Biochemical characterization and substrate profiling of a new NADH-dependent enoate reductase from Lactobacillus casei
文献类型：期刊论文
| 作者 | Gao, Xiuzhen; Ren, Jie; Wu, Qiaqing; Zhu, Dunming |
| 刊名 | ENZYME AND MICROBIAL TECHNOLOGY
 |
| 出版日期 | 2012-06-10 |
| 卷号 | 51期号:1页码:26-34 |
| 关键词 | Carbon-carbon double bond Enoate reductase Bioreduction Biocatalysis |
| 英文摘要 | Carbon-carbon double bond of alpha,beta-unsaturated carbonyl compounds can be reduced by enoate reductase (ER), which is an important reaction in fine chemical synthesis. A putative enoate reductase gene from Lactobacillus casei str. Zhang was cloned into pET-21a(+) and expressed in Escherichia coli BL21 (DE3) host cells. The encoded enzyme (LacER) was purified by ammonium sulfate precipitation and treatment in an acidic buffer. This enzyme was identified as a NADH-dependent enoate reductase, which had a K-m of 0.034 +/- 0.006 mM and kcat of (3.2 +/- 0.2) x 10(3) s(-1) toward NADH using 2-cyclohexen-1-one as the substrate. Its K-m and k(cat) toward substrate 2-cyclohexen-1-one were 1.94 +/- 0.04 mM and (8.4 +/- 0.2) x 10(3) s(-1), respectively. The enzyme showed a maximum activity at pH 8.0-9.0. The optimum temperature of the enzyme was 50-55 degrees C, and LacER was relatively stable below 60 degrees C. The enzyme was active toward aliphatic alkenyl aldehyde, ketones and some cyclic anhydrides. Substituted groups of cyclic alpha,beta-unsaturated ketones and its ring size have positive or negative effects on activity. (R)-(-)-Carvone was reduced to (2R,5R)-dihydrocarvone with 99% conversion and 98% (diasteromeric excess: de) stereoselectivity, indicating a high synthetic potential of LacER in asymmetric synthesis. (C) 2012 Elsevier Inc. All rights reserved. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 类目[WOS] | Biotechnology & Applied Microbiology |
| 研究领域[WOS] | Biotechnology & Applied Microbiology |
| 关键词[WOS] | OLD-YELLOW-ENZYME ; PENTAERYTHRITOL TETRANITRATE REDUCTASE ; ASYMMETRIC REDUCTION ; TRANSFER HYDROGENATION ; CANDIDA-MACEDONIENSIS ; BETA(2)-AMINO ACIDS ; FAMILY ; SEQUENCE ; SPECIFICITY ; DISTINCT |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000307624900004 |
| 源URL | [http://124.16.173.210/handle/834782/1321]  |
| 专题 | 天津工业生物技术研究所_生物催化与绿色化工 朱敦明_期刊论文 |
| 作者单位 | Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Natl Engn Lab Ind Enzymes, Tianjin Airport Econ Area, Tianjin 300308, Peoples R China |
| 推荐引用方式 GB/T 7714 | Gao, Xiuzhen,Ren, Jie,Wu, Qiaqing,et al. Biochemical characterization and substrate profiling of a new NADH-dependent enoate reductase from Lactobacillus casei[J]. ENZYME AND MICROBIAL TECHNOLOGY,2012,51(1):26-34. |
| APA | Gao, Xiuzhen,Ren, Jie,Wu, Qiaqing,&Zhu, Dunming.(2012).Biochemical characterization and substrate profiling of a new NADH-dependent enoate reductase from Lactobacillus casei.ENZYME AND MICROBIAL TECHNOLOGY,51(1),26-34. |
| MLA | Gao, Xiuzhen,et al."Biochemical characterization and substrate profiling of a new NADH-dependent enoate reductase from Lactobacillus casei".ENZYME AND MICROBIAL TECHNOLOGY 51.1(2012):26-34. |
入库方式： OAI收割
来源：天津工业生物技术研究所
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。