安徽医科大学学报2025，Vol.60Issue(8)：1365-1372,8.DOI:10.19405/j.cnki.issn1000-1492.2025.08.001
羰基还原酶ChKRED20可溶性表达的提升及酶学性质研究
Enhancing solubility expression of carbonyl reductase ChKRED20 and characterization of its enzymatic properties
摘要
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.关键词
羰基还原酶/ChKRED20/His-tag/酶学性质/大肠埃希菌/蛋白可溶性Key words
carbonyl reductase/ChKRED20/His-tag/enzymatic properties/Escherichia coli/protein solubility分类
医药卫生引用本文复制引用
沈悦,郑红,范新炯..羰基还原酶ChKRED20可溶性表达的提升及酶学性质研究[J].安徽医科大学学报,2025,60(8):1365-1372,8.基金项目
国家自然科学基金项目(编号:82173725、31400680) （编号:82173725、31400680）
高校中青年教师培养行动项目(编号:DTR2024005) National Natural Science Foundation of China(Nos.82173725,31400680) （编号:DTR2024005）
Young and Middle-aged Teacher Develop Program in Colleges and Universities(No.DTR2024005) （No.DTR2024005）
