Different culture media and purification methods unveil the core proteome of Propionibacterium freudenreichii -derived extracellular vesicles
2023
Rodovalho, Vinícius de Rezende | da Luz, Brenda Silva Rosa | Nicolas, Aurélie | Jardin, Julien | Briard-Bion, Valérie | Folador, Edson Luiz | Santos, Anderson Rodrigues | Jan, Gwénaël | Le Loir, Yves | de Carvalho Azevedo, Vasco Ariston | Guédon, Éric | Science et Technologie du Lait et de l'Oeuf (STLO) ; Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut Agro Rennes Angers ; Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro) | Universidade Federal de Minas Gerais = Federal University of Minas Gerais [Belo Horizonte, Brazil] (UFMG) | Universidade Estadual de Campinas = University of Campinas (UNICAMP) | Center of Biotechnology, Department of Biotechnology, Federal University of Paraíba, João Pessoa 58051-900 | Faculty of Computer Science, Department of Computer Science, Federal University of Uberlândia, Uberlândia 38400902 | a financial support from INRAE (Rennes, France) and Institut Agro (Rennes, France). Vinícius de Rezende Rodovalho and Brenda Silva Rosa da Luz were supported by the International Cooperation Program CAPES/COFECUBat the Federal University of Minas Gerais funded by CAPES—the Brazil- ian Federal Agency for the Support and Evaluation of Gradu-ate Education of the Brazilian Ministry of Education (number 99999.000058/2017-03 and 88887.179897/2018-00, respectively) | https://academic.oup.com/microlife/advance-article/doi/10.1093/femsml/uqad029/7189863?login=true
International audience
显示更多 [+] 显示较少 [-]英语. Bacterial extracellular vesicles (EVs) are natural lipidic nanoparticles implicated in intercellular communication. Although EV research focused mainly on pathogens, the interest in probiotic-derived EVs is now rising. One example is Propionibacterium freudenreichii, which produces EVs with anti-inflammatory effects on human epithelial cells. Our previous study with P. freudenreichii showed that EVs purified by size exclusion chromatography (SEC) displayed variations in protein content according to bacterial growth conditions. Considering these content variations, we hypothesized that a comparative proteomic analysis of EVs recovered in different conditions would elucidate whether a representative vesicular proteome existed, possibly providing a robust proteome dataset for further analysis. Therefore, P. freudenreichii was grown in two culture media, and EVs were purified by sucrose density gradient ultracentrifugation (UC). Microscopic and size characterization confirmed EV purification, while shotgun proteomics unveiled that they carried a diverse set of proteins. A comparative analysis of the protein content of UC- and SEC-derived EVs, isolated from cultures either in UF (cow milk ultrafiltrate medium) or YEL (laboratory yeast extract lactate medium), showed that EVs from all these conditions shared 308 proteins. This EV core proteome was notably enriched in proteins related to immunomodulation. Moreover, it showed distinctive features, including highly interacting proteins, compositional biases for some specific amino acids, and other biochemical parameters. Overall, this work broadens the toolset for the purification of P. freudenreichii-derived EVs, identifies a representative vesicular proteome, and enumerates conserved features in vesicular proteins. These results hold the potential for providing candidate biomarkers of purification quality, and insights into the mechanisms of EV biogenesis and cargo sorting.
显示更多 [+] 显示较少 [-]