Master Degree / Yüksek Lisans Tezleri
Permanent URI for this collectionhttps://hdl.handle.net/11147/3008
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Master Thesis Xylan Degradation Mechanism of Human Intestinal Bacteria(Izmir Institute of Technology, 2017) Polat, Nüket; Büyükkileci, Ali Oğuz; Güleç, ŞükrüXylan is the second most abundant plant cell wall polysaccharide after cellulose. The xylan rich lignocellulosic material obtained from agriculture, forestry and industrial wastes provides cost effective raw materials. The degradation of xylan in the human body is an important process contributing to the continuation of the microbial communities living in the human colonic ecosystem. Due to its complex, long chain structure and the various chemical bonds it contains, xylan hydrolysis requires different enzymatic activities. Bacteria that live in the colon and are useful for human health, such as Bifidobacterium and Lactobacillus species can not perform xylan utilization. However, several types of xylan are utilized by the Bacteroides species, which have the second largest density in the colon. In this study, different Bifidabacterium and Bacteriodes species were investigated for their ability to degrade beechwood xylan and corncob xylan. Bifidabacterium and Bacteriodes were cultured together in tubes containing xylan as the sole carbon source. It was observed that; the B. animalis subsp. lactis, which does not have the ability to use the xylan, could grow when cultured on xylan-containing medium with Bacteroides species. These showed that, the xylan in the media was degraded into xylooligosaccharides by the Bacteroides species and the XOS formed was used as a carbon source by both species. The short chain fatty acid and lactic and succinic acid production profiles of co-cultures were different than the mono cultures, indicating a positive effect of co-culturing. This study showed that xylan is a potential prebiotic carbohydrate, which can selectively stimulate the growth of beneficial bacteria in the colon, as a result of possible cross feeding of different bacteria residing in the colon.Master Thesis Purification and Biochemical Characterization of Xylanase Expressed in Thermophilic Geobacillus Sp.(Izmir Institute of Technology, 2015) Algan, Müge; Köksal, Mustafa; Şanlı Mohamed, Gülşah; Şanlı Mohamed, Gülşah; Köksal, MustafaXylanase is an enzyme that catalyzes the degradation of the linear polysaccharide β-1,4-xylan into xylose and breaks down the hemicellulose structure of plant cell wall. The xylanolytic property of the enzyme makes it preferable for many biotechnological applications in industry. This enzyme is possibly produced by some bacterial and fungal microorganisms. In this study, briefly, xylanase enzyme was expressed in thermophillic Geobacillus sp. and purified by cold acetone precipitation and gel filtration chromatography. Molecular weight of our xylanase was found as 40.1 kDa by SDS-PAGE and this protein band was verified by Native-PAGE activity staining. Finally, it was characterized using biochemical methods. For characterization studies, Km and Vmax values were calculated from Lineweaver-Burk plot as 10.2 mg/ml and 31.7 U/ml, respectively. The optima temperature and pH for enzyme activity were investigated using beechwood xylan as substrate and found as 55°C and 8.0, respectively. Furthermore, effects of some metal ions, various chemical reagents and organic solvents on enzyme activity were also determined and we observed that Ca2+, Mn2+ and Co2+ affected the activity positively while Zn2+, Cd2+, Fe3+, EDTA, SDS, CHAPS and DTT shielded the activity. And only β-mercaptoethanol caused a significant change amoung organic solvents. Lastly, that the enzyme has a long shelf-life was confirmed assaying the samples taken from enzyme stocks stored at +4°C and room temperature for six weeks.