One-step synthesis of alpha-Gal epitope and globotriose derivatives by an engineered alpha-galactosidase

Directed evolution of an alpha-galactosidase (Aga2) from Bifidobacterium breve 203 by random mutagenesis and subsequently by site-directed mutagenesis provided a mutant enzyme V564N that showed high alpha-transgalactosylation efficiency with unobserved hydrolysis towards transglycosylation products. Using this enzyme, a one-step reaction for the simultaneous synthesis of alpha-Gal epitope and globotriose derivatives was achieved.