5CLO

Crystal structure of a 4-oxalocrotonate tautomerase mutant in complex with nitrostyrene at 2.3 Angstrom

5CLO の概要

• エントリーDOI: 10.2210/pdb5clo/pdb
• 分子名称: 2-hydroxymuconate tautomerase, trans beta nitrostyrene (3 entities in total)
• 機能のキーワード: 4-oxalocrotonate tautomerase, beta-alpha-beta structural motif, tautomerase superfamily, isomerase
• 由来する生物種: Pseudomonas putida
• タンパク質・核酸の鎖数: 18
• 化学式量合計: 114704.78

構造登録者

Thunnissen, A.M.W.H.,Poddar, H. (登録日: 2015-07-16, 公開日: 2016-03-09, 最終更新日: 2024-01-10)

主引用文献

van der Meer, J.Y.,Poddar, H.,Baas, B.J.,Miao, Y.,Rahimi, M.,Kunzendorf, A.,van Merkerk, R.,Tepper, P.G.,Geertsema, E.M.,Thunnissen, A.M.,Quax, W.J.,Poelarends, G.J.
Using mutability landscapes of a promiscuous tautomerase to guide the engineering of enantioselective Michaelases.
Nat Commun, 7:10911-10911, 2016

PubMed Abstract: The Michael-type addition reaction is widely used in organic synthesis for carbon-carbon bond formation. However, biocatalytic methodologies for this type of reaction are scarce, which is related to the fact that enzymes naturally catalysing carbon-carbon bond-forming Michael-type additions are rare. A promising template to develop new biocatalysts for carbon-carbon bond formation is the enzyme 4-oxalocrotonate tautomerase, which exhibits promiscuous Michael-type addition activity. Here we present mutability landscapes for the expression, tautomerase and Michael-type addition activities, and enantioselectivity of 4-oxalocrotonate tautomerase. These maps of neutral, beneficial and detrimental amino acids for each residue position and enzyme property provide detailed insight into sequence-function relationships. This offers exciting opportunities for enzyme engineering, which is illustrated by the redesign of 4-oxalocrotonate tautomerase into two enantiocomplementary 'Michaelases'. These 'Michaelases' catalyse the asymmetric addition of acetaldehyde to various nitroolefins, providing access to both enantiomers of γ-nitroaldehydes, which are important precursors for pharmaceutically active γ-aminobutyric acid derivatives.

PubMed: 26952338
DOI: 10.1038/ncomms10911

実験手法

X-RAY DIFFRACTION (2.3 Å)