Arginine deiminase mutant methods of using the same

Patent metadata

Field	Value
Publication number	US-10815470-B2
Application number	US-201916718743-A
Country	US
Kind code	B2
Filing date	Dec 18, 2019
Priority date	Mar 2, 2017
Publication date	Oct 27, 2020
Grant date	Oct 27, 2020

Abstract

An arginine deiminase mutant with improved enzyme activity and temperature stability and application thereof were provided, belonging to the technical field of genetic engineering and enzyme engineering. The arginine deiminase mutant is proline, namely Gly292 Pro, mutated from glycine near an enzyme active center. A wild-type arginine deiminase arcA coding gene is molecularly modified by a site-directed mutation technique to obtain a mutant enzyme ADIG292P, which has glycine at position 292 of an amino acid sequence of the wild type arginine deiminase mutated to proline. The arginine deiminase, modified by site-directed mutation, of the present invention has 1.5 times of increase in enzyme activity and 5.43 times of increase in half-life period at 40° C. compared with the wild-type enzyme, which solves the problems of low catalytic ability and temperature stability during the catalytic synthesis of citrulline using arginine deiminase, and lays a foundation for industrial production of efficient synthesis of citrulline and medication application.

First claim

What is claimed is:

1. A gene encoding an arginine deaminase mutant, wherein the nucleotide sequence of the gene is set forth in SEQ ID NO: 2, and wherein the arginine deaminase mutant comprises mutation Gly292Pro compared to a parent arginine deiminase, greater enzyme activity and greater temperature stability than the parent arginine deiminase.

Assignees

• Univ Jiangnan

Inventors

• Zhang Tao
• Jiang Bo
• Jiang Hangyu
• Mu Wanmeng
• Miao Ming

Classifications

• A61K38/50

  acting on carbon-nitrogen bonds, other than peptide bonds (3.5), e.g. asparaginase · CPC title

• C12P13/10

  Citrulline; Arginine; Ornithine · CPC title

• A61P35/02

  specific for leukemia · CPC title

• C12N2800/101

  for bacteria · CPC title

• C12N9/78 Primary

  acting on carbon to nitrogen bonds other than peptide bonds (3.5) · CPC title