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

作为肺部感染模型的生物膜中铜绿假单胞菌代谢组的表征

• 批准号: 352241003
• 负责人: Professor Dr. Oliver Schmitz
• 金额: --
• 依托单位: Arbeitsgruppe Instrumentelle Analytische Chemie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2017
• 资助国家: 德国
• 起止时间: 2016-12-31 至 2020-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/352241003?language=en
• 关键词: Characterization; metabolome; P.; aeruginosa; biofilms

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.

在这个项目中，铜绿假单胞菌的代谢组将通过使用生物膜作为肺部感染的模型来表征肺适应条件。因此，将开发一种铜绿假单胞菌培养的采样系统，该系统能够随着时间的推移对培养基、生物膜和顶空进行采样。这种三阶段方法允许将检测到的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来实现代谢物的全面预浓缩。选定的代谢物将用GC-EI-MS和LC+LC-IM-qTOF-MS进行定性和定量测定。样品的预富集和实验参数，如固定相、二维分析时间(调制时间)和流速将被优化。将苯并噻唑作为内标添加到培养基中，以考察和控制潜在代谢物的预浓缩、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物来验证是否代表铜绿假单胞菌。这应该回答了是否只有通过顶空分析才能鉴定铜绿假单胞菌的问题。

项目成果

Analysis of volatile metabolites from in vitro biofilms of Pseudomonas aeruginosa with thin-film microextraction by thermal desorption gas chromatography-mass spectrometry

薄膜微萃取热解吸气相色谱-质谱法分析铜绿假单胞菌体外生物膜的挥发性代谢物

• DOI: 10.1007/s00216-020-02529-4
• 发表时间: 2020
• 期刊: Analytical and Bioanalytical Chemistry
• 影响因子: 4.3
• 作者: Koehler T;Ackermann I;Brecht D;Uteschil F;Wingender J;Telgheder U;Schmitz O. J.
• 通讯作者: Schmitz O. J.