6Z69

A novel metagenomic alpha/beta-fold esterase

6Z69 の概要

• エントリーDOI: 10.2210/pdb6z69/pdb
• 分子名称: Acetyl esterase/lipase, N-HEXYLPHOSPHONATE ETHYL ESTER, 7-hydroxy-4-methyl-2H-chromen-2-one, ... (5 entities in total)
• 機能のキーワード: alpha/beta-fold hydrolase; esterase; metagenome; inhibitor-bound; substrate promiscuity, hydrolase
• 由来する生物種: Pseudonocardia thermophila
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 81986.53

構造登録者

Bollinger, A.,Thies, S.,Hoeppner, A.,Kobus, S.,Jaeger, K.-E.,Smits, S.H.J. (登録日: 2020-05-28, 公開日: 2020-12-30, 最終更新日: 2024-01-24)

主引用文献

Hoppner, A.,Bollinger, A.,Kobus, S.,Thies, S.,Coscolin, C.,Ferrer, M.,Jaeger, K.E.,Smits, S.H.J.
Crystal structures of a novel family IV esterase in free and substrate-bound form.
Febs J., 288:3570-3584, 2021

PubMed Abstract: Bacterial lipolytic enzymes of family IV are homologs of the mammalian hormone-sensitive lipases (HSL) and have been successfully used for various biotechnological applications. The broad substrate specificity and ability for enantio-, regio-, and stereoselective hydrolysis are remarkable features of enzymes from this class. Many crystal structures are available for esterases and lipases, but structures of enzyme-substrate or enzyme-inhibitor complexes are less frequent although important to understand the molecular basis of enzyme-substrate interaction and to rationalize biochemical enzyme characteristics. Here, we report on the structures of a novel family IV esterase isolated from a metagenomic screen, which shows a broad substrate specificity. We solved the crystal structures in the apo form and with a bound substrate analogue at 1.35 and 1.81 Å resolution, respectively. This enzyme named PtEst1 hydrolyzed more than 60 out 96 structurally different ester substrates thus being substrate promiscuous. Its broad substrate specificity is in accord with a large active site cavity, which is covered by an α-helical cap domain. The substrate analogue methyl 4-methylumbelliferyl hexylphosphonate was rapidly hydrolyzed by the enzyme leading to a complete inactivation caused by covalent binding of phosphinic acid to the catalytic serine. Interestingly, the alcohol leaving group 4-methylumbelliferone was found remaining in the active site cavity, and additionally, a complete inhibitor molecule was found at the cap domain next to the entrance of the substrate tunnel. This unique situation allowed gaining valuable insights into the role of the cap domain for enzyme-substrate interaction of esterases belonging to family IV. DATABASE: Structural data of PtEst1 are available in the worldwide protein data bank (https://www.rcsb.org) under the accession codes: 6Z68 (apo-PtEst1) and 6Z69 (PtEst1-inhibitor complex).

PubMed: 33342083
DOI: 10.1111/febs.15680

実験手法

X-RAY DIFFRACTION (1.81 Å)