Team:Duke/Results

Does BadA Work?

In order to characterize our enzyme, it was necessary to determine if the enzyme had actually worked. This was done by graphing the data from the BadA lysate and the Empty Vector lysate (see graph below). In the graph below, it is possible to see that the absorption within the BadA lysate went down farther and faster than the Empty Vector lysate. This indicates that the enzyme is active and is working.

Another part of determining if the test was valid was to compare the controls to the sample of interest. This was done by graphing the samples from the above graph with their respective controls (see the following two graphs). These graphs indicate that the CoA was being registered as the trial without CoA added did not have any absorbance. We can also determine that the cells were active because the samples without ATP were functioning but not at the rate that the full ingredient sample does. Finally, because the trial where full ingredients were used above the trial where benzoic acid was removed means that the benzoic acid was being utilized by the enzyme.

Varying the Amount of Benzoic Acid

After confirming that the BadA enzyme is working, it is necessary to investigate the absorption of the Ellman’s Reagent when the amount of benzoic acid was varied. In the experiment the benzoic acid was varied through the following values: 0.05, 0.25, 0.5, 0.75, and 1 mM. The graph of these trials can be seen in the graph below.