Proteomic investigation of amino acid catabolism in the indigenous gut anaerobe Fusobacterium varium

Authors

  • Joanna Potrykus,

    1. Department of Biochemistry and Molecular Biology, Dalhousie University, Halifax, Nova Scotia, Canada
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  • Robert L. White Dr.,

    1. Department of Chemistry, Dalhousie University, Halifax, Nova Scotia, Canada
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    • Additional corresponding author

  • Stephen L. Bearne Dr.

    Corresponding author
    1. Department of Biochemistry and Molecular Biology, Dalhousie University, Halifax, Nova Scotia, Canada
    2. Department of Chemistry, Dalhousie University, Halifax, Nova Scotia, Canada
    • Department of Biochemistry and Molecular Biology, Dalhousie University, Halifax, Nova Scotia, Canada B3H 1X5 Fax: +1-902-494-1355
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Abstract

The butyrate-producing anaerobe Fusobacterium varium is an integral constituent of human gut microflora. Unlike many gut microorganisms, F. varium is capable of fermenting both amino acids and glucose. Although F. varium has been implicated in beneficial as well as pathological bacterium–host interactions, its genome has not been sequenced. To obtain a better understanding of the metabolic processes associated with amino acid fermentation by F. varium, we used a gel-based proteomic approach to examine the changes in the soluble proteome accompanying the utilization of eight different growth substrates: glucose, L- and D-glutamate, L-histidine, L- and D-lysine, and L- and D-serine. Using LC-MS/MS to analyze ˜25% of the detected protein spots, we were able to identify 47 distinct proteins. While the intracellular concentrations of enzymes characteristic of a catabolic pathway for a specific amino acid were selectively increased in response to the presence of an excess of that amino acid in the growth medium, the concentrations of the core acetate–butyrate pathway enzymes remained relatively constant. Our analysis revealed (i) high intracellular concentrations of glutamate mutase and β-methylaspartate ammonia-lyase under all growth conditions, underscoring the importance of the methylaspartate pathway of glutamate catabolism in F. varium (ii) the presence of two enzymes of the hydroxyglutarate pathway of glutamate degradation in the proteome of F. varium ((R)-2-hydroxyglutaryl-CoA dehydratase and NAD-specific glutamate dehydrogenase) specifically when L-glutamate was the main energy source (iii) the presence of genes in the genome of F. varium encoding each of the enzymes of the hydroxyglutarate pathway (iv) the presence of both L- and D-serine ammonia-lyases (dehydratases) which permit F. varium to thrive on either L- or D-serine, respectively, and (v) the presence of aspartate-semialdehyde dehydrogenase and dihydrodipicolinate synthase, consistent with the ability of F. varium to synthesize meso-2,6-diaminopimelic acid as a component of its peptidoglycan. Proteins involved in other cellular processes, including oxidation–reduction reactions, protein synthesis and turnover, and transport were also identified.

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