Antonie van Leeuwenhoek

, Volume 112, Issue 2, pp 297–304 | Cite as

Novel antimicrobial peptides produced by Candida intermedia LAMAP1790 active against the wine-spoilage yeast Brettanomyces bruxellensis

  • Rubén Peña
  • María Angélica GangaEmail author
Original Paper


Brettanomyces bruxellensis negatively impacts on the sensorial quality of wine by producing phenolic compounds associated with unpleasant odors. Thus, the control of this spoilage yeast is a critical factor during the winemaking process. A recent approach used to biocontrol undesired microorganisms is the use of yeast released antimicrobial peptides (AMPs), but this strategy has been poorly applied to wine-related microorganisms. The aim of this study was to evaluate the antifungal capacity of Candida intermedia LAMAP1790 against wine-spoilage strains of B. bruxellensis and fermentative strains of Saccharomyces cerevisiae, and also to determine the chemical nature of the compound. The exposure of strains to the supernatant of C. intermedia saturated cultures showed antifungal activity against B. bruxellensis, without affecting the growth of S. cerevisiae. By fractionation and concentration of C. intermedia supernatants, it was determined that the antifungal activity was related to the presence of heat-labile peptides with molecular masses under 5 kDa. To our knowledge, this is the first report of AMPs secreted by C. intermedia that control B. bruxellensis. This could lead to the development of new biocontrol strategies against this wine-spoilage yeast.


Antimicrobial peptides Antifungal activity Biocontrol Brettanomyces bruxellensis Candida intermedia Wine-spoilage yeasts 



This work has been supported by Proyecto Fortalecimiento USACH USA 1398_GM181622 Grant. Rubén Peña is funded by the Comisión Nacional de Investigación Científica y Tecnológica CONICYT-PCHA/Doctorado Nacional/2013-21130439 Doctoral Fellowship.

Author’s contributions

MAG conceived and designed the study. RP performed research and analyzed data. MAG and RP wrote the paper.

Conflict of interest

The authors hereby declare they do not have any conflict of interest associated to this work.


