Using mutability landscapes of a promiscuous tautomerase to guide the engineering of enantioselective Michaelases

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. W. H., Quax, W. J. & Poelarends, G. J., 2016, In : Nature Communications. 7, 16 p., 10911.

Research output: Contribution to journalArticleAcademicpeer-review

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.

Original languageEnglish
Article number10911
Number of pages16
JournalNature Communications
Publication statusPublished - 2016
Related Press / Media
  1. Nature Communications paper: Klein enzym krijgt nieuwe functie

    Gerrit Poelarends


    1 item of Media coverage

    Press/Media: ResearchPopular

View all (1) »

Download statistics

No data available

ID: 30378952