Purification and Characterization of Catechol 2,3-Dioxygenase from the Aniline Degradation Pathway of Acinetobacter sp. YAA and Its Mutant Enzyme, Which Resists Substrate Inhibition
スポンサーリンク
概要
- 論文の詳細を見る
Catechol 2,3-dioxygenase (C23O), a key enzyme in the meta-cleavage pathway of catechol metabolism, was purified from cell extract of recombinant Escherichia coli JM109 harboring the C23O gene (atdB) cloned from an aniline-degrading bacterium Acinetobacter sp. YAA. SDS–polyacrylamide gel electrophoresis and gel filtration chromatography analysis suggested that the enzyme (AtdB) has a molecular mass of 35 kDa as a monomer and forms a tetrameric structure. It showed relative meta-cleavage activities for the following catechols tested: catechol (100%), 3-methylcatechol (19%), 4-methylcatechol (57%), 4-chlorocatechol (46%), and 2,3-dihydroxybiphenyl (5%). To elevate the activity, a DNA self-shuffling experiment was carried out using the atdB gene. One mutant enzyme, named AtdBE286K, was obtained. It had one amino acid substitution, E286K, and showed 2.4-fold higher C23O activity than the wild-type enzyme at 100 μM. Kinetic analysis of these enzymes revealed that the wild-type enzyme suffered from substrate inhibition at >2 μM, while the mutant enzyme loosened substrate inhibition.
- 社団法人 日本農芸化学会の論文
著者
-
Takeo Masahiro
Department of Materials Science and Chemistry, Graduate School of Engineering, University of Hyogo
-
Negoro Seiji
Department of Materials Science and Chemistry, Graduate School of Engineering, University of Hyogo
-
Nishimura Munehiro
Department of Materials Science and Chemistry, Graduate School of Engineering, University of Hyogo
-
Takahashi Hana
Department of Materials Science and Chemistry, Graduate School of Engineering, University of Hyogo
-
Takeo Masahiro
兵庫県立大学 工学研究科物質系工学専攻
-
SHIRAI Mizuho
Department of Materials Science and Chemistry, Graduate School of Engineering, University of Hyogo
関連論文
- Enzyme-linked Immunosorbent Assay for Nonylphenol Using Antibody-Bound Microfluid Filters in Vertical Fluidic Operation(METHODS)
- Immunoassay Using Microfluid Filters Constructed by Deep X-Ray Lithography
- Novel Characteristics of Multifunctional Fluid Filters Fabricated by High-Energy Synchrotron Radiation Lithography
- A New Micro-Chemical Reactor Using Fluid Filters for Vertical Fluid Flow Operation
- Mutational Analysis of the Metabolism of 2,6-Naphthalenedisulfonate by Pigmentiphaga sp. NDS-2
- フロック形成菌Pigmentiphaga sp. NDS-1による2,6-ナフタレンジスルホン酸の生物分解
- Characterization of Hybrid Enzymes between 6-Aminohexanoate-Dimer Hydrolase and Its Analogous Protein
- Alteration of Catalytic Function of 6-Aminohexanoate-Dimer Hydrolase by Site-Directed Mutagenesis
- Molecular Cloning and Sequencing of the Phenol Hydroxylase Gene from Pseudomonas putida BH
- Expression of the Xylan-Degrading Genes of Bacillus pumilus IPO in Saccharomyces cerevisiae
- 1P042 Molecular dynamics studies on Mutational structures of a Nylon-6 Byproduct-degrading Enzyme(Protein:Structure & Function,The 48th Annual Meeting of the Biophysical Society of Japan)
- Construction and Characterization of N-Terminally Truncated DNA Polymerase from Thermus thermophilus
- Analysis of a Promoter-Like Region in Flavobacterium Which Controls 6-Aminohexanoic Acid Linear Oligomer Hydrolase Gene Expression in Escherichia coli
- Trichloroethylene Degradation by Genetically Engineered Bacteria Carrying Cloned Phenol Catabolic Genes
- Genetic Organization of Nylon-Oligomer-Degrading Enzymes from Alkalophilic Bacterium, Agromyces sp. KY5R(GENETICS, MOLECULAR BIOLOGY, AND GENE ENGINEERING)
- Purification and Characterization of Alkylcatechol 2,3-Dioxygenase from Butylphenol Degradation Pathway of Pseudomonas putida MT4(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)
- Characterization of Alkylphenol Degradation Gene Cluster in Pseudomonas putida MT4 and Evidence of Oxidation of Alkylphenols and Alkylcatechols with Medium-Length Alkyl Chain(ENVIRONMENTAL BIOTECHNOLOGY)
- Characterization of Endo-Type 6-Aminohexanoate-Oligomer Hydrolase from Flavobacterium sp.
- Structure of Isozyme Genes of Glucose Dehydrogenase from Bacillus megaterium IAM1030
- Purification and Characterization of a New Glucose Dehydrogenase from Vegetative Cells of Bacillus megaterium
- Simultaneous Degradation of 4-Nitrophenol and Picric Acid by Two Different Mechanisms of Rhodococcus sp. PN1
- Cloning and Characterization of a 4-Nitrophenol Hydroxylase Gene Cluster from Rhodococcus sp. PN1
- Plasmid-Coded Degradation of Salicylate Using the Catechol Cleavage Pathway of the Host
- Purification and Gene Cloning of an Enantioselective Thioesterification Enzyme from Brevibacterium ketoglutamicum KU1073, a Deracemization Bacterium of 2-(4-Chlorophenoxy)propanoic Acid
- Purification and Characterization of Catechol 2,3-Dioxygenase from the Aniline Degradation Pathway of Acinetobacter sp. YAA and Its Mutant Enzyme, Which Resists Substrate Inhibition
- Mineralization and Desulfonation of 3-Nitrobenzenesulfonic Acid by Alcaligenes sp. GA-1
- Biodegradability of 3-Methyl-4-Nitrophenol by 4-Nitrophenol-Degrading Bacteria
- Purification and Gene Cloning of an Enantioselective Thioesterification Enzyme from Brevibacterium ketoglutamicum KU1073, a Deracemization Bacterium of 2-(4-Chlorophenoxy)propanoic Acid
- Immunoassay Using Microfluid Filters Constructed by Deep X-Ray Lithography
- Purification and Characterization of Catechol 2,3-Dioxygenase from the Aniline Degradation Pathway of Acinetobacter sp. YAA and Its Mutant Enzyme, Which Resists Substrate Inhibition
- Identification and characterization of another 4-nitrophenol degradation gene cluster, nps, in Rhodococcus sp. strain PN1(ENVIRONMENTAL BIOTECHNOLOGY)
- Comparison of Acyl-CoA Synthetic Activities and Enantioselectivity toward 2-Arylpropanoic Acids in Firefly Luciferases