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

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-fu

SUBMITTER: van der Meer JY 

PROVIDER: S-EPMC4786785 | biostudies-literature | 2016 Mar

REPOSITORIES: biostudies-literature