AIMS Microbiology, 2018, 4(4): 608-621. doi: 10.3934/microbiol.2018.4.608.

Research article

Export file:


  • RIS(for EndNote,Reference Manager,ProCite)
  • BibTex
  • Text


  • Citation Only
  • Citation and Abstract

New bacitracin-resistant nisin-producing strain of Lactococcus lactis and its physiological characterization

1 Laboratory of Biotechnology of Physiologically Active Compounds, Federal Research Centre “Fundamentals of Biotechnology”, Russian Academy of Sciences, Moscow 117312, Russia
2 Department of Molecular Biology, All-Russian Research Institute of Phytopathology, Bolshie Vyazemy, Moscow region 143050, Russia
3 FermLab LLC, Moscow 123592, Russia

Nisin A belonging to the class I bacteriocins and produced by Lactococcus lactis subsp. lactis is widely used in many countries as highly efficient and safe preservative preventing growth of undesirable bacteria in food products. Though this compound is efficient at very low concentrations, reduction of its manufacturing cost is still relevant problem. An increased nisin A production requires improved resistance of its producer to nisin. According to some studies, mechanisms of microbial resistance to nisin A and bacitracin have a similar basis, and the same transporters are used to export these antibiotics from cells. To obtain strains with improved growth rate and nisin A productivity, selection of spontaneous bacitracin-resistant L. lactis mutants followed by examination of their stability as well as physiological and fermentation characteristics was carried out. Spontaneous mutants were obtained by culturing of L. lactis VKPM B-2092 strain on selective bacitracin-containing agar medium. The obtained bacitracin-resistant strain FL-75 was characterized by accelerated growth rate, doubled biomass accumulation, and improved nisin A resistance. The nisin A productivity of FL-75 exceeded that of the parental strain by 25% reaching 8902 U/mL after 14-h cultivation. In addition, FL-75 was characterized by the improved resistance to oxidative stress that has never been reported earlier for bacitracin-resistant microorganisms. Based on the performed characterization of FL-75, we can consider it as a new independent strain promising for the industrial production of food and feed biopreservatives. Comparison of published data and the obtained results allowed us to suppose that the bacitracin resistance mutation in FL-75 is determined rather by an increased expression of a gene homologous to the bcrC gene of Bacillus sp. than by the activation of multidrug resistance mechanisms. The revealed resistance of FL-75 to bacitracin and oxidative stress can be regulated by a common transcription factor activating in response to various environmental stresses.
  Article Metrics

Keywords Lactococcus lactis; bacteriocins; nisin A; bacitracin; microbial drug resistance; oxidative stress

Citation: Vakhtang V. Dzhavakhiya, Elena V. Glagoleva, Veronika V. Savelyeva, Natalia V. Statsyuk, Maksim I. Kartashov, Tatiana M. Voinova, Alla V. Sergeeva. New bacitracin-resistant nisin-producing strain of Lactococcus lactis and its physiological characterization. AIMS Microbiology, 2018, 4(4): 608-621. doi: 10.3934/microbiol.2018.4.608


  • 1. Zacharof MP, Lovitt RW (2012) Bacteriocins produced by lactic acid bacteria: a review article. APCBEE Procedia 2: 50–56.    
  • 2. Hacker C, Christ NA, Duchardt-Ferner E, et al. (2015) The solution structure of the lantibiotic immunity protein NisI and its interactions with nisin. J Biol Chem 290: 28869–28886.
  • 3. Chatterjee C, Paul M, Xie L, et al. (2005) Biosynthesis and mode of action of lantibiotics. Chem Rev 105: 633–684.    
  • 4. Bierbaum G, Sahl HG (2009) Lantibiotics: mode of action, biosynthesis and bioengineering. Curr Pharm Biotechnol 10: 2–18.    
  • 5. Héchard Y, Sahl HG (2002) Mode of action of modified and unmodified bacteriocins from Gram-positive bacteria. Biochimie 84: 545–557.    
  • 6. Sun Z, Zhong J, Liang X, et al. (2009) Novel mechanism for nisin resistance via proteolytic degradation of nisin by the nisin resistance protein NSR. Antimicrob Agents Ch 53: 1964–1973.    
  • 7. Boziaris IS, Adams MR (1999) Effect of chelators and nisin produced in situ on inhibition and inactivation of Gram negatives. Int J Food Microbiol 53: 105–113.    
  • 8. Helander IM, Mattila-Sandholm T (2000) Permeability barrier of the Gram-negative bacterial outer membrane with special reference to nisin. Int J Food Microbiol 60: 153–161.    
  • 9. Draper LA, Ross RP, Hill C, et al. (2008) Lantibiotic immunity. Curr Protein Pept Sc 9: 39–49.    
  • 10. Draper LA, Cotter PD, Hill C, et al. (2015) Lantibiotic resistance. Microbiol Mol Biol Rev 79: 171–191.    
  • 11. Alkhatib Z, Abts A, Mavaro A, et al. (2012) Lantibiotics: how do producers become self-protected? J Biotechnol 159: 145–154.    
  • 12. Stein T, Heinzmann S, Solovieva I, et al. (2003) Function of Lactococcus lactis nisin immunity genes nisI and nisFEG after coordinated expression in the surrogate host Bacillus subtilis. J Biol Chem 278: 89–94.    
  • 13. AlKhatib Z, Lagedroste M, Fey I, et al. (2014) Lantibiotic immunity: inhibition of nisin mediated pore formation by NisI. PLoS One 9: e102246.    
  • 14. Kim WS, Hall RJ, Dunn NW (1997) The effect of nisin concentration and nutrient depletion on nisin production of Lactococcus lactis. Appl Microbiol Biotechnol 48: 449–453.    
  • 15. Martin-Visscher LA, Yoganathan S, Sit CS, et al. (2011) The activity of bacteriocins from Carnobacterium maltaromaticum UAL307 against Gram-negative bacteria in combination with EDTA treatment. FEMS Microbiol Lett 317: 152–159.    
  • 16. Balciunas EM, Martinez FAC, Todorov SD, et al. (2013) Novel biotechnological applications of bacteriocins: a review. Food Control 32: 134–142.    
  • 17. Gharsallaoui A, Oulahal N, Joly C, et al. (2016) Nisin as a food preservative. Part 1: physicochemical properties, antimicrobial activity, and main uses. Crit Rev Food Sci 56: 1262–1274.
  • 18. Dussault D, Vu KD, Lacroix M (2016) Enhancement of nisin production by Lactococcus lactis subsp. lactis. Probiotics Antimicro 8: 170–175.    
  • 19. Tafreshi SY, Mirdamadi S, Norouzian D, et al. (2010) Optimization of non-nutritional factors for a cost-effective enhancement of nisin production using orthogonal array method. Probiotics Antimicro 2: 267–273.    
  • 20. Kördikanlıoğlu B, Şimşek Ö, Saris PE (2015) Nisin production of Lactococcus lactis N8 with hemin-stimulated cell respiration in fed-batch fermentation system. Biotechnol Progr 31: 678–685.    
  • 21. González-Toledo SY, Domínguez-Domínguez J, García-Almendárez BE, et al. (2010) Optimization of nisin production by Lactococcus lactis UQ2 using supplemented whey as alternative culture medium. J Food Sci 75: 347–353.
  • 22. Field D, Cortter PD, Ross RP, et al. (2015) Bioengineering of the model lantibiotic nisin Bioengineered 6: 187–192.
  • 23. Simşek O, Con AH, Akkoç N, et al. (2009) Influence of growth conditions on the nisin production of bioengineered Lactococcus lactis strains. J Ind Microbiol Biotechnol 36: 481–490.    
