Comparison of enzymatic activities and proteomic profiles of Butyrivibrio fibrisolvens grown on different carbon sources
Comparison of enzymatic activities and proteomic profiles of Butyrivibrio fibrisolvens grown on different carbon sources
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DOI：
10.1186/s12953-019-0150-3
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发表时间：
2019-06-01
期刊：
影响因子：
2
通讯作者：
Kopecny, Jan
中科院分区：
文献类型：
--
作者：
Sechovcova, Hana;Kulhava, Lucie;Kopecny, Jan
背景：瘤胃微生物区系是最复杂的厌氧菌群之一，涉及古生菌、细菌、原生动物、真菌和噬菌体。它们在利用植物多糖方面非常有效，特别是纤维素和半纤维素。最重要的半纤维素分解菌与丁核弧菌属聚在一起。由于相关菌种的水解活性和底物选择性不同，因此丁核弧菌被选为最有效的菌株之一，因此适合于胞外部分底物比较的蛋白质组学研究。纤维分离杆菌基因组是丁状弧菌中最大的，主要集中在“环境信息处理”和“碳水化合物代谢”上。方法：在木糖、葡萄糖、木聚糖、木聚糖和25%葡萄糖四种底物上培养，研究碳源对纤维分离杆菌3071的影响。用分光光度法和酶谱分析法对酶活性进行了研究。蛋白质组学研究基于基因组学、双向电泳法和NLC/MS(Bruker Daltonics)分析。结果：木聚糖可诱导胞外β-内切酶和木聚糖β-木糖苷酶活性。木聚糖聚合物的存在诱导了半纤维素酶的产生，并在40-80 kDa的范围内增加了蛋白质组分。纤维分离杆菌胞外3071蛋白的双向电泳和NLC/MS分析发现，14种不同的蛋白质在测试底物上的表达存在显著差异。结论：四种碳源的比较导致了纤维分离杆菌蛋白质组的主要显著变化发生在纤维溶解蛋白簇之外。受影响的蛋白质主要属于糖酵解和蛋白质合成簇。
Background: The rumen microbiota is one of the most complex consortia of anaerobes, involving archaea, bacteria, protozoa, fungi and phages. They are very effective at utilizing plant polysaccharides, especially cellulose and hemicelluloses. The most important hemicellulose decomposers are clustered with the genus Butyrivibrio. As the related species differ in their range of hydrolytic activities and substrate preferences, Butyrivibrio fibrisolvens was selected as one of the most effective isolates and thus suitable for proteomic studies on substrate comparisons in the extracellular fraction. The B. fibrisolvens genome is the biggest in the butyrivibria cluster and is focused on "environmental information processing" and "carbohydrate metabolism".Methods: The study of the effect of carbon source on B. fibrisolvens 3071 was based on cultures grown on four substrates: xylose, glucose, xylan, xylan with 25% glucose. The enzymatic activities were studied by spectrophotometric and zymogram methods. Proteomic study was based on genomics, 2D electrophoresis and nLC/MS (Bruker Daltonics) analysis.Results: Extracellular beta-endoxylanase as well as xylan beta-xylosidase activities were induced with xylan. The presence of the xylan polymer induced hemicellulolytic enzymes and increased the protein fraction in the interval from 40 to 80kDa. 2D electrophoresis with nLC/MS analysis of extracellular B. fibrisolvens 3071 proteins found 14 diverse proteins with significantly different expression on the tested substrates.Conclusion: The comparison of four carbon sources resulted in the main significant changes in B. fibrisolvens proteome occurring outside the fibrolytic cluster of proteins. The affected proteins mainly belonged to the glycolysis and protein synthesis cluster.