Journal: Glycobiology

Article Title: Efficient mucin O- glycan degradation by specific mucin degrading intestinal bacteria: towards understanding enzyme-glycan interactions

doi: 10.1093/glycob/cwag004

Figure Lengend Snippet: Heatmap representing glycoside hydrolyse (GH) and sulphatase as identified by proteomics produced by monocultures A. muciniphila (Am), R. torques (Rt), B. thetaiotaomicron (Bt), co-cultures thereof (AmRt, AmBt, RtBt, and AmRtBt), and the MDSC. Results are based on proteomic analysis (triplicate) of bacterial lysates resulting from cultures grown on PGM for 24 h. the values in the heatmap are the mean of log2 transformed label free quantification (LFQ) intensities of the sum of enzymes that belong to a certain enzyme family. GHs are grouped by associated enzyme activity based on literature ( Montanier et al. 2009 ; Ficko-Blean et al. 2012 ; Etzold et al. 2014 ; Nakamura et al. 2017 ; Drula et al. 2022 ; Wardman et al. 2022 ; Labourel et al. 2023 ). The scale ranges from 0–9 indicating enzyme expression. 0 (orange) indicates enzyme not expressed and 9 (dark blue) indicates enzyme expressed.

Article Snippet: The MDSC contained Akkermansia muciniphila DSM 22959, Ruminococcus gnavus ATCC 29149, Ruminococcus torques ATCC 27756, Bacteroides fragilis DSM 2151, Bacteroides thetaiotaomicron DSM 2079, Phocaeicola vulgatus ATCC 8482, Bacteroides caccae DSM 19024, Anaerostipes caccae DSM 14662, Agathobacter rectalis ATCC 33656, Faecalibacterium duncaniae DSM 17677, Roseburia intestinalis DSM 14610, Desulfovibrio piger DSM 749, and Blautia hydrogenotrophica DSM 10507.

Techniques: Produced, Transformation Assay, Quantitative Proteomics, Activity Assay, Expressing

Journal: Glycobiology

Article Title: Efficient mucin O- glycan degradation by specific mucin degrading intestinal bacteria: towards understanding enzyme-glycan interactions

doi: 10.1093/glycob/cwag004

Figure Lengend Snippet: Overview of the degradation of tested neutral core HMOs, fucosylated (neutral) HMOs, and sialylated (acidic) HMOs by enzymes of bacterial lysates from A. muciniphila (Am), R. torques (rR), B. thetaiotaomicron (Bt), and the mucin-degrading synthetic community (MDSC) after growth on porcine gastric mucin (PGM) for 12 h. degradation is expressed by cleavage of linkages present in the HMOs measured by formation and degradation of intermediate products. Linkage cleavage within 24 h is indicated with “+,” linkage not cleaved within 24 h is indicated with “–,” and no formation of the intermediate is indicated with “not detected (ND)”. This overview is based on results obtained from HPAEC analysis, the chromatograms of which are shown in .

Article Snippet: The MDSC contained Akkermansia muciniphila DSM 22959, Ruminococcus gnavus ATCC 29149, Ruminococcus torques ATCC 27756, Bacteroides fragilis DSM 2151, Bacteroides thetaiotaomicron DSM 2079, Phocaeicola vulgatus ATCC 8482, Bacteroides caccae DSM 19024, Anaerostipes caccae DSM 14662, Agathobacter rectalis ATCC 33656, Faecalibacterium duncaniae DSM 17677, Roseburia intestinalis DSM 14610, Desulfovibrio piger DSM 749, and Blautia hydrogenotrophica DSM 10507.

Techniques:

Journal: Glycobiology

Article Title: Efficient mucin O- glycan degradation by specific mucin degrading intestinal bacteria: towards understanding enzyme-glycan interactions

doi: 10.1093/glycob/cwag004

Figure Lengend Snippet: Overview of the degradation of tested sulphated glycans by enzymes of bacterial lysates from A. muciniphila (Am), R. torques (Rt), B. thetaiotaomicron (Bt), and the mucin-degrading synthetic community (MDSC) after growth on porcine gastric mucin (PGM) for 12 h. degradation is expressed by cleavage of linkages present in the sulphated glycans as measured by degradation of the sulphated glycans and formation of intermediate products. Linkage cleavage within 24 h is indicated with “+” and linkage not cleaved within 24 h is indicated with “–.” this overview is based on the obtained results from PGC-LC-MS/MS analysis of which the chromatograms are shown in – .

Article Snippet: The MDSC contained Akkermansia muciniphila DSM 22959, Ruminococcus gnavus ATCC 29149, Ruminococcus torques ATCC 27756, Bacteroides fragilis DSM 2151, Bacteroides thetaiotaomicron DSM 2079, Phocaeicola vulgatus ATCC 8482, Bacteroides caccae DSM 19024, Anaerostipes caccae DSM 14662, Agathobacter rectalis ATCC 33656, Faecalibacterium duncaniae DSM 17677, Roseburia intestinalis DSM 14610, Desulfovibrio piger DSM 749, and Blautia hydrogenotrophica DSM 10507.

