A Novel Salt-tolerant GH42 β-galactosidase With Transglycosylation ...

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Abstract

β-Galactosidase is a widely adopted enzyme in the food and pharmaceutical industries. Metagenome techniques have the advantage of discovering novel functional genes, particularly potential genes from uncultivated microbes. In this study, a novel GH42 β-galactosidase isolated from a deep-sea metagenome was overexpressed in Escherichia coli BL21 (DE3) and purified by affinity chromatography. The optimal temperatures and pH of the enzyme for o-nitrophenyl-β-D-galactopyranoside (oNPG) and lactose were 40 ℃, 6.5 and 50 ℃, 7, respectively. The enzyme was stable at temperatures between 4 and 30 ℃ and within the pH range of 6-9. Moreover, it was highly tolerant to salt and inhibited by Zn2+ and Cu2+. The kinetic values of Km and kcat of the enzyme against oNPG were 1.1 mM and 57.8 s-1, respectively. Furthermore, it showed hydrolysis and transglycosylation activity to lactose and the extra monosaccharides could improve the productivity of oligosaccharides. Overall, this recombinant β-galactosidase is a potential biocatalyst for the hydrolysis of milk lactose and synthesis of functional oligosaccharides.

Keywords: Lactose; Metagenome; Salt tolerance; Transglycosylation; β-Galactosidase.

© 2022. The Author(s), under exclusive licence to Springer Nature B.V.

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References

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• Enzyme Stability Actions
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• Escherichia coli / genetics Actions
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Grants and funding

• 31900035/National Natural Science Foundation of China
• NO.2020TD67/Central Public-interest Scientific Institution Basal Research Fund, Chinese Academy of Fishery Sciences

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