NMR Detection of ¹³CH₃¹³COOH from 3-¹³C-Glucose: A Signature for Bifidobacterium Fermentation in the Intestinal Tract

Manuscript received 11 March 1997. Initial reviews completed 25 April 1997. Revision accepted 26 September 1997.

Meyer J. Wolin^*, , Yongchao Zhang, Shelton Bank, Susan Yerry^*, and Terry L. Miller^*

^* Wadsworth Center for Laboratories and Research, New York State Department of Health, Empire State Plaza, Albany, NY 12201-0509 and  Department of Chemistry, State University of New York-Albany, Albany, NY 12222

The gastrointestinal tracts of breast-fed infants are colonized more easily with bifidobacteria than are those of formula-fed infants. Colonization is thought to reduce infant diarrhea. Amendments to formulas that improve colonization by bifidobacteria are being actively investigated. Colonization studies almost invariably require measurements of the concentration of the bifidobacteria in feces to assess their importance in the colon. We investigated the use of nuclear magnetic resonance (NMR) analysis of products of fermentations of 1- and 3-¹³C-glucose to evaluate the importance of bifidobacteria in the colonic ecosystem. Bifidobacteria use a unique pathway of hexose catabolism to produce primarily acetate and lactate. The fermentation yields 3 mol of acetate from 2 mol of glucose. Two of the acetates are formed from C1 and C2 of glucose and the third is formed entirely from C3 of glucose. We first employed high resolution NMR to verify the pathway used by a pure culture of Bifidobacterium bifidum. The major products of fermentation of 1- and 3-¹³C-glucose were acetate and lactate. Most of the ¹³C from 3-¹³C-glucose was in ¹³CH₃¹³COOH with equal enrichment in the methyl and carboxyl groups. The ¹³C-acetate from 1-¹³C-glucose was almost entirely enriched in the methyl of acetate and no ¹³CH₃¹³COOH was produced. NMR analysis of glucose fermentation by the colonic flora of a 158-d-old strictly breast-fed infant showed production of ¹³CH₃¹³COOH from 3-¹³C-glucose. The amount of ¹³CH₃¹³COOH formed established that the Bifidobacterium pathway was the major pathway used for glucose fermentation by this infant's colonic microbes.

The Journal of Nutrition Vol. 128 No. 1 January 1998, pp. 91-96
Copyright ©1998 by the American Society for Nutritional Sciences

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