Purification and Characterization of WA18, a New Mycocin Produced by <i>Wickerhamomyces anomalus</i> Active in Wine Against <i>Brettanomyces bruxellensis</i> Spoilage Yeasts
<i>Wickerhamomyces anomalus</i> strain 18, isolated from a natural underground cheese ripening pit, secretes a mycocin named WA18 that inhibits wine spoilage yeasts belonging to <i>Brettanomyces bruxellensis</i> species, with a broad-spectrum of activity. WA18 was purified, a...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-12-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/9/1/56 |
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| author | Francesca Comitini Alice Agarbati Laura Canonico Edoardo Galli Maurizio Ciani |
| author_facet | Francesca Comitini Alice Agarbati Laura Canonico Edoardo Galli Maurizio Ciani |
| author_sort | Francesca Comitini |
| collection | DOAJ |
| description | <i>Wickerhamomyces anomalus</i> strain 18, isolated from a natural underground cheese ripening pit, secretes a mycocin named WA18 that inhibits wine spoilage yeasts belonging to <i>Brettanomyces bruxellensis</i> species, with a broad-spectrum of activity. WA18 was purified, and the purified protein was digested with specific restriction enzymes (lysine K and arginine R cut sites). The LC–MS and LC–MS/MS analysis after enzymatic digestions revealed a molecular weight of 31 kDa. Bioinformatics processing and database research of digested pure killer protein showed 99% identity with a UDP-glycosyltransferase protein. Competitive inhibition assay of killer activity by cell-wall polysaccharides suggests that branched glucans represent the first receptor site of the toxin on the envelope of the sensitive target. The WA18 partially purified crude extract (PPCE) showed high stability of antimicrobial activity at the physicochemical conditions suitable for the winemaking process. Indeed, in wine WA18 was able to counteract <i>B. bruxellensis</i> and control the production of ethyl phenols. In addition, the strain WA18 was compatible with <i>Saccharomyces cerevisiae</i> in co-culture conditions with a potential application together with commercial starter cultures. These data suggest that WA18 mycocin is a promising biocontrol agent against spoilage yeasts in winemaking, particularly during wine storage. |
| first_indexed | 2024-03-10T13:43:44Z |
| format | Article |
| id | doaj.art-9cde354b535c4c3ab8c617b70ef44ec6 |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-10T13:43:44Z |
| publishDate | 2020-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-9cde354b535c4c3ab8c617b70ef44ec62023-11-21T02:47:26ZengMDPI AGMicroorganisms2076-26072020-12-01915610.3390/microorganisms9010056Purification and Characterization of WA18, a New Mycocin Produced by <i>Wickerhamomyces anomalus</i> Active in Wine Against <i>Brettanomyces bruxellensis</i> Spoilage YeastsFrancesca Comitini0Alice Agarbati1Laura Canonico2Edoardo Galli3Maurizio Ciani4Dipartimento di Scienze della Vita e dell’Ambiente (DiSVA), Università Politecnica delle Marche, Via Brecce Bianche, 60131 Ancona, ItalyDipartimento di Scienze della Vita e dell’Ambiente (DiSVA), Università Politecnica delle Marche, Via Brecce Bianche, 60131 Ancona, ItalyDipartimento di Scienze della Vita e dell’Ambiente (DiSVA), Università Politecnica delle Marche, Via Brecce Bianche, 60131 Ancona, ItalyDipartimento di Scienze della Vita e dell’Ambiente (DiSVA), Università Politecnica delle Marche, Via Brecce Bianche, 60131 Ancona, ItalyDipartimento di Scienze della Vita e dell’Ambiente (DiSVA), Università Politecnica delle Marche, Via Brecce Bianche, 60131 Ancona, Italy<i>Wickerhamomyces anomalus</i> strain 18, isolated from a natural underground cheese ripening pit, secretes a mycocin named WA18 that inhibits wine spoilage yeasts belonging to <i>Brettanomyces bruxellensis</i> species, with a broad-spectrum of activity. WA18 was purified, and the purified protein was digested with specific restriction enzymes (lysine K and arginine R cut sites). The LC–MS and LC–MS/MS analysis after enzymatic digestions revealed a molecular weight of 31 kDa. Bioinformatics processing and database research of digested pure killer protein showed 99% identity with a UDP-glycosyltransferase protein. Competitive inhibition assay of killer activity by cell-wall polysaccharides suggests that branched glucans represent the first receptor site of the toxin on the envelope of the sensitive target. The WA18 partially purified crude extract (PPCE) showed high stability of antimicrobial activity at the physicochemical conditions suitable for the winemaking process. Indeed, in wine WA18 was able to counteract <i>B. bruxellensis</i> and control the production of ethyl phenols. In addition, the strain WA18 was compatible with <i>Saccharomyces cerevisiae</i> in co-culture conditions with a potential application together with commercial starter cultures. These data suggest that WA18 mycocin is a promising biocontrol agent against spoilage yeasts in winemaking, particularly during wine storage.https://www.mdpi.com/2076-2607/9/1/56killer yeastskiller toxin<i>Wickerhamomyces anomalus</i>spoilage yeastsbiocontrolbioactive yeasts |
| spellingShingle | Francesca Comitini Alice Agarbati Laura Canonico Edoardo Galli Maurizio Ciani Purification and Characterization of WA18, a New Mycocin Produced by <i>Wickerhamomyces anomalus</i> Active in Wine Against <i>Brettanomyces bruxellensis</i> Spoilage Yeasts Microorganisms killer yeasts killer toxin <i>Wickerhamomyces anomalus</i> spoilage yeasts biocontrol bioactive yeasts |
| title | Purification and Characterization of WA18, a New Mycocin Produced by <i>Wickerhamomyces anomalus</i> Active in Wine Against <i>Brettanomyces bruxellensis</i> Spoilage Yeasts |
| title_full | Purification and Characterization of WA18, a New Mycocin Produced by <i>Wickerhamomyces anomalus</i> Active in Wine Against <i>Brettanomyces bruxellensis</i> Spoilage Yeasts |
| title_fullStr | Purification and Characterization of WA18, a New Mycocin Produced by <i>Wickerhamomyces anomalus</i> Active in Wine Against <i>Brettanomyces bruxellensis</i> Spoilage Yeasts |
| title_full_unstemmed | Purification and Characterization of WA18, a New Mycocin Produced by <i>Wickerhamomyces anomalus</i> Active in Wine Against <i>Brettanomyces bruxellensis</i> Spoilage Yeasts |
| title_short | Purification and Characterization of WA18, a New Mycocin Produced by <i>Wickerhamomyces anomalus</i> Active in Wine Against <i>Brettanomyces bruxellensis</i> Spoilage Yeasts |
| title_sort | purification and characterization of wa18 a new mycocin produced by i wickerhamomyces anomalus i active in wine against i brettanomyces bruxellensis i spoilage yeasts |
| topic | killer yeasts killer toxin <i>Wickerhamomyces anomalus</i> spoilage yeasts biocontrol bioactive yeasts |
| url | https://www.mdpi.com/2076-2607/9/1/56 |
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