Techniques: Liquid Chromatography with Mass Spectroscopy

Journal: Glycobiology

Article Title: Efficient mucin O- glycan degradation by specific mucin degrading intestinal bacteria: towards understanding enzyme-glycan interactions

doi: 10.1093/glycob/cwag004

Figure Lengend Snippet: (A) Heatmap showing the degradation of O- glycans released from porcine gastric mucin (PGM) after 0, 4, 8, and 24 h incubation with bacterial lysates of A. muciniphila (Am), R. torques (Rt), B. thetaiotaomicron (Bt), co-cultures thereof (AmRt, AmBt, RtBt, and AmRtBt), and the MDSC after growth on PGM for 12 h (average of three replicates). The value at 0 h is set to 100% (log2 scale value 2) and the values at the following timepoints are shown relative to this value (log2 scale; 0 (orange) degraded and 2 or higher (blue) not degraded). (B) Bar graphs showing the relative amount of neutral, sulphated, sialylated, and fucosylated O- glycans released from PGM after 0, 4, 8, and 24 h incubation with bacterial lysates of Am, rt, Bt, AmRt, AmBt, RtBt, AmRtBt, and the MDSC after growth on PGM for 12 h. (C) Bar graphs showing the relative amount of core 1, core 2, core 3, and core 4 O- glycans released from PGM after 0, 4, 8, and 24 h incubation with bacterial lysates of Am, rt, Bt, AmRt, AmBt, RtBt, AmRtBt, and the MDSC after growth on PGM for 12 h.

Article Snippet: The MDSC contained Akkermansia muciniphila DSM 22959, Ruminococcus gnavus ATCC 29149, Ruminococcus torques ATCC 27756, Bacteroides fragilis DSM 2151, Bacteroides thetaiotaomicron DSM 2079, Phocaeicola vulgatus ATCC 8482, Bacteroides caccae DSM 19024, Anaerostipes caccae DSM 14662, Agathobacter rectalis ATCC 33656, Faecalibacterium duncaniae DSM 17677, Roseburia intestinalis DSM 14610, Desulfovibrio piger DSM 749, and Blautia hydrogenotrophica DSM 10507.

Techniques: Incubation

Journal: Glycobiology

Article Title: Efficient mucin O- glycan degradation by specific mucin degrading intestinal bacteria: towards understanding enzyme-glycan interactions

doi: 10.1093/glycob/cwag004

Figure Lengend Snippet: (A) Heatmap showing the degradation of O- glycans released from bovine submaxillary glands (BSM) after 0, 4, 8, and 24 h incubation with bacterial lysates of A. muciniphila (Am), R. torques (Rt), B. thetaiotaomicron (Bt), co-cultures thereof (AmRt, AmBt, RtBt, and AmRtBt), and the MDSC after growth on PGM for 12 h (average of three replicates). The value at 0 h is set to 100% (log2 scale value 2) and the values at the following timepoints are shown relative to this value (log2 scale; 0 (orange) degraded and 2 or higher (blue) not degraded). (B) Bar graph of the ratio between present neutral, sulphated, sialylated, and fucosylated O- glycans released from BSM after 0, 4, 8, and 24 h incubation with bacterial lysates of Am, rt, Bt, AmRt, AmBt, RtBt, AmRtBt, and the MDSC after growth on PGM for 12 h. (C) Bar graphs of the ratio between present core 1, core 2, core 3, and core 4 O- glycans released from BSM after 0, 4, 8, and 24 h incubation with bacterial lysates of Am, rt, Bt, AmRt, AmBt, RtBt, AmRtBt, and the MDSC after growth on PGM for 12 h.

Article Snippet: The MDSC contained Akkermansia muciniphila DSM 22959, Ruminococcus gnavus ATCC 29149, Ruminococcus torques ATCC 27756, Bacteroides fragilis DSM 2151, Bacteroides thetaiotaomicron DSM 2079, Phocaeicola vulgatus ATCC 8482, Bacteroides caccae DSM 19024, Anaerostipes caccae DSM 14662, Agathobacter rectalis ATCC 33656, Faecalibacterium duncaniae DSM 17677, Roseburia intestinalis DSM 14610, Desulfovibrio piger DSM 749, and Blautia hydrogenotrophica DSM 10507.

Techniques: Incubation

Journal: Glycobiology

Article Title: Efficient mucin O- glycan degradation by specific mucin degrading intestinal bacteria: towards understanding enzyme-glycan interactions

doi: 10.1093/glycob/cwag004

Figure Lengend Snippet: Overview of results obtained from proteomics and enzymatic experiments regarding the cleavage of glycan linkages present in HMOs, sulphated glycans, and released O- glycans from PGM and BSM by enzymes of bacterial lysates of A. muciniphila , R. torques , and B. thetaiotaomicron , in monocultures, co-cultures, and the MDSC. Linkage cleavage within 24 h is indicated with “+,” linkage not cleaved within 24 h is indicated with “–,” and not applicable/not tested is indicated with “NA.”

Article Snippet: The MDSC contained Akkermansia muciniphila DSM 22959, Ruminococcus gnavus ATCC 29149, Ruminococcus torques ATCC 27756, Bacteroides fragilis DSM 2151, Bacteroides thetaiotaomicron DSM 2079, Phocaeicola vulgatus ATCC 8482, Bacteroides caccae DSM 19024, Anaerostipes caccae DSM 14662, Agathobacter rectalis ATCC 33656, Faecalibacterium duncaniae DSM 17677, Roseburia intestinalis DSM 14610, Desulfovibrio piger DSM 749, and Blautia hydrogenotrophica DSM 10507.

Techniques: Glycoproteomics