku, Kyoto 606-8585 (Japan); Department of Parasitology, Gunma University Graduate School of Medicine, 3-39-22 Showa-machi, Maebashi, Gunma 371-8511 (Japan)

    2006-10-01

    l-Methionine γ-lyase 2 from E. histolytica, a key enzyme in sulfur-containing amino-acid degradation in this protozoan parasite, has been crystallized in a form suitable for X-ray structure analysis. l-Methionine γ-lyase (MGL) is considered to be an attractive target for rational drug development because the enzyme is absent in mammalian hosts. To enable structure-based design of drugs targeting MGL, one of the two MGL isoenzymes (EhMGL2) was crystallized in the orthorhombic space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 88.89, b = 102.68, c = 169.87 Å. The crystal diffracted to a resolution of 2.0 Å. The presence of a tetramer in the asymmetric unit (4 × 43.1 kDa) gives a Matthews coefficient of 2.2 Å{sup 3} Da{sup −1}. The structure was solved by the molecular-replacement method and structure refinement is now in progress.

  18. Effect of dietary protein restriction on renal ammonia metabolism

    Science.gov (United States)

    Lee, Hyun-Wook; Osis, Gunars; Handlogten, Mary E.; Guo, Hui; Verlander, Jill W.

    2015-01-01

    Dietary protein restriction has multiple benefits in kidney disease. Because protein intake is a major determinant of endogenous acid production, it is important that net acid excretion change in parallel during protein restriction. Ammonia is the primary component of net acid excretion, and inappropriate ammonia excretion can lead to negative nitrogen balance. Accordingly, we examined ammonia excretion in response to protein restriction and then we determined the molecular mechanism of the changes observed. Wild-type C57Bl/6 mice fed a 20% protein diet and then changed to 6% protein developed an 85% reduction in ammonia excretion within 2 days, which persisted during a 10-day study. The expression of multiple proteins involved in renal ammonia metabolism was altered, including the ammonia-generating enzymes phosphate-dependent glutaminase (PDG) and phosphoenolpyruvate carboxykinase (PEPCK) and the ammonia-metabolizing enzyme glutamine synthetase. Rhbg, an ammonia transporter, increased in expression in the inner stripe of outer medullary collecting duct intercalated cell (OMCDis-IC). However, collecting duct-specific Rhbg deletion did not alter the response to protein restriction. Rhcg deletion did not alter ammonia excretion in response to dietary protein restriction. These results indicate 1) dietary protein restriction decreases renal ammonia excretion through coordinated regulation of multiple components of ammonia metabolism; 2) increased Rhbg expression in the OMCDis-IC may indicate a biological role in addition to ammonia transport; and 3) Rhcg expression is not necessary to decrease ammonia excretion during dietary protein restriction. PMID:25925252

  19. Spatial and temporal variations in ammonia emissions

    DEFF Research Database (Denmark)

    Skjøth, Carsten Ambelas; Geels, Camilla; Berge, H

    2011-01-01

    Deriving a parameterisation of ammonia emissions for use in chemistry-transport models (CTMs) is a complex problem as the emission varies locally as a result of local climate and local agricultural management. In current CTMs such factors are generally not taken into account. This paper demonstra......Deriving a parameterisation of ammonia emissions for use in chemistry-transport models (CTMs) is a complex problem as the emission varies locally as a result of local climate and local agricultural management. In current CTMs such factors are generally not taken into account. This paper...... demonstrates how local climate and local management can be accounted for in CTMs by applying a modular approach for deriving data as input to a dynamic ammonia emission model for Europe. Default data are obtained from information in the RAINS system, and it is demonstrated how this dynamic emission model based...

  20. Enrichment of high ammonia tolerant methanogenic culture

    DEFF Research Database (Denmark)

    Fotidis, Ioannis; Karakashev, Dimitar Borisov; Proietti, Nicolas

    Ammonia is the major toxicant in full scale anaerobic digesters of animal wastes which are rich in proteins and/or urea, such as pig or poultry wastes. Ammonia inhibition decreases methane production rates, increases volatile fatty acids concentration and leads to economic losses for the biogas...... was derived from a full scale biogas reactor (Hashøj, Denmark), fed with 75% animal manure and 25% food industries organic waste. Basal anaerobic medium was used for the enrichment along with sodium acetate (1 g HAc L-1) as a carbon source. Fluorescence insitu hybridization (FISH) was used to determine...... microbial community composition. The outcome of the enrichment process was a mesophilic aceticlastic methanogenic enriched culture able to withstand high ammonia loads and utilize acetate and form methane stoichiometrically. FISH analysis showed that the methanogens of the enriched culture belonged...

  1. Ammonia emission time profiles based on manure transport data improve ammonia modelling across north western Europe

    Science.gov (United States)

    Hendriks, C.; Kranenburg, R.; Kuenen, J. J. P.; Van den Bril, B.; Verguts, V.; Schaap, M.

    2016-04-01

    Accurate modelling of mitigation measures for nitrogen deposition and secondary inorganic aerosol (SIA) episodes requires a detailed representation of emission patterns from agriculture. In this study the meteorological influence on the temporal variability of ammonia emissions from livestock housing and application of manure and fertilizer are included in the chemistry transport model LOTOS-EUROS. For manure application, manure transport data from Flanders (Belgium) were used as a proxy to derive the emission variability. Using improved ammonia emission variability strongly improves model performance for ammonia, mainly by a better representation of the spring maximum. The impact on model performance for SIA was negligible as explained by the limited, ammonia rich region in which the emission variability was updated. The contribution of Flemish agriculture to modelled annual mean ammonia and SIA concentrations in Flanders were quantified at respectively 7-8 and 1-2 μg/m3. A scenario study was performed to investigate the effects of reducing ammonia emissions from manure application during PM episodes by 75%, yielding a maximum reduction in modelled SIA levels of 1-3 μg/m3 during episodes. Year-to-year emission variability and a soil module to explicitly model the emission process from manure and fertilizer application are needed to further improve the modelling of the ammonia budget.

  2. Modelling of Ammonia Heat Pump Desuperheaters

    DEFF Research Database (Denmark)

    Christensen, Stefan Wuust; Elmegaard, Brian; Markussen, Wiebke Brix

    2015-01-01

    This paper presents a study of modelling desuperheating in ammonia heat pumps. Focus is on the temperature profile of the superheated refrigerant. Typically, the surface area of a heat exchanger is estimated using the Log Mean Temperature Difference (LMTD) method. The assumption of this method...... is that the specific heat is constant throughout the temperature glide of the refrigerant in the heat exchanger. However, considering ammonia as refrigerant, the LMTD method does not give accurate results due to significant variations of the specific heat. By comparing the actual temperature profiles from a one...

  3. Biochemistry of Ammonia Monoxygenase from Nitrosomonas

    Energy Technology Data Exchange (ETDEWEB)

    Alan Hooper

    2009-07-15

    Major results. 1. CytochromecM552, a protein in the electron transfer chain to ammonia monooxygenase. Purification, modeling of protein structure based on primary structure, characterization of 4 hemes by magnetic spectroscopy, potentiometry, ligand binding and turnover. Kim, H. J., ,Zatsman, et al. 2008). 2. Characterization of proteins which thought to be involved in the AMO reaction or to protect AMO from toxic nitrogenous intermediates such as NO. Nitrosocyanin is a protein present only in bacteria which catalyze the ammonia monoxygenase reaction (1). Cytochrome c P460 beta and cytochrome c’ beta.

  4. Electrochemical ammonia production on molybdenum nitride nanoclusters

    DEFF Research Database (Denmark)

    Howalt, Jakob Geelmuyden; Vegge, Tejs

    2013-01-01

    Theoretical investigations of electrochemical production of ammonia at ambient temperature and pressure on nitrogen covered molybdenum nanoparticles are presented. Density functional theory calculations are used in combination with the computational hydrogen electrode approach to calculate the free...... energy profile for electrochemical protonation of N2 and N adatoms on cuboctahedral Mo13 nanoparticles. Pathways for electrochemical ammonia production via direct protonation of N adatoms and N2 admolecules with an onset potential as low as -0.5 V and generally lower than -0.8 V on both a nitrogen...

  5. A 5-methylcytosine DNA glycosylase/lyase demethylates the retrotransposon Tos17 and promotes its transposition in rice

    KAUST Repository

    La, Honggui

    2011-09-06

    DNA 5-methylcytosine (5-meC) is an important epigenetic mark for transcriptional gene silencing in many eukaryotes. In Arabidopsis, 5-meC DNA glycosylase/lyases actively remove 5-meC to counter-act transcriptional gene silencing in a locus-specific manner, and have been suggested to maintain the expression of transposons. However, it is unclear whether plant DNA demethylases can promote the transposition of transposons. Here we report the functional characterization of the DNA glycosylase/lyase DNG701 in rice. DNG701 encodes a large (1,812 amino acid residues) DNA glycosylase domain protein. Recombinant DNG701 protein showed 5-meC DNA glycosylase and lyase activities in vitro. Knockout or knockdown of DNG701 in rice plants led to DNA hypermethylation and reduced expression of the retrotransposon Tos17. Tos17 showed less transposition in calli derived from dng701 knockout mutant seeds compared with that in wild-type calli. Overexpression of DNG701 in both rice calli and transgenic plants substantially reduced DNA methylation levels of Tos17 and enhanced its expression. The overexpression also led to more frequent transposition of Tos17 in calli. Our results demonstrate that rice DNG701 is a 5-meC DNA glycosylase/lyase responsible for the demethylation of Tos17 and this DNA demethylase plays a critical role in promoting Tos17 transposition in rice calli.

  6. Crystal Structure of α-1,4-Glucan Lyase, a Unique Glycoside Hydrolase Family Member with a Novel Catalytic Mechanism

    NARCIS (Netherlands)

    Rozeboom, Henriëtte J.; Yu, Shukun; Madrid, Susan; Kalk, Kor H.; Zhang, Ran; Dijkstra, Bauke W.

    2013-01-01

    α-1,4-Glucan lyase (EC 4.2.2.13) from the red seaweed Gracilariopsis lemaneiformis cleaves α-1,4-glucosidic linkages in glycogen, starch, and malto-oligosaccharides, yielding the keto-monosaccharide 1,5-anhydro-D-fructose. The enzyme belongs to glycoside hydrolase family 31 (GH31) but degrades

  7. Purification and characterization of a novel alginate lyase from the marine bacterium Cobetia sp. NAP1 isolated from brown algae.

    Science.gov (United States)

    Yagi, Hisashi; Fujise, Asako; Itabashi, Narumi; Ohshiro, Takashi

    2016-12-01

    The application of marine resources, instead of fossil fuels, for biomass production is important for building a sustainable society. Seaweed is valuable as a source of marine biomass for producing biofuels such as ethanol, and can be used in various fields. Alginate is an anionic polysaccharide that forms the main component of brown algae. Various alginate lyases (e.g. exo- and endo-types and oligoalginate lyase) are generally used to degrade alginate. We herein describe a novel alginate lyase, AlgC-PL7, which belongs to the polysaccharide lyase 7 family. AlgC-PL7 was isolated from the halophilic Gram-negative bacterium Cobetia sp. NAP1 collected from the brown algae Padina arborescens Holmes. The optimal temperature and pH for AlgC-PL7 activity were 45 °C and 8, respectively. Additionally, AlgC-PL7 was thermostable and salt-tolerant, exhibited broad substrate specificity, and degraded alginate into monosaccharides. Therefore, AlgC-PL7 is a promising enzyme for the production of biofuels.

  8. Structure of PhnP: a phosphodiesterase of the carbon-phosphorous lyase pathway for phosphonate degradation

    DEFF Research Database (Denmark)

    Podzelinska, Kateryna; He, Shu-Mei; Wathier, Matthew

    2009-01-01

    Carbon-phosphorus lyase is a multienzyme system encoded by the phn operon that enables bacteria to metabolize organophosphonates when the preferred nutrient, inorganic phosphate, is scarce. One of the enzymes encoded by this operon, PhnP, is predicted by sequence homology to be a metal-dependent ...

  9. Identification, expression, and characterization of a novel bacterial RGI Lyase enzyme for the production of bio-functional fibers

    DEFF Research Database (Denmark)

    da Silva, Ines Isabel Cardoso Rodrigues; Larsen, Dorte Møller; Meyer, Anne S.

    2011-01-01

    A gene encoding a putative rhamnogalacturonan I (RGI) Lyase (EC 4.2.2.-) from Bacillus licheniformis (DSM13) was selected after a homology search and phylogenetic analysis and optimized with respect to codon usage. The designed gene was transformed into Pichia pastoris and the enzyme was produced...

  10. Five phosphonate operon gene products as components of a multi-subunit complex of the carbon-phosphorus lyase pathway

    DEFF Research Database (Denmark)

    Jochimsen, Bjarne; Lolle, Signe; McSorley, Fern R.

    2011-01-01

    Organophosphonate utilization by Escherichia coli requires the 14 cistrons of the phnCDEFGHIJKLMNOP operon, of which the carbon-phosphorus lyase has been postulated to consist of the seven polypeptides specified by phnG to phnM. A 5,660-bp DNA fragment encompassing phnGHIJKLM is cloned, followed...

  11. Purification and Characterization of a Unique Pectin Lyase from Aspergillus giganteus Able to Release Unsaturated Monogalacturonate during Pectin Degradation

    Directory of Open Access Journals (Sweden)

    Danielle Biscaro Pedrolli

    2014-01-01

    Full Text Available A pectin lyase, named PLIII, was purified to homogeneity from the culture filtrate of Aspergillus giganteus grown in submerged culture containing orange peel waste as carbon source. PLIII was able to digest apple pectin and citrus pectins with different degrees of methyl esterification. Interestingly, the PLIII activity was stimulated in the presence of some divalent cations including Pb2+ and was not significantly affected by Hg2+. Like other pectin lyases, PLIII is stimulated by but is not dependent on Ca2+. The main soluble product released during the degradation of pectic substances promoted by the PLIII is compatible with an unsaturated monogalacturonate. PLIII is a unique enzyme able to release unsaturated monogalacturonate as the only soluble product during the degradation of pectic substances; therefore, PLIII was classified as an exo-pectin lyase. To our knowledge, this is the first characterization of an exo-pectin lyase. The PLIII described in this work is potentially useful for ethanol production from pectin-rich biomass, besides other common applications for alkaline pectinases like preparation of textile fibers, coffee and tea fermentation, vegetable oil extraction, and the treatment of pulp in papermaking.

  12. Purification and Characterization of a Unique Pectin Lyase from Aspergillus giganteus Able to Release Unsaturated Monogalacturonate during Pectin Degradation

    Science.gov (United States)

    Carmona, Eleonora Cano

    2014-01-01

    A pectin lyase, named PLIII, was purified to homogeneity from the culture filtrate of Aspergillus giganteus grown in submerged culture containing orange peel waste as carbon source. PLIII was able to digest apple pectin and citrus pectins with different degrees of methyl esterification. Interestingly, the PLIII activity was stimulated in the presence of some divalent cations including Pb2+ and was not significantly affected by Hg2+. Like other pectin lyases, PLIII is stimulated by but is not dependent on Ca2+. The main soluble product released during the degradation of pectic substances promoted by the PLIII is compatible with an unsaturated monogalacturonate. PLIII is a unique enzyme able to release unsaturated monogalacturonate as the only soluble product during the degradation of pectic substances; therefore, PLIII was classified as an exo-pectin lyase. To our knowledge, this is the first characterization of an exo-pectin lyase. The PLIII described in this work is potentially useful for ethanol production from pectin-rich biomass, besides other common applications for alkaline pectinases like preparation of textile fibers, coffee and tea fermentation, vegetable oil extraction, and the treatment of pulp in papermaking. PMID:25610636

  13. Mapping of the human adenylosuccinate lyase (ADSL) gene to chromosome 22q13.1-->q13.2.

    Science.gov (United States)

    Fon, E A; Demczuk, S; Delattre, O; Thomas, G; Rouleau, G A

    1993-01-01

    Adenylosuccinate lyase (ADSL) is an essential enzyme involved in de novo purine biosynthesis. A deficiency in ADSL in humans has been shown to predispose to a neurodevelopmental syndrome with autistic features. Using both a somatic cell hybrid mapping panel and fluorescence in situ hybridization, we have precisely localized the human ADSL gene to chromosome 22q13.1-->q13.2.

  14. The ketogenic diet is well tolerated and can be effective in patients with argininosuccinate lyase deficiency and refractory epilepsy

    NARCIS (Netherlands)

    Peuscher, Rosanne; Dijsselhof, Monique E.; Abeling, Nico G.; van Rijn, Margreet; van Spronsen, Francjan J.; Bosch, Annet M.

    2012-01-01

    Argininosuccinate lyase (ASL) deficiency (MIM 608310, McKusick 207900) is a rare disorder of the urea cycle, which leads to a deficiency of arginine and hyperammonemia. Epilepsy is a frequent complication of this disorder. A ketogenic diet (KD) can be a very effective therapy for refractory

  15. Activities of methionine-γ-lyase in the acidophilic archaeon “Ferroplasma acidarmanus” strain fer1

    Directory of Open Access Journals (Sweden)

    Khan MA

    2013-04-01

    Full Text Available M A Khan,1 Madeline M López-Muñoz,2 Charles W Kaspar,3 Kai F Hung1 1Department of Biological Sciences, Eastern Illinois University, Charleston, IL, USA; 2Department of Biology, Universidad de Puerto Rico, Mayaguez, Puerto Rico; 3Bacteriology Department, University of Wisconsin, Madison, WI, USA Abstract: Biogeochemical processes on exposed pyrite ores result in extremely high levels of sulfuric acid at these locations. Acidophiles that thrive in these conditions must overcome significant challenges, including an environment with proton concentrations at pH 3 or below. The role of sulfur metabolism in the archaeon “Ferroplasma acidarmanus” strain fer1's ability to thrive in this environment was investigated due to its growth-dependent production of methanethiol, a volatile organic sulfur compound. Two putative sequences for methionine-γ-lyase (EC 4.4.1.11, an enzyme known to carry out α, γ-elimination on L-methionine to produce methanethiol, were identified in fer1. Bioinformatic analyses identified a conserved pyridoxal-5'-phosphate (PLP binding domain and a partially conserved catalytic domain in both putative sequences. Detection of PLP-dependent and L-methionine-dependent production of α-keto compounds and thiol groups in fer1 confirmed the presence of methionine-γ-lyase activity. Further, fer1 lysate was capable of processing related substrates, including D-methionine, L-cysteine, L-cystathionine, and L/D-homocysteine. When the two putative fer1 methionine-γ-lyase gene-coded proteins were expressed in Escherichia coli cells, one sequence demonstrated an ability to carry out α, γ-elimination activity, while the other exhibited γ-replacement activity. These fer1 methionine-γ-lyases also exhibited optimum pH, substrate specificity, and catalytic preferences that are different from methionine-γ-lyases from other organisms. These differences are discussed in the context of molecular phylogeny constructed using a maximum

  16. An Intelligent Ammonia Sensor for Livestock Breeding Monitoring

    OpenAIRE

    Wang, Yang; Fu, Zetian; Zhang, Lingxian; Li, Xinxing; Xu, Dan; Zeng, Lihua; Ma, Juncheng; Peng, Fa

    2013-01-01

    International audience; Ammonia concentration is the major parameter to evaluate livestock breeding farms atmosphere quality and it also is regarded as the key indicator to describe the production of livestock breeding farms. Based on the oxidation characteristics of ammonia, this paper presented a new intelligent detecting instrument, the intelligent ammonia sensor, for the measurement of ammonia concentration, which used the microcontroller STM8L152 as the key control module. However, the T...

  17. Ammonia production, excretion, toxicity, and defense in fish: A Review

    Directory of Open Access Journals (Sweden)

    Alex Y K Ip

    2010-10-01

    Full Text Available Many fishes are ammonotelic but some species can detoxify ammonia to glutamine or urea. Certain fish species can accumulate high levels of ammonia in the brain or defense against ammonia toxicity by enhancing the effectiveness of ammonia excretion through active NH4+ transport, manipulation of ambient pH, or reduction in ammonia permeability through the branchial and cutaneous epithelia. Recent reports on ammonia toxicity in mammalian brain reveal the importance of permeation of ammonia through the blood-brain barrier and passages of ammonia and water through transporters in the plasmalemma of brain cells. Additionally, brain ammonia toxicity could be related to the passage of glutamine through the mitochondrial membranes into the mitochondrial matrix. On the other hand, recent reports on ammonia excretion in fish confirm the involvement of Rhesus glycoproteins in the branchial and cutaneous epithelia. Therefore, this review focuses on both the earlier literature and the up-to-date information on the problems and mechanisms concerning the permeation of ammonia, as NH3, NH4+ or proton-neutral nitrogenous compounds, across mitochondrial membranes, the blood-brain barrier, the plasmalemma of neurons, and the branchial and cutaneous epithelia of fish. It also addresses how certain fishes with high ammonia tolerance defend against ammonia toxicity through the regulation of the permeation of ammonia and related nitrogenous compounds through various types of membranes. It is hoped that this review would revive the interests in investigations on the passage of ammonia through the mitochondrial membranes and the blood-brain barrier of ammonotelic fishes and fishes with high brain ammonia-tolerance, respectively.

  18. 46 CFR 151.50-32 - Ammonia, anhydrous.

    Science.gov (United States)

    2010-10-01

    ... 46 Shipping 5 2010-10-01 2010-10-01 false Ammonia, anhydrous. 151.50-32 Section 151.50-32 Shipping... BULK LIQUID HAZARDOUS MATERIAL CARGOES Special Requirements § 151.50-32 Ammonia, anhydrous. (a) The anhydrous ammonia tanks may be installed in the bulk liquid cargo tanks provided the liquid surrounding the...

  19. Nitrite as a stimulus for ammonia-starved Nitrosomonas europaea

    NARCIS (Netherlands)

    Laanbroek, H.J.; Bär-Gilissen, M.J.; Hoogveld, H.L.

    2002-01-01

    Ammonia-starved cells of Nitrosomonas europaea are able to preserve a high level of ammonia-oxidizing activity in the absence of ammonium. However, when the nitrite-oxidizing cells that form part of the natural nitrifying community do not keep pace with the ammonia-oxidizing cells, nitrite

  20. Active site proton delivery and the lyase activity of human CYP17A1

    Energy Technology Data Exchange (ETDEWEB)

    Khatri, Yogan; Gregory, Michael C.; Grinkova, Yelena V.; Denisov, Ilia G.; Sligar, Stephen G., E-mail: s-sligar@illinois.edu

    2014-01-03

    Highlights: •The disruption of PREG/PROG hydroxylation activity by T306A showed the participation of Cpd I. •T306A supports the involvement of a nucleophilic peroxo-anion during lyase activity. •The presence of cytochrome b{sub 5} augments C–C lyase activity. •Δ5-Steroids are preferred substrates for CYP17 catalysis. -- Abstract: Cytochrome P450 CYP17A1 catalyzes a series of reactions that lie at the intersection of corticoid and androgen biosynthesis and thus occupies an essential role in steroid hormone metabolism. This multifunctional enzyme catalyzes the 17α-hydroxylation of Δ4- and Δ5-steroids progesterone and pregnenolone to form the corresponding 17α-hydroxy products through its hydroxylase activity, and a subsequent 17,20-carbon–carbon scission of pregnene-side chain produce the androgens androstenedione (AD) and dehydroepiandrosterone (DHEA). While the former hydroxylation reaction is believed to proceed through a conventional “Compound I” rebound mechanism, it has been suggested that the latter carbon cleavage is initiated by an iron-peroxy intermediate. We report on the role of Thr306 in CYP17 catalysis. Thr306 is a member of the conserved acid/alcohol pair thought to be essential for the efficient delivery of protons required for hydroperoxoanion heterolysis and formation of Compound I in the cytochromes P450. Wild type and T306A CYP17A1 self-assembled in Nanodiscs were used to quantitate turnover and coupling efficiencies of CYP17’s physiological Δ4- and Δ5-substrates. We observed that T306A co-incorporated in Nanodiscs with its redox partner cytochrome P450 oxidoreductase, coupled NADPH only by 0.9% and 0.7% compared to the wild type (97% and 22%) during the conversion of pregnenolone and progesterone, respectively, to the corresponding 17-OH products. Despite increased oxidation of pyridine nucleotide, hydroxylase activity was drastically diminished in the T306A mutant, suggesting a high degree of uncoupling in which reducing

  1. Structure-function analyses of a PL24 family ulvan lyase reveal key features and suggest its catalytic mechanism.

    Science.gov (United States)

    Ulaganathan, ThirumalaiSelvi; Helbert, William; Kopel, Moran; Banin, Ehud; Cygler, Miroslaw

    2018-01-30

    Ulvan is a major cell wall component of green algae of the genus Ulva and some marine bacteria encode enzymes that can degrade this polysaccharide. The first ulvan degrading lyases have been recently characterized and several putative ulvan lyases have been recombinantly expressed, confirmed as ulvan lyases and partially characterized. Two families of ulvan degrading lyases, PL24 and PL25, have recently been established. The PL24 lyase LOR_107 from the bacterial Alteromonadales sp. strain LOR degrades ulvan endolytically, cleaving the bond at the C4 of a glucuronic acid. However, the mechanism and LOR_107 structural features involved are unknown. We present here the crystal structure of LOR_107, representing the first PL24 family structure. We found that LOR_107 adopts a seven-bladed β-propeller fold with a deep canyon on one side of the protein. Comparative sequence analysis revealed a cluster of conserved residues within this canyon, and site-directed mutagenesis disclosed several residues essential for catalysis. We also found that LOR_107 uses the His/Tyr catalytic mechanism, common to several PL families. We captured a tetrasaccharide substrate in the structures of two inactive mutants, which indicated a two-step binding event, with the first substrate interaction near the top of the canyon coordinated by Arg-320, followed by sliding of the substrate into the canyon toward the active-site residues. Surprisingly, the LOR_107 structure was very similar to that of PL25 family PLSV_3936, despite only ~14% sequence identity between the two enzymes. On the basis of our structural and mutational analyses, we propose a catalytic mechanism for LOR_107 that differs from the typical His/Tyr mechanism. Published under license by The American Society for Biochemistry and Molecular Biology, Inc.

  2. Biochemical, Kinetic, and Spectroscopic Characterization of Ruegeria pomeroyi DddW--A Mononuclear Iron-Dependent DMSP Lyase.

    Directory of Open Access Journals (Sweden)

    Adam E Brummett

    Full Text Available The osmolyte dimethylsulfoniopropionate (DMSP is a key nutrient in marine environments and its catabolism by bacteria through enzymes known as DMSP lyases generates dimethylsulfide (DMS, a gas of importance in climate regulation, the sulfur cycle, and signaling to higher organisms. Despite the environmental significance of DMSP lyases, little is known about how they function at the mechanistic level. In this study we biochemically characterize DddW, a DMSP lyase from the model roseobacter Ruegeria pomeroyi DSS-3. DddW is a 16.9 kDa enzyme that contains a C-terminal cupin domain and liberates acrylate, a proton, and DMS from the DMSP substrate. Our studies show that as-purified DddW is a metalloenzyme, like the DddQ and DddP DMSP lyases, but contains an iron cofactor. The metal cofactor is essential for DddW DMSP lyase activity since addition of the metal chelator EDTA abolishes its enzymatic activity, as do substitution mutations of key metal-binding residues in the cupin motif (His81, His83, Glu87, and His121. Measurements of metal binding affinity and catalytic activity indicate that Fe(II is most likely the preferred catalytic metal ion with a nanomolar binding affinity. Stoichiometry studies suggest DddW requires one Fe(II per monomer. Electronic absorption and electron paramagnetic resonance (EPR studies show an interaction between NO and Fe(II-DddW, with NO binding to the EPR silent Fe(II site giving rise to an EPR active species (g = 4.29, 3.95, 2.00. The change in the rhombicity of the EPR signal is observed in the presence of DMSP, indicating that substrate binds to the iron site without displacing bound NO. This work provides insight into the mechanism of DMSP cleavage catalyzed by DddW.

  3. Ammonia emission time profiles based on manure transport data improve ammonia modelling across north western Europe

    NARCIS (Netherlands)

    Hendriks, C.; Kranenburg, R.; Kuenen, J.J.P.; Bril, B. van den; Verguts, V.; Schaap, M.

    2016-01-01

    Accurate modelling of mitigation measures for nitrogen deposition and secondary inorganic aerosol (SIA) episodes requires a detailed representation of emission patterns from agriculture. In this study the meteorological influence on the temporal variability of ammonia emissions from livestock

  4. Chemical Safety Alert: Hazards of Ammonia Releases at Ammonia Refrigeration Facilities

    Science.gov (United States)

    Anhydrous ammonia is used as a refrigerant in mechanical compression systems, often liquefied under pressure which increases exposure risk due to potential for rapid release into the air as a toxic gas.

  5. Planar optical waveguide sensor of ammonia

    Science.gov (United States)

    Sarkisov, Sergey S.; Curley, Michael J.; Boykin, Courtney; Diggs, Darnell E.; Grote, James G.; Hopkins, Frank K.

    2004-12-01

    We describe a novel sensor of ammonia based on a planar optical waveguide made of a thin film of polymer polyimide doped with indicator dye bromocresol purple. The film of dye-doped polyimide demonstrated reversible increase of absorption with a peak near 600 nm in response to presence of ammonia in ambient air. Coupling of input and output optic fibers with the waveguide was done by means of coupling prisms or coupling grooves. The latter configuration has the advantage of low cost, less sensitivity to temperature variation, and the possibility of coupling from both sides of the waveguide. Special experimental setup was built to test the sensor. It included test gas chamber with sealed optic fiber feed-throughs, gas filling line, laser source, photodetector, and signal processing hardware and software. The sensor was capable of detecting 100 ppm of ammonia in air within 8 seconds. Further increase of sensitivity can be achieved by adding more dye dopant to the polymer, increase of the length of the waveguide, and suppression of noise. Overexposure of the sensor to more than 5000 ppm of ammonia led to the saturation of the polymer film and, as a result, significant decrease of sensitivity and increase of the response time. The sensor can be used as low cost component of a distributed optical network of chemical sensors for monitoring presence of hazardous industrial pollutants in air.

  6. Effect of ammonia on Swiss albino mice

    Science.gov (United States)

    Hilado, C. J.; Casey, C. J.; Furst, A.

    1977-01-01

    Times to incapacitation and death and LC /50/ values were determined for Swiss albino male mice exposed to different concentrations of ammonia in a 4.2 liter hemispherical chamber. The LC/50/ for a 30 minute exposure was 21,430 ppm.

  7. The synthesis of ammonia from its elements

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 7; Issue 9. The Synthesis of Ammonia from its Elements. Fritz Haber. Classics Volume 7 Issue 9 September 2002 pp 86-94. Fulltext. Click here to view fulltext PDF. Permanent link: http://www.ias.ac.in/article/fulltext/reso/007/09/0086-0094 ...

  8. USDA-EPA Collaborative Ammonia Research

    Science.gov (United States)

    In 2014, a work group was formed between USDA and EPA to facilitate information exchange on ammonia emissions from agriculture, air quality impacts and emission mitigation options and to identify opportunities for collaboration. This document provides background on the work grou...

