Adaptation of the electron transport chain improves the biocatalytic efficiency of progesterone 17α hydroxylation.
- Author:
Lanlan WANG
1
;
Xin ZHAO
1
;
Jie LI
1
;
Jiaying AI
1
;
Jing SUN
1
;
Shuhong MAO
1
Author Information
- Publication Type:Journal Article
- Keywords: 17α hydroxylase; electronic transmission chain; progesterone; protein mutation
- MeSH: Animals; Cattle; Progesterone/metabolism*; Hydroxylation; Biocatalysis; Electron Transport; Mixed Function Oxygenases/metabolism*
- From: Chinese Journal of Biotechnology 2023;39(11):4608-4620
- CountryChina
- Language:Chinese
- Abstract: 17α hydroxylase is a key enzyme for the conversion of progesterone to prepare various progestational drug intermediates. To improve the specific hydroxylation capability of this enzyme in steroid biocatalysis, the CYP260A1 derived from cellulose-mucilaginous bacteria Sorangium cellulosum Soce56 and the Fpr and bovine adrenal-derived Adx4-108 derived from Escherichia coli str. K-12 were used to construct a new electron transfer system for the conversion of progesterone. Selective mutation of CYP260A1 resulted in a mutant S276I with significantly enhanced 17α hydroxylase activity, and the yield of 17α-OH progesterone reached 58% after optimization of the catalytic system in vitro. In addition, the effect of phosphorylation of the ferredoxin Adx4-108 on 17α hydroxyl activity was evaluated using a targeted mutation technique, and the results showed that the mutation Adx4-108T69E transferred electrons to S276I more efficiently, which further enhanced the catalytic specificity in the C17 position of progesterone, and the yield of 17α-OH progesterone was eventually increased to 74%. This study provides a new option for the production of 17α-OH progesterone by specific transformation of bacterial-derived 17α hydroxylase, and lays a theoretical foundation for the industrial production of progesterone analogs using biotransformation method.
