Prediction of RNAs in <i>P</i>. <i>pentosaceus</i> MBBL6.

<div><p><i>Pediococcus pentosaceus</i> is well known for its probiotic properties, including roles in improving health, antimicrobial production, and enhancing fermented food quality. This study aimed to comprehensively analyze the whole genome of <i>P</i>. <i>pentosaceus</i> MBBL6, isolated from healthy cow milk, to assess its probiotic and antimicrobial potentials. <i>P</i>. <i>pentosaceus</i> MBBL6, isolated from a healthy cow milk at BSMRAU dairy farm, Gazipur, Bangladesh, underwent comprehensive genomic analysis, including whole genome sequencing, assembly, annotation, phylogenetic comparison, and assessment of metabolic pathways and secondary metabolites. Antimicrobial efficacy was evaluated through <i>in-vitro</i> and <i>in-vivo</i> studies, alongside <i>in-silico</i> exploration for potential mastitis therapy. We predicted 1,906 genes and 204 SEED sub-systems involved in carbohydrate metabolism and vitamin B complex biosynthesis, with a focus on lactose metabolism in MMBL6. Notably, 43 putative carbohydrate-active enzyme genes, including lysozymes, suggest the ability of MBBL6 for carbohydrate biotransformation and antimicrobial activity. The genome also revealed primary metabolic pathways for arginine and gallic acid metabolism and secondary metabolite gene clusters, including T3PKS and RiPP-like regions. Importantly, two bacteriocin biosynthesis gene clusters namely <i>bovicin_255</i>_variant and <i>penocin_A</i>, were identified in MBBL6. The safety assessment of MBBL6 genome revealed no virulence genes and a low pathogenicity score (0.196 out of 1.0). Several genes related to survival in gastrointestinal tract and colonization were also identified. Furthermore, MBBL6 exhibited susceptibility to a wide range of antibiotics <i>in-vitro</i>, and effectively suppressed mastitis pathogens in an <i>in-vivo</i> mouse mastitis model trial. The observed bacteriocin, particularly bovicin, demonstrated the ability to disrupt the function of an essential protein, Rho factor of mastitis pathogens by blocking transcription termination process. Taken together, our in-depth genomic analysis underscores the metabolic versatility, safety profile, and antimicrobial potential of <i>P</i>. <i>pentosaceus</i> MBBL6, suggesting its promise for applications in therapeutics, bioremediation, and biopreservation.</p></div>