Journal
CHEMBIOCHEM
Volume 21, Issue 13, Pages 1820-1825Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.201900691
Keywords
characterization; chiral benzylic alcohols; cytochrome P450 monooxygenase; enantioselectivity; hydroxylation
Funding
- National Natural Science Foundation of China [21961048, 21562054]
- Guizhou Science and Technology Department [QKHRC-2016-4029, QKHPTRC-2016-5801, QKHJC-2019-1467, QKHZC-2019-2579]
- Program for Outstanding Youth of Zunyi Medical University [17zy-001]
- National First-Rate Construction Discipline of Guizhou Province (Pharmacy) [YLXKJS-YX-04]
- Fifth Batch of Talent Base in Guizhou Province (2016)
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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.
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