Biodesulfurization (BDS) of petroleum has been investigated as an alternative method to conventional chemical desulfurization for many years.... Show moreBiodesulfurization (BDS) of petroleum has been investigated as an alternative method to conventional chemical desulfurization for many years. Despite its potential to be an environmentally benign method, it has not been developed sufficiently to be useful in real world applications. This is due to its low efficiency and the necessity for it to work at temperatures high enough to lower the viscosity of petroleum so that mixing can be achieved. This study places the spotlight on two strains of Paenibacillus isolated in our laboratory that, together, possess biodesulfurization ability at moderately high temperatures and attempts to enhance biodesulfurization by expression of Vitreoscilla hemoglobin (VHb) in the Paenibacillus strains. The effects of expression of the VHb gene (vgb) on growth and desulfurizing activity was examined in a symbiotic system between the Paenibacillus strains 32O-Y and 32O-W. Of the two, 32O-Y is the one with the ability to metabolize dibenzothiophene (DBT), a common compound in petroleum that contains organic sulfur, while 32O-W enhances this ability, forming a symbiotic relationship between the two. The transformant of 32O-Y bearing vgb cloned into the shuttle vector pNW33N had been previously constructed in our laboratory. Presence of pNW33N-vgb was verified in one strain of 32O-Y through isolation of DNA, PCR, and gel electrophoresis. Mixtures of 32O-Y/32O-W or 32O-Y[pNW33N-vgb]/32O-W were cultured in minimal medium (CDM) with DBT as the sole sulfur source and subjected to multiple trials of growth and assay of DBT metabolism at varying temperatures. At 45 ˚C there was a substantial increase in both growth and DBT metabolizing coincident with VHb expression, whereas at lower (37 ˚C) and higher (50 ˚C) temperatures, VHb expression had little to no effect on either parameter. For both growth and DBT metabolism tested at 37 ˚C, 45 ˚C and 50 ˚C the highest absolute levels were seen at 37 ˚C, and the lowest at 45 ˚C. M.S. in Biology, May 2015 Show less
