4.5 Article

Extracellular metabolomics:: A metabolic footprinting approach to assess fiber degradation in complex media

Journal

ANALYTICAL BIOCHEMISTRY
Volume 349, Issue 2, Pages 297-305

Publisher

ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2005.11.019

Keywords

exometabolome; metabolite profiling; hydrolases; Clostridium proteoclasticum; Butyrivibrio fibrisolvens; Streptococcus bovis

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This work reports the implementation and optimization of a method for high-throughput analysis of metabolites produced by the breakdown of natural polysaccharides by microorganisms. Our simple protocol enables simultaneous separation and quantification of more than 40 different sugars and sugar derivatives, in addition to several organic acids in complex media, using 50-mu l samples and a standard gas chromatography mass spectrometry platform that was fully optimized for this purpose. As an implementation proof-of-concept, we assayed extracellular metabolite levels of three bacterial strains cultivated on complex medium rich in polysaccharides and under identical growth conditions. We demonstrate that the metabolic footprinting profile data distinguish among sample types such as typical metabolomics data. Moreover, we demonstrate that the differential metabolite-level data provide insight on specific fibrolytic activity of the different microbial strains and lay the groundwork for integrated proteome-metabolome studies of fiber-degrading microorganisms. (c) 2005 Elsevier Inc. All rights reserved.

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