biostudies-literatureCheng Cthe Jiangsu Provincial Association for Science and Technology Youth Talent Support ProjecNational Natural Science Foundation Youth FundNatural Science Foundation of Jiangsu Province3https://www.ebi.ac.uk/biostudies/studies/S-EPMC12779709biostudies-literatureUnknown110(1)The overexpression of proteins in Escherichia coli often results in the formation of inclusion bodies, which are biologically inactive, especially for proteins with exposed hydrophobic surfaces. Solubilization of inclusion bodies (IBs) and subsequent refolding is essential for obtaining correctly folded and active protein. However, protein refolding involves multiple steps-namely isolation, solubilization, and refolding-which is a labor-intensive process. In this study, we developed a strategy for soluble production and protein refolding. A fusion tag was applied to Burkholderia ambifaria lipase YCJ01, enabling abundant soluble expression in E. coli. Despite this, the soluble protein exhibited only partial enzymatic activity, suggesting an unfolded state of soluble lipase YCJ01. Lipase actApplied microbiology and biotechnologyCosolvent-induced spontaneous refolding of lipase.PMC12779709TJ-2023-02122308159BK20220335Cheng CQiu YCui LJiang TSu YWang JHe BfalseCosolvent-induced spontaneous refolding of lipase.The overexpression of proteins in Escherichia coli often results in the formation of inclusion bodies, which are biologically inactive, especially for proteins with exposed hydrophobic surfaces. Solubilization of inclusion bodies (IBs) and subsequent refolding is essential for obtaining correctly folded and active protein. However, protein refolding involves multiple steps-namely isolation, solubilization, and refolding-which is a labor-intensive process. In this study, we developed a strategy for soluble production and protein refolding. A fusion tag was applied to Burkholderia ambifaria lipase YCJ01, enabling abundant soluble expression in E. coli. Despite this, the soluble protein exhibited only partial enzymatic activity, suggesting an unfolded state of soluble lipase YCJ01. Lipase act2026-01-01T00:00:00Z2026 Jan2026-07-15T07:44:57.813Z2026-07-01T03:07:59.393ZS-EPMC127797094149526910.1007/s00253-025-13660-6