Assay development and purification of 2-(2'-hydroxyphenyl) benzenesulfinate (HPBS) desulfinase
All fossil fuels are composed primarily of carbon and hydrogen, but also contain sulfur and nitrogen. The presence of sulfur contributes to the corrosion of pipelines, production and refining equipment. When fossil fuels are combusted, they release toxic sulfur oxides that lead to the formation of acid rain; therefore desulfurization of fossil fuels has become of great interest. Bacterial degradation is a newly developed method of desulfurization. A bacterium that is effective in desulfurization of fossil fuels is Rhodococcus sp. IGTS8. Rhodococcus sp. IGTS8 removes sulfur from the model organosulfur compound dibenzothiophene (DBT) without destroying the hydrocarbon backbone using a four enzyme pathway. The focus of this research is the final enzyme in this four enzyme pathway, 2-(2'-hydroxyphenyl) benzenesulfinate desulfinase (HPBS desulfinase). HPBS desulfinase is the slowest enzyme in the desulfurization of DBT. Since it is the slowest of the four enzymes, it is the probable rate-limiting enzyme in vivo. In this study, assays were developed and the partial purification of HPBS desulfinase from Rhodococcus sp. IGTS8 was examined. Use of an anion exchange column and a hydrophobic interaction column allowed partial purification of HPBS desulfinase to be accomplished. Gibb's assay, ultraviolet and visible spectroscopy, fluorescence spectroscopy, and polyacrylamide gel electrophoresis were employed to monitor the purification process.
toxins, fossil fuels, purification, sulfur
Schneider, D. M. (2000). Assay development and purification of 2-(2'-hydroxyphenyl) benzenesulfinate (HPBS) desulfinase (Unpublished thesis). Southwest Texas State University, San Marcos, Texas.