Zhejiang University

Zhejiang Chiral Medicine Chemicals Co. Ltd.

Renchao Zheng

Qin Zhang

Yuguo Zheng

Youming Huang

Tianchun Liu

Weirong Fan

Jianfeng Weng

Xiaoling Tang

Xiafeng Lu

Zheming Wu

The present invention discloses a nitrilase mutant and its construction method and its application in the synthesis of chiral intermediate of pregabalin in the technical field of bioengineering. The present invention, respectively, takes turnip nitrilase BrNIT and arabidopsis nitrilase AtNIT as parent, using peptide fragment displacement method, displaces the sites 226-286 of BrNIT amino acid sequence and sites 225-285 of AtNIT amino acid sequence with sites 225-285 ofL. nitrilase AaNIT, obtain nitrilase mutants BrNITand AtNITof which the amino acid sequence is as shown in SEQ ID NO.1 or SEQ ID NO.3. Compared with wild type nitrilase, the activity of the nitrilase mutant provided by the present invention in catalyzing and hydrolyzing racemic IBSN and the stereoselectivity of the product show substantial improvement, it can satisfy the requirements of industrial application, and has good application prospect in efficient catalysis of racemic IBSN to synthesize 3-cyano-5-methylhexanoic Acid.

Nitrilase mutant, construction method therefor, and application thereof

The disclosure provides a nitrilase from, which belongs to genus, family brassicaceae. The disclosure further provides the encoding gene, vector, recombinant bacterial strain, and the application in the manufacturing of (S)-3-cyano-5-methylhexanoic acid. The wet resting cells containing nitrilase Aa-Nit can kinetically resolve racemic IBSN at 1.2 M with a 42% conversion rate in 15 hr and &#x3e;99% ee value. The disclosure provides a regio- and stereoselective method for the preparation of (S)-3-cyano-5-methylhexanoic acid. This method provides an atom economical, mild, environmental friendly industrial method to manufacture (S)-3-cyano-5-methylhexanoic acid.

Nitrilase from arabis alpina, its encoding gene, vector, recombinant bacterial strain and uses thereof

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Growing community of inventors

Hangzhou, China

Renchao Zheng