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Dehalogenase

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https://www.readbyqxmd.com/read/28641560/functional-and-catalytic-characterization-of-the-detoxifying-enzyme-haloalkane-dehalogenase-from-rhizobium-leguminosarum
#1
Nikolaos Georgakis, Evangelia Chronopoulou, Maria Anna Gad, Dimitrios Skliros, Rodica Efrose, Emmanouil Flemetakis, Nikolaos E Labrou
Haloalkanes are acutely toxic to animals and humans and their toxic effects are mainly observed in the liver, kidneys and central nervous system. In the present work, the haloalkane dehalogenase from Rhizobium leguminosarum bv. trifolii (DrlA) was characterized. Reverse transcription polymerase chain reaction analysis and enzyme activity assays revealed that the DrlA gene expression in R. leguminosarum bv. trifolii is induced by 1,2-dibromoethane (1,2-DBE) during the early exponential phase. The gene of the enzyme was isolated, cloned and expressed in E...
June 20, 2017: Protein and Peptide Letters
https://www.readbyqxmd.com/read/28637831/oshad1-a-haloacid-dehalogenase-like-apase-enhances-phosphate-accumulation
#2
Bipin Kumar Pandey, Poonam Mehra, Lokesh Verma, Jyoti Bhadouria, Jitender Giri
Phosphorus (P) deficiency limits plant growth and crop yield. Since, plants can absorb only inorganic form of P (Pi), a large portion of soil P (organic and inorganic P complexes) remains largely unused. Here, we identified and characterized a PHR2 regulated; novel low Pi responsive haloacid dehalogenase (HAD)-like hydrolase, OsHAD1. While, OsHAD1 is a functional HAD protein having both acid phosphatase and phytase activity; it showed little homology with other known low Pi responsive HAD superfamily members...
June 21, 2017: Plant Physiology
https://www.readbyqxmd.com/read/28631300/a-bacterial-chloroform-reductive-dehalogenase-purification-and-biochemical-characterization
#3
Bat-Erdene Jugder, Susanne Bohl, Helene Lebhar, Robert D Healey, Mike Manefield, Christopher P Marquis, Matthew Lee
We report herein the purification of a chloroform (CF)-reducing enzyme, TmrA, from the membrane fraction of a strict anaerobe Dehalobacter sp. strain UNSWDHB to apparent homogeneity with an approximate 23-fold increase in relative purity compared to crude lysate. The membrane fraction obtained by ultracentrifugation was solubilized in Triton X-100 in the presence of glycerol, followed by purification by anion exchange chromatography. The molecular mass of the purified TmrA was determined to be 44.5 kDa by SDS-PAGE and MALDI-TOF/TOF...
June 20, 2017: Microbial Biotechnology
https://www.readbyqxmd.com/read/28631290/a-h2-oxidizing-1-2-3-trichlorobenzene-reducing-multienzyme-complex-isolated-from-the-obligately-organohalide-respiring-bacterium-dehalococcoides-mccartyi-strain-cbdb1
#4
Stefanie Hartwig, Nadya Dragomirova, Anja Kublik, Dominique Türkowsky, Martin von Bergen, Ute Lechner, Lorenz Adrian, R Gary Sawers
Dehalococcoides mccartyi is a small, slow-growing bacterium of the phylum Chloroflexi that conserves energy using aliphatic and aromatic organohalides as electron acceptors, and hydrogen as sole electron donor. A recent study identified a protein complex in the membrane of strain CBDB1 comprising a Hup hydrogenase, a complex iron-sulphur molybdoprotein and a reductive dehalogenase (RdhA) that catalyses reduction of 1,2,3,4-tetrachlorobenzene. Using a combination of size-exclusion chromatography, in-gel hydrogenase activity-staining, immunological analysis and mass spectrometry, we identified here a large molecular mass protein complex solubilized from the cytoplasmic membrane of D...
June 19, 2017: Environmental Microbiology Reports
https://www.readbyqxmd.com/read/28618195/novel-reductive-dehalogenases-from-the-marine-sponge-associated-bacterium-desulfoluna-spongiiphila
#5
Jie Liu, Nora Lopez, Young-Beom Ahn, Tatyana Goldberg, Yana Bromberg, Lee J Kerkhof, Max M Häggblom
Desulfoluna spongiiphila strain AA1 is an organohalide respiring bacterium, isolated from the marine sponge Aplysina aerophoba, that can use brominated and iodinated phenols, in addition to sulfate and thiosulfate as terminal electron acceptors. The genome of Desulfoluna spongiiphila strain AA1 is approximately 6.5 Mb. Three putative reductive dehalogenase (rdhA) genes involved in respiratory metabolism of organohalides were identified within the sequence. Conserved motifs found in respiratory reductive dehalogenases (a twin arginine translocation signal sequence and two iron-sulfur clusters) were present in all three putative AA1 rdhA genes...
