Structure Report for: 7C4D

Chunhua Zhu, Yayu Chen, Michail N. Isupov, Jennifer A. Littlechild, Lifang Sun, Xiaodong Liu, Qianchao Wang, Hui Gong, Panpan Dong, Na Zhang, and Yunkun Wu

Title: Structural Insights into a Novel Esterase from the East Pacific Rise and Its Improved Thermostability by a Semirational Design
Zhu C, Chen Y, Isupov MN, Littlechild JA, Sun L, Liu X, Wang Q, Gong H, Dong P and Wu Y <1 more author(s)> Zhu C, Chen Y, Isupov MN, Littlechild JA, Sun L, Liu X, Wang Q, Gong H, Dong P, Zhang N, Wu Y (- 1)
Ref: Journal of Agricultural and Food Chemistry, :, 2021 : PubMed
Abstract


ESTHER: Zhu_2021_J.Agric.Food.Chem__
PubMedSearch: Zhu 2021 J.Agric.Food.Chem
PubMedID: 33445864
Gene_locus related to this paper: 9bact-a0a2s1gux0

Abstract

Lipolytic enzymes are essential biocatalysts in food processing as well as pharmaceutical and pesticide industries, catalyzing the cleavage of ester bonds in a variety of acyl chain substrates. Here, we report the crystal structure of an esterase from the deep-sea hydrothermal vent of the East Pacific Rise (EprEst). The X-ray structure of EprEst in complex with the ligand, acetate, has been determined at 2.03 resolution. The structure reveals a unique spatial arrangement and orientation of the helix cap domain and alpha/beta hydrolase domain, which form a substrate pocket with preference for short-chain acyl groups. Molecular docking analysis further demonstrated that the active site pocket could accommodate p-nitrophenyl (pNP) carboxyl ligands of varying lengths (>=6 C atoms), with pNP-butyrate ester predicted to have the highest binding affinity. Additionally, the semirational design was conducted to improve the thermostability of EprEst by enzyme engineering based on the established structure and multiple sequence alignment. A mutation, K114P, introduced in the hinge region of the esterase, which displayed increased thermostability and enzyme activity. Collectively, the structural and functional data obtained herein could be used as basis for further protein engineering to ultimately expand the scope of industrial applications of marine-derived lipolytic enzymes.