Fujian Provincial Key Laboratory of Agroecological Processing and Safety Monitoring, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, PR China; Key Laboratory of Crop Ecology and Molecular Physiology, Fujian Agriculture and Forestry University, Fujian Province University, Fuzhou 350002, PR China.
Department of Medical Laboratory, The 180th Hospital of PLA, Quanzhou, Fujian Province 362000, PR China.
J Proteomics. 2019 Sep 15;207:103468. doi: 10.1016/j.jprot.2019.103468. Epub 2019 Jul 30.
In this study, two susceptible strains and two multi-drug resistant clinical Escherichia coli strains were obtained by Kirby-Bauer method, and then a GC-MS-based metabolomics method was used to compare the differential expression of metabolites between two drug sensitive (CK1 and CK2) and two multidrug-resistant (MDR1 and MDR2) clinical strains of E. coli. We characterized a total of 273 metabolites, including 77 commonly altered metabolites, between MDR vs. antibiotic sensitive strains. Interestingly, the PCA score plot clearly discriminated drug sensitive and MDR strains. The following bioinformatics analysis showed that biosynthesis of amino acids, biosynthesis of phenylpropanoids and purine metabolism were commonly enriched in MDR strains. Moreover, microbial metabolism in diverse environments, carbon metabolism，and pyrimidine metabolism pathways were more likely to be enriched MDR1 strain, while ABC transporters, and cysteine and methionine metabolism pathways were enriched in MDR2 strains. The enzyme activities in several involved metabolic pathways were further measured and metabolite candidates were validated by GC-MS-SIM method. These results indicated that antibiotic resistance affects the metabolite profiles of bacteria. In general, our study provides evidence on the study and prediction of MDR characteristics and mechanisms in bacteria at the metabolite level. BIOLOGICAL SIGNIFICANCE: Overuse and abuse of antibiotics has led to the emergence of antibiotic-resistant strains of bacteria; however, relatively little is known about their resistance mechanisms. In this study, metabolomics method was used to compare the differential expression of metabolites between sensitive and multidrug-resistant clinical strains of E. coli. Results show that the PCA score plot clearly discriminated sensitive and MDR strains, indicating that they had different metabolic profiles. Further, bioinformatics analysis showed that biosynthesis of amino acids, biosynthesis of phenylpropanoids and purine metabolism may be related to resistance. Finally, the enzyme activities in several involved metabolic pathways were further measured and metabolite candidates were validated by GC-MS-SIM method. In general, our study provides evidence on the study and prediction of MDR characteristics and mechanisms in bacteria at the metabolite level.
在这项研究中，通过 Kirby-Bauer 法获得了两株敏感株和两株多药耐药临床大肠杆菌株，然后使用基于 GC-MS 的代谢组学方法比较了两株药敏（CK1 和 CK2）和两株多药耐药（MDR1 和 MDR2）临床大肠杆菌株之间代谢物的差异表达。我们共鉴定了 273 种代谢物，其中包括 MDR 与抗生素敏感株之间 77 种共同改变的代谢物。有趣的是，PCA 得分图清楚地区分了药敏和 MDR 菌株。以下生物信息学分析表明，氨基酸生物合成、苯丙素生物合成和嘌呤代谢在 MDR 菌株中普遍富集。此外，多样化环境中的微生物代谢、碳代谢和嘧啶代谢途径更可能在 MDR1 菌株中富集，而 ABC 转运蛋白、半胱氨酸和蛋氨酸代谢途径在 MDR2 菌株中富集。进一步测量了几个涉及代谢途径的酶活性，并通过 GC-MS-SIM 方法验证了代谢物候选物。这些结果表明，抗生素耐药性会影响细菌的代谢物谱。总的来说，我们的研究在代谢物水平上为细菌的 MDR 特性和机制的研究和预测提供了证据。生物意义：抗生素的过度和滥用导致了细菌抗生素耐药株的出现；然而，关于它们的耐药机制知之甚少。在这项研究中，使用代谢组学方法比较了敏感和多药耐药临床大肠杆菌株之间代谢物的差异表达。结果表明，PCA 得分图清楚地区分了敏感和 MDR 菌株，表明它们具有不同的代谢谱。进一步的生物信息学分析表明，氨基酸生物合成、苯丙素生物合成和嘌呤代谢可能与耐药性有关。最后，通过 GC-MS-SIM 方法进一步测量了几个涉及代谢途径的酶活性，并验证了代谢物候选物。总的来说，我们的研究在代谢物水平上为细菌的 MDR 特性和机制的研究和预测提供了证据。
