Assaying for thermostable DNA polymerase activity in putative theromophilic bacteria isolates from selected garbage dumping sites and hot springs, in Uganda

Abstract

Molecular and biotechnological research and training in Uganda is hampered by the high cost of acquisition of the required/appropriate chemicals and reagents, notably the DNA polymerase enzyme. Subsequently, there is insufficient practical training in molecular techniques, especially the Polymerase Chain Reaction (PCR) based, at tertiary institutions. This study set out to explore bacterial thermophiles from selected compost pits and hot springs in Uganda for production of thermostable polymerases as an alternative source for use in practical training and biotechnology. The study assessed the basic bacteria cell morphology and colony characteristics; and utilized genotypic methods of 16sRNA sequencing together with Naїve Bayesian Classification to offer a statistical test to flag taxa differentially represented between samples. Putative novel bacteria were isolated from selected garbage compost sites and geothermal hot springs in Uganda. The genotyping revealed that an isolate from compost UG-Wampewo belonged to Proteobacterium at 60% degree of confidence; while the hot spring isolates, Ug Kiga_Mud2,and Ug Kiga, are probably a novel species under Genus Aneurinibacillus (80 % degree of confidence) and Genus Pantoea (47%) respectively. Isolate Ug Kiga_Mud1 is most likely to be a strain of Aneurinibacillus thermoaurophillus as indicated by a degree of confidence at 100%. Characterization for polymerase enzyme activity revealed that the crude protein extracted from compost isolates showed negative results when used in PCR while the crude protein extract from the geothermal hot springs showed positive results. The phenotypic characterization of Ug-Wampewo indicated that this putative novel bacterium is a gram negative coccobacillus with very large cream colonies on nutrient agar. The study recommends comprehensive physiological, biochemical and chemotaxonomic characterization of the novel putative organisms. In addition, a detailed assessment of the organisms is recommended in order to explore the potential applications of the novel putative bacteria in industry and biotechnology.