Volume : 10, Issue : 7, JUL 2024

PRODUCTION, EXTRACTION AND PURIFICATION OF BACTERIOCIN FROM LACTIC ACID BACTERIA AND EVALUATING ITS ANTIBACTERIAL POTENTIAL AGAINST FOOD SPOILAGE BACTERIA

SHEHMA SHUKKOOR, RAJESWARI, K

Abstract

Preservation of foods during storage is considered important step in seafood industries. Hence the aim of this research is to analyze the effect of bacteriocin as bio-preservative agents against the seafood pathogens. Biopreservation using bacteriocin from two lactic acid bacteria, Lactobacillus acidophilus – L1 and Lactobacillus sporogenes – L2 against different food pathogens were studied in the present research. Bacteriocin from Lactic acid bacteria were purified using dialysis method; and SDS-PAGE analysis was carried out to identify the protein bands of bacteriocin. Antibacterial activity of bacteriocin concentrations was evaluated using standard agar well diffusion method. SDS-PAGE results of the purified fractions of L1 and L2 showed three distinct protein bands with molecular weight of 55kDa, 30kDa, 10kDa; and five distinct bands with molecular weight of 90kDa, 76kDa, 54kDa, 24kDa and 20kDa respectively. Antibacterial activity test results of L1 bacteriocin revealed inhibitory zones ranging from 13.9mm to 19.3mm against all test bacterial cultures. And L2 bacteriocin exhibited inhibitory zones ranging from 14.9mm to 20.3mm against all test bacteria. The inhibitory effect of bacteriocins on the viable cells of food pathogens showed maximum cell inhibition. The bacteriocin exposed to Salmonella cells reduced maximum of about 90% followed by Staphylococcus aureus upto 86%. As future studies, the antibacterial action of the extracted nutraceutical compounds shall be increased by handling further purification techniques like chromatography and other methods. The hurdle factors need to be studied in detail as future study to identify the best combination factors for food preservation against food spoilage microorganisms.

Article : Download PDF

Cite This Article

-

Article No : 2

Number of Downloads : 106

References

1. Abril GA, Villa GT, Barros-Velazquez J, Canas B, Sanchez-Perez A, Calo-Mata P. (2020). Staphylococcus aureus Exotoxins and Their Detection in the Dairy Industry and Mastitis. Toxins. 2(9):537-541.

2. Akbar Ali and Anal AK. (2014). Occurrence of Staphylococcus aureus and evaluation of antistaphylococcal activity of Lactococcus lactis subsp. lactis in ready-to-eat poultry meat. Annals of Microbiology. 64(1):131-138.

3. Alamri MS, Akram AA, Abdellatif AM, Shahzad Hussain, Mohamed AI, Ghalia Shamlan. (2021). Food packaging’s materials: A food safety perspective. (2021). Saudi Journal of Biological Sciences. 28(8):4490-4499.

4. Alomari A, Matter IR, Almola AH. (2022). An Overview of Bacteriocins. Samarra. Journal of Pure Applied Science. 4(2):58-72.

5. Amit SK, Uddin MM, Rahman R. (2017). A review on mechanisms and commercial aspects of food preservation and processing. (2017). Agriculture Food Security.6:51-58.

6. Cintas LM, Herranz C, Hernandez PE, Casaus MP and Nes LF. Review: Bacteriocins of lactic acid bacteria. (2001). Food Science and Technology International. 7:281-305.

7. Cleveland J, Montville TJ, Nes IF and Chikindas ML. (2001).  Bacteriocins: safe, natural antimicrobials for food preservation. International Journal of Food Microbiology. 71:1-20.

8. Cleveland J, Montville TJ, Nes IF and Chikindas ML. (2021). Bacteriocins: safe, natural antimicrobials for food preservation. International Journal of Food Microbiology. 7:1-20.

9. Cotter, P. D., Hill, C., & Ross, R. P. (2005). Bacteriocins: developing innate immunity for food.Nature Reviews Microbiology, 3(10):777-788.

10. Cotter, P. D., Ross, R. P., & Hill, C. (2013). Bacteriocins—a viable alternative to antibiotics?. Nature Reviews Microbiology, 11(2):95-105.

11. Galvez A, Abriouel H, Lopez RL and Omar NB. (2007). Bacteriocin-based strategies for food biopreservation, International Journal of Food Microbiology. 120:51-70.

12. Goraya MU, Ashraf M., Rahman S. U. and Habib A. (2013). Determination of antibacterial activity of bacteriocins of lactic acid producing bacteria. Journal of Molecular Biology. 1(1):8-10.

13. Hassan MU, Nayab H, Rehman TU, Williamson MP, Haq KU, Shafi N. (2020). Characterisation of Bacteriocins Produced by Lactobacillus spp. Isolated from the Traditional Pakistani Yoghurt and Their Antimicrobial Activity against Common Foodborne Pathogens. Biomedical Research International. 828-830.

14. Hoa ND, Wouters R, Wille M, Thanh V, Dong TK, Hao NV. (2009). A fresh food maturation diet with an adequate HUFA composition for brood-stock nutrition studies in black tiger shrimp Penaeus monodon. Aquaculture. 297:116-121.

15. Islam, R., Hossain, Alam, K., Uddin, Rony, M.H., Imran, A.S. and Alam, F. (2020) Antibacterial Activity of Lactic Acid Bacteria and Extraction of Bacteriocin Protein. Advances in Bioscience and Biotechnology, 11, 49-59.

16. Jack RW, Tagg JR and Ray B (1995). Bacteriocins of Gram-positive bacteria. Microbiological Review. 59:171-200.

17. Khan H, Flint S and Yu PL. Enterocins in food preservation. (2010). International Journal of Food Microbiology. 141:1- 10.

