Revealing the insoluble metasecretome of lignocellulose-degrading microbial communities.
Revealing the insoluble metasecretome of lignocellulose-degrading microbial communities.
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DOI：
10.1038/s41598-017-02506-5
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发表时间：
2017-05-24
影响因子：
4.6
通讯作者：
Bruce NC
中科院分区：
文献类型：
--
作者：
Alessi AM;Bird SM;Bennett JP;Oates NC;Li Y;Dowle AA;Polikarpov I;Young JPW;McQueen-Mason SJ;Bruce NC
微生物群落利用分泌的酶代谢植物生物量；然而，在这些微生物组中鉴定与不溶性木质纤维素紧密结合的细胞外蛋白质是一个挑战，因为洗脱这些蛋白质所需的严格提取也会溶解与植物生物量相关的微生物，释放出污染后脑鞘的细胞内蛋白质。在这里，我们描述了一种针对细胞外蛋白质组的技术，该技术用于比较生长在小麦秸秆和水稻秸秆上的两种木质纤维素降解群落的元脑区和元表面蛋白质组。基于质谱的蛋白质组学与亚转录组学的结合能够从这些木质纤维素降解群落中鉴定出独特的分泌组池。该方法通过提高活性分泌蛋白和跨膜蛋白的检测，使我们能够有效地区分细胞外蛋白和细胞内蛋白。除了预期的碳水化合物活性酶外，我们的新方法还揭示了大量未知的蛋白质，支持了我们对微生物群落如何降解木质纤维素底物的理解存在重大差距的观点。
Microbial communities metabolize plant biomass using secreted enzymes; however, identifying extracellular proteins tightly bound to insoluble lignocellulose in these microbiomes presents a challenge, as the rigorous extraction required to elute these proteins also lyses the microbes associated with the plant biomass releasing intracellular proteins that contaminate the metasecretome. Here we describe a technique for targeting the extracellular proteome, which was used to compare the metasecretome and meta-surface-proteome of two lignocellulose-degrading communities grown on wheat straw and rice straw. A combination of mass spectrometry-based proteomics coupled with metatranscriptomics enabled the identification of a unique secretome pool from these lignocellulose-degrading communities. This method enabled us to efficiently discriminate the extracellular proteins from the intracellular proteins by improving detection of actively secreted and transmembrane proteins. In addition to the expected carbohydrate active enzymes, our new method reveals a large number of unknown proteins, supporting the notion that there are major gaps in our understanding of how microbial communities degrade lignocellulosic substrates.