Purification and characterization of a two-peptide bacteriocin with synergistic activity from Enterococcus faecalis MGL-3
Rodney H. Perez
Abstract:
Bacteriocins are food-grade bioactive peptides produced by some
bacterial strains, including lactic acid bacteria (LAB). Bacteriocins
derived from food-grade LAB possess enormous potential for various
applications in the food and medical industries. A newly isolated lactic
acid bacterium from a fermented vegetable, strain MGL-3, was found
to produce two bioactive peptides. In this study, the identification of
strain MGL-3 as well as the purification and partial characterization of
its bacteriocinogenic peptides are described. Morphological,
biochemical, and bioinformatic analyses revealed that strain MGL-3 is
an Enterococcus faecalis strain. Two bioactive peptides were purified
from the cell-free supernatant of strain MGL-3 after a 3-step
purification scheme involving salt precipitation and chromatographic
techniques. Characterization of these peptides showed that they
exhibit relatively weak bioactivity when tested alone but manifest very
potent bioactivity when combined, suggesting synergistic activity.
The antibacterial spectrum of these peptides was found to target
many bacterial strains that are known contaminants in the vegetable
pickle industry, highlighting their potential utility in the industry.
Nonetheless, further study is needed to confirm the novelty of these
peptides, such as the determination of their amino acid sequence
and/or the identification of their biosynthetic gene cluster.
References:
- Chen, H., & Hoover, D. G. (2003). Bacteriocins and their food applications. Comprehensive Reviews in Food Science and Food Safety, 2(3), 82–100. https://doi.org/10.1111/j.1541-4337.2003.tb00016.x
- Cleveland, J., Montville, T. J., Nes, I. F., & Chikindas, M. L. (2001). Bacteriocins: Safe, natural antimicrobials for food preservation. International Journal of Food Microbiology, 71(1), 1–20. http://www.ncbi.nlm.nih.gov/pubmed/11764886
- Cotter, P. D., Deegan, L. H., Lawton, E. M., Draper, L. A., O’Connor, P. M., Hill, C., & Ross, R. P. (2006). Complete alanine scanning of the two-component lantibiotic lacticin 3147: Generating a blueprint for rational drug design. Molecular Microbiology, 62(3), 735–747. https://doi.org/10.1111/j.1365-2958.2006.05398.x
- Cotter, P. D., Hill, C., & Ross, R. P. (2005). Bacteriocins: Developing innate immunity for food. Nature Reviews Microbiology, 3(10), 777–788. https://doi.org/10.1038/nrmicro1273
- Hammami, R., Zouhir, A., Le Lay, C., Ben Hamida, J., & Fliss, I. (2010). BACTIBASE second release: A database and tool platform for bacteriocin characterization. BMC Microbiology, 10, Article 22. https://doi.org/10.1186/1471-2180-10-22
- Holzapfel, W. H., Geisen, R., & Schillinger, U. (1995). Biological preservation of foods with reference to protective cultures, bacteriocins and food-grade enzymes. International Journal of Food Microbiology, 24(3), 343–362. http://www.ncbi.nlm.nih.gov/pubmed/7710912
- Hu, C. B., Malaphan, W., Zendo, T., Nakayama, J., & Sonomoto, K. (2010). Enterocin X, a novel two-peptide bacteriocin from Enterococcus faecium KU-B5, has an antibacterial spectrum entirely different from those of its component peptides. Applied and Environmental Microbiology, 76(13), 4542–4545. https://doi.org/10.1128/AEM.02264-09
- Klaenhammer, T. R. (1993). Genetics of bacteriocins produced by lactic acid bacteria. FEMS Microbiology Reviews, 12(1), 39–85. http://www.ncbi.nlm.nih.gov/pubmed/8398217
- Kumar, S., Stecher, G., Suleski, M., Sanderford, M., Sharma, S., & Tamura, K. (2024).
