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Optimisation of protein extraction and 2‐DE for metaproteomics of microbial communities from anaerobic wastewater treatment biofilms
Author(s) -
Abram Florence,
Gunnigle Eoin,
O'Flaherty Vincent
Publication year - 2009
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200900474
Subject(s) - metaproteomics , sonication , extraction (chemistry) , context (archaeology) , wastewater , anaerobic exercise , chromatography , protein purification , anaerobic digestion , activated sludge , chemistry , proteome , microbiology and biotechnology , biology , proteomics , biochemistry , environmental science , environmental engineering , gene , physiology , paleontology , organic chemistry , methane
Abstract 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.