What is the estimate optimal of temperature? Hypothesis 3: How does the concentration of enzyme influence the rate of reaction? Why do you think this occurs? Materials Substrate solution Covet Spectrophotometer Timer Enzyme solutions Pipette for transferring fluids Thermometer Methods ; There is a choice of six different enzyme & substrate combinations in which the temperature will vary at which the reaction as well as the enzyme & substrate concentrations. L.A Substrate B/ Enzyme B was selected along with setting the wavelength to substrate vial/ 680 & the spectrophotometer to read light absorption.
. The temperature was set to 25 degrees & set to different experimental pH values starting at value 2 through value 10 (all even 3. Transferred 25 ml of the substrate B into the covet using the pipette. 4. Referring to the timer to determine When I minute has passed transferred 5 ml of enzyme 8 into the covet and quickly into the sample holder Of the spectrophotometer.
5. Recorded the absorption value at the 1 minute mark 6.Started a new experiment by removing the covet from the spectrophotometer and clicking the “New Experiment” button to peat and continue the steps for the remaining pH values. 7. Determined the estimate of optimal pH after the entire experiment for as a result. Temperature: 1.
Used the same Substrate/ Enzyme 8 & set the pH value to the optimal pH value – 4 from the pH results. 2. The temperature was set to 10 degrees & set to different experimental temperatures starting at value 10 through value 80 (all in the ass place) 3. Repeated the same steps as 4 – 7 as the pH experiment Reaction Rates: 1.Used the same Substrate/ Enzyme 8, set the pH value to the optimal pH value – 4 from the pH results, & set the temperature to the optimal temperature – 20 from the temperature results. 2.
Transferred 5 ml Of the enzyme a into the covet using the pipette. 3. Completed the same steps 4-7 as the pH experiment, but recorded my data based off of 10 second intervals through a total of 2 minutes to determine the absorption value. 4. After completing the steps transferring 5 ml of the enzyme a into the covet using the pipette, repeated the steps transferring 15 ml instead.