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<p><i>Brevibacillus laterosporus</i> (<i>B. laterosporus</i>) is a significant probiotic bacterium known for producing multiple secondary metabolites with notable antibacterial activity. In this study, bioinformatic tools such as FastANI and Roary were employed to analyze genomic similarity and construct the <i>B. laterosporus</i> pan-genome. The software platforms antiSMASH and BAGEL4 were used to predict biosynthetic gene clusters (BGCs) and identify bacteriocins. Core candidate bacteriocins were subsequently subjected to heterologous expression in <i>E. coli</i>, followed by nickel column purification and concentration. Their antibacterial activity was systematically evaluated using agar diffusion assays, minimum inhibitory concentration (MIC) determinations, time-kill curves, scanning electron microscopy (SEM), and live/dead bacterial staining in <i>Listeria monocytogenes</i> (<i>L. monocytogenes</i>). Pan-genome analysis revealed an open genome for <i>B. laterosporus</i>, comprising 2,840 core genes. KEGG enrichment analyses indicated that these core genes were significantly associated with pathways responsible for the biosynthesis of antimicrobial compounds, including secondary metabolite, pan-quinone, and terpenoid quinone biosynthesis. AntiSMASH predicted 1,654 BGCs, predominantly non-ribosomal peptide synthase (NRPS), polyketide synthase (PKS), and heterocyclic amine-associated antimicrobial protein synthesis clusters. Moreover, BAGEL4 predicted three putative core candidate bacteriocin variants (BreL1, BreL2, and BreL3), all of which exhibited pronounced antibacterial activity against <i>L. monocytogenes</i>. Among these, BreL1 demonstrated the most robust activity, causing morphological changes consistent with membrane-associated damage. Collectively, this study elucidates the open pan-genome architecture and the metabolically rich secondary metabolome of <i>B. laterosporus</i>, identifies three putative bacteriocin variants with active antibacterial efficacy against <i>L. monocytogenes</i>, which provides a theoretical foundation for developing natural antimicrobial agents.</p> Graphical Abstract <p></p>

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Pan-genome guided identification and functional characterisation of three putative bacteriocin variants from Brevibacillus laterosporus

  • 期刊论文

  • 发表日期：

    2026年4月18日

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  • Ming Wu,
  • Fushuang Duan,
  • Shuang Chen,
  • Yuhang Nie,
  • Zhanpeng Wang,
  • Ying Yang,
  • Xuepeng Cai,
  • Jie Li,
  • Zhongmei Ma,
  • Qingling Meng,
  • Jun Qiao

摘要

Brevibacillus laterosporus (B. laterosporus) is a significant probiotic bacterium known for producing multiple secondary metabolites with notable antibacterial activity. In this study, bioinformatic tools such as FastANI and Roary were employed to analyze genomic similarity and construct the B. laterosporus pan-genome. The software platforms antiSMASH and BAGEL4 were used to predict biosynthetic gene clusters (BGCs) and identify bacteriocins. Core candidate bacteriocins were subsequently subjected to heterologous expression in E. coli, followed by nickel column purification and concentration. Their antibacterial activity was systematically evaluated using agar diffusion assays, minimum inhibitory concentration (MIC) determinations, time-kill curves, scanning electron microscopy (SEM), and live/dead bacterial staining in Listeria monocytogenes (L. monocytogenes). Pan-genome analysis revealed an open genome for B. laterosporus, comprising 2,840 core genes. KEGG enrichment analyses indicated that these core genes were significantly associated with pathways responsible for the biosynthesis of antimicrobial compounds, including secondary metabolite, pan-quinone, and terpenoid quinone biosynthesis. AntiSMASH predicted 1,654 BGCs, predominantly non-ribosomal peptide synthase (NRPS), polyketide synthase (PKS), and heterocyclic amine-associated antimicrobial protein synthesis clusters. Moreover, BAGEL4 predicted three putative core candidate bacteriocin variants (BreL1, BreL2, and BreL3), all of which exhibited pronounced antibacterial activity against L. monocytogenes. Among these, BreL1 demonstrated the most robust activity, causing morphological changes consistent with membrane-associated damage. Collectively, this study elucidates the open pan-genome architecture and the metabolically rich secondary metabolome of B. laterosporus, identifies three putative bacteriocin variants with active antibacterial efficacy against L. monocytogenes, which provides a theoretical foundation for developing natural antimicrobial agents.

Graphical Abstract