  9. Adsorption of Ammonia on Regenerable Carbon Sorbents

    Science.gov (United States)

    Wójtowicz, Marek A.; Cosgrove, Jesph E.; Serio, Michael A..; Wilburn, Monique

    2015-01-01

    Results are presented on the development of reversible sorbents for the combined carbon dioxide, moisture, and trace-contaminant (TC) removal for use in Extravehicular Activities (EVAs), and more specifically in the Primary Life Support System (PLSS). The currently available life support systems use separate units for carbon dioxide, trace contaminants, and moisture control, and the long-term objective is to replace the above three modules with a single one. Data on sorption and desorption of ammonia, which is a major TC of concern, are presented in this paper. The current TC-control technology involves the use of a packed bed of acid-impregnated granular charcoal, which is non-regenerable, and the carbon-based sorbent under development in this project can be regenerated by exposure to vacuum at room temperature. In this study, several carbon sorbents were fabricated and tested for ammonia sorption. Ammonia-sorption capacity was related to carbon pore structure characteristics, and the temperature of oxidative carbon-surface treatment was optimized for enhanced ammonia-sorption performance.

  10. Footprints on Ammonia Concentrations from Environmental Regulations

    DEFF Research Database (Denmark)

    Skjøth, Carsten Ambelas; Ellermann, Thomas; Hertel, Ole

    2008-01-01

    Releases of ammonia (NH3) to the atmosphere contribute significantly to the desposition of nitrogen to both terrestrial and aquatic ecosystems. This is the background for the national NH3 emission ceilings in Europe. However, in some countries the national legislation aims not only to meet theese...

  11. Radiation Chemistry in Ammonia-Water Ices

    Science.gov (United States)

    Loeffler, M. J.; Raut, U.; Baragiola, R. A.

    2010-01-01

    We studied the effects of 100 keV proton irradiation on films of ammonia-water mixtures between 20 and 120 K. Irradiation destroys ammonia, leading to the formation and trapping of H2, N2 NO, and N2O, the formation of cavities containing radiolytic gases, and ejection of molecules by sputtering. Using infrared spectroscopy, we show that at all temperatures the destruction of ammonia is substantial, but at higher temperatures (120 K), it is nearly complete (approximately 97% destroyed) after a fluence of 10(exp 16) ions per square centimeter. Using mass spectroscopy and microbalance gravimetry, we measure the sputtering yield of our sample and the main components of the sputtered flux. We find that the sputtering yield depends on fluence. At low temperatures, the yield is very low initially and increases quadratically with fluence, while at 120 K the yield is constant and higher initially. The increase in the sputtering yield with fluence is explained by the formation and trapping of the ammonia decay products, N2 and H2 which are seen to be ejected from the ice at all temperatures.

  12. Reducing ammonia volatilization from compound fertilizers ...

    African Journals Online (AJOL)

    Paul

    2012-09-13

    Sep 13, 2012 ... formulated fertilizer + 18.92 g zeolite (T5), 22.30 g 10:10:10 formulated fertilizer + 4.06 g zeolite (T6), and ... compound fertilizer could minimize ammonia loss. ..... North China Plan. Nutr. Cycl. Agroecosyst.63:187-195. Cottenie A (1980). Soil testing and plant testing as a basis of fertilizer recommendation.

  13. 1 Ammonia Concentrations in Different Aquaculture 2

    African Journals Online (AJOL)

    Administrator

    density, low water exchange rates, water source and the alkaline medium of the systems. Low unionized ammonia in earthen pond and natural pond was attributed to the presence of phytoplankton, high water exchanges, feeding system, low acidity and relatively low temperature. Remediating measures such as the use of ...

  14. Ammonia synthesis from first principles calculations

    DEFF Research Database (Denmark)

    Honkala, Johanna Karoliina; Hellman, Anders; Remediakis, Ioannis

    2005-01-01

    The rate of ammonia synthesis over a nanoparticle ruthenium catalyst can be calculated directly on the basis of a quantum chemical treatment of the problem using density functional theory. We compared the results to measured rates over a ruthenium catalyst supported on magnesium aluminum spinet...

  15. Orion Ammonia Boiler System Preflight Test Preparations

    Science.gov (United States)

    Levitt, Julia L.

    2017-01-01

    The Environmental Controls and Life Support Systems (ECLSS) branch at Kennedy Space Center (KSC) is currently undergoing preparations for ground testing of the Orion Multi-Purpose Crew Vehicle (MPCV) to prepare its subsystems for EM-1 (Exploration Mission-1). EM-1, Orions second unmanned flight, is a three-week long lunar mission during which the vehicle will complete a 6-day retrograde lunar orbit before returning to Earth. This paper focuses on the work done during the authors 16-week internship with the Mechanical Engineering Branch of KSCs Engineering Directorate. The authors project involved assisting with the preparations for testing the Orion MPCVs ammonia boiler system. The purpose of the ammonia boiler system is to keep the spacecraft sufficiently cool during the reentry portion of its mission, from service module (SM) separation to post-landing. This system is critical for keeping both the spacecraft (avionics and electronics) and crew alive during reentry, thus a successful test of the system is essential to the success of EM-1. XXXX The author was able to draft a detailed outline of the procedure for the ammonia system functional test. More work will need to be done on the vehicle power-up and power-down portions of the procedure, but the ammonia system testing portion of the procedure is thorough and includes vehicle test configurations, vehicle commands, and GSE. The author was able to compile a substantial list of questions regarding the ammonia system functional test with the help of her mentors. A significant number of these questions were answered in the teleconferences with Lockheed Martin.

  16. Ammonia and ammonium hydroxide sensors for ammonia/water absorption machines: Literature review and data compilation

    Science.gov (United States)

    Anheier, N. C., Jr.; McDonald, C. E.; Cuta, J. M.; Cuta, F. M.; Olsen, K. B.

    1995-05-01

    This report describes an evaluation of various sensing techniques for determining the ammonia concentration in the working fluid of ammonia/water absorption cycle systems. The purpose was to determine if any existing sensor technology or instrumentation could provide an accurate, reliable, and cost-effective continuous measure of ammonia concentration in water. The resulting information will be used for design optimization and cycle control in an ammonia-absorption heat pump. Pacific Northwest Laboratory (PNL) researchers evaluated each sensing technology against a set of general requirements characterizing the potential operating conditions within the absorption cycle. The criteria included the physical constraints for in situ operation, sensor characteristics, and sensor application. PNL performed an extensive literature search, which uncovered several promising sensing technologies that might be applicable to this problem. Sixty-two references were investigated, and 33 commercial vendors were identified as having ammonia sensors. The technologies for ammonia sensing are acoustic wave, refractive index, electrode, thermal, ion-selective field-effect transistor (ISFET), electrical conductivity, pH/colormetric, and optical absorption. Based on information acquired in the literature search, PNL recommends that follow-on activities focus on ISFET devices and a fiber optic evanescent sensor with a colormetric indicator. The ISFET and fiber optic evanescent sensor are inherently microminiature and capable of in situ measurements. Further, both techniques have been demonstrated selective to the ammonium ion (NH4(+)). The primary issue remaining is how to make the sensors sufficiently corrosion-resistant to be useful in practice.

  17. Rectal administration of 13N-ammonia in the study of ammonia metabolism

    International Nuclear Information System (INIS)

    Koen, Hirofumi

    1980-01-01

    13 N-ammonia produced by the cyclotron was instilled intrarectally in patients with liver diseases for the study of the turnover of rectally absorbed 13 N-ammonia. A positron camera connected to an on-line computer system was used for imaging of the liver and heart; 13 N-activity over the head was also recorded. Sequential changes of 13 N-activity in blood was measured, and chromatographic analysis of 13 N-labeled substances in blood was carried out using a Dowex 50W x 8 column. In the control, 13 N-ammonia was absorbed quickly into blood visualizing the liver shortly after administration, and hepatic uptake of 13 N-ammonia reached a plateau in 10 -- 15 min, whereas in patients with cirrhosis, the lung and heart were visualized in 5 min when the liver image was still faint. 13 N-activity over the head was apparently higher in the cirrhotic group. It was suggested that a large proportion of absorbed 13 N-ammonia bypassed liver cells and reached peripheral tissues. The heart/liver ratio of 13 N and 13 N over the head were closely correlated with various indices of portal hypertension. The relative proportion of 13 N-metabolites in blood was lower at 5 min and 15 min after administration in cirrhosis, suggesting a reduced capacity of the liver to remove and metabolize ammonia. (author)

  18. Ammonia and ammonium hydroxide sensors for ammonia/water absorption machines: Literature review and data compilation

    Energy Technology Data Exchange (ETDEWEB)

    Anheier, N.C. Jr.; McDonald, C.E.; Cuta, J.M.; Cuta, F.M.; Olsen, K.B.

    1995-05-01

    This report describes an evaluation of various sensing techniques for determining the ammonia concentration in the working fluid of ammonia/water absorption cycle systems. The purpose of this work was to determine if any existing sensor technology or instrumentation could provide an accurate, reliable, and cost-effective continuous measure of ammonia concentration in water. The resulting information will be used for design optimization and cycle control in an ammonia-absorption heat pump. PNL researchers evaluated each sensing technology against a set of general requirements characterizing the potential operating conditions within the absorption cycle. The criteria included the physical constraints for in situ operation, sensor characteristics, and sensor application. PNL performed an extensive literature search, which uncovered several promising sensing technologies that might be applicable to this problem. Sixty-two references were investigated, and 33 commercial vendors were identified as having ammonia sensors. The technologies for ammonia sensing are acoustic wave, refractive index, electrode, thermal, ion-selective field-effect transistor (ISFET), electrical conductivity, pH/colormetric, and optical absorption. Based on information acquired in the literature search, PNL recommends that follow-on activities focus on ISFET devices and a fiber optic evanescent sensor with a colormetric indicator. The ISFET and fiber optic evanescent sensor are inherently microminiature and capable of in situ measurements. Further, both techniques have been demonstrated selective to the ammonium ion (NH{sub 4}{sup +}). The primary issue remaining is how to make the sensors sufficiently corrosion-resistant to be useful in practice.

  19. ATP Citrate Lyase Regulates Myofiber Differentiation and Increases Regeneration by Altering Histone Acetylation

    Directory of Open Access Journals (Sweden)

    Suman Das

    2017-12-01

    Full Text Available ATP citrate lyase (ACL plays a key role in regulating mitochondrial function, as well as glucose and lipid metabolism in skeletal muscle. We report here that ACL silencing impairs myoblast and satellite cell (SC differentiation, and it is accompanied by a decrease in fast myosin heavy chain isoforms and MYOD. Conversely, overexpression of ACL enhances MYOD levels and promotes myogenesis. Myogenesis is dependent on transcriptional but also other mechanisms. We show that ACL regulates the net amount of acetyl groups available, leading to alterations in acetylation of H3(K9/14 and H3(K27 at the MYOD locus, thus increasing MYOD expression. ACL overexpression in murine skeletal muscle leads to improved regeneration after cardiotoxin-mediated damage. Thus, our findings suggest a mechanism for regulating SC differentiation and enhancing regeneration, which might be exploited for devising therapeutic approaches for treating skeletal muscle disease.

  20. Mechanism-based inactivator of isocitrate lyases 1 and 2 fromMycobacterium tuberculosis.

    Science.gov (United States)

    Pham, Truc V; Murkin, Andrew S; Moynihan, Margaret M; Harris, Lawrence; Tyler, Peter C; Shetty, Nishant; Sacchettini, James C; Huang, Hsiao-Ling; Meek, Thomas D

    2017-07-18

    Isocitrate lyase (ICL, types 1 and 2) is the first enzyme of the glyoxylate shunt, an essential pathway for Mycobacterium tuberculosis ( Mtb ) during the persistent phase of human TB infection. Here, we report 2-vinyl-d-isocitrate (2-VIC) as a mechanism-based inactivator of Mtb ICL1 and ICL2. The enzyme-catalyzed retro-aldol cleavage of 2-VIC unmasks a Michael substrate, 2-vinylglyoxylate, which then forms a slowly reversible, covalent adduct with the thiolate form of active-site Cys 191 2-VIC displayed kinetic properties consistent with covalent, mechanism-based inactivation of ICL1 and ICL2 with high efficiency (partition ratio, <1). Analysis of a complex of ICL1:2-VIC by electrospray ionization mass spectrometry and X-ray crystallography confirmed the formation of the predicted covalent S -homopyruvoyl adduct of the active-site Cys 191 .

  1. Enzyme discovery beyond homology: a unique hydroxynitrile lyase in the Bet v1 superfamily

    Science.gov (United States)

    Lanfranchi, Elisa; Pavkov-Keller, Tea; Koehler, Eva-Maria; Diepold, Matthias; Steiner, Kerstin; Darnhofer, Barbara; Hartler, Jürgen; van den Bergh, Tom; Joosten, Henk-Jan; Gruber-Khadjawi, Mandana; Thallinger, Gerhard G.; Birner-Gruenberger, Ruth; Gruber, Karl; Winkler, Margit; Glieder, Anton

    2017-05-01

    Homology and similarity based approaches are most widely used for the identification of new enzymes for biocatalysis. However, they are not suitable to find truly novel scaffolds with a desired function and this averts options and diversity. Hydroxynitrile lyases (HNLs) are an example of non-homologous isofunctional enzymes for the synthesis of chiral cyanohydrins. Due to their convergent evolution, finding new representatives is challenging. Here we show the discovery of unique HNL enzymes from the fern Davallia tyermannii by coalescence of transcriptomics, proteomics and enzymatic screening. It is the first protein with a Bet v1-like protein fold exhibiting HNL activity, and has a new catalytic center, as shown by protein crystallography. Biochemical properties of D. tyermannii HNLs open perspectives for the development of a complementary class of biocatalysts for the stereoselective synthesis of cyanohydrins. This work shows that systematic integration of -omics data facilitates discovery of enzymes with unpredictable sequences and helps to extend our knowledge about enzyme diversity.

  2. PMR6, a pectate lyase-like gene required for powdery mildew susceptibility in Arabidopsis.

    Science.gov (United States)

    Vogel, John P; Raab, Theodore K; Schiff, Celine; Somerville, Shauna C

    2002-09-01

    The plant genes required for the growth and reproduction of plant pathogens are largely unknown. In an effort to identify these genes, we isolated Arabidopsis mutants that do not support the normal growth of the powdery mildew pathogen Erysiphe cichoracearum. Here, we report on the cloning and characterization of one of these genes, PMR6. PMR6 encodes a pectate lyase-like protein with a novel C-terminal domain. Consistent with its predicted gene function, mutations in PMR6 alter the composition of the plant cell wall, as shown by Fourier transform infrared spectroscopy. pmr6-mediated resistance requires neither salicylic acid nor the ability to perceive jasmonic acid or ethylene, indicating that the resistance mechanism does not require the activation of well-described defense pathways. Thus, pmr6 resistance represents a novel form of disease resistance based on the loss of a gene required during a compatible interaction rather than the activation of known host defense pathways.

  3. Paraffin as oxygen vector modulates tyrosine phenol lyase production by Citrobacter freundii MTCC 2424.

    Science.gov (United States)

    Azmi, Wamik; Kumar, Ajay; Dev, Varun

    2013-06-01

    The efficiency of three oxygen-vectors liquid paraffin, silicone oil and n-dodecane in the production of tyrosine phenol lyase (TPL) by Citrobacter freundii MTCC 2424 was evaluated at 4% (v/v) concentration. The liquid paraffin as oxygenvectors was found to exhibit a stimulatory effect on TPL synthesis. The liquid paraffin at 6% (v/v) resulted in 34% increase in the TPL synthesis accompanied by a 13% increase in the production of cell mass at a 10 L scale. This improvement in TPL and cell mass production in the presence of liquid paraffin can be related to the fact that liquid paraffin was capable of maintaining dissolved O2 concentration above 28% throughout the course of the fermentation. Maintenance of the dissolved O2 concentration above 28% could be viewed in terms of an adequate oxygen supply to the rapidly dividing cells of the bacterium, which in turn resulted in enhanced synthesis of TPL and cell mass.

  4. Purine biosynthesis in Chinese Hamster cell mutants and human fibroblasts partially deficient in adenylosuccinate lyase

    International Nuclear Information System (INIS)

    Laikind, P.K.; Gruber, H.E.; Hoffer, M.; Jansen, I.; Miller, L.; Seegmiller, J.E.

    1986-01-01

    In a study of purine nucleotide metabolism of CHO-Ade I cells and fibroblasts from an adenylocsuccinate (AS) and SAICA riboside over-producing pattient the authors examined the effect of ASMP lyase deficiency on brachpoint enzyme activities and on de novo synthesis by comparison to the control cell lines, wild type parent cells (CHO-K1) and fibroblasts from age-matched normal individuals, respectively. The rate of newly-formed purine synthesis and excretion was measured by determining the amount of C 14-formate incorporatted into adenine, hypoxanthine, and guanasine moities. The nucleotide, nucleoside and base content of cells and media were analyzed by hhigh-pressure liquid chrromatography. Results are presented

  5. Gene deletion of cytosolic ATP: citrate lyase leads to altered organic acid production in Aspergillus niger

    DEFF Research Database (Denmark)

    Meijer, Susan Lisette; Nielsen, Michael Lynge; Olsson, Lisbeth

    2009-01-01

    With the availability of the genome sequence of the filamentous fungus Aspergillus niger, the use of targeted genetic modifications has become feasible. This, together with the fact that A. niger is well established industrially, makes this fungus an attractive micro-organism for creating a cell...... factory platform for production of chemicals. Using molecular biology techniques, this study focused on metabolic engineering of A. niger to manipulate its organic acid production in the direction of succinic acid. The gene target for complete gene deletion was cytosolic ATP: citrate lyase (acl), which...... the acl gene. Additionally, the total amount of organic acids produced in the deletion strain was significantly increased. Genome-scale stoichiometric metabolic model predictions can be used for identifying gene targets. Deletion of the acl led to increased succinic acid production by A. niger....

  6. The Skin Bacterium Propionibacterium acnes Employs Two Variants of Hyaluronate Lyase with Distinct Properties

    DEFF Research Database (Denmark)

    Nazipi, Seven; Stødkilde-Jørgensen, Kristian; Scavenius, Carsten

    2017-01-01

    Hyaluronic acid (HA) and other glycosaminoglycans are extracellular matrix components in the human epidermis and dermis. One of the most prevalent skin microorganisms, Propionibacterium acnes, possesses HA-degrading activity, possibly conferred by the enzyme hyaluronate lyase (HYL). In this study......, we identified the HYL of P. acnes and investigated the genotypic and phenotypic characteristics. Investigations include the generation of a P. acneshyl knockout mutant and HYL activity assays to determine the substrate range and formed products. We found that P. acnes employs two distinct variants...... of the observed differences between P. acnes phylotype IA and IB/II strains. Whereas type IA strains are primarily found on the skin surface and associated with acne vulgaris, type IB/II strains are more often associated with soft and deep tissue infections, which would require elaborate tissue invasion...

  7. The hydroperoxide lyase branch of the oxylipin pathway protects against photoinhibition of photosynthesis.

    Science.gov (United States)

    Savchenko, Tatyana; Yanykin, Denis; Khorobrykh, Andrew; Terentyev, Vasily; Klimov, Vyacheslav; Dehesh, Katayoon

    2017-06-01

    This study describes a new role for hydroperoxide lyase branch of oxylipin biosynthesis pathway in protecting photosynthetic apparatus under high light conditions. Lipid-derived signaling molecules, oxylipins, produced by a multi-branch pathway are central in regulation of a wide range of functions. The two most known branches, allene oxide synthase (AOS) and 13-hydroperoxide lyase (HPL) pathways, are best recognized as producers of defense compounds against biotic challenges. In the present work, we examine the role of these two oxylipin branches in plant tolerance to the abiotic stress, namely excessive light. Towards this goal, we have analyzed variable chlorophyll fluorescence parameters of intact leaves of Arabidopsis thaliana genotypes with altered oxylipin profile, followed by examining the impact of exogenous application of selected oxylipins on functional activity of photosynthetic apparatus in intact leaves and isolated thylakoid membranes. Our findings unequivocally bridge the function of oxylipins to photosynthetic processes. Specifically, HPL overexpressing lines display enhanced adaptability in response to high light treatment as evidenced by lower rate constant of photosystem 2 (PS2) photoinhibition and higher rate constant of PS2 recovery after photoinhibition. In addition, exogenous application of linolenic acid, 13-hydroperoxy linolenic acid, 12-oxophytodienoic acid, and methyl jasmonate individually, suppresses photochemical activity of PS2 in intact plants and isolated thylakoid membranes, while application of HPL-branch metabolites-does not. Collectively these data implicate function of HPL branch of oxylipin biosynthesis pathway in guarding PS2 under high light conditions, potentially exerted through tight regulation of free linolenic acid and 13-hydroperoxy linolenic acid levels, as well as competition with production of metabolites by AOS-branch of the oxylipin pathway.

  8. Lysine succinylation of Mycobacterium tuberculosis isocitrate lyase (ICL) fine-tunes the microbial resistance to antibiotics.

    Science.gov (United States)

    Zhou, Mingliang; Xie, Longxiang; Yang, Zhaozhen; Zhou, Jiahai; Xie, Jianping

    2017-04-01

    Lysine succinylation (Ksucc) is a newly identified protein posttranslational modification (PTM), which may play an important role in cellular physiology. However, the role of lysine succinylation in antibiotic resistance remains elusive. Isocitrate lyase (ICL) is crucial for broad-spectrum antibiotics tolerance in Mycobacterium tuberculosis (Mtb). We previously found that MtbICL (Rv0467) has at least three succinylated lysine residues, namely K189, K322, and K334.To explore the effect of succinylation on the activity of MtbICL, mutants' mimicry of the lysine succinylation were generated by site-directed mutagenesis. ICL-K189E mutant strain is more sensitive than the wild-type to rifampicin and streptomycin, but not isoniazid. For the in vitro activity of the purified isocitrate lyase, only K189E mutant showed significantly decreased activity. Crystal structure analysis showed that Lys189 Glu dramatically increased the pKa of Glu188 and decreased the pKa of Lys190, whereas had negligible effect on other residues within 5 Å as well as disruption of the electrostatic interaction between Lys189 and Glu182, which might prevent the closure of the active site loop and cause severe reduction of the enzyme activity. Considering the genetic, biochemical, and crystallographical evidences together, the succinylation of specific ICL residue can fine-tune the bacterial resistance to selected antibiotics. The decreased enzymatic activity resulting from the succinylation-changed electrostatic interaction might underlie this phenotype. This study provided the first insight into the link between lysine succinylation and antibiotic resistance.

  9. Cerebral ammonia uptake and accumulation during prolonged exercise in humans

    DEFF Research Database (Denmark)

    Nybo, Lars; Dalsgaard, Mads K.; Steensberg, Adam

    2005-01-01

    We evaluated whether peripheral ammonia production during prolonged exercise enhances the uptake and subsequent accumulation of ammonia within the brain. Two studies determined the cerebral uptake of ammonia (arterial and jugular venous blood sampling combined with Kety-Schmidt-determined cerebral...... blood flow; n = 5) and the ammonia concentration in the cerebrospinal fluid (CSF; n = 8) at rest and immediately following prolonged exercise either with or without glucose supplementation. There was a net balance of ammonia across the brain at rest and at 30 min of exercise, whereas 3 h of exercise...... exercise with glucose, and further to 16.1 ± 3.3 µM after the placebo trial (P

  10. Observations of atmospheric ammonia from TANSO-FTS/GOSAT

    Science.gov (United States)

    Someya, Yu; Imasu, Ryoichi; Saitoh, Naoko; Shiomi, Kei

    2017-04-01

    Atmospheric ammonia has large impacts on the nitrogen cycles or atmospheric environment such as nucleation of PM2.5 particles. It is reported that ammonia in the atmosphere has been increasing rapidly with the growth of population globally and this trend must continue in the future. Satellite observation is an effective approach to get to know the global perspectives of the gas. Atmospheric ammonia is observable using the thermal infrared (TIR) spectra, and IASI, TES and CrIS had been revealed those distributions. GOSAT also has TIR band including the ammonia absorption bands. GOSAT has the shorter revisit cycle than that of the other hyper-spectral TIR sounders mentioned above, therefore, the shorter time-scale events can be represented. In addition to the importance of the impacts of ammonia itself, the concentration ratio between ammonia and the other trace gases such as CO which is one of the main targets of the GOSAT-2 project is useful as the indicator of their emission sources. In this study, we introduce an algorithm to retrieve the column amount of atmospheric ammonia based on non-linear optimal estimation (Rogers, 2000) from GOSAT spectra in the ammonia absorption band between 960 - 970 cm-1. Temperature and water vapor profiles are estimated in advance of the ammonia retrieval. The preliminary results showed significant high concentrations of ammonia in the Northern India and the Eastern China as pointed out in the previous researches. We will discuss the global distribution of ammonia in the presentation.

  11. Contents and implications of ammonia human and canine bile.

    Science.gov (United States)

    Alderete, J S; Gaines, E L; Hudson, N L

    1978-08-01

    The quantification and physiological significance of ammonia in human and canine bile has not been clearly documented or analyzed previously. Bile from the gallbladder, common hepatic duct, and from T-tubes obtained from patients undergoing cholecystectomy and choledochotomy. Similar samples were obtained in 6 dogs by cannulation of their hepatic duct and various blood vessels. Simultaneous blood and bile samples were obtained at 15-min intervals before during, and after an exogenous load of ammonia, given intravenously. Ammonia levels were measured in all samples. Ammonia was measurable by conventional methods in human and canine bile. In both species the gallbladder bile contained at least twice as much ammonia as bile from the common hepatic duct. After an exogenous ammonia load, the levels of ammonia in canine bile increased in similar patterns, but in lesser magnitude than the ammonia levels measured at several sites in the circulation. The findings document that the bile contains ammonia in different quantities and suggest that bile could be an additional pathway for the removal of ammonia by the liver, in addition to the urea cycle. The loss of the biliary pathway for ammonia excretion could contribute to the pathophysiology of the signs and symptoms attributed to hyperammoniemia occurring in human beings and dogs with decreased hepatic function.

  12. Pretreatment of Biomass by Aqueous Ammonia for Bioethanol Production

    Science.gov (United States)

    Kim, Tae Hyun; Gupta, Rajesh; Lee, Y. Y.

    The methods of pretreatment of lignocellulosic biomass using aqueous ammonia are described. The main effect of ammonia treatment of biomass is delignification without significantly affecting the carbohydrate contents. It is a very effective pretreatment method especially for substrates that have low lignin contents such as agricultural residues and herbaceous feedstock. The ammonia-based pretreatment is well suited for simultaneous saccharification and co-fermentation (SSCF) because the treated biomass retains cellulose as well as hemicellulose. It has been demonstrated that overall ethanol yield above 75% of the theoretical maximum on the basis of total carbohydrate is achievable from corn stover pretreated with aqueous ammonia by way of SSCF. There are two different types of pretreatment methods based on aqueous ammonia: (1) high severity, low contact time process (ammonia recycle percolation; ARP), (2) low severity, high treatment time process (soaking in aqueous ammonia; SAA). Both of these methods are described and discussed for their features and effectiveness.

  13. Submersible microbial desalination cell for simultaneous ammonia recovery and electricity production from anaerobic reactors containing high levels of ammonia

    DEFF Research Database (Denmark)

    Zhang, Yifeng; Angelidaki, Irini

    2015-01-01

    High ammonia concentration in anaerobic reactors can seriously inhibit the anaerobic digestion process. In this study, a submersible microbial desalination cell (SMDC) was developed as an innovative method to lower the ammonia level in a continuous stirred tank reactor (CSTR) by in situ ammonia...... and free NH3 diffusion were identified as the mechanisms responsible for the ammonia transportation. With an increase in initial ammonia concentration and a decrease in external resistance, the SMDC performance was enhanced. In addition, the coexistence of other cations in CSTR or cathode had no negative...

  14. Ammonia in London: is it increasing and what is the relevance of urban ammonia for air quality impacts?

    Science.gov (United States)

    Braban, Christine; Tang, Sim; Poskitt, Janet; Van Dijk, Netty; Leeson, Sarah; Dragosits, Ulli; Hutchings, Torben; Twigg, Marsailidh; Di Marco, Chiara; Langford, Ben; Tremper, Anja; Nemitz, Eiko; Sutton, Mark

    2017-04-01

    Emissions of ammonia affect both rural and urban air quality primarily via reaction of ammonia in the atmosphere forming secondary ammonium salts in particulate matter (PM). Urban ammonia emissions come from a variety of sources including biological decomposition, human waste, industrial processes and combustion engines. In the UK, the only long-term urban ammonia measurement is a UK National Ammonia Monitoring Network site at London Cromwell Road, recording monthly average concentrations. Short term measurements have also been made in the past decade at Marylebone Road, North Kensington and on the BT Tower. Cromwell Road is a kerbside site operational since 1999. The Cromwell Road data indicates that ammonia concentrations may be increasing since 2010-2012 after a long period of decreasing. Data from the National Atmospheric Emissions Inventory indicates ammonia emissions from diesel fleet exhausts increasing over this time period but an overall net decrease in ammonia emissions. With changes in engine and exhaust technology to minimise pollutant emissions and the importance of ammonia as a precursor gas for secondary PM, there is a challenge to understand urban ammonia concentrations and subsequent impacts on urban air quality. In this paper the long term measurements are assessed in conjunction with the short-term measurements.The challenges to assess the relative importance of local versus long range ammonia emission are discussed.

  15. Innovative bioelectrochemical-anaerobic-digestion integrated system for ammonia recovery and bioenergy production from ammonia-rich residues

    DEFF Research Database (Denmark)

    Zhang, Yifeng; Angelidaki, Irini

    2015-01-01

    Ammonia (NH4+/NH3) inhibition during anaerobic digestion process is one of the most frequent problems existing in biogas plants, resulting in unstable process and reduced biogas production. In this study, we developed a novel hybrid system, consisted of a submersed microbial resource recovery cell...... (SMRC) and a continuous stirred tank reactor (CSTR), to prevent ammonia toxicity during anaerobic digestion by in-situ ammonia recovery and electricity production (Figure 1). In batch experiment, the ammonia concentration in the CSTR decreased from 6 to 0.7 g-N/L with an average recovery rate of 0.18 g...... of ammonia recovery on the microbial community composition in the integrated system. Results clearly indicate the great potential of the SMRC-CSTR-coupled system for efficient and cost-effective ammonia recovery, energy production and treatment of ammonia-rich residues....

  16. EFFECTIVE HYPERFINE-STRUCTURE FUNCTIONS OF AMMONIA

    International Nuclear Information System (INIS)

    Augustovičová, L.; Soldán, P.; Špirko, V.

