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.
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This research was supported by National Natural Science Foundation of China (31900035) and Central Public-interest Scientific Institution Basal Research Fund, CAFS (NO.2020TD67).
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JS, CY and JH contributed to the study conception and design. Data collection and analysis were performed by JS, CY and YY. Material preparation was performed by YL. The first draft of the manuscript was written by JS and WW. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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Sun, J., Yao, C., Li, Y. et al. A novel salt-tolerant GH42 β-galactosidase with transglycosylation activity from deep-sea metagenome. World J Microbiol Biotechnol 38, 154 (2022). https://doi.org/10.1007/s11274-022-03348-8
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DOI: https://doi.org/10.1007/s11274-022-03348-8