Structure-Based Engineering of an Artificially Generated NADP⁺-Dependent d-Amino Acid Dehydrogenase.

ABSTRACT: A stable NADP⁺-dependent d-amino acid dehydrogenase (DAADH) was recently created from Ureibacillus thermosphaericusmeso-diaminopimelate dehydrogenase through site-directed mutagenesis. To produce a novel DAADH mutant with different substrate specificity, the crystal structure of apo-DAADH was determined at a resolution of 1.78 Å, and the amino acid residues responsible for the substrate specificity were evaluated using additional site-directed mutagenesis. By introducing a single D94A mutation, the enzyme's substrate specificity was dramatically altered; the mutant utilized d-phenylalanine as the most preferable substrate for oxidative deamination and had a specific activity of 5.33 μmol/min/mg at 50°C, which was 54-fold higher than that of the parent DAADH. In add

SUBMITTER: Hayashi J 

PROVIDER: S-EPMC5440713 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature