Abstract

IGT has developed a microbial culture of Rhodococcus rhodochrous, designated as IGTS8, that is capable of specifically cleaving carbon-sulfur bonds in a range of organosulfur model compounds and is capable of removing organic sulfur from coal and petroleum without significantly sacrificing the calorific value of the fuel. Although IGTS8 possesses the ability to specifically remove organic sulfur from coal, a major research need is to develop improved strains of microorganisms that possess higher levels of desulfurization activity and therefore will permit more favorable biodesulfurization process conditions: faster rates, more complete removal, and smaller reactor size. Strain improvement is the single most important aspect to the development of a practical coal biodesulfurization process and accordingly is the focus of research in this project. During the past year, significant progress was made toward improving the biodesulfurization capabilities of Rhodococcus Rhodochrous IGTS8. The main objective was to identify and characterize strong promoters of IGTS8. The DNA sequencing of the promoter region and chloramphenicol resistance gene of pRF2, as well as six mutant promoters, was determined. The 16S structural gene of IGTS8 was isolated and used to identify the putative promoter of this gene. Four promoter probe vectors were constructed and are currently being used to analyze the strength of Rhodococcus promoters: from the IGTS8 genome, mutants of promoters from the chloramphenicol resistance gene of pRF2, the promoter from the 16S RNA gene, and various strong inducible promoters.

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