Extracellular metabolomics: a metabolic footprinting approach to assess fiber degradation in complex media |
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Authors: | Villas-Bôas Silas G Noel Samantha Lane Geoffrey A Attwood Graeme Cookson Adrian |
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Affiliation: | AgResearch Limited, Grasslands Research Centre, Private Bag 11008, Palmerston North 5301, New Zealand. silas.villas-boas@agresearch.co.nz |
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Abstract: | 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-mul 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. |
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Keywords: | Exometabolome Metabolite profiling Hydrolases Clostridium proteoclasticum Butyrivibrio fibrisolvens Streptococcus bovis |
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