  1. Acuña-Fontecilla A, Silva-Moreno E, Ganga MA, Godoy L (2017) Evaluation of antimicrobial activity from native wine yeast against food industry pathogenic microorganisms. CYTA J Food 15(3):457–465CrossRefGoogle Scholar
  2. Albergaria H, Francisco D, Gori K, Arneborg N, Gírio F (2010) Saccharomyces cerevisiae CCMI 885 secretes peptides that inhibit the growth of some non-Saccharomyces wine-related strains. Appl Microbiol Biotechnol 86(3):965–972CrossRefGoogle Scholar
  3. Avramova M, Cibrario A, Peltier E, Coton M, Coton E et al (2018) Brettanomyces bruxellensis population survey reveals a diploid-triploid complex structured according to substrate of isolation and geographical distribution. Sci Rep 8(1):1–13CrossRefGoogle Scholar
  4. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72(1–2):248–254CrossRefGoogle Scholar
  5. Branco P, Francisco D, Chambon C, Hébraud M, Arneborg N et al (2014) Identification of novel GAPDH-derived antimicrobial peptides secreted by Saccharomyces cerevisiae and involved in wine microbial interactions. Appl Microbiol Biotechnol 98(2):843–853CrossRefGoogle Scholar
  6. Branco P, Viana T, Albergaria H, Arneborg N (2015) Antimicrobial peptides (AMPs) produced by Saccharomyces cerevisiae induce alterations in the intracellular pH, membrane permeability and culturability of Hanseniaspora guilliermondii cells. Int J Food Microbiol 205:112–118CrossRefGoogle Scholar
  7. Branco P, Francisco D, Monteiro M, Almeida MG, Caldeira J et al (2017) Antimicrobial properties and death-inducing mechanisms of saccharomycin, a biocide secreted by Saccharomyces cerevisiae. Appl Microbiol Biotechnol 101(1):159–171CrossRefGoogle Scholar
  8. Comitini F, De Ingeniis J, Pepe L, Mannazzu I, Ciani M (2004) Pichia anomala and Kluyveromyces wickerhamii killer toxins as new tools against Dekkera/Brettanomyces spoilage yeasts. FEMS Microbiol Lett 238(1):235–240CrossRefGoogle Scholar
  9. Coronado P, Aguilera S, Carmona L, Godoy L, Martínez C et al (2015) Comparison of the behaviour of Brettanomyces bruxellensis strain LAMAP L2480 growing in authentic and synthetic wines. Antonie Leeuwenhoek 107(5):1217–1223CrossRefGoogle Scholar
  10. Curtin CD, Borneman AR, Chambers PJ, Pretorius IS (2012) De-novo assembly and analysis of the heterozygous triploid genome of the wine spoilage yeast Dekkera bruxellensis AWRI1499. PLoS ONE 7(3):1–10CrossRefGoogle Scholar
  11. Devalia JL, Rusznak C, Herdman MJ, Trigg CJ, Davies RJ et al (1994) Effect of nitrogen dioxide and sulphur dioxide on airway response of mild asthmatic patients to allergen inhalation. Lancet 344(8938):1668–1671CrossRefGoogle Scholar
  12. Enrique M, Marcos JF, Yuste M, Martínez M, Vallés S et al (2007) Antimicrobial action of synthetic peptides towards wine spoilage yeasts. Int J Food Microbiol 118(3):318–325CrossRefGoogle Scholar
  13. Fabrizio V, Vigentini I, Parisi N, Picozzi C, Compagno C et al (2015) Heat inactivation of wine spoilage yeast Dekkera bruxellensis by hot water treatment. Lett Appl Microbiol 61(2):186–191CrossRefGoogle Scholar
  14. Ganga MA, Martínez C (2004) Effect of wine yeast monoculture practice on the biodiversity of non-Saccharomyces yeasts. J Appl Microbiol 96(1):76–83CrossRefGoogle Scholar
  15. Godoy L, Vera-Wolf P, Martinez C, Ugalde JA, Ganga MA (2016) Comparative transcriptome assembly and genome-guided profiling for Brettanomyces bruxellensis LAMAP2480 during p-coumaric acid stress. Sci Rep 6:1–13CrossRefGoogle Scholar
  16. Godoy L, Silva-Moreno E, Mardones W, Guzman D, Cubillos FA et al (2017) Genomics perspectives on metabolism, survival strategies, and biotechnological applications of Brettanomyces bruxellensis LAMAP2480. J Mol Microbiol Biotechnol 27(3):147–158CrossRefGoogle Scholar
  17. González-Arenzana L, Sevenich R, Rauh C, López R, Knorr D et al (2016) Inactivation of Brettanomyces bruxellensis by high hydrostatic pressure technology. Food Control 59:188–195CrossRefGoogle Scholar
  18. Hellborg L, Piškur J (2009) Complex nature of the genome in a wine spoilage yeast, Dekkera bruxellensis. Eukaryot Cell 8(11):1739–1749CrossRefGoogle Scholar
  19. Liti G, Carter DM, Moses AM, Warringer J, Parts L et al (2009) Population genomics of domestic and wild yeasts. Nature 458(7236):337–341CrossRefGoogle Scholar
  20. Matsuzaki K (2009) Control of cell selectivity of antimicrobial peptides. Biochim Biophys Acta 1788(8):1687–1692CrossRefGoogle Scholar
  21. Mehlomakulu NN, Setati ME, Divol B (2014) Characterization of novel killer toxins secreted by wine-related non-Saccharomyces yeasts and their action on Brettanomyces spp. Int J Food Microbiol 188:83–91CrossRefGoogle Scholar
  22. Mehlomakulu NN, Prior KJ, Setati ME, Divol B (2017) Candida pyralidae killer toxin disrupts the cell wall of Brettanomyces bruxellensis in red grape juice. J Appl Microbiol 122(3):747–758CrossRefGoogle Scholar
  23. Narayanan TK, Rao GR (1976) Beta-indoleethanol and beta-indolelactic acid production by Candida species: their antibacterial and autoantibiotic action. Antimicrob Agents Chemother 9(3):375–380CrossRefGoogle Scholar
  24. Oelofse A, Pretorius IS, du Toit M (2008) Significance of Brettanomyces and Dekkera during winemaking: a synoptic review. S Afr J Enol Vitic 29(2):128–144Google Scholar
  25. Piškur J, Ling Z, Marcet-Houben M, Ishchuk OP, Aerts A et al (2012) The genome of wine yeast Dekkera bruxellensis provides a tool to explore its food-related properties. Int J Food Microbiol 157(2):202–209CrossRefGoogle Scholar
  26. Puértolas E, López N, Condón S, Raso J, Álvarez I (2009) Pulsed electric fields inactivation of wine spoilage yeast and bacteria. Int J Food Microbiol 130(1):49–55CrossRefGoogle Scholar
  27. Roostita LB, Fleet GH, Wendry SP, Apon ZM, Gemilang LU (2011) Determination of yeasts antimicrobial activity in milk and meat products. Adv J Food Sci Technol 3(6):442–445Google Scholar
  28. Schägger H (2006) Tricine-SDS-PAGE. Nat Protocols 1(1):16–22CrossRefGoogle Scholar
  29. Schägger H, von Jagow G (1987) Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem 166(2):368–379CrossRefGoogle Scholar
  30. Valdes J, Tapia P, Cepeda V, Varela J, Godoy L et al (2014) Draft genome sequence and transcriptome analysis of the wine spoilage yeast Dekkera bruxellensis LAMAP2480 provides insights into genetic diversity, metabolism and survival. FEMS Microbiol Lett 361(2):104–106CrossRefGoogle Scholar
  31. Vigentini I, Lucy Joseph CM, Picozzi C, Foschino R, Bisson LF (2013) Assessment of the Brettanomyces bruxellensis metabolome during sulphur dioxide exposure. FEMS Yeast Res 13(7):597–608CrossRefGoogle Scholar
  32. Warringer J, Zörgö E, Cubillos FA, Zia A, Gjuvsland A et al (2011) Trait variation in yeast is defined by population history. PLoS Genet 7(6):e1002111. CrossRefGoogle Scholar
  33. Younis G, Awad A, Dawod RE, Yousef NE (2017) Antimicrobial activity of yeasts against some pathogenic bacteria. Vet World 10(8):979–983CrossRefGoogle Scholar
  34. Zhang L, Gallo RL (2016) Antimicrobial peptides. Curr Biol 26(1):R14–R19CrossRefGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2018

Authors and Affiliations

  1. 1.Laboratory of Biotechnology and Applied Microbiology, Department of Food Science and TechnologyUniversidad de Santiago de ChileSantiagoChile

Personalised recommendations