Deinococcus radiodurans-derived P450 monooxygenase mutant and application thereof

• Main Authors: WAN NANWEI, CHEN YONGZHENG, DENG GUOZHONG, CUI HAIBO
• Format: Patent
• Language: Chinese; English
• Published: 10.01.2020
• 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 provides a deinococcus radiodurans-derived P450 monooxygenase mutant and application thereof in synthesis of (S)-2-chloro-1-phenylethanol chiral beta-halohydrin compounds. The P450 monooxygenase is subjected to several rounds of directed transformation to obtain P450 monooxygenase mutants N190D, N190F and N190F/V356L/A486E with improved asymmetric hydroxylation activity, wherein thecatalytic activity of the P450 monooxygenase mutant N190F/V356L/A486E is increased by 8.5 times that of wild P450 monooxygenase. The whole cell of the N190F/V356L/A486E mutant can be applied to catalysis of a benzylic asymmetric hydroxylation reaction of 1-chloro-2-phenylethane and derivatives thereof, the corresponding (S)-2-chloro-1-phenylethanol chiral beta-halohydrin compounds having the yieldof 88 percent and the optical purity of 98 percent ee can be obtained. 源于抗辐射奇异球菌P450单加氧酶突变体以及突变体合成(S)-2-氯-1-苯乙醇类手性β-卤代醇化合物的应用。本发明通过对P450单加氧酶进行多轮定向改造，获得不对称羟化活性提高的P450单加氧酶突变体N190D、N190F和N190F/V356L/A486E。其中，P450单加氧酶突变体N190F/V35