Biotechnological exploitation of Tetrapisispora phaffii killer toxin: heterologous production in Komagataella phaffii (Pichia pastoris)
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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Yeast killer toxins have mainly been studied with respect to the control of spoilage yeasts in the beverage and food industry or for medical applications.
“Yeast killer toxins have thus been mainly studied with respect to the control of spoilage yeasts in the beverage and food industry or for medical applications (Chessa et al. 2017; Chi et al. 2010; Lowes et al. 2000; Mannazzu et al. 2019; Schmitt and Breinig 2002).”
From Biocontrol yeasts: mechanisms and applications (2019) · cites this paper as Chessa et al. 2017
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- Authors
- Chessa, R., Landolfo, S., Ciani, M., Budroni, M., Zara, S., Ustun, M., Cakar, Z. P., & Mannazzu, I.
- Affiliations
- University of Sassari (IT)
- Marche Polytechnic University (IT)
- Corresponding author
- Mannazzu, Ilaria
- Venue
- Applied Microbiology and Biotechnology
- Published
- 2016-12-28
- Volume
- 101
- Issue
- 7
- Pages
- 2931-2942
- ISSN
- 0175-7598 (ISSN-L)
- 1432-0614
- OpenAlex
- W2567003690
- Cited by
- 22
- References
- 57
- Citations by year
- 2026: 2 · 2025: 1 · 2024: 4 · 2023: 2 · 2022: 1 · 2021: 2 · 2020: 3 · 2019: 3 · 2018: 2 · 2017: 2
- FWCI
- 1.21
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Applied Microbiology and Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- UnissResearch (Università degli Studi di Sassari) (submittedVersion)
- Università Politecnica delle Marche (Università Politecnica delle Marche) (submittedVersion)
- Istanbul Technical University Academic Open Archive (Istanbul Technical University) (submittedVersion)
- Funders
- Fondazione Banco di Sardegna (grant number 2014.0470)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Yeasts and Rust Fungi Studies
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Enzyme Production and Characterization
- Keywords
- killer toxin, Pichia pastoris, heterologous expression, recombinant protein expression, β-1,3-glucanase, natural antimicrobials, spoilage yeasts, Dekkera bruxellensis, wine spoilage, grape must
- MeSH terms
- Biotechnology, Cell Wall, Cytotoxins, Saccharomycetales, Kluyveromyces, Pichia, Recombinant Proteins, Saccharomyces cerevisiae, Wine, Yeasts, Zygosaccharomyces, Killer Factors, Yeast
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