2V3W

Crystal structure of the benzoylformate decarboxylase variant L461A from Pseudomonas putida

2V3W の概要

• エントリーDOI: 10.2210/pdb2v3w/pdb
• 関連するPDBエントリー: 1BFD 1MCZ 1PI3 1PO7 1Q6Z 1YNO
• 分子名称: BENZOYLFORMATE DECARBOXYLASE, MAGNESIUM ION, SULFATE ION, ... (5 entities in total)
• 機能のキーワード: lyase, calcium, magnesium, flavoprotein, thiamine pyrophosphate, rational protein design, aromatic hydrocarbons catabolism, thdp-dependent, mandelate pathway, metal-binding, decarboxylase, carboligation
• 由来する生物種: PSEUDOMONAS PUTIDA
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 227674.01

構造登録者

Gocke, D.,Walter, L.,Gauchenova, K.,Kolter, G.,Knoll, M.,Berthold, C.L.,Schneider, G.,Pleiss, J.,Mueller, M.,Pohl, M. (登録日: 2007-06-25, 公開日: 2008-01-22, 最終更新日: 2023-12-13)

主引用文献

Gocke, D.,Walter, L.,Gauchenova, K.,Kolter, G.,Knoll, M.,Berthold, C.L.,Schneider, G.,Pleiss, J.,Muller, M.,Pohl, M.
Rational Protein Design of Thdp-Dependent Enzymes-Engineering Stereoselectivity.
Chembiochem, 9:406-, 2008

PubMed Abstract: Benzoylformate decarboxylase (BFD) from Pseudomonas putida is an exceptional thiamin diphosphate-dependent enzyme, as it catalyzes the formation of (S)-2-hydroxy-1-phenylpropan-1-one from benzaldehyde and acetaldehyde. This is the only currently known S-selective reaction (92 % ee) catalyzed by this otherwise R-selective class of enzymes. Here we describe the molecular basis of the introduction of S selectivity into ThDP-dependent decarboxylases. By shaping the active site of BFD through the use of rational protein design, structural analysis, and molecular modeling, optimal steric stabilization of the acceptor aldehyde in a structural element called the S pocket was identified as the predominant interaction for adjusting stereoselectivity. Our studies revealed Leu461 as a hot spot for stereoselectivity in BFD. Exchange to alanine and glycine resulted in variants that catalyze the S-stereoselective addition of larger acceptor aldehydes, such as propanal with benzaldehyde and its derivatives-a reaction not catalyzed by the wild-type enzyme. Crystal structure analysis of the variant BFDL461A supports the modeling studies.

PubMed: 18224647
DOI: 10.1002/CBIC.200700598

実験手法

X-RAY DIFFRACTION (2.2 Å)