McAnulty Michael J, Guron Giselle K, Oest Adam M, Miller Amanda L, Renye John A
Dairy and Functional Foods Research Unit, Eastern Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, Wyndmoor, PA, United States.
Front Microbiol. 2024 Jan 8;14:1304136. doi: 10.3389/fmicb.2023.1304136. eCollection 2023.
Bacteriocin production in is regulated by cell density-dependent signaling molecules, including BlpC, which regulates transcription from within the bacteriocin-like peptide () gene cluster. In some strains, such as ST106, this signaling system does not function properly, and BlpC must be supplied exogenously to induce bacteriocin production. In other strains, such as B59671, bacteriocin (thermophilin 110 in strain B59671) production occurs naturally. Here, transcriptomic analyses were used to compare global gene expression within ST106 in the presence or absence of synthetic BlpC and within B59671 to determine if BlpC regulates the expression of genes outside the cluster. Real-time semi-quantitative PCR was used to find genes differentially expressed in the absence of chromosomal in the B59671 background. Growth curve experiments and bacteriocin activity assays were performed with knockout mutants and BlpC supplementation to identify effects on growth and bacteriocin production. In addition to the genes involved in bacteriocin production, BlpC affected the expression of several transcription regulators outside the gene cluster, including a putative YtrA-subfamily transcriptional repressor. In strain B59671, BlpC not only regulated the expression of thermophilin 110 but also suppressed the production of another bacteriocin, thermophilin 13, and induced the same YtrA-subfamily transcriptional repressor identified in ST106. Additionally, it was shown that the broad-spectrum antimicrobial activity associated with strain B59671 was due to the production of thermophilin 110, while thermophilin 13 appears to be a redundant system for suppressing intraspecies growth. BlpC production or induction negatively affected the growth of strains B59671 and ST106, revealing selective pressure to not produce bacteriocins that may explain bacteriocin production phenotype differences between strains. This study identifies additional genes regulated by BlpC and assists in defining conditions to optimize the production of bacteriocins for applications in agriculture or human and animal health.
细菌素的产生在[具体细菌名称未给出]中受细胞密度依赖性信号分子调控,包括BlpC,它调控类细菌素样肽([具体肽名称未给出])基因簇内的转录。在某些菌株中,如[具体细菌名称未给出] ST106,这种信号系统功能不正常,必须外源提供BlpC以诱导细菌素产生。在其他菌株中,如[具体细菌名称未给出] B59671,细菌素(B59671菌株中的嗜热菌素110)自然产生。在此,转录组分析用于比较在有或无合成BlpC情况下ST106内的全局基因表达以及B59671内的全局基因表达,以确定BlpC是否调控[具体基因簇名称未给出]基因簇外基因的表达。实时半定量PCR用于在B59671背景下寻找在无染色体[具体基因名称未给出]时差异表达的基因。用敲除突变体和补充BlpC进行生长曲线实验和细菌素活性测定,以确定对生长和细菌素产生的影响。除了参与细菌素产生的基因外,BlpC还影响[具体基因簇名称未给出]基因簇外几个转录调节因子的表达,包括一个假定的YtrA亚家族转录抑制因子。在B59671菌株中,BlpC不仅调控嗜热菌素110的表达,还抑制另一种细菌素嗜热菌素13的产生,并诱导在ST106中鉴定出的相同YtrA亚家族转录抑制因子。此外,研究表明与B59671菌株相关的广谱抗菌活性归因于嗜热菌素110的产生,而嗜热菌素13似乎是抑制种内生长的冗余系统。BlpC的产生或诱导对B59671和ST106菌株的生长有负面影响,揭示了不产生细菌素的选择压力,这可能解释了[具体细菌名称未给出]菌株间细菌素产生表型差异。本研究鉴定了受BlpC调控的其他基因,并有助于确定优化细菌素生产以用于农业或人类及动物健康应用的条件。