Abstract
β-Galactosidase is an essential enzyme for the metabolism of lactose in human beings and has an important role in the treatment of lactose intolerance (LI). β-Galactosidase expressed by intestinal microflora, such as lactic acid bacteria (LAB), also alleviates LI. A promising approach to LI management is to exploit a food-grade LAB delivery system that can inhabit the human intestine and overproduce β-galactosidase. In this study, we constructed a food-grade β-galactosidase surface display delivery system and then integrated into the chromosome of Lactococcus lactis (L. lactis) NZ9000 using recombination. Western blot and immunofluorescence analyses confirmed that β-galactosidase was expressed on the cell surface of recombinant L. lactis stain NZ-SDL. The whole-cell biocatalyst exhibits Vmax and Km values of 121.38 ± 7.17 UONPG/g and 65.36 ± 5.54 mM, based on ONPG hydrolysis. The optimum temperature for enzyme activity is 37 °C and the optimum pH is 5.0. Activity of the whole-cell biocatalyst is promoted by Mg2+, Ca2+, and K+, but inhibited by Zn2+, Fe2+, and Fe3+. The system has a thermal stability similar to purified β-galactosidase but better pH stability, and is also more stable in artificial intestinal juice. Oral administration and intraperitoneal injections of NZ-SDL in mice cause no detectable health effects. In conclusion, we have successfully constructed a food-grade gene expression system in L. lactis that displays β-galactosidase on the cell surface. This system exhibits good enzyme activity and stability in vitro, and is safe in vivo. It is therefore a promising candidate for use in LI management.
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This study was funded by National Natural Science Foundation of China (NSFC, Grant No. 31201341), and Third Military Medical University Youth Science Foundation (Grant No. 2010XQN06).
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The protocol for animal experiments was approved by the Laboratory Animal Welfare and Ethics Committee of Third Military Medical University (Army Medical University).
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Yin, S., Zhu, H., Shen, M. et al. Surface Display of Heterologous β-Galactosidase in Food-Grade Recombinant Lactococcus lactis. Curr Microbiol 75, 1362–1371 (2018). https://doi.org/10.1007/s00284-018-1531-z
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DOI: https://doi.org/10.1007/s00284-018-1531-z