Acta Pharm. 71 (2021) 99-114

 

full paper

Original research paper

 

Improved 11α-hydroxycanrenone production by modification of cytochrome P450 monooxygenase gene in Aspergillus ochraceus

QIANQIAN LI, LI SHI, YINGYING LIU, SHIMIN GUAN, SHUO ZHANG,
BAOGUO CAI and SHAOFENG RONG

rongshaofeng@163.com

Department of Bioengineering, Shanghai Institute of Technology, Fengxian, Shanghai, 201418, China

Accepted March 13, 2020

Published online April 10, 2020

 

Eplerenone is a drug that protects the cardiovascular system. 11α-Hydroxycanrenone is a key intermediate in eplerenone synthesis. We found that although the cytochrome P450 (CYP) enzyme system in Aspergillus ochraceus strain MF018 could catalyse the conversion of canrenone to 11α-hydroxycanrenone, its biocatalytic efficiency is low. To improve the efficiency of 11α-hydroxycanrenone production, the CYP monooxygenase-coding gene of MF018 was predicted and cloned based on whole-genome sequencing results. A recombinant A. ochraceus strain MF010 with the high expression of CYP monooxygenase was then obtained through homologous recombination. The biocatalytic rate of this recombinant strain reached 93 % at 60 h without the addition of organic solvents or surfactants and was 1718 % higher than that of the MF018 strain. Moreover, the biocatalytic time of the MF010 strain was reduced by more than 30 h compared with that of the MF018 strain. These results show that the recombinant A. ochraceus strain MF010 can overcome the limitation of substrate biocatalytic efficiency and thus holds a high potential for application in the industrial production of eplerenone.

 

Keywords: Aspergillus ochraceus, P450 monooxygenase, homologous recombination, canrenone, 11α-hydroxycanrenone