Identification and accurate quantitation of biological oligosaccharide mixtures

John S. Strum, Jaehan Kim, Shuai Wu, Maria Lorna A De Leoz, Kyle Peacock, Rudolf Grimm, J. Bruce German, David A. Mills, Carlito B Lebrilla

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

Structure-specific characterization and quantitation is often required for effective functional studies of oligosaccharides. Inside the gut, HMOs are preferentially bound and catabolized by the beneficial bacteria. HMO utility by these bacteria employs structure-specific catabolism based on a number of glycosidases. Determining the activity of these enzymes requires accurate quantitation of a large number of structures. In this study, we describe a method for the quantitation of human milk oligosaccharide (HMO) structures employing LC/MS and isotopically labeled internal standards. Data analysis was accomplished with a newly developed software tool, LC/MS Searcher, that employs a reference structure library to process LC/MS data yielding structural identification with accurate quantitation. The method was used to obtain a meta-enzyme analysis of bacteria, the simultaneous characterization of all glycosidases employed by bacteria for the catabolism of milk oligosaccharides. Analysis of consumed HMO structures confirmed the utility of a β-1,3-galactosidase in Bifidobacterium longum subsp. infantis ATCC 15697 (B. infantis). In comparison, Bifidobacterium breve ATCC 15700 showed significantly less HMO catabolic activity compared to B. infantis.

Original languageEnglish (US)
Pages (from-to)7793-7801
Number of pages9
JournalAnalytical Chemistry
Volume84
Issue number18
DOIs
StatePublished - Sep 18 2012

ASJC Scopus subject areas

  • Analytical Chemistry

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    Strum, J. S., Kim, J., Wu, S., De Leoz, M. L. A., Peacock, K., Grimm, R., German, J. B., Mills, D. A., & Lebrilla, C. B. (2012). Identification and accurate quantitation of biological oligosaccharide mixtures. Analytical Chemistry, 84(18), 7793-7801. https://doi.org/10.1021/ac301128s