    2016-01-01

    The hyperfine structure of the rotation-inversion ( v 2 = 0 + , 0 − , 1 + , 1 − ) states of the 14 NH 3 and 15 NH 3 ammonia isotopomers is rationalized in terms of effective (ro-inversional) hyperfine-structure (hfs) functions. These are determined by fitting to available experimental data using the Hougen’s effective hyperfine-structure Hamiltonian within the framework of the non-rigid inverter theory. Involving only a moderate number of mass independent fitting parameters, the fitted hfs functions provide a fairly close reproduction of a large majority of available experimental data, thus evidencing adequacy of these functions for reliable prediction. In future experiments, this may help us derive spectroscopic constants of observed inversion and rotation-inversion transitions deperturbed from hyperfine effects. The deperturbed band centers of ammonia come to the forefront of fundamental physics especially as the probes of a variable proton-to-electron mass ratio.

  17. Computational Search for Improved Ammonia Storage Materials

    DEFF Research Database (Denmark)

    Jensen, Peter Bjerre; Lysgaard, Steen; Vegge, Tejs

    , bromide or iodide, and mixtures thereof. In total the search space consists of thousands of combinations, which makes a GA ideal, to reduce the number of necessary calculations. We are screening for a one step release from either a hexa or octa ammine, and we have found promising candidates, which......Metal halide ammines, e.g. Mg(NH3)6Cl2 and Sr(NH3)8Cl2, can reversibly store ammonia, with high volumetric hydrogen storage capacities. The storage in the halide ammines is very safe, and the salts are therefore highly relevant as a carbon-free energy carrier in future transportation infrastructure....... In this project we are searching for improved mixed materials with optimal desorption temperatures and kinetics, optimally releasing all ammonia in one step. We apply Density Functional Theory, DFT, calculations on mixed compounds selected by a Genetic Algorithm (GA), relying on biological principles of natural...

  18. Experimental and numerical study of ammonia combustion

    OpenAIRE

    Duynslaegher, Catherine

    2011-01-01

    The problems of oil resources and CO2 emissions becoming increasingly alarming, the search for alternatives to fossil fuels is an important concern of our society. Even though hydrogen has been recognized as a promising fuel, implementing a global hydrogen-based economy is at present a non-feasible approach unless a suitable storage medium could be found. To bypass such difficulties, the use of ammonia in a modified spark ignition engine has been suggested. Since hydrogen must still be produc...

  19. Diverse microbial species survive high ammonia concentrations

    Science.gov (United States)

    Kelly, Laura C.; Cockell, Charles S.; Summers, Stephen

    2012-04-01

    Planetary protection regulations are in place to control the contamination of planets and moons with terrestrial micro-organisms in order to avoid jeopardizing future scientific investigations relating to the search for life. One environmental chemical factor of relevance in extraterrestrial environments, specifically in the moons of the outer solar system, is ammonia (NH3). Ammonia is known to be highly toxic to micro-organisms and may disrupt proton motive force, interfere with cellular redox reactions or cause an increase of cell pH. To test the survival potential of terrestrial micro-organisms exposed to such cold, ammonia-rich environments, and to judge whether current planetary protection regulations are sufficient, soil samples were exposed to concentrations of NH3 from 5 to 35% (v/v) at -80°C and room temperature for periods up to 11 months. Following exposure to 35% NH3, diverse spore-forming taxa survived, including representatives of the Firmicutes (Bacillus, Sporosarcina, Viridibacillus, Paenibacillus, Staphylococcus and Brevibacillus) and Actinobacteria (Streptomyces). Non-spore forming organisms also survived, including Proteobacteria (Pseudomonas) and Actinobacteria (Arthrobacter) that are known to have environmentally resistant resting states. Clostridium spp. were isolated from the exposed soil under anaerobic culture. High NH3 was shown to cause a reduction in viability of spores over time, but spore morphology was not visibly altered. In addition to its implications for planetary protection, these data show that a large number of bacteria, potentially including spore-forming pathogens, but also environmentally resistant non-spore-formers, can survive high ammonia concentrations.

  20. Ammonia volatilization from sows on grassland

    Science.gov (United States)

    Sommer, S. G.; Søgaard, H. T.; Møller, H. B.; Morsing, S.

    According to regulations, sows with piglets on organic farms must graze on pastures. Volatilization of ammonia (NH 3) from urine patches may represent a significant source of nitrogen (N) loss from these farms. Inputs of N are low on organic farms and losses may reduce crop production. This study examined spatial variations in NH 3 volatilization using a movable dynamic chamber, and the pH and total ammoniacal nitrogen (TAN) content in the topsoil of pastures with grazing sows was measured during five periods between June 1998 and May 1999. Gross NH 3 volatilization from the pastures was also measured with an atmospheric mass balance technique during seven periods from September 1997 until June 1999. The dynamic chamber study showed a high variation in NH 3 volatilization because of the distribution of urine; losses were between 0 and 2.8 g NH 3-N m -2 day -1. Volatilization was highest near the feeding area and the huts, where the sows tended to urinate. Ammonia volatilization rate was linearly related to the product of NH 3 concentration in the boundary layer and wind speed. The NH 3 in the boundary layer was in equilibrium with NH 3 in soil solution. Gross NH 3 volatilization was in the range 0.07-2.1 kg NH 3-N ha -1 day -1 from a pasture with 24 sows ha -1. Ammonia volatilization was related to the amount of feed given to the sows, incident solar radiation and air temperature during measuring periods, and also to temperature, incident solar radiation and rain 1-2 days before measurements. Annual ammonia loss was 4.8 kg NH 3-N sow -1.

  1. Nitrocarburising in ammonia-hydrocarbon gas mixtures

    DEFF Research Database (Denmark)

    Pedersen, Hanne; Christiansen, Thomas; Somers, Marcel A. J.

    2010-01-01

    The present work investigates the possibility of nitrocarburising in ammonia-acetylene-hydrogen and ammoniapropene- hydrogen gas mixtures, where unsaturated hydrocarbon gas is the carbon source during nitrocarburising. Consequently, nitrocarburising is carried out in a reducing atmosphere...... microscopy and X-ray diffraction analysis. It is shown that the use of unsaturated hydrocarbon gas in nitrocarburising processes is a viable alternative to traditional nitrocarburising methods....

  2. Ammonia Signaling in Yeast Colony Formation

    Czech Academy of Sciences Publication Activity Database

    Palková, Z.; Váchová, Libuše

    2003-01-01

    Roč. 225, - (2003), s. 229-272 ISSN 0074-7696 R&D Projects: GA ČR GA204/02/0650 Grant - others:GA of Charles University(CZ) 141/2001/B-BIO/PrF and EMBO YIP for ZP Institutional research plan: CEZ:MSM 113100003 Keywords : yeast colonies * ammonia * ammonium Subject RIV: EE - Microbiology, Virology Impact factor: 4.286, year: 2003

  3. Ammonia biofiltration and community analysis of ammonia-oxidizing bacteria in biofilters.

    Science.gov (United States)

    Jun, Yin; Wenfeng, Xu

    2009-09-01

    Biological removal of ammonia was investigated using compost and sludge as packing materials in laboratory-scale biofilters. The aim of this study is to characterize the composition of ammonia-oxidizing bacteria (AOB) in two biofilters designed to remove ammonia. Experimental tests and measurements included analysis of removal efficiency and metabolic products. The inlet concentration of ammonia applied was 20-100 mg m(-3). Removal efficiencies of BFC and BFS were in the range of 97-99% and 95-99%, respectively. Periodic analysis of the biofilter packing materials showed ammonia was removed from air stream by nitrification and by the improved absorption of NH(3) in the resultant acidity. Nitrate was the dominant product of NH(3) transformation. Changes in the composition of AOB were examined by using nested PCR, denaturing gradient gel electrophoresis (DGGE) and sequencing of DGGE bands. DGGE analysis of biofilter samples revealed that shifts in the community structure of AOB were observed in the experiment; however, the idle phase did not cause the structural shift of AOB. Phylogenetic analysis revealed the population of AOB showed Nitrosospira sp. remains the predominant population in BFC, while Nitrosomonas sp. is the predominant population in BFS.

  4. Peripheral Ammonia as a Mediator of Methamphetamine Neurotoxicity

    Science.gov (United States)

    Halpin, Laura E.; Yamamoto, Bryan K.

    2012-01-01

    Ammonia is metabolized by the liver and has established neurological effects. The current study examined the possibility that ammonia contributes to the neurotoxic effects of methamphetamine (METH). The results show that a binge dosing regimen of METH to the rat increased plasma and brain ammonia concentrations that were paralleled by evidence of hepatotoxicity. The role of peripheral ammonia in the neurotoxic effects of METH was further substantiated by the demonstration that the enhancement of peripheral ammonia excretion blocked the increases in brain and plasma ammonia and attenuated the long term depletions of dopamine and serotonin typically produced by METH. Conversely, the localized perfusion of ammonia in combination with METH, but not METH alone or ammonia alone, into the striatum recapitulated the neuronal damage produced by the systemic administration of METH. Furthermore, this damage produced by the local administration of ammonia and METH was blocked by the GYKI 52466, an AMPA receptor antagonist. These findings highlight the importance of ammonia derived from the periphery as a small molecule mediator of METH neurotoxicity and more broadly emphasize the importance of peripheral organ damage as a possible mechanism that mediates the neuropathology produced by drugs of abuse and other neuroactive molecules. PMID:22993432

  5. Ammonia recovery from anaerobically digested cattle manure by steam stripping.

    Science.gov (United States)

    Zeng, L; Mangan, C; Li, X

    2006-01-01

    Ammonia recovery from anaerobically digested cattle manure effluents through steam stripping was studied at a stripping tower temperature of 98-99 degrees C and a steam-water ratio approximately 56-72 g/L. The digested manure effluents were first treated by microfiltration and then the permeate was used as feed in steam stripping. The stripping performance was evaluated under different feed pH values, ammonia concentrations and temperatures. The increase of the initial feed pH does not significantly improve ammonia stripping efficiency due to the fact that the stripped effluent pH is increased during steam stripping. This suggests that steam stripping of anaerobically digested manure effluents for ammonia recovery may not need pre-raised pH. In contrast, the pH value of the synthetic ammonia wastewater containing NH4Cl dramatically decreases after steam stripping. Increasing the feed temperature slightly improves ammonia stripping efficiency, but reduces the concentration of the recovered ammonia in the condensate due to an increased condensate volume at a higher feed temperature. Therefore, the feed temperature should be controlled at an optimum point that can compromise the condensate ammonia concentration and the ammonia stripping efficiency. Experimental results indicate that recovery of ammonia from anaerobically digested cattle manure effluents as NH4OH is technically feasible.

  6. Analysis of the sulfur-regulated control of the cystathionine γ-lyase gene of Neurospora crassa

    Directory of Open Access Journals (Sweden)

    Reveal Brad S

    2012-07-01

    Full Text Available Abstract Background Cystathionine γ-lyase plays a key role in the transsulfuration pathway through its primary reaction of catalyzing the formation of cysteine from cystathionine. The Neurospora crassa cystathionine γ-lyase gene (cys-16+ is of particular interest in dissecting the regulation and dynamics of transsulfuration. The aim of this study was to determine the regulatory connection of cys-16+ to the Neurospora sulfur regulatory network. In addition, the cys-16+ promoter was characterized with the goal of developing a strongly expressed and regulatable gene expression tool. Findings The cystathionine γ-lyase cys-16+ gene was cloned and characterized. The gene, which contains no introns, encodes a protein of 417 amino acids with conserved pyridoxal 5’-phosphate binding site and substrate-cofactor binding pocket. Northern blot analysis using wild type cells showed that cys-16+ transcript levels increased under sulfur limiting (derepressing conditions and were present only at a low level under sulfur sufficient (repressing conditions. In contrast, cys-16+ transcript levels in a Δcys-3 regulatory mutant were present at a low level under either derepressing or repressing conditions. Gel mobility shift analysis demonstrated the presence of four CYS3 transcriptional activator binding sites on the cys-16+ promoter, which were close matches to the CYS3 consensus binding sequence. Conclusions In this work, we confirm the control of cystathionine γ-lyase gene expression by the CYS3 transcriptional activator through the loss of cys-16+ expression in a Δcys-3 mutant and through the in vitro binding of CYS3 to the cys-16+ promoter at four sites. The highly regulated cys-16+ promoter should be a useful tool for gene expression studies in Neurospora

  7. New Ulvan-Degrading Polysaccharide Lyase Family: Structure and Catalytic Mechanism Suggests Convergent Evolution of Active Site Architecture.

    Science.gov (United States)

    Ulaganathan, ThirumalaiSelvi; Boniecki, Michal T; Foran, Elizabeth; Buravenkov, Vitaliy; Mizrachi, Naama; Banin, Ehud; Helbert, William; Cygler, Miroslaw

    2017-05-19

    Ulvan is a complex sulfated polysaccharide biosynthesized by green seaweed and contains predominantly rhamnose, xylose, and uronic acid sugars. Ulvan-degrading enzymes have only recently been identified and added to the CAZy ( www.cazy.org ) database as family PL24, but neither their structure nor catalytic mechanism(s) are yet known. Several homologous, new ulvan lyases, have been discovered in Pseudoalteromonas sp. strain PLSV, Alteromonas LOR, and Nonlabens ulvanivorans, defining a new family PL25, with the lyase encoded by the gene PLSV_3936 being one of them. This enzyme cleaves the glycosidic bond between 3-sulfated rhamnose (R3S) and glucuronic acid (GlcA) or iduronic acid (IdoA) via a β-elimination mechanism. We report the crystal structure of PLSV_3936 and its complex with a tetrasaccharide substrate. PLSV_3936 folds into a seven-bladed β-propeller, with each blade consisting of four antiparallel β-strands. Sequence conservation analysis identified a highly conserved region lining at one end of a deep crevice on the protein surface. The putative active site was identified by mutagenesis and activity measurements. Crystal structure of the enzyme with a bound tetrasaccharide substrate confirmed the identity of base and acid residues and allowed determination of the catalytic mechanism and also the identification of residues neutralizing the uronic acid carboxylic group. The PLSV_3936 structure provides an example of a convergent evolution among polysaccharide lyases toward a common active site architecture embedded in distinct folds.

  8. Structural insights into RipC, a putative citrate lyase β subunit from a Yersinia pestis virulence operon

    International Nuclear Information System (INIS)

    Torres, Rodrigo; Chim, Nicholas; Sankaran, Banumathi; Pujol, Céline; Bliska, James B.; Goulding, Celia W.

    2011-01-01

    Comparison of the 2.45 Å resolution crystal structure of homotrimeric RipC, a putative citrate lyase β subunit from Y. pestis, with structural homologs reveals conserved RipC residues that are implicated in CoA binding. Yersinia pestis remains a threat, with outbreaks of plague occurring in rural areas and its emergence as a weapon of bioterrorism; thus, an improved understanding of its various pathogenicity pathways is warranted. The rip (required for intracellular proliferation) virulence operon is required for Y. pestis survival in interferon-γ-treated macrophages and has been implicated in lowering macrophage-produced nitric oxide levels. RipC, one of three gene products from the rip operon, is annotated as a citrate lyase β subunit. Furthermore, the Y. pestis genome lacks genes that encode citrate lyase α and γ subunits, suggesting a unique functional role of RipC in the Y. pestisrip-mediated survival pathway. Here, the 2.45 Å resolution crystal structure of RipC revealed a homotrimer in which each monomer consists of a (β/α) 8 TIM-barrel fold. Furthermore, the trimeric state was confirmed in solution by size-exclusion chromatography. Through sequence and structure comparisons with homologous proteins, it is proposed that RipC is a putative CoA- or CoA-derivative binding protein

  9. A Systems Chemical Biology Study of Malate Synthase and Isocitrate Lyase Inhibition in Mycobacterium tuberculosis During Active and NRP Growth

    Science.gov (United States)

    May, Elebeoba E.; Leitão, Andrei; Tropsha, Alexander; Oprea, Tudor I.

    2013-01-01

    The ability of Mycobacterium tuberculosis (Mtb) to survive in low oxygen environments enables the bacterium to persist in a latent state within host tissues. In vitro studies of Mtb growth have identified changes in isocitrate lyase (ICL) and malate synthase (MS) that enable bacterial persistent under low oxygen and other environmentally limiting conditions. Systems chemical biology (SCB) enables us to evaluate the effects of small molecule inhibitors not only on the reaction catalyzed by malate synthase and isocitrate lyase, but the effect on the complete tricarboxylic acid cycle (TCA) by taking into account complex network relationships within that system. To study the kinetic consequences of inhibition on persistent bacilli, we implement a systems-chemical biology (SCB) platform and perform a chemistry-centric analysis of key metabolic pathways believed to impact Mtb latency. We explore consequences of disrupting the function of malate synthase (MS) and isocitrate lyase (ICL) during aerobic and hypoxic non-replicating persistence (NRP) growth by using the SCB method to identify small molecules that inhibit the function of MS and ICL, and simulating the metabolic consequence of the disruption. Results indicate variations in target and non-target reaction steps, clear differences in the normal and low oxygen models, as well as dosage dependent response. Simulation results from singular and combined enzyme inhibition strategies suggest ICL may be the more effective target for chemotherapeutic treatment against Mtb growing in a microenvironment where oxygen is slowly depleted, which may favor persistence. PMID:24121675

  10. Antisense inhibition of a pectate lyase gene supports a role for pectin depolymerization in strawberry fruit softening.

    Science.gov (United States)

    Santiago-Doménech, Nieves; Jiménez-Bemúdez, Silvia; Matas, Antonio J; Rose, Jocelyn K C; Muñoz-Blanco, Juan; Mercado, José A; Quesada, Miguel A

    2008-01-01

    Cell wall disassembly in softening fruits is a complex process involving the cumulative action of many families of wall-modifying proteins on interconnected polysaccharide matrices. One strategy to elucidate the in vivo substrates of specific enzymes and their relative importance and contribution to wall modification is to suppress their expression in transgenic fruit. It has been reported previously that inhibiting the expression of pectate lyase genes by antisense technology in strawberry (Fragaria x ananassa Duch.) fruit resulted in prolonged fruit firmness. This suggested that pectin depolymerization might make a more important contribution to strawberry fruit softening than is often stated. In this present study, three independent transgenic lines were identified exhibiting a greater than 90% reduction in pectate lyase transcript abundance. Analyses of sequential cell wall extracts from the transgenic and control fruit collectively showed clear quantitative and qualitative differences in the extractability and molecular masses of populations of pectin polymers. Wall extracts from transgenic fruits showed a reduction in pectin solubility and decreased depolymerization of more tightly bound polyuronides. Additional patterns of differential extraction of other wall-associated pectin subclasses were apparent, particularly in the sodium carbonate- and chelator-soluble polymers. In addition, microscopic studies revealed that the typical ripening-associated loss of cell-cell adhesion was substantially reduced in the transgenic fruits. These results indicate that pectate lyase plays an important degradative role in the primary wall and middle lamella in ripening strawberry fruit, and should be included in synergistic models of cell wall disassembly.

  11. Lactic acid bacteria involved in cocoa beans fermentation from Ivory Coast: Species diversity and citrate lyase production.

    Science.gov (United States)

    Ouattara, Hadja D; Ouattara, Honoré G; Droux, Michel; Reverchon, Sylvie; Nasser, William; Niamke, Sébastien L

    2017-09-01

    Microbial fermentation is an indispensable process for high quality chocolate from cocoa bean raw material. lactic acid bacteria (LAB) are among the major microorganisms responsible for cocoa fermentation but their exact role remains to be elucidated. In this study, we analyzed the diversity of LAB in six cocoa producing regions of Ivory Coast. Ribosomal 16S gene sequence analysis showed that Lactobacillus plantarum and Leuconostoc mesenteroides are the dominant LAB species in these six regions. In addition, other species were identified as the minor microbial population, namely Lactobacillus curieae, Enterococcus faecium, Fructobacillus pseudoficulneus, Lactobacillus casei, Weissella paramesenteroides and Weissella cibaria. However, in each region, the LAB microbial population was composed of a restricted number of species (maximum 5 species), which varied between the different regions. LAB implication in the breakdown of citric acid was investigated as a fundamental property for a successful cocoa fermentation process. High citrate lyase producer strains were characterized by rapid citric acid consumption, as revealed by a 4-fold decrease in citric acid concentration in the growth medium within 12h, concomitant with an increase in acetic acid and lactic acid concentration. The production of citrate lyase was strongly dependent on environmental conditions, with optimum production at acidic pH (pHcocoa fermentation. This study reveals that one of the major roles of LAB in the cocoa fermentation process involves the breakdown of citric acid during the early stage of cocoa fermentation through the activity of citrate lyase. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Kinetic model of ammonia synthesis in the solar nebula

    Science.gov (United States)

    Norris, T. L.

    1980-03-01

    Kinetic model of ammonia formation by iron catalysis in the primordial solar nebula is developed. The maximum time to reach equilibrium concentration is determined for various temperatures between 1000 and 200 K on the basis of reaction rates derived from industrial data on iron catalysts for ammonia. Application of the method for calculating the equilibrium time to an arbitrary nebula cooling model which maximizes the time available for ammonia synthesis results in an upper limit of 3% of the equilibrium value to the proportion of nitrogen in the form of ammonia at the time of planetary accretion, with ammonia abundance decreasing with distance from the sun. It is concluded that kinetic rather than equilibrium considerations control the abundance of ammonia in the solar nebula, and implications of the dominance of nitrogen for the evolution of the atmospheres of the terrestrial and Jovian planets and the composition of comets are indicated.

  13. Converting Wind Energy to Ammonia at Lower Pressure

    International Nuclear Information System (INIS)

    Malmali, Mahdi; Reese, Michael; McCormick, Alon V.; Cussler, E. L.

    2017-01-01

    Renewable wind energy can be used to make ammonia. However, wind-generated ammonia costs about twice that made from a traditional fossil-fuel driven process. To reduce the production cost, we replace the conventional ammonia condensation with a selective absorber containing metal halides, e.g., calcium chloride, operating at near synthesis temperatures. With this reaction-absorption process, ammonia can be synthesized at 20 bar from air, water, and wind-generated electricity, with rates comparable to the conventional process running at 150–300 bar. In our reaction-absorption process, the rate of ammonia synthesis is now controlled not by the chemical reaction but largely by the pump used to recycle the unreacted gases. The results suggest an alternative route to distributed ammonia manufacture which can locally supply nitrogen fertilizer and also a method to capture stranded wind energy as a carbon-neutral liquid fuel.

  14. Ammonia in the environment: From ancient times to the present

    Energy Technology Data Exchange (ETDEWEB)

    Sutton, M.A.; Erisman, J.W.; Dentener, F.; Moller, D. [Centre for Ecology and Hydrology, Penicuik (United Kingdom). Edinburgh Research Station

    2008-12-15

    Recent research on atmospheric ammonia has made good progress in quantifying sources/sinks and environmental impacts. This paper reviews the achievements and places them in their historical context. It considers the role of ammonia in the development of agricultural science and air chemistry, showing how these arose out of foundations in 18th century chemistry and medieval alchemy, and then identifies the original environmental sources from which the ancients obtained ammonia. Ammonia is revealed as a compound of key human interest through the centuries, with a central role played by sal ammoniac in alchemy and the emergence of modern science. The review highlights how recent environmental research has emphasized volatilization sources of ammonia. Conversely, the historical records emphasize the role of high-temperature sources, including dung burning, coal burning, naturally burning coal seams and volcanoes. Present estimates of ammonia emissions from these sources are based on few measurements, which should be a future priority.

  15. Ammonia in the environment: from ancient times to the present.

    Science.gov (United States)

    Sutton, Mark A; Erisman, Jan Willem; Dentener, Frank; Möller, Detlev

    2008-12-01

    Recent research on atmospheric ammonia has made good progress in quantifying sources/sinks and environmental impacts. This paper reviews the achievements and places them in their historical context. It considers the role of ammonia in the development of agricultural science and air chemistry, showing how these arose out of foundations in 18th century chemistry and medieval alchemy, and then identifies the original environmental sources from which the ancients obtained ammonia. Ammonia is revealed as a compound of key human interest through the centuries, with a central role played by sal ammoniac in alchemy and the emergence of modern science. The review highlights how recent environmental research has emphasized volatilization sources of ammonia. Conversely, the historical records emphasize the role of high-temperature sources, including dung burning, coal burning, naturally burning coal seams and volcanoes. Present estimates of ammonia emissions from these sources are based on few measurements, which should be a future priority.

  16. Process model for ammonia volatilization from anaerobic swine lagoons incorporating varying wind speeds and biogas bubbling

    Science.gov (United States)

    Ammonia volatilization from treatment lagoons varies widely with the total ammonia concentration, pH, temperature, suspended solids, atmospheric ammonia concentration above the water surface, and wind speed. Ammonia emissions were estimated with a process-based mechanistic model integrating ammonia ...

  17. Calculation notes in support of ammonia releases from waste tank ventilation systems

    International Nuclear Information System (INIS)

    Wojdac, L.F.

    1996-01-01

    Ammonia is generated in waste tanks via the degradation of nitrogen compounds. The ammonia is released from the liquids by a mechanism which is dependent on temperature, pH, ionic strength and ammonia concentration. The release of ammonia to the environment occurs via diffusion of ammonia through a stagnant air mass and into the ventilation system

  18. 46 CFR 98.25-5 - How anhydrous ammonia may be carried.

    Science.gov (United States)

    2010-10-01

    ... 46 Shipping 4 2010-10-01 2010-10-01 false How anhydrous ammonia may be carried. 98.25-5 Section 98... Anhydrous Ammonia in Bulk § 98.25-5 How anhydrous ammonia may be carried. (a) Anhydrous ammonia shall be..., except as otherwise provided in paragraph (b) of this section. (b) When anhydrous ammonia is to be...

  19. Upwind impacts of ammonia from an intensive poultry unit

    International Nuclear Information System (INIS)

    Jones, L.; Nizam, M.S.; Reynolds, B.; Bareham, S.; Oxley, E.R.B.

    2013-01-01

    This study investigated potential ammonia impacts on a sand dune nature reserve 600 m upwind of an intensive poultry unit. Ammonia concentrations and total nitrogen deposition were measured over a calendar year. A series of ammonia and nitrogen exposure experiments using dune grassland species were conducted in controlled manipulations and in the field. Ammonia emissions from the intensive poultry unit were detected up to 2.8 km upwind, contributing to exceedance of critical levels of ammonia 800 m upwind and exceedance of critical loads of nitrogen 2.8 km upwind. Emissions contributed 30% of the total N load in parts of the upwind conservation site. In the nitrogen exposure experiments, plants showed elevated tissue nitrogen contents, and responded to ammonia concentrations and nitrogen deposition loads observed in the conservation site by increasing biomass. Estimated long-term impacts suggest an increase in the soil carbon pool of 9% over a 50-year timescale. -- Highlights: •Ammonia from a poultry unit can be detected 2.8 km upwind. •Ammonia caused exceedance of critical levels 800 m and critical loads 2.8 km upwind. •Dune grassland species utilised ammonia as a nutrient source. •Plant biomass increased at low levels of ammonia and total nitrogen deposition. •Soil C pools are predicted to increase by 9% over 50 years due to the excess ammonia. -- Ammonia from a poultry unit has upwind impacts, exceeding critical levels 800 m and critical loads 2.8 km upwind, and increasing biomass and tissue N of dune grassland species

  20. Procedures for the synthesis of ethylenediamine bisborane and ammonia borane

    Science.gov (United States)

    Ramachandran, Padi Veeraraghavan; Gagare, Pravin D.; Mistry, Hitesh; Biswas, Bidyut

    2017-01-03

    A method for synthesizing ammonia borane includes (a) preparing a reaction mixture in one or more solvents, the reaction mixture containing sodium borohydride, at least one ammonium salt, and ammonia; and (b) incubating the reaction mixture at temperatures between about 0.degree. C. to about room temperature in an ambient air environment under conditions sufficient to form ammonia borane. Methods for synthesizing ethylenediamine bisborane, and methods for dehydrogenation of ethylenediamine bisborane are also described.

  1. Effect of atmospheric ammonia on laying hen performance

    Energy Technology Data Exchange (ETDEWEB)

    Deaton, J.W.; Reece, F.N.; Lott, B.D.

    1982-09-01

    In periods of extremely cold weather, energy conservation in a pit-type laying house usually results in a restricted ventilation rate and an increase in air pollutants particularly ammonia. Results show that 200 ppm ammonia for 17 days causes a significant loss in percent egg production and the hens lose a significant amount of weight with a reduced feed intake. Although not satisfactory, it appears that lesser amounts of ammonia (100 ppm) can be tolerated for short periods without an immediate drastic loss in laying performance if a choice has to be made between frozen waterers and cold stress versus atmospheric ammonia in the laying house.

  2. Ammonia transport in the kidney by Rhesus glycoproteins

    Science.gov (United States)

    Verlander, Jill W.

    2014-01-01

    Renal ammonia metabolism is a fundamental element of acid-base homeostasis, comprising a major component of both basal and physiologically altered renal net acid excretion. Over the past several years, a fundamental change in our understanding of the mechanisms of renal epithelial cell ammonia transport has occurred, replacing the previous model which was based upon diffusion equilibrium for NH3 and trapping of NH4+ with a new model in which specific and regulated transport of both NH3 and NH4+ across renal epithelial cell membranes via specific membrane proteins is required for normal ammonia metabolism. A major advance has been the recognition that members of a recently recognized transporter family, the Rhesus glycoprotein family, mediate critical roles in renal and extrarenal ammonia transport. The erythroid-specific Rhesus glycoprotein, Rh A Glycoprotein (Rhag), was the first Rhesus glycoprotein recognized as an ammonia-specific transporter. Subsequently, the nonerythroid Rh glycoproteins, Rh B Glycoprotein (Rhbg) and Rh C Glycoprotein (Rhcg), were cloned and identified as ammonia transporters. They are expressed in specific cell populations and membrane domains in distal renal epithelial cells, where they facilitate ammonia secretion. In this review, we discuss the distribution of Rhbg and Rhcg in the kidney, the regulation of their expression and activity in physiological disturbances, the effects of genetic deletion on renal ammonia metabolism, and the molecular mechanisms of Rh glycoprotein-mediated ammonia transport. PMID:24647713

  3. Release of ammonia from HAN-type PHA

    International Nuclear Information System (INIS)

    Zamecnik, J.R.

    1992-01-01

    A preliminary design basis for ammonia scrubbers in the DWPF has been issued. This design basis is based on a theoretical model of ammonia evolution from the SRAT, SME and RCT. It is desirable to acquire actual process data on ammonia evolution prior to performing detailed design of scrubbers for DWPF. The evolution of ammonia from the SRAT and SME in the Integrated DWPF Melter System (IDMS) was investigated during the HM4 run. In this run, Precipitate Hydrolysis Aqueous (PHA), which was made in the Precipitate Hydrolysis Experimental Facility (PHEF) using the HAN (hydroxylamine nitrate) process was used, thus resulting in PHA with a high concentration of ammonium ion

  4. Procedure for the in vitro determination of ammonia in blood

    International Nuclear Information System (INIS)

    Jung, K.; Faust, H.

