Currie Felicity, Broadhurst David I, Dunn Warwick B, Sellick Christopher A, Goodacre Royston
School of Chemistry, Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester, M1 7ND, UK.
Mol Biosyst. 2016 Apr;12(4):1367-77. doi: 10.1039/c5mb00889a. Epub 2016 Mar 2.
Human pharmaceuticals have been detected in wastewater treatment plants, rivers, and estuaries throughout Europe and the United States. It is widely acknowledged that there is insufficient information available to determine whether prolonged exposure to low levels of these substances is having an impact on the microbial ecology in such environments. In this study we attempt to measure the effects of exposing cultures of Pseudomonas putida KT2440 (UWC1) to six pharmaceuticals by looking at differences in metabolite levels. Initially, we used Fourier transform infrared (FT-IR) spectroscopy coupled with multivariate analysis to discriminate between cell cultures exposed to different pharmaceuticals. This suggested that on exposure to propranolol there were significant changes in the lipid complement of P. putida. Metabolic profiling with gas chromatography-mass spectrometry (GC-MS), coupled with univariate statistical analyses, was used to identify endogenous metabolites contributing to discrimination between cells exposed to the six drugs. This approach suggested that the energy reserves of exposed cells were being expended and was particularly evident on exposure to propranolol. Adenosine triphosphate (ATP) concentrations were raised in P. putida exposed to propranolol. Increased energy requirements may be due to energy dependent efflux pumps being used to remove propranolol from the cell.
在欧洲和美国各地的污水处理厂、河流及河口都检测到了人类使用的药物。人们普遍认为，目前尚无足够信息来确定长期接触低水平的这些物质是否会对这类环境中的微生物生态产生影响。在本研究中，我们试图通过观察代谢物水平的差异，来测定恶臭假单胞菌KT2440（UWC1）培养物暴露于六种药物后的影响。最初，我们使用傅里叶变换红外（FT-IR）光谱结合多变量分析，来区分暴露于不同药物的细胞培养物。这表明，恶臭假单胞菌在接触普萘洛尔后，其脂质成分发生了显著变化。利用气相色谱-质谱联用（GC-MS）进行代谢谱分析，并结合单变量统计分析，来识别有助于区分暴露于六种药物的细胞的内源性代谢物。这种方法表明，暴露细胞的能量储备正在消耗，在接触普萘洛尔时尤为明显。暴露于普萘洛尔的恶臭假单胞菌中三磷酸腺苷（ATP）浓度升高。能量需求增加可能是由于细胞使用了依赖能量的外排泵来清除普萘洛尔。
