3ZVH

Methylaspartate ammonia lyase from Clostridium tetanomorphum mutant Q73A

Summary for 3ZVH

• Entry DOI: 10.2210/pdb3zvh/pdb
• Related: 1KCZ 1KD0 3ZVI
• Descriptor: METHYLASPARTATE AMMONIA-LYASE, MAGNESIUM ION, GLYCEROL, ... (6 entities in total)
• Functional Keywords: lyase, enolase
• Biological source: CLOSTRIDIUM TETANOMORPHUM; More
• Total number of polymer chains: 2
• Total formula weight: 97475.06

Authors

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. (deposition date: 2011-07-25, release date: 2012-05-02, Last modification date: 2023-12-20)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (1.99 Å)