Enhancing solubility expression of carbonyl reductase ChKRED20 and characterization of its enzymatic properties
10.19405/j.cnki.issn1000-1492.2025.08.001
- VernacularTitle:羰基还原酶 ChKRED20 可溶性表达的提升及酶学性质研究
- Author:
Yue Shen
1
;
Hong Zheng
1
;
Xinjiong Fan
1
Author Information
1. Stem Cell and Tissue Research Center , School of Basic Medicine , Anhui Medical University , Hefei 230032
- Publication Type:Journal Article
- Keywords:
carbonyl reductase;
ChKRED20;
His⁃tag;
enzymatic properties;
Escherichia coli;
protein solubility
- From:
Acta Universitatis Medicinalis Anhui
2025;60(8):1365-1372
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To enhance the solubility expression of carbonyl reductase ChKRED20 and investigate its enzymatic properties.
Methods :The ChKRED20 gene was synthesized and transformed into Escherichia coli BL21(DE3) for heterologous expression. The solubility of the protein was evaluated by adding or removing His⁃tag ,changing its position , and adjusting the induction conditions. Further optimization was performed by varying the in⁃duction time , temperature and IPTG concentration. Subsequently , the enzymatic properties of the recombinant en⁃zyme were examined.
Results:The ChKRED20 was cloned within the Escherichia coli system to achieve soluble protein expression . This was achieved by removing the His_tag and optimizing expression conditions . Compared to enzymes with the N_terminal and C _terminal His_tags , the enzyme activity of ChKRED20 without His_tag increased by 0. 77 and 1 . 28 times , respectively. The optimal temperature and pH for enzyme activity were 55 ℃ and 8. 0 , respectively. Furthermore , the enzyme demonstrated good adaptability in higher temperature and alkaline environ_ ments .
Conclusion:The removal of the His_tag and the optimization of expression conditions can significantly im_ prove the soluble expression of ChKRED20 in Escherichia coli .
- Full text:2026040321520433178羰基还原酶ChKRED20可溶性表达的提升及酶学性质研究_沈悦.pdf
