Fig. 5: Surface activity of mucin-grown A. muciniphila against TriLacNAc.
aA. muciniphila cells were grown on PGM III, harvested and TriLacNAc added to assess surface enzyme activity. Samples were analysed at different time points following addition of TriLacNAc. Equivalent data for the Bacteroides species are shown in Supplementary Fig. 13. b Samples from two different time points of the A. muciniphila whole cell assay were incubated with two different exo-acting enzymes of known specificity to identify the products. The asterisks indicate the time points analysed. The top and bottom of the glycan structures shown is the non-reducing and reducing end, respectively. A: original sample, B: +β1,4-galactosidase BT0461GH2, C: +broad-acting β-GlcNAc’ase BT0459GH20. For example, in both time points, the two different types of trisaccharides can be seen to be hydrolysed in the separate digests: (1) Galβ1,4GlcNAcβ1,3Gal disappears in ‘B’ and products Gal and GlcNAcβ1,3Gal are now present, (2) GlcNAcβ1,3Galβ1,4GlcNAc disappears in ‘C’ and products GlcNAc and Galβ1,4GlcNAc are now present. These experiments were carried out once, but multiple pilot experiments were run and were consistent with the data shown. The source data are provided in the source data file.