June 15, 2017: Environmental Microbiology Reports
https://www.readbyqxmd.com/read/28618081/loss-of-the-ssra-genome-island-led-to-partial-debromination-in-the-pbde-respiring-dehalococcoides-mccartyi-strain-gy50
#6
Chang Ding, Matthew J Rogers, Kun-Lin Yang, Jianzhong He
Polybrominated diphenyl ethers (PBDEs), chemicals commonly used as flame-retardants in consumer products, are emerging persistent organic pollutants that are ubiquitous in the environment. In this study, we report the PBDE-respiring isolate - Dehalococcoides mccartyi strain GY50, which debrominates the most toxic tetra- and penta-BDE congeners (∼1.4 µM) to diphenyl ether within 12 days with hydrogen as the electron donor. The complete genome sequence revealed 26 reductive dehalogenase homologous genes (rdhAs), among which three genes (pbrA1, pbrA2, and pbrA3) were highly expressed during PBDE debromination...
June 15, 2017: Environmental Microbiology
https://www.readbyqxmd.com/read/28593475/functional-metagenomics-of-oil-impacted-mangrove-sediments-reveals-high-abundance-of-hydrolases-of-biotechnological-interest
#7
Júlia Ronzella Ottoni, Lucélia Cabral, Sanderson Tarciso Pereira de Sousa, Gileno Vieira Lacerda Júnior, Daniela Ferreira Domingos, Fábio Lino Soares Junior, Mylenne Calciolari Pinheiro da Silva, Joelma Marcon, Armando Cavalcante Franco Dias, Itamar Soares de Melo, Anete Pereira de Souza, Fernando Dini Andreote, Valéria Maia de Oliveira
Mangroves are located in coastal wetlands and are susceptible to the consequences of oil spills, what may threaten the diversity of microorganisms responsible for the nutrient cycling and the consequent ecosystem functioning. Previous reports show that high concentration of oil favors the incidence of epoxide hydrolases and haloalkane dehalogenases in mangroves. This finding has guided the goals of this study in an attempt to broaden the analysis to other hydrolases and thereby verify whether oil contamination interferes with the prevalence of particular hydrolases and their assigned microorganisms...
July 2017: World Journal of Microbiology & Biotechnology
https://www.readbyqxmd.com/read/28587829/pegylation-with-the-thiosuccinimido-butylamine-linker-significantly-increases-the-stability-of-haloalkane-dehalogenase-dhaa
#8
Yuan-Zhong Zhao, Wei-Li Yu, He Zheng, Xuan Guo, Nan Guo, Tao Hu, Jin-Yi Zhong
Haloalkane dehalogenase (HLD) can catalyze the hydrolytic dehalogenation of halogenated compounds. However, HLD suffers from the poor stability to resist the environmental stress. PEGylation is an effective approach to enhance the stability of enzymes. The linker is an important stabilization factor of PEGylation. Thus, the linkers of the PEGylated HLD were optimized to improve the stability of HLD in the present study. The PEGylated haloalkane dehalogenase DhaAs with methylamine (Ml), carbamate (Cm) and thiosuccinimido butylamine (Tb) linkers were prepared, respectively...
June 3, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28533514/isolation-and-genomic-characterization-of-a-dehalococcoides-strain-suggests-genomic-rearrangement-during-culture
#9
Masafumi Yohda, Kentaro Ikegami, Yuto Aita, Mizuki Kitajima, Ayane Takechi, Megumi Iwamoto, Tomomi Fukuda, Noriyoshi Tamura, Junji Shibasaki, Seiji Koike, Daisuke Komatsu, Sakari Miyagi, Minoru Nishimura, Yoshihito Uchino, Akino Shiroma, Makiko Shimoji, Hinako Tamotsu, Noriko Ashimine, Misuzu Shinzato, Shun Ohki, Kazuma Nakano, Kuniko Teruya, Kazuhito Satou, Takashi Hirano, Osami Yagi
We have developed and characterized a bacterial consortium that reductively dechlorinates trichloroethene to ethene. Quantitative PCR analysis for the 16S rRNA and reductive dehalogenase genes showed that the consortium is highly enriched with Dehalococcoides spp. that have two vinyl chloride reductive dehalogenase genes, bvcA and vcrA, and a trichloroethene reductive dehalogenase gene, tceA. The metagenome analysis of the consortium by the next generation sequencer SOLiD 3 Plus suggests that a Dehalococcoides sp...
