Over-expression in Escherichia coli of a thermally stable and regio-selective nitrile hydratase from Comamonas testosteroni 5-MGAM-4D

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• 作者：Kelly L. Petrillo ; Shijun Wu ; Eugenia C. Hann ; Frederick B. Cooling ; Arie Ben-Bassat ; John E. Gavagan ; Robert DiCosimo and Mark S. Payne
• 刊名：Applied Microbiology and Biotechnology
• 出版年：2005
• 出版时间：June 2005
• 年：2005
• 卷：67
• 期：5
• 页码：664-670
• 全文大小：388 KB

文摘

The genes encoding a thermally stable and regio-selective nitrile hydratase (NHase) and an amidase from Comamonas testosteroni 5-MGAM-4D have been cloned and sequenced, and active NHase has been over-produced in Escherichia coli. Maximal activity requires co-expression of a small open reading frame immediately downstream from the NHase beta subunit gene. Compared to the native organism, the E. coli biocatalyst has nearly threefold more NHase activity on a dry cell weight basis, and this activity is significantly more thermally stable. In addition, this biocatalyst converts a wide spectrum of nitrile substrates to the corresponding amides. Such versatility and robustness are desirable attributes of a biocatalyst intended for use in commercial applications.