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A versatile and scalable strategy for glycoprofiling bifidobacterial consumption of human milk oligosaccharides.

Locascio RG, Niñonuevo MR, Kronewitter SR, Freeman SL, German JB, Lebrilla CB, Mills DA - Microb Biotechnol (2008)

Bottom Line: For quantitative consumption, deuterated and reduced human milk oligosaccharide (HMO) standards were used.A detailed analysis of consumption glycoprofiles indicated strain-specific capabilities towards differential metabolism of milk oligosaccharides.This method overcomes previous limitations in the quantitative, multi-strain analysis of bacterial metabolism of HMOs and represents a novel approach towards understanding bacterial consumption of complex prebiotic oligosaccharides.

View Article: PubMed Central - PubMed

Affiliation: Departments of Viticulture & Enology, Chemistry and Food Science & Technology, and Microbiology Graduate Group, University of California, Davis, CA 95616, USA.

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HPLC‐Chip/TOF‐MS analysis of selectivity for specific glycan structures consumed by B. infantis ATCC 15697. Shown are nanoLC extracted ion chromatograms (XIC) of selected milk glycans recovered (A) after incubation with B. infantis ATCC 15697 and (B) before incubation (control). The complete disappearance (A) of all glycan peaks for specific oligosaccharide (m/z: 732, 878 and 1243) indicates that B. infantis consumes specific HMOs but does not discriminate for any specific isomers within each group of HMO consumed.
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f4: HPLC‐Chip/TOF‐MS analysis of selectivity for specific glycan structures consumed by B. infantis ATCC 15697. Shown are nanoLC extracted ion chromatograms (XIC) of selected milk glycans recovered (A) after incubation with B. infantis ATCC 15697 and (B) before incubation (control). The complete disappearance (A) of all glycan peaks for specific oligosaccharide (m/z: 732, 878 and 1243) indicates that B. infantis consumes specific HMOs but does not discriminate for any specific isomers within each group of HMO consumed.

Mentions: The highest growth was observed mainly among the B. infantis strains JCM 7009, JCM 7010, JCM 7007, JCM 7011 and ATCC 15697, respectively, for which a nearly complete consumption of all HMOs with a DP (degree of polymerization of the HMOs) ≤ 7 (m/z = 732.25, 878.31, 1097.39 and 1243.44) was observed (Table 1, Figs 2 and S1). Among these strains the HMO consumption patterns resulted in highly similar glycoprofiles and the total HMO utilization ranged between 87.15% and 84.08% (Table 1). Except ATCC 15697, the rest of the strains have been classified by ribotyping and RAPD‐PCR to belong to the infantis biovar (Sakata et al., 2002). Compared with other bifidobacterial species tested these B. infantis strains shows unique glycoprofiling patterns with very high metabolic capacity for HMOs. This is also clearly reflected in their growth kinetic analysis for having distinctly similar growth rates and the highest ODs achieved among all the tested strains. Deuterated and reduced milk oligosaccharides were added as internal standards (Fig. 3) and Glycolyzer was used to perform automatic calculations of D/H ratios which were used as metric for measuring specific oligosaccharide consumption. HPLC‐Chip/TOF‐MS analysis of B. infantis ATCC 15697 oligosaccharide consumption patterns confirmed previous MALDI‐FTICR‐MS data (LoCascio et al., 2007) indicating that among several HMOs available B. infantis consumes only specific glycans. The HPLC‐Chip/TOF‐MS system enabled the detailed HMO consumption analysis at the single isomer level revealing that B. infantis does not discriminate among isomers of the same HMO (Fig. 4).


A versatile and scalable strategy for glycoprofiling bifidobacterial consumption of human milk oligosaccharides.

Locascio RG, Niñonuevo MR, Kronewitter SR, Freeman SL, German JB, Lebrilla CB, Mills DA - Microb Biotechnol (2008)

HPLC‐Chip/TOF‐MS analysis of selectivity for specific glycan structures consumed by B. infantis ATCC 15697. Shown are nanoLC extracted ion chromatograms (XIC) of selected milk glycans recovered (A) after incubation with B. infantis ATCC 15697 and (B) before incubation (control). The complete disappearance (A) of all glycan peaks for specific oligosaccharide (m/z: 732, 878 and 1243) indicates that B. infantis consumes specific HMOs but does not discriminate for any specific isomers within each group of HMO consumed.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3815754&req=5

f4: HPLC‐Chip/TOF‐MS analysis of selectivity for specific glycan structures consumed by B. infantis ATCC 15697. Shown are nanoLC extracted ion chromatograms (XIC) of selected milk glycans recovered (A) after incubation with B. infantis ATCC 15697 and (B) before incubation (control). The complete disappearance (A) of all glycan peaks for specific oligosaccharide (m/z: 732, 878 and 1243) indicates that B. infantis consumes specific HMOs but does not discriminate for any specific isomers within each group of HMO consumed.
Mentions: The highest growth was observed mainly among the B. infantis strains JCM 7009, JCM 7010, JCM 7007, JCM 7011 and ATCC 15697, respectively, for which a nearly complete consumption of all HMOs with a DP (degree of polymerization of the HMOs) ≤ 7 (m/z = 732.25, 878.31, 1097.39 and 1243.44) was observed (Table 1, Figs 2 and S1). Among these strains the HMO consumption patterns resulted in highly similar glycoprofiles and the total HMO utilization ranged between 87.15% and 84.08% (Table 1). Except ATCC 15697, the rest of the strains have been classified by ribotyping and RAPD‐PCR to belong to the infantis biovar (Sakata et al., 2002). Compared with other bifidobacterial species tested these B. infantis strains shows unique glycoprofiling patterns with very high metabolic capacity for HMOs. This is also clearly reflected in their growth kinetic analysis for having distinctly similar growth rates and the highest ODs achieved among all the tested strains. Deuterated and reduced milk oligosaccharides were added as internal standards (Fig. 3) and Glycolyzer was used to perform automatic calculations of D/H ratios which were used as metric for measuring specific oligosaccharide consumption. HPLC‐Chip/TOF‐MS analysis of B. infantis ATCC 15697 oligosaccharide consumption patterns confirmed previous MALDI‐FTICR‐MS data (LoCascio et al., 2007) indicating that among several HMOs available B. infantis consumes only specific glycans. The HPLC‐Chip/TOF‐MS system enabled the detailed HMO consumption analysis at the single isomer level revealing that B. infantis does not discriminate among isomers of the same HMO (Fig. 4).

Bottom Line: For quantitative consumption, deuterated and reduced human milk oligosaccharide (HMO) standards were used.A detailed analysis of consumption glycoprofiles indicated strain-specific capabilities towards differential metabolism of milk oligosaccharides.This method overcomes previous limitations in the quantitative, multi-strain analysis of bacterial metabolism of HMOs and represents a novel approach towards understanding bacterial consumption of complex prebiotic oligosaccharides.

View Article: PubMed Central - PubMed

Affiliation: Departments of Viticulture & Enology, Chemistry and Food Science & Technology, and Microbiology Graduate Group, University of California, Davis, CA 95616, USA.

Show MeSH