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The potential of the endolysin Lysdb from Lactobacillus delbrueckii phage for combating Staphylococcus aureus during cheese manufacture from raw milk

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Abstract

Phage endolysins have received increased attention in recent times as potential antibacterial agents and the biopreservatives in food production processes. Staphylococcus aureus is one of the most common pathogens in bacterial food poisoning outbreaks. In this study, the endolysin Lysdb, one of the two-component cell lysis cassette of Lactobacillus delbrueckii phage phiLdb, was shown to possess a muramidase domain and catalytic sites with homology to Chalaropsis-type lysozymes. Peptidoglycan hydrolytic bond specificity determination revealed that Lysdb was able to cleave the 6-O-acetylated peptidoglycans present in the cell walls of S. aureus. Turbidity reduction assays demonstrated that Lysdb could effectively lyse the S. aureus live cells under acidic and mesothermal conditions. To further evaluate the ability of Lysdb as a potential antibacterial agent against S. aureus in cheese manufacture, Lactobacillus casei BL23 was engineered to constitutively deliver active Lysdb to challenge S. aureus in lab-scale cheese making from raw milk. Compared with the raw milk, the viable counts of S. aureus were reduced by 105-fold in the cheese inoculated with the engineered L. casei strain during the fermentation process, and the pathogenic bacterial numbers remained at a low level (104 CFU/g) after 6 weeks of ripening at 10 °C. Taken together, all results indicated that the Lysdb has the function as an effective tool for combating S. aureus during cheese manufacture from raw milk.

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Acknowledgments

We would like to thank M. van de Guchte, S. Hazebrouck, and N. Galleron for the kindness of providing L. delbrueckii subsp. bulgaricus ATCC11842, L. casei BL23, and pSec:Nuc vector, respectively. This work was supported by grants from the National Natural Science Foundation of China (grant no. 31271905), Hi-tech Research and Development Program of China (no. 2011AA100902), and the Fundamental Research Funds of Shandong University (no. 2014GN024).

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  1. State Key Laboratory of Microbial Technology, Shandong University, 27 Shanda Nanlu, Jinan, 250100, People’s Republic of China

    Tingting Guo, YongPing Xin, Chenchen Zhang, Xudong Ouyang & Jian Kong

  2. School of Environmental Science and Engineering, Shandong University, Jinan, 250100, People’s Republic of China

    Tingting Guo

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  1. Tingting Guo
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Guo, T., Xin, Y., Zhang, C. et al. The potential of the endolysin Lysdb from Lactobacillus delbrueckii phage for combating Staphylococcus aureus during cheese manufacture from raw milk. Appl Microbiol Biotechnol 100, 3545–3554 (2016). https://doi.org/10.1007/s00253-015-7185-x

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