Antibiotic-resistant bacteria are an increasingly serious threat to human health and aquaculture. To further explore bacterial antibiotic resistance mechanism, iTRAQ is used to identify a differential proteome in ampicillin-resistant LTB4 (LTB4-RAMP), a strain of Edwardsiella piscicida. A total of 102 differentially proteins with 50 upregulation and 52 downregulation are identified. Since many of these changes are related to metabolism, interactive pathways explorer(iPath) is used to understand a global differentially metabolic response in LTB4-RAMP. This analysis identifies a global depressed metabolic modulation as the most characteristic feature of LTB4-RAMP. Lower membrane potential and ATP in LTB4-RAMP than control support that the central carbon metabolism and energy metabolism are reduced. Since the pyruvate cycle (the P cycle) plays a key role in the central carbon metabolism and energy metabolism, further investigation focuses on the P cycle and shows that expression of genes and activity of enzymes in the P cycle are decreased in LTB4-RAMP. These results support the conclusion that the depressed P cycle contributes to the acquisition of ampicillin resistance in E.piscicida. These findings indicate that the combination of proteomics and iPath analysis can provide a global metabolic profile, which helps us better understand the correlation between ampicillin resistance and cellular metabolism. SIGNIFICANCE: The present study uses iTRAQ to explore ampicillin resistance mechanism in Edwardsiella piscicida and finds many of these differential abundances of proteins are related to metabolism. IPath further identifies a global depressed metabolic modulation and characterizes the reduced pyruvate cycle as the most characteristic feature of the ampicillin-resistant E. piscicida, which is supported by reduced expression of genes and activity of enzymes in the pyruvate cycle. Consisitently, lower membrane potential and ATP are detetced. These results reveal the metabolic mechanism of ampicillin resistance and provide a solid proof to revert the resistance by reprogramming metabolomics.

中文翻译：

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降低的P周期有助于在爱德华氏菌中获得氨苄青霉素抗性。

抗抗生素细菌对人类健康和水产养殖的威胁日益严重。为了进一步探索细菌对抗生素的耐药机制，iTRAQ用于鉴定耐氨苄青霉素的LTB4（LTB4-RAMP）（一种爱德华氏菌菌株）中的差异蛋白质组。总共鉴定出102种差异蛋白，其中50种上调和52种下调。由于这些变化中的许多变化都与代谢有关，因此使用交互式途径explorer（iPath）来了解LTB4-RAMP中的整体差异代谢反应。该分析将整体抑制的代谢调节确定为LTB4-RAMP的最典型特征。与对照相比，LTB4-RAMP中较低的膜电位和ATP支持降低中央碳代谢和能量代谢。由于丙酮酸循环（P循环）在中央碳代谢和能量代谢中起关键作用，因此进一步的研究集中在P循环上，并显示LTB4-RAMP中P循环中的基因表达和酶活性降低。这些结果支持这样的结论，即P周期的下降有助于大肠杆菌中氨苄青霉素抗性的获得。这些发现表明，蛋白质组学和iPath分析的结合可以提供整体代谢谱，​​这有助于我们更好地了解氨苄青霉素耐药性与细胞代谢之间的相关性。重要性：本研究使用iTRAQ来研究爱德华氏菌对氨苄青霉素的耐药机制，发现这些差异丰富的蛋白质中有许多与代谢有关。IPath进一步确定了整体抑制性的代谢调节，并将丙酮酸循环的降低表征为耐氨苄青霉素的大肠杆菌的最典型特征，丙酮酸循环中基因表达的降低和酶活性的降低也支持了这一点。因此，检测到较低的膜电位和ATP。这些结果揭示了氨苄青霉素抗性的代谢机制，并提供了通过重新编程代谢组学来逆转抗性的坚实证据。