Expression of coagulin A with low cytotoxic activity by Pediococcus pentosaceus ST65ACC isolated from raw milk cheese

Autor: Thomas Haertlé, Luís Augusto Nero, Svetoslav Dimitrov Todorov, Veselin Bivolarski, Anton Hinkov, Maria Ananieva, Stoyan Shishkov, Ilia Iliev, Daniel Todorov, Valeria Quintana Cavicchioli, Iskra Ivanova, Tonka Vasileva
Přispěvatelé: Faculdade de Ciências Farmacêuticas de Ribeirão Preto [São Paulo], Universidade de São Paulo (USP), Departamento de Veterinária, Universidade Federal de Vicosa (UFV), Department of Biochemistry and Microbiology, Faculty of Biology, Plovdiv University 'Paisii Hilendarski-Plovdiv University 'Paisii Hilendarski, Department of General and Applied Microbiology, Sofia University 'St. Kliment Ohridski'-Sofia University 'St. Kliment Ohridski', Laboratory of Virology, Unité de recherche sur les Biopolymères, Interactions Assemblages (BIA), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior Conselho Nacional de Desenvolvimento Cientifico e Tecnologico Fundacao de Amparo a Pesquisa do Estado de Minas Gerais
Rok vydání: 2019
Předmět:
Zdroj: Journal of Applied Microbiology
Journal of Applied Microbiology, Wiley, 2020, 128 (2), pp.458-472. ⟨10.1111/jam.14492⟩
ISSN: 1365-2672
1364-5072
Popis: Aims We aimed to evaluate some specific conditions for growth of Pediococcus pentosaceus ST65ACC and its bacteriocin expression through ABC transporters; to purify the bacteriocin and determine its sequence; and to evaluate the cytotoxicity potential of the purified bacteriocin(s). Methods and results The results presented for growth behaviour of P. pentosaceus ST65ACC showed that the bacterial growth was slightly influenced when cultured in MRS broth with different amounts of inoculum: 1, 2, 5 and 10%. The bacteriocin activity increased when 5 and 10% inocula were used. The carbon source (glucose) used in different amounts (1, 2, 3 or 4%) had no significant effect on growth and bacteriocin production. The studied strain P. pentosaceus ST65ACC was able to metabolize xylooligosaccharide (XOS) as the sole carbon source, resulting in the production of an antimicrobial peptide. The genes involved in the ABC transport system and sugar metabolism of P. pentosaceus ST65ACC were expressed at different levels. The bacteriocin produced by P. pentosaceus ST65ACC was partially purified by precipitation with ammonium sulphate (40% saturation), followed by reversed-phase liquid chromatography, resulting in the identification of an active bacteriocin. Tandem mass spectrometry was used to identify the partial sequence KYYGNGVTCGKHSCSVDWGK sharing high similarity to coagulin A. The semi-purified bacteriocin had low cytotoxicity based on estimated values for maximal nontoxic concentration (MNC) and cytotoxicity concentration (CC50 ). Conclusions The bacteriocin produced by P. pentosaceus ST65ACC is similar to coagulin, with low cytotoxicity, strong antimicrobial activity and possible additional metabolite routes in the producer cell. In addition to MRS broth, bacteriocin was produced also in medium containing XOS (as the single carbon source). Significance and impact of the study To the best of our knowledge, this is the first report of evaluation of the role of ABC transporters in the expression of bacteriocin by P. pentosaceus, cultured in MRS and XOS.
Databáze: OpenAIRE
Externí odkaz: