The invention belongs to the technical field of
enzyme engineering, and particularly relates to a method for enhancing alpha-galactosidase
oligosaccharide synthesis capacity through site-specific
mutagenesis and an engineered
enzyme. The
mutant enzyme K480R for enhancing the
oligosaccharide synthesis capability of the alpha-galactosidase is obtained by carrying out site-
directed mutagenesis on the alpha-galactosidase from
lactobacillus amyloliquefaciens L6 by utilizing
a site-
directed mutagenesis technology, has efficient and accurate catalytic capability, can efficiently convert melibiose into
oligosaccharide, has the conversion rate of 34.30%, and can be used for preparing a
mutant enzyme K480R for enhancing the oligosaccharide synthesis capability of the alpha-galactosidase. The activity of the alpha-galactosidase is higher than that of alpha-galactosidase derived from
bacteria reported in China, side reactions are greatly reduced, and the production efficiency is improved. The synthesized oligosaccharide provides a new effective component for developing novel functional foods and medical products in the food field and the
medical health care aspect. Therefore, the
mutant enzyme has application potential in multiple industries such as food,
medicine,
cosmetics and the like, industrial innovation can be promoted, and considerable economic and
social benefits can be created.