Purification and Gene Cloning of an Enantioselective Thioesterification Enzyme from<i>Brevibacterium ketoglutamicum</i>KU1073, a Deracemization Bacterium of 2-(4-Chlorophenoxy)propanoic Acid
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- KATO Dai-ichiro
- Graduate School of Engineering, University of Hyogo
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- YOSHIDA Hiromitsu
- Graduate School of Engineering, University of Hyogo
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- TAKEO Masahiro
- Graduate School of Engineering, University of Hyogo
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- NEGORO Seiji
- Graduate School of Engineering, University of Hyogo
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- OHTA Hiromichi
- Department of Biosciences and Informatics, Keio University
書誌事項
- タイトル別名
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- Purification and Gene Cloning of an Enantioselective Thioesterification Enzyme from Brevibacterium ketoglutamicum KU1073, a Deracemization Bacterium of 2-(4-Chlorophenoxy)propanoic Acid
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説明
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 Mg2+, 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 Km and kcat 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 (C8). 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.
収録刊行物
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- Bioscience, Biotechnology, and Biochemistry
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Bioscience, Biotechnology, and Biochemistry 74 (12), 2405-2412, 2010
公益社団法人 日本農芸化学会
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詳細情報 詳細情報について
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- CRID
- 1390282681455997824
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- NII論文ID
- 10027727906
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- NII書誌ID
- AA10824164
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- ISSN
- 13476947
- 09168451
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- NDL書誌ID
- 10928495
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- 本文言語コード
- en
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- データソース種別
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- JaLC
- NDL
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