Gryciuk Aleksander, Milner-Krawczyk Małgorzata, Rogalska Marta, Banach Artur Krzysztof, Sitkiewicz Ewa, Bakun Magdalena, Świadek Magdalena Edyta, Mierzejewska Jolanta
Microbiology and Bioengineering Laboratory, Faculty of Chemistry, Warsaw University of Technology, Warsaw, Poland.
MicroBioRobotic Systems Laboratory, Institute of Mechanical Engineering, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
Probiotics Antimicrob Proteins. 2025 Jun 13. doi: 10.1007/s12602-025-10624-0.
There is a huge disparity between the number of bacterial and yeast probiotics in favor of the former. The latest reports indicate that extracellular vehicles (EVs) play a significant role in probiotic mechanisms. In the present work, we compared the probiotic properties of Saccharomyces cerevisiae strains (WUT3 and WUT151), which have never been previously characterized in this context, with commercial probiotic yeast-Saccharomyces cerevisiae var. boulardii CNCM-745. Notably, WUT3 and WUT151 reacted more mildly to the unfavorable simulated environment of saliva, stomach, small, and large intestines. As a result, we confirmed that WUT3 and WUT151 were superior to S. boulardii in terms of probiotic properties. Then, we performed a complex analysis of their EVs, isolated by a multistep filtration process. The nanoparticle tracing analysis showed no significant difference in the diameter of the vesicles between the strains. MTT studies confirmed that EVs are not toxic against normal human colorectal cell lines CCD-18 Co and CCD 841 CoN. However, toxicity was observed against the HT-29 cancer line. By staining EVs with Nile Red, we successfully visualized EVs-cell interactions. Finally, we explored the profile of proteins transported with the EVs, identifying a significant overrepresentation of extracellular proteins. Based on comparison with other proteomic data, we selected marker proteins for S. cerevisiae EVs. This knowledge will be helpful for further studies on tracking the transfer of the protein cargo of yeast EVs to human cells using, for instance, specific antibodies to these marker proteins.
细菌益生菌和酵母益生菌的数量存在巨大差异，前者占优势。最新报告表明，细胞外囊泡（EVs）在益生菌机制中发挥着重要作用。在本研究中，我们将此前从未在此背景下进行过表征的酿酒酵母菌株（WUT3和WUT151）的益生菌特性与商业益生菌酵母——酿酒酵母变种布拉迪酵母CNCM-745进行了比较。值得注意的是，WUT3和WUT151对唾液、胃、小肠和大肠的不利模拟环境反应更为温和。结果，我们证实WUT3和WUT151在益生菌特性方面优于布拉迪酵母。然后，我们对通过多步过滤过程分离出的它们的细胞外囊泡进行了综合分析。纳米颗粒追踪分析表明，各菌株之间囊泡直径没有显著差异。MTT研究证实，细胞外囊泡对正常人结肠癌细胞系CCD-18 Co和CCD 841 CoN无毒。然而，观察到对HT-29癌细胞系有毒性。通过用尼罗红对细胞外囊泡进行染色，我们成功地观察到了细胞外囊泡与细胞的相互作用。最后，我们探索了随细胞外囊泡运输的蛋白质谱，确定细胞外蛋白质有显著的过度表达。基于与其他蛋白质组学数据的比较，我们选择了酿酒酵母细胞外囊泡的标记蛋白。这些知识将有助于进一步研究，例如使用针对这些标记蛋白的特异性抗体来追踪酵母细胞外囊泡的蛋白质货物向人类细胞的转移。