    1985-01-01

    The invention has been aimed at a procedure for the in vitro determination of ammonia in blood based on microdiffusion and isotope dilution analysis. A definite quantity of blood serum has been mixed with a definite quantity of ammonium chloride labelled with the stable isotope 15 N. The mixture has been subjected to the microdiffusion (K 2 CO 3 additives and absorption of released ammonia in sulphuric acid). The ammonia bound in sulphuric acid has been devoted to a isotope analysis and the ammonia concentration of blood has been determined according to the isotope dilution analysis equation

  5. Ammonia pollution characteristics of centralized drinking water sources in China.

    Science.gov (United States)

    Fu, Qing; Zheng, Binghui; Zhao, Xingru; Wang, Lijing; Liu, Changming

    2012-01-01

    The characteristics of ammonia in drinking water sources in China were evaluated during 2005-2009. The spatial distribution and seasonal changes of ammonia in different types of drinking water sources of 22 provinces, 5 autonomous regions and 4 municipalities were investigated. The levels of ammonia in drinking water sources follow the order of river > lake/reservoir > groundwater. The levels of ammonia concentration in river sources gradually decreased from 2005 to 2008, while no obvious change was observed in the lakes/reservoirs and groundwater drinking water sources. The proportion of the type of drinking water sources is different in different regions. In river drinking water sources, the ammonia level was varied in different regions and changed seasonally. The highest value and wide range of annual ammonia was found in South East region, while the lowest value was found in Southwest region. In lake/reservoir drinking water sources, the ammonia levels were not varied obviously in different regions. In underground drinking water sources, the ammonia levels were varied obviously in different regions due to the geological permeability and the natural features of regions. In the drinking water sources with higher ammonia levels, there are enterprises and wastewater drainages in the protected areas of the drinking water sources.

  6. Ammonia - Did it have a role in chemical evolution. [abiogenesis

    Science.gov (United States)

    Ferris, J. P.; Nicodem, D. E.

    1974-01-01

    The significance of ammonia in the chemical evolution related to the origin of life is evaluated. A computer program was employed to calculate the time needed for the decomposition of ammonia by means of a photochemical reaction. Various possible protection mechanisms for ammonia are discussed, giving attention to hydrogen sulfide, hydrogen, ozone, and CO. It is concluded that in the absence of a sufficiently high pressure of hydrogen, any ammonia present in the primitive atmosphere would have been decomposed by photolysis in a million years.

  7. Stability of hydroperoxide lyase activity from Amaranthus tricolor (Amaranthus mangostanus L.) leaves: influence of selected additives.

    Science.gov (United States)

    Long, Zhen; Kong, Xiangzhen; Zhang, Caimeng; Hua, Yufei

    2010-04-15

    Hydroperoxide lyase (HPL) has potential value for the flavour additive industry. Currently, the production and application of HPL suffer from stability problems. The objective of this study was to investigate the stabilisation of HPL preparation from Amaranthus tricolor leaves by the addition of selected chemical additives. Amaranthus tricolor leaves were identified as a particularly rich source of 13-HPL activity. The addition of 100 g L(-1) sucrose and trehalose to microsomal HPL prior to lyophilisation could retain nearly 100% enzymatic activity, compared to only 20% for the lyophilised control. The lyophilised microsomal HPL containing sucrose maintained full activity for even 40 days storage at -20 degrees C. For HPL solution, glycerol was effective for long-term stability at -20 degrees C. Moreover, poyols (sucrose and trehalose) and amino acid (glycine) enhanced the thermostability of HPL, while KCl and polyol mannitol decreased the thermostability of HPL. The flavour-producing enzyme HPL, found in the leaves of Amaranthus tricolor, was stabilised by the addition of chemical additives. (c) 2010 Society of Chemical Industry.

  8. An acidic pectin lyase from Aspergillus niger with favourable efficiency in fruit juice clarification.

    Science.gov (United States)

    Xu, S X; Qin, X; Liu, B; Zhang, D Q; Zhang, W; Wu, K; Zhang, Y H

    2015-02-01

    The pectin lyase gene pnl-zj5a from Aspergillus niger ZJ5 was identified and expressed in Pichia pastoris. PNL-ZJ5A was purified by ultrafiltration, anion exchange and gel chromatography. The Km and Vmax values determined using citrus pectin were 0.66 mg ml(-1) and 32.6 μmol min(-1) mg(-1) , respectively. PNL-ZJ5A exhibited optimal activity at 43°C and retained activity over 25-50°C. PNL-ZJ5A was optimally active at pH 5 and effective in apple juice clarification. Compared with controls, PNL-ZJ5A increased the fruit juice yield significantly. Furthermore, PNL-ZJ5A reduced the viscosity of apple juice by 38.8% and increased its transmittance by 86.3%. PNL-ZJ5A combined with a commercial pectin esterase resulted in higher juice volume. © 2014 The Society for Applied Microbiology.

  9. Potential Inhibitors for Isocitrate Lyase of Mycobacterium tuberculosis and Non-M. tuberculosis: A Summary

    Directory of Open Access Journals (Sweden)

    Yie-Vern Lee

    2015-01-01

    Full Text Available Isocitrate lyase (ICL is the first enzyme involved in glyoxylate cycle. Many plants and microorganisms are relying on glyoxylate cycle enzymes to survive upon downregulation of tricarboxylic acid cycle (TCA cycle, especially Mycobacterium tuberculosis (MTB. In fact, ICL is a potential drug target for MTB in dormancy. With the urge for new antitubercular drug to overcome tuberculosis treat such as multidrug resistant strain and HIV-coinfection, the pace of drug discovery has to be increased. There are many approaches to discovering potential inhibitor for MTB ICL and we hereby review the updated list of them. The potential inhibitors can be either a natural compound or synthetic compound. Moreover, these compounds are not necessary to be discovered only from MTB ICL, as it can also be discovered by a non-MTB ICL. Our review is categorized into four sections, namely, (a MTB ICL with natural compounds; (b MTB ICL with synthetic compounds; (c non-MTB ICL with natural compounds; and (d non-MTB ICL with synthetic compounds. Each of the approaches is capable of overcoming different challenges of inhibitor discovery. We hope that this paper will benefit the discovery of better inhibitor for ICL.

  10. Activation and Stabilization of Olive Recombinant 13-Hydroperoxide Lyase Using Selected Additives.

    Science.gov (United States)

    Jacopini, Sabrina; Vincenti, Sophie; Mariani, Magali; Brunini-Bronzini de Caraffa, Virginie; Gambotti, Claude; Desjobert, Jean-Marie; Muselli, Alain; Costa, Jean; Tomi, Félix; Berti, Liliane; Maury, Jacques

    2017-07-01

    The stabilization of olive recombinant hydroperoxide lyases (rHPLs) was investigated using selected chemical additives. Two rHPLs were studied: HPL full-length and HPL with its chloroplast transit peptide deleted (matured HPL). Both olive rHPLs are relatively stable at 4 °C, and enzyme activity can be preserved (about 100% of the rHPL activities are maintained) during 5 weeks of storage at -20 or at -80 °C in the presence of glycerol (10%, v/v). Among the additives used in this study, glycine (2.5% w/v), NaCl (0.5 M), and Na 2 SO 4 (0.25 M) provided the highest activation of HPL full-length activity, while the best matured HPL activity was obtained with Na 2 SO 4 (0.25 M) and NaCl (1 M). Although the inactivation rate constants (k) showed that these additives inactivate both rHPLs, their use is still relevant as they strongly increase HPL activity. Results of C6-aldehyde production assays also showed that glycine, NaCl, and Na 2 SO 4 are appropriate additives and that NaCl appears to be the best additive, at least for hexanal production.

  11. ATP-Citrate Lyase Controls a Glucose-to-Acetate Metabolic Switch

    Directory of Open Access Journals (Sweden)

    Steven Zhao

    2016-10-01

    Full Text Available Mechanisms of metabolic flexibility enable cells to survive under stressful conditions and can thwart therapeutic responses. Acetyl-coenzyme A (CoA plays central roles in energy production, lipid metabolism, and epigenomic modifications. Here, we show that, upon genetic deletion of Acly, the gene coding for ATP-citrate lyase (ACLY, cells remain viable and proliferate, although at an impaired rate. In the absence of ACLY, cells upregulate ACSS2 and utilize exogenous acetate to provide acetyl-CoA for de novo lipogenesis (DNL and histone acetylation. A physiological level of acetate is sufficient for cell viability and abundant acetyl-CoA production, although histone acetylation levels remain low in ACLY-deficient cells unless supplemented with high levels of acetate. ACLY-deficient adipocytes accumulate lipid in vivo, exhibit increased acetyl-CoA and malonyl-CoA production from acetate, and display some differences in fatty acid content and synthesis. Together, these data indicate that engagement of acetate metabolism is a crucial, although partial, mechanism of compensation for ACLY deficiency.

  12. Adenylosuccinate lyase (ADSL) and infantile autism: Absence of previously reported point mutation

    Energy Technology Data Exchange (ETDEWEB)

    Fon, E.A.; Sarrazin, J.; Rouleau, G.A. [Montreal General Hospital (Canada)] [and others

    1995-12-18

    Autism is a heterogeneous neuropsychiatric syndrome of unknown etiology. There is evidence that a deficiency in the enzyme adenylosuccinate lyase (ADSL), essential for de novo purine biosynthesis, could be involved in the pathogenesis of certain cases. A point mutation in the ADSL gene, resulting in a predicted serine-to-proline substitution and conferring structural instability to the mutant enzyme, has been reported previously in 3 affected siblings. In order to determine the prevalence of the mutation, we PCR-amplified the exon spanning the site of this mutation from the genomic DNA of patients fulfilling DSM-III-R criteria for autistic disorder. None of the 119 patients tested were found to have this mutation. Furthermore, on preliminary screening using single-strand conformation polymorphism (SSCP), no novel mutations were detected in the coding sequence of four ADSL exons, spanning approximately 50% of the cDNA. In light of these findings, it appears that mutations in the ADSL gene represent a distinctly uncommon cause of autism. 12 refs., 2 figs.

  13. Over-expression of hydroxynitrile lyase in transgenic cassava roots accelerates cyanogenesis and food detoxification.

    Science.gov (United States)

    Siritunga, Dimuth; Arias-Garzon, Diana; White, Wanda; Sayre, Richard T

    2004-01-01

    Cassava (Manihot esculenta, Crantz) roots are the primary source of calories for more than 500 million people, the majority of whom live in the developing countries of Africa. Cassava leaves and roots contain potentially toxic levels of cyanogenic glycosides. Consumption of residual cyanogens (linamarin or acetone cyanohydrin) in incompletely processed cassava roots can cause cyanide poisoning. Hydroxynitrile lyase (HNL), which catalyses the conversion of acetone cyanohydrin to cyanide, is expressed predominantly in the cell walls and laticifers of leaves. In contrast, roots have very low levels of HNL expression. We have over-expressed HNL in transgenic cassava plants under the control of a double 35S CaMV promoter. We show that HNL activity increased more than twofold in leaves and 13-fold in roots of transgenic plants relative to wild-type plants. Elevated HNL levels were correlated with substantially reduced acetone cyanohydrin levels and increased cyanide volatilization in processed or homogenized roots. Unlike acyanogenic cassava, transgenic plants over-expressing HNL in roots retain the herbivore deterrence of cyanogens while providing a safer food product.

  14. Purification and characterization of selenocysteine beta-lyase from Citrobacter freundii

    International Nuclear Information System (INIS)

    Chocat, P.; Esaki, N.; Tanizawa, K.; Nakamura, K.; Tanaka, H.; Soda, K.

    1985-01-01

    The purification and characterization of bacterial selenocysteine beta-lyase, an enzyme which specifically catalyzes the cleavage of L-selenocysteine to L-alanine and Se0, are presented. The enzyme, purified to near homogeneity from Citrobacter freundii, is monomeric with a molecular weight of ca. 64,000 and contains 1 mol of pyridoxal 5'-phosphate as a cofactor per mol of enzyme. L-Selenocysteine is the sole substrate. L-Cysteine is a competitive inhibitor of the enzyme. The enzyme also catalyzes the alpha, beta elimination of beta-chloro-L-alanine to form NH 3 , pyruvate, and Cl- and is irreversibly inactivated during the reaction. The physicochemical properties, e.g., amino acid composition and subunit structure, of the bacterial enzyme are fairly different from those of the pig liver enzyme. However, the catalytic properties of both enzymes, e.g., substrate specificity and inactivation by the substrate or a mechanism-based inactivator, beta-chloro-L-alanine, are very similar

  15. A high-throughput scintillation proximity assay for sphingosine-1-phosphate lyase.

    Science.gov (United States)

    Kashem, Mohammed A; Wa, Chunling; Wolak, John P; Grafos, Nicholas S; Ryan, Kelli R; Sanville-Ross, Mary L; Fogarty, Kylie E; Rybina, Irina V; Shoultz, Alycia; Molinaro, Teresa; Desai, Sudha N; Rajan, Anusha; Huber, John D; Nelson, Richard M

    2014-06-01

    The emergence of sphingosine-1-phosphate lyase (SPL) as a promising therapeutic target for inflammatory diseases has heightened interest in the identification of small molecules that modulate its activity. The enzymatic activity of SPL is typically measured using radiometric or fluorescence-based assays that require a lipid extraction step, or by direct quantitation of reaction products using mass spectrometry (MS). To facilitate testing large numbers of compounds to identify SPL modulators, we developed a robust scintillation proximity assay (SPA) that is compatible with high-throughput screening (HTS). This assay employs recombinant human full-length SPL in insect cell membrane preparations to catalyze the conversion of biotinylated aminosphingosine-1-[(33)P]phosphate (S1(33)P-biotin) to trans-2-hexadecenal-biotin and ethanolamine [(33)P]phosphate. To validate the SPA and confirm the fidelity of its measurement of SPL enzyme activity, we developed a Rapid-Fire MS method that quantitates nonradiolabeled S1P-biotin. In addition, we developed a simple, scalable method to produce S1(33)P-biotin in quantities sufficient for HTS. The optimized SPA screen in 384-well microplates produced a mean plate-wise Z'-statistic of 0.58 across approximately 3,000 plates and identified several distinct structural classes of SPL inhibitor. Among the inhibitors that the screen identified was one compound with an IC50 of 1.6 μM in the SPA that induced dose-dependent lymphopenia in mice.

  16. Diagnosis of adenylosuccinate lyase deficiency by metabolomic profiling in plasma reveals a phenotypic spectrum

    Directory of Open Access Journals (Sweden)

    Taraka R. Donti

    2016-09-01

    Full Text Available Adenylosuccinate lyase (ADSL deficiency is a rare autosomal recessive neurometabolic disorder that presents with a broad-spectrum of neurological and physiological symptoms. The ADSL gene produces an enzyme with binary molecular roles in de novo purine synthesis and purine nucleotide recycling. The biochemical phenotype of ADSL deficiency, accumulation of SAICAr and succinyladenosine (S-Ado in biofluids of affected individuals, serves as the traditional target for diagnosis with targeted quantitative urine purine analysis employed as the predominate method of detection. In this study, we report the diagnosis of ADSL deficiency using an alternative method, untargeted metabolomic profiling, an analytical scheme capable of generating semi-quantitative z-score values for over 1000 unique compounds in a single analysis of a specimen. Using this method to analyze plasma, we diagnosed ADSL deficiency in four patients and confirmed these findings with targeted quantitative biochemical analysis and molecular genetic testing. ADSL deficiency is part of a large a group of neurometabolic disorders, with a wide range of severity and sharing a broad differential diagnosis. This phenotypic similarity among these many inborn errors of metabolism (IEMs has classically stood as a hurdle in their initial diagnosis and subsequent treatment. The findings presented here demonstrate the clinical utility of metabolomic profiling in the diagnosis of ADSL deficiency and highlights the potential of this technology in the diagnostic evaluation of individuals with neurologic phenotypes.

  17. PECTATE LYASE-LIKE 9 from Brassica campestris is associated with intine formation.

    Science.gov (United States)

    Jiang, Jingjing; Yao, Lina; Yu, Youjian; Liang, Ying; Jiang, Jianxia; Ye, Nenghui; Miao, Ying; Cao, Jiashu

    2014-12-01

    Brassica campestris pectate lyase-like 9 (BcPLL9) was previously identified as a differentially expressed gene both in buds during late pollen developmental stage and in pistils during fertilization in Chinese cabbage. To characterize the gene's function, antisense-RNA lines of BcPLL9 (bcpll9) were constructed in Chinese cabbage. Self- and cross-fertilization experiments harvested half seed yields when bcpll9 lines were used as pollen donors. In vivo and in vitro pollen germination assays showed that nearly half of the pollen tubes in bcpll9 were irregular with shorter length and uneven surface. Aniline blue staining identified abnormal accumulation of a specific bright blue unknown material in the bcpll9 pollen portion. Scanning electron microscopy observation verified the abnormal outthrust material to be near the pollen germinal furrows. Transmission electron microscopy observation revealed the internal endintine layer was overdeveloped and predominantly occupied the intine. This abnormally formed intine likely induced the wavy structure and growth arrest of the pollen tube in half of the bcpll9 pollen grains, which resulted in less seed yields. Collectively, this study presented a novel PLL gene that has an important function in B. campestris intine formation. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  18. PECTATE LYASE-LIKE10 is associated with pollen wall development in Brassica campestris.

    Science.gov (United States)

    Jiang, Jingjing; Yao, Lina; Yu, Youjian; Lv, Meiling; Miao, Ying; Cao, Jiashu

    2014-11-01

    PECTATE LYASE-LIKE10 (PLL10) was previously identified as one of the differentially expressed genes both in microspores during the late pollen developmental stages and in pistils during the fertilization process in Chinese cabbage (Brassica campestris ssp. chinensis). Here, antisense-RNA was used to study the functions of BcPLL10 in Chinese cabbage. Abnormal pollen was identified in the transgenic lines (bcpll10-4, -5, and -6). In fertilization experiments, fewer seeds were harvested when the antisense-RNA lines were used as pollen donor. In vivo and in vitro pollen germination assays less germinated pollen tubes were observed in bcpll10 lines. Scanning electron microscopy observation verified that the tryphine materials were over accumulated around the pollen surface and sticked them together in bcpll10. Moreover, transmission electron microscopy observation revealed that the internal endintine was overdeveloped and predominantly occupied the intine, and disturbed the normal proportional distribution of the two layers in the non-germinal furrow region; and no obvious demarcation existed between them in the germinal furrow region in the bcpll10 pollen. Collectively, this study presented a novel PLL gene that played an important role during the pollen wall development in B. campestris, which may also possess potential importance for male sterility usage in agriculture. © 2014 Institute of Botany, Chinese Academy of Sciences.

  19. Pectin lyase overproduction by Penicillium griseoroseum mutants resistant to catabolite repression

    Directory of Open Access Journals (Sweden)

    Juliana Oliveira Lima

    Full Text Available Abstract Expression of pectinolytic genes is regulated by catabolic repression limiting the production of pectin lyase (PL if the natural inducer, pectin, is missing from the growth medium. Here, we report the isolation of Penicillium griseoroseum mutants resistant to 2-deoxy-d-glucose (DG that show resistance to catabolite repression and overproduce PL. Three spontaneous and nine UV-induced mutants were obtained. Some mutants produced sectors (segments morphologically different that were also studied. The mutants were analyzed for pectinases production on pectinase-agar plates and five mutants and two sectors showing larger clearing zones than the wild type were selected for quantitative assay. Although PL production higher than the wild type has been found, phenotype instability was observed for most of the mutants and, after transfers to nonselective medium, the DG resistance was no longer present. Only mutants M03 and M04 were stable maintaining the DG-resistance phenotype. When growing for 120 h in liquid medium containing glucose with or without pectin, both mutants showed higher PL production. In the presence of glucose as sole carbon source, the mutant M03 produced 7.8-fold more PL than the wild type. Due its phenotypic stability and PL overproduction, the mutant M03 presents potential for industrial applications.

  20. Revised domain structure of ulvan lyase and characterization of the first ulvan binding domain.

    Science.gov (United States)

    Melcher, Rebecca L J; Neumann, Marten; Fuenzalida Werner, Juan Pablo; Gröhn, Franziska; Moerschbacher, Bruno M

    2017-03-22

    Biomass waste products from green algae have recently been given new life, as these polysaccharides have potential applications in industry, agriculture, and medicine. One such polysaccharide group called ulvans displays many different, potentially useful properties that arise from their structural versatility. Hence, performing structural analyses on ulvan is crucial for future applications. However, chemical reaction-based analysis methods cannot fully characterize ulvan and tend to alter its structure. Thus, better methods require well-characterized ulvan-degrading enzymes. Therefore, we analysed a previously sequenced ulvan lyase (Genebank TM reference number JN104480) and characterized its domains. We suggest that the enzyme consists of a shorter than previously described catalytic domain, a newly identified substrate binding domain, and a C-terminal type 9 secretion system signal peptide. By separately expressing the two domains in E. coli, we confirmed that the binding domain is ulvan specific, having higher affinity for ulvan than most lectins for their ligands (affinity constant: 10 5  M -1 ). To our knowledge, this is the first description of an ulvan-binding domain. Overall, identifying this new binding domain is one step towards engineering ulvan enzymes that can be used to characterize ulvan, e.g. through enzymatic/mass spectrometric fingerprinting analyses, and help unlock its full potential.

  1. Enzymatic Hydrolysis of Alginate to Produce Oligosaccharides by a New Purified Endo-Type Alginate Lyase

    Directory of Open Access Journals (Sweden)

    Benwei Zhu

    2016-06-01

    Full Text Available Enzymatic hydrolysis of sodium alginate to produce alginate oligosaccharides has drawn increasing attention due to its advantages of containing a wild reaction condition, excellent gel properties and specific products easy for purification. However, the efficient commercial enzyme tools are rarely available. A new alginate lyase with high activity (24,038 U/mg has been purified from a newly isolated marine strain, Cellulophaga sp. NJ-1. The enzyme was most active at 50 °C and pH 8.0 and maintained stability at a broad pH range (6.0–10.0 and temperature below 40 °C. It had broad substrate specificity toward sodium alginate, heteropolymeric MG blocks (polyMG, homopolymeric M blocks (polyM and homopolymeric G blocks (polyG, and possessed higher affinity toward polyG (15.63 mM as well as polyMG (23.90 mM than polyM (53.61 mM and sodium alginate (27.21 mM. The TLC and MS spectroscopy analysis of degradation products suggested that it completely hydrolyzed sodium alginate into oligosaccharides of low degrees of polymerization (DPs. The excellent properties would make it a promising tool for full use of sodium alginate to produce oligosaccharides.

  2. Attenuated adenylosuccinate lyase deficiency: a report of one case and a review of the literature.

    Science.gov (United States)

    Jurecka, Agnieszka; Zikanova, Marie; Jurkiewicz, Elżbieta; Tylki-Szymańska, Anna

    2014-02-01

    We present a 9-year follow-up of a patient with an attenuated (type II) adenylosuccinate lyase deficiency with no obvious signs of disease progression and degradation. We also review the literature, focusing on attenuated phenotype, and we report a positive effect of a ketogenic diet on seizure control. The patient presented at the age of 5 months with a history of global developmental delay. Screening of urinary purine metabolites revealed elevation of succinyladenosine and succinylaminoimidazolecarboxamide riboside (a ratio of 2:1). Mutation analysis revealed a compound heterozygosity for missense mutations: p.R426H and p.D268H. She began to walk independently at the age of 3 years. From the age of 4 years, her communication skills improved and she presented fewer autistic features. Due to poor results in seizure control, the ketogenic diet was introduced at the age of 7 years, resulting in reduction of seizure frequency. Currently, at the age of 9 years, the girl is attending a special kindergarten and is functioning very well in her preschool group. She began to make statements that form a logical continuity and make progress in simple manual operations. The patient participates in therapies such as pet therapy, hippotherapy, speech therapy, physiotherapy, hydrotherapy, and music therapy. Georg Thieme Verlag KG Stuttgart · New York.

  3. Adenylosuccinate lyase (ADSL) and infantile autism: absence of previously reported point mutation.

    Science.gov (United States)

    Fon, E A; Sarrazin, J; Meunier, C; Alarcia, J; Shevell, M I; Philippe, A; Leboyer, M; Rouleau, G A

    1995-12-18

    Autism is a heterogeneous neuropsychiatric syndrome of unknown etiology. There is evidence that a deficiency in the enzyme adenylosuccinate lyase (ADSL), essential for de novo purine biosynthesis, could be involved in the pathogenesis of certain cases. A point mutation in the ADSL gene, resulting in a predicted serine-to-proline substitution and conferring structural instability to the mutant enzyme, has been reported previously in 3 affected siblings. In order to determine the prevalence of the mutation, we PCR-amplified the exon spanning the site of this mutation from the genomic DNA of patients fulfilling DSM-III-R criteria for autistic disorder. None of the 119 patients tested were found to have this mutation. Furthermore, on preliminary screening using singlestrand conformation polymorphism (SSCP), no novel mutations were detected in the coding sequence of four ADSL exons, spanning approximately 50% of the cDNA. In light of these findings, it appears that mutations in the ADSL gene represent a distinctly uncommon cause of autism.

  4. In vivo proton MR spectroscopy findings specific for adenylosuccinate lyase deficiency.

    Science.gov (United States)

    Henneke, M; Dreha-Kulaczewski, S; Brockmann, K; van der Graaf, M; Willemsen, M A A P; Engelke, U; Dechent, P; Heerschap, A; Helms, G; Wevers, R A; Gärtner, J

    2010-06-01

    Adenylosuccinate lyase (ADSL) deficiency is an inherited metabolic disorder affecting predominantly the central nervous system. The disease is characterized by the accumulation of succinylaminoimidazolecarboxamide riboside and succinyladenosine (S-Ado) in tissue and body fluids. Three children presented with muscular hypotonia, psychomotor delay, behavioral abnormalities, and white matter changes on brain MRI. Two of them were affected by seizures. Screening for inborn errors of metabolism including in vitro high resolution proton MRS revealed an ADSL deficiency that was confirmed genetically in all cases. All patients were studied by in vivo proton MRS. In vitro high resolution proton MRS of patient cerebrospinal fluid showed singlet resonances at 8.27 and 8.29 ppm that correspond to accumulated S-Ado. In vivo proton MRS measurements also revealed a prominent signal at 8.3 ppm in gray and white matter brain regions of all patients. The resonance was undetectable in healthy human brain. In vivo proton MRS provides a conclusive finding in ADSL deficiency and represents a reliable noninvasive diagnostic tool for this neurometabolic disorder.

  5. Intrafamilial variability in the phenotypic expression of adenylosuccinate lyase deficiency: a report on three patients.

    Science.gov (United States)

    Edery, Patrick; Chabrier, Stéphane; Ceballos-Picot, Irène; Marie, Sandrine; Vincent, Marie-Françoise; Tardieu, Marc

    2003-07-15

    We report on the striking variable expression of adenylosuccinate lyase (ADSL) deficiency in three patients belonging to a family which originates from Portugal. ADSL deficiency is a rare autosomal recessive disorder of the de novo purine synthesis which results in accumulation of succinylpurines in body fluids. As a result, patients may have variable combinations of psychomotor retardation and/or regression, seizures, autistic features and cerebellar vermis hypoplasia. However, intrafamilial variable expression of the phenotype has not been documented to date in this disease and is not commonly observed in metabolic disorders. Here, while the proband had marked psychomotor regression and progressive cerebellar vermis atrophy, the other two affected patients presented mainly autistic features. Mutation analysis of the ADSL gene revealed the presence of a homozygous R426H mutation in this family. Finally, although ADSL deficiency is a rare disorder, this diagnosis should be considered and assessed using a simple urinary screening method for the presence of succinylpurines in any patient with mental retardation of unexplained origin. Copyright 2003 Wiley-Liss, Inc.

  6. D-ribose therapy in four Polish patients with adenylosuccinate lyase deficiency: absence of positive effect.

    Science.gov (United States)

    Jurecka, A; Tylki-Szymanska, A; Zikanova, M; Krijt, J; Kmoch, S

    2008-12-01

    Deficiency of adenylosuccinate lyase (ADSL) (OMIM 103050) is an autosomal recessive disorder of the purine de novo synthesis pathway and purine nucleotide cycle, diagnosed so far in approximately 50 patients. The clinical presentation is characterized by severe neurological involvement including hypotonia, seizures, developmental delay and autistic features. Epilepsy in ADSL deficiency is frequent and occurs in approximately two-thirds of patients, beginning either early in the neonatal period or after the first year of life. At present there is no treatment of proven clinical efficacy. Despite of the increasing number of ADSL-deficient patients reported, there are only a few communications of therapeutic considerations or efforts. Among them only two showed some beneficial effects in ADSL-deficient patients. D-ribose, a simple and relatively cheap therapy, has been associated with improvement of behaviour and progressive reduction of the seizure frequency in one 13-year-old patient with ADSL deficiency. In this study we have re-examined D-ribose treatment in four ADSL-deficient patients. Assessments consisted of biochemical markers and neurological outcome. The 12-month trial of D-ribose failed to show any clinical benefit in ADSL patients with both milder and severe phenotype. D-ribose administration was accompanied by neither reduction in seizure frequency nor growth enhancement. Additionally, patients with milder type II presented the first seizure after 4 and 8 months of the D-ribose treatment. Therefore, we could not confirm a positive effect of D-ribose as previously reported.

  7. Diagnosis of adenylosuccinate lyase deficiency by metabolomic profiling in plasma reveals a phenotypic spectrum.

    Science.gov (United States)

    Donti, Taraka R; Cappuccio, Gerarda; Hubert, Leroy; Neira, Juanita; Atwal, Paldeep S; Miller, Marcus J; Cardon, Aaron L; Sutton, V Reid; Porter, Brenda E; Baumer, Fiona M; Wangler, Michael F; Sun, Qin; Emrick, Lisa T; Elsea, Sarah H

    2016-09-01

    Adenylosuccinate lyase (ADSL) deficiency is a rare autosomal recessive neurometabolic disorder that presents with a broad-spectrum of neurological and physiological symptoms. The ADSL gene produces an enzyme with binary molecular roles in de novo purine synthesis and purine nucleotide recycling. The biochemical phenotype of ADSL deficiency, accumulation of SAICAr and succinyladenosine (S-Ado) in biofluids of affected individuals, serves as the traditional target for diagnosis with targeted quantitative urine purine analysis employed as the predominate method of detection. In this study, we report the diagnosis of ADSL deficiency using an alternative method, untargeted metabolomic profiling, an analytical scheme capable of generating semi-quantitative z-score values for over 1000 unique compounds in a single analysis of a specimen. Using this method to analyze plasma, we diagnosed ADSL deficiency in four patients and confirmed these findings with targeted quantitative biochemical analysis and molecular genetic testing. ADSL deficiency is part of a large a group of neurometabolic disorders, with a wide range of severity and sharing a broad differential diagnosis. This phenotypic similarity among these many inborn errors of metabolism (IEMs) has classically stood as a hurdle in their initial diagnosis and subsequent treatment. The findings presented here demonstrate the clinical utility of metabolomic profiling in the diagnosis of ADSL deficiency and highlights the potential of this technology in the diagnostic evaluation of individuals with neurologic phenotypes.

