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 Citations (Scopus)

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

Fingerprint

Oligosaccharides
Bacteria
Glycoside Hydrolases
Galactosidases
Enzymes
Human Milk

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

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

Identification and accurate quantitation of biological oligosaccharide mixtures. / Strum, John S.; Kim, Jaehan; Wu, Shuai; De Leoz, Maria Lorna A; Peacock, Kyle; Grimm, Rudolf; German, J. Bruce; Mills, David A.; Lebrilla, Carlito B.

In: Analytical Chemistry, Vol. 84, No. 18, 18.09.2012, p. 7793-7801.

Research output: Contribution to journalArticle

Strum, JS, Kim, J, Wu, S, De Leoz, MLA, Peacock, K, Grimm, R, German, JB, Mills, DA & Lebrilla, CB 2012, 'Identification and accurate quantitation of biological oligosaccharide mixtures', Analytical Chemistry, vol. 84, no. 18, pp. 7793-7801. https://doi.org/10.1021/ac301128s
Strum JS, Kim J, Wu S, De Leoz MLA, Peacock K, Grimm R et al. Identification and accurate quantitation of biological oligosaccharide mixtures. Analytical Chemistry. 2012 Sep 18;84(18):7793-7801. https://doi.org/10.1021/ac301128s
Strum, John S. ; Kim, Jaehan ; Wu, Shuai ; De Leoz, Maria Lorna A ; Peacock, Kyle ; Grimm, Rudolf ; German, J. Bruce ; Mills, David A. ; Lebrilla, Carlito B. / Identification and accurate quantitation of biological oligosaccharide mixtures. In: Analytical Chemistry. 2012 ; Vol. 84, No. 18. pp. 7793-7801.
@article{c94a2d84f3af46828010ad307844c2cb,
title = "Identification and accurate quantitation of biological oligosaccharide mixtures",
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.",
author = "Strum, {John S.} and Jaehan Kim and Shuai Wu and {De Leoz}, {Maria Lorna A} and Kyle Peacock and Rudolf Grimm and German, {J. Bruce} and Mills, {David A.} and Lebrilla, {Carlito B}",
year = "2012",
month = "9",
day = "18",
doi = "10.1021/ac301128s",
language = "English (US)",
volume = "84",
pages = "7793--7801",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "American Chemical Society",
number = "18",

}

TY - JOUR

T1 - Identification and accurate quantitation of biological oligosaccharide mixtures

AU - Strum, John S.

AU - Kim, Jaehan

AU - Wu, Shuai

AU - De Leoz, Maria Lorna A

AU - Peacock, Kyle

AU - Grimm, Rudolf

AU - German, J. Bruce

AU - Mills, David A.

AU - Lebrilla, Carlito B

PY - 2012/9/18

Y1 - 2012/9/18

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84866345911&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84866345911&partnerID=8YFLogxK

U2 - 10.1021/ac301128s

DO - 10.1021/ac301128s

M3 - Article

C2 - 22897719

AN - SCOPUS:84866345911

VL - 84

SP - 7793

EP - 7801

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 18

ER -