Zorel J A, Aquino S F, Sanson A L, Castro-Borges W, Silva S Q
Núcleo de Pesquisa em Ciências Biológicas, Universidade Federal de Ouro Preto (UFOP), Brasil.
Departamento de Química, Universidade Federal de Ouro Preto (UFOP), Brasil.
Water Sci Technol. 2015;72(4):535-42. doi: 10.2166/wst.2015.244.
Despite the importance of anaerobic sludge extracellular polymeric substances (EPSs), their characterization is limited to information regarding their chemical classes and molecular size. This work explores the possibility of using proteomic techniques to study the proteins present in this matrix. Thus, this paper compares eight EPS extraction methods regarding extraction yield, protein/carbohydrate ratio, size distribution profile and suitability to sodium dodecyl sulfate-polyacrylamide gel electrophoresis analyses. Despite the differences found in quantification and size exclusion chromatography assays, the band profile found for all methods was very similar. Considering the band pattern, extraction time and background level, heating method followed by ammonium sulfate precipitation proved to be the most appropriate method for gel-based analyses of anaerobic sludge EPS proteins.
尽管厌氧污泥胞外聚合物（EPSs）很重要，但其表征仅限于有关其化学类别和分子大小的信息。本研究探索了使用蛋白质组学技术研究该基质中存在的蛋白质的可能性。因此，本文比较了八种EPS提取方法在提取产率、蛋白质/碳水化合物比、大小分布谱以及对十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析的适用性方面的差异。尽管在定量和尺寸排阻色谱分析中发现了差异，但所有方法的条带图谱非常相似。考虑到条带模式、提取时间和背景水平，加热法结合硫酸铵沉淀被证明是用于厌氧污泥EPS蛋白质基于凝胶分析的最合适方法。
