Advertisement

Antonie van Leeuwenhoek

, Volume 107, Issue 4, pp 961–970 | Cite as

A novel killer protein from Pichia kluyveri isolated from an Algerian soil: purification and characterization of its in vitro activity against food and beverage spoilage yeasts

  • Fatima-Zohra Kenza LabbaniEmail author
  • Benedetta Turchetti
  • Leila Bennamoun
  • Scheherazad Dakhmouche
  • Rita Roberti
  • Lanfranco Corazzi
  • Zahia Meraihi
  • Pietro Buzzini
Original Paper

Abstract

A novel killer protein (Pkkp) secreted by a Pichia kluyveri strain isolated from an Algerian soil was active against food and beverage spoilage yeasts of the genera Dekkera, Kluyveromyces, Pichia, Saccharomyces, Torulaspora, Wickerhamomyces and Zygosaccharomyces. After purification by gel filtration chromatography Pkkp revealed an apparent molecular mass of 54 kDa with SDS-PAGE. Minimum inhibitory concentrations (MICs) of purified Pkkp exhibited a high in vitro activity against Dekkera bruxellensis (MICs from 64,000- to 256,000-fold lower than that exhibited by potassium metabisulphite) and Saccharomyces cerevisiae (MICs from 32,000- to 64,000- fold lower than potassium sorbate). No in vitro synergistic interactions (calculated by FIC index − Σ FIC) were observed when Pkkp was used in combination with potassium metabisulphite, potassium sorbate, or ethanol. Pkkp exhibited a dose–response effect against D. bruxellensis and S. cerevisiae in a low-alcoholic drink and fruit juice, respectively. The results of the present study suggest that Pkkp could be proposed as a novel food-grade compound useful for the control of food and beverage spoilage yeasts.

Keywords

Yeast killer protein Pichia kluyveri Susceptibility testing Spoilage yeasts Minimum inhibitory concentrations (MICs) 

Notes

Acknowledgments

The author Fatima-Zohra Kenza Labbani is deeply grateful to the Ministry of Higher Education and Scientific Research of Algeria for the doctoral fellowship (PNE program).

Supplementary material

10482_2015_388_MOESM1_ESM.pdf (30 kb)
Supplementary material 1 (PDF 30 kb)
10482_2015_388_MOESM2_ESM.pdf (27 kb)
Supplementary material 2 (PDF 26 kb)
10482_2015_388_MOESM3_ESM.pdf (21 kb)
Supplementary material 3 (PDF 20 kb)
10482_2015_388_MOESM4_ESM.pdf (247 kb)
Supplementary material 4 (PDF 247 kb)
10482_2015_388_MOESM5_ESM.pdf (30 kb)
Supplementary material 5 (PDF 30 kb)
10482_2015_388_MOESM6_ESM.pdf (14 kb)
Supplementary material 6 (PDF 14 kb)
10482_2015_388_MOESM7_ESM.pdf (22 kb)
Supplementary material 7 (PDF 22 kb)

