Evaluation of Recombinant Kpkt Cytotoxicity on HaCaT Cells: Further Steps towards the Biotechnological Exploitation Yeast Killer Toxins
The soil yeast <i>Tetrapisispora phaffii</i> secretes a killer toxin, named Kpkt, that shows β-glucanase activity and is lethal to wine spoilage yeasts belonging to <i>Kloeckera/Hanseniaspora</i>, <i>Saccharomycodes</i> and <i>Zygosaccharomyces</i>. Wh...
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MDPI AG
2021-03-01
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| Series: | Foods |
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| Online Access: | https://www.mdpi.com/2304-8158/10/3/556 |
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| author | Gavino Carboni Ivana Marova Giacomo Zara Severino Zara Marilena Budroni Ilaria Mannazzu |
| author_facet | Gavino Carboni Ivana Marova Giacomo Zara Severino Zara Marilena Budroni Ilaria Mannazzu |
| author_sort | Gavino Carboni |
| collection | DOAJ |
| description | The soil yeast <i>Tetrapisispora phaffii</i> secretes a killer toxin, named Kpkt, that shows β-glucanase activity and is lethal to wine spoilage yeasts belonging to <i>Kloeckera/Hanseniaspora</i>, <i>Saccharomycodes</i> and <i>Zygosaccharomyces</i>. When expressed in <i>Komagataella phaffii</i>, recombinant Kpkt displays a wider spectrum of action as compared to its native counterpart, being active on a vast array of wine yeasts and food-related bacteria. Here, to gather information on recombinant Kpkt cytotoxicity, lyophilized preparations of this toxin (LrKpkt) were obtained and tested on immortalized human keratinocyte HaCaT cells, a model for the stratified squamous epithelium of the oral cavity and esophagus. LrKpkt proved harmless to HaCaT cells at concentrations up to 36 AU/mL, which are largely above those required to kill food-related yeasts and bacteria in vitro (0.25–2 AU/mL). At higher concentrations, it showed a dose dependent effect that was comparable to that of the negative control and therefore could be ascribed to compounds, other than the toxin, occurring in the lyophilized preparations. Considering the dearth of studies regarding the effects of yeast killer toxins on human cell lines, these results represent a first mandatory step towards the evaluation the possible risks associated to human intake. Moreover, in accordance with that observed on <i>Ceratitis capitata</i> and <i>Musca domestica</i>, they support the lack of toxicity of this toxin on non-target eukaryotic models and corroborate the possible exploitation of killer toxins as natural antimicrobials in the food and beverages industries. |
| first_indexed | 2024-03-09T05:04:03Z |
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| issn | 2304-8158 |
| language | English |
| last_indexed | 2024-03-09T05:04:03Z |
| publishDate | 2021-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Foods |
| spelling | doaj.art-6c4d6d94cd234fed80c9409f4b6dadd82023-12-03T12:57:14ZengMDPI AGFoods2304-81582021-03-0110355610.3390/foods10030556Evaluation of Recombinant Kpkt Cytotoxicity on HaCaT Cells: Further Steps towards the Biotechnological Exploitation Yeast Killer ToxinsGavino Carboni0Ivana Marova1Giacomo Zara2Severino Zara3Marilena Budroni4Ilaria Mannazzu5Department of Agricultural Sciences, University of Sassari, Viale Italia 39, 07100 Sassari, ItalyFaculty of Chemistry, Brno University of Technology, Purkyňova 464/118, Královo Pole, 61200 Brno, Czech RepublicDepartment of Agricultural Sciences, University of Sassari, Viale Italia 39, 07100 Sassari, ItalyDepartment of Agricultural Sciences, University of Sassari, Viale Italia 39, 07100 Sassari, ItalyDepartment of Agricultural Sciences, University of Sassari, Viale Italia 39, 07100 Sassari, ItalyDepartment of Agricultural Sciences, University of Sassari, Viale Italia 39, 07100 Sassari, ItalyThe soil yeast <i>Tetrapisispora phaffii</i> secretes a killer toxin, named Kpkt, that shows β-glucanase activity and is lethal to wine spoilage yeasts belonging to <i>Kloeckera/Hanseniaspora</i>, <i>Saccharomycodes</i> and <i>Zygosaccharomyces</i>. When expressed in <i>Komagataella phaffii</i>, recombinant Kpkt displays a wider spectrum of action as compared to its native counterpart, being active on a vast array of wine yeasts and food-related bacteria. Here, to gather information on recombinant Kpkt cytotoxicity, lyophilized preparations of this toxin (LrKpkt) were obtained and tested on immortalized human keratinocyte HaCaT cells, a model for the stratified squamous epithelium of the oral cavity and esophagus. LrKpkt proved harmless to HaCaT cells at concentrations up to 36 AU/mL, which are largely above those required to kill food-related yeasts and bacteria in vitro (0.25–2 AU/mL). At higher concentrations, it showed a dose dependent effect that was comparable to that of the negative control and therefore could be ascribed to compounds, other than the toxin, occurring in the lyophilized preparations. Considering the dearth of studies regarding the effects of yeast killer toxins on human cell lines, these results represent a first mandatory step towards the evaluation the possible risks associated to human intake. Moreover, in accordance with that observed on <i>Ceratitis capitata</i> and <i>Musca domestica</i>, they support the lack of toxicity of this toxin on non-target eukaryotic models and corroborate the possible exploitation of killer toxins as natural antimicrobials in the food and beverages industries.https://www.mdpi.com/2304-8158/10/3/556Kpktyeast killer toxinnatural antimicrobialHaCaT cells |
| spellingShingle | Gavino Carboni Ivana Marova Giacomo Zara Severino Zara Marilena Budroni Ilaria Mannazzu Evaluation of Recombinant Kpkt Cytotoxicity on HaCaT Cells: Further Steps towards the Biotechnological Exploitation Yeast Killer Toxins Foods Kpkt yeast killer toxin natural antimicrobial HaCaT cells |
| title | Evaluation of Recombinant Kpkt Cytotoxicity on HaCaT Cells: Further Steps towards the Biotechnological Exploitation Yeast Killer Toxins |
| title_full | Evaluation of Recombinant Kpkt Cytotoxicity on HaCaT Cells: Further Steps towards the Biotechnological Exploitation Yeast Killer Toxins |
| title_fullStr | Evaluation of Recombinant Kpkt Cytotoxicity on HaCaT Cells: Further Steps towards the Biotechnological Exploitation Yeast Killer Toxins |
| title_full_unstemmed | Evaluation of Recombinant Kpkt Cytotoxicity on HaCaT Cells: Further Steps towards the Biotechnological Exploitation Yeast Killer Toxins |
| title_short | Evaluation of Recombinant Kpkt Cytotoxicity on HaCaT Cells: Further Steps towards the Biotechnological Exploitation Yeast Killer Toxins |
| title_sort | evaluation of recombinant kpkt cytotoxicity on hacat cells further steps towards the biotechnological exploitation yeast killer toxins |
| topic | Kpkt yeast killer toxin natural antimicrobial HaCaT cells |
| url | https://www.mdpi.com/2304-8158/10/3/556 |
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