Engineering an acetoacetyl-CoA reductase from Cupriavidus necator toward NADH preference under physiological conditions.

ABSTRACT: The coupling of PHB generation with NADH reoxidation is required to generate PHB as a fermentation product. A fundamental trait to accomplish this feature is to express a functional NADH-preferring acetoacetyl-CoA reductase, engaged in PHB accumulation. One way to obtain such a reductase is by engineering the cofactor preference of the acetoacetyl-CoA reductase encoded by the phaB1 gene from Cupriavidus necator (AAR^Cn1). Aiming to have a deeper understanding of the structural determinants of the cofactor preference in AAR^Cn1, and to obtain an NADH-preferring acetoacetyl-CoA reductase derived from this protein, some engineered enzymes were expressed, purified and kinetically characterized, together with the parental AAR^Cn1. One of these engineered enzymes, C

SUBMITTER: Olavarria K 

PROVIDER: S-EPMC8904767 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature