Synergistic Antimicrobial Effects of Nisin and ε-Poly-L-Lysine on Raw Beef During Cold Storage Against Major Foodborne Pathogens

Year 2025, Volume: 20 Issue: 2, 94 - 103, 27.08.2025

Mehmet Emin Aydemir , Teyfik Çelen , Serap Kılıç Altun , Mehmet Nuri Giraz , Emine Betül Tuğbay , Ayşegül Keşküş

https://doi.org/10.17094/vetsci.1691935

Abstract

With increasing consumer demand for natural food preservatives, the use of antimicrobial substances such as nisin and epsilon-poly-L-lysine (ε-PL) in meat products has gained attention. This study aimed to evaluate the synergistic effects of nisin and ε-PL on the viability of key foodborne pathogens—Escherichia coli O157:H7, Salmonella Typhimurium (Gram-negative), and Listeria monocytogenes (Gram-positive)—in raw red meat. In addition, the study evaluated the impact of treatments on physicochemical characteristics and color stability. Two combinations of nisin and ε-PL were tested: Mix 1 (400 IU/g nisin + 20 μg/g ε-PL) and Mix 2 (800 IU/g nisin + 40 μg/g ε-PL), applied to raw beef samples inoculated with the pathogens. Samples were stored at 4 °C for 16 days, and microbiological (pathogen counts, total mesophilic aerobic bacteria, mold, and yeast), physicochemical (pH, water-holding capacity), and color (L*, a*, b*) analyses were performed at intervals (days 0, 4, 8, 12, and 16). Mix 2 showed the most potent antimicrobial activity, decreasing L. monocytogenes counts to 2.15 log CFU/g by day 8. Significant reductions were also recorded for E. coli O157:H7 and Salmonella Typhimurium. Additionally, the mixtures suppressed pH increases, maintained color stability, and improved water retention. In conclusion, the free-form combinations of nisin and ε-PL effectively inhibited microbial growth, preserved meat quality, and extended shelf life, highlighting their potential as natural preservatives in the meat industry. This study provides novel evidence on the synergistic use of free-form nisin and ε-PL in raw beef, offering a valuable contribution to the development of natural preservation methods in meat products.

Keywords

Nisin , epsilon-poly-L-lysine (ε-PL) , meat , food biopreservation , natural antimicrobials

Nisin ve ε-Poli-L-Lizin'in Soğuk Depolama Sırasında Çiğ Sığır Etinde Başlıca Gıda Kaynaklı Patojenlere Karşı Sinerjik Antimikrobiyal Etkileri

Abstract

Doğal gıda koruyucularına yönelik artan tüketici talebiyle birlikte, et ürünlerinde nisin ve epsilon-poly-L-lysine (ε-PL) gibi antimikrobiyal maddelerin kullanımı dikkat çekmiştir. Bu çalışma, nisin ve ε-PL'nin çiğ kırmızı ette gıda kaynaklı önemli patojenlerin Escherichia coli O157:H7, Salmonella Typhimurium (Gram-negatif) ve Listeria monocytogenes (Gram-pozitif) canlılığı üzerindeki sinerjik etkilerini değerlendirmeyi amaçlamıştır. Çalışmada ayrıca uygulamaların fizikokimyasal özellikler ve renk stabilitesi üzerindeki etkisi de değerlendirilmiştir. Nisin ve ε-PL'nin iki kombinasyonu test edilmiştir: Karışım 1 (400 IU/g nisin + 20 μg/g ε-PL) ve Karışım 2 (800 IU/g nisin + 40 μg/g ε-PL), patojenlerle inoküle edilmiş çiğ sığır eti örneklerine uygulanmıştır. Örnekler 16 gün boyunca 4 °C'de saklanmış ve aralıklarla (0, 4, 8, 12 ve 16. günler) mikrobiyolojik (patojen sayıları, toplam mezofilik aerobik bakteri, küf ve maya), fizikokimyasal (pH, su tutma kapasitesi) ve renk (L*, a*, b*) analizleri yapılmıştır. Karışım 2, L. monocytogenes sayısını 8. günde 2,15 log CFU/g'a düşürerek en güçlü antimikrobiyal aktiviteyi göstermiştir. E. coli O157:H7 ve S. Typhimurium için de önemli azalmalar kaydedilmiştir. Ayrıca, karışımlar pH artışlarını bastırmış, renk stabilitesini korumuş ve su tutma özelliğini geliştirmiştir. Sonuç olarak, nisin ve ε-PL'nin serbest form kombinasyonları mikrobiyal büyümeyi etkili bir şekilde engellemiş, et kalitesini korumuş ve raf ömrünü uzatmıştır.

Keywords

Nisin , epsilon-poli-L-lisin (ε-PL) , et , gıda biyokoruyucu , doğal antimikrobiyaller

References

• 1. Tan L, Ni Y, Xie Y, et al. Next generation meat preservation: integrating nano-natural ingredients to overcome barriers and opportunities. Crit Rev Food Sci Nutr. 2024;64(33):12720–12743.
• 2. Das AK, Nanda PK, Das A, Biswas S. Hazards and safety issues of meat and meat products. In: In Food safety and human health. Academic Press. 2019;145-168.
• 3. Acaröz U, Acaröz DA. Microbiology of meat and meat products. Turkiye Klinikleri Food Sci Spec Top. 2021;7(2):75-78.
• 4. Shaltout FA, Barr AAH, Abdelaziz ME. Pathogenic microorganisms in meat products. Biomed Sci Tech Res J. 2022;41(4):32836-32843.
• 5. Gutema FD, Agga GE, Abdi RD, et al. Assessment of hygienic practices in cattle abattoirs and retail stores in Bishoftu, Ethiopia: implications for public health. Int J Environ Res Public Health. 2021;18(5):2729.
• 6. Incili GK, Aydemi̇r ME, Akgöl M, et al. Effect of rheum ribes L. juice on the survival of Listeria monocytogenes, Escherichia coli O157: H7 and Salmonella Typhimurium and chemical quality on vacuum packaged raw beef. LWT. 2021;150:112016.
• 7. Özel B, Şimşek Ö, Akçelik M, Saris PEJ. Innovative approaches in nisin production. Appl Microbiol Biotechnol. 2018;102:6299-6307.
• 8. Gharsallaoui A, Oulahal N, Joly C, Degraeve P. Nisin as a food preservative: Part 1: Physicochemical properties, antimicrobial activity and main uses. Crit Rev Food Sci Nutr. 2015;56(8):1262-1274.
• 9. Zinoviadou KG, Koutsoumanis KP, Biliaderis CG. Physical and thermo-mechanical properties of whey protein isolate films containing antimicrobials, and their effect against spoilage flora of fresh beef. Food Hydrocoll. 2010;24(1):49–59.
• 10. Hyldgaard M, Mygind T, Vad BS, Stenvang M, Otzen DE, Meyer RL. The antimicrobial mechanism of action of epsilon-poly-L-lysine. Appl Environ Microbiol. 2014;80(24):7758–7770.
• 11. Bortolotto FCK, Farfan MHDR, Jede NCK, Danielski GM, Macedo REF. Nisin and ε-poly-L-lysine as natural antimicrobials towards spoilage-associated Lactobacillus plantarum. Ciênc Rural (Online). 2021;e20200423-e20200423.
• 12. Najjar MB, Kashtanov D, Chikindas ML. ε-Poly-L-lysine and nisin A act synergistically against Gram-positive food-borne pathogens Bacillus cereus and Listeria monocytogenes. Lett Appl Microbiol. 2007;45(1):13–18.
• 13. Liu F, Liu M, Du L, et al. Synergistic antibacterial effect of the combination of ɛ-polylysine and nisin against Enterococcus faecalis. J Food Prot. 2015;78(12):2200–2206.
• 14. FDA F. Guidance for industry: bioanalytical method validation. Accessed Sep 5, 2023. http://www.fda.gov/cder/Guidance/4252fnl.pdf.
• 15. ISO 21527-1. Microbiology of food and animal feeding stuffs: Horizontal method for the enumeration of yeasts and moulds. 2008.
• 16. Aydemir ME, Sezer E, Giraz MN. Effect of pine cone vinegar on the survival of Listeria monocytogenes and Salmonella typhimurium and some physico-chemical properties in raw beef. Kocatepe Vet J. 2024;17(4):299–307.
• 17. Aydemir ME, Altun SK, Takım K, Yilmaz MA, Yalçin H. Inhibitory effect of homemade hawthorn vinegar-based marinade on Nε-(carboxymethyl) lysine and Nε-(carboxyethyl) lysine formation in beef tenderloins. Meat Sci. 2024;214:109535.
• 18.Zimet P, Mombrú ÁW, Faccio R, et al. Optimization and characterization of nisin-loaded alginate-chitosan nanoparticles with antimicrobial activity in lean beef. LWT. 2018;91:107–116.
