Engineering 7β-Hydroxysteroid Dehydrogenase for Enhanced Ursodeoxycholic Acid Production by Multiobjective Directed Evolution

2017/01/24 by Ming‐Min Zheng, Ming-Min Zheng, Ke-Cai Chen +7 · 9 citations
Biochemistry, Genetics and Molecular Biology · Pharmacology, Toxicology and Pharmaceutics · #Biochemical and Molecular Research #Enzyme Catalysis and Immobilization #Pharmacogenetics and Drug Metabolism

paper · doi:10.1021/acs.jafc.6b05428

Abstract

Ursodeoxycholic acid (UDCA) is the main active ingredient of natural bear bile powder with multiple pharmacological functions. 7β-Hydroxysteroid dehydrogenase (HSDH) is a key biocatalyst for the synthesis of UDCA. However, all the 7β-HSDHs reported commonly suffer from poor activity and thermostability, resulting in limited productivity of UDCA. In this study, a multiobjective directed evolution (MODE) strategy was proposed and applied to improve the activity, thermostability, and pH optimum of a 7β-HSDH. The best variant (V 3–1 ) showed a specific activity 5.5-fold higher than and a half-life 3-fold longer than those of the wild type. In addition, the pH optimum of the variant was shifted to a weakly alkaline value. In the cascade reaction, the productivity of UDCA with V 3–1 increased to 942 g L –1 day –1, in contrast to 141 g L –1 day –1 with the wild type. Therefore, this study provides a useful strategy for improving the catalytic efficiency of a key enzyme that significantly facilitated the bioproduction of UDCA.

Cited by

• Latest development in the synthesis of ursodeoxycholic acid (UDCA): a critical review. [europepmc]
• Functional tunability from a distance: Rheostat positions influence allosteric coupling between two distant binding sites. [europepmc]
• Biological synthesis of ursodeoxycholic acid. [europepmc]
• A gut microbiota-bile acid axis promotes intestinal homeostasis upon aspirin-mediated damage. [europepmc]
• A green strategy to produce potential substitute resource for bear bile using engineered Saccharomyces cerevisiae. [europepmc]
• Engineering of a hydroxysteroid dehydrogenase with simultaneous enhancement in activity and thermostability for efficient biosynthesis of ursodeoxycholic acid. [europepmc]
• Cost-effective whole-cell biosynthesis of ursodeoxycholic acid using engineered Escherichia coli with a multienzyme cascade. [europepmc]
• Faecal metabolites as a readout of habitual diet capture dietary interactions with the gut microbiome. [europepmc]
• Advances in Pharmacological Activities, Biosynthesis, and Structural Modification of Ursodeoxycholic Acid (UDCA): A Review. [europepmc]

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