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Article

Isolation, Genomics-Based and Biochemical Characterization of Bacteriocinogenic Bacteria and Their Bacteriocins, Sourced from the Gastrointestinal Tract of Meat-Producing Pigs

by
Ester Sevillano
,
Irene Lafuente
,
Nuria Peña
,
Luis M. Cintas
,
Estefanía Muñoz-Atienza
*,
Pablo E. Hernández
and
Juan Borrero
Departamento de Nutrición y Ciencia de los Alimentos (NUTRYCIAL), Sección Departamental de Nutrición y Ciencia de los Alimentos (SD-NUTRYCIAL), Facultad de Veterinaria, Universidad Complutense de Madrid (UCM), Avenida Puerta de Hierro, s/n, 28040 Madrid, Spain
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2024, 25(22), 12210; https://doi.org/10.3390/ijms252212210
Submission received: 21 October 2024 / Revised: 8 November 2024 / Accepted: 12 November 2024 / Published: 14 November 2024
(This article belongs to the Special Issue Applications of Multi-omics Technologies to Comprehend Properties and Functions of Probiotics)
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Abstract

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.
Keywords: bacteriocin-producing bacteria; bacteriocins; AMR; whole genome sequencing (WGS); bacteriocin gene clusters (BGC); in vitro cell-free protein synthesis (IV-CFPS); IV-CFPS/SIML; MALDI-TOF MS; LC-MS-MS bacteriocin-producing bacteria; bacteriocins; AMR; whole genome sequencing (WGS); bacteriocin gene clusters (BGC); in vitro cell-free protein synthesis (IV-CFPS); IV-CFPS/SIML; MALDI-TOF MS; LC-MS-MS

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MDPI and ACS Style

Sevillano, E.; Lafuente, I.; Peña, N.; Cintas, L.M.; Muñoz-Atienza, E.; Hernández, P.E.; Borrero, J. Isolation, Genomics-Based and Biochemical Characterization of Bacteriocinogenic Bacteria and Their Bacteriocins, Sourced from the Gastrointestinal Tract of Meat-Producing Pigs. Int. J. Mol. Sci. 2024, 25, 12210. https://doi.org/10.3390/ijms252212210

AMA Style

Sevillano E, Lafuente I, Peña N, Cintas LM, Muñoz-Atienza E, Hernández PE, Borrero J. Isolation, Genomics-Based and Biochemical Characterization of Bacteriocinogenic Bacteria and Their Bacteriocins, Sourced from the Gastrointestinal Tract of Meat-Producing Pigs. International Journal of Molecular Sciences. 2024; 25(22):12210. https://doi.org/10.3390/ijms252212210

Chicago/Turabian Style

Sevillano, Ester, Irene Lafuente, Nuria Peña, Luis M. Cintas, Estefanía Muñoz-Atienza, Pablo E. Hernández, and Juan Borrero. 2024. "Isolation, Genomics-Based and Biochemical Characterization of Bacteriocinogenic Bacteria and Their Bacteriocins, Sourced from the Gastrointestinal Tract of Meat-Producing Pigs" International Journal of Molecular Sciences 25, no. 22: 12210. https://doi.org/10.3390/ijms252212210

APA Style

Sevillano, E., Lafuente, I., Peña, N., Cintas, L. M., Muñoz-Atienza, E., Hernández, P. E., & Borrero, J. (2024). Isolation, Genomics-Based and Biochemical Characterization of Bacteriocinogenic Bacteria and Their Bacteriocins, Sourced from the Gastrointestinal Tract of Meat-Producing Pigs. International Journal of Molecular Sciences, 25(22), 12210. https://doi.org/10.3390/ijms252212210

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