6SYA

STRUCTURE OF S192A-ESTER-HYDROLASE EH3 FROM THE METAGENOME OF MARINE SEDIMENTS AT MILAZZO HARBOR (SICILY, ITALY) COMPLEXED WITH METHYL (2R)-2-PHENYLPROPANOATE

6SYA の概要

• エントリーDOI: 10.2210/pdb6sya/pdb
• 分子名称: Esterase, GLYCEROL, methyl (2~{R})-2-phenylpropanoate, ... (4 entities in total)
• 機能のキーワード: ester hydrolase, complex, hydrolase
• 由来する生物種: uncultured bacterium
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 81468.22

構造登録者

Cea-Rama, I.,Sanz-Aparicio, J. (登録日: 2019-09-27, 公開日: 2021-04-07, 最終更新日: 2024-01-24)

主引用文献

Cea-Rama, I.,Coscolin, C.,Katsonis, P.,Bargiela, R.,Golyshin, P.N.,Lichtarge, O.,Ferrer, M.,Sanz-Aparicio, J.
Structure and evolutionary trace-assisted screening of a residue swapping the substrate ambiguity and chiral specificity in an esterase.
Comput Struct Biotechnol J, 19:2307-2317, 2021

PubMed Abstract: Our understanding of enzymes with high substrate ambiguity remains limited because their large active sites allow substrate docking freedom to an extent that seems incompatible with stereospecificity. One possibility is that some of these enzymes evolved a set of evolutionarily fitted sequence positions that stringently allow switching substrate ambiguity and chiral specificity. To explore this hypothesis, we targeted for mutation a serine ester hydrolase (EH) that exhibits an impressive 71-substrate repertoire but is not stereospecific ( 50%). We used structural actions and the computational evolutionary trace method to explore specificity-swapping sequence positions and hypothesized that position I244 was critical. Driven by evolutionary action analysis, this position was substituted to leucine, which together with isoleucine appears to be the amino acid most commonly present in the closest homologous sequences (max. identity, . 67.1%), and to phenylalanine, which appears in distant homologues. While the I244L mutation did not have any functional consequences, the I244F mutation allowed the esterase to maintain a remarkable 53-substrate range while gaining stereospecificity properties ( 99.99%). These data support the possibility that some enzymes evolve sequence positions that control the substrate scope and stereospecificity. Such residues, which can be evolutionarily screened, may serve as starting points for further designing substrate-ambiguous, yet chiral-specific, enzymes that are greatly appreciated in biotechnology and synthetic chemistry.

PubMed: 33995922
DOI: 10.1016/j.csbj.2021.04.041

実験手法

X-RAY DIFFRACTION (2.27 Å)