Multi-Enzymatic One-Pot Reduction of Lithocholic Acid to Ursodeoxycholic Acid
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.
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