Team:UFlorida/Results

Results

Liquid cultures of our bacteria were prepared in King’s Medium B supplemented with tryptophan. The culture was centrifuged after growth and the supernatant was filtered and used in HPLC. Figure 5 shows tryptophol run in the same media as our sample. Our HPLC graphs do not have the peak at 162 m/z shown in the standard.

From the HPLC results, the modified bacteria do not show production of tryptophol. One potential explanation is that there is post-translational modification of these gene products. In particular, ARO10 has been shown to be expressed without exhibiting 2-oxo acid decarboxylase activity, indicating the presence of post-translational modification that may be present in yeast cells but absent in the modified E. coli cells.

In future experiments, PDC 1 could be used in place of ARO10 to produce the decarboxylase enzyme necessary for the degradation of tryptophan to tryptophol.