Characterization of the metabolome of P. aeruginosa in biofilms as a lung infection model

在本项目中，将通过使用生物膜作为肺部感染模型，考虑肺部适应性条件，对铜绿假单胞菌的代谢组进行表征。因此，将开发用于铜绿假单胞菌培养的采样系统，其能够随时间对培养基、生物膜和顶空进行采样。这种三阶段方法允许将检测和表征的物质分类为底物或代谢物。应通过应用各种提取技术实现代谢物的综合预浓缩。将通过GCxGC-EI-MS和LC+LC-IM-qTOF-MS对选定代谢物进行定性和定量测定。将优化样品预浓缩和实验参数，例如固定相、二维分析时间（调制时间）和流速。 将苯并噻唑作为内标物加入培养基中，以研究和控制潜在代谢产物的预浓缩、扩散和分布。根据结果，应通过仅检测生物膜样品顶空中的代谢产物来验证铜绿假单胞菌是否代表。这将回答是否可以仅通过顶空分析来鉴定铜绿假单胞菌的问题。

In this project, the metabolome of P. aeruginosa will be characterized considering lung adapted conditions by using a biofilm as a lung infection model. Therefore, a sampling system for the cultivation of P. aeruginosa will be developed, which enables the sampling of the culture medium, the biofilm and the headspace over time. This three-phase approach allows the classification of detected and characterized substances as substrates or metabolites. A comprehensive preconcentration of the metabolites shall be achieved by application of various extraction techniques. The qualitative and quantitative determination of selected metabolites will be carried out by GCxGC-EI-MS and LC+LC-IM-qTOF-MS. The sample preconcentration and experimental parameters e.g. stationary phase, analysis time in the 2nd dimension (modulation time) and flow rates will be optimized. As an internal standard, Benzothiazole will be added to the culture medium in order to investigate and control the preconcentration, diffusion and distribution of the potential metabolites. Based on the results, it shall be verified if P. aeruginosa is represented by detecting only the metabolites in the headspace of a biofilm sample. This should answer the question whether the identification of P. aeruginosa is possible only by head space analysis or not.