Cyclodextrin glucosyltransferase (CGTase) with improved organic solvent tolerance and preparation method of cyclodextrin glucosyltransferase

• Main Authors: FAN CHAOYUE, NI YE, HAN RUIZHI, YI LINGFENG, JIANG YULIN
• Format: Patent
• Language: Chinese; English
• Published: 21.12.2021
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention discloses cyclodextrin glucosyltransferase with improved organic solvent tolerance and a preparation method of the cyclodextrin glucosyltransferase (CGTase), and belongs to the technical field of enzyme engineering and genetic engineering. Two CGTase mutants with improved organic solvent (dimethyl sulfoxide and ethanol) tolerance are constructed. Taking genistein as an example, the two mutants are applied to glycosylation reaction of genistein, and the result shows that the specificity of long-chain glycosylation products generated by T599H/N600S/Y601G and T599D/N600D/Y601G is obviously superior to that of a wild type WT, and is 2.1 times and 2.5 times of that of the wild type WT respectively. The two CGTase mutants with improved organic solvent tolerance are obtained, the enzymatic efficiency in a glycosyl transferase organic reaction system is promoted, the industrial application range of CGTase is expanded, and the CGTase has a great application prospect. 本发明公开了一种有机溶剂耐受性提高的环糊精葡萄糖基转移酶及其制备方法，属于