Identification of M. tuberculosis Enzymes Expressed Under Dormant Growth Conditions

Brent Waibel Butler University
Faculty Sponsor(s): Jeremy Johnson Butler University
Mycobacterium tuberculosis is responsible for the most deaths of any one single infectious agent in the world. This research investigated the transition of M. tuberculosis between its active and dormant state and the enzymes required to complete this transition. The main target of this research was to identify enzymes (lipases and esterases) used by M. tuberculosis for energy production in the dormant state. This was accomplished by first inducing dormant conditions to M. smegmatis, a nontoxic relative of M. tuberculosis. Next, the total enzymes from M. smegmatis were isolated and separated based on their size and charge using Native-PAGE gel electrophoresis. Once the enzymes were separated, the esterase and lipase enzymes were isolated by using a fluorescent sensor that only activated in the presence of these two classes of enzymes. The relative amounts of these enzymes between normal and dormant growth conditions were then examined and compared by their relative sensor intensity on the gel. Finally, the identification of the enzymes that were activated under dormant conditions was accomplished by cutting the segments out of the gel and analyzing them by mass spectrometry. This research has long-term implications in the identification of novel drug targets for dormant M. tuberculosis infections.
Biochemistry & Molecular Biology
Oral Presentation

When & Where

10:30 AM
Gallahue Hall 105