Abstract
In our previous study, it was found that the killer toxin produced by the marine-derived yeast Wickerhamomyces anomalus YF07b has both killing activity and β-1,3-glucanase activity and the molecular mass of it is 47.0 kDa. In this study, the same yeast strain was found to produce another killer toxin which only had killing activity against some yeast strains, but had no β-1,3-glucanase activity and the molecular mass of the purified killer toxin was 67.0 kDa. The optimal pH, temperature and NaCl concentration for action of the purified killer toxin were 3.5, 16 °C and 4.0 % (w/v), respectively. The purified killer toxin could be bound by the whole sensitive yeast cells, but was not bound by manann, chitin and β-1,3-glucan. The purified killer toxin had killing activity against Yarrowia lipolytica, Saccharomyces cerevisiae, Metschnikowia bicuspidata WCY, Candida tropicalis, Candida albicans and Kluyveromyces aestuartii. Lethality of the sensitive cells treated by the newly purified killer toxin from W. anomalus YF07b involved disruption of cellular integrity by permeabilizing cytoplasmic membrane function.
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References
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 72:248–254
Buzdar MA, Chi Z, Wang Q, Hua MX, Chi ZM (2011) Production, purification, and characterization of a novel killer toxin from Kluyveromyces siamensis against a pathogenic yeast in crab. Appl Microbiol Biotechnol 91:1571–1579
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 238:235–240
De Ingeniis J, Raffaelli N, Ciani M, Mannazzu I (2009) Pichia anomala DBVPG 3003 secretes a ubiquitin-like protein that has antimicrobial activity. Appl Environ Microbiol 75:1129–1134
Gunge N, Tamaru A, Ozawa F, Sakaguchi K (1981) Isolation and characterization of linear deoxyribonucleic acid plasmids from Kluyveromyces lactis and the plasmid-associated killer character. J Bacteriol 145:382–390
Guyard C, Dehecq E, Tissier JP, Polonelli L, Dei-Cas E, Cailliez JC, Menozzi FD (2002) Involvement of β-glucans in the widespectrum antimicrobial activity of Williopsis saturnus var. mrakii MUCL 41968 killer toxin. Mol Med 8:686–694
Hua MX, Chi Z, Liu GL, Buzdar MA, Chi ZM (2010) Production of a novel and old-active killer toxin by Mrakia frigida 2E00797 isolated from sea sediment in Antarctica. Extremophiles 14:515–521
Kim J, Subbery P (2011) Candida albicans, a major human fungal pathogen. J Microbiol 49:171–177
Laemmli UK (1970) Cleavage of structural proteins during assembly of head of bacteriophage T4. Nature 227:680–685
Liu GL, Ke 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 47:822–827
Magliani W, Conti S, Travassos LR, Polonelli L (2008) From yeast killer toxins to antibiobodies and beyond. FEMS Microbiol Lett 288:1–8
Peng Y, Chi ZM, Wang XH, Li J (2010) β-1,3-glucanase inhibits activity of the killer toxin produced by the marine-derived yeast Williopsis saturnus WC91-2. Mar Biotechnol 12:479–485
Santos A, Marquina D (2004) Ion channel activity by Pichia membranifaciens killer toxin. Yeast 21:151–162
Santos A, Marquina D, Leal JA, Peinado JM (2000) (1 → 6)-beta-D’glucan as cell wall receptor for Pichia membranifaciens killer toxin. Appl Environ Microbiol 66:1809–1813
Schmitt MJ, Breinig F (2002) The viral killer system in yeast: from molecular biology to application. FEMS Microbiol Rev 26:257–276
Spiro RG (1966) Analysis of sugars found in glycoproteins. Meth Enzymol 8:3–26
Sun HY, Wang K, Chi Z, Xu HM, Chi ZM (2012) Simultaneous production of single cell protein and killer toxin by Wickerhamomyces anomalus HN1-2 isolated from mangrove ecosystem. Proc Biochem 47:251–256
Varghese G, Diwan AM (1983) Simultaneous staining of proteins during polyacrylamide gel electrophoresis in acidic gels by countermigration of Coomassie brilliant blue R-250. Anal Biochem 132:481–483
Wang X, Chi ZM, Yue L, Li J, Li M, Wu L (2007a) A marine killer yeast against the pathogenic yeast strain in crab (Portunus trituberculatus) and an optimization of the toxin production. Microbiol Res 162:77–85
Wang XH, Chi ZM, Yue L, Li J (2007b) Purification and characterization of killer toxin from a marine yeast Pichia anomala YF07b against the pathogenic yeast in crab. Curr Microbiol 55:396–401
Wang L, Yue L, Chi ZM, Wang X (2008) Marine killer yeasts active against a yeast strain pathogenic to crab Portunus trituberculatus. Dis Aquat Organ 80:211–218
Wang XX, Chi Z, Peng Y, Wang XH, Ru SG, Chi ZM (2011) Purification, characterization and gene cloning of the killer toxin produced by the marine-derived yeast Williopsis saturnus WC91-2. Microbiol Res. doi:10.1016/j.micres.2011.12.001
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This work was supported by grant 30771645 from the National Natural Science Foundation of China.
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Guo, FJ., Ma, Y., Xu, HM. et al. A novel killer toxin produced by the marine-derived yeast Wickerhamomyces anomalus YF07b. Antonie van Leeuwenhoek 103, 737–746 (2013). https://doi.org/10.1007/s10482-012-9855-3
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DOI: https://doi.org/10.1007/s10482-012-9855-3