MEGA12: Molecular Evolutionary Genetics Analysis version 12 for adaptive and green computing. Molecular Biology and Evolution, 41(12),1–9. https://academic.oup.com/mbe/article/41/12/msae263/7930299
- Li, K., Lin, K., Li, Z., Zhang, Q., Song, F., Che, Z., Chen, G., & Xiang, W. (2014).
Spoilage and pathogenic bacteria associated with spoilage process of Sichuan pickle during the spontaneous fermentation. Food Science and Technology Research, 20(4), 899–904. https://doi.org/10.3136/fstr.20.899
- Masuda, Y., Zendo, T., Sawa, N., Perez, R. H., Nakayama, J., & Sonomoto, K. (2012).
Characterization and identification of weissellicin Y and weissellicin M, novel bacteriocins produced by Weissella hellenica QU 13. Journal of Applied Microbiology, 112(1), 99–108. https://doi.org/10.1111/j.1365-2672.2011.05180.x
- Moll, G., Ubbink-Kok, T., Hildeng-Hauge, H., Nissen-Meyer, J., Nes, I. F., Konings, W. N., & Driessen, A. J. (1996). Lactococcin G is a potassium ion–conducting, two-component bacteriocin. Journal of Bacteriology, 178(3), 600–605. http://www.ncbi.nlm.nih.gov/pubmed/8550488
- Nissen-Meyer, J., Holo, H., Havarstein, L. S., Sletten, K., & Nes, I. F. (1992).
A novel lactococcal bacteriocin whose activity depends on the complementary action of two peptides. Journal of Bacteriology, 174(17), 5686–5692. http://www.ncbi.nlm.nih.gov/pubmed/1512201
- Nissen-Meyer, J., Oppegard, C., Rogne, P., Haugen, H. S., & Kristiansen, P. E. (2010).
Structure and mode of action of the two-peptide (class IIb) bacteriocins. Probiotics and Antimicrobial Proteins, 2(1), 52–60. https://doi.org/10.1007/s12602-009-9021-z
- Oppegård, C., Rogne, P., Emanuelsen, L., Kristiansen, P. E., Fimland, G., & Nissen-Meyer, J. (2007).
The two-peptide class II bacteriocins: Structure, production, and mode of action. Journal of Molecular Microbiology and Biotechnology, 13(4), 210–219. https://doi.org/10.1159/000104750
- Perez, R. H., Himeno, K., Zendo, T., & Sonomoto, K. (2020).
Critical fermentation factors that influence the production of multiple bacteriocins of Enterococcus faecium NKR-5-3. Annals of Tropical Research, 42(2), 71–84. https://doi.org/10.32945/atr4226.2020
- Perez, R. H., Zendo, T., & Sonomoto, K. (2014).
Novel bacteriocins from lactic acid bacteria (LAB): Various structures and applications. Microbial Cell Factories, 13(Suppl 3). https://doi.org/10.1186/1475-2859-13-S1-S3
- Sawa, N., Zendo, T., Kiyofuji, J., Fujita, K., Himeno, K., Nakayama, J., & Sonomoto, K. (2009).
Identification and characterization of lactocyclicin Q, a novel cyclic bacteriocin produced by Lactococcus sp. strain QU 12. Applied and Environmental Microbiology, 75(6), 1552–1558. https://doi.org/10.1128/AEM.02299-08
- Tajima, K., Arai, S., Ogata, K., Nagamine, T., Matsui, H., Nakamura, M., Aminov, R. I., & Benno, Y. (2000).
Rumen bacterial community transition during adaptation to a high-grain diet. Anaerobe, 6(5), 273–284. https://doi.org/10.1006/anae.2000.0353
- U.S. Food and Drug Administration. (1988).
Nisin preparation: Affirmation of GRAS status as a direct human food ingredient (21 CFR Part 184). Federal Register, 53, 11247–11251. https://archives.federalregister.gov/issue_slice/1988/4/6/11245-11253.pdf
- Willey, J. M., & van der Donk, W. A. (2007).
Lantibiotics: Peptides of diverse structure and function. Annual Review of Microbiology, 61, 477–501. https://doi.org/10.1146/annurev.micro.61.080706.093501
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