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

本课题将对厌氧苯和烷基芳香族化合物的降解过程以及相关微生物的化学和生物影响进行研究。硫酸盐还原条件下苯降解的初步机制尚不清楚，由于初步的蛋白质组和元基因组学实验只给出了有限的线索，将对蛋白质和特定代谢物进行加强筛选。在蛋白质提取物的质谱分析过程中，多肽质量包含物列表将允许优先检测感兴趣的多肽。培养物的全部代谢组筛选以及特定的CoA-Ester鉴定将通过质谱学进行。醋酸酯在群落代谢中的作用和碳氢化合物底物的专一性将在蛋白质-SIP实验中进行研究。对生长在13C标记化合物上的培养物进行代谢标记将允许检测代谢活性物种，因为该方法阐明了群落的功能和系统发育信息，并验证了检测到的代谢物。另一个重点将放在硫酸盐还原条件下烷基芳香族化合物的降解。由于初步研究表明，在间二甲苯降解培养物中存在类似于甲基萘降解蛋白的蛋白质，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.