The gut microbiota, comprised of a plethora of bacterial species, is a crucial part of the digestive system. The intestinal flora regulates many physiological processes and contributes to a large proportion of an individual's daily energy production. Certain bacteria phyla that reside in the mucus layer of the large intestine play an important role in our physiology. Gut bacteria are capable of processing complex fibres that are present in our diet, influencing organ function, and modulating the immune system. Even slight changes in the microbiota’s composition result in several pathologies. Bacteroides species are known for their ability to metabolize carbohydrates and have acquired interesting mechanisms of processing O- and N-glycosylated proteins, with the latter being the focus of this study. This experiment aims to elucidate the degrading mechanisms of N-glycosylated proteins by Bacteroides species, differentiate between the main degrader species, and analyse the extent of this degradation. Out of all the bacteria strains tested, Bacteroides fragilis and Bacteroides massiliensis displayed proteolytic activity on fetuin, which was further verified through Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) and thin-layer chromatography (TLC) analyses. Additionally, six B.fragilis proteins from the GH18 family were cloned and tested for enzymatic activity on various substrates, however, no activity was observed. This work established the phenotypes of bacterial N-glycan degradation.
| Identifer | oai:union.ndltd.org:UPSALLA1/oai:DiVA.org:his-23988 |
| Date | January 2024 |
| Creators | Lani, Magdalena |
| Publisher | Högskolan i Skövde, Institutionen för hälsovetenskaper |
| Source Sets | DiVA Archive at Upsalla University |
| Language | English |
| Detected Language | English |
| Type | Student thesis, info:eu-repo/semantics/bachelorThesis, text |
| Format | application/pdf |
| Rights | info:eu-repo/semantics/openAccess |
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