Semi-rational engineering membrane binding domain of L-amino acid deaminase from Proteus vulgaris for enhanced α-ketoisocaproate.

ABSTRACT: α-Keto acids are important raw materials for pharmaceuticals and functional foods, which could be produced from cheap feed stock by whole cell biocatalysts containing L-amino acid deaminases (L-AADs). However, the production capacity is limited by the low activity of L-AADs. The L-AAD mediated redox reaction employs the electron transport chain to transfer electrons from the reduced FADH₂ to O₂, implying that the interaction between L-AAD and the cell membrane affects its catalytic activity. To improve the catalytic activity of L-AAD from Proteus vulgaris, we redesigned the membrane-bound hydrophobic insertion sequences (INS, residues 325-375) by saturation mutagenesis and high-throughput screening. Mutants D340N and L363N exhibited higher affinity and catalyti

SUBMITTER: Song Y 

PROVIDER: S-EPMC9561763 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature