Controlled release of nisin from Neusilin particles to enhance food safety of sour curd cheese

  • Maik Szendy
  • Florian Westhaeuser
  • Barbara Baude
  • Jessica Reim
  • Lars Dähne
  • Matthias NollEmail author
Original Article


Nisin is frequently added as food additive to soft cheese to increase food safety against foodborne pathogens like Listeria monocytogenes. The goal of this study was the extension of the antimicrobial activity of nisin in sour curd cheese (SCC) by self-releasing adsorbed nisin from Neusilin UFL2 over production-based pH shift. First, the antimicrobial activity of nisin adsorbed to Neusilin UFL2 (UFL2-N) and free nisin was investigated in BHI broth at a pH range from 7.5 to 4.5 for each of six L. monocytogenes field isolates. UFL2-N showed similar minimal inhibition concentration to L. monocytogenes over time as free nisin. Distribution of nebulized, fluorescence-labelled UFL2 was homogenous on SCC surface. Thereafter, SCC surface was inoculated with L. monocytogenes and 0.004, 0.013, 0.026, and 0.132 mg mL−1 UFL2-N or free nisin. In SCC, L. monocytogenes was below quantification limit at 0.132 mg mL−1 UFL2-N or free nisin after 2 days of ripening. Collectively, UFL2-N enabled a slow release and antilisterial activity in vitro as well as in cheese manufacturing.


Adsorption Food safety Listeria monocytogenes Neusilin UFL2 Nisin 



This research was financially supported by Federal Ministry for Economic Affairs and Energy on the basis of a decision by the German Bundestag (KF3083302SK3) and technical alliance Oberfranken (TAO).

Supplementary material

13197_2019_3577_MOESM1_ESM.pptx (710 kb)
Supplementary material 1 (PPTX 710 kb)


  1. Aasen IM, Markussen S, Møretrø T, Katla T, Axelsson L, Naterstad K (2003) Interactions of the bacteriocins sakacin P and nisin with food constituents. Int J Food Microbiol 87:35–43. CrossRefGoogle Scholar
  2. Allerberger F (2003) Listeria: growth, phenotypic differentiation and molecular microbiology. FEMS Immunol Med Microbiol 3:183–189CrossRefGoogle Scholar
  3. Belitz H-D, Grosch W, Schieberle P (2001) Milch und Milchprodukte. In: Belitz H-D, Grosch W, Schieberle P (eds) Lehrbuch der Lebensmittelchemie. Springer, Heidelberg, pp 490–535CrossRefGoogle Scholar
  4. Bhatti M, Veeramachaneni A, Shelef LA (2004) Factors affecting the antilisterial effects of nisin in milk. Int J Food Microbiol 97:215–219CrossRefGoogle Scholar
  5. Buettner C, Noll M (2018) Differences in microbial key players in anaerobic degradation between biogas and sewage treatment plants. Int Biodeterior Biodegrad 133:124–132. CrossRefGoogle Scholar
  6. Cheroutre-Vialette M, Lebert I, Hebraud M, Labadie J, Lebert A (1998) Effects of pH or aw stress on growth of Listeria monocytogenes. Int J Food Microbiol 42:71–77. CrossRefGoogle Scholar
  7. Chollet E, Sebti I, Martial-Gros A, Degraeve P (2008) Nisin preliminary study as a potential preservative for sliced ripened cheese: NaCl, fat and enzymes influence on nisin concentration and its antimicrobial activity. Food Control 19:982–989. CrossRefGoogle Scholar
  8. De Martinis ECP, Crandall AD, Mazzotta AS, Montville TJ (1997) Influence of pH, salt, and temperature on nisin resistance in Listeria monocytogenes. J Food Prot 60:420–423CrossRefGoogle Scholar
  9. Delves-Broughton J, Blackburn P, Evans RJ, Hugenholtz J (1996) Applications of the bacteriocin, nisin. Antonie Van Leeuwenhoek 69:193–202. CrossRefGoogle Scholar
  10. EU (2011) Commission Regulation (EU) No 1129/2011 of 11 November 2011 amending Annex II to Regulation (EC) No 1333/2008 of the European Parliament and of the Council by establishing a union list of food additives. Commission Regulation (EU) No 1129/2011 of 11 November 2011 amending Annex II to Regulation (EC) No 1333/2008 of the European Parliament and of the Council by establishing a union list of food additivesGoogle Scholar
  11. Ferreira MA, Lund BM (1996) The effect of nisin on Listeria monocytogenes in culture medium and long-life cottage cheese. Lett Appl Microbiol 22:433–438CrossRefGoogle Scholar
  12. Fretz R, Sagel U, Ruppitsch W, Pietzka AT, Stöger A, Huhulescu S, Heuberger S, Pichler J, Much P, Pfaff G, Stark K, Prager R, Flieger A, Feenstra O, Allerberger F (2010) Listeriosis outbreak caused by acid curd cheese ‘Quargel’, Austria and Germany 2009. Euro Surveill. Google Scholar
  13. Gill C, Reichel M (1989) Growth of the cold-tolerant pathogens Yersinia enterocolitica, Aeromonas hydrophila and Listeria monocytogenes on high-pH beef packaged under vacuum or carbon dioxide. Food Microbiol 6:223–230CrossRefGoogle Scholar
  14. 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 Microbiol. Google Scholar
  15. Ibarguren C, Naranjo PM, Stötzel C, Audisio MC, Sham EL, Farfán Torres EM, Müller FA (2014) Adsorption of nisin on raw montmorillonite. Appl Clay Sci 90:88–95. CrossRefGoogle Scholar
  16. Kudamala S, Murthy KVR (2017) Applicability of a novel carrier, Neusilin UFL2, for the preparation of domperidone liquisolid tablets. World J Pharm Pharam Sci 6:1662–1671Google Scholar
  17. Ma Y-H, Wang Q, Gong J, Wu XY (2016) Formulation of granules for site-specific delivery of an antimicrobial essential oil to the animal intestinal tract. J Pharm Sci 105:1124–1133. CrossRefGoogle Scholar
  18. Mallappa MK, Kesarla R, Banakar S (2015) Calcium alginate-Neusilin US2 nanocomposite microbeads for oral sustained drug delivery of poor water soluble drug aceclofenac sodium. J Drug Deliv. Google Scholar
  19. Martinez-Rios V, Dalgaard P (2018) Prevalence of Listeria monocytogenes in European cheeses: a systematic review and meta-analysis. Food Control 84:205–214. CrossRefGoogle Scholar
  20. McElhatton A, Marshall RJ (2007) Food safety: a practical and case study approach. Springer, New YorkCrossRefGoogle Scholar
  21. Meira SMM, Jardim AI, Brandelli A (2015) Adsorption of nisin and pediocin on nanoclays. Food Chem 188:161–169. CrossRefGoogle Scholar
  22. Ming X, Daeschel MA (1993) Nisin resistance of foodborne bacteria and the specific resistance responses of Listeria monocytogenes Scott A. J Food Prot 56:944–948CrossRefGoogle Scholar
  23. Muyzer G, De Waal E, Uitterlinden A (1993) Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Appl Environ Microbiol 59:695–700Google Scholar
  24. Nogva HK, Naterstad KR, Holck A, Lillehaug D (2000) Application of 5′-nuclease PCR for quantitative detection of Listeria monocytogenes in pure cultures, water, skim milk, and unpasteurized whole milk. Appl Environ Microbiol 66:4266–4271CrossRefGoogle Scholar
  25. Noll M, Kleta S, Al Dahouk S (2018) Antibiotic susceptibility of 259 Listeria monocytogenes strains isolated from food, food-processing plants and human samples in Germany. J Infect Public Health 11:572–577. CrossRefGoogle Scholar
  26. Pawar DD, Malik S, Bhilegaonkar KN, Barbuddhe SB (2000) Effect of nisin and its combination with sodium chloride on the survival of Listeria monocytogenes added to raw buffalo meat mince. Meat Sci 56:215–219. CrossRefGoogle Scholar
  27. Peyratout CS, Dähne L (2004) Tailor-made polyelectrolyte microcapsules: from multilayers to smart containers. Angew Chem Int Ed Engl 43:3762–3783. CrossRefGoogle Scholar
  28. Prombutara P, Kulwatthanasal Y, Supaka N, Sramala I, Chareonpornwattana S (2012) Production of nisin-loaded solid lipid nanoparticles for sustained antimicrobial activity. Food Control 24:184–190. CrossRefGoogle Scholar
  29. Ramsaran H, Chen J, Brunke B, Hill A, Griffiths MW (1998) Survivial of bioluminescent Listeria monocytogenes and Escherichia coli 0157:H7 in soft cheeses. J Dairy Sci 81:1810–1817. CrossRefGoogle Scholar
  30. Rogga KJ, Samelis J, Kakouri A, Katsiari MC, Savvaidis IN, Kontominas MG (2005) Survival of Listeria monocytogenes in Galotyri, a traditional Greek soft acid-curd cheese, stored aerobically at 4°C and 12°C. Int Dairy J 15:59–67. CrossRefGoogle Scholar
  31. Santaniello M, Giannini G (2016) A versatile and stable mixture of fish oil and resveratrol in a powder formulation. Nutr Food Technol Google Scholar
  32. Sulzer G, Busse M (1991) Growth inhibition of Listeria spp. on camembert cheese by bacteria producing inhibitory substances. Int J Food Microbiol 14:287–296. CrossRefGoogle Scholar
  33. Sun Z, Zhong J, Liang X, Liu J, Chen X, Huan L (2009) Novel mechanism for nisin resistance via proteolytic degradation of nisin by the nisin resistance protein NSR. Antimicrob Agents Chemother 53:1964–1973. CrossRefGoogle Scholar
  34. Were LM, Bruce B, Davidson PM, Weiss J (2004) Encapsulation of nisin and lysozyme in liposomes enhances efficacy against Listeria monocytogenes. J Food Prot 67:922–927. CrossRefGoogle Scholar
  35. Yen LC, Sofos JN, Schmidt GR (1991) Effect of meat curing ingredients on thermal destruction of Listeria monocytogenes in ground pork. J Food Prot 54:408–412. CrossRefGoogle Scholar

Copyright information

© Association of Food Scientists & Technologists (India) 2019

Authors and Affiliations

  1. 1.Institute for Bioanalysis, Department of Applied SciencesCoburg University of Applied Sciences and ArtsCoburgGermany
  2. 2.Surflay Nanotec GmbHBerlinGermany

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