• 19. Khan I, Oh DH. Integration of nisin into nanoparticles for application in foods. Innov Food Sci Emerg Technol. 2016;34:376–384.
• 20. Mansur AR, Song EJ, Cho YS, et al. Comparative evaluation of spoilage-related bacterial diversity and metabolite profiles in chilled beef stored under air and vacuum packaging. Food Microbiol. 2019;77:166–172.
• 21.Ibrahim SA, Ayivi RD, Zimmerman T, et al. Lactic acid bacteria as antimicrobial agents: Food safety and microbial food spoilage prevention. Foods. 2021;10(12):3131.
• 22. Geornaras I, Yoon Y, Belk KE, Smith GC, Sofos JN. Antimicrobial activity of ɛ-polylysine against Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes in various food extracts. J Food Sci. 2007;72(8):M330–334.
• 23. Kang J, Stasiewicz MJ, Murray D, Boor KJ, Wiedmann M, Bergholz TM. Optimization of combinations of bactericidal and bacteriostatic treatments to control Listeria monocytogenes on cold-smoked salmon. Int J Food Microbiol. 2014;179:1–9.
• 24. Alirezalu K, Yaghoubi M, Poorsharif L, et al. Antimicrobial polyamide-alginate casing incorporated with nisin and ε-polylysine nanoparticles combined with plant extract for inactivation of selected bacteria in nitrite-free frankfurter-type sausage. Foods. 2021;10(5):1003.
• 25. Lee HJ, Heo Y, Kim HJ, et al. Bactericidal effect of combination of atmospheric pressure plasma and nisin on meat products inoculated with Escherichia coli O157:H7. Food Sci Anim Resour. 2023;43(3):402.
• 26. Yaghoubi M, Alirezalu K, Mazloomi SM, et al. Enhancing beef sausage packaging with calcium alginate active film infused with nisin and ε-polylysine nanoparticles and beetroot extract. LWT. 2024;191:115665.
• 27. Lee DU, Park YJ, Yu HH, et al. Antimicrobial and antibiofilm effect of ε-polylysine against Salmonella enteritidis, Listeria monocytogenes, and Escherichia coli in tryptic soy broth and chicken juice. Food. 2021;10(9):2211.
• 28. Tan Z, Bo T, Guo F, Cui J, Jia S. Effects of ε-poly-l-lysine on the cell wall of Saccharomyces cerevisiae and its related antimicrobial mechanism. Int J Biol Macromol. 2018;118:2230–2236.
• 29. Ayaseh A, Alirezalu K, Yaghoubi M, et al. Production of nitrite-free frankfurter-type sausages by combining ε-polylysine with beetroot extracts: An assessment of microbial, physicochemical, and sensory properties. Food Biosci. 2022;49:101936.
• 30. Bhat AA, Ahmed A, Dar MA, Achir A, Pagrut N. Effect of different levels of nisin on the microbial quality of chicken cutlets. J Livestock Sci. 2015;6:47–51.
• 31. Sayadi M, Mojaddar Langroodi A, Amiri S, Radi M. Effect of nanocomposite alginate‐based film incorporated with cumin essential oil and TiO₂ nanoparticles on chemical, microbial, and sensory properties of fresh meat/beef. Food Sci Nutr. 2022;10(5):1401–1413.
• 32. Bangar SP, Sharma N, Bhardwaj A, Phimolsiripol Y. Lactic acid bacteria: A bio-green preservative against mycotoxins for food safety and shelf-life extension. Qual Assur Saf Crops Foods. 2022;14(2):13–31.
• 33. Bağdatli A, Kayaardi S. Influence of storage period and packaging methods on quality attributes of fresh beef steaks. CyTA J Food. 2015;13(1):124–133.
• 34. Wyrwisz J, Półtorak ANDRZEJ, Zalewska M, Zaremba R, Wierzbicka A. Analysis of relationship between basic composition, pH, and physical properties of selected bovine muscles. Bull Vet Inst Pulawy. 2012;56(3):403–409.
• 35. El Adab S, Hassouna M. Proteolysis, lipolysis and sensory characteristics of a Tunisian dry fermented poultry meat sausage with oregano and thyme essential oils. J Food Saf. 2016;36(1):19–32