6JSS

Structure of Geobacillus kaustophilus lactonase, Y99P mutant

6JSS の概要

• エントリーDOI: 10.2210/pdb6jss/pdb
• 分子名称: Phosphotriesterase, FE (III) ION, ZINC ION, ... (5 entities in total)
• 機能のキーワード: alpha-beta barrel, hydrolase
• 由来する生物種: Geobacillus kaustophilus (strain HTA426)
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 147808.51

構造登録者

Xue, B.,Yew, W.S. (登録日: 2019-04-08, 公開日: 2020-04-08, 最終更新日: 2023-11-22)

主引用文献

Go, M.K.,Zhao, L.N.,Xue, B.,Supekar, S.,Robinson, R.C.,Fan, H.,Yew, W.S.
Directed Computational Evolution of Quorum-Quenching Lactonases from the Amidohydrolase Superfamily.
Structure, 28:635-, 2020

PubMed Abstract: In this work, we present a generalizable directed computational evolution protocol to effectively reduce the sequence space to be explored in rational enzyme design. The protocol involves in silico mutation modeling and substrate docking to rapidly identify mutagenesis hotspots that may enhance an enzyme's substrate binding and overall catalysis. By applying this protocol to a quorum-quenching Geobacillus kaustophilus lactonase, GKL, we generated 1,881 single mutants and docked high-energy intermediates of nine acyl homoserine lactones onto them. We found that Phe28 and Tyr99 were two hotspots that produced most of the predicted top 20 mutants. Of the 180 enzyme-substrate combinations (top 20 mutants × 9 substrates), 51 (28%) exhibited enhanced substrate binding and 22 (12%) had better overall activity when compared with wild-type GKL. X-ray crystallographic studies of Y99C and Y99P provided rationalized explanations for the enhancement in enzyme function and corroborated the utility of the protocol.

PubMed: 32320671
DOI: 10.1016/j.str.2020.03.011

実験手法

X-RAY DIFFRACTION (2.16 Å)