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Title: GAS CHROMATOGRAPHY-MASS SPECTROMETRY (GC-MS) TECHNIQUES FOR METABOLIC FLUX ANALYSIS OF THE BIFIDO SHUNT PATHWAY

Author
item Price, Neil
item Whitehead, Terence
item Cote, Gregory

Submitted to: Biocatalysis and Biotransformation
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/25/2005
Publication Date: 2/10/2006
Citation: Price, N.P., Whitehead, T.R., Cote, G.L. 2006. Global metabolic flux analysis of the bifidobacterial bifido shunt pathway. Biocatalysis and Biotransformation. 24(1-2):95-98.

Interpretive Summary: Bacteria of the family Bifidobacterium are beneficial in the human small intestine. These bacteria are said to be "probiotic," and may reduce or prevent disease. However, their enumeration in the gut is difficult, and the most direct tests involve certain tedious steps and can be prone to human error. In this paper, we propose an alternative approach to quantifying beneficial gut bacteria, by measuring rates of metabolic turnover of carbohydrate metabolites. The evaluation relies on sugar metabolism unique to these "good" bacteria. A more accurate method with less likelihood of human error would be of great advantage for evaluating the benefits of probiotic activity, both in humans and in livestock animals.

Technical Abstract: The presence of the Bifido shunt in Bifidobacterium is predicted to lead to the uptake and metabolic conversion of fructose to acetate. We propose an approach to quantifying the carbon flux through the Bifido shunt by measuring specific **13C-labeled carbohydrate-derived isotopomers by gas chromatography-mass spectrometry. The techniques described may provide an alternative approach for determining the in vitro prebiotic potential of dietary oligosaccharides and probiotic bacteria.