Expression of the Propanoyl-CoA Metabolic Pathway from T. Fusca in E. Coli

The objective of this project was to successfully express a metabolic pathway found in Thermobifida fusca, a thermophilic, cellulolytic actinobacteria, in the model organism, Escherichia coli.  A potentially novel method for biologically producing 1-propanol was found in an engineered strain of T. fusca.  Direct confirmation of the novel pathway’s activity is difficult in T. fusca, thus expression of the target pathway in E. coli would provide a direct means of testing the novel pathway.  The T. fusca gene, Tfu_2395, was transformed into E. coli and positive transformants were confirmed with blue/white screening and DNA sequencing. Secretion of 1-propanol by the engineered strain of E. coli would functionally demonstrate the activity of novel metabolic pathway for production of 1-propanol found in T. fusca and secondary confirmation will be achieved using molecular measurements such as real-time PCR of pathway genes.  This research was funded by the Virginia Academy of Science and Virginia Commonwealth University.