Using Cellobiase to Break Down Artificial Substrate, Mimicing Degradation of Cellobiose to Glucose for Creation of Ethanol

     For years, scientists have researched alternative sources of renewable energy that could eventually replace the dwindling supplies of oil and other natural resources used for feul. One field in which there is great promise is biofeuls, or feuls extracted from natural biomass. Certain biological oils and alcohol feuls such as ethanol could be the future because they have a cyclical, renewable energy process with little to no pollution. Therefore, we could potentially create an infinite supply of energy that is less toxic to the environment.
     To harness the energy in biological products, enzymes are necessary to break the mass down to its smallest components. Enzymes, usually proteins, bind to specific molecules called substrate and catalyze the reactions. Our lab will use the enzyme cellobiase, which simplifies the two-glucose molecule cellobiose to a single glucose molecule that can be harnessed for feul. Cellobiose is a simpler form of the sugar cellulose, which is found in the cell wall of plants, such as corn.
     As I have already said, scientists have obvious reasons for being interested in biofeuls. Glucose, specifically, can be converted to ethanol by fermentation. Ethanol is already being used as an energy source, but researchers are currently trying to perfect it. We are doing this lab to further our knowledge of enzymes and observe the degradation of cellobiose (artificial substrate) to glucose, which can be used to make ethanol.
     This lab is actually very simple. Rather than use actual cellobiose, which would break down into glucose that is invisible without a microscope, we will use an artificial substrate called p-Nitrophenyl glucopyranoside. We will add this substrate to a test tube with cellobiase, causing the substrate to break down into glucose and p-Nitrophenol.
     On the side, we will set up four beakers filled with a strong base. At different times, we will pour out some of our concoction from the test tube into each beaker. The base has two effects on our solution: first, it will kill the cellobiase, thereby ending the reaction immediately; also, the strong base will react with the p-Nitrophenol, turning it yellow. This allows us to have a visible indicator of how much glucose is being produced, and that the reaction was even successful at all.
     Because we are adding base at different times, we would expect the shade of yellow to be stronger the longer the reaction is occurring. The more the artificial substrate is broken down, the more glucose we will have. The variables in this lab are the different timepoints that we add the base, affecting the amount of reaction that can to occur. For our control, we will have one tube of artificial substrate in which we do not add cellobiase, so no reaction should occur.

Conclusion:
     Due to contamination by the previous period, our lab was unsuccessful. Our tube of substrate had been mixed with enzyme, so the reaction had already been completed before we began the experiment. All of our tubes showed the same dark shade of yellow because all the substrate had already been broken down. Also, our control sample was yellow even though we never added enzyme.