Open AccessSpecific N‐demethylation of verapamil by cytochrome P450 from Streptomyces griseus ATCC 13273
Author(s) -
Shen Chen,
Liu Hanqing,
Dai Wenling,
Liu Xiufeng,
Liu Jihua,
Yu Boyang
Publication year - 2019
Publication title -
engineering in life sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.547
H-Index - 57
eISSN - 1618-2863
pISSN - 1618-0240
DOI - 10.1002/elsc.201800116
Subject(s) - demethylation , verapamil , streptomyces griseus , chemistry , cytochrome p450 , docking (animal) , microsome , biochemistry , cytochrome , stereochemistry , enzyme , streptomyces , biology , calcium , organic chemistry , bacteria , gene expression , genetics , dna methylation , gene , medicine , nursing
Norverapamil, the N‐demethylated derivative of verapamil, is a novel promising leading compound for attenuating multidrug resistance with less side effects compared with verapamil. However, the efficient synthetic method for norverapamil is absent. In this study, an innovative biotechnological method based on enzymatic catalysis was presented for the high‐efficient production of norverapamil. CYP105D1, a cytochrome P450 from Streptomyces griseus ATCC 13273, was identified to carry out a one‐step specific N‐demethylation of verapamil along with putidaredoxin reductase (Pdr) and putidaredoxin (Pdx) as the redox partner. Docking calculations rationalized the specific N‐demethylation observed in experiment and identified important amino acid residues for verapamil binding. Furthermore, a CYP105D1‐based whole‐cell system in E. coli BL21(DE3) was established and optimized for highly efficient N‐demethylation of verapamil. The bioconversion rate of verapamil by the whole cell system came up to 60.16% within 24 hours under the optimized conditions. These results demonstrated the high potential of CYP105D1‐based biocatalytic system for norverapamil production.