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A Customizable Approach for the Enzymatic Production and Purification of Diterpenoid Natural Products

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

Ming-Min Zheng¹, Ke-Cai Chen¹, Ru-Feng Wang¹

¹State Key Laboratory of Bioreactor Engineering and ‡Shanghai Collaborative Innovation Center for Biomanufacturing Technology, East China University of Science and Technology , Shanghai 200237, P. R. China.

Journal of Agricultural and Food Chemistry

|January 25, 2017

Summary

Researchers enhanced a key enzyme, 7β-Hydroxysteroid dehydrogenase (HSDH), using multiobjective directed evolution. This significantly boosted the bioproduction of ursodeoxycholic acid (UDCA), a valuable pharmaceutical ingredient.

Keywords:

7β-hydroxysteroid dehydrogenase biocatalysis cascade reaction multiobjective directed evolution ursodeoxycholic acid

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Area of Science:

Biotechnology
Enzyme Engineering
Biocatalysis

Background:

Ursodeoxycholic acid (UDCA) is a key pharmaceutical compound derived from bile.
7β-Hydroxysteroid dehydrogenase (HSDH) is crucial for UDCA synthesis but exhibits limitations in activity and stability.
Existing HSDH enzymes hinder efficient and scalable UDCA production.

Purpose of the Study:

To improve the activity, thermostability, and pH profile of 7β-HSDH.
To enhance the overall efficiency of UDCA bioproduction.
To develop a robust enzyme for industrial applications.

Main Methods:

Multiobjective directed evolution (MODE) was employed to engineer 7β-HSDH.
The enzyme variant V3-1 was selected based on improved catalytic properties.
Performance was evaluated in a cascade reaction for UDCA synthesis.

Main Results:

The best variant (V3-1) exhibited a 5.5-fold increase in specific activity and a 3-fold longer half-life compared to the wild type.
The variant's optimal pH shifted to a weakly alkaline range.
UDCA production reached 942 g L⁻¹ day⁻¹ with V3-1, a substantial improvement over the wild type's 141 g L⁻¹ day⁻¹.

Conclusions:

The MODE strategy successfully enhanced 7β-HSDH performance.
The engineered enzyme significantly boosts UDCA bioproduction efficiency.
This work offers a valuable approach for enzyme improvement in biocatalysis.