Department of Bioengineering Shanghai Institute of TechnologyFengxian, Shanghai, 201418, China.
Acta Pharm. 2021 Mar 1;71(1):99-114. doi: 10.2478/acph-2021-0004.
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 17-18 % 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 poten tial for application in the industrial production of eplerenone.
依普利酮是一种保护心血管系统的药物。11α-羟基坎利酮是依普利酮合成的关键中间体。我们发现，尽管曲霉 MF018 中的细胞色素 P450（CYP）酶系统可以催化坎利酮转化为 11α-羟基坎利酮，但生物催化效率较低。为了提高 11α-羟基坎利酮的生产效率，根据全基因组测序结果，预测并克隆了 MF018 的 CYP 单加氧酶编码基因。通过同源重组，获得了 CYP 单加氧酶高表达的重组曲霉 MF010 菌株。该重组菌在 60 小时内无需添加有机溶剂或表面活性剂即可达到 93%的生物催化率，比 MF018 菌株高 17-18%。此外，与 MF018 菌株相比，MF010 菌株的生物催化时间缩短了 30 小时以上。这些结果表明，重组曲霉 MF010 可以克服底物生物催化效率的限制，因此具有在依普利酮工业生产中应用的巨大潜力。