18. Klein, A., Wojdyla, J. A., Joshi, A., Josts, I., McCaughey, L. C., Housden, N. G., & Kleanthous, C. (2016). Structural and biophysical analysis of nuclease protein antibiotics. Biochemical Journal. 473(18):2799-2812

19. Kyule DN, Maingi JM, Njeru EM, Nyamache AK. (2022). Molecular Characterization and Diversity of Bacteria Isolated from Fish and Fish Products Retailed in Kenyan Markets. International Journal of Food Science. 1-12.

20. Leroi F, Cornet J, Chevalier F, Cardinal M, Coeuret G, Chaillou S. (2015). Selection of bioprotective cultures for preventing cold-smoked salmon spoilage. International Journal of Food Microbiology. 213:79-87.

21. Montville TJ and Chen Y. (1998). Mechanistic Action of Pediocin and Nisin: Recent Progress and Unresolved Questions. Applied Microbiology and Biotechnology. 50:267-274.

22. Motta AS and Brandelli A. (2008). Evaluation of environmental conditions for production of bacteriocin like substance by Bacillus sp. strain P34. World Journal of Microbiology and Biotechnology. 24: 641-646.

23. Muhammad Zahid, Muhammad Ashraf, Muhammad Arshad, Ghulam Muhammad, Aqeela Yasmin and Hafiz Muhammad. (2015). Antimicrobial Activity of Bacteriocins isolated from Lactic Acid Bacteria against Resistant Pathogenic Strains. International Journal of Nutrition and Food Science. 4(3):326-331.

24. Nawrocki KL, Crispell EK and McBride SM. (2014). Antimicrobial peptide resistance mechanisms of Gram-positive bacteria. Antibiotics. 3:461-492.

25. Negash, A. W., & Tsehai, B. A. (2020). Current applications of bacteriocin. International Journal of Microbiology. 437-489.

26. Nivedita and Neha. (2008). Antibacterial activitiy and characterization of bacteriocin of Bacillus mycoides isolated from whey. Indian Journal of Biotechnology. 7:117-121.

27. Pravin Deshmukh1 and Prakash Thorat. (2018). Extraction and Purification of Bacteriocin from Lactobacillus brevis CB-2 and Lactobacillus zymae WHL-7 for their antimicrobial activity. DAV International Journal of Science. 7(1):1-8.

28. Preciado, G. M., Michel, M. M., Villarreal-Morales, S. L., Flores-Gallegos, A. C., Aguirre-Joya,   J.,   Morlett-Chávez,  J.,  ...  &  Rodríguez-Herrera,   R.   (2016). Bacteriocins and its use for multidrug-resistant bacteria control. Antibiotic Resistance. 329-349.

29. Prince, A., Sandhu, P., Ror, P., Dash, E., Sharma, S., Arakha, M., Saleem, M. (2016).  Lipid-II independent antimicrobial mechanism of nisin depends on its crowding    and    degree    of oligomerization. Scientific Reports. 6(1):1-15.

30. Qiao, Z., Chen, J., Zhou, Q., Wang, X., Shan, Y., Yi, Y., Liu, B., Zhou, Y., & Lü, X. (2021). Purification, characterization, and mode of action of a novel bacteriocin BM173 from Lactobacillus crustorum MN047 and its effect on biofilm formation of Escherichia coli and Staphylococcus aureus. Journal of Dairy Science. 104(2):1474-1483

31. Rahmeh, R., Akbar, A., Alonaizi, T., Kishk, M., Shajan, A., & Akbar, B. (2020). Characterization and application of antimicrobials produced by Enterococcus faecium S6 isolated from raw camel milk. Journal of Dairy Science. 103(12):106-115.

32. Sahar K, Mohammad R, Hosna H, Mahmoud B, Hassan H,  Mahmoodnia L. (2017). Isolation and identification of probiotic Lactobacillus from local dairy and evaluating their antagonistic effect on pathogens. International Journal of Pharmaceutical Investigation. 7(3):137–141.

33. Saini RV, Vaid P, Saini NK, Siwal SS, Gupta VK, Thakur VK. (2021). Recent advancements in the technologies detecting food spoiling agents. Journal of Functional Biomaterials. 12(4):67-72.

34. Sivaramasamy Elayaraja, Neelamegam Annamalai, Packiyam Mayavu, Thangavel Balasubramanian. (2014). Production, purification and characterization of bacteriocin from Lactobacillus murinus AU06 and its broad antibacterial spectrum. Asian Pacific Journal of Tropical Biomedicine. 4(1):305-311

35. Teneva D and Denev P. (2023). Biologically Active Compounds from Probiotic Microorganisms and Plant Extracts Used as Biopreservatives. Microorganisms. 11(8):1896.

36. Tolpekina T.V., den Otter W.K., Briels W.J. (2004). Nucleation Free Energy of Pore Formation in an Amphiphilic Bilayer Studied by Molecular Dynamics Simulations. Journal of Chemistry and Physics. 121:60–66.

37. Van Belkum M.J., Kok J., Venema G., Holo H., Nes I.F., Konings W.N., Abee T. (1991). The Bacteriocin Lactococcin a Specifically Increases Permeability of Lactococcal Cytoplasmic Membranes in a Voltage-Independent, Protein-Mediated Manner. Journal of Bacteriology. 173:7934–7941

38. Walker,  D.,  Mosbahi,  K.,  Vankemmelbeke,  M.,  James,  R.,  &  Kleanthous,  C. (2007).  The  role  of  electrostatics  in  colicin  nuclease  domain  translocation  into bacterial cells. Journal of Biological Chemistry. 282(43):31389-31397.

39. Weijia Li, Zhou and Yuyin Xu. Study of the in vitro cytotoxicity testing of medical devices. (2015). Biomedical Reports. 3(5):617–620.