Abstract
A β-galactosidase (β-GalINF) was directly isolated from feces of an 8-month-old infant. Mass spectrum analysis showed β-GalINF with coverage over 50% to the β-galactosidase from Bifidobacterium longum EK3. Accordingly, the β-galINF was amplified from the feces metagenomic DNA by degenerate primers. After overexpressed in Escherichia coli, the β-GalINF was purified and biochemical characterized. β-GalINF existed as homotetramer and homodimer, whose activity (optimal at 50 °C, pH 6.5) was exhilaratingly increased to 484% by artificial intestinal juice. The Km and Vmax values for oNPG and lactose were 20.95 ± 2.76 mM, 5004.50 ± 318.8 μmol min−1 mg−1 and 140.2 ± 17.7 mM, 293.1 ± 14.7 μmol min−1 mg−1, respectively. The production rate of galacto-oligosaccharides by β-GalINF from 20% lactose at 50 °C was 33.4 ± 0.67%. These results suggested the β-GalINF with high hydrolytic and transgalactosylation activity from the infant intestinal has great potential as infant lactase preparation. Moreover, this study provided a new way for exploring undetected enzymes by uncultured-dependent methods.
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Funding
This work was supported by grants from the National Key Research and Development Program of China (2017YFD0400300), the National Natural Science Foundation of China (31871767), and the Public Service Sectors (Agriculture) Special and Scientific Research Projects (201503134).
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Xin, Y., Guo, T., Zhang, Y. et al. A new β-galactosidase extracted from the infant feces with high hydrolytic and transgalactosylation activity. Appl Microbiol Biotechnol 103, 8439–8448 (2019). https://doi.org/10.1007/s00253-019-10092-x
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DOI: https://doi.org/10.1007/s00253-019-10092-x