Regio- and stereoselective hydroxylation of testosterone by cytochrome P450 from Streptomyces griseus ATCC 13273. Issue 2 (4th March 2021)

• Record Type: Journal Article
• Authors: Zhu, Jinqian; Shen, Chen; Zhao, Wanli; Liu, Xiufeng; Liu, Jihua; Yu, Boyang
• Is Part Of: Biocatalysis and biotransformation. Volume 39:Issue 2(2021)
• Page Start: 130
• Page End: 137
• Publication Date: 2021-03-04
• Subjects: Cytochrome P450 -- testosterone -- biotransformation -- hydroxylation -- regio- and stereoselectivity; Enzymes -- Biotechnology -- Periodicals; Enzymes -- Industrial applications -- Periodicals; Biotransformation (Metabolism) -- Periodicals; 660.63
• DOI: 10.1080/10242422.2020.1799990
• Languages: English
• ISSNs: 1024-2422

Abstract

Abstract: 16α-Hydroxylated steroids are important precursors in the synthesis of high value steroidal drugs such as glucocorticoids in the pharmaceutical industry. In this study, an innovative biotechnological method based on enzymatic catalysis was presented for the high-efficient production of 16α-hydroxyl-testosterone. CYP154C, a cytochrome P450 from Streptomyces griseus ATCC 13273, was identified to carry out a one-step regio- and stereoselective hydroxylation of testosterone along with putidaredoxin reductase (PDR) and putidaredoxin (Pdx) as the redox partner. The K m and k cat values of CYP154C for testosterone were 23.40 ± 3.69 μM and 4.75 min −1, respectively. The catalytic efficiency ( k cat / K m ) for testosterone was 12.18 s −1 ·μM −1 . Using a CYP154-based whole-cell system in E. coli BL21(DE3), high efficiency of hydroxylation of testosterone for production of 16α-hydroxyl-testosterone was achieved. The bioconversion rate of testosterone (50 mg/L) by the whole cell system came up to 75 ± 2.87% within 24 h. These results demonstrated the high potential of CYP154-based biocatalytic system for 16α-hydroxyl-testosterone production.