一种催化活性提高的12α-HSDH酶突变体及其用途

By performing site-directed mutagenesis on 12α-HSDH enzyme to improve its catalytic activity, the problems of insufficient enzyme activity and environmental pollution were solved, and efficient production of 12-ketochenodeoxycholic acid was achieved, which is suitable for industrial ursodeoxycholic acid production.

CN116445434BActive Publication Date: 2026-07-17ZHONGSHAN BAILING BIOTECHNOLOGY CO LTD

Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
ZHONGSHAN BAILING BIOTECHNOLOGY CO LTD
Filing Date
2022-12-21
Publication Date
2026-07-17

AI Technical Summary

Technical Problem

The current 12α-HSDH enzyme activity is insufficient, resulting in low production efficiency of ursodeoxycholic acid. Furthermore, the traditional chemical method causes serious pollution from oxidants, which does not meet the requirements of sustainable development.

Method used

By performing protein structure simulation, site-directed saturation mutagenesis, and Rosetta-assisted rational design on the 12α-HSDH enzyme derived from Clostridium sp. AF15-17LB, a 12α-HSDH mutant with enhanced catalytic activity was constructed by mutating serine at position 101 to alanine.

Benefits of technology

The mutant enzyme activity is increased by 1.5 times, and the substrate conversion rate and production rate are significantly improved, making it suitable for industrial production of 12-ketochenodeoxycholic acid, reducing production costs and environmental pollution.

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Abstract

本发明公开了一种催化活性提高的12α‑HSDH酶突变体及其用途,属于生物酶工程技术领域。本发明通过采用蛋白结构模拟技术、定点饱和突变以及Rosetta辅助对一种来源于梭菌属Clostridium sp.AF15‑17LB的辅酶NADP依赖型12α‑羟基类固醇脱氢酶进行定向突变改造,筛选得到活性显著提高的12α‑HSDH突变体,相较野生型单位酶活提高1.5倍,底物转化、生成率更好,对于提高12‑羰基鹅去氧胆酸的生产效率,对鹅去氧胆酸乃至熊去氧胆酸实现大量化生产具有重要意义。
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