References

  1. Bajaj BK, Raina S, Singh S (2012) Killer toxin from a novel killer yeast Pichia kudriavzevii RY55 with idiosyncratic antibacterial activity. J Basic Microbiol 52:1–11CrossRefGoogle Scholar
  2. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-311 dye binding. Anal Biochem 72:248–254CrossRefPubMedGoogle Scholar
  3. Buzzini P, Corazzi L, Turchetti B, Buratta M, Martini A (2004) Characterization of the in vitro antimycotic activity of a novel killer protein from Williopsis saturnus DBVPG 4561 against emerging pathogenic yeasts. FEMS Microbiol Lett 238:359–365PubMedGoogle Scholar
  4. Ciani M, Fatichenti F (2001) Killer toxin of Kluyveromyces phaffii DBVPG 6076 as a biopreservative agent to control apiculate wine yeasts. Appl Environ Microbiol 67:3058–3063CrossRefPubMedCentralPubMedGoogle Scholar
  5. Clinical and Laboratory Standards Institute (2002) Reference method for broth dilution antifungal susceptibility testing of yeasts. Approved standard 2nd edn. NCCLS document M27-A2, Pennsylvania, USAGoogle Scholar
  6. Comitini F, Ciani M (2011) Kluyveromyces wickerhamii killer toxin: purification and activity towards Brettanomyces/Dekkera yeasts in grape must. FEMS Microbiol Lett 316:77–82CrossRefPubMedGoogle Scholar
  7. 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 238:235–240CrossRefPubMedGoogle Scholar
  8. Dabhole MP, Joishy KN (2005) Production and effect of killer toxin by Saccharomyces cerevisiae and Pichia kluyveri on sensitive yeasts and fungal pathogens. Indian J Biotechnol 4:290–292Google Scholar
  9. El-Benna AA, El-Sahn MA, Shehata MG (2011) Yeasts producing killer toxins: an overview. Alex J Food Sci Technol 8:41–53Google Scholar
  10. Golubev WI (2006) Antagonistic interactions among yeasts. In: Rosa CA, Péter G (eds) Biodiversity and ecophysiology of yeasts. Springer, Berlin, pp 197–219CrossRefGoogle Scholar
  11. Goretti M, Turchetti B, Buratta M, Branda E, Corazzi L, Vaughan-Martini A, Buzzini P (2009) In vitro antimycotic activity of a Williopsis saturnus killer protein against food spoilage yeasts. Int J Food Microbiol 131:178–182CrossRefPubMedGoogle Scholar
  12. Guo FJ, Ma Y, Xu HM, Wang XH, Chi ZM (2013) A novel killer toxin produced by the marine-derived yeast Wickerhamomyces anomalus YF07b. Antonie Van Leeuwenhoek 103:737–746. doi: 10.1007/s10482-012-9855-3 CrossRefPubMedGoogle Scholar
  13. Hodgson VJ, Button D, Walker GM (1995) Anti-Candida activity of a novel killer toxin from the yeast Williopsis mrakii. Microbiology 141:2003–2012CrossRefPubMedGoogle Scholar
  14. İzgü F, Altinbay D, Türeli AE (2007) In vitro susceptibilities of Candida spp. to Panomycocin, a novel exo-b-1,3-glucanase isolated from Pichia anomala NCYC 434. Microbiol Immunol 51:797–803CrossRefPubMedGoogle Scholar
  15. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefPubMedGoogle Scholar
  16. Liu SQ, Tsao M (2009) Inhibition of spoilage yeasts in cheese by killer yeast Williopsis saturnus var. saturnus. Int J Food Microbiol 131:280–282CrossRefPubMedGoogle Scholar
  17. Liu GL, Wang K, Hua MX, Buzdar MA, Chi ZM (2012) Purification and characterization of the cold-active killer toxin from the psychrotolerant yeast Mrakia frigida isolated from sea sediments in Antarctica. Proc Biochem 47:822–827CrossRefGoogle Scholar
  18. Magliani W, Conti S, Gerloni M, Bertolotti D, Polonelli L (1997) Yeast killer systems. Clin Microbiol Rev 10:369–400PubMedCentralPubMedGoogle Scholar
  19. Maráz A, Kovács M (2014) Food spoilage by cold-adapted yeasts. In: Buzzini P, Margesin R (eds) cold-adapted yeasts. Springer, Berlin, pp 497–532CrossRefGoogle Scholar
