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<p>The persistence of heat-resistant <i>Bacillus subtilis</i> spores in dairy products constitutes a critical constraint on product quality and safety. Currently, thermosonication technology demonstrates considerable potential as a novel non-thermal sterilization method for spore inhibition. However, its precise mechanism of action remains unclear. To address this research gap, this study employed an integrated transcriptomic and proteomic approach to systematically investigate the inhibitory mechanism of thermosonication against <i>Bacillus subtilis</i> SN-6. Transcriptomic analysis revealed significant downregulation of multiple spore coat-associated proteins following thermosonication treatment, including six YjcZ family sporulation proteins, YqfC, and SpoVM. Proteomics results revealed that the expression of 13 <i>α</i>/<i>β</i>-type small acid-soluble spore proteins (SASPs), the GerD germination receptor, the SleB enzyme, and the activator protein (RBAM_001820) in the KinB signaling pathway was significantly downregulated. Integrated multi-omics analysis demonstrated that thermosonication exerted multi-level inhibitory effects through regulation of spore-related genes, impacting spore coat formation, spore protein composition, germination capacity, and signal transduction. Furthermore, it also exerts an indirect inhibitory effect on spore by suppressing key metabolic pathways, including uridine nucleotide synthesis, arginine and proline metabolism, and TCA cycle. Finally, RT-qPCR was used to validate the two key genes identified through a multi-omics approach. This study systematically elucidates the dual inhibitory mechanisms of thermosonication against <i>Bacillus subtilis</i> spores through integrated transcriptomic-proteomic analysis, identifying critical genes at the regulatory level and clarifying indirect suppression through metabolic network disruption. These findings provide a theoretical foundation for developing novel control strategies against heat-resistant spores and support the future application of thermosonication technology in the dairy industry.</p>

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Mechanism of Thermosonication in Inhibiting Spore of Heat-Resistant Bacillus subtilis via Metabolic Networks and Gene Regulation

  • 期刊论文

  • 发表日期：

    2026年7月27日

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  • Lisha Song,
  • Kairu He,
  • Wang Han,
  • Yuxin Zhang,
  • Yufei Bao,
  • Jie Min,
  • Shuangyu Miao,
  • Fangjian Cheng,
  • Rina Wu,
  • Zhishen Mu,
  • Junrui Wu

摘要

The persistence of heat-resistant Bacillus subtilis spores in dairy products constitutes a critical constraint on product quality and safety. Currently, thermosonication technology demonstrates considerable potential as a novel non-thermal sterilization method for spore inhibition. However, its precise mechanism of action remains unclear. To address this research gap, this study employed an integrated transcriptomic and proteomic approach to systematically investigate the inhibitory mechanism of thermosonication against Bacillus subtilis SN-6. Transcriptomic analysis revealed significant downregulation of multiple spore coat-associated proteins following thermosonication treatment, including six YjcZ family sporulation proteins, YqfC, and SpoVM. Proteomics results revealed that the expression of 13 α/β-type small acid-soluble spore proteins (SASPs), the GerD germination receptor, the SleB enzyme, and the activator protein (RBAM_001820) in the KinB signaling pathway was significantly downregulated. Integrated multi-omics analysis demonstrated that thermosonication exerted multi-level inhibitory effects through regulation of spore-related genes, impacting spore coat formation, spore protein composition, germination capacity, and signal transduction. Furthermore, it also exerts an indirect inhibitory effect on spore by suppressing key metabolic pathways, including uridine nucleotide synthesis, arginine and proline metabolism, and TCA cycle. Finally, RT-qPCR was used to validate the two key genes identified through a multi-omics approach. This study systematically elucidates the dual inhibitory mechanisms of thermosonication against Bacillus subtilis spores through integrated transcriptomic-proteomic analysis, identifying critical genes at the regulatory level and clarifying indirect suppression through metabolic network disruption. These findings provide a theoretical foundation for developing novel control strategies against heat-resistant spores and support the future application of thermosonication technology in the dairy industry.