  8. Clinical, biochemical and molecular findings in seven Polish patients with adenylosuccinate lyase deficiency.

    Science.gov (United States)

    Jurecka, Agnieszka; Zikanova, Marie; Tylki-Szymanska, Anna; Krijt, Jakub; Bogdanska, Anna; Gradowska, Wanda; Mullerova, Karolina; Sykut-Cegielska, Jolanta; Kmoch, Stanislav; Pronicka, Ewa

    2008-08-01

    Adenylosuccinate lyase (ADSL) catalyzes two steps in purine nucleotide metabolism-the 8th step in the de novo pathway: conversion of succinylaminoimidazole carboxamide ribotide (SAICAR) to aminoimidazole carboxamide ribotide (AICAR), and conversion of adenylosuccinate (S-AMP) to adenylate (AMP) in the purine nucleotide cycle. To date, over 50 patients have been reported suffering from ADSL deficiency. We report all seven so far diagnosed Polish patients with this defect. Most of our patients shared intractable seizures and psychomotor retardation since the neonatal period and had biochemical evidence of severe (type I) deficiency. Two patients with type II suffered only from mild/moderate psychomotor retardation and showed a transientvisual contact disturbance. One patient had a fatal neonatal form of ADSL deficiency with lack of spontaneous movement, respiratory failure, severe encephalopathy and intractable seizures. Analysis of the ADSL gene showed that four apparently unrelated patients carried a R426H mutation (two homozygous and two compound heterozygous). With the exception of the latter mutation, a Y114H mutation that had been reported previously, and a novel mutation T242I, all other mutations (including D268H and three novel S23R, D215H and I351T mutations) were found only in single families in single alleles. A search for this disorder should be included in the screening program of all infants with unexplained neonatal seizures, severe infantile epileptic encephalopathy, developmental delay, hypotonia, and/or autistic features.

  9. The Salmonella effector protein SpvC, a phosphothreonine lyase is functional in plant cells

    KAUST Repository

    Neumann, Christina

    2014-10-17

    Salmonella is one of the most prominent causes of food poisoning and growing evidence indicates that contaminated fruits and vegetables are an increasing concern for human health. Successful infection demands the suppression of the host immune system, which is often achieved via injection of bacterial effector proteins into host cells. In this report we present the function of Salmonella effector protein in plant cell, supporting the new concept of trans-kingdom competence of this bacterium. We screened a range of Salmonella Typhimurium effector proteins for interference with plant immunity. Among these, the phosphothreonine lyase SpvC attenuated the induction of immunity-related genes when present in plant cells. Using in vitro and in vivo systems we show that this effector protein interacts with and dephosphorylates activated Arabidopsis Mitogen-activated Protein Kinase 6 (MPK6), thereby inhibiting defense signaling. Moreover, the requirement of Salmonella SpvC was shown by the decreased proliferation of the ΔspvC mutant in Arabidopsis plants. These results suggest that some Salmonella effector proteins could have a conserved function during proliferation in different hosts. The fact that Salmonella and other Enterobacteriaceae use plants as hosts strongly suggests that plants represent a much larger reservoir for animal pathogens than so far estimated.

  10. Production of endo-pectate lyase by two stage cultivation of Erwinia carotovora

    Energy Technology Data Exchange (ETDEWEB)

    Fukuoka, Satoshi; Kobayashi, Yoshiaki

    1987-02-26

    The productivity of endo-pectate lyase from Erwinia carotovora GIR 1044 was found to be greatly improved by two stage cultivation: in the first stage the bacterium was grown with an inducing carbon source, e.g., pectin, and in the second stage it was cultivated with glycerol, xylose, or fructose with the addition of monosodium L-glutamate as nitrogen source. In the two stage cultivation using pectin or glycerol as the carbon source the enzyme activity reached 400 units/ml, almost 3 times as much as that of one stage cultivation in a 10 liter fermentor. Using two stage cultivation in the 200 liter fermentor improved enzyme productivity over that in the 10 liter fermentor, with 500 units/ml of activity. Compared with the cultivation in Erlenmeyer flasks, fermentor cultivation improved enzyme productivity. The optimum cultivating conditions were agitation of 480 rpm with aeration of 0.5 vvm at 28 /sup 0/C. (4 figs, 4 tabs, 14 refs)

  11. Characterization of C-S lyase from Lactobacillus delbrueckii subsp. bulgaricus ATCC BAA-365 and its potential role in food flavour applications.

    Science.gov (United States)

    Allegrini, Alessandra; Astegno, Alessandra; La Verde, Valentina; Dominici, Paola

    2017-04-01

    Volatile thiols have substantial impact on the aroma of many beverages and foods. Thus, the control of their formation, which has been linked to C-S lyase enzymatic activities, is of great significance in industrial applications involving food flavours. Herein, we have carried out a spectroscopic and functional characterization of a putative pyridoxal 5'-phosphate (PLP)-dependent C-S lyase from the lactic acid bacterium Lactobacillus delbrueckii subsp. bulgaricus ATCC BAA-365 (LDB C-S lyase). Recombinant LDB C-S lyase exists as a tetramer in solution and shows spectral properties of enzymes containing PLP as cofactor. The enzyme has a broad substrate specificity toward sulphur-containing amino acids with aminoethyl-L-cysteine and L-cystine being the most effective substrates over L-cysteine and L-cystathionine. Notably, the protein also reveals cysteine-S-conjugate β-lyase activity in vitro, and is able to cleave a cysteinylated substrate precursor into the corresponding flavour-contributing thiol, with a catalytic efficiency higher than L-cystathionine. Contrary to similar enzymes of other lactic acid bacteria however, LDB C-S lyase is not capable of α,γ-elimination activity towards L-methionine to produce methanethiol, which is a significant compound in flavour development. Based on our results, future developments can be expected regarding the flavour-forming potential of Lactobacillus C-S lyase and its use in enhancing food flavours. © The Authors 2017. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.

  12. Ammonia-dimethylchloramine system: kinetic approach in an aqueous medium and comparison with the mechanism involving in liquid ammonia

    OpenAIRE

    Stephan, Juliette; Pasquet, Véronique; Elkhatib, Mazen; Goutelle, Véronique; Delalu, Henri

    2008-01-01

    International audience; After an exhaustive study of the system ammonia-dimethylchloramine in liquid ammonia, it was interesting to compare the reactivity of this system in liquid ammonia with the same systeme in an aqueous medium. Dimethylchloramine prepared in a pure state, undergoes dehydrohalogenation in an alkaline medium: the principal products formed are N-methylmethanimine, 1,3,5-trimethylhexahydrotriazine, formaldehyde and methylamine. The kinetics of this reaction was studied by UV,...

  13. [Ammonia oxidation kinetics of ammonia oxidizer mixed culture under the conditions of O2 and trace NO2 mixed gasses].

    Science.gov (United States)

    Zhang, Dai-Jun; Zu, Bo; Ren, Hong-Yang; Zhang, Ping; Cong, Li-Ying; Yan, Qing

    2008-01-01

    The kinetics of the NO2-dependent ammonia oxidation was developed for ammonia oxidizer mixed culture when there was no molecular oxygen in the batch tests. The kinetics parameters were determined, where the half saturate coefficient of NO2 was 0.821 micromol x L(-1), inhibition coefficient of NO2 concentration was 1.721 micromol x L(-1), and the maximum ammonia oxidation rate were 0.144 mg x (mg x h)(-1). After adding the volume fraction of O2 was 2% to trace NO2, the ammonia oxidation rates increased obviously. The maximum ammonia oxidation rate, 0.198 mg x (mg x h)(-1) occurred under the condition of the mixed gasses containing the volume fraction of O2 was 2% and 50 x 10(-6) NO2. Under the condition of mixed gasses containing the volume fraction of O2 was 21% to trace NO2, the ammonia oxidation rates further increased greatly. The maximum ammonia oxidation rate, 0.477 mg x (mg x h)(-1) occurred when the volume fraction of O2 was 21% and 100 x 10(-6) NO2 in the mixed gas, which is 3 times higher than the general aerobic ammonia oxidation rate. The function for NO2 apparently to enhance ammonia oxidation was suggested. The kinetics model of ammonia oxidation under the conditions of O2 and trace NO2 mixed gasses was developed. The model was validated by the results of ammonia oxidation experiments under the conditions of the mixed gasses containing 2% O2 and trace NO2. The mechanism for NO2 to enhance ammonia oxidation under the conditions of O2 and trace NO2 mixed gasses was discussed.

  14. AMMONIA-FREE NOx CONTROL SYSTEM

    Energy Technology Data Exchange (ETDEWEB)

    Song Wu; Zhen Fan; Andrew H. Seltzer; Richard G. Herman

    2006-06-01

    This report describes a novel NOx control system that has the potential to drastically reduce cost, and enhance performance, operation and safety of power plant NOx control. The new system optimizes the burner and the furnace to achieve very low NOx levels and to provide an adequate amount of CO, and uses the CO for reducing NO both in-furnace and over a downstream AFSCR (ammonia-free selective catalytic reduction) reactor. The AF-SCR combines the advantages of the highly successful SCR technology for power plants and the TWC (three-way catalytic converter) widely used on automobiles. Like the SCR, it works in oxidizing environment of combustion flue gas and uses only base metal catalysts. Like the TWC, the AF-SCR removes NO and excess CO simultaneously without using any external reagent, such as ammonia. This new process has been studied in a development program jointed funded by the US Department of Energy and Foster Wheeler. The report outlines the experimental catalyst work performed on a bench-scale reactor, including test procedure, operating conditions, and results of various catalyst formulations. Several candidate catalysts, prepared with readily available transition metal oxides and common substrate materials, have shown over 80-90% removal for both NO and CO in oxidizing gas mixtures and at elevated temperatures. A detailed combustion study of a 400 MWe coal-fired boiler, applying computational fluid dynamics techniques to model boiler and burner design, has been carried out to investigate ways to optimize the combustion process for the lowest NOx formation and optimum CO/NO ratios. Results of this boiler and burner optimization work are reported. The paper further discusses catalyst scale-up considerations and the conceptual design of a 400 MWe size AF-SCR reactor, as well as economics analysis indicating large cost savings of the ammonia-free NOx control process over the current SCR technology.

  15. Ammonia emissions of a rotational grazing system

    Science.gov (United States)

    Voglmeier, Karl; Häni, Christoph; Jocher, Markus; Ammann, Christof

    2017-04-01

    Intensive agricultural livestock production is the main source of air pollution by ammonia (NH3). Grazing is considered to reduce emissions significantly. However, ammonia emissions measurements on pastures are very rare and most emission models base their emissions factors for grazing on studies from the 1990s, which report a large emission range from 2.7% to 13.6% of the applied total ammonia nitrogen (TAN). We present first results of the Posieux pasture experiment in 2016 where NH3 concentration and fluxes were measured during the grazing season. The applied methods include an eddy covariance system with a two channel reactive nitrogen (Nr) converter measuring in parallel the sum of oxidized Nr species and the sum of the total Nr species. The difference of the two channels corresponds to the sum of reduced Nr species. Furthermore four MiniDOAS instruments for line integrated concentration measurements without an inlet system were used. The fluxes were estimated by applying a backward Lagrangian stochastic model (bLS) to the concentration difference of paired MiniDOAS up- and downwind of a sub-plot of the field. Monitoring of dung (visual survey) and urine patch locations (with soil electrical conductivity sensor) was carried out after each grazing rotation on selected sub-plots. It helped to compute statistics of the dung/urine patch distribution on the pasture. The experimental setup and the environmental conditions resulted in high temporal and spatial dynamics of NH3 concentrations and fluxes. The calculated fluxes were used to estimate the total net emission during the grazing period. Based on the average dung/urine patch distribution on the field an emission factor for the pasture was computed and compared to results from the literature. We discuss the applicability and limitations of the two measurement systems, reconsider the main emission drivers and explain differences in the results.

  16. Exposure of lambs to atmospheric ammonia

    Energy Technology Data Exchange (ETDEWEB)

    Drummond, J.G.; Curtis, S.E.; Lewis, J.M.; Hinds, F.C.; Simon, J.

    1976-01-01

    Two trials were conducted to determine the effects of chronic exposure to atmospheric ammonia at a concentration of 75 ppm on weight gain and efficiency of feed conversion by lambs. Lambs were weighed and then randomly allotted to four dynamic air-pollutant exposure chambers. Initial weights (mean +/- SE) of lambs were 22.1 +/- .46 kg and 27.5 +/- 1.54 kg for trials 1 and 2, respectively (n=12 in both trials). Lambs in two chambers (controls) were maintained in an atmosphere of filtered room air. Lambs in the other two chambers (NH/sub 3/ -exposed) were exposed to an atmosphere of filtered air plus atmospheric ammonia at a concentration of 75 ppm. Exposure period was 28 days in both trials. Feed and water were available at all times. Lamb weight gain and feed disappearance were determined weekly. Upon termination of each trial, one lamb was randomly selected from each chamber, sacrificed, and gross and histopathologic structural changes noted. Weight gain and efficiency of feed conversion data for the two trials were combined for statistical analysis. Control lambs gained on the average .09 kg/day (.28 vs .19) more (P<.01) and consumed on the average .68 kg of feed (4.53 vs 5.21) less per kilogram of weight gain (P<.10) than did NH/sub 3/-exposed lambs. Ammonia-exposed lambs in both trials showed profuse lacrimation, severe coughing and sneezing, and profuse nasal discharge, which was bloody in some instances. Gross and histopathologic findings appeared to be qualitatively similar between treatments, but more pronounced in NH/sub 3/-exposed lambs.

  17. The importance of the ammonia purification process in ammonia-water absorption systems

    International Nuclear Information System (INIS)

    Fernandez-Seara, Jose; Sieres, Jaime

    2006-01-01

    Practical experience in working with ammonia-water absorption systems shows that the ammonia purification process is a crucial issue in order to obtain an efficient and reliable system. In this paper, the detrimental effects of the residual water content in the vapour refrigerant are described and quantified based on the system design variables that determine the effectiveness of the purification process. The study has been performed considering a single stage system with a distillation column with complete condensation. The ammonia purification effectiveness of the column is analysed in terms of the efficiencies in the stripping and rectifying sections and the reflux ratio. By varying the efficiencies from 0 to 1, systems with neither the rectifying nor stripping section, with either the rectifying or stripping section, or with both sections can be considered. The impact of the ammonia purification process on the absorption system performance is studied based on the column efficiencies and reflux ratio; and its effects on refrigerant concentration, system COP, system pressures and main system mass flow rates and concentrations are analysed. When the highest efficiency rectifying sections are used a combination of generation temperature and reflux ratio which leads to optimum COP values is found. The analysis covers different operating conditions with air and water cooled systems from refrigeration to air conditioning applications by changing the evaporation temperature. The importance of rectification in each kind of application is evaluated

  18. TES ammonia retrieval strategy and global observations of the spatial and seasonal variability of ammonia (journal)

    Science.gov (United States)

    Presently only limited sets of tropospheric ammonia (NH3) measurements in the Earth's atmosphere have been reported from satellite and surface station measurements, despite the well-documented negative impact of NH3 on the environment and human health. Presented here is a detaile...

  19. Ammonia-Free NOx Control System

    Energy Technology Data Exchange (ETDEWEB)

    Song Wu; Zhen Fan; Andrew H. Seltzer; Richard G. Herman

    2005-03-31

    Research is being conducted under United States Department of Energy (DOE) Contract DE-FC26-03NT41865 to develop a new technology to achieve very low levels of NOx emissions from pulverized coal fired boiler systems by employing a novel system level integration between the PC combustion process and the catalytic NOx reduction with CO present in the combustion flue gas. The combustor design and operating conditions will be optimized to achieve atypical flue gas conditions. This approach will not only suppress NOx generation during combustion but also further reduce NOx over a downstream catalytic reactor that does not require addition of an external reductant, such as ammonia.

  20. Ammonia-Free NOx Control System

    Energy Technology Data Exchange (ETDEWEB)

    Song Wu; Zhen Fan; Andrew H. Seltzer

    2005-06-30

    Research is being conducted under United States Department of Energy (DOE) Contract DEFC26-03NT41865 to develop a new technology to achieve very low levels of NOx emissions from pulverized coal fired boiler systems by employing a novel system level integration between the PC combustion process and the catalytic NOx reduction with CO present in the combustion flue gas. The combustor design and operating conditions will be optimized to achieve atypical flue gas conditions. This approach will not only suppress NOx generation during combustion but also further reduce NOx over a downstream catalytic reactor that does not require addition of an external reductant, such as ammonia.

  1. Nonadiabatic transitions in electrostatically trapped ammonia molecules

    International Nuclear Information System (INIS)

    Kirste, Moritz; Schnell, Melanie; Meijer, Gerard; Sartakov, Boris G.

    2009-01-01

    Nonadiabatic transitions are known to be major loss channels for atoms in magnetic traps but have thus far not been experimentally reported upon for trapped molecules. We have observed and quantified losses due to nonadiabatic transitions for three isotopologues of ammonia in electrostatic traps by comparing the trapping times in traps with a zero and a nonzero electric field at the center. Nonadiabatic transitions are seen to dominate the overall loss rate even for the present samples that are at relatively high temperatures of 30 mK. It is anticipated that losses due to nonadiabatic transitions in electric fields are omnipresent in ongoing experiments on cold molecules.

  2. Modeling and simulation of ammonia removal from purge gases of ammonia plants using a catalytic Pd-Ag membrane reactor

    International Nuclear Information System (INIS)

    Rahimpour, M.R.; Asgari, A.

    2008-01-01

    In this work, the removal of ammonia from synthesis purge gas of an ammonia plant has been investigated. Since the ammonia decomposition is thermodynamically limited, a membrane reactor is used for complete decomposition. A double pipe catalytic membrane reactor is used to remove ammonia from purge gas. The purge gas is flowing in the reaction side and is converted to hydrogen and nitrogen over nickel-alumina catalyst. The hydrogen is transferred through the Pd-Ag membrane of tube side to the shell side. A mathematical model including conservation of mass in the tube and shell side of reactor is proposed. The proposed model was solved numerically and the effects of different parameters on the rector performance were investigated. The effects of pressure, temperature, flow rate (sweep ratio), membrane thickness and reactor diameter have been investigated in the present study. Increasing ammonia conversion was observed by raising the temperature, sweep ratio and reducing membrane thickness. When the pressure increases, the decomposition is gone toward completion but, at low pressure the ammonia conversion in the outset of reactor is higher than other pressures, but complete destruction of the ammonia cannot be achieved. The proposed model can be used for design of an industrial catalytic membrane reactor for removal of ammonia from ammonia plant and reducing NO x emissions

  3. Improving The Efficiency Of Ammonia Electrolysis For Hydrogen Production

    Science.gov (United States)

    Palaniappan, Ramasamy

    Given the abundance of ammonia in domestic and industrial wastes, ammonia electrolysis is a promising technology for remediation and distributed power generation in a clean and safe manner. Efficiency has been identified as one of the key issues that require improvement in order for the technology to enter the market phase. Therefore, this research was performed with the aim of improving the efficiency of hydrogen production by finding alternative materials for the cathode and electrolyte. 1. In the presence of ammonia the activity for hydrogen evolution reaction (HER) followed the trend Rh>Pt>Ru>Ni. The addition of ammonia resulted in lower rates for HER for Pt, Ru, and Ni, which have been attributed to competition from the ammonia adsorption reaction. 2. The addition of ammonia offers insight into the role of metal-hydrogen underpotential deposition (M-Hupd) on HER kinetics. In addition to offering competition via ammonia adsorption it resulted in fewer and weaker M-Hupd bonds for all metals. This finding substantiates the theory that M-Hupd bonds favor HER on Pt electrocatalyst. However, for Rh results suggest that M-Hupd bond may hinder the HER. In addition, the presence of unpaired valence shell electrons is suggested to provide higher activity for HER in the presence of ammonia. 3. Bimetals PtxM1-x (M = Ir, Ru, Rh, and Ni) offered lower overpotentials for HER compared to the unalloyed metals in the presence of ammonia. The activity of HER in the presence of ammonia follows the trend Pt-Ir>Pt-Rh>Pt-Ru>Pt-Ni. The higher activity of HER is attributed to the synergistic effect of the alloy, where ammonia adsorbs onto the more electropositive alloying metal leaving Pt available for Hupd formation and HER to take place. Additionally, this supports the theory that the presence of a higher number of unpaired electrons favors the HER in the presence of ammonia. 4. Potassium polyacrylate (PAA-K) was successfully used as a substitute for aqueous KOH for ammonia

  4. Staging properties of potassium-ammonia ternary graphite intercalation compounds at high ammonia pressure

    Science.gov (United States)

    Qian, X. W.; Solin, S. A.

    1989-04-01

    The pressure dependence of the (00l) x-ray diffraction patterns of the ternary graphite intercalation compound K(NH3)xC24 has been studied in the range 0.5-11 kbar (for which x~4.5) using a diamond anvil cell. A special apparatus for loading the cell with liquid ammonia at room temperature has been constructed and is briefly described. In these experiments, the pressure-transmitting fluid was also an intercalant, namely ammonia. Therefore, the chemical potential of this species was linearly coupled to the applied pressure in contrast to the usual case where the pressure-transmitting fluid is chemically passive. The pressure dependences of the basal spacings and of the relative intensities of key reflections have been measured, as have the compressibilities of the stage-1 and stage-2 components of the two-phase system. Basal-spacing anomalies and anomalies in the relative intensities occur at pressures of ~3.5 and 8.0 kbar and are tentatively attributed to in-plane coordination changes in the potassium-ammonia ratio. Using thermodynamic arguments and Le Chatelier's principle we show quantitatively that a staging phase transition from pure stage-1 phase to an admixture of stage-1 and stage-2 is expected with increased pressure above 10 bar in agreement with experiment. The saturation ammonia compositions (x values) of the admixed stages are found to be 4.5 and 5.4 for the stage-1 and -2 components, respectively. This result is interpreted as evidence that the composition is not sterically limited but is determined by the binding energy of ammonia for potassium and by the perturbation to this energy from the guest-host interaction.

  5. PRETREATMENT AND FRACTIONATION OF CORN STOVER BY AMMONIA RECYCLE PERCOLATION PROCESS. (R831645)

    Science.gov (United States)

    Corn stover was pretreated with aqueous ammonia in a flow-through column reactor,a process termed as Ammonia Recycle Percolation (ARP). The aqueous ammonia causesswelling and efficient delignification of biomass at high temperatures. The ARPprocess solubilizes abou...

  6. Atmospheric cycles of nitrogen oxides and ammonia

    International Nuclear Information System (INIS)

    Bottger, A.; Ehhalt, D.H.; Gravenhorst, G.

    1981-12-01

    The atmospheric cycles of nitrogenous trace compounds for the Northern and Southern Hemispheres are discussed. Source strengths and destruction rates for the nitrogen oxides: NO, NO 2 and HNO 3 -(NOx) and ammonia (NH 3 ) are given as a function of latitude over continents and oceans. The global amounts of NOx-N and NH 3 -N produced annually in the period 1950 to 1975 (34 + 5 x one trillion g NOx-N/yr and 29 + or - 6 x one trillion g NH3-N/yr) are much less than previously assumed. Globally, natural and anthropogenic emissions are of similar magnitude. The NOx emission from anthropogenic sources is 1.5 times that from natural processes in the Northern Hemisphere, whereas in the Southern Hemisphere, it is a factor of 3 or 4 less. More than 80% of atmospheric ammonia seems to be derived from excrements of domestic animals, mostly by bulk deposition: 24 + or - 9 x one trillion g NO 3 -N/yr and 21 + or - 9 x one trillion g NH 4 +-N/yr. Another fraction may be removed by absorption on vegetation and soils

  7. Method of production of ammonia synthesis gas

    Energy Technology Data Exchange (ETDEWEB)

    1943-09-10

    In the catalytic synthesis of complicated hydrocarbons from carbon monoxide and hydrogen at normal or slightly increased pressures by the Fischer--Tropsch process, or carried out at higher pressures in some other proposals, the synthesis gas was incompletely transformed. After the conclusion of the synthesis, the residual gas, upon the separation of the liquid constituents, contained, in addition to the unreacted carbon moxoxide and hydrogen, also considerable amounts of methane, carbon dioxide, and nitrogen from the original synthesis gas. This residual gas had been used as fuel. It was, however, pure and contained no sulfur or other catalyst poisons and burning it was considered uneconomical. It was proposed to make better use of it by using it as fuel. It was, however, pure and contained no sulfur or other catalyst poisons and burning it was considered uneconomical. It was proposed to make better use of it by using it as a raw material for the production of synthesis gas by decomposing the methane present in it with steam according to the equation CH/sub 4/ + H/sub 2/O = CO + 3H/sub 2/. This conversion was to be brought about either by a return to the producers or else in special splitting units. Also, it had been found that the residual gas, possibly even in the presence of oxygen compounds, could be conveniently used for the synthesis of ammonia. Several examples of ammonia synthesis were discussed.

  8. Study of ammonia synthesis using technetium catalysts

    International Nuclear Information System (INIS)

    Spitsyn, V.I.; Mikhajlenko, I.E.; Pokrovskaya, O.V.

    1982-01-01

    A study was made on catalytic properties of technetium in ammonia synthesis reaction. The preparation of technetium catalysts on ν-Al 2 O 3 , BaTiO 3 , BaO-ν-Al 2 O 3 substrates is described. The investigation of catalytic activity of catalysts was carried out at a pressure of 1 atm. in vertical reactor with volume rate of 15000 h - 1 in the temperature range of 350-425 deg. The amount of catalyst was 0.5-1 g, the volume- 0.5 ml, the size of granules- 2-3 mm. Rate constants of ammonia synthesis reaction were calculated. Seeming activation energies of the process have meanings wihtin the limits of 40-50 kcal/mol. It was shown that with increase in concentration of Tc on BaTiO 3 the catalytic activity rises in comparison with pure Tc. The reduction of catalytic activity with increase of metal content on Al 2 O 3 begins in the limits of 3.5-6.7% Tc/ν-Al 2 O 3 . The catalyst of 5.3% Tc/4.1% Ba/ν -Al 2 O 3 compound has the maximum activity. Technetium catalysts possess the stable catalytic activity and don't requre its reduction during several months

  9. Energy efficient ammonia heat pump. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Madsen, Claus; Pijnenburg, B.; Schumann Grindorf, H. [Danish Technological Institute, Aarhus (Denmark); Christensen, Rolf [Alfa Laval, Lund (Sweden); Rasmussen, Bjarne D. [Grundfos, Bjerringbro (Denmark); Gram, S.; Fredborg Jakobsen, D. [Svedan Industri Koeleanlaeg, Greve (Denmark)

    2013-09-15

    The report describes the development of a highly effective ammonia heat pump. Heat pumps play an increasingly important role in the search for more effective use of energy in our society. Highly efficient heat pumps can contribute to reduced energy consumption and improved economy of the systems which they are a part of. An ammonia heat pump with high pressure reciprocating compressor and a novel split condenser was developed to prove potential for efficiency optimization. The split of the condenser in two parts can be utilized to obtain smaller temperature approaches and, thereby, improved heat pump efficiency at an equal heat exchanger area, when compared to the traditional solution with separate condenser and de-superheater. The split condenser design can also be exploited for heating a significant share of the total heating capacity to a temperature far above the condensing temperature. Furthermore, the prototype heat pump was equipped with a plate type evaporator combined with a U-turn separator with a minimum liquid height and a liquid pump with the purpose of creating optimum liquid circulation ratio for the highest possible heat transfer coefficients at the lowest possible pressure drop. The test results successfully confirmed the highest possible efficiency; a COP of 4.3 was obtained when heating water from 40 deg. C to 80 deg. C while operating with evaporating/condensing temperatures of +20 deg C/+73 deg C. (Author)

  10. Ammonia-Oxidizing Archaea and Bacteria Differentially Contribute to Ammonia Oxidation in Sediments from Adjacent Waters of Rushan Bay, China

    Directory of Open Access Journals (Sweden)

    Hui He

    2018-02-01

    Full Text Available Ammonia oxidation plays a significant role in the nitrogen cycle in marine sediments. Ammonia-oxidizing archaea (AOA and bacteria (AOB are the key contributors to ammonia oxidation, and their relative contribution to this process is one of the most important issues related to the nitrogen cycle in the ocean. In this study, the differential contributions of AOA and AOB to ammonia oxidation in surface sediments from adjacent waters of Rushan Bay were studied based on the ammonia monooxygenase (amoA gene. Molecular biology techniques were used to analyze ammonia oxidizers’ community characteristics, and potential nitrification incubation was applied to understand the ammonia oxidizers’ community activity. The objective was to determine the community structure and activity of AOA and AOB in surface sediments from adjacent waters of Rushan Bay and to discuss the different contributions of AOA and AOB to ammonia oxidation during summer and winter seasons in the studied area. Pyrosequencing analysis revealed that the diversity of AOA was higher than that of AOB. The majority of AOA and AOB clustered into Nitrosopumilus and Nitrosospira, respectively, indicating that the Nitrosopumilus group and Nitrosospira groups may be more adaptable in studied sediments. The AOA community was closely correlated to temperature, salinity and ammonium concentration, whereas the AOB community showed a stronger correlation with temperature, chlorophyll-a content (chla and nitrite concentration. qPCR results showed that both the abundance and the transcript abundance of AOA was consistently greater than that of AOB. AOA and AOB differentially contributed to ammonia oxidation in different seasons. AOB occupied the dominant position in mediating ammonia oxidation during summer, while AOA might play a dominant role in ammonia oxidation during winter.

  11. An infrared spectroscopy method to detect ammonia in gastric juice.

    Science.gov (United States)

    Giovannozzi, Andrea M; Pennecchi, Francesca; Muller, Paul; Balma Tivola, Paolo; Roncari, Silvia; Rossi, Andrea M

    2015-11-01

    Ammonia in gastric juice is considered a potential biomarker for Helicobacter pylori infection and as a factor contributing to gastric mucosal injury. High ammonia concentrations are also found in patients with chronic renal failure, peptic ulcer disease, and chronic gastritis. Rapid and specific methods for ammonia detection are urgently required by the medical community. Here we present a method to detect ammonia directly in gastric juice based on Fourier transform infrared spectroscopy. The ammonia dissolved in biological liquid samples as ammonium ion was released in air as a gas by the shifting of the pH equilibrium of the ammonium/ammonia reaction and was detected in line by a Fourier transform infrared spectroscopy system equipped with a gas cell for the quantification. The method developed provided high sensitivity and selectivity in ammonia detection both in pure standard solutions and in a simulated gastric juice matrix over the range of diagnostic concentrations tested. Preliminary analyses were also performed on real gastric juice samples from patients with gastric mucosal injury and with symptoms of H. pylori infection, and the results were in agreement with the clinicopathology information. The whole analysis, performed in less than 10 min, can be directly applied on the sample without extraction procedures and it ensures high specificity of detection because of the ammonia fingerprint absorption bands in the infrared spectrum. This method could be easily used with endoscopy instrumentation to provide information in real time and would enable the endoscopist to improve and integrate gastroscopic examinations.

  12. Modelling of ammonia emissions from dairy cow houses

    NARCIS (Netherlands)

    Monteny, G.J.

    2000-01-01

    Dairy cow husbandry contributes to environmental acidification through the emission of ammonia. In-depth knowledge on the processes and variable factors that play a role in the emission of ammonia from dairy cow houses benefits the production of emission data, the development of low

  13. Niche differentiation between ammonia-oxidizing bacteria in aquatic environments

    NARCIS (Netherlands)

    Coci, M.

