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筛选、培养和生物催化性能强的 Rhodococcus boritolerans FW815 2,2-二甲基环丙烷甲腈水合酶活性。

Screening, cultivation, and biocatalytic performance of Rhodococcus boritolerans FW815 with strong 2,2-dimethylcyclopropanecarbonitrile hydratase activity.

机构信息

Institute of Bioengineering, Zhejiang University of Technology, Hangzhou, People's Republic of China.

出版信息

J Ind Microbiol Biotechnol. 2012 Mar;39(3):409-17. doi: 10.1007/s10295-011-1029-1. Epub 2011 Sep 4.

Abstract

In this work, a mild, efficient bioconversion of 2,2-dimethylcyclopropanecarbonitrile (DMCPCN) to 2,2-dimethylcyclopropanecarboxamide (DMCPCA) in distilled water system was developed. The isolate FW815 was screened using the enrichment culture technique, displaying strong DMCPCN hydratase activity, and was identified as Rhodococcus boritolerans based on morphological, physiological, biochemical tests and 16S rRNA gene sequencing. Cultivation outcomes indicated that R. boritolerans FW815 was a neutrophile, with a growth optimum of 28-32°C; its DMCPCN hydratase belonged to the Fe-type family, and was most active at 38-42°C, pH 7.0, with maximal activity of 4.51 × 10(4) U g(-1) DCW. R. boritolerans FW815 was found to be DMCPCA amidase-negative, eliminating the contamination of dimethylcyclopropanecarboxylic acid. Moreover, it displayed high activity and acceptable reusability in the non-buffered distilled water system, comparable to those in pH 7.0 phosphate buffer (50.0 mmol l(-1)).

摘要

在这项工作中,开发了一种在蒸馏水系中温和、高效的 2,2-二甲基环丙甲腈(DMCPCN)到 2,2-二甲基环丙甲酰胺(DMCPCA)的生物转化方法。通过富集培养技术筛选出的分离株 FW815 显示出很强的 DMCPCN 水合酶活性,根据形态学、生理学、生物化学试验和 16S rRNA 基因测序,鉴定为波罗的海分枝杆菌。培养结果表明,R. boritolerans FW815 是一种嗜中性菌,最适生长温度为 28-32°C;其 DMCPCN 水合酶属于 Fe 型家族,最适温度为 38-42°C,pH7.0,最大活性为 4.51×10(4)U g(-1) DCW。发现 R. boritolerans FW815 为 DMCPCA 酰胺酶阴性,消除了二甲基环丙羧酸的污染。此外,它在非缓冲蒸馏水系中表现出高活性和可接受的可重复使用性,与 pH7.0 磷酸盐缓冲液(50.0mmol l(-1))相当。

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