Characterization of Mycobacterium tuberculosis isoprenyl diphosphate synthase Rv2173 Faith Adekunle, Dr. Francis M. Mann
Mycobacterium tuberculosisis a species of pathogenic bacteria in the family of Mycobacteriaceae and the causative agent for the human disease Tuberculosis. This pathogen is responsible for approximately 1 million deaths every year. The bacteria is known to develop resistance to common antibacterial agents. This resistance mechanism limits the application of available antibiotics to treat it . A potential target for this is the biosynthetic machinery for terpenoids; these molecules are involved in energy production, protection of the bacteria, cell wall biosynthesis, and modulation of the human immune system.
While much of the terpenoid biosynthetic enzymes have been characterized, Rv2173 has unknown function. Structural data has been acquired, and the enzyme appears to be homologous to previously characterized trans-isoprenyl diphosphate synthases. Yet, the function remains elusive. This may be due to a non-canonical shallow and wide active site that may provide for a novel drug target, however, characterization of the enzyme is required to confirm this hypothesis.
The bacteria was cloned into an expression vector containing an N-terminal 6xHis tag and expressed in Escherichia coli C41 cells and then puried using affinity chromatography and confirmed via SDS-PAGE, then enzymatic assays were made to check enzyme activity with varying allylic substrates. Gas chromatography showed presence of the 15-carbon product, FPP.
