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Structural Insights into a Novel Esterase from the East Pacific Rise and Its Improved Thermostability by a Semirational Design
journal contribution
posted on 2021-01-15, 07:04 authored by Chunhua Zhu, Yayu Chen, Michail N. Isupov, Jennifer A. Littlechild, Lifang Sun, Xiaodong Liu, Qianchao Wang, Hui Gong, Panpan Dong, Na Zhang, Yunkun WuLipolytic 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 Å
resolution. The structure reveals a unique spatial arrangement and
orientation of the helix cap domain and α/β 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.
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Keywords
carboxyl ligandssequence alignment2.03 Å resolutionhelix cap domainsite pocketenzyme activityacyl chain substratesbinding affinityester bondsprotein engineeringEprEstK 114Penzyme engineeringthermostabilityp NP-butyrate esterEast Pacific RiseNovel Esterasehydrothermal ventesteraseStructural Insightsp NPX-ray structuresubstrate pocketshort-chain acyl groupssemirational designcrystal structuremarine-derived lipolytic enzymespesticide industriesfood processingSemirational Design Lipolytic enzymesMolecular docking analysis