RT Journal Article
T1 Isolation, Genomics-Based and Biochemical Characterization of Bacteriocinogenic Bacteria and Their Bacteriocins, Sourced from the Gastrointestinal Tract of Meat-Producing Pigs
A1 Sevillano, Ester
A1 Peña Vidal, Nuria
A1 Cintas Izarra, Luis Miguel
A1 Muñoz Atienza, Estefanía
A1 Hernández Cruza, Pablo Elpidio
A1 Borrero Del Pino, Juan
AB Antimicrobial resistance (AMR) poses a significant challenge to animal production due to the widespread use of antibiotics. Therefore, there is an urgent need for alternative antimicrobial strategies to effectively manage bacterial infections, protect animal health, and reduce reliance on antibiotics. This study evaluated the use of emerging approaches and procedures for the isolation, identification, and characterization of bacteriocin-producing bacteria and their bacteriocins, sourced from the gastrointestinal tract (GIT) of meat-producing pigs. Out of 2056 isolates screened against Gram-positive and Gram-negative indicator strains, 20 of the most active antimicrobial isolates were subjected to whole genome sequencing (WGS) for the prediction of coding DNA sequences (CDS) and the identification of bacteriocin gene clusters (BGC) and their functions. The use of an in vitro cell-free protein synthesis (IV-CFPS) protocol and the design of an IV-CFPS coupled to a split-intein mediated ligation (IV-CFPS/SIML) procedure made possible the evaluation of the production and antimicrobial activity of described and putatively novel bacteriocins. A colony MALDI-TOF MS procedure assisted in the identification of class I, II, and III lanthipeptides. MALDI-TOF MS and a targeted proteomics, combined with a massive peptide analysis (LC-MS/MS) approach, has proven valuable for the identification and biochemical characterization of previously described and novel bacteriocins encoded by the isolated bacteriocin-producing strains.
PB MDPI
SN 1661-6596
YR 2024
FD 2024
LK https://hdl.handle.net/20.500.14352/112359
UL https://hdl.handle.net/20.500.14352/112359
LA eng
NO Sevillano, E.; Lafuente, I.; Peña, N.; Cintas, L.M.; MuñozAtienza, E.; Hernández, P.E.; Borrero, J. Isolation, Genomics-Based and Biochemical Characterization of Bacteriocinogenic Bacteria and Their Bacteriocins, Sourced from the Gastrointestinal Tract of MeatProducing Pigs. Int. J. Mol. Sci. 2024, 25, 12210. https://doi.org/10.3390/ ijms252212210
NO Author Contributions: Conceptualization, E.M.-A., P.E.H. and J.B.; methodology, E.S., N.P., I.L., P.E.H. and J.B.; investigation, E.S., N.P., I.L., E.M.-A. and J.B.; resources, L.M.C., E.M.-A., P.E.H. andJ.B.; data curation, E.S., P.E.H. and J.B.; writing—original draft preparation, E.S.; writing—review and editing, P.E.H. and J.B.; supervision, E.M.-A., P.E.H. and J.B.; project administration, J.B.; fundingacquisition, L.M.C., P.E.H. and J.B. All authors have read and agreed to the published version of the manuscript.Acknowledgments: The authors also thank the Proteomics Unit of the University Complutense of Madrid (UCM), integrated ProteoRed, PRB3-ISCIII, and support by grant PT17/0019 of the PEI+D+I 2013–2016, funded by ISCIII and ERDF, for the technical assistance in the proteomic analysis of samples. This work conforms the Nagoya Protocol ESNC99 (ABSCH-IRCC-ES-257657-1).
NO Ministerio de Ciencia, Innovación y Universidades (España)
NO Comunidad de Madrid
DS Docta Complutense
RD 22 abr 2025