Abstract:Ursodeoxycholic acid is an important therapeutic bile acid, especially in the treatment of cholecystitis and the prevention of cholesterol-type gallstones. The authors produced ursodeoxycholic acid through regioselective and stereoselective hydroxylation at the 7β-position of lithocholic acid using the P450 monooxygenase CYP107D1 (OleP) in E. coli whole-cell system. Ursodeoxycholic acid was biosynthesized using an efficient one-step method, avoiding the cruel practice of extracting bile from live bear and the inefficiencies of chemical synthesis. Through the design of the co-expression combination of the mutant F84Q/S240A/V291G (OlePs) of OleP and the redox chaperone, a recombinant expression strain Escherichia coli BL21/PDR-PDX-SD-AS-OlePs was developed. Utilizing this recombinant strain, the whole-cell catalytic reaction of E. coli BL21/PDR-PDX-SD-AS-OlePs led to a substantial increase in a significantly increased yield of ursodeoxycholic acid yield at 88.28%, which was 73.96% higher than that of without the co-expression of redox chaperone. The author has made progress in the preparation of ursodeoxycholic acid through one-step reduction of lithocholic acid, offering possibilities for the synthesis of ursodeoxycholic acid from biowaste lithocholic acid.
