論文 - 詳細
| RRC ID | 17467 |
|---|---|
| 著者 | Kato D, Yoshida H, Takeo M, Negoro S, Ohta H. |
| タイトル | Purification and gene cloning of an enantioselective thioesterification enzyme from Brevibacterium ketoglutamicum KU1073, a deracemization bacterium of 2-(4-chlorophenoxy)propanoic acid. |
| ジャーナル | Biosci Biotechnol Biochem |
| Abstract |
We succeeded in the purification and gene cloning of a new enzyme, α-methyl carboxylic acid deracemizing enzyme 1 (MCAD1) from Brevibacterium ketoglutamicum KU1073, which catalyzes the (S)-enantioselective thioesterification reaction of 2-(4-chlorophenoxy)propanoic acid (CPPA). The cloned gene of MCAD1 contained an ORF of 1,623 bp, encoding a polypeptide of 540 amino acids. In combination with cofactors ATP, coenzyme A (CoASH), and Mg(2+), MCAD1 demonstrated perfect enantioselectivity toward CPPA. The optimal pH and temperature for reaction were found to be 7.25 and 30 °C. Under these conditions, the K(m) and k(cat) values for (S)-CPPA were 0.92 ± 0.17 mM and 0.28 ± 0.026 s(-1) respectively. The results for substrate specificity revealed that MCAD1 had highest activity toward fatty acid tails with a medium chain-length (C(8)). This result indicates that MCAD1 should be classified into a family of medium-chain acyl-CoA synthetase. This novel activity has never been reported for this family. |
| 巻・号 | 74(12) |
| ページ | 2405-12 |
| 公開日 | 2010-1-1 |
| DOI | 10.1271/bbb.100431 |
| PII | JST.JSTAGE/bbb/100431 |
| PMID | 21150124 |
| MeSH | 2-Methyl-4-chlorophenoxyacetic Acid / analogs & derivatives* 2-Methyl-4-chlorophenoxyacetic Acid / chemistry 2-Methyl-4-chlorophenoxyacetic Acid / metabolism Amino Acid Sequence Brevibacterium / enzymology* Brevibacterium / genetics Brevibacterium / metabolism Cell-Free System / enzymology Cloning, Molecular Coenzyme A Ligases / chemistry Coenzyme A Ligases / genetics* Coenzyme A Ligases / isolation & purification* Coenzyme A Ligases / metabolism Enzyme Stability Esterification Molecular Sequence Data Phenylpropionates / chemistry* Phenylpropionates / metabolism* Recombinant Proteins / chemistry Recombinant Proteins / genetics Recombinant Proteins / isolation & purification Recombinant Proteins / metabolism Sequence Analysis, DNA Stereoisomerism Substrate Specificity |
| IF | 1.516 |
| 引用数 | 3 |
| WOS 分野 | FOOD SCIENCE & TECHNOLOGY CHEMISTRY, APPLIED BIOTECHNOLOGY & APPLIED MICROBIOLOGY BIOCHEMISTRY & MOLECULAR BIOLOGY |
| リソース情報 | |
| 一般微生物 | JCM 3208 |