Optimisation of protein extraction and 2-DE for metaproteomics of microbial communities from anaerobic wastewater treatment biofilms
Optimisation of protein extraction and 2-DE for metaproteomics of microbial communities from anaerobic wastewater treatment biofilms
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DOI：
10.1002/elps.200900474
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发表时间：
2009-12-01
期刊：
影响因子：
2.9
通讯作者：
O'Flaherty, Vincent
中科院分区：
文献类型：
--
作者：
Abram, Florence;Gunnigle, Eoin;O'Flaherty, Vincent
对支撑厌氧废水处理的微生物群落进行元蛋白质组学分析的可行性依赖于高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颗粒，从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，一种是使用先前成功用于分析活性污泥的蛋白质组的法国压榨机[Wilmes，P.，邦德，PL，Environ. Microbiol. 2004，6，911-920]和一种使用在纯培养物的背景下开发的超声处理[Abram，F.，Wan-Lin，S.，Wiedmann，M，Boor，K J.，Coote，P.，博廷角，卡拉扎斯湾A. G.，奥伯恩角P，Appl. Environ. Microbiol. 2008，74，594-604.]。我们使用2-DE进行比较。使用超声法的蛋白质提取导致蛋白质点的数量和更高质量的2-D凝胶的显着增加。
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.