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<p>Recently, more and more bacteria have been reported to become tolerant to antibiotics. In this study, one <i>tnaA</i> gene involved in indole production, and the effect of exogenous indole on the formation of persister cells specific to tetracycline in <i>Vibrio splendidus</i> were characterized. The <i>tnaA</i><sub><i>Vs</i></sub> gene was first cloned and conditionally expressed in <i>Escherichia coli</i> Rosetta (DE3). To investigate the regulatory effect of TnaA<sub>Vs</sub>, the <i>tnaA</i> deletion strain AJ01/Δ<i>tnaA</i> was constructed by in-frame deletion. The undetected extracellular indole in the AJ01/Δ<i>tnaA</i> indicated that TnaA was the solo enzyme to produce indole in <i>V. splendidus</i>. The drop plate method showed that AJ01/Δ<i>tnaA</i> was more tolerant to the higher concentration of tetracycline than that of AJ01, being 340-fold higher in the proportion of survived cells when cell density OD<sub>600</sub>≈0.65. Moreover, the synergistic effects of indole and tetracycline on killing of <i>V. splendidus</i> were determined. Results show that addition of 2-mmol/L indole increased the susceptibility of both AJ01 and AJ01/Δ<i>tnaA</i> to 10×minimum inhibitory concentration tetracycline. To explore the genes and pathways regulated by TnaA<sub>Vs</sub>, the transcriptomic analysis between AJ01 and AJ01/Δ<i>tnaA</i> was performed. Result shows that TCA cycle, arginine biosynthesis, quorum sensing and microbial metabolism in diverse environments were downregulated, while the ribosome pathways, the protein metabolic process, peptide biosynthetic and metabolic process were upregulated in the AJ01/Δ<i>tnaA</i>. This study shows that indole could enhance the bactericidal effect of tetracycline on <i>V. splendidus</i> by decreased ribosome level probably but increased ATP level.</p>

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Indole enhanced the bactericidal efficacy of tetracycline against pathogen Vibrio splendidus

  • 期刊论文

  • 发表日期：

    2025年4月10日

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  • Jinxia Zhang,
  • Zhixin Mu,
  • Qiling Zhou,
  • Weiwei Zhang

摘要

Recently, more and more bacteria have been reported to become tolerant to antibiotics. In this study, one tnaA gene involved in indole production, and the effect of exogenous indole on the formation of persister cells specific to tetracycline in Vibrio splendidus were characterized. The tnaAVs gene was first cloned and conditionally expressed in Escherichia coli Rosetta (DE3). To investigate the regulatory effect of TnaAVs, the tnaA deletion strain AJ01/ΔtnaA was constructed by in-frame deletion. The undetected extracellular indole in the AJ01/ΔtnaA indicated that TnaA was the solo enzyme to produce indole in V. splendidus. The drop plate method showed that AJ01/ΔtnaA was more tolerant to the higher concentration of tetracycline than that of AJ01, being 340-fold higher in the proportion of survived cells when cell density OD600≈0.65. Moreover, the synergistic effects of indole and tetracycline on killing of V. splendidus were determined. Results show that addition of 2-mmol/L indole increased the susceptibility of both AJ01 and AJ01/ΔtnaA to 10×minimum inhibitory concentration tetracycline. To explore the genes and pathways regulated by TnaAVs, the transcriptomic analysis between AJ01 and AJ01/ΔtnaA was performed. Result shows that TCA cycle, arginine biosynthesis, quorum sensing and microbial metabolism in diverse environments were downregulated, while the ribosome pathways, the protein metabolic process, peptide biosynthetic and metabolic process were upregulated in the AJ01/ΔtnaA. This study shows that indole could enhance the bactericidal effect of tetracycline on V. splendidus by decreased ribosome level probably but increased ATP level.