Investigating the Possible Role of a Glycosyl Transferase Protein in the Biosynthesis of Long-Chain Hydrocarbons in Shewanella oneidensis

Abstract

In the search for alternative sources of energy, new organisms are being looked at as potential biofuel producers. It has been shown that the longchain hydrocarbons produced by the bacterium Shewanella oneidensis can be broken down into usable fuel. It is known that the enzyme OleA is largely responsible for hydrocarbon synthesis, but other proteins may also play a role. In my thesis work, I deleted the gene SO_3174, which encodes a glycosyl transferase protein and was found to be interrupted in a transposon screen for increased hydrocarbon production, from S. oneidensis MR-1. I then tried to show that deleting SO_3174 resulted in an increase in hydrocarbon production just as the interruption of the gene had. The deletion strain showed an increased fluorescence in the presence of Nile Red dye, a hydrophobic dye that can be used to indirectly detect hydrocarbon levels. However, the deletion strain did not exhibit increased hydrocarbons during direct analysis of nonpolar extractions. These same results were obtained from a strain containing both the SO_3174 deletion and Stenotrophomonas maltophilia OleA. Overall, I found that the protein encoded by SO_3174 most likely does not play a role in hydrocarbon biosynthesis in S. oneidensis.