    2007-01-01

    The aim of the studies presented in this thesis was the search for niche differentiation between the ammonia-oxidizing bacteria in aquatic environments. Ammonia-oxidizing bacteria are chemolitho-autotrophic microorganisms responsible for the first, mostly rate-limiting step of the nitrification

  14. Electro Decomposition of Ammonia into Hydrogen for Fuel Cell Use

    Science.gov (United States)

    2012-01-01

    400 million gallon per day (MGD). Currently, wastewater treatment solutions for ammonia ( biological and chemical treatments) consume a significant...of the catalytic and inhibition mechanisms of urease. Journal of Biological Inorganic Chemistry. 6(3):300-314. National Research Council (NRC) and...electrodes for urea oxidation (Talk) Denitrification of wastewater through ammonia electrolysis (Talk) Journal publicationsg Daramola, Damilola A

  15. Ammonia loss, ammonium and nitrate accumulation from mixing ...

    African Journals Online (AJOL)

    user

    2011-04-25

    Apr 25, 2011 ... Ammonia loss from urea significantly hinders efficient use of urea in agriculture. In order to reduce ammonia loss and, at the same time, improve beneficial accumulation of soil exchangeable ammonium and nitrate for efficient utilization by plants, this laboratory study was conducted to determine the effect.

  16. Electrolytic synthesis of ammonia in molten salts under atmospheric pressure.

    Science.gov (United States)

    Murakami, Tsuyoshi; Nishikiori, Tokujiro; Nohira, Toshiyuki; Ito, Yasuhiko

    2003-01-15

    Ammonia was successfully synthesized by using a new electrochemical reaction with high current efficiency at atmospheric pressure and at lower temperatures than the Haber-Bosch process. In this method, nitride ion (N3-), which is produced by the reduction from nitrogen gas at the cathode, is anodically oxidized and reacts with hydrogen to produce ammonia at the anode.

  17. Ammonia Induces Autophagy through Dopamine Receptor D3 and MTOR

    Science.gov (United States)

    Li, Zhiyuan; Ji, Xinmiao; Wang, Wenchao; Liu, Juanjuan; Liang, Xiaofei; Wu, Hong; Liu, Jing; Eggert, Ulrike S.; Liu, Qingsong

    2016-01-01

    Hyperammonemia is frequently seen in tumor microenvironments as well as in liver diseases where it can lead to severe brain damage or death. Ammonia induces autophagy, a mechanism that tumor cells may use to protect themselves from external stresses. However, how cells sense ammonia has been unclear. Here we show that culture medium alone containing Glutamine can generate milimolar of ammonia at 37 degrees in the absence of cells. In addition, we reveal that ammonia acts through the G protein-coupled receptor DRD3 (Dopamine receptor D3) to induce autophagy. At the same time, ammonia induces DRD3 degradation, which involves PIK3C3/VPS34-dependent pathways. Ammonia inhibits MTOR (mechanistic target of Rapamycin) activity and localization in cells, which is mediated by DRD3. Therefore, ammonia has dual roles in autophagy: one to induce autophagy through DRD3 and MTOR, the other to increase autophagosomal pH to inhibit autophagic flux. Our study not only adds a new sensing and output pathway for DRD3 that bridges ammonia sensing and autophagy induction, but also provides potential mechanisms for the clinical consequences of hyperammonemia in brain damage, neurodegenerative diseases and tumors. PMID:27077655

  18. Ammonia role in WWER primary circuit water chemistry optimization

    International Nuclear Information System (INIS)

    Kritskij, V.G.; Stjagkin, P.S.; Chvedova, M.N.; Slobodov, A.A.

    1999-01-01

    Ammonia influence on iron crud's solubility at 300 deg. C and different relations of boric acid and alkaline cation sum are considered. Reduction of dose rate on WWER-440 steam generators at average ammonia concentration increasing is empirically explained. Practical recommendations on optimization of WWER primary circuit water chemistry are given. (author)

  19. AMINA-chip : a miniaturized measurement system for ambient ammonia

    NARCIS (Netherlands)

    Timmer, Björn Herman

    2004-01-01

    The development of a miniaturized and integrated measurement system for gaseous ammonia is described in this thesis. The measuring principle, ¿AMINA¿, is an indirect method for selectively measuring ammonia that makes use of pH-transitions, electrolyte conductivity detection and phaseseparating

  20. Kinetics of absorption of carbon dioxide in aqueous ammonia solutions

    NARCIS (Netherlands)

    Derks, P. W. J.; Versteeg, G. F.

    2009-01-01

    In the present work the absorption of carbon dioxide into aqueous ammonia solutions has been studied in a stirred cell reactor, at low temperatures and ammonia concentrations ranging from 0.1 to about 7 kmol m-3. The absorption experiments were carried out at conditions where the so-called pseudo

  1. Ammonia treated Mo/AC catalysts for CO hydrogenation with ...

    Indian Academy of Sciences (India)

    A series of ammonia treated Mo/Activated Carbon (AC) catalysts were synthesized by wet impregnation method by nominal incorporation of 5, 10 and 15 wt% of molybdenum. The calcined catalysts (500◦C, 4 h, N₂ flow) were subjected to a stepwise ammonia treatment at temperatures from 25 up to 700◦C. This work ...

  2. Ammonia loss, ammonium and nitrate accumulation from mixing ...

    African Journals Online (AJOL)

    Ammonia loss from urea significantly hinders efficient use of urea in agriculture. In order to reduce ammonia loss and, at the same time, improve beneficial accumulation of soil exchangeable ammonium and nitrate for efficient utilization by plants, this laboratory study was conducted to determine the effect of mixing urea with ...

  3. Consumption of ammonia-nitrogen by aob in immobilized batch ...

    African Journals Online (AJOL)

    This study investigated the performance of bio-filters immobilized nitrifying bacteria, ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB). In particular, it was to assess the consumption of ammonia-nitrogen (NH3-N) by the AOB. The experiment was conducted in a one litre reactor.The reactor consisted of ...

  4. Disinfection kinetics of pathogens in physicochemical sludge treated with ammonia.

    Science.gov (United States)

    Mendez, J M; Jimenez, B; Maya, C

    2004-01-01

    Ammonia is a disinfectant which can diffuse through the membrane of highly resistant structures like helminth ova. Thus, it can be considered an alternative disinfectant of wastewater sludge with high pathogenic content. In this study, the kinetic parameters of the Hom model were used to describe the inactivation with ammonia of faecal coliforms, Salmonella spp. and viable helminth ova. These were obtained in processes considering the addition of ammonia alone as well as for ammonia combined with an increase in temperature. The sludge was sampled from a municipal wastewater treatment plant using an APT (Advanced Primary Treatment) or CEP (Chemical Enhanced Primary) process. With 20% w/w of ammonia, 7 logs of faecal coliforms, 6 logs of Salmonella spp., and 83% of viable helminth ova were reduced in 2 hours contact time. To eliminate 100% of the helminth ova from samples having 88-132 ova/g TS it was needed to combine 20% of ammonia with 50 degrees C. The analysis of parameters k, n and m indicate higher resistance to inactivation of helminth ova compared to bacteria and a better performance of the ammonia process than lime stabilization to inactivate microorganisms. In addition, ammonia increased the agricultural value of the biosolids produced.

  5. Lignin solubilisation and gentle fractionation in liquid ammonia

    NARCIS (Netherlands)

    Strassberger, Z.; Prinsen, P.; Klis, van der F.; Es, van D.S.; Tanase, S.; Rothenberg, G.

    2015-01-01

    We present a simple method for solubilising lignin using liquid ammonia. Unlike water, which requires harsh conditions, ammonia can solubilise technical lignins, in particular kraft lignin. A commercial pine wood Kraft lignin (Indulin AT) was solubilized instantaneously at room temperature and 7–11

  6. Thermodynamic Model for the Ammonia-Water System

    DEFF Research Database (Denmark)

    Thomsen, Kaj; Rasmussen, Peter

    2000-01-01

    The ammonia-water system is described by the Extended UNIQUAC model, which is an electrolyte model, formed by combining the original UNIQUAC model, the Debye-Hückel law and the Soave-Redlich-Kwong equation of state. The model is limited to temperatures below the critical temperature of ammonia. V...

  7. Treatment of ammonia in liquid hospital waste using activated carbon

    Science.gov (United States)

    Riyanto, Hayati, Lena

    2017-12-01

    In this research study of the treatment of ammonia in liquid hospitals waste using activated carbon. This study aims to the effect of activated carbon weight and precipitation time to the treatment of ammonia in liquid hospitals waste. Hospital liquid waste has been taken from Jogja International Hospital (JIH) Yogyakarta, Indonesia. Hospital liquid waste 100 mL is mixed with activated carbon with the varied weight that is 15, 30 and 60 g. After added with activated carbon then stirred with a magnetic stirrer for 15 minutes and a precipitation time of 0.5, 1.0 and 2.0 hours. The next step is the filtrate analyzed ammonia concentrations before and after treatment using UV-Vis Spectrophotometer. The results showed that activated carbon can reduce ammonia concentration in hospital liquid waste. The amount of the active carbon and the time of stirring, the greater the ammonia concentration decreases in hospital liquid waste. The best condition for the decrease of the ammonia concentration was obtained with active carbon and precipitation time is 60 g and 1.0 hours, respectively with ammonia decrease of 95.93%. The conclusion is that activated carbon can reduce ammonia concentration in hospital liquid waste.

  8. Alkaline Ammonia Electrolysis on Electrodeposited Platinum for Controllable Hydrogen Production.

    Science.gov (United States)

    Gwak, Jieun; Choun, Myounghoon; Lee, Jaeyoung

    2016-02-19

    Ammonia is beginning to attract a great deal of attention as an alternative energy source carrier, because clean hydrogen can be produced through electrolytic processes without the emission of COx . In this study, we deposited various shapes of Pt catalysts under potentiostatic mode; the electrocatalytic oxidation behavior of ammonia using these catalysts was studied in alkaline media. The electrodeposited Pt was characterized by both qualitative and quantitative analysis. To discover the optimal structure and the effect of ammonia concentration, the bulk pH value, reaction temperature, and applied current of ammonia oxidation were investigated using potential sweep and galvanostatic methods. Finally, ammonia electrolysis was conducted using a zero-gap cell, producing highly pure hydrogen with an energy efficiency over 80 %. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Ammonia effect on hydrogenotrophic methanogens and syntrophic acetate oxidizing bacteria

    DEFF Research Database (Denmark)

    Wang, Han; Fotidis, Ioannis; Angelidaki, Irini

    2015-01-01

    Ammonia-rich substrates can cause inhibition on anaerobic digestion process. Syntrophic acetate oxidizing bacteria (SAOB) and hydrogenotrophic methanogens are important for the ammonia inhibitory mechanism on anaerobic digestion. The roles and interactions of SAOB and hydrogenotrophic methanogens...... to ammonia inhibition effect are still unclear. The aim of the current study was to determine the ammonia toxicity levels of various pure strains of SAOB and hydrogenotrophic methanogens. Moreover, ammonia toxicity on the syntrophic cultivated strains of SAOB and hydrogenotrophic methanogens was tested. Thus......, four hydrogenotrophic methanogens (i.e. Methanoculleus bourgensis, Methanobacterium congolense, Methanoculleus thermophilus and Methanothermobacter thermautotrophicus), two SAOB (i.e. Tepidanaerobacter acetatoxydans and Thermacetogenium phaeum) and their syntrophic cultivation, were assessed under 0...

  10. Safety assessment of ammonia as a transport fuel

    DEFF Research Database (Denmark)

    Duijm, N.J.; Markert, Frank; Paulsen, Jette Lundtang

    2005-01-01

    of transport of ammonia to the refuelling stations and safety of the activities at the refuelling station (unloading and refuelling). Comparisons are made between the safety of using ammonia and the safety of otherexisting or alternative fuels. The conclusion is that the hazards in relation to ammonia need...... to be controlled by a combination of technical and regulatory measures. The most important requirements are: - Advanced safety systems in the vehicle -Additional technical measures and regulations are required to avoid releases in maintenance workshops and unauthorised maintenance on the fuel system. - Road...... transport of ammonia to refuelling stations in refrigerated form - Sufficient safety zonesbetween refuelling stations and residential or otherwise public areas. When these measures are applied, the use of ammonia as a transport fuel wouldn’t cause more risks than currently used fuels (using current practice)....

  11. Optimization of biomethanation focusing on high ammonia loaded processes

    DEFF Research Database (Denmark)

    Wang, Han

    The toxicity effect of high ammonia is one of the most common problems, which cause imbalance and low biogas production rate in biogas plants. When protein-rich substrates (e.g. pig manure and mink manure, food waste, etc.) are used in biogas plants, lead to suboptimal utilization of the biogas...... potential and unstable biogas process. However, up to now, the solutions for alleviating ammonia toxicity effect have been proven either too expensive or time consuming for the full-scale biogas plants. Thus, sustainable and practical solutions to overcome the problem of ammonia inhibition efficiently...... and subsequently been converted together with carbon dioxide to methane by hydrogenotrophic methanogens, could potentially be more tolerant to ammonia toxicity. Therefore, the effect of different ammonia levels on this hydrogen assisted biogas upgrading process under different hydrogen partial pressure (0, 0.25, 0...

  12. Methods of ammonia removal in anaerobic digestion: a review.

    Science.gov (United States)

    Krakat, Niclas; Demirel, Burak; Anjum, Reshma; Dietz, Donna

    2017-10-01

    The anaerobic digestion of substrates with high ammonia content has always been a bottleneck in the methanisation process of biomasses. Since microbial communities in anaerobic digesters are sensitive to free ammonia at certain conditions, the digestion of nitrogen-rich substrates such as livestock wastes may result in inhibition/toxicity eventually leading to process failures, unless appropriate engineering precautions are taken. There are many different options reported in literature to remove ammonia from anaerobic digesters to achieve a safe and stable process so that along with high methane yields, a good quality of effluents can also be obtained. Conventional techniques to remove ammonia include physical/chemical methods, immobilization and adaptation of microorganisms, while novel methods include ultrasonication, microwave, hollow fiber membranes and microbial fuel cell applications. This paper discusses conventional and novel methods of ammonia removal from anaerobic digesters using nitrogen-rich substrates, with particular focus on recent literature available about this topic.

  13. Modelling of Cavitation of Wash-Out Water, Ammonia Water, Ammonia Water with Increased Content Ammonia and Hydrogen Sulphide, Tar Condensate

    Directory of Open Access Journals (Sweden)

    Josef DOBEŠ

    2013-12-01

    Full Text Available The aim is to design and implement a procedure of numerical modelling of cavitation of working mixtures: wash-out water, ammonia water, ammonia water with an increased content of hydrogen sulphide and ammonia, tar condensate. The numeric modelling is designed in the program Ansys Fluent using Schnerr-Sauer cavitation model. The issue of these liquids modelling can be solved by the cavitation simulation of water admixtures. Working fluids contain the following main ingredients: water, ammonia, carbon dioxide and hydrogen sulphide. Subsequently, a comparison of the amount of water vapor (reference liquid and given fluid vapor is executed. The Schnerr-Sauer model is chosen because of good results in previous simulations for water cavitation. As a geometry is selected Laval nozzle. Modelled liquid mixtures are used in the petrochemical industry, as a filling for fluid circuits where cavitation may occur and therefore the research is needed.

  14. [Properties of alkaline pectate lyase from recombinant strain E. coli JM109 (pHsh PL)].

    Science.gov (United States)

    Zhuge, Bin; Du, Guocheng; Zhuge, Jian; Chen, Jian

    2008-01-01

    Alkaline pectate lyase (PL) from recombinant strain E. coli JM109 (pHsh PL) was purified by a three-step process including (NH4)2SO4 precipitation followed by dialysis and chromatography. The purified enzyme appeared homologous on SDS-PAGE. The specific activity of the purified enzyme reached 1079 U/mg. The optimal pH and temperature were in the ranges of pH 9.0 to 10.0 and 50 degrees C to 66 degrees C. The enzyme was preferable in optimal pH range in enzymatic retting of flax. Enzyme activity slightly increased in the presence of Mg2+ ion, whereas decreased in the presence of other ions, especially Fe2+. The K(m) of the purified enzyme for polygalacturonic acid was 20.93 mg/L, the V(max) for polygalacturonic acid hydrolysis was 105.3 micromol of unsaturated products per min and Ea was 21.74 kJ/mol. The results of the decay constant (k(d)) analysis on condition of PL bonding polygalacturonic acid (k(d) = 0.02 min(-1)) and PL without polygalacturonic acid (k(d) = 0.0342 min(-1)) showed the substrate was helpful to decrease thermal inactivation of PL. The products (unsaturated oligomers) from polygalacturonic acid degraded by PL were analyzed by electrospray ionization mass spectrometry(ESI-MS). The following data were obtained: ESI-MS m/z, 350.82 (unsaturated bigalacturonic acid, uG2), 527.04 (unsaturated trigalacturonic acid, uG3). However, m/z 175 (unsaturated galacturonic acid, uGI) was not found. These results indicate that the final PGA degradation products was a mixture of unsaturated oligo-galacturonides including uG3 and uG2 except for uG1. It suggests that the recombinant PL cannot degrade uG3 and uG2.

  15. Molecular identification and pectate lyase production by Bacillus strains involved in cocoa fermentation.

    Science.gov (United States)

    Ouattara, Honoré G; Reverchon, Sylvie; Niamke, Sébastien L; Nasser, William

    2011-02-01

    We have previously reported the implication of Bacillus in the production of pectinolytic enzymes during cocoa fermentation. The objective of this work was to identify the Bacillus strains isolated from cocoa fermentation and study their ability to produce pectate lyase (PL) in various growth conditions. Ninety-eight strains were analyzed by Amplified Ribosomal DNA Restriction Analysis (ARDRA). Four different banding patterns were obtained leading to the clustering of the bacterial isolates into 4 distinct ARDRA groups. A subset of representative isolates for each group was identified by 16S rRNA gene partial sequencing. Six species were identified: Bacillus subtilis, Bacillus pumilus, Bacillus sphaericus, Bacillus cereus, Bacillus thuringiensis, together with Bacillus fusiformis which was isolated for the first time from cocoa fermentation. The best PL producers, yielding at least 9 U/mg of bacterial dry weight, belonged to B. fusiformis, B. subtilis, and B. pumilus species while those belonging to B. sphaericus, B. cereus and B. thuringiensis generally showed a low level of activity. Two kinds of PL were produced, as revealed by isoelectrofocusing: one with a pI of 9.8 produced by B. subtilis and B. fusiformis, the other with a pI of 10.5 was produced by B. pumilus. Strains yielded about 2 fold more PL in a pectic compound medium than in glucose medium and maximum enzyme production occurred in the late stationary bacterial growth phase. Together all these results indicate that PL production in the bacilli studied is modulated by the growth phase and by the carbon source present in the medium. Copyright © 2010 Elsevier Ltd. All rights reserved.

  16. Characterization of a bifunctional glyoxylate cycle enzyme, malate synthase/isocitrate lyase, of Euglena gracilis.

    Science.gov (United States)

    Nakazawa, Masami; Nishimura, Masaaki; Inoue, Kengo; Ueda, Mitsuhiro; Inui, Hiroshi; Nakano, Yoshihisa; Miyatake, Kazutaka

    2011-01-01

    The glyoxylate cycle is a modified form of the tricarboxylic acid cycle, which enables organisms to synthesize carbohydrates from C2 compounds. In the protozoan Euglena gracilis, the key enzyme activities of the glyoxylate cycle, isocitrate lyase (ICL) and malate synthase (MS), are conferred by a single bifunctional protein named glyoxylate cycle enzyme (Euglena gracilis glyoxylate cycle enzyme [EgGCE]). We analyzed the enzymatic properties of recombinant EgGCE to determine the functions of its different domains. The 62-kDa N-terminal domain of EgGCE was sufficient to provide the MS activity as expected from an analysis of the deduced amino acid sequence. In contrast, expression of the 67-kDa C-terminal domain of EgGCE failed to yield ICL activity even though this domain was structurally similar to ICL family enzymes. Analyses of truncation mutants suggested that the N-terminal residues of EgGCE are critical for both the ICL and MS activities. The ICL activity of EgGCE increased in the presence of micro-molar concentrations of acetyl-coenzyme A (CoA). Acetyl-CoA also increased the activity in a mutant type EgGCE with a mutation at the acetyl-CoA binding site in the MS domain of EgGCE. This suggests that acetyl-CoA regulates the ICL reaction by binding to a site other than the catalytic center of the MS reaction. © 2011 The Author(s). Journal of Eukaryotic Microbiology© 2011 International Society of Protistologists.

  17. Structural and kinetic analysis of Schistosoma mansoni Adenylosuccinate Lyase (SmADSL).

    Science.gov (United States)

    Romanello, Larissa; Serrão, Vitor Hugo Balasco; Torini, Juliana Roberta; Bird, Louise E; Nettleship, Joanne E; Rada, Heather; Reddivari, Yamini; Owens, Ray J; DeMarco, Ricardo; Brandão-Neto, José; Pereira, Humberto D'Muniz

    2017-06-01

    Schistosoma mansoni is the parasite responsible for schistosomiasis, a disease that affects about 218 million people worldwide. Currently, both direct treatment and disease control initiatives rely on chemotherapy using a single drug, praziquantel. Concerns over the possibility of resistance developing to praziquantel, have stimulated efforts to develop new drugs for the treatment of schistosomiasis. Schistosomes do not have the de novo purine biosynthetic pathway, and instead depend entirely on the purine salvage pathway to supply its need for purines. The purine salvage pathway has been reported as a potential target for developing new drugs against schistosomiasis. Adenylosuccinate lyase (SmADSL) is an enzyme in this pathway, which cleaves adenylosuccinate (ADS) into adenosine 5'-monophosphate (AMP) and fumarate. SmADSL kinetic characterization was performed by isothermal titration calorimetry (ITC) using both ADS and SAICAR as substrates. Structures of SmADSL in Apo form and in complex with AMP were elucidated by x-ray crystallography revealing a highly conserved tetrameric structure required for their function since the active sites are formed from residues of three different subunits. The active sites are also highly conserved between species and it is difficult to identify a potent species-specific inhibitor for the development of new therapeutic agents. In contrast, several mutagenesis studies have demonstrated the importance of dimeric interface residues in the stability of the quaternary structure of the enzyme. The lower conservation of these residues between SmADSL and human ADSL could be used to lead the development of anti-schistosomiasis drugs based on disruption of subunit interfaces. These structures and kinetics data add another layer of information to Schistosoma mansoni purine salvage pathway. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Pyruvate formate lyase acts as a formate supplier for metabolic processes during anaerobiosis in Staphylococcus aureus.

    Science.gov (United States)

    Leibig, Martina; Liebeke, Manuel; Mader, Diana; Lalk, Michael; Peschel, Andreas; Götz, Friedrich

    2011-02-01

    Previous studies demonstrated an upregulation of pyruvate formate lyase (Pfl) and NAD-dependent formate dehydrogenase (Fdh) in Staphylococcus aureus biofilms. To investigate their physiological role, we constructed fdh and pfl deletion mutants (Δfdh and Δpfl). Although formate dehydrogenase activity in the fdh mutant was lost, it showed little phenotypic alterations under oxygen-limited conditions. In contrast, the pfl mutant displayed pleiotropic effects and revealed the importance of formate production for anabolic metabolism. In the pfl mutant, no formate was produced, glucose consumption was delayed, and ethanol production was decreased, whereas acetate and lactate production were unaffected. All metabolic alterations could be restored by addition of formate or complementation of the Δpfl mutant. In compensation reactions, serine and threonine were consumed better by the Δpfl mutant than by the wild type, suggesting that their catabolism contributes to the refilling of formyl-tetrahydrofolate, which acts as a donor of formyl groups in, e.g., purine and protein biosynthesis. This notion was supported by reduced production of formylated peptides by the Δpfl mutant compared to that of the parental strain, as demonstrated by weaker formyl-peptide receptor 1 (FPR1)-mediated activation of leukocytes with the mutant. FPR1 stimulation could also be restored either by addition of formate or by complementation of the mutation. Furthermore, arginine consumption and arc operon transcription were increased in the Δpfl mutant. Unlike what occurred with the investigated anaerobic conditions, a biofilm is distinguished by nutrient, oxygen, and pH gradients, and we thus assume that Pfl plays a significant role in the anaerobic layer of a biofilm. Fdh might be critical in (micro)aerobic layers, as formate oxidation is correlated with the generation of NADH/H(+), whose regeneration requires respiration.

  19. Partial deficiency of sphingosine-1-phosphate lyase confers protection in experimental autoimmune encephalomyelitis.

    Directory of Open Access Journals (Sweden)

    Andreas Billich

    Full Text Available BACKGROUND: Sphingosine-1-phosphate (S1P regulates the egress of T cells from lymphoid organs; levels of S1P in the tissues are controlled by S1P lyase (Sgpl1. Hence, Sgpl1 offers a target to block T cell-dependent inflammatory processes. However, the involvement of Sgpl1 in models of disease has not been fully elucidated yet, since Sgpl1 KO mice have a short life-span. METHODOLOGY: We generated inducible Sgpl1 KO mice featuring partial reduction of Sgpl1 activity and analyzed them with respect to sphingolipid levels, T-cell distribution, and response in models of inflammation. PRINCIPAL FINDINGS: The partially Sgpl1 deficient mice are viable but feature profound reduction of peripheral T cells, similar to the constitutive KO mice. While thymic T cell development in these mice appears normal, mature T cells are retained in thymus and lymph nodes, leading to reduced T cell numbers in spleen and blood, with a skewing towards increased proportions of memory T cells and T regulatory cells. The therapeutic relevance of Sgpl1 is demonstrated by the fact that the inducible KO mice are protected in experimental autoimmune encephalomyelitis (EAE. T cell immigration into the CNS was found to be profoundly reduced. Since S1P levels in the brain of the animals are unchanged, we conclude that protection in EAE is due to the peripheral effect on T cells, leading to reduced CNS immigration, rather than on local effects in the CNS. SIGNIFICANCE: The data suggest Sgpl1 as a novel therapeutic target for the treatment of multiple sclerosis.

  20. Simultaneous determination of the lipoxygenase and hydroperxide lyase specificity in olive fruit pulp

    Directory of Open Access Journals (Sweden)

    Salas, Joaquín J.

    2000-06-01

    Full Text Available Olive pulp lipoxygenase regiospecificity and hydroperoxide lyase substrate specificity are important parameters in order to justify the volatile composition of olive oil. A new radiolabelling method to determine simultaneously these properties using only thin layer chromatography steps is described in the present work. The method involves incubation of an enzyme preparation from olive pulp with radiolabelled linoleate, followed by the fractionation of the resulting lipid products, previously treated with 2,4-dinitrophenyl hydrazine, on thin layer chromatography plates coated with polyethylenglycol 400. The results obtained are in agreement with previous studies carried out by other methods.La regioespecificidad de la lipoxigenasa y la especificidad del sustrato hidroperóxido liasa de pulpa de aceituna son parámetros importantes en la justificación de la composición en volátiles del aceite de oliva. En este trabajo se describe un nuevo método de marcaje radioactivo para determinar simultáneamente estas propiedades, usando solo etapas de cromatografía en capa fina. El método implica la incubación de una preparación enzimática de pulpa de aceituna con linoleato marcado, seguido del fraccionamiento de los productos lipídicos resultantes, previamente tratados con 2,4-dinitrofenil hidrazina, sobre placas de cromatografía en capa fina soportadas con polietilenglicol 400. Los resultados obtenidos están de acuerdo con estudios previos llevados a cabo con otros métodos.

  1. Ammonia emissions in agriculture: Proceedings of the First international ammonia conference 19-21 March 2007, Ede, The Netherlands

    NARCIS (Netherlands)

    Monteny, G.J.; Hartung, J.

    2007-01-01

    Ammonia emissions is an important topic in many countries with animal production, since it contributes to environmental and health problems. Strategies and measures to reduce ammonia emission are getting increasing attention in national and international legislation. The focus of this publication is

  2. Arterial ammonia with Blood Ammonia Checker II and with indophenol reaction to assess presence of hepatic encephalopathy

    NARCIS (Netherlands)

    Huizenga, [No Value; vanDam, GM; Gips, CH

    1996-01-01

    Hepatic encephalopathy (HE) is associated with elevated arterial ammonia levels. The relationship is variable, in part due to ammonia methodology. One method, based on the indophenol reaction (IPh), is interfered with a number of amino acids including all aromatic amino acids. We have determined

  3. Doppler spectroscopy of hydrogen Balmer lines in a hollow cathode glow discharge in ammonia and argon-ammonia mixture

    International Nuclear Information System (INIS)

    Sisovic, N. M.; Konjevic, N.

    2008-01-01

    The results of Doppler spectroscopy of hydrogen Balmer lines from a stainless steel (SS) and copper (Cu) hollow cathode (HC) glow discharge in ammonia and argon-ammonia mixture are reported. The experimental profiles in ammonia discharge are fitted well by superposing three Gaussian profiles. The half widths, in energy units, of narrow and medium Gaussians are in the ranges 0.3-0.4 eV and 3-4 eV, respectively, for both hollow cathodes what is expected on the basis of earlier electron beam→NH 3 experiments. The half widths of the largest Gaussian in ammonia are 46 and 55 eV for SS and Cu HC, respectively. In argon-ammonia discharge, three Gaussians are also required to fit experimental profiles. While half widths of narrow and medium Gaussians are similar to those in ammonia, the half widths of the largest Gaussians are 35 and 42 eV for SS and Cu HC, respectively. The half widths of the largest Gaussians in ammonia and in argon-ammonia mixture indicate the presence of excessive Doppler broadening.

  4. New Mechanistic Insight from Substrate- and Product-Bound Structures of the Metal-Dependent Dimethylsulfoniopropionate Lyase DddQ.

    Science.gov (United States)

    Brummett, Adam E; Dey, Mishtu

    2016-11-08

    The marine microbial catabolism of dimethylsulfoniopropionate (DMSP) by the lyase pathway liberates ∼300 million tons of dimethyl sulfide (DMS) per year, which plays a major role in the biogeochemical cycling of sulfur. Recent biochemical and structural studies of some DMSP lyases, including DddQ, reveal the importance of divalent transition metal ions in assisting DMSP cleavage. While DddQ is believed to be zinc-dependent primarily on the basis of structural studies, excess zinc inhibits the enzyme. We examine the importance of iron in regulating the DMSP β-elimination reaction catalyzed by DddQ as our as-isolated purple-colored enzyme possesses ∼0.5 Fe/subunit. The UV-visible spectrum exhibited a feature at 550 nm, consistent with a tyrosinate-Fe(III) ligand-to-metal charge transfer transition. Incubation of as-isolated DddQ with added iron increases the intensity of the 550 nm peak, whereas addition of dithionite causes a bleaching as Fe(III) is reduced. Both the Fe(III) oxidized and Fe(II) reduced species are active, with similar k cat values and 2-fold differences in their K m values for DMSP. The slow turnover of Fe(III)-bound DddQ allowed us to capture a substrate-bound form of the enzyme. Our DMSP-Fe(III)-DddQ structure reveals conformational changes associated with substrate binding and shows that DMSP is positioned optimally to bind iron and is in the proximity of Tyr 120 that acts as a Lewis base to initiate catalysis. The structures of Tris-, DMSP-, and acrylate-bound forms of Fe(III)-DddQ reported here illustrate various states of the enzyme along the reaction pathway. These results provide new insights into DMSP lyase catalysis and have broader significance for understanding the mechanism of oceanic DMS production.

