NDLI: Improving protein extraction and separation methods for investigating the metaproteome of anaerobic benzene communities within sediments

Content Provider	Springer Nature Link
Author	Benndorf, Dirk Vogt, Carsten Jehmlich, Nico Schmidt, Yvonne Thomas, Henrik Woffendin, Gary Shevchenko, Andrej Richw, Hans Hermann Bergen, Martin
Copyright Year	2009
Abstract	BTEX compounds such as benzene are frequent soil and groundwater contaminants that are easily biodegraded under oxic conditions by bacteria. In contrast, benzene is rather recalcitrant under anaerobic conditions. The analysis of anoxic degradation is often hampered by difficult sampling conditions, limited amounts of biomass and interference of matrix compounds with proteomic approaches. In order to improve the procedure for protein extraction we established a scheme consisting of the following steps: dissociation of cells from lava granules, cell lysis by ultrasonication and purification of proteins by phenol extraction. The 2D-gels revealed a resolution of about 240 proteins spots and the spot patterns showed strong matrix dependence, but still differences were detectable between the metaproteomes obtained after growth on benzene and benzoate. Using direct data base search as well as de novo sequencing approaches we were able to identify several proteins. An enoyl-CoA hydratase with cross species homology to Azoarcus evansii, is known to be involved in the anoxic degradation of xenobiotics. Thereby the identification confirmed that this procedure has the capacity to analyse the metaproteome of an anoxic living microbial community.
Starting Page	737
Ending Page	750
Page Count	14
File Format	PDF
ISSN	09239820
Journal	Biodegradation
Volume Number	20
Issue Number	6
e-ISSN	15729729
Language	English
Publisher	Springer Netherlands
Publisher Date	2009-04-21
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Extraction and purification of proteins Benzene biodegradation Microbial community Metaproteome analysis Contaminated groundwater Waste Management/Waste Technology Waste Water Technology / Water Pollution Control / Water Management / Aquatic Pollution Terrestrial Pollution Geochemistry Soil Science & Conservation Microbiology
Content Type	Text
Resource Type	Article
Subject	Environmental Chemistry Pollution Environmental Engineering Bioengineering Microbiology

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Improving protein extraction and separation methods for investigating the metaproteome of anaerobic benzene communities within sediments

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