  20. Marquina D, Santos A, Peinado JM (2002) Biology of killer yeasts. Int Microbiol 5:65–71CrossRefPubMedGoogle Scholar
  21. Middelbeek EJ, Hermans JMH, Stumm C (1979) Production, purification and properties of a Pichia kluyveri killer toxin. Antonie Van Leeuwenhoek 45:437–450CrossRefPubMedGoogle Scholar
  22. Middelbeek EJ, Stumm C, Vogels GD (1980) Effects of a Pichia kluyveri killer toxin on sensitive cells. Antonie Van Leeuwenhoek 46:205–220CrossRefPubMedGoogle Scholar
  23. Odds FC (2003) Synergy, antagonism, and what the chequerboard put between them. J Antimicrob Chemother 52:1CrossRefPubMedGoogle Scholar
  24. Papadimitriou MNB, Resende C, Kuchler K, Brul S (2007) High Pdr12 levels in spoilage yeast (Saccharomyces cerevisiae) correlate directly with sorbic acid levels in the culture medium but are not sufficient to provide cells with acquired resistance to the food preservative. Int J Food Microbiol 113:173–179CrossRefPubMedGoogle Scholar
  25. Pillai SK, Moellering RC, Eliopoulos GM (2005) Antimicrobial combination. In: Lorian V (ed) Antibiotics in laboratory medicine. Williams and Wilkins, Philadelphia, pp 365–424Google Scholar
  26. Polonelli L, Magliani W, Ciociola T, Giovati L, Conti S (2011) From Pichia anomala killer toxin through killer antibodies to killer peptides for a comprehensive anti-infective strategy. Antonie Van Leeuwenhoek 99:35–41CrossRefPubMedGoogle Scholar
  27. Santos A, Marquina D (2004) Killer toxin of Pichia membranifaciens and its possible use as a biopreservative agent to control grey mould disease of grapevine. Microbiology 150:2527–2534CrossRefPubMedGoogle Scholar
  28. Santos A, San Mauro M, Bravo E, Marquina D (2009) PMKT2, a new killer toxin from Pichia membranifaciens, and its promising biotechnological properties for control of the spoilage yeast Brettanomyces bruxellensis. Microbiology 155:624–634CrossRefPubMedGoogle Scholar
  29. Santos A, Navascués E, Bravo E, Marquina D (2011) Ustilago maydis killer toxin as a new tool for the biocontrol of the wine spoilage yeast Brettanomyces bruxellensis. Int J Food Microbiol 145:147–154CrossRefPubMedGoogle Scholar
  30. Starmer WT, Ganter PF, Aberdeen V (1992) Geographic distribution and denetics of killer phenotypes for the yeast Pichia kluyveri across the United States. Appl Environ Microbiol 58:990–997PubMedCentralPubMedGoogle Scholar
  31. Stratford M (2006) Food and beverage spoilage yeasts. In: Querol A, Fleet GH (eds) Yeasts in Food and Beverage. Springer, Berlin, pp 335–379CrossRefGoogle Scholar
  32. Tserennadmid R, Takó M, Galgóczy L, Papp T, Pesti M, Vágvölgyi C, Almássy K, Krisch J (2011) Anti yeast activities of some essential oils in growth medium, fruit juices and milk. Int J Food Microbiol 144:480–486CrossRefPubMedGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  • Fatima-Zohra Kenza Labbani
    • 1
    • 2
    Email author
  • Benedetta Turchetti
    • 3
  • Leila Bennamoun
    • 4
  • Scheherazad Dakhmouche
    • 4
  • Rita Roberti
    • 5
  • Lanfranco Corazzi
    • 5
  • Zahia Meraihi
    • 4
  • Pietro Buzzini
    • 3
  1. 1.Department of Molecular and Cellular Biology, Natural and Life Sciences FacultyAbbes Laghrour University of KhenchelaKhenchelaAlgeria
  2. 2.Department of BiochemistryNatural and Life Sciences Faculty, University of Constantine 1ConstantineAlgeria
  3. 3.Department of Agricultural, Environmental and Food Science & Industrial Yeasts Collection DBVPGUniversity of PerugiaPerugiaItaly
  4. 4.Department of Biochemistry, Natural and Life Sciences FacultyUniversity of Constantine 1ConstantineAlgeria
  5. 5.Department of Experimental MedicineUniversity of PerugiaPerugiaItaly

Personalised recommendations