  5. Molecular cloning and characterization of an Erwinia carotovora subsp. carotovora pectin lyase gene that responds to DNA-damaging agents.

    OpenAIRE

    McEvoy, J L; Murata, H; Chatterjee, A K

    1990-01-01

    recA-mediated production of pectin lyase (PNL) and the bacteriocin carotovoricin occurs in Erwinia carotovora subsp. carotovora 71 when this organism is subjected to agents that damage or inhibit the synthesis of DNA. The structural gene pnlA was isolated from a strain 71 cosmid gene library following mobilization of the cosmids into a moderate PNL producer, strain 193. The cosmid complemented pnl::Tn5 but not ctv::Tn5 mutations. A constitutive level of PNL activity was detected in RecA+ and ...

  6. Insight into the role of substrate-binding residues in conferring substrate specificity for the multifunctional polysaccharide lyase Smlt1473.

    Science.gov (United States)

    MacDonald, Logan C; Berger, Bryan W

    2014-06-27

    Anionic polysaccharides are of growing interest in the biotechnology industry due to their potential pharmaceutical applications in drug delivery and wound treatment. Chemical composition and polymer length strongly influence the physical and biological properties of the polysaccharide and thus its potential industrial and medical applications. One promising approach to determining monomer composition and controlling the degree of polymerization involves the use of polysaccharide lyases, which catalyze the depolymerization of anionic polysaccharides via a β-elimination mechanism. Utilization of these enzymes for the production of custom-made oligosaccharides requires a high degree of control over substrate specificity. Previously, we characterized a polysaccharide lyase (Smlt1473) from Stenotrophomonas maltophilia k279a, which exhibited significant activity against hyaluronan (HA), poly-β-d-glucuronic acid (poly-GlcUA), and poly-β-d-mannuronic acid (poly-ManA) in a pH-regulated manner. Here, we utilize a sequence structure guided approach based on a homology model of Smlt1473 to identify nine putative substrate-binding residues and examine their effect on substrate specificity via site-directed mutagenesis. Interestingly, single point mutations H221F and R312L resulted in increased activity and specificity toward poly-ManA and poly-GlcUA, respectively. Furthermore, a W171A mutant nearly eliminated HA activity, while increasing poly-ManA and poly-GlcUA activity by at least 35%. The effect of these mutations was analyzed by comparison with the high resolution structure of Sphingomonas sp. A1-III alginate lyase in complex with poly-ManA tetrasaccharide and by taking into account the structural differences between HA, poly-GlcUA, and poly-ManA. Overall, our results demonstrate that even minor changes in active site architecture have a significant effect on the substrate specificity of Smlt1473, whose structural plasticity could be applied to the design of highly

  7. Functional pyruvate formate lyase pathway expressed with two different electron donors in Saccharomyces cerevisiae at aerobic growth

    DEFF Research Database (Denmark)

    Zhang, Yiming; Dai, Zongjie; Krivoruchko, Anastasia

    2015-01-01

    Pyruvate formate lyase (PFL) is characterized as an enzyme functional at anaerobic conditions, since the radical in the enzyme's active form is sensitive to oxygen. In this study, PFL and its activating enzyme from Escherichia coli were expressed in a Saccharomyces cerevisiae strain lacking...... of either of these electron donors had a positive effect on growth under aerobic conditions, indicating increased activity of PFL. The positive effect on growth was manifested as a higher final biomass concentration and a significant increase in transcription of formate dehydrogenases. Among the two...

  8. Functional implications of the beta-helical protein fold: differences in chemical and thermal stabilities of Erwinia chrysanthemi EC16 pectate lyases B, C, and E.

    Science.gov (United States)

    Hurlbert, J C; Preston, J F

    2000-09-15

    Colonization of plant tissue by the phytopathogen Erwinia chrysanthemi EC16 is aided by the activities of the pectate lyase isozymes (PLs), which depolymerize the polygalacturonic acid component (PGA) of plant cell walls. The bacterium secretes four pectate lyases (PLa, PLb, PLc, and PLe), two of which, PLc and PLe, have been shown to fold into a similar domain motif, the beta-helix. To understand the rationale behind the evolution and retention of these isoforms, the susceptibilities of pectate lyases B, C, and E to chemical and thermal denaturation and the resulting enzymatic inactivation were examined. With guanidine hydrochloride used as a denaturant, all three pectate lyases denatured with transition midpoint guanidine hydrochloride concentrations (Cm) of 1.3, 1.1, and 1.8 M for PLb, PLc, and PLe, respectively. Lyase activity decreased in direct response to loss of secondary structure in all enzymes. Pectate lyases B and C demonstrated increased enzymatic activity at temperatures above 30 degrees C, with maximal activity observed at 40 degrees C for PLb and 35 degrees C for PLc. Transition midpoints (Tm) as measured by circular dichroism were at 46.9 degrees C for PLb and 44.3 degrees C for PLc, indicating detectable conformational changes accompanying thermal inactivation. Decreased enzymatic activity of PLe was observed at all temperatures above 30 degrees C, and the enzyme was found to possess a Tm at 38.9 degrees C. The data demonstrate structural differences among these enzymes that may be the basis for different enzymatic efficiencies under the potential array of environmental conditions experienced by the bacterium. These differences, in turn, may play a part in the retention of these isozymes as virulence factors, allowing the successful colonization of susceptible plant hosts.

  9. Ammonia-Free NOx Control System

    Energy Technology Data Exchange (ETDEWEB)

    Song Wu; Zhen Fan; Andrew H. Seltzer; Richard G. Herman

    2004-09-30

    Research is being conducted under United States Department of Energy (DOE) Contract DEFC26-03NT41865 to develop a new technology to achieve very low levels of NOx emissions from pulverized coal fired boiler systems by employing a novel system level integration between the PC combustion process and the catalytic NOx reduction with CO present in the combustion flue gas. The combustor design and operating conditions will be optimized to achieve atypical flue gas conditions. This approach will not only suppress NOx generation during combustion but also further reduce NOx over a downstream catalytic reactor that does not require addition of an external reductant, such as ammonia. This report describes the work performed during the July 1 to September 30, 2004 time period.

  10. Ammonia-Free NOx Control System

    Energy Technology Data Exchange (ETDEWEB)

    Song Wu; Zhen Fan; Andrew H. Seltzer

    2005-09-30

    Research is being conducted under United States Department of Energy (DOE) Contract DEFC26-03NT41865 to develop a new technology to achieve very low levels of NOx emissions from pulverized coal fired boiler systems by employing a novel system level integration between the PC combustion process and the catalytic NOx reduction with CO present in the combustion flue gas. The combustor design and operating conditions will be optimized to achieve atypical flue gas conditions. This approach will not only suppress NOx generation during combustion but also further reduce NOx over a downstream catalytic reactor that does not require addition of an external reductant, such as ammonia. This report describes the work performed during the July 1 to September 30, 2005 time period.

  11. Ammonia-Free NOx Control System

    Energy Technology Data Exchange (ETDEWEB)

    Song Wu; Zhen Fan; Richard G. Herman

    2004-12-31

    Research is being conducted under United States Department of Energy (DOE) Contract DEFC26-03NT41865 to develop a new technology to achieve very low levels of NOx emissions from pulverized coal fired boiler systems by employing a novel system level integration between the PC combustion process and the catalytic NOx reduction with CO present in the combustion flue gas. The combustor design and operating conditions will be optimized to achieve atypical flue gas conditions. This approach will not only suppress NOx generation during combustion but also further reduce NOx over a downstream catalytic reactor that does not require addition of an external reductant, such as ammonia. This report describes the work performed during the October 1 to December 30, 2004 time period.

  12. Towards an ammonia-mediated hydrogen economy?

    DEFF Research Database (Denmark)

    Christensen, Claus H.; Johannessen, Tue; Sørensen, Rasmus Zink

    2006-01-01

    . Recently, metal ammine salts were proposed as safe, reversible. high-density and low-cost hydrogen carriers. Here, we discuss how this development could provide a platform for using ammonia as a fuel for the hydrogen economy, We do that by comparing various possible hydrogen carriers with respect to energy......Materialization of a hydrogen economy could provide a solution to significant global challenges, In particular. the possibility of improving the efficiency and simultaneously minimizing the environmental impact of energy conversion processes, together with the opportunity to reduce the dependency...... of fossil fuels, are main drivers for the currently increasing research and development efforts. However. significant technological breakthroughs are necessary for making a hydrogen economy feasible. In particular, it is necessary to develop appropriate hydrogen storage and transportation technologies...

  13. Study of ammonia synthesis over uranium catalysts

    International Nuclear Information System (INIS)

    Spitsyn, V.I.; Erofeev, B.V.; Mikhajlenko, I.E.; Gorelkin, I.I.; Ivanov, L.S.

    1980-01-01

    The effect of induced radiactivity and chemical composition of uranium catalysts on their catalytic activity in the ammonia synthesis reaction has been studied. The catalyst samples comprise pieces of metal uranium and chip irradiated in nuclear reactor by the 4.3x10 16 n/cm 2 integral flux of slow neutrons. Studies of catalytic activity was carried out at 1 atm and 340-510 deg C when stoichiometric nitrogen-hydrogen mixture passed through the following installation. At different temperatures uranium nitrides of different composition are shown to be formed. Uranium nitrides with the composition close to UN 2 are the samples with the highest catalYtic activity. The reduction of catalytic activity of uranium catalysts with the increased temperature of their formation above 400 deg C is explained by low catalytic activity of forming UNsub(1.7) in comparison with UN 2 . Catalytic properties of irradiated and nonirradiated samples do not differ from one another

  14. Syntrophy of aerobic and anaerobic ammonia oxidisers.

    Science.gov (United States)

    Wett, B; Hell, M; Nyhuis, G; Puempel, T; Takacs, I; Murthy, S

    2010-01-01

    Deammonification is known as an efficient and resource saving sidestream process option to remove the nitrogen load from sludge liquors. The transfer of the intermediate product nitrite between both syntrophic groups of organisms - aerobic and anaerobic ammonia oxidizers (AOB) - appears very sensitive to process conditions such as temperature, dissolved oxygen (DO) and operating nitrite level. Growth kinetics for aerobic and anaerobic AOBs differ by one order of magnitude and require an adequate selection of sludge retention time. This paper provides measurement- and model-based results on how selected sludge wasting impacts population dynamics in a suspended growth deammonification system. Anammox enrichment up to a doubled portion in mixed liquor solids can substantially improve process stability in difficult conditions. A case-study on low temperature operations outlines two possible strategies to balance syntrophic consumption of ammonium and nitrite.

  15. Nitrogen-13-labeled ammonia for myocardial imaging

    Energy Technology Data Exchange (ETDEWEB)

    Walsh, W.F.; Fill, H.R.; Harper, P.V.

    1977-01-01

    Cyclotron-produced nitrogen-13 (half-life 10 min), as labeled ammonia (/sup 13/NH/sub 4//sup +/), has been evaluated as a myocardial perfusion imaging agent. The regional myocardial uptake of /sup 13/NH/sub 4//sup +/ has been shown to be proportional to regional tissue perfusion in animal studies. Intravenously administered /sup 13/NH/sub 4//sup +/ is rapidly cleared from the circulation, being extracted by the liver (15 percent), lungs, myocardium (2 percent--4 percent), brain, kidney, and bladder. Myocardial ammonia is metabolized mainly to glutamine via the glutamine synthetase pathway. Pulmonary uptake is substantial, but usually transient, except in smokers where clearance may be delayed. The positron annihilation irradiation (511 keV) of /sup 13/N may be imaged with a scintillation camera, using either a specially designed tungsten collimator or a pinhole collimator. After early technical problems with collimation and the production method of /sup 13/NH/sub 4//sup +/ were overcome, reproducible high quality myocardial images were consistently obtained. The normal myocardial image was established to be of a homogeneous ''doughnut'' configuration. Imaging studies performed in patients with varying manifestations of ischemic and valvular heart disease showed a high incidence of localized perfusion defects, especially in patients with acute myocardial infarction. Sequential studies at short intervals in patients with acute infarction showed correlation between alterations in regional perfusion and the clinical course of the patient. It is concluded that myocardial imaging with /sup 13/NH/sub 4//sup +/ and a scintillation camera provides a valid and noninvasive means of assessing regional myocardial perfusion. This method is especially suitable for sequential studies of acute cardiac patients at short intervals. Coincidence imaging of the 511 keV annihilation irradiation provides a tomographic and potentially quantitative assessment of the

  16. Nitrogen metabolism and kinetics of ammonia-oxidizing archaea.

    Science.gov (United States)

    Martens-Habbena, Willm; Stahl, David A

    2011-01-01

    The discovery of ammonia-oxidizing mesophilic and thermophilic Group I archaea changed the century-old paradigm that aerobic ammonia oxidation is solely mediated by two small clades of Beta- and Gammaproteobacteria. Group I archaea are extremely diverse and ubiquitous in marine and terrestrial environments, accounting for 20-30% of the microbial plankton in the global oceans. Recent studies indicated that many of these organisms carry putative ammonia monooxygenase genes and are more abundant than ammonia-oxidizing bacteria in most natural environments suggesting a potentially significant role in the nitrogen cycle. The isolation of Nitrosopumilus maritimus strain SCM1 provided the first direct evidence that Group I archaea indeed gain energy from ammonia oxidation. To characterize the physiology of this archaeal nitrifier, we developed a respirometry setup particularly suited for activity measurements in dilute microbial cultures with extremely low oxygen uptake rates. Here, we describe the setup and review the kinetic experiments conducted with N. maritimus and other nitrifying microorganisms. These experiments demonstrated that N. maritimus is adapted to grow on ammonia concentrations found in oligotrophic open ocean environments, far below the survival threshold of ammonia-oxidizing bacteria. The described setup and experimental procedures should facilitate physiological studies on other nitrifying archaea and oligotrophic microorganisms in general. Copyright © 2011 Elsevier Inc. All rights reserved.

  17. Ammonia recovery from landfill leachate using hydrophobic membrane contactors.

    Science.gov (United States)

    Amaral, Míriam C S; Magalhães, Nátalie C; Moravia, Wagner G; Ferreira, Carolina D

    2016-11-01

    This article aims to evaluate membrane contactors capability to remove and recover ammonia from landfill leachate (LFL). A hydrophobic hollow fiber membrane module was used to achieve such purpose. A sulfuric acid diluted solution was used as extraction solution to speed up ammonia content removal. Several factors that have influence on ammonia removal and recovery capability such as ammonia solution pH, concentration of sulfuric acid solutions and flow rate of liquid phases have been examined. Microfiltration was the method used as pretreatment. The results have shown that membrane contactor operated with LFL (pH 10), 0.1 M acid solution and liquid flow rate up to 0.5 L min -1 achieved 99.9% of ammonia removal, which corresponds to 79.1% of ammonia recovery from the extraction solution, and it is capable to produce highly purified ammonium sulfate solutions (41.2%, wt wt -1 ) to be used as fertilizer. The concentration of total ammonia nitrogen (TAN) in the residual LFL complies with Brazilian law requirements of 20.0 mg L -1 of TAN, regarding the disposal of effluents.

  18. Investigations of blood ammonia analysis: Test matrices, storage, and stability.

    Science.gov (United States)

    Goldstein, Brittany N; Wesler, Jordan; Nowacki, Amy S; Reineks, Edmunds; Natowicz, Marvin R

    2017-06-01

    An assessment of blood ammonia concentration is common medical practice in the evaluation of an individual with an unexplained mental status change or coma. The determination of a blood ammonia level is most commonly done using a glutamate dehydrogenase (GLDH)-based assay, although there are many potential sources of artifact and the literature is inconsistent regarding key preanalytic issues. Using a GLDH-based assay, we first investigated matrix effects using three anticoagulants: heparin, EDTA and oxalate. Heparin-anticoagulated plasma was substantially less precise than EDTA- and oxalate-anticoagulated plasma. Oxalate-anticoagulated plasma showed a greater baseline of apparent ammonia than either heparin- or EDTA-derived plasma, presumably due to interferants. We then evaluated the stability of EDTA-anticoagulated plasma for assessment of ammonia when stored at 4°C,-14°C or -70°C. There was a linear increase of ammonia with storage at both 4°C and -14°C. Plasma kept at -70°C for up to three weeks showed no change in measured ammonia relative to the baseline determination. This work clarifies preanalytic conditions for which a precise determination of ammonia can be accomplished using a GLDH-based assay. Copyright © 2017 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  19. Ammonia removal in electrochemical oxidation: Mechanism and pseudo-kinetics

    International Nuclear Information System (INIS)

    Li Liang; Liu Yan

    2009-01-01

    This paper investigated the mechanism and pseudo-kinetics for removal of ammonia by electrochemical oxidation with RuO 2 /Ti anode using batch tests. The results show that the ammonia oxidation rates resulted from direct oxidation at electrode-liquid interfaces of the anode by stepwise dehydrogenation, and from indirect oxidation by hydroxyl radicals were so slow that their contribution to ammonia removal was negligible under the condition with Cl - . The oxidation rates of ammonia ranged from 1.0 to 12.3 mg N L -1 h -1 and efficiency reached nearly 100%, primarily due to the indirect oxidation of HOCl, and followed pseudo zero-order kinetics in electrochemical oxidation with Cl - . About 88% ammonia was removed from the solution. The removed one was subsequently found in the form of N 2 in the produced gas. The rate at which Cl - lost electrons at the anode was a major factor in the overall ammonia oxidation. Current density and Cl - concentration affected the constant of the pseudo zero-order kinetics, expressed by k = 0.0024[Cl - ] x j. The ammonia was reduced to less than 0.5 mg N L -1 after 2 h of electrochemical oxidation for the effluent from aerobic or anaerobic reactors which treated municipal wastewater. This result was in line with the strict discharge requirements

  20. Arterial ammonia levels in the management of fulminant liver failure

    Directory of Open Access Journals (Sweden)

    Curry S

    2011-06-01

    Full Text Available Previous studies have suggested that an arterial ammonia level greater than 150 mmol/L is highly sensitive for predicting subsequent development of cerebral edema in patients with fulminant liver failure. We performed a prospective cohort study to confirm this relationship. We enrolled 22 consecutive patients who presented to our transplant hepatology service with grade 3-4 encephalopathy associated with fulminant liver failure. All patients underwent placement of an intraparenchymal ICP monitor, and every 12 hourly arterial ammonia levels. The prevalence of intracranial hypertension (IHTN in our population was 95% (21/22 patients, with 82 discrete episodes recorded. The sensitivity of arterial ammonia levels to predict the onset of IHTN was 62% (95% CI: 40.8 to 79.3 at a cut point of 150 mmol/L. Arterial ammonia levels preceding the first intracranial hypertension event were less than 150 mmol/L in 8 of 21 patients (39%. Fifty nine of 82 episodes of IHTN (73% occurred when arterial ammonia levels were less than 150 mmol/L. We conclude that the arterial ammonia level is not useful in making decisions regarding management related to cerebral edema in patients with fulminant liver failure. In fact, since almost all our study patients with grade III or IV encephalopathy secondary to fulminant liver failure went on to develop intracranial hypertension, our study supports the contention that all such patients might benefit from ICP monitoring regardless of arterial ammonia levels.

  1. Safety assessment of ammonia as a transport fuel

    Energy Technology Data Exchange (ETDEWEB)

    Duijm, N.J.; Markert, F.; Lundtang paulsen, Jette

    2005-02-01

    This report describes the safety study performed as part of the EU supported project 'Ammonia Cracking for Clean Electric Power Technology' The study addresses the following activities: safety of operation of the ammonia-powered vehicle under normal and accident (collision) conditions, safety of transport of ammonia to the refuelling stations and safety of the activities at the refuelling station (unloading and refuelling). Comparisons are made between the safety of using ammonia and the safety of other existing or alternative fuels. The conclusion is that the hazards in relation to ammonia need to be controlled by a combination of technical and regulatory measures. The most important requirements are: - Advanced safety systems in the vehicle - Additional technical measures and regulations are required to avoid releases in maintenance workshops and unauthorised maintenance on the fuel system - Road transport of ammonia to refuelling stations in refrigerated form - Sufficient safety zones between refuelling stations and residential or otherwise public areas. When these measures are applied, the use of ammonia as a transport fuel wouldnt cause more risks than currently used fuels (using current practice). (au)

  2. Ammonia in the environment: From ancient times to the present

    International Nuclear Information System (INIS)

    Sutton, Mark A.; Erisman, Jan Willem; Dentener, Frank; Moeller, Detlev

    2008-01-01

    Recent research on atmospheric ammonia has made good progress in quantifying sources/sinks and environmental impacts. This paper reviews the achievements and places them in their historical context. It considers the role of ammonia in the development of agricultural science and air chemistry, showing how these arose out of foundations in 18th century chemistry and medieval alchemy, and then identifies the original environmental sources from which the ancients obtained ammonia. Ammonia is revealed as a compound of key human interest through the centuries, with a central role played by sal ammoniac in alchemy and the emergence of modern science. The review highlights how recent environmental research has emphasized volatilization sources of ammonia. Conversely, the historical records emphasize the role of high-temperature sources, including dung burning, coal burning, naturally burning coal seams and volcanoes. Present estimates of ammonia emissions from these sources are based on few measurements, which should be a future priority. - Past ammonia applications reveal new emphases in biospheric transformations

  3. Mathematical Model of Ammonia Handling in the Rat Renal Medulla

    Science.gov (United States)

    Noiret, Lorette; Baigent, Stephen; Jalan, Rajiv; Thomas, S. Randall

    2015-01-01

    The kidney is one of the main organs that produces ammonia and release it into the circulation. Under normal conditions, between 30 and 50% of the ammonia produced in the kidney is excreted in the urine, the rest being absorbed into the systemic circulation via the renal vein. In acidosis and in some pathological conditions, the proportion of urinary excretion can increase to 70% of the ammonia produced in the kidney. Mechanisms regulating the balance between urinary excretion and renal vein release are not fully understood. We developed a mathematical model that reflects current thinking about renal ammonia handling in order to investigate the role of each tubular segment and identify some of the components which might control this balance. The model treats the movements of water, sodium chloride, urea, NH3 and NH4+, and non-reabsorbable solute in an idealized renal medulla of the rat at steady state. A parameter study was performed to identify the transport parameters and microenvironmental conditions that most affect the rate of urinary ammonia excretion. Our results suggest that urinary ammonia excretion is mainly determined by those parameters that affect ammonia recycling in the loops of Henle. In particular, our results suggest a critical role for interstitial pH in the outer medulla and for luminal pH along the inner medullary collecting ducts. PMID:26280830

  4. SIRT5 regulation of ammonia-induced autophagy and mitophagy

    Science.gov (United States)

    Polletta, Lucia; Vernucci, Enza; Carnevale, Ilaria; Arcangeli, Tania; Rotili, Dante; Palmerio, Silvia; Steegborn, Clemens; Nowak, Theresa; Schutkowski, Mike; Pellegrini, Laura; Sansone, Luigi; Villanova, Lidia; Runci, Alessandra; Pucci, Bruna; Morgante, Emanuela; Fini, Massimo; Mai, Antonello; Russo, Matteo A; Tafani, Marco

    2015-01-01

    In liver the mitochondrial sirtuin, SIRT5, controls ammonia detoxification by regulating CPS1, the first enzyme of the urea cycle. However, while SIRT5 is ubiquitously expressed, urea cycle and CPS1 are only present in the liver and, to a minor extent, in the kidney. To address the possibility that SIRT5 is involved in ammonia production also in nonliver cells, clones of human breast cancer cell lines MDA-MB-231 and mouse myoblast C2C12, overexpressing or silenced for SIRT5 were produced. Our results show that ammonia production increased in SIRT5-silenced and decreased in SIRT5-overexpressing cells. We also obtained the same ammonia increase when using a new specific inhibitor of SIRT5 called MC3482. SIRT5 regulates ammonia production by controlling glutamine metabolism. In fact, in the mitochondria, glutamine is transformed in glutamate by the enzyme glutaminase, a reaction producing ammonia. We found that SIRT5 and glutaminase coimmunoprecipitated and that SIRT5 inhibition resulted in an increased succinylation of glutaminase. We next determined that autophagy and mitophagy were increased by ammonia by measuring autophagic proteolysis of long-lived proteins, increase of autophagy markers MAP1LC3B, GABARAP, and GABARAPL2, mitophagy markers BNIP3 and the PINK1-PARK2 system as well as mitochondrial morphology and dynamics. We observed that autophagy and mitophagy increased in SIRT5-silenced cells and in WT cells treated with MC3482 and decreased in SIRT5-overexpressing cells. Moreover, glutaminase inhibition or glutamine withdrawal completely prevented autophagy. In conclusion we propose that the role of SIRT5 in nonliver cells is to regulate ammonia production and ammonia-induced autophagy by regulating glutamine metabolism. PMID:25700560

  5. Dietary adipic acid reduces ammonia emission from swine excreta.

    Science.gov (United States)

    van Kempen, T A

    2001-09-01

    Adipic acid is only partially catabolized when it is fed to animals, and a portion of it is excreted in urine. The excreted portion may lower urinary pH and, as a result, ammonia emission. The present study tested this hypothesis. In Exp. 1, nursery pigs (n = 14) were fed (for a period of 7 d) either a standard nursery diet or the same diet supplemented with 1% adipic acid to assess effects on urinary pH (collected on d 5 or 6) and in vitro ammonia emission from the collected urine samples that were mixed with control feces. In Exp. 2, grower pigs housed 10 each in one of two chambers were fed a control diet or the same diet supplemented with 1% adipic acid. Ventilated air was quantified and analyzed for ammonia using Fourier transform infrared spectroscopy to determine the effects of feeding 1% adipic acid on ammonia emission. The results from Exp. 1 showed that adipic acid strongly reduced urinary pH (from 7.7 to 5.5, P < 0.05). In vitro ammonia emission from these urine samples was significantly reduced at all the time points evaluated (1, 3, 18, and 46 h with reductions of 94, 93, 70, and 39%, respectively, P < 0.05). Experiment 2 showed that adipic acid supplementation reduced ammonia emission by 25% (P < 0.05), which corresponded to the predicted reduction in ammonia emission based on the reduction in manure pH observed. In conclusion, feeding adipic acid lowers urinary pH and reduces ammonia emission. The reduction in ammonia emission, though, does not correspond to the reduction in urinary pH but corresponds to the reduction in fecal pH as a result of mixing the urine and feces, in which feces act as a strong buffer.

  6. Dry anaerobic ammonia-methane production from chicken manure.

    Science.gov (United States)

    Abouelenien, Fatma; Kitamura, Yoshiaki; Nishio, Naomichi; Nakashimada, Yutaka

    2009-03-01

    The effect of temperature on production of ammonia during dry anaerobic fermentation of chicken manure (CM), inoculated with thermophilic methanogenic sludge, was investigated in a batch condition for 8 days. Incubation temperature did not have a significant effect on the production of ammonia. Almost complete inhibition of production of methane occurred at 55 and 65 degrees C while quite low yields of 8.45 and 6.34 ml g(-1) VS (volatile solids) were observed at 35 and 45 degrees C due to a higher accumulation of ammonia. In order to improve the production of methane during dry anaerobic digestion of CM, stripping of ammonia was performed firstly on the CM previously fermented at 65 degrees C for 8 days: the stripping for 1 day at 85 degrees C and pH 10 removed 85.5% of ammonia. The first-batch fermentation of methane for 75 days was conducted next, using the ammonia-stripped CM inoculated with methanogenic sludge at different ratios, (CM: thermophilic sludge) of 1:2, 1:1, and 2:1 on volume per volume basis at both 35 and 55 degrees C. Production of methane improved and was higher than that of the control (without stripping of ammonia) but the yield of 20.4 ml g(-1) VS was still low, so second stripping of ammonia was conducted, which resulted in 74.7% removal of ammonia. A great improvement in the production of methane of 103.5 ml g(-1) VS was achieved during the second batch for 55 days.

  7. The effect of gaseous ammonia on cobalt perrhenate

    International Nuclear Information System (INIS)

    Maslov, L.P.; Men'shikov, O.D.; Borisov, V.V.; Sorokin, S.I.; Krutovertsev, S.A.; Kharkevich, S.I.; Ivanova, O.M.

    1994-01-01

    The influence of humid air ammonia mixture on crystal pentahydrate of cobalt(2) perrhenate has been studied by the methods of PES, IR spectroscopy thermal analysis and electrophysical measurements. It is shown that with an increase in ammonia content in gaseous phase cobalt perrhenate successively transforms into diaquodiammine-, tetrammine- and μ-dioxo-bis-(tetrammine) derivatives of cobalt. Reversibility of dioxocomplex formation and a correlation between the change in electrophysical properties of crystal sample and change in ammonia content in gaseous phase are pointed out. 16 refs.; 4 figs.; 1 tab

  8. Erbium Doped Fiber Sensor for Ammonia Detection into Water

    International Nuclear Information System (INIS)

    Pérez-Sánchez, G G; Pinzón-Escobar, E F; Sandoval-Romero, G E; Chávez, J A Álvarez

    2015-01-01

    The water pollution is a health problem around the world. One of the most common pollutants in water is the Ammonia. Some sensors have been developed for Ammoniac detection even though most of them are not in real time and could be expensive. In this work an Erbium-doped fibre-sensor for Ammonia detection with a mechanical long period fibre grating and a taper in cascade for improving the sensitivity; the detection bandwidth is in the 1550 nm region from 1460 nm to 1640 nm. Output optical spectrum for Ammonia concentrations from 1ml to 5 ml is shown

  9. The Sugar Model: Autocatalytic Activity of the Triose Ammonia Reaction

    Science.gov (United States)

    Weber, Arthur L.

    2007-04-01

    Reaction of triose sugars with ammonia under anaerobic conditions yielded autocatalytic products. The autocatalytic behavior of the products was examined by measuring the effect of the crude triose ammonia reaction product on the kinetics of a second identical triose ammonia reaction. The reaction product showed autocatalytic activity by increasing both the rate of disappearance of triose and the rate of formation of pyruvaldehyde, the product of triose dehydration. This synthetic process is considered a reasonable model of origin-of-life chemistry because it uses plausible prebiotic substrates, and resembles modern biosynthesis by employing the energized carbon groups of sugars to drive the synthesis of autocatalytic molecules.

  10. Catalytic synthesis of ammonia using vibrationally excited nitrogen

    DEFF Research Database (Denmark)

    Henriksen, Niels Engholm; Billing, Gert D.; Hansen, Flemming Yssing

    1992-01-01

    In a previous study we have considered the catalytic synthesis of ammonia in the presence of vibrationally excited nitrogen. The distribution over vibrational states was assumed to be maintained during the reaction, and it was shown that the yield of ammonia increased considerably compared...... to that from conventional synthesis. In the present study the nitrogen molecules are only excited at the inlet of a plug flow reactor, and the importance of vibrational relaxation is investigated. We show that vibrational excitation can give an enhanced yield of ammonia also in the situation where vibrational...

