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Surface Display of Heterologous β-Galactosidase in Food-Grade Recombinant Lactococcus lactis

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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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Acknowledgements

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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Authors and Affiliations

  1. Department of Microbiology, Third Military Medical University (Army Medical University), Chongqing, China

    Supeng Yin, Hongbin Zhu, Mengyu Shen, Gang Li, Shuguang Lu, Yan Zhao, Shuai Le, Yinling Tan, Fuquan Hu & Jing Wang

  2. State Key Laboratory of Trauma, Burns, and Combined Injury, Institute of Burn Research, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, China

    Supeng Yin & Yizhi Peng

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  1. Supeng Yin
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Correspondence to Jing Wang.

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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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