Purpose: The purpose of this project is to find out the effect of the enzyme activity by looking at enzyme concentration, substrate concentration, and the effect of pH and temperature on enzyme activity.
Hypothesis: If procedures are followed correctly then the effect on enzyme activity will be found.
Hydrogen Peroxide 3% and 1.0 %
Filter Paper Discs
Stopwatch or Timer
1. Gather all materials
2. Set up 5 vials containing 40ml of 4 % hydrogen peroxide in each. Measure and record the depth of the hydrogen peroxide in the vials.
3. Dilute the enzyme as follows. Make each dilution in a separate cup.
100 units/ml = 12 ml 100 units/ml + 3 ml cold dH2O
80 units/ml = 12 ml 100 units/ml + 10 ml cold dH2O
50 units/ml = 3 ml 100 units/ml + 12 ml cold dH2O
20 units/ml = 3 ml 100 units/ml + 12 ml cold dH2O
0 units/ml = 20 ml cold dH2O
4. Using the forceps, dip a filter into the enzyme solution at 100 units/ml, then remove it and drain it on a paper towel. Drop the disc into the vial of hydrogen peroxide labeled 100 units/ml and time how long it takes the filter to rise to the surface. Repeat this procedure for each of the other enzyme dilutions. Record result.
5. Obtain 1 vial of catalase at 100 units/ml.
6. Dilute the substrate (hydrogen peroxide) as described below.
3.0% h2o2:40 ml 3% h2o2
1.5% h2o2:20ml 3% h2o2 + 20ml distilled water
0.75% h2o2 :10 ml 3% h2o2 + 30 distilled water
0.38% h2o2 : 5 ml 3% h2o2 +35 ml distilled water
0.0% h2o2 : 40 ml distilled water
7. Dip a filter into the catalase, drain on a paper towel and then drop the filter into the 3% H2O2. Time how long it takes the filter to rise to the top. Repeat this procedure for each of the substrate dilutions. Record your results in the appropriate data chart.
8 Obtain 1 vial of 40 ml 1% h2o2. Measure and record the depth of the hydrogen peroxide.