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Characterization of a Self‐Sufficient Cytochrome P450 Monooxygenase from Deinococcus apachensis for Enantioselective Benzylic Hydroxylation
Author(s) -
Xie Lingzhi,
Chen Ke,
Cui Haibo,
Wan Nanwei,
Cui Baodong,
Han Wenyong,
Chen Yongzheng
Publication year - 2020
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201900691
Subject(s) - hydroxylation , enantioselective synthesis , monooxygenase , stereochemistry , chemistry , escherichia coli , cytochrome p450 , subfamily , recombinant dna , substrate (aquarium) , cofactor , biochemistry , enzyme , biology , catalysis , gene , ecology
Abstract A self‐sufficient cytochrome P450 monooxygenase from Deinococcus apachensis (P450DA) was identified and successfully overexpressed in Escherichia coli BL21(DE3). P450DA would be a member of the CYP102D subfamily and assigned as CYP102D2 according to the phylogenetic tree and sequence alignment. Purification and characterization of the recombinant P450DA indicated both NADH and NADPH could be used by P450DA as a reducing cofactor. The recombinant E. coli (P450DA) strain was functionally active, showing excellent enantioselectivity for benzylic hydroxylation of methyl 2‐phenylacetate. Further substrate scope studies revealed that P450DA is able to catalyze benzylic hydroxylation of a variety of compounds, affording the corresponding chiral benzylic alcohols in 86–99 % ee and 130–1020 total turnover numbers.