Awakening the Sleeping Carboxylase Function of Enzymes: Engineering the Natural CO₂-Binding Potential of Reductases.

ABSTRACT: Developing new carbon dioxide (CO₂) fixing enzymes is a prerequisite to create new biocatalysts for diverse applications in chemistry, biotechnology and synthetic biology. Here we used bioinformatics to identify a "sleeping carboxylase function" in the superfamily of medium-chain dehydrogenases/reductases (MDR), i.e. enzymes that possess a low carboxylation side activity next to their original enzyme reaction. We show that propionyl-CoA synthase from Erythrobacter sp. NAP1, as well as an acrylyl-CoA reductase from Nitrosopumilus maritimus possess carboxylation yields of 3 ± 1 and 4.5 ± 0.9%. We use rational design to engineer these enzymes further into carboxylases by increasing interactions of the proteins with CO₂ and suppressing diffusion of water to the active sit

SUBMITTER: Bernhardsgrutter I 

PROVIDER: S-EPMC6650136 | biostudies-literature | 2019 Jun

REPOSITORIES: biostudies-literature