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Biotechnological exploitation of Tetrapisispora phaffii killer toxin: heterologous production in Komagataella phaffii (Pichia pastoris)

  • Biotechnologically relevant enzymes and proteins
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Abstract

The use of natural antimicrobials from plants, animals and microorganisms to inhibit the growth of pathogenic and spoilage microorganisms is becoming more frequent. This parallels the increased consumer interest towards consumption of minimally processed food and ‘greener’ food and beverage additives. Among the natural antimicrobials of microbial origin, the killer toxin produced by the yeast Tetrapisispora phaffii, known as Kpkt, appears to be a promising natural antimicrobial agent. Kpkt is a glycoprotein with β-1,3-glucanase and killer activity, which induces ultrastructural modifications to the cell wall of yeast of the genera Kloeckera/Hanseniaspora and Zygosaccharomyces. Moreover, Kpkt maintains its killer activity in grape must for at least 14 days under winemaking conditions, thus suggesting its use against spoilage yeast in wine making and the sweet beverage industry. Here, the aim was to explore the possibility of high production of Kpkt for biotechnological exploitation. Molecular tools for heterologous production of Kpkt in Komagataella phaffii GS115 were developed, and two recombinant clones that produce up to 23 mg/L recombinant Kpkt (rKpkt) were obtained. Similar to native Kpkt, rKpkt has β-glucanase and killer activities. Moreover, it shows a wider spectrum of action with respect to native Kpkt. This includes effects on Dekkera bruxellensis, a spoilage yeast of interest not only in wine making, but also for the biofuel industry, thus widening the potential applications of this rKpkt.

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Acknowledgments

This study and S.L. were financially supported by Fondazione Banco di Sardegna with the project named ‘Produzione eterologa di una glicoproteina di lievito da impiegare come antimicrobico naturale nell’industria enologica e delle bevande’ (grant number 2014.0470 to IM). M.U. was financially supported by the ERASMUS Exchange Program between the University of Sassari and Istanbul Technical University.

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Authors and Affiliations

  1. Department of Agriculture, University of Sassari, Sassari, Italy

    Rossella Chessa, Sara Landolfo, Marilena Budroni, Severino Zara, Murat Ustun & Ilaria Mannazzu

  2. Department of Life and Environmental Sciences, Polytechnic University of Marche, Ancona, Italy

    Maurizio Ciani

  3. Enology and Viticulture Research Group, University of Sassari, Sassari, Italy

    Marilena Budroni, Severino Zara & Ilaria Mannazzu

  4. Department of Molecular Biology and Genetics, Faculty of Science & Letters, Istanbul Technical University, Istanbul, Turkey

    Murat Ustun & Zeynep Petek Cakar

  5. Dr. Orhan Ocalgiray Molecular Biology, Biotechnology and Genetics Research Center (ITU-MOBGAM), Istanbul Technical University, Istanbul, Turkey

    Murat Ustun & Zeynep Petek Cakar

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  1. Rossella Chessa
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  2. Sara Landolfo
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  3. Maurizio Ciani
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Correspondence to Ilaria Mannazzu.

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This study was funded by Fondazione Banco di Sardegna (grant number 2014.0470).

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Chessa, R., Landolfo, S., Ciani, M. et al. Biotechnological exploitation of Tetrapisispora phaffii killer toxin: heterologous production in Komagataella phaffii (Pichia pastoris) . Appl Microbiol Biotechnol 101, 2931–2942 (2017). https://doi.org/10.1007/s00253-016-8050-2

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