29Aug 2017

PRODUCTION OF A NISIN-LIKE BACTERIOCIN FROM Lactococcus lactis subsp. lactis STRAIN CCSU 1011 ISOLATED FROM MILK.

  • Prof. and Head, Department of Microbiology, Shree Bankey Bihari Dental College & Research Centre, Ghaziabad, (UP) India.
Crossref Cited-by Linking logo

  • Abstract
  • Keywords
  • References
  • Cite This Article as
  • Corresponding Author

A total of 104 milk samples were collected from different dairies in Meerut region from which 120 isolates of lactic acid bacteria (LAB) were isolated. Of these, 68 strains showed antibacterial activity against the sensitive test organism Lactococcus lactis subsp. lactis MTCC 3038. The isolate which showed strongest activity was identified as Lactococcus lactis subsp. lactis using the characters of cell morphology, biochemical tests and carbohydrate fermentation profile. It was given the strain number as Lactococcus lactis subsp. lactis CCSU 1011. It produced bacteriocin which was active against closely related lactic acid bacteria as well as a wide range of food pathogens including Salmonella typhi, Listeria monocytogenes, Clostridium perfringens, Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis, Shigella flexineri, Bacillus cereus, and Enterococcus faecalis. The antimicrobial spectrum was nearly identical to that of nisin. Optimum bacteriocin was produced at the initial pH of 7.0 and at 35 ?C temperature after 24 h of incubation period in MRS broth with 1.5% Tween-80. Bacteriocin activity was not destroyed by exposure to elevated temperatures at low pH values, but the activity was lost at high pH values. This bacteriocin was inactivated by proteases and pronases but not by trypsin, pepsin and α- amylase. The bacteriocin was purified using ammonium sulphate precipitation, cation exchange chromatography (CM-cellulose) and gel filtration (Sephadex G-50). The apparent molecular weight of the purified bacteriocin was calibrated as approximately 3.5 kDa on SDS-PAGE similar to that of nisin. The bacteriocin exhibited bactericidal mode of action against Lactococcus lactis subsp. lactis MTCC 3038. These results indicate that bacteriocin produced by Lactococcus lactis subsp. lactis CCSU 1011 is a nisin or nisin-like bacteriocin.

