Malate dehydrogenase mutant gene and construction method and application thereof

• Main Authors: GE YADONG, SU FENGZHI, JIANG LULU, HOU SHAOLIN
• Format: Patent
• Language: Chinese; English
• Published: 05.02.2019
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMES; 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 malate dehydrogenase mutant gene and a construction method and application thereof, and belongs to the technical field of biological genetic engineering. The malate dehydrogenase mutant gene is characterized in that streptomyces coelicolor malate dehydrogenase is taken as a template, a gene sequence capable of expressing the malate dehydrogenase is constructed through genetic engineering measures such as primer design, stepwise PCR (polymerase chain reaction), fusion PCR, transformed bacteria resistance screening and nucleotide sequencing, and the constructed malate dehydrogenase is transferred into DE3 strains of escherichia coli to be expressed; the activity of the expressed malate dehydrogenase is two times or above that of malate dehydrogenase under wild-typesample expression, and more efficient resolution and purification enzymes are provided for production of D-malic acid. 本发明公开了种苹果酸脱氢酶突变基因、构建方法及其应用，属于生物基因工程技术领域，突变苹果酸脱氢酶基因以天蓝色链霉菌苹果酸脱氢酶为模板通过引物设计，分步PCR，融合PCR，转化菌抗性筛选，核苷酸测序的基因工程的手段