Biocontrol of Brettanomyces/Dekkera bruxellensis in alcoholic fermentations using saccharomycin-overproducing Saccharomyces cerevisiae strains
Biocontrol of Brettanomyces/Dekkera bruxellensis in alcoholic fermentations using saccharomycin-overproducing Saccharomyces cerevisiae strains
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DOI：
10.1007/s00253-019-09657-7
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发表时间：
2019-04-01
影响因子：
5
通讯作者：
Prista, Catarina
中科院分区：
文献类型：
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作者：
Branco, Patricia;Sabir, Farzana;Prista, Catarina
酒精发酵过程（例如酿酒和燃料乙醇生产）的微生物污染对于工业来说是一个严重的问题，因为它可能使产品不可接受和/或降低其生产率，导致巨大的经济损失。布鲁塞尔酒香酵母（Brettanomyces/Dekkera bruxellensis）是乙醇工业发酵过程中最危险的微生物污染物之一。在葡萄酒的情况下，这种酵母可以产生酚类化合物，赋予异味的最终产品。在燃料乙醇发酵中，D. Bruxellensis是影响乙醇产率和生产率的持久性污染物。我们最近发现，酿酒酵母分泌的杀生物剂，我们命名为糖霉素，由来自糖酵解酶甘油醛-3-磷酸脱氢酶（GAPDH）的抗菌肽（AMP）。阿托霉素对几种与葡萄酒相关的酵母菌有活性，即D。布鲁塞尔人然而，由S.酿酒酵母在酒精发酵过程中的死亡率不足以确保D.布鲁塞尔人因此，本工作的目的是构建转基因S。酿酒酵母菌株过量产生这些GAPDH衍生的AMP。在修饰的S.通过RT-qPCR对酿酒酵母菌株进行重组，证实了重组方法的成功。免疫学试验证实，经修饰的S。与未修饰的菌株相比，酿酒酵母菌株分泌更高量的AMP，诱导D.在酒精发酵过程中的布鲁塞尔。
Microbial contamination of alcoholic fermentation processes (e.g. winemaking and fuel-ethanol production) is a serious problem for the industry since it may render the product unacceptable and/or reduce its productivity, leading to large economic losses. Brettanomyces/Dekkera bruxellensis is one of the most dangerous microbial contaminant of ethanol industrial fermentations. In the case of wine, this yeast species can produce phenolic compounds that confer off-flavours to the final product. In fuel-ethanol fermentations, D. bruxellensis is a persistent contaminant that affects ethanol yields and productivities. We recently found that Saccharomyces cerevisiae secretes a biocide, which we named saccharomycin, composed of antimicrobial peptides (AMPs) derived from the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Saccharomycin is active against several wine-related yeast species, namely D. bruxellensis. However, the levels of saccharomycin naturally secreted by S. cerevisiae during alcoholic fermentation are not sufficient to ensure the complete death of D. bruxellensis. Therefore, the aim of the present work was to construct genetically modified S. cerevisiae strains to overproduce these GAPDH-derived AMPs. The expression levels of the nucleotides sequences encoding the AMPs were evaluated in the modified S. cerevisiae strains by RT-qPCR, confirming the success of the recombinant approach. Furthermore, we confirmed by immunological tests that the modified S. cerevisiae strains secreted higher amounts of the AMPs by comparison with the non-modified strain, inducing total death of D. bruxellensis during alcoholic fermentations.