4XBY

Crystal Structure of the L74F/M78V/I80V/L114F mutant of LEH complexed with cyclopentene oxide

4XBY の概要

• エントリーDOI: 10.2210/pdb4xby/pdb
• 関連するPDBエントリー: 4xbt 4xbx
• 分子名称: Limonene-1,2-epoxide hydrolase, (1R,5S)-6-oxabicyclo[3.1.0]hexane (3 entities in total)
• 機能のキーワード: epoxide hydrolase, hydrolase
• 由来する生物種: Rhodococcus erythropolis
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 139291.51

構造登録者

Kong, X.D.,Sun, Z.,Xu, J.H.,Reetz, M.T.,Zhou, J. (登録日: 2014-12-17, 公開日: 2015-07-15, 最終更新日: 2023-11-08)

主引用文献

Sun, Z.,Lonsdale, R.,Kong, X.D.,Xu, J.H.,Zhou, J.,Reetz, M.T.
Reshaping an Enzyme Binding Pocket for Enhanced and Inverted Stereoselectivity: Use of Smallest Amino Acid Alphabets in Directed Evolution
Angew.Chem.Int.Ed.Engl., 54:12410-12415, 2015

PubMed Abstract: Directed evolution based on saturation mutagenesis at sites lining the binding pocket is a commonly practiced strategy for enhancing or inverting the stereoselectivity of enzymes for use in organic chemistry or biotechnology. However, as the number of residues in a randomization site increases to five or more, the screening effort for 95 % library coverage increases astronomically until it is no longer feasible. We propose the use of a single amino acid for saturation mutagenesis at superlarge randomization sites comprising 10 or more residues. When used to reshape the binding pocket of limonene epoxide hydrolase, this strategy, which drastically reduces the search space and thus the screening effort, resulted in R,R- and S,S-selective mutants for the hydrolytic desymmetrization of cyclohexene oxide and other epoxides. X-ray crystal structures and docking studies of the mutants unveiled the source of stereoselectivity and shed light on the mechanistic intricacies of this enzyme.

PubMed: 25891639
DOI: 10.1002/anie.201501809

実験手法

X-RAY DIFFRACTION (2.3 Å)