Heterologous expression and substrate specificity of ketoreductase domain in bacillaene polyketide synthase.
- Author:
Xiaohui SUN
;
Chengchuan CHE
;
Junjie JI
;
Jianting ZHENG
;
Ge YANG
- Publication Type:Journal Article
- MeSH:
Bacterial Proteins;
genetics;
metabolism;
Catalysis;
Cyclohexanones;
metabolism;
Escherichia coli;
enzymology;
Polyketide Synthases;
genetics;
metabolism;
Protein Structure, Tertiary;
Substrate Specificity;
omega-Chloroacetophenone;
metabolism
- From:
Chinese Journal of Biotechnology
2015;31(9):1355-1362
- CountryChina
- Language:Chinese
-
Abstract:
The ketoreductase (KR) domain in the first extending module of the polyketide synthase (PKS) catalyzes the reductions of both an α-keto group and a β-keto group in the biosynthesis of bacillaene, suggesting the intrinsic substrate promiscuity. In order to further investigate the substrate specificity, the KR domain (BacKR1) was heterologously overexpressed in Escherichia coli. In vitro enzymatic analysis showed that only one of the four diastereomers was formed in the reduction of the racemic (±)-2-methyl-3-oxopentanoyl-N-acetylcysteamine thioester catalyzed by BacKR1. In addition, BacKR1 was revealed to catalyze the reductions of cyclohexanone and p-chloroacetophenone, indicating the potential of KR domians of PKSs as biocatalysts.