May 22, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28527381/enantioselectivity-of-haloalkane-dehalogenase-linb-on-the-degradation-of-1-2-dichloropropane-a-qm-mm-study
#10
Xiaowen Tang, Ruiming Zhang, Yanwei Li, Qingzhu Zhang, Wenxing Wang
The hydrolysis dechlorination mechanism of a chiral organochlorinepollutant, 1,2-dichloropropane (DCP), catalyzed by haloalkane dehalogenase LinB has been investigated by using a combined quantum mechanics/molecular mechanics method. LinB was confirmed to be enantioselective towards the catabolism of the racemic mixture. Based on the SN2 nucleophilic substitution mechanism, the dechlorination process was identified as the rate-determining step in LinB-catalyzed degradation of 1,2-dichloropropane, the Boltzmann-weighted average potential barrier of which is 18...
May 12, 2017: Bioorganic Chemistry
https://www.readbyqxmd.com/read/28499743/resolution-of-the-uncertainty-in-the-kinetic-mechanism-for-the-trans-3-chloroacrylic-acid-dehalogenase-catalyzed-reaction
#11
Jamison P Huddleston, Susan C Wang, Kenneth A Johnson, Christian P Whitman
trans- and cis-3-Chloroacrylic acid dehalogenase (CaaD and cis-CaaD, respectively) catalyze the hydrolytic dehalogenation of their respective isomers and represent key steps in the bacterial conversion of 1,3-dichloropropene to acetaldehyde. In prior work, a kinetic mechanism for the CaaD-catalyzed reaction could not be unequivocally determined because (1) the order of product release could not be determined and (2) the fluorescence factor for the enzyme species, E*PQ (where P = bromide and Q = malonate semialdehyde, the two products of the reaction) could not be assigned...
June 1, 2017: Archives of Biochemistry and Biophysics
https://www.readbyqxmd.com/read/28424844/microbial-reductive-dehalogenation-of-trihalomethanes-by-a-dehalobacter-containing-co-culture
#12
Siyan Zhao, Matthew J Rogers, Jianzhong He
Trihalomethanes such as chloroform and bromoform, although well-known as a prominent class of disinfection by-products, are ubiquitously distributed in the environment due to widespread industrial usage in the past decades. Chloroform and bromoform are particularly concerning, of high concentrations detected and with long half-lives up to several hundred days in soils and groundwater. In this study, we report a Dehalobacter- and Desulfovibrio-containing co-culture that exhibits dehalogenation of chloroform (~0...
April 19, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28421054/isolation-and-characterization-of-dehalobacter-sp-strain-tecb1-including-identification-of-tcba-a-novel-tetra-and-trichlorobenzene-reductive-dehalogenase
#13
Ricardo Alfán-Guzmán, Haluk Ertan, Mike Manefield, Matthew Lee
Dehalobacter sp. strain TeCB1 was isolated from groundwater near Sydney, Australia, that is polluted with a range of organochlorines. The isolated strain is able to grow by reductive dechlorination of 1,2,4,5-tetrachlorobenzene to 1,3- and 1,4-dichlorobenzene with 1,2,4-trichlorobenzene being the intermediate daughter product. Transient production of 1,2-dichlorobenzene was detected with subsequent conversion to monochlorobenzene. The dehalogenation capability of strain TeCB1 to respire 23 alternative organochlorines was examined and shown to be limited to the use of 1,2,4,5-tetrachlorobenzene and 1,2,4-trichlorobenzene...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28419658/ancestral-haloalkane-dehalogenases-show-robustness-and-unique-substrate-specificity
#14
Petra Babkova, Eva Sebestova, Jan Brezovsky, Radka Chaloupkova, Jiri Damborsky
Ancestral sequence reconstruction (ASR) represents a powerful approach for empirical testing structure-function relationships of diverse proteins. We employed ASR to predict sequences of five ancestral haloalkane dehalogenases (HLDs) from the HLD-II subfamily. Genes encoding the inferred ancestral sequences were synthesized and expressed in Escherichia coli and the resurrected ancestral enzymes AncHLD1-5 were experimentally characterized. Strikingly, the ancestral HLDs exhibited significantly enhanced thermodynamic stability compared to extant enzymes (ΔTm up to 24 °C)...
April 17, 2017: Chembiochem: a European Journal of Chemical Biology
https://www.readbyqxmd.com/read/28411224/concurrent-haloalkanoate-degradation-and-chlorate-reduction-by-pseudomonas-chloritidismutans-aw-1-t
#15
Peng Peng, Ying Zheng, Jasper J Koehorst, Peter J Schaap, Alfons J M Stams, Hauke Smidt, Siavash Atashgahi
Haloalkanoates are environmental pollutants that can be degraded aerobically by microorganisms producing hydrolytic dehalogenases. However, there is lack of information about anaerobic degradation of haloalkanoates. Genome analysis of Pseudomonas chloritidismutans AW-1(T), a facultative anaerobic chlorate-reducing bacterium, showed presence of two putative haloacid dehalogenase genes, l-dex and dehI, encoding an L-2-haloacid dehalogenase (L-DEX) and a halocarboxylic acid dehydrogenase (DehI). Hence, we studied concurrent degradation of haloalkanoates and chlorate as a yet unexplored trait of strain AW-1(T) The deduced amino acid sequences of L-DEX and DehI revealed 33-37% and 26-86% identities with biochemically/structurally characterized L-DEX and D-, DL-2-haloacid dehalogenase enzymes, respectively...
