Elucidation of anaerobic benzene and alkylated aromatic compound degradation pathways and associated proteins by proteomic and metabolomic analyses

本项目主要研究苯和碱性芳香族化合物厌氧降解过程中的化学和生物影响以及对相关微生物的影响。在硫酸盐还原条件下苯降解的初始机制仍然未知，由于初步的蛋白质组学和宏基因组学实验仅提供了有限的线索，因此将对蛋白质和特定代谢物进行强化筛选。在蛋白质提取物的质谱分析期间，肽质量包含列表将允许优选检测感兴趣的肽。将通过质谱法对培养物进行总代谢组学筛选以及特异性CoA-酯鉴别。将在Protein-SIP实验中研究乙酸盐在群落代谢和烃底物特异性中的作用。在13 C标记的化合物上生长的培养物的代谢标记将允许检测代谢活性物质，因为该方法阐明了群落的功能和促生长信息并验证了检测到的代谢物。另一个重点将放在烷基化芳香族化合物在硫酸盐还原条件下的降解。由于初步研究表明，在间二甲苯降解培养物中存在与甲基萘降解蛋白相似的蛋白质，因此我们将通过蛋白质SIP实验和代谢物评估来研究该培养物。

The chemical and biological influences to anaerobic benzene and alkalyed aromatic compound degradation processes and to the involved microorganisms will be studied in this project. The initial mechanism in benzene degradation at sulfate-reducing conditions is still unknown and as preliminary proteomic and metagenomic experiments gave only limited clues an enhanced screening for proteins and specific metabolites will be carried out. During mass spectrometry of protein extracts peptide mass inclusion lists will allow a preferred detection of peptides of interest. A total metabolomic screening of the culture as well as specific CoA-Ester identifications will be done by mass spectrometry. The role of acetate in the community metabolism and the hydrocarbon substrate specificity will be investigated in Protein-SIP experiments. A metabolic labeling of the culture growing on 13C-labeled compounds will allow the detection of metabolic active species as the method elucidates the functional and phylogenic information of the community and validates the metabolites detected. Another focus will be laid on the alkylated aromatic compound degradation under sulfate reducing conditions. As preliminary investigations showed the presence of proteins similar to methylnaphthalene degrading proteins in a m-xylene degrading culture we will study the culture by Protein-SIP experiments and metabolite assessment.