Phytogenetic analysis by 16s rRNA gene sequence of a protease-secreting thermophilic bacterial isolate KITNT-3 from Kiteezi landfill

dc.contributor.author Nabachwa, Rita
dc.date.accessioned 2014-10-28T11:34:19Z
dc.date.available 2014-10-28T11:34:19Z
dc.date.issued 2011-07
dc.description A Thesis submitted to the School of Graduate Studies in partial fulfillment of the requirements for the award of the Degree of Master of Science in Chemistry of Makerere University en_US
dc.description.abstract A thermophilic protease-secreting bacterial isolate KITNT-3 from Kiteezi landfill was characterized phylogenetically by determining its relationship with other existing known bacterial strains. Genomic DNA was isolated from the cells and the 16S rRNA gene amplified using universal primers. The amplified PCR product was purified using an elaborate ethanol/EDTA precipitation method and subjected to sequence analysis using an ABI 3130XL genetic analyzer. In order to determine the phylogenetic position of the isolate, the partial 16S rRNA gene sequence was compared with other known 16S rRNA gene sequences of related taxa from the GenBank. A phylogenetic tree was constructed using the MEGA v4 program. The analysis showed that the isolate clusters with members of the genus Bacillus and was closest to Bacillus thuringiensis strain T13001 (ACNC01000278), sharing 99.8 % nucleotide sequence similarity. The crude enzyme was obtained from the culture filtrate of the isolate after an overnight incubation at 550C in a shaking incubator at 80 rpm and the extract was characterized for its caseinolytic activity. The morphological and growth characteristics showed that isolate KITNT-3 is an aerobic, Gram-positive, endospore-forming, thermophile with rod-shaped cells measuring 3.2 µm in length and 0.8 µm in width respectively. The crude enzyme was found to exhibit a specific caseinolytic activity of 0.0065 µmoles tyrosine equivalents/min/mg protein at optimal assay conditions of pH 6 and temperature of 550C. Further purification and characterization of the crude enzyme solution is deemed necessary for a comprehensive enzyme classification. en_US
dc.identifier.citation Nabachwa, R. (2011). Phytogenetic analysis by 16s rRNA gene sequence of a protease-secreting thermophilic bacterial isolate KITNT-3 from Kiteezi landfill (Unpublished master's thesis). Makerere University, Kampala, Uganda en_US
dc.identifier.uri http://hdl.handle.net/10570/4049
dc.language.iso en en_US
dc.publisher Makerere University en_US
dc.subject Bacterial isolate KITNT-3 en_US
dc.subject Protease-secreting thermophyle en_US
dc.subject 16s rRNA gene en_US
dc.subject Sequence en_US
dc.subject Phytogenetic analysis en_US
dc.subject Kiteezi landfill, Kampala, Uganda en_US
dc.title Phytogenetic analysis by 16s rRNA gene sequence of a protease-secreting thermophilic bacterial isolate KITNT-3 from Kiteezi landfill en_US
dc.type Thesis en_US
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