Rational design of the beta-galactosidase from Aspergillus oryzae to improve galactooligosaccharide production

The β-galactosidase from Aspergillus oryzae produces galactooligosaccharides with beneficial prebiotic effects through the transglycosylation of lactose. However, its low galactooligosaccharide yield greatly limits its use in industrial applications. Rational design and site-directed mutagenesis were used to produce three mutants of A. oryzae β-galactosidase: N140C, W806F, and N140C/W806F. The galactooligosaccharide yields of N140C (50.7%), W806F (49.3%), and N140C/W806F (59.8%) represent substantial improvements over that of the wild-type (35.7%). The galactooligosaccharide yield of N140C/W806F is the highest reported to date. Our rational design approach suggests novel strategies for further study of the β-galactosidase reaction mechanism and has produced mutants may be more useful in industrial applications than wild-type A. oryzae β-galactosidase.