  11. Crystallization and preliminary X-ray analysis of l-methionine γ-lyase 1 from Entamoeba histolytica

    Energy Technology Data Exchange (ETDEWEB)

    Sato, Dan [Institute for Advanced Biosciences, Keio University, 246-2 Mizukami, Kakuganji, Tsuruoka, Yamagata 997-0052 (Japan); Center for Integrated Medical Research, School of Medicine, Keio University, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582 (Japan); Department of Parasitology, Gunma University Graduate School of Medicine, 3-39-22 Showa-machi, Maebashi, Gunma 371-8511 (Japan); Karaki, Tsuyoshi; Shimizu, Akira; Kamei, Kaeko; Harada, Shigeharu, E-mail: harada@kit.ac.jp [Graduate School of Science and Technology, Department of Applied Biology, Kyoto Institute of Technology, Sakyo-ku, Kyoto 606-8585 (Japan); Nozaki, Tomoyoshi [Department of Parasitology, Gunma University Graduate School of Medicine, 3-39-22 Showa-machi, Maebashi, Gunma 371-8511 (Japan); Department of Parasitology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640 (Japan); Institute for Advanced Biosciences, Keio University, 246-2 Mizukami, Kakuganji, Tsuruoka, Yamagata 997-0052 (Japan)

    2008-08-01

    l-Methionine γ-lyase 1, a key enzyme in sulfur-containing amino-acid degradation, from the protozoan parasite E. histolytica was crystallized in a form suitable for X-ray structure analysis. l-Methionine γ-lyase (MGL) is a pyridoxal phosphate-dependent enzyme that is involved in the degradation of sulfur-containing amino acids. MGL is an attractive drug target against amoebiasis because the mammalian host of its causative agent Entamoeba histolytica lacks MGL. For the development of anti-amoebic agents based on the structure of MGL, one of two MGL isoenzymes (EhMGL1) was crystallized in the monoclinic space group P2{sub 1}, with unit-cell parameters a = 99.12, b = 85.38, c = 115.37 Å, β = 101.82°. The crystals diffract to beyond 2.0 Å resolution. The presence of a tetramer in the asymmetric unit (4 × 42.4 kDa) gives a Matthews coefficient of 2.8 Å{sup 3} Da{sup −1} and a solvent content of 56%. The structure was solved by the molecular-replacement method and structure refinement is now in progress.

  12. Adaptation to Blue Light in MarineSynechococcusRequires MpeU, an Enzyme with Similarity to Phycoerythrobilin Lyase Isomerases.

    Science.gov (United States)

    Mahmoud, Rania M; Sanfilippo, Joseph E; Nguyen, Adam A; Strnat, Johann A; Partensky, Frédéric; Garczarek, Laurence; Abo El Kassem, Nabil; Kehoe, David M; Schluchter, Wendy M

    2017-01-01

    Marine Synechococcus has successfully adapted to environments with different light colors, which likely contributes to this genus being the second most abundant group of microorganisms worldwide. Populations of Synechococcus that grow in deep, blue ocean waters contain large amounts of the blue-light absorbing chromophore phycourobilin (PUB) in their light harvesting complexes (phycobilisomes). Here, we show that all Synechococcus strains adapted to blue light possess a gene called mpeU . MpeU is structurally similar to phycobilin lyases, enzymes that ligate chromophores to phycobiliproteins. Interruption of mpeU caused a reduction in PUB content, impaired phycobilisome assembly and reduced growth rate more strongly in blue than green light. When mpeU was reintroduced in the mpeU mutant background, the mpeU- less phenotype was complemented in terms of PUB content and phycobilisome content. Fluorescence spectra of mpeU mutant cells and purified phycobilisomes revealed red-shifted phycoerythrin emission peaks, likely indicating a defect in chromophore ligation to phycoerythrin-I (PE-I) or phycoerythrin-II (PE-II). Our results suggest that MpeU is a lyase-isomerase that attaches a phycoerythrobilin to a PEI or PEII subunit and isomerizes it to PUB. MpeU is therefore an important determinant in adaptation of Synechococcus spp. to capture photons in blue light environments throughout the world's oceans.

  13. Ability of phages to infect Acinetobacter calcoaceticus-Acinetobacter baumannii complex species through acquisition of different pectate lyase depolymerase domains.

    Science.gov (United States)

    Oliveira, Hugo; Costa, Ana R; Konstantinides, Nico; Ferreira, Alice; Akturk, Ergun; Sillankorva, Sanna; Nemec, Alexandr; Shneider, Mikhail; Dötsch, Andreas; Azeredo, Joana

    2017-12-01

    Bacteriophages are ubiquitous in nature and represent a vast repository of genetic diversity, which is driven by the endless coevolution cycle with a diversified group of bacterial hosts. Studying phage-host interactions is important to gain novel insights into their dynamic adaptation. In this study, we isolated 12 phages infecting species of the Acinetobacter baumannii-Acinetobacter calcoaceticus complex which exhibited a narrow host range and similar morphological features (podoviruses with short tails of 9-12 nm and isometric heads of 50-60 nm). Notably, the alignment of the newly sequenced phage genomes (40-41 kb of DNA length) and all Acinetobacter podoviruses deposited in Genbank has shown high synteny, regardless of the date and source of isolation that spans from America to Europe and Asia. Interestingly, the C-terminal pectate lyase domain of these phage tail fibres is often the only difference found among these viral genomes, demonstrating a very specific genomic variation during the course of their evolution. We proved that the pectate lyase domain is responsible for phage depolymerase activity and binding to specific Acinetobacter bacterial capsules. We discuss how this mechanism of phage-host co-evolution impacts the tail specificity apparatus of Acinetobacter podoviruses. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

  14. Submersible microbial desalination cell for simultaneous ammonia recovery and electricity production from anaerobic reactors containing high levels of ammonia.

    Science.gov (United States)

    Zhang, Yifeng; Angelidaki, Irini

    2015-02-01

    High ammonia concentration in anaerobic reactors can seriously inhibit the anaerobic digestion process. In this study, a submersible microbial desalination cell (SMDC) was developed as an innovative method to lower the ammonia level in a continuous stirred tank reactor (CSTR) by in situ ammonia recovery and electricity production. In batch experiment, the ammonia concentration in the CSTR decreased from 6 to 0.7 g-N/L during 30 days, resulting in an average recovery rate of 80 g-N/m(2)/d. Meanwhile, a maximum power density of 0.71±0.5 W/m(2) was generated at 2.85 A/m(2). Both current driven NH4(+) migration and free NH3 diffusion were identified as the mechanisms responsible for the ammonia transportation. With an increase in initial ammonia concentration and a decrease in external resistance, the SMDC performance was enhanced. In addition, the coexistence of other cations in CSTR or cathode had no negative effect on the ammonia transportation. Copyright © 2014 Elsevier Ltd. All rights reserved.

  15. Mechanistic Studies of the Spore Photoproduct Lyase (SPL) via a Single Cysteine Mutation

    Science.gov (United States)

    Yang, Linlin; Lin, Gengjie; Nelson, Renae S.; Jian, Yajun; Telser, Joshua; Li, Lei

    2012-01-01

    5-thyminyl-5,6-dihydrothymine (also called spore photoproduct or SP) is the exclusive DNA photo-damage product in bacterial endospores. It is repaired by a radical SAM (S-adenosylmethionine) enzyme, the spore photoproduct lyase (SPL), at the bacterial early germination phase. Our previous studies proved that SPL utilizes the 5′-dA• generated by SAM cleavage reaction to abstract the H6proR atom to initiate the SP repair process. The resulting thymine allylic radical was suggested to take an H atom from an unknown protein source, most likely the cysteine 141. Here we show that C141 can be readily alkylated in the native SPL by iodoacetamide treatment, suggesting that it is accessible to the TpT radical. SP repair by the SPL C141A mutant yields TpTSO2− and TpT simultaneously from the very beginning of the reaction; no lag phase is observed for the TpTSO2− formation. Should any other protein residue serve as the H donor, its presence would result in TpT as the major product at least for the first enzyme turnover. These observations provide strong evidence to support C141 as the direct H atom donor. Moreover, due to the lack of this intrinsic H donor, the C141A mutant produces TpT via an unprecedented thymine cation radical reduction (proton coupled electron transfer) process, contrasting to the H atom transfer mechanism in the WT SPL reaction. The C141A mutant repairs SP at a rate which is ~3-fold slower than the WT enzyme. Formation of TpTSO2− and TpT exhibit a Vmax deuterium kinetic isotope effect (KIE) of 1.7 ± 0.2 respectively, which is smaller than the DVmax KIE of 2.8 ± 0.3 determined in the WT SPL reaction. These findings suggest that removing the intrinsic H atom donor disturbs the rate-limiting process in the enzyme catalysis. As expected, the pre-reduced C141A mutant only supports ~ 0.4 turnover, which is in sharp contrast to the > 5 turnovers exhibited by the WT SPL reaction, suggesting that the enzyme catalytic cycle (SAM regeneration) is

  16. Ammonia removal from an aqueous solution and method for the production of a fertilizer material

    NARCIS (Netherlands)

    Kelder, E.M.; Ursem, W.N.J.; Roos, R.A.; Marijnissen, J.C.M.

    2011-01-01

    The invention provides method for the removal of ammonia from air and converting the ammonia in a fertilizer material, comprising (a) contacting at least part of the air with an aqueous liquid, wherein the aqueous liquid is a solvent for ammonia, to provide an ammonia containing aqueous

  17. 46 CFR 111.105-32 - Bulk liquefied flammable gas and ammonia carriers.

    Science.gov (United States)

    2010-10-01

    ... 46 Shipping 4 2010-10-01 2010-10-01 false Bulk liquefied flammable gas and ammonia carriers. 111... gas and ammonia carriers. (a) Each vessel that carries bulk liquefied flammable gases or ammonia as a.... (2) The term “gas-dangerous” does not include the weather deck of an ammonia carrier. (c) Each...

  18. New efficient catalyst for ammonia synthesis: barium-promoted cobalt on carbon

    DEFF Research Database (Denmark)

    Hagen, Stefan; Barfod, Rasmus; Fehrmann, Rasmus

    2002-01-01

    Barium-promoted cobalt catalysts supported on carbon exhibit higher ammonia activities at synthesis temperatures than the commercial, multipromoted iron catalyst and also a lower ammonia......Barium-promoted cobalt catalysts supported on carbon exhibit higher ammonia activities at synthesis temperatures than the commercial, multipromoted iron catalyst and also a lower ammonia...

  19. Ammonia stress under high environmental ammonia induces Hsp70 and Hsp90 in the mud eel, Monopterus cuchia.

    Science.gov (United States)

    Hangzo, Hnunlalliani; Banerjee, Bodhisattwa; Saha, Shrabani; Saha, Nirmalendu

    2017-02-01

    The obligatory air-breathing mud eel (Monopterus cuchia) is frequently being challenged with high environmental ammonia (HEA) exposure in its natural habitats. The present study investigated the possible induction of heat shock protein 70 and 90 (hsp70, hsc70, hsp90α and hsp90β) genes and more expression of Hsp70 and Hsp90 proteins under ammonia stress in different tissues of the mud eel after exposure to HEA (50 mM NH 4 Cl) for 14 days. HEA resulted in significant accumulation of toxic ammonia in different body tissues and plasma, which was accompanied with the stimulation of oxidative stress in the mud eel as evidenced by more accumulation of malondialdehyde (MDA) and hydrogen peroxide (H 2 O 2 ) during exposure to HEA. Further, hyper-ammonia stress led to significant increase in the levels of mRNA transcripts for inducible hsp70 and hsp90α genes and also their translated proteins in different tissues probably as a consequence of induction of hsp70 and hsp90α genes in the mud eel. However, hyper-ammonia stress was neither associated with any significant alterations in the levels of mRNA transcripts for constitutive hsc70 and hsp90β genes nor their translated proteins in any of the tissues studied. More abundance of Hsp70 and Hsp90α proteins might be one of the strategies adopted by the mud eel to defend itself from the ammonia-induced cellular damages under ammonia stress. Further, this is the first report of ammonia-induced induction of hsp70 and hsp90α genes under hyper-ammonia stress in any freshwater air-breathing teleost.

  20. Risk analysis and emergency management of ammonia installations

    NARCIS (Netherlands)

    Ham, J.M.; Gansevoort, J.

    1992-01-01

    The use of Quantitative Risk Assessment has been increasing for evaluating the risk of handling hazardous materials and land-use planning. This article reports on several studies carried out on the risk of handling, storage and transport of ammonia.

  1. Development of High-Performance Ammonia Borane Based Rocket Propellants

    Data.gov (United States)

    National Aeronautics and Space Administration — Ammonia borane based fuels for use in hybrid rocket systems have the potential to be high performing while at the same time mitigating many of the issues associated...

  2. Different cultivation methods to acclimatise ammonia-tolerant methanogenic consortia

    DEFF Research Database (Denmark)

    Tian, Hailin; Fotidis, Ioannis; Mancini, Enrico

    2017-01-01

    Bioaugmentation with ammonia tolerant-methanogenic consortia was proposed as a solution to overcome ammonia inhibition during anaerobic digestion process recently. However, appropriate technology to generate ammonia tolerant methanogenic consortia is still lacking. In this study, three basic...... reactors (i.e. batch, fed-batch and continuous stirred-tank reactors (CSTR)) operated at mesophilic (37°C) and thermophilic (55°C) conditions were assessed, based on methane production efficiency, incubation time, TAN/FAN (total ammonium nitrogen/free ammonia nitrogen) levels and maximum methanogenic...... activity. Overall, fed-batch cultivation was clearly the most efficient method compared to batch and CSTR. Specifically, by saving incubation time up to 150%, fed-batch reactors were acclimatised to nearly 2-fold higher FAN levels with a 37%-153% methanogenic activity improvement, compared to batch method...

  3. Ammonia effect on hydrogenotrophic methanogens and syntrophic acetate oxidizing bacteria

    DEFF Research Database (Denmark)

    Wang, Han; Fotidis, Ioannis; Angelidaki, Irini

    methanogens. Two pure strains of hydrogenotrophic methanogens (i.e: Methanoculleus bourgensis and Methanoculleus thermophiles) and two pure strains of SAO bacteria (i.e: Tepidanaerobacter acetatoxydans and Thermacetogenium phaeum) were inoculated under four different ammonia (0.26, 3, 5 and 7g NH4+-N....../L) and free ammonia levels (Mesophilic: 3.31, 38.2, 63.68 and 89.15 g NH3-N/L. Thermophilic: 8.48, 97.82, 163.03 and 228.24 g NH3-N/L). The results indicated that both T. acetatoxydans and T. phaeum were more sensitive to high ammonia levels compared to the hydrogenotrophic methanogens tested. Additionally......, the total incubation periods of hydrogenotrophic methanogens were significantly shorter compared to the SAO bacteria incubation periods. Thus, it seems that hydrogenotrophic methanogens could be equally, if not more, tolerant to high ammonia levels compared to SAO bacteria....

  4. Computation of Equilibrium Partial Vapor Pressures of Aqueous Ammonia Solutions

    National Research Council Canada - National Science Library

    Field, Paul

    2000-01-01

    This report evaluates all available vapor-liquid equilibria (VLE) data from low-to-moderate pressures over the normal liquid water temperature range for the highly non-ideal aqueous ammonia solutions spanning nearly a century...

  5. The production of ammonia by multiheme cytochromes C.

    Science.gov (United States)

    Simon, Jörg; Kroneck, Peter M H

    2014-01-01

    The global biogeochemical nitrogen cycle is essential for life on Earth. Many of the underlying biotic reactions are catalyzed by a multitude of prokaryotic and eukaryotic life forms whereas others are exclusively carried out by microorganisms. The last century has seen the rise of a dramatic imbalance in the global nitrogen cycle due to human behavior that was mainly caused by the invention of the Haber-Bosch process. Its main product, ammonia, is a chemically reactive and biotically favorable form of bound nitrogen. The anthropogenic supply of reduced nitrogen to the biosphere in the form of ammonia, for example during environmental fertilization, livestock farming, and industrial processes, is mandatory in feeding an increasing world population. In this chapter, environmental ammonia pollution is linked to the activity of microbial metalloenzymes involved in respiratory energy metabolism and bioenergetics. Ammonia-producing multiheme cytochromes c are discussed as paradigm enzymes.

  6. Process for producing 13N-ammonia and device therefor

    International Nuclear Information System (INIS)

    Tanaka, Akira; Suzuki, Hirofumi.

    1997-01-01

    Ethanol or hydrogen is added to purified water or distilled water for injection, they are filled into a target vessel, and proton beams are irradiated to produce 13N-ammonia in the target vessel. A target liquid containing the resultant is introduced to a heat-reaction vessel. A slight amount of weak alkali solution is added to the target liquid in the heat-reaction vessel. The reaction vessel is heated to evaporate water and 13N-ammonia, and they are transferred to a vial. In this case, nitrogen gas as a gas to be entrained is supplied. 13N-ammonia is subjected to bubbling into the distilled water for injection or physiological saline water and recovered into the vial. 13N-ammonia is thus separated and purified as an injection which is a medical labelled compound. (I.N.)

  7. CO2 emissions and mitigation potential in China's ammonia industry

    International Nuclear Information System (INIS)

    Zhou Wenji; Zhu Bing; Li Qiang; Ma Tieju; Hu Shanying; Griffy-Brown, Charla

    2010-01-01

    Significant pressure from increasing CO 2 emissions and energy consumption in China's industrialization process has highlighted a need to understand and mitigate the sources of these emissions. Ammonia production, as one of the most important fundamental industries in China, represents those heavy industries that contribute largely to this sharp increasing trend. In the country with the largest population in the world, ammonia output has undergone fast growth spurred by increasing demand for fertilizer of food production since 1950s. However, various types of technologies implemented in the industry make ammonia plants in China operate with huge differences in both energy consumption and CO 2 emissions. With consideration of these unique features, this paper attempts to estimate the amount of CO 2 emission from China's ammonia production, and analyze the potential for carbon mitigation in the industry. Based on the estimation, related policy implications and measures required to realize the potential for mitigation are also discussed.

  8. Biokinetic characterization of the acceleration phase in autotrophic ammonia oxidation.

    Science.gov (United States)

    Chandran, Kartik; Smets, Barth F

    2008-08-01

    Batch autotrophic ammonia oxidation tracked through oxygen uptake measurements displays a preliminary acceleration phase. Failure to recognize the acceleration phase and fitting batch ammonia oxidation profiles with standard Monod-type mathematical models can result in meaningless kinetic parameter estimates. The objectives of this study were to examine the factors controlling the acceleration phase and to derive and test empirical and metabolic models for its description. Because of possible sustained reducing power limitation during batch ammonia oxidation, the extent of the acceleration phase (1) increased with increasing initial ammonia concentration, (2) did not systematically vary with initial biomass concentrations, and (3) increased in response to starvation. Concurrent hydroxylamine oxidation significantly reduced the acceleration phase potentially by relieving reducing power limitation. A nonlinear empirical model described the acceleration phase more accurately than a linear empirical model. The metabolic model also captured experimental trends exceedingly well, but required determination of additional parameters and variables.

  9. Hydrogen Production from Ammonia Using a Plasma Membrane Reactor

    Directory of Open Access Journals (Sweden)

    Shinji Kambara

    2016-06-01

    Full Text Available In this study, an efficient method for using pulsed plasma to produce hydrogen from ammonia was developed. An original pulsed plasma reactor with a hydrogen separation membrane was developed for efficient hydrogen production, and its hydrogen production performance was investigated. Hydrogen production in the plasma was affected by the applied voltage and flow rate of ammonia gas. The maximum hydrogen production flow rate of a typical plasma reactor was 8.7 L/h, whereas that of the plasma membrane reactor was 21.0 L/h. We found that ammonia recombination reactions in the plasma controlled hydrogen production in the plasma reactor. In the plasma membrane reactor, a significant increase in hydrogen production was obtained because ammonia recombination reactions were inhibited by the permeation of hydrogen radicals generated in the plasma through a palladium alloy membrane. The energy efficiency was 4.42 mol-H2/kWh depending on the discharge power.

  10. Predicting catalysis: Understanding ammonia synthesis from first-principles calculations

    DEFF Research Database (Denmark)

    Hellmann, A.; Baerends, E.J.; Biczysko, M.

    2006-01-01

    . Furthermore, our studies provide new insight into several related fields, for instance, gas-phase and electrochemical ammonia synthesis. The success of predicting the outcome of a catalytic reaction from first-principles calculations supports our point of view that, in the future, theory will be a fully......Here, we give a full account of a large collaborative effort toward an atomic-scale understanding of modern industrial ammonia production over ruthenium catalysts. We show that overall rates of ammonia production can be determined by applying various levels of theory (including transition state...... for any given point along an industrial reactor, and the kinetic results can be integrated over the catalyst bed to determine the industrial reactor yield. We find that, given the present uncertainties, the rate of ammonia production is well-determined directly from our atomic-scale calculations...

  11. Review of Options for Ammonia/Ammonium Management

    Energy Technology Data Exchange (ETDEWEB)

    Nash, C. A. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)

    2016-05-06

    This report is a review of literature supporting practical ammonia/ammonium destruction processes. Melter research supporting Hanford Low Activity Waste (LAW) glass production has shown that significant amounts of ammonia will be in the melter offgas condensate. Further work with secondary waste forms indicates the potential need to remove the ammonia, perhaps by an oxidative process. This review finds likely practical chemical methods to oxidize ammonia in aqueous solution at moderate temperatures and atmospheric pressure, using easily obtained reagents. Leading candidates include nitrite oxidation to produce nitrogen gas, various peroxide oxidative processes, and air stripping. This work reviews many other processes and provides reasoning to not consider those processes further for this application.

  12. A process to remove ammonia from PUREX plant effluents

    International Nuclear Information System (INIS)

    Moore, J.D.

    1990-01-01

    Zirconium-clad nuclear fuel from the Hanford N-Reactor is reprocessed in the PUREX (Plutonium Uranium Extraction) Plant operated by the Westinghouse Hanford Comapny. Before dissolution, cladding is chemically removed from the fuel elements with a solution of ammonium fluoride-ammonium nitrate (AFAN). a solution batch with an ammonia equivalent of about 1,100 kg is added to each fuel batch of 10 metric tons. This paper reports on this decladding process, know as the 'Zirflex' process which produces waste streams containing ammonia and ammonium slats. Waste stream treatment, includes ammonia scrubbing, scrub solution evaporation, residual solids dissolution, and chemical neutralization. These processes produce secondary liquid and gaseous waste streams containing varying concentrations of ammonia and low-level concentrations of radionuclides. Until legislative restrictions were imposed in 1987, these secondary streams were released to the soil in a liquid disposal 'crib' and to the atmosphere

  13. Ammonia removal via microbial fuel cell (MFC) dynamic reactor

    Science.gov (United States)

    Alabiad, I.; Ali, U. F. M.; Zakarya, I. A.; Ibrahim, N.; Radzi, R. W.; Zulkurnai, N. Z.; Azmi, N. H.

    2017-06-01

    Landfill leachate is generally known as high-strength wastewater that is difficult to handle and contains dissolved extracts and suspended matter. Microbial fuel cells (MFCs) were designed to treat landfill leachate while continuously producing power (voltage output). Three different anodes were tested in MFC reactors: carbon black, activated carbon, and zinc electrodes. Movements in the MFC reactor during treatment were also a key factor for testing. Results showed a difference in ammonia levels in the three anodes used. The study compared the efficiency of static and dynamic modes of MFC in removing ammonia. Continual leachate movement in the reactor could increase the rate of removal of the ammonia components. The setup provided a viable condition for maximum removal because the reactor movement caused the sludge to disintegrate, which allowed ammonia to separate easily from the parent leachate. Ammonia removal also resulted from the transfer of ammonium through the membrane or from ammonia loss. Constant exchange of ionic content benefited the MFC performance by increasing power production and decreasing internal electrode material resistance. This paper presents the results of the analyses of leachate treatment from the solid waste landfill located in Padang Siding Landfill, Perlis. The performance of ammonia removal was enhanced using different types of electrodes. In both modes, activated carbon performed better than black carbon and zinc. The respective percentages of ammonia removal for activated carbon of dynamic over static were 96.6%, 66.6%, and 92.8% for activated carbon, zinc, and black carbon. The results provide further information on the possibility of using MFCs in landfill leachate treatment systems.

  14. Exhaust purification with on-board ammonia production

    Science.gov (United States)

    Robel, Wade J [Peoria, IL; Driscoll, James Joshua [Dunlap, IL; Coleman, Gerald N [Peterborough, GB

    2008-05-13

    A system of ammonia production for a selective catalytic reduction system is provided. The system includes producing an exhaust gas stream within a cylinder group, wherein the first exhaust gas stream includes NOx. The exhaust gas stream may be supplied to an exhaust passage and cooled to a predetermined temperature range, and at least a portion of the NOx within the exhaust gas stream may be converted into ammonia.

  15. R-32 As An Alternative To Ammonia In Industrial Refrigeration

    OpenAIRE

    Pearson, Andy

    2016-01-01

    Ammonia is recognised to be the most commonly used refrigerant in industrial systems however it is limited in some applications by its toxicity. Â R-32 has similar flammability characteristics and a similar pressure-temperature relationship, and through its use as a blend component it has become widely used and readily available. Â This paper compares and contrasts ammonia and R-32 with specific reference to industrial applications. Â The analysis includes comparison of refrigerating effect, ...

  16. Calcium in the mechanism of ammonia-induced astrocyte swelling.

    Science.gov (United States)

    Jayakumar, Arumugam R; Rama Rao, Kakulavarapu V; Tong, Xiaoying Y; Norenberg, Michael D

    2009-05-01

    Brain edema, due largely to astrocyte swelling, is an important clinical problem in patients with acute liver failure. While mechanisms underlying astrocyte swelling in this condition are not fully understood, ammonia and associated oxidative/nitrosative stress appear to be involved. Mechanisms responsible for the increase in reactive oxygen/nitrogen species (RONS) and their role in ammonia-induced astrocyte swelling, however, are poorly understood. Recent studies have demonstrated a transient increase in intracellular Ca2+ in cultured astrocytes exposed to ammonia. As Ca2+ is a known inducer of RONS, we investigated potential mechanisms by which Ca2+ may be responsible for the production of RONS and cell swelling in cultured astrocytes after treatment with ammonia. Exposure of cultured astrocytes to ammonia (5 mM) increased the formation of free radicals, including nitric oxide, and such increase was significantly diminished by treatment with the Ca2+ chelator 1,2-bis-(o-aminophenoxy)-ethane-N,N,-N',N'-tetraacetic acid tetraacetoxy-methyl ester (BAPTA). We then examined the activity of Ca2+-dependent enzymes that are known to generate RONS and found that ammonia significantly increased the activities of NADPH oxidase (NOX), constitutive nitric oxide synthase (cNOS), and phospholipase A2 (PLA2) and such increases in activity were significantly diminished by BAPTA. Pre-treatment of cultures with 7-nitroindazole, apocyanin, and quinacrine, respective inhibitors of cNOS, NOX, and PLA2, all significantly diminished RONS production. Additionally, treatment of cultures with BAPTA or with inhibitors of cNOS, NOX, and PLA2 reduced ammonia-induced astrocyte swelling. These studies suggest that the ammonia-induced rise in intracellular Ca2+ activates free radical producing enzymes that ultimately contribute to the mechanism of astrocyte swelling.

  17. Ammonia concentration modeling based on retained gas sampler data

    International Nuclear Information System (INIS)

    Terrones, G.; Palmer, B.J.; Cuta, J.M.

    1997-09-01

    The vertical ammonia concentration distributions determined by the retained gas sampler (RGS) apparatus were modeled for double-shell tanks (DSTs) AW-101, AN-103, AN-104, and AN-105 and single-shell tanks (SSTs) A-101, S-106, and U-103. One the vertical transport of ammonia in the tanks were used for the modeling. Transport in the non-convective settled solids and floating solids layers is assumed to occur primarily via some type of diffusion process, while transport in the convective liquid layers is incorporated into the model via mass transfer coefficients based on empirical correlations. Mass transfer between the top of the waste and the tank headspace and the effects of ventilation of the headspace are also included in the models. The resulting models contain a large number of parameters, but many of them can be determined from known properties of the waste configuration or can be estimated within reasonable bounds from data on the waste samples themselves. The models are used to extract effective diffusion coefficients for transport in the nonconvective layers based on the measured values of ammonia from the RGS apparatus. The modeling indicates that the higher concentrations of ammonia seen in bubbles trapped inside the waste relative to the ammonia concentrations in the tank headspace can be explained by a combination of slow transport of ammonia via diffusion in the nonconvective layers and ventilation of the tank headspace by either passive or active means. Slow transport by diffusion causes a higher concentration of ammonia to build up deep within the waste until the concentration gradients between the interior and top of the waste are sufficient to allow ammonia to escape at the same rate at which it is being generated in the waste

  18. Predicting Catalysis: Understanding Ammonia Synthesis from First-Principles Calculations

    OpenAIRE

    Hellman, A.; Baerends, E. J.; Biczysko, M.; Bligaard, T.; Christensen, C. H.; Clary, D. C.; Dahl, S.; Harrevelt, R. van; Honkala, K.; Jonsson, H.; Kroes, G. J.; Luppi, M.; Manthe, U.; Nørskov, J. K.; Olsen, R. A.

    2006-01-01

    Here, we give a full account of a large collaborative effort toward an atomic-scale understanding of modern industrial ammonia production over ruthenium catalysts. We show that overall rates of ammonia production can be determined by applying various levels of theory (including transition state theory with or without tunneling corrections, and quantum dynamics) to a range of relevant elementary reaction steps, such as N2 dissociation, H2 dissociation, and hydrogenation of the intermediate rea...

  19. Time Delay for Aerial Ammonia Concentration Measurements in Livestock Buildings

    OpenAIRE

    Rom; Zhang

    2010-01-01

    Correct measurements of ammonia concentration in air still present considerable challenges. The high water solubility and polarity can cause it to adsorb on surfaces in the entire sampling system, including sampling lines, filters, valves, pumps and instruments, causing substantial measuring errors and time delays. To estimate time delay characteristics of a Photo Acoustic Multi Gas Monitor 1312 and a Multi Point Sampler continuous measurement of aerial ammonia concentrations at different lev...

  20. Isotopic Discrimination of Some Solutes in Liquid Ammonia

    Science.gov (United States)

    Taube, H.; Viste, A.

    1966-01-01

    The nitrogen isotopic discrimination of some salts and metals, studies in liquid ammonia solution at -50�C, decreases in magnitude in the order Pb{sup ++}, Ca{sup ++}, Li{sup +}, AG{sup +}, Na{sup +}, Li, K{sup +}, Na, K. The isotopic discrimination appears to provide qualitative information about the strength of the cation-solvent interaction in liquid ammonia.