Halohydrin dehalogenase mutant, engineering bacterium and application of halohydrin dehalogenase mutant

• Main Authors: ZHENG YUGUO, WAN XINYU, JEONG IN-JO, TANG XIAOLING
• Format: Patent
• Language: Chinese; English
• Published: 12.04.2022
• 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; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention discloses a halohydrin dehalogenase mutant, an engineering bacterium and application of the halohydrin dehalogenase mutant and the engineering bacterium. The halohydrin dehalogenase mutant is characterized in that the mutant is obtained by performing single mutation or multiple mutation on the 105th site, the 134 site, the 136th site, the 137th site, the 178th site or the 186th site of an amino acid sequence shown in SEQ ID NO.2. The e.e. Value of the product (S)-epichlorohydrin of the halohydrin dehalogenase mutant exceeds 99% by utilizing a water-organic two-phase buffer solution system (ethyl acetate: sodium phosphate buffer solution = 6: 4), the wet cell enzyme activity is respectively improved from 23.56 U/mL to 100-600 U/mL, and the yield reaches 15.9%-80.2%. 本发明公开了一种卤醇脱卤酶突变体、工程菌及其应用，其特征在于所述突变体是将SEQ ID NO.2所示氨基酸序列第105位、第134位、第136位、第137位、第178位或第186位进行单突变或多突变获得。本发明利用水-有机双相缓冲溶液体系(乙酸乙酯：磷酸钠缓冲液＝6:4)使得卤醇脱卤酶突变体的产物(S)-环氧氯丙烷的e.e.值超过99％，湿细胞酶活分别从23.56U/mL提高至100-600U/mL，产率达15.9％-80.2％。