Department of Microbiology, School of Natural Sciences, National University of Ireland, Galway, Ireland.
Electrophoresis. 2009 Dec;30(23):4149-51. doi: 10.1002/elps.200900474.
The feasibility of metaproteomic analysis of the microbial communities underpinning anaerobic wastewater treatment relies on efficient protein extraction and separation techniques. The microorganisms involved in anaerobic digestion (AD) of wastewater typically aggregate to form tightly organised, spherical granules, from which protein extraction is challenging. Here, we compare two methods of protein extraction, one using a French press previously used successfully to analyse the proteome of an activated sludge [Wilmes, P., Bond, P. L., Environ. Microbiol. 2004, 6, 911-920.] and one using sonication developed in the context of pure culture [Abram, F., Wan-Lin, S., Wiedmann, M., Boor, K. J., Coote, P., Botting, C., Karatzas, K. A. G., O'Byrne, C. P., Appl. Environ. Microbiol. 2008, 74, 594-604.]. We used 2-DE to carry out this comparison. The protein extraction using the sonication method resulted in a significant increase in the number of protein spots and higher quality 2-D gels.
基于微生物群落的厌氧废水处理的代谢蛋白质组分析的可行性依赖于有效的蛋白质提取和分离技术。参与废水厌氧消化（AD）的微生物通常聚集形成紧密组织的、球形的颗粒，从这些颗粒中提取蛋白质具有挑战性。在这里，我们比较了两种蛋白质提取方法，一种使用法国压榨机，该压榨机以前曾成功用于分析活性污泥的蛋白质组[Wilmes，P.，Bond，P. L.，Environ. Microbiol. 2004，6，911-920.]，另一种使用超声处理法，该方法是在纯培养的背景下开发的[Abram，F.，Wan-Lin，S.，Wiedmann，M.，Boor，K. J.，Coote，P.，Botting，C.，Karatzas，K. A. G.，O'Byrne，C. P.，Appl. Environ. Microbiol. 2008，74，594-604.]。我们使用 2-DE 进行了这一比较。使用超声处理法进行蛋白质提取导致蛋白质斑点数量显著增加，并且二维凝胶的质量更高。
