Premium
Characterization of Carboxylic Acid Reductases as Enzymes in the Toolbox for Synthetic Chemistry
Author(s) -
Finnigan William,
Thomas Adam,
Cromar Holly,
Gough Ben,
Snajdrova Radka,
Adams Joseph P.,
Littlechild Jennifer A.,
Harmer Nicholas J.
Publication year - 2017
Publication title -
chemcatchem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.497
H-Index - 106
eISSN - 1867-3899
pISSN - 1867-3880
DOI - 10.1002/cctc.201601249
Subject(s) - adenylylation , enzyme , chemistry , substrate (aquarium) , biocatalysis , stereochemistry , carboxylic acid , pyrophosphate , enzyme catalysis , combinatorial chemistry , cofactor , biochemistry , biosynthesis , reaction mechanism , catalysis , biology , ecology
Abstract Carboxylic acid reductase enzymes (CARs) meet the demand in synthetic chemistry for a green and regiospecific route to aldehydes from their respective carboxylic acids. However, relatively few of these enzymes have been characterized. A sequence alignment with members of the ANL (Acyl‐CoA synthetase/ NRPS adenylation domain/Luciferase) superfamily of enzymes shed light on CAR functional dynamics. Four unstudied enzymes were selected by using a phylogenetic analysis of known and hypothetical CARs, and for the first time, a thorough biochemical characterization was performed. Kinetic analysis of these enzymes with various substrates shows that they have a broad but similar substrate specificity. Electron‐rich acids are favored, which suggests that the first step in the proposed reaction mechanism, attack by the carboxylate on the α‐phosphate of adenosine triphosphate (ATP), is the step that determines the substrate specificity and reaction kinetics. The effects of pH and temperature provide a clear operational window for the use of these CARs, whereas an investigation of product inhibition by NADP + , adenosine monophosphate, and pyrophosphate indicates that the binding of substrates at the adenylation domain is ordered with ATP binding first. This study consolidates CARs as important and exciting enzymes in the toolbox for sustainable chemistry and provides specifications for their use as a biocatalyst.