Enzyme Reaction Lab
Enzymes are proteins or nucleic acids that catalyze reactions. They are able to speed up reactions by reducing the activation energy of a reaction. Each kind of enzyme has a specific shape that matches its substrate so it can bind to its active site. Enzymes convert their substrates into a product. Enzyme activity are affected by factors such as temperature, pH, and time. If an enzyme is exposed to extreme heat, it will become denatured, that is, to become deformed and lose its original shape which causes it to be unable to bind with its substrate. This process is irreversible. Each enzyme also has an optimal pH value at which it
Hypothesis: Enzyme activity will increase as temperature increases until a certain point then enzyme activity will begin to decrease. Enzyme activity will also generally increase as time increases. …show more content…
Once the buffer had mixed with the substrate we removed 500 microliters of this new solution and added it to the "Start" cuvette. Using a clean tip, we pipetted 1 mL of enzyme into the 15 mL "Enzyme Reaction" conical tube, gently mixed, and then started our timer. We removed 500 microliters of the solution from the "Enzyme Reaction" tube and added it to the E1-E5 cuvettes at the corresponding times: 1 minute, 2 minutes, 4 minutes, 6 minutes, and 8 minutes. After these enzyme samples were collected, we used a clean pipet tip to remove 500 microliters of the solution from the "Control" reaction tube and added it to the "End" cuvette.
For the second portion of the lab, we labeled five cuvettes S1-S5 filled with 0, 12.5, 25, 50, and 100 nmol of p-Nitrophenol respectively. Then we used a calibrated spectrometer to record the absorbance of each of these cuvettes, the E1-E5 cuvettes from the first part of the experiment, and the "Start" and "End" cuvettes all at 410 nm wavelength and recorded our
Hypothesis: Enzyme activity will increase as temperature increases until a certain point then enzyme activity will begin to decrease. Enzyme activity will also generally increase as time increases. …show more content…
Once the buffer had mixed with the substrate we removed 500 microliters of this new solution and added it to the "Start" cuvette. Using a clean tip, we pipetted 1 mL of enzyme into the 15 mL "Enzyme Reaction" conical tube, gently mixed, and then started our timer. We removed 500 microliters of the solution from the "Enzyme Reaction" tube and added it to the E1-E5 cuvettes at the corresponding times: 1 minute, 2 minutes, 4 minutes, 6 minutes, and 8 minutes. After these enzyme samples were collected, we used a clean pipet tip to remove 500 microliters of the solution from the "Control" reaction tube and added it to the "End" cuvette.
For the second portion of the lab, we labeled five cuvettes S1-S5 filled with 0, 12.5, 25, 50, and 100 nmol of p-Nitrophenol respectively. Then we used a calibrated spectrometer to record the absorbance of each of these cuvettes, the E1-E5 cuvettes from the first part of the experiment, and the "Start" and "End" cuvettes all at 410 nm wavelength and recorded our