| Category | CELL | L15 | Testing the Geometrical and Chemical Structure of |
| Methylbutenol Synthase: Find |
| Abstract | The purpose of this experiment is to see if changing the chemical |
| structure of Methylbutenol (MBO) synthase’s active site from a |
| Phenylalanine to a Tyrosine will effect the production of MBO and |
| isoprene. By finding alternative ways to increase the production of MBO |
| and isoprene, it will allow for an increase in rubber production and |
| various uses in biotechnology. It was hypothesized that with the |
| chemical and geometrical change, a greater production of MBO and |
| isoprene will be produced because Tyrosine is polar due to the |
| hydroxide group. The experiment was conducted by using proteins from |
| last year, which went through a site directed mutagenesis. Next, the |
| proteins underwent a protein expression was applied by suspending |
| bacterial cells, vortexing, and then centrifuging the bacterial cells. The |
| supernatant was run through a western blot. A functional assay was |
| used to color the gel in order to see the bands of protein. Gas was then |
| extracted from the protein and cycled through a gas chromatograph. |
| The results showed that there was little change in the amount of MBO |
| and isoprene production. The ratio had a slight shift toward the |
| production of isoprene. This shows that the geometry and the chemistry |
| had an effect on MBO synthase. |
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