April 14, 2017: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/28397170/the-organohalide-respiring-bacterium-sulfurospirillum-multivorans-a-natural-source-for-unusual-cobamides
#16
REVIEW
Torsten Schubert
Cobamides ('complete' corrinoids) are essential for organohalide-respiring bacteria because they act as cofactors of reductive dehalogenases (RDases). RDases are the key enzymes in organohalide respiration, a process relevant for environmental remediation. More than a decade ago, the unusual norpseudo-B12 was identified as cofactor of the tetrachloroethene RDase (PceA) purified from the epsilonproteobacterium Sulfurospirillum multivorans. Since then, the question was raised whether or not the production of the uncommon cobamide is a specific adaptation to the requirements of PceA...
May 2017: World Journal of Microbiology & Biotechnology
https://www.readbyqxmd.com/read/28381628/a-puzzling-homology-a-brittle-star-using-a-putative-cnidarian-type-luciferase-for-bioluminescence
#17
Jérôme Delroisse, Esther Ullrich-Lüter, Stefanie Blaue, Olga Ortega-Martinez, Igor Eeckhaut, Patrick Flammang, Jérôme Mallefet
Bioluminescence relies on the oxidation of a luciferin substrate catalysed by a luciferase enzyme. Luciferins and luciferases are generic terms used to describe a large variety of substrates and enzymes. Whereas luciferins can be shared by phylogenetically distant organisms which feed on organisms producing them, luciferases have been thought to be lineage-specific enzymes. Numerous light emission systems would then have co-emerged independently along the tree of life resulting in a plethora of non-homologous luciferases...
April 2017: Open Biology
https://www.readbyqxmd.com/read/28359974/purifying-cloning-and-characterizing-a-novel-dehalogenase-from-bacillus-sp-gzt-to-enhance-the-biodegradation-of-2-4-6-tribromophenol-in-water
#18
Zhishu Liang, Guiying Li, Taicheng An
2,4,6-Tribromophenol (TBP), an intermediate of brominated flame retardants, can easily release to environment and recalcitrant to degradation. Previously, Bacillus sp. GZT, a pure aerobic strain capable of simultaneously debrominating and mineralizing TBP, was successfully isolated by us. To further obtain a practical application and dig up its TBP degradation mechanism, a total of 46.7-fold purification of a novel dehalogenase with a final specific activity of 18.9 U mg(-1) and a molecular mass of 63.4 kDa was achieved...
March 27, 2017: Environmental Pollution
https://www.readbyqxmd.com/read/28351914/the-n-acetylmuramic-acid-6-phosphate-phosphatase-mupp-completes-the-pseudomonas-peptidoglycan-recycling-pathway-leading-to-intrinsic-fosfomycin-resistance
#19
Marina Borisova, Jonathan Gisin, Christoph Mayer
Bacterial cells are encased in and stabilized by a netlike peptidoglycan (PGN) cell wall that undergoes turnover during bacterial growth. PGN turnover fragments are frequently salvaged by the cells via a pathway referred to as PGN recycling. Two different routes for the recycling of the cell wall sugar N-acetylmuramic acid (MurNAc) have been recognized in bacteria. In Escherichia coli and related enterobacteria, as well as in most Gram-positive bacteria, MurNAc is recovered via a catabolic route requiring a MurNAc 6-phosphate etherase (MurQ in E...
March 28, 2017: MBio
https://www.readbyqxmd.com/read/28334478/different-structural-origins-of-the-enantioselectivity-of-haloalkane-dehalogenases-toward-linear-%C3%AE-haloalkanes-open-solvated-versus-occluded-desolvated-active-sites
#20
Veronika Liskova, Veronika Stepankova, David Bednar, Jan Brezovsky, Zbynek Prokop, Radka Chaloupkova, Jiri Damborsky
The enzymatic enantiodiscrimination of linear β-haloalkanes is difficult because the simple structures of the substrates prevent directional interactions. Herein we describe two distinct molecular mechanisms for the enantiodiscrimination of the β-haloalkane 2-bromopentane by haloalkane dehalogenases. Highly enantioselective DbjA has an open, solvent-accessible active site, whereas the engineered enzyme DhaA31 has an occluded and less solvated cavity but shows similar enantioselectivity. The enantioselectivity of DhaA31 arises from steric hindrance imposed by two specific substitutions rather than hydration as in DbjA...
March 23, 2017: Angewandte Chemie
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