3ZVI

Methylaspartate ammonia lyase from Clostridium tetanomorphum mutant L384A

3ZVI の概要

• エントリーDOI: 10.2210/pdb3zvi/pdb
• 関連するPDBエントリー: 1KCZ 1KD0 3ZVH
• 分子名称: METHYLASPARTATE AMMONIA-LYASE, GLYCEROL, PROPIONAMIDE, ... (6 entities in total)
• 機能のキーワード: lyase, enolase
• 由来する生物種: CLOSTRIDIUM TETANOMORPHUM
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 97587.20

構造登録者

Raj, H.,Szymanski, W.,de Villiers, J.,Rozeboom, H.J.,Veetil, V.P.,Reis, C.R.,de Villiers, M.,de Wildeman, S.,Dekker, F.J.,Quax, W.J.,Thunnissen, A.M.W.H.,Feringa, B.L.,Janssen, D.B.,Poelarends, G.J. (登録日: 2011-07-25, 公開日: 2012-05-02, 最終更新日: 2024-11-13)

主引用文献

Raj, H.,Szymanski, W.,De Villiers, J.,Rozeboom, H.J.,Veetil, V.P.,Reis, C.R.,De Villiers, M.,Dekker, F.J.,De Wildeman, S.,Quax, W.J.,Thunnissen, A.M.W.H.,Feringa, B.L.,Janssen, D.B.,Poelarends, G.J.
Engineering Methylaspartate Ammonia Lyase for the Asymmetric Synthesis of Unnatural Amino Acids.
Nat.Chem., 4:478-, 2012

PubMed Abstract: The redesign of enzymes to produce catalysts for a predefined transformation remains a major challenge in protein engineering. Here, we describe the structure-based engineering of methylaspartate ammonia lyase (which in nature catalyses the conversion of 3-methylaspartate to ammonia and 2-methylfumarate) to accept a variety of substituted amines and fumarates and catalyse the asymmetric synthesis of aspartic acid derivatives. We obtained two single-active-site mutants, one exhibiting a wide nucleophile scope including structurally diverse linear and cyclic alkylamines and one with broad electrophile scope including fumarate derivatives with alkyl, aryl, alkoxy, aryloxy, alkylthio and arylthio substituents at the C2 position. Both mutants have an enlarged active site that accommodates the new substrates while retaining the high stereo- and regioselectivity of the wild-type enzyme. As an example, we demonstrate a highly enantio- and diastereoselective synthesis of threo-3-benzyloxyaspartate (an important inhibitor of neuronal excitatory glutamate transporters in the brain).

PubMed: 22614383
DOI: 10.1038/NCHEM.1338

実験手法

X-RAY DIFFRACTION (1.9 Å)