Abstract
P450 is involved in so many processes due to the various chemically relevant reactions which it efficiently catalyzes, the most striking of which is its ability to catalyze C-H hydroxylations. As anyone with organic chemistry experience knows, this is a very difficult reaction from which to achieve product turnover. This is because it requires a number of harsh reagents (heavy metals) and often times arduous reaction conditions as well. In order to emulate this process in-vitro and achieve product turnover we must consider a number of potential issues and challenges. First we must immobilize the protein on a surface, preferably an electrode. Second, we must ensure the presence of electron transfer into the heme center; this would emulate the NADPH in the native enzymatic cycle. Thirdly, we must ensure the enzyme?s reaction with dioxygen. All three of these goals must be accomplished without perturbing the structure of the protein too extensively or we will face the issue of forming hydrogen peroxide, as aforementioned. The overall goal is to be able to link the protein to an electrode and feed discrete into the heme so that it can undergo the diagramed cycle in-vitro. This will allow us to achieve the active form of the enzyme and perform the desired chemistry on the substrate.
