5HC2

Structure of esterase Est22 mutant-S188A with p-nitrophenol

Summary for 5HC2

• Entry DOI: 10.2210/pdb5hc2/pdb
• Related: 5HC0 5HC3 5HC4 5HC5
• Descriptor: Lipolytic enzyme, P-NITROPHENOL, IMIDAZOLE, ... (4 entities in total)
• Functional Keywords: esterase, est22, hydrolase
• Biological source: uncultured bacterium
• Total number of polymer chains: 4
• Total formula weight: 157168.60

Authors

Li, J.,Huang, J. (deposition date: 2016-01-04, release date: 2017-01-18, Last modification date: 2024-03-20)

Primary citation

Huang, J.,Huo, Y.Y.,Ji, R.,Kuang, S.,Ji, C.,Xu, X.W.,Li, J.
Structural insights of a hormone sensitive lipase homologue Est22.
Sci Rep, 6:28550-28550, 2016

PubMed Abstract: Hormone sensitive lipase (HSL) catalyzes the hydrolysis of triacylglycerols into fatty acids and glycerol, thus playing key roles in energy homeostasis. However, the application of HSL serving as a pharmaceutical target and an industrial biocatalyst is largely hampered due to the lack of high-resolution structural information. Here we report biochemical properties and crystal structures of a novel HSL homologue esterase Est22 from a deep-sea metagenomic library. Est22 prefers short acyl chain esters and has a very high activity with substrate p-nitrophenyl butyrate. The crystal structures of wild type and mutated Est22 with its product p-nitrophenol are solved with resolutions ranging from 1.4 Å to 2.43 Å. The Est22 exhibits a α/β-hydrolase fold consisting with a catalytic domain and a substrate-recognizing cap domain. Residues Ser188, Asp287, and His317 comprise the catalytic triad in the catalytic domain. The p-nitrophenol molecule occupies the substrate binding pocket and forms hydrogen bonds with adjacent residues Gly108, Gly109, and Gly189. Est22 exhibits a dimeric form in solution, whereas mutants D287A and H317A change to polymeric form, which totally abolished its enzymatic activities. Our study provides insights into the catalytic mechanism of HSL family esterase and facilitates the understanding for further industrial and biotechnological applications of esterases.

PubMed: 27328716
DOI: 10.1038/srep28550

Experimental method

X-RAY DIFFRACTION (1.986 Å)