Overexpression and truncation of a novel cold-adapted lipase with improved enzymatic characteristics

Yue Zhang; Yu Gao; Jiahui Chen; Fang Yu; Yongming Bao

doi:10.1016/j.pep.2023.106376

Overexpression and truncation of a novel cold-adapted lipase with improved enzymatic characteristics

Protein Expr Purif. 2024 Feb:214:106376. doi: 10.1016/j.pep.2023.106376. Epub 2023 Oct 14.

Authors

Yue Zhang¹, Yu Gao², Jiahui Chen², Fang Yu², Yongming Bao³

Affiliations

¹ School of Biological Engineering, Dalian Polytechnic University, Dalian, 116034, China. Electronic address: zhangyue124@126.com.
² School of Biological Engineering, Dalian Polytechnic University, Dalian, 116034, China.
³ School of Bioengineering, Dalian University of Technology, Dalian, 116024, China. Electronic address: biosci@dlut.edu.cn.

PMID: 37839629
DOI: 10.1016/j.pep.2023.106376

Abstract

The novel cold-adapted lipase (Lip ZC12) derived from Psychrobacter sp. ZY124 exhibited higher catalytic activity at 20-40 °C, the whole gene was then sequenced, analyzed, and overexpressed. However, its intrinsic structural characteristics lead to a decreased affinity toward the substrate, thus limiting the improvement of catalytic efficiency. Modeling the homologous structure and simulating the binding process of Lip ZC12 with the substrate. It was found that truncated lid (lip-Δlid) could not only increase the k_cat, but also significantly enhance the substrate affinity, the substrate affinity and catalytic efficiency of Lip ZC12 modified by lid truncation were significantly improved. The results revealed that the k_cat/K_m value of lip-Δlid was 1.6 times higher than that of free lipase. This improved catalytic performance of cold-adapted lipase, and these findings laid an important foundation for further application.

Keywords: Clone and expression; Cold-adapted lipase; Substrate affinity; Truncation.

MeSH terms

Catalysis
Cold Temperature*
Lipase* / metabolism
Substrate Specificity

Substances

Lipase