Master Degree / Yüksek Lisans Tezleri
Permanent URI for this collectionhttps://hdl.handle.net/11147/3008
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Master Thesis Genotypic Characterization of Extracellular Enzyme Producing Thermophilic Bacteria in Balçova Geothermal Region(Izmir Institute of Technology, 2003) Yavuz, Elif; Yenidünya, Ali Fazıl; Yavuz, Elif; Yenidünya, Ali Fazıl; 04.03. Department of Molecular Biology and Genetics; 01. Izmir Institute of Technology; 04. Faculty of ScienceThermophiles are the organisms which are adapted to live at high temperatures. The enzymes from thermophiles find a number of commercial applications because of their thermostability and thermoactivity. Therefore, the isolation of thermophilic bacteria from natural sources and their identification are very important in terms of discovering new industrial enzymes.In keeping with this view, Balçova Geothermal Region could serve as a good source for new thermophilic microorganisms with novel industrially important properties.The aim of this research was therefore the isolation of industrially important extracellular enzyme producing thermophilic bacteria from Balçova Geothermal Region and their identification by genetical means. 16S-ITS rDNA RFLP, plasmid profiling and pulsed field gel electrophoresis studies were performed for this purpose.112 thermophilic strains were isolated from various environmental samples collected within Balçova Geothermal Region. These strains were screened for the existence of 6 extracellular enzyme activities. These were, lipases, amylases, proteases, xylanases, cellulases and pectinases. In total, 110 lipase (tween 20 as substrate), 106 amylase, 55 protease, 28 xylanase, 10 cellulase and 3 pectinase activities were detected.Some other phenotypic tests were also performed for these isolated strains. Since all the isolated strains were Gram (+), endospore forming rods, they were identified as Bacillus sp.16S-ITS rDNA RFLP and plasmid RFLP profiles were produced by using two restriction endonucleases Taq I and Hae III . The isolated strains were clustered into eleven groups by Taq I restriction profiles of 16S-ITS rDNA while nine groups were obtained by Hae III digestion profiles. When these groups were compared, it was concluded that 17 genotypically different strains existed in total 112 isolates. Two of the isolated strains yielded similar RFLP profiles to those of Bacillus stearothermophilus (CECT 43) reference strain.Plasmid profiling was also performed. It was found that 23 of the isolated strains contained plasmid DNA. Hae III restriction profiles indicated the existence of three different types of plasmids.PFGE optimization studies by Sma I restriction endonuclease for thermophilic Bacilli were also performed. A new method for preparation of agarose plugs was developed.Master Thesis Biochemical and Molecular Characterization of Extracellular Enzyme Producing Staphylococci Isolated From Different Origins(Izmir Institute of Technology, 2006) Appak, Sıla; Güneş, Hatice; Güneş, Hatice; 04.03. Department of Molecular Biology and Genetics; 04. Faculty of Science; 01. Izmir Institute of TechnologyStaphylococci are pathogenic bacteria known to cause diseases among diferrent organisms including human. The two species Staphylococcus aureus and Staphylococcus epidermidis are well defined in human diseases although their exact mechanism of pathogenesis is still not fully understood. These pathogenic bacteria could be isolated from soil, water, air, as well as from the living organisms and they are both pathogenic and saprophytic.Extracellular enzymes of the organisms are used for the industrial purposes. The isolation and characterization of these enzymes are crucial steps in biotechnology. The extracellular enzymes derived from the bacteria serve for many purposes in the industry. In this project 128 Staphylococcus sp. were used. Of these 128 bacteria, 12 were isolated from patients, 40 were isolated from the foodhandler.s hygiene detections, 27 were isolated from pygeons and 49 of them were reference strains. They were searched for the presence of some of the industrially important extracellular enzymes: protease, lipase, cellulase, xylanase, amylase, laccase, urease, DNase and pectinase with biochemical tests. They were also searched for the presence of the lipase, protease and thermonuclease amplifications by PCR. The bacteria apart from the refence strains were also tried to be identified by 16S-ITS-rRNA RFLP analysis. The results would indicate the extracellular enzyme production among these pathogenic bacteria and would also be used as a guide in further studies to correlate between Staphylococcal pathogenity and enzyme production.