  1. Ariyapitipum, T. Mustapha, A. and Clarke, A.D. (1999): Microbial shelf life determinartion of vacuum- packaged fresh beef treated with polylactic acid, lactic acid, and nisin solutions. J Food Prot, 62: 913-920.
  2. Batish, V.K., Ram, L. and Grover, S. (1990): Studies on environmental and nutritional factors on production of antifungal substance by nutritional factors on production of antifungal substance by Lactobacillus acidophilus Food Microbiol., 7: 199-206.
  3. Bolotin, A.1, Wincker, P., Mauger, S., Jaillon, O., Malarme, K., Weissenbach, J., Ehrlich, S.D., Sorokin, A.(2001): The complete genome sequence of the lactic acid bacterium Lactococcus lactis ssp. lactis IL1403. Genome Res. 2001 May;11(5):731-53.
  4. Chang, I.S., Kim, B.H. and Shin, P..K (1997): Use of sulphate and hydrogen peroxide to control bacterial contamination in ethanol production. Appl Environ Microbiol. 63: 1-6.
  5. Choi, Y.O. and Ahn, C. (1997): Plasmid-associated bacteriocin production by Leuconostoc LAB 145-3A isolated from kimchi. J Microbiol Biotech 7: 409-416.
  6. Choi, S.H., Oh, C.T., Kim, S.H., Kim, Y.T., Jeon, S.H. (2000):. Effects of Polycomb group mutations on the expression of Ultrabithorax in the Drosophila visceral mesoderm.? Cells 10(2): 156--161.
  7. Cleeveland, J., Mantiville, T. J., Ness, I. F. and Chiknids, M. L. (2001): Bacteriocins: safe antimicrobials for food preservation. International Journal of Food Microbiology 71,1-20.
  8. Daeschel, M.A., (1989): Antimicrobial substances from acid lactic bacteria for use as food preservative. Food Technology, 43,164-167.
  9. De Vuys,t L. and Vandamme,.. E J (1994a): Bacteriocins of Lactic Acid Bacteria: Microbiology, Genetics and Applications. Blackie Academic & Professional, London.
  10. De Vuyst, L. (1995): Nutritional factors affecting nisin production by Lactococcus lactis lactis NIZO 22186 in a synthetic medium. J. Appl. Bacteriol., 78, 28-33.
  11. Gross, E. and Morell, J. L. (1971): Structure of nisin. Am. Chem. Soc., 93, 4634-4635.
  12. Tagg, J.R. and ?McGiven, (1971): A.R. Assay System for Bacteriocins Appl Microbiol. 1971 May; 21(5): 943.
  13. Jack, R. W.; Tagg, J. R. and Ray, B. (1995): Bacteriocins of Gram-positive bacteria. Rev., 59, 171-200.
  14. Jay J.M (1982): Antimicrobial properties of diacetyl. Appl Environ Microbiol 44: 525-532.
  15. Khay, E., Idaomar, M., Castro, L.M.P., Bern?rdez, P.F., Senhaji, N.S. and Abrini, J. (2011): Antimicrobial activities of the bacteriocin-like substances produced by lactic acid bacteria isolated from Moroccan dromedary milk. African Journal of Biotechnology 10(51): 10447?10455.
  16. Klaenhammer, T. (1988): Bacteriocins of lactic acid bacteria. Biochimie 70:337-349.
  17. Ko, S.H. and Ahn, C. (2000): Bacteriocin production by Lactococcus lactis KCA2386 isolated from white kimchi. Food Sci. Biotechnol., 9: 263-269.
  18. Laemmli, U.K. (1970): Cleavage of structural proteins during the assembly of the head bacteriophage T4. Nature 27: 680-68.
  19. Loh, J.Y., Lim, Y.Y. and Ting, A.S.Y. (2017): Bacteriocin-like substances produced by Lactococcus lactissubsp. lactis CF4MRS isolated from fish intestine: Antimicrobial activities and inhibitory properties International Food Research Journal 24(1): 394-400 (February 2017).
  20. Muriana, P.M. (1996): Bacteriocins for control of Listeria spp. in food.Journal of Food Protection 1996 Supplement,54?63.
  21. Nagalakshmi ,P.K.,.Sumathi, R.,Kanimozhi ,K.,and Sivakumar,T. J .(2013): Isolation of Bacteriocin nisin producing Lactococcus lactis from dairy product .J Acad. Indus. Res.Vol. 1(10) March.
  22. Park, J.H., Schroeder, A.J., Helfrich-Forster, C., Jackson, F.R., Ewer, J. (2003): Targeted ablation of CCAP neuropeptide-containing neurons of Drosophila causes specific defects in execution and circadian timing of ecdysis behavior.? Development 130(12): 2645--2656.
  23. Ray, B., Miller, K.W. & Jain, M.K. (2001): Bacteriocins of Lactic Acid Bacteria: Current Perspective. Indian J Microbiol 41: 01-21.
  24. Schillinger, U. (1999): Isolation and identification of Lactobacilli from novel-type probiotic and mild yoghurts and their stability during refrigerated storage. Int J Food Microbiol 47: 79-87.
  25. Schillinger, U.; Becker, B.; Vignolo, G. and Holzapfel, W. H. (2001): Efficacy of nisin in combination with protective cultures against Listeria monocytogenes Scott A in tofu. J. Food Microbiol., 71, 159-168.
  26. Scott, S.C., Allen, J.G., & John, J.I. (1997): Purification and Characteriazation of Staphylococcin Bac R1, a broad-spectrum bacteriocin. Appl Environ Microbiol 63: 4185-4190.
  27. Tagg, J. R., Dajani, A. S. & Wannamaker, L. W. (1976): Bacteriocins of gram positive bacteria. Bacteriol Rev 40 722?756.
  28. Tagg, J.R. and McGiven, A.R. (1971): Assay system for bacteriocins..Applied and Environmental Microbiology21, 943?948.
  29. Villamil, L., Reyes, C. and Mart?nez-Silva, M.A. (2014): In vivo and in vitro assessment of Lactobacillus acidophilus as probiotic for tilapia (Oreochromis niloticus, Perciformes: Cichlidae) culture improvement. Aquaculture Research 45(7): 1116?1125.
  30. Yang, R., Johnson, M.C. & Ray, B. (1992): Novel method to extract large amounts of bacteriocin from lactic acid bacteria. Appl Environ Microbiol 58: 3355-3359.

[Mukta Sharma. (2017); PRODUCTION OF A NISIN-LIKE BACTERIOCIN FROM Lactococcus lactis subsp. lactis STRAIN CCSU 1011 ISOLATED FROM MILK. Int. J. of Adv. Res. 5 (Aug). 1857-1870] (ISSN 2320-5407). www.journalijar.com

Dr. Mukta Sharma
Prof. and Head, Department of Microbiology, Shree Bankey Bihari Dental College & Research Centre, Ghaziabad, (UP) India

DOI:

Article DOI: 10.21474/IJAR01/5252      
DOI URL: https://dx.doi.org/10.21474/IJAR01/5252

Download Full Paper

Download PDF No. of Downloads: 35 | No. of Views: 156