National Research Laboratory of Molecular Microbiology and Toxicology, Department of Agricultural Biotechnology, Seoul National University, Seoul 08826, Republic of Korea.
Center for Food and Bioconvergence, Seoul National University, Seoul 08826, Republic of Korea.
J Microbiol Biotechnol. 2024 Nov 28;34(11):2201-2210. doi: 10.4014/jmb.2406.06052. Epub 2024 Sep 20.
, an opportunistic human pathogen, employs biofilm formation as a key survival and virulence mechanism. BrpT, a transcriptional regulator, is essential for biofilm development by regulating the expression of biofilm-related genes. In this study, we aimed to identify a small molecule inhibitor of BrpT to combat biofilm formation. High-throughput screening of 7,251 compounds using an reporter strain carrying the arabinose-inducible gene and a BrpT-activated promoter fused to the operon identified a hit compound, BTI (BrpT Inhibitor). BTI potently inhibited BrpT activity in (EC of 6.48 μM) without affecting bacterial growth or host cell viability. Treatment with BTI significantly reduced the expression of the BrpT regulon and impaired biofilm formation and colony rugosity in , thus increasing its susceptibility to antibiotics. In vitro biochemical analyses revealed that BTI directly binds to BrpT and inhibits its transcriptional regulatory activity. The identification of BTI as a specific inhibitor of BrpT that effectively diminishes biofilm formation provides a promising foundation for the development of novel anti-biofilm strategies, with the potential to address the growing challenge of antibiotic resistance and improve the treatment of biofilm-associated infections.
，一种机会性病原体，采用生物膜形成作为关键的生存和毒力机制。BrpT 是一种转录调节因子，通过调节生物膜相关基因的表达对 生物膜的发育是必不可少的。在这项研究中，我们旨在鉴定 BrpT 的小分子抑制剂以对抗 生物膜的形成。使用携带阿拉伯糖诱导的 基因和与 BrpT 激活启动子融合的 操纵子的报告菌株对 7251 种化合物进行高通量筛选，鉴定出一种有效化合物 BTI（BrpT 抑制剂）。BTI 强烈抑制 中的 BrpT 活性（EC 的 6.48 μM），而不影响细菌生长或宿主细胞活力。用 BTI 处理可显著降低 BrpT 调控子的表达，并损害 生物膜的形成和菌落粗糙度，从而增加其对抗生素的敏感性。体外生化分析表明 BTI 直接与 BrpT 结合并抑制其转录调节活性。鉴定 BTI 作为 BrpT 的特异性抑制剂，可有效减少 生物膜的形成，为开发新型抗生物膜策略提供了有希望的基础，有潜力解决日益严重的抗生素耐药性问题并改善生物膜相关感染的治疗效果。
