Helmholtz Centre for Environmental Research - UFZ, Department of Proteomics, Leipzig, Germany.
J Appl Microbiol. 2010 Dec;109(6):1937-45. doi: 10.1111/j.1365-2672.2010.04823.x.
This study intended to unravel the physiological interplay in an anaerobic microbial community that degrades toluene under sulfate-reducing conditions combining proteomic and genetic techniques.
An enriched toluene-degrading community (Zz5-7) growing in batch cultures was investigated by DNA- and protein-based analyses. The affiliation and diversity of the community were analysed using 16S ribosomal RNA (rRNA) genes as a phylogenetic marker as well as bssA and dsrAB genes as functional markers. Metaproteome analysis was carried out by a global protein extraction and a subsequent protein separation by two-dimensional gel electrophoresis (2-DE). About 85% of the proteins in the spots were identified by nano-liquid chromatography coupled with electrospray mass spectrometry (nano-LC-ESI-MS/MS) analysis. DNA sequencing of bssA and the most abundant dsrAB amplicons revealed high similarities to a member of the Desulfobulbaceae, which was also predominant according to 16S rRNA gene amplicons. Metaproteome analysis provided 202 unambiguous protein identifications derived from 236 unique protein spots. The proteins involved in anaerobic toluene activation, dissimilatory sulfate reduction, hydrogen production/consumption and autotrophic carbon fixation were mainly affiliated to members of the Desulfobulbaceae and several other Deltaproteobacteria.
Phylogenetic and metaproteomic analyses revealed a member of the Desulfobulbaceae as the key player of anaerobic toluene degradation in a sulfate-reducing consortium.
This is the first study that combines genetic and proteomic analyses to indicate the interactions in an anaerobic toluene-degrading microbial consortium.
本研究旨在结合蛋白质组学和遗传技术，揭示在硫酸盐还原条件下降解甲苯的厌氧微生物群落中的生理相互作用。
通过 DNA 和蛋白质分析，研究了在批次培养中生长的富甲苯降解群落（Zz5-7）。使用 16S 核糖体 RNA（rRNA）基因作为系统发育标记以及 bssA 和 dsrAB 基因作为功能标记，分析了群落的归属和多样性。通过全局蛋白质提取和随后的二维凝胶电泳（2-DE）进行了元蛋白质组分析。斑点中约 85%的蛋白质通过纳升液相色谱-电喷雾质谱联用（nano-LC-ESI-MS/MS）分析进行鉴定。bssA 和最丰富的 dsrAB 扩增子的 DNA 测序揭示与脱硫杆菌科的一个成员高度相似，根据 16S rRNA 基因扩增子，该成员也是主要的。元蛋白质组分析提供了 202 个明确的蛋白质鉴定，源自 236 个独特的蛋白质斑点。涉及厌氧甲苯激活、异化硫酸盐还原、氢气产生/消耗和自养碳固定的蛋白质主要与脱硫杆菌科和其他几个 Delta 变形菌的成员有关。
系统发育和元蛋白质组学分析表明，脱硫杆菌科的一个成员是硫酸盐还原共生物群落中厌氧甲苯降解的关键参与者。
这是首次结合遗传和蛋白质组学分析，表明在厌氧甲苯降解微生物共生物群落中的相互作用。