  • 24. Cheigh CI, Park H, Choi HJ, et al. (2005) Enhanced nisin production by increasing genes involved in nisin Z biosynthesis in Lactococcus lactis subsp. lactis A164. Biotechnol Lett 27: 155–160.    
  • 25. Kumar PKR, Maschke HE, Friehs K, et al. Strategies for improving plasmid stability in genetically modified bacteria in bioreactors. Trends Biotechnol 9: 279–284.
  • 26. Arkin AP, Fletcher DA (2006) Fast, cheap and somewhat in control. Genome Biol 7: 114.    
  • 27. Jack BR, Leonard SP, Mishler DM, et al. (2015) Predicting the genetic stability of engineered DNA sequences with the EFM calculator. ACS Synth Biol 4: 939–943.    
  • 28. Podlesek Z, Herzog B, Comino A (1997) Amplification of bacitracin transporter genes in the bacitracin producing Bacillus licheniformis.FEMS Microbiol Lett 157: 201–205.    
  • 29. Hiron A, Falord M, Valle J, et al. (2011) Bacitracin and nisin resistance in Staphylococcus aureus: a novel pathway involving the BraS/BraR two-component system (SA2417/SA2418) and both the BraD/BraE and VraD/VraE ABC transporters. Mol Microbiol 81: 602–622.    
  • 30. Minareva LP, Bitteeva MB, Biryukov VV, et al. (2000) The development of the parameters of the nisin biosynthesis in flasks aimed at the selection of strains for the large-scale production. Biotekhnologiya 2: 29–36.
  • 31. Multon JL (1996) Quality control for foods and agricultural products, New York: Wiley-VCH.
  • 32. Popov AY, Isakova DM (1989) Biosynthesis of nisin by a butch fermentation of Streptococcus lactis. Biotekhnologiya 5: 583–587.
  • 33. Vasilyeva LA (2007) Statisticheskie metody v biologii, meditsine i sel'skom khozyaistve: uchebnoe posobie (Statistical methods in biology, medicine, and agriculture: a textbook), Novosibirsk: Institute of Cytology and Genetics.
  • 34. Podlesek Z, Comino A, Herzog B, et al. (2008) The role of the bacitracin ABC transporter in bacitracin resistance and collateral detergent sensitivity. FEMS Microbiol Lett 188: 103–106.
  • 35. Kitagawa N, Shiota S, Shibata Y, et al. (2011) Characterization of MbrC involved in bacitracin resistance in Streptococcusmutans. FEMS Microbiol Lett 318: 61–67.    
  • 36. Cain BD, Norton PJ, Eubanks W, et al. (1993) Amplification of the bacA gene confers bacitracin resistance to Escherichia coli. J Bacteriol 175: 3784–3789.    
  • 37. Ohki R, Tateno K, Okada Y, et al. (2003) A bacitracin-resistant Bacillus subtilis gene encodes a homologue of the membrane-spanning subunit of the Bacillus licheniformis ABC transporter. J Bacteriol 185: 51–59.    
  • 38. Lubelski J, Mazurkiewicz P, van Merkerk R, et al. (2004) YdaG and ydbA of Lactococcus lactis encode a heterodimeric ATP-binding cassette-type multidrug transporter. J Biol Chem 279: 34449–34455.    
  • 39. Kramer NE, van Hijum SAFT, Knol J, et al. (2006) Transcriptome analysis reveals mechanisms by which Lactococcus lactis acquires nisin resistance. Antimicrob Agents Ch 50: 1753–1761.    
  • 40. Navarre WW, Schneewind O (1999) Surface proteins of Gram-positive bacteria and mechanisms of their targeting to the cell wall envelope. Microbiol Mol Biol Rev 63: 174–229.
  • 41. Turner RJ, Aharonowitz Y, Weiner JH, et al. (2001) Glutatione is a target in tellurite toxicity and is protected by tellurite resistance determinants in Escherichia coli. J Microbiol 47: 33–40.
  • 42. Li Y, Molenaar D, Hugenholtz J, et al. (2003) Glutatione protects Lactococcus lactis against oxidative stress. Appl Environ Microb 69: 5739–5745.    
  • 43. Zhang J, Fu RY, Hugenholtz J, et al. (2007) Glutathione protects Lactococcus lactis against acid stress. Appl Environ Microb 73: 5268–5275.    
  • 44. Cretenet M, Gall GL, Wegmann U, et al. (2014) Early adaptation to oxygen is key to the industrially important traits of Lactococcus lactis ssp. cremoris during milk fermentation. BMC Genomics 15: 1054.


Reader Comments

your name: *   your email: *  

© 2018 the Author(s), licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution Licese (

Download full text in PDF

Export Citation

Copyright © AIMS Press All Rights Reserved