Structural and Enzymatic Comparison of Faecalibacterium prausnitzii GH31 α-Glycosidases
dc.contributor.author | Jewczynko, Anna | |
dc.date.accessioned | 2022-05-02T12:58:41Z | |
dc.date.available | 2022-05-02T12:58:41Z | |
dc.date.issued | 2022-05-02 | |
dc.date.submitted | 2022-04-06 | |
dc.description.abstract | The gut microbiome is home to thousands of species of bacteria, that are essential for human digestion, immunity and physiology. Faecalibacterium prausnitzii makes up about 5% of a healthy human gut microbiome and a lower abundance of this bacterium has been found in patients with IBD and Crohn’s disease. Among an extensive repertoire of carbohydrate active enzymes, F. prausnitzii has 2 GH31 enzymes, which are from the same family as Sucrase-Isomaltase and Maltase-Glucoamylase, human digestive enzymes with overlapping and distinguishing substrate specificities. This thesis aims to characterize the substrate specificity, preference and inhibition sensitivity of F. prausnitzii GH31 α-glucosidases to better understand the structural features of GH31 enzymes and the biological capabilities of these bacteria. In this thesis, AlphaFoldV2.1.0 was used to create computational models of F. prausnitzii α-glucosidases, and the substrate specificity, enzyme kinetics and inhibition parameters are reported. Structurally, these α-glucosidases have the same identified conserved N-terminal and (β/α)8 barrel domains, but FpAG1 has an additional conserved domain of unknown function at the C-terminus which is not found in the FpAG2 structure. Both FpAG1 and FpAG2 have α-glucosidase and oligo-1,6-glucosidase activity. The comparative kinetic studies show that FpAG1 has a greater preference for α-1,6 glycosidic linkages, and FpAG2 has a greater preference for α-1,4 glycosidic linkages. The comparative inhibition studies show that the tested α-glucosidase inhibitors, acarbose, miglitol and kotalanol, are more potent in FpAG2 than FpAG1, and acarbose is a weak inhibitor of F. prausnitzii α-glucosidases. Distinguishing binding affinities of miglitol and kotalanol in these GH31 enzymes suggest structural differences in the FpAG1 and FpaG2 active sites. Gaining insight on the GH31 α-glucosidases as a component of F. prausnitzii metabolism can further our understanding of this community in the human gut microbiome. | en |
dc.identifier.uri | http://hdl.handle.net/10012/18212 | |
dc.language.iso | en | en |
dc.pending | false | |
dc.publisher | University of Waterloo | en |
dc.title | Structural and Enzymatic Comparison of Faecalibacterium prausnitzii GH31 α-Glycosidases | en |
dc.type | Master Thesis | en |
uws-etd.degree | Master of Science | en |
uws-etd.degree.department | Biology | en |
uws-etd.degree.discipline | Biology | en |
uws-etd.degree.grantor | University of Waterloo | en |
uws-etd.embargo.terms | 0 | en |
uws.contributor.advisor | Rose, David | |
uws.contributor.affiliation1 | Faculty of Science | en |
uws.peerReviewStatus | Unreviewed | en |
uws.published.city | Waterloo | en |
uws.published.country | Canada | en |
uws.published.province | Ontario | en |
uws.scholarLevel | Graduate | en |
uws.typeOfResource | Text | en |