反式环氧琥珀酸水解酶的热稳定性改造

鲍文娜; 姚锦锋; 潘海峰; 朱荣林; 厉欣颖; 廖鸿秀

doi:10.1631/jzus.B2500069

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Research Article | Updated：2026-01-27

反式环氧琥珀酸水解酶的热稳定性改造
Improving the thermal stability of trans-epoxysuccinate hydrolase
浙江大学学报（英文版）（B辑：生物医学和生物技术） 2026年27卷第1期页码：89-100
Affiliations：

1.School of Biological and Chemical Engineering, Zhejiang University of Science and Technology, Hangzhou 310023, China
2.School of Life Sciences and Health, Huzhou College, Huzhou 313000, China
Funds：

the Education Department of Zhejiang Province Scientific Research Project(Y202248484);the Huzhou Science and Technology Plan Project(2022GZ56)
DOI：10.1631/jzus.B2500069
中图分类号：
CSTR：
收稿：2025-02-09，

修回：2025-05-12，

网络出版：2025-12-22，

纸质出版：2026-01-27
Accepted：

鲍文娜,姚锦锋,潘海峰等.反式环氧琥珀酸水解酶的热稳定性改造[J].浙江大学学报（英文版）（B辑：生物医学和生物技术）,2026,27(01):89-100.

Wenna BAO, Jinfeng YAO, Haifeng PAN, et al. Improving the thermal stability of trans-epoxysuccinate hydrolase[J]. Journal of Zhejiang University-SCIENCE B, 2026, 27(1): 89-100.
鲍文娜,姚锦锋,潘海峰等.反式环氧琥珀酸水解酶的热稳定性改造[J].浙江大学学报（英文版）（B辑：生物医学和生物技术）,2026,27(01):89-100. DOI： 10.1631/jzus.B2500069.

Wenna BAO, Jinfeng YAO, Haifeng PAN, et al. Improving the thermal stability of trans-epoxysuccinate hydrolase[J]. Journal of Zhejiang University-SCIENCE B, 2026, 27(1): 89-100. DOI： 10.1631/jzus.B2500069.

摘要

本研究通过运用分子动力学模拟、B因子分析和饱和突变筛选等方法，提高了源自韩国假单胞菌的反式环氧琥珀酸水解酶（TESH）的热稳定性。在筛选出的11个影响该特性的突变体中，4个突变体活性有所提高，其中突变体A142C和S178Q的米氏常数（

）较低，其

cat

（催化常数）/

比值分别比野生型的高3.7倍和0.9倍。上述两个突变体在50 ℃下的半衰期（

https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=99131305&type=

3.47133350

https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=99131322&type=

4.14866638

）值相较野生型分别提高了107%和59%。分子对接和分子动力学模拟表明，与野生型相比，上述两个突变体与底物的相互作用更强，结合能更低，均方根偏差更小，且突变位点附近的静电势能更低，疏水性增强。综上，本研究结果可为内消旋酒石酸的工业化生产提供参考，具有现实意义。

Abstract

This study used molecular dynamics simulations

B-factor analysis

and saturation mutagenesis screening to enhance the thermal stability of the

trans

-epoxysuccinate hydrolase (TESH) derived from

Pseudomonas koreensis

. Eleven mutants that influence this characteristic were selected

yielding four mutants with improved activity. Among them

mutants A142C and S178Q exhibited lower Michaelis constant (

) values

and their

cat

ratios (

cat

catalytic constant) were 3.7 and 0.9 times higher than those of the wild type

respectively. The values of half-life at 50 ℃ (

https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=99131256&type=

3.47133350

https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=99131258&type=

4.14866638

) of the two mutants were increased by 107% and 59%

respectively

compared to the wild type. Molecular docking and molecular dynamics simulations indicated that the two mutants showed stronger substrate interaction

lower binding energy

and reduced root mean square deviation compared to the wild type

along with decreased electrostatic potential energy and increased hydrophobicity near their mutation sites. The study of protein thermal stability engineering and associated mechanisms provides a valuable reference and holds practical significance for the industrial production of

meso

-tartaric acid.

关键词

Keywords

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