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Metaproteomics of anaerobic microbial communities degrading long-chain fatty acids

机译:厌氧微生物群落降解长链脂肪酸的蛋白质组学

摘要

The anaerobic conversion of long-chain fatty acids (LCFA), and specifically the difference between the degradation of unsaturated- and saturated-LCFA, is not fully understood. In this work, syntrophic degradation of stearate (C18:0) and oleate (C18:1) was studied. A comparative metaproteomics approach, in which proteins were analyzed by LC-MS/MS, was combined with 16S rRNA gene pyrosequencing. Saturated- and unsaturated-LCFA were converted to methane by the anaerobic consortia. 16S rRNA gene pyrosequencing revealed differences in the microbial composition of sludges incubated with stearate and oleate, separately. Abundance of microorganisms within Deltaproteobacteria and within Synergistia taxa was higher in stearate and oleate incubations, respectively. Methanosaeta was the most abundant methanogen in both conditions. Metaproteomics results were similar and comparable distributions of COG functional categories were found for both samples. Archaeal proteomes were much better identified than bacterial ones, with five times more proteins retrieved. Most of the proteins identified belong to Methanosaeta concilli and Syntrophobacter fumaroxidans, two organisms that have their genome sequenced. Syntrophobacter belongs to Deltaproteobacteria, however this group was not dominant in oleate incubation as determined by pyrosequencing results. Studying metaproteomes of complex microbial communities is still a big challenge especially because most of the genomes are not sequenced which hinders protein identification.
机译:长链脂肪酸(LCFA)的厌氧转化,特别是不饱和和饱和LCFA降解之间的差异,尚未完全了解。在这项工作中,研究了硬脂酸酯(C18:0)和油酸酯(C18:1)的同养降解。一种比较元蛋白质组学方法,其中通过LC-MS / MS分析蛋白质,与16S rRNA基因焦磷酸测序结合。厌氧财团将饱和LCFA和不饱和LCFA转化为甲烷。 16S rRNA基因焦磷酸测序揭示了分别与硬脂酸酯和油酸酯孵育的污泥微生物组成的差异。在硬脂酸盐和油酸盐培养中,Deltaproteobacteria和Synergistia类群内的微生物丰富度分别较高。在两种情况下,甲烷菌属都是最丰富的产甲烷菌。元蛋白质组学结果相似,并且两种样品的COG功能类别分布均相当。与细菌相比,古细菌蛋白质组的鉴定要好得多,蛋白质的回收率是细菌的五倍。鉴定出的大多数蛋白质都属于康斯坦甲烷单胞菌(Methanosaeta concilli)和富氧氧化梭菌(Syntrophobacter fumaroxidans),这两种生物已对其基因组测序。滑膜细菌属于Deltaproteobacteria,但是根据焦磷酸测序结果,该组在油酸培养中并不占优势。研究复杂的微生物群落的元蛋白质组仍然是一个巨大的挑战,特别是因为大多数基因组未测序,这阻碍了蛋白质的鉴定。

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