AlphaFold-Guided Semi-Rational Engineering of an (<i>R</i>)-Amine Transaminase for Green Synthesis of Chiral Amines.

ABSTRACT: Chiral amines are vital structural motifs in pharmaceuticals and agrochemicals, where enantiomeric purity governs bioactivity and environmental behavior. We identified a novel (R)-selective amine transaminase (MwoAT) from Mycobacterium sp. via genome mining, which exhibits activity toward the synthesis of the chiral amine (R)-1-methyl-3-phenylpropylamine. The enzyme displayed optimal activity at pH 7.0 and 40 °C, with high thermostability and solvent tolerance. Using an AlphaFold3-guided semi-rational engineering strategy integrating molecular docking, alanine scanning, and saturation mutagenesis, residue L175 was pinpointed as critical for substrate binding. The resulting L175G variant exhibited a 2.1-fold increase in catalytic efficiency (k_cat/K_m

SUBMITTER: Yang X 

PROVIDER: S-EPMC12564524 | biostudies-literature | 2025 Oct

REPOSITORIES: biostudies-literature