3-ketosteroid-delta1-dehydrogenase mutant, coding gene, vector and application

• Main Authors: LIU ZHIQIANG, ZHOU DENGFENG, HUANG LIANGGANG, ZHENG YUGUO, ZHANG BO, NI YEWEN, ZHOU JUNPING
• Format: Patent
• Language: Chinese; English
• Published: 01.03.2022
• 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; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention discloses a 3-ketosteroid-delta1-dehydrogenase mutant, a coding gene, a vector and application, and belongs to the technical field of enzyme engineering. The 3-ketosteroid-delta1-dehydrogenase mutant disclosed by the invention is obtained by site-directed mutagenesis of the 214th site and/or the 499th site of the 3-ketosteroid-delta1-dehydrogenase mutant of which the amino acid sequence is shown as SEQ ID NO.1. The invention further discloses a preparation method of the 3-ketosteroid-delta1-dehydrogenase mutant. According to the invention, site-directed mutagenesis is carried out through multi-sequence amino acid comparison, so that the catalytic efficiency of the 3-ketosteroid-delta1-dehydrogenase is improved, and the yield of the 3-ketosteroid-delta1-dehydrogenase is further improved. The recombinant escherichia coli of the 3-ketosteroid-delta1-dehydrogenase constructed by the invention can improve the enzyme activity of the 3-ketosteroid-delta1-dehydrogenase by 1.5 times compared with the ori