Different strategies to enhance the solubility of acylsucrose transferase PaxASAT1 expression in Escherichia coli
Year: 2026
Type: article
Abstract: Heterologous expression of soluble proteins is a crucial method for studying gene function and enzyme structure. Escherichia coli is a widely used host for expressing foreign genes from plants, animals, and microorganisms. PaxASAT1, a member of the BAHD acyltransferase family, plays a pivotal role in the initial step of acylsucrose biosynthesis by catalyzing the formation of mono-acylsucrose from sucrose and fatty acyl-CoA. In this study, PaxASAT1 was expressed in BL21(DE3) but with low solubility. Several strategies were evaluated to enhance soluble expression of PaxASAT1, including codon optimization, chaperone co-expression, soluble-tag fusion, and SEP-tag utilization. Results demonstrated that the inclusion of molecular chaperones like DnaK resulted in a slight improvement in the solubility of PaxASAT1. SUMO and MBP tag fusions significantly enhanced PaxASAT1 soluble expression. Notably, C9K-modified SEP tags substantially increased the solubility of PaxASAT1. This study provides a theoretical foundation for the soluble expression of acylsucrose acyltransferase in E. coli, paving the way for further research and applications in the field.
Authors: Li Han, Xueming Yu, Yibo Ning, Chenhui Wang, Changtong Lu, Ziqiang Dong, Chang Liu, Xiongjun Ni, Zhifei Chen
Institutions: Zhengzhou University of Light Industry, China Tobacco
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $2,700
OpenAlex ID: W7160821747