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Food Chem. 2019 Jul 15;286:362-367. doi: 10.1016/j.foodchem.2019.01.212. Epub 2019 Feb 10.

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

Author information

1
State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China; School of Biotechnology and Key Laboratory of Industrial Biotechnology Ministry of Education, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China.
2
State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China; School of Biotechnology and Key Laboratory of Industrial Biotechnology Ministry of Education, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China; International Joint Laboratory on Food Safety, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China. Electronic address: jingwu@jiangnan.edu.cn.
3
State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China; School of Biotechnology and Key Laboratory of Industrial Biotechnology Ministry of Education, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China; International Joint Laboratory on Food Safety, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China. Electronic address: wudan@jiangnan.edu.cn.

Abstract

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.

KEYWORDS:

Galactooligosaccharide; Glycoside hydrolases; Rational design; Transglycosylation; β-Galactosidase

PMID:
30827619
DOI:
10.1016/j.foodchem.2019.01.212
[Indexed for MEDLINE]

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