Running head: DIFFERENCES OF SUBSTRATE CONCENTRATIONS

Differences in the Concentrations of Hydrogen Peroxide in Consistent Substrate Concentrations

Naomi Belcher, Julia Calhoun, Rayvin Ewers, and Katarina Mayer
Owensboro Community and Technical College
January 21, 2016

DIFFERENCES OF SUBSTRATE CONCENTRATIONS

Abstract
This experiment was posed to find that if a higher concentration of substrate was added to
a solution with a constant concentration of enzyme. In this case catalase was the enzyme and
hydrogen peroxide (H2O2) was the substrate. To do this, water (H2O) and different
concentrations of H2O2 were put into four test tubes and a drop of the enzyme catalase was
added and then the rate of reaction was measured by a pressure sensor. As the concentration of
the H2O2 increased, the slope of each line was supposed to increase, but technological
difficulties prevented this from being collected. Barring complications, this experiment turned
out to prove our hypothesis correct because when the higher concentration of H2O2 was added,
the rate of reaction increased.

DIFFERENCES OF SUBSTRATE CONCENTRATIONS

Background
First, the in class experiment showed the median of the reaction. In this we used 3
milligrams of H2O2. So when we used 1.5 milligrams and 6 milligrams of H2O2, it gave more
of a representation to predict what the lines on the graph would be at 1.5 and 6 ml.
Researching the experiment we found that, the reaction begins very swiftly. As soon as
the enzyme and substrate are mixed, bubbles of oxygen are released quickly. A large volume of
oxygen is collected in the first minute of the reaction. As the reaction continues, however, the
rate at which oxygen is released gradually slows down. The reaction gets slower and slower,
until it eventually stops completely.
Hypothesis
When there is a higher amount of substrate concentration, the rate of reactions will be
higher than the reactions that took place with a lower amount of substrate concentration.
Materials and Methods
Materials include: four test tubes, 5 beakers, 3 different concentrations of H2O2 (in this
experiment 1.5%, 3% and 6% were used), H2O, safety equipment, pipettes, Vernier gas pressure
sensor, Logger Pro software, Vernier LabQuest Mini, test tube stand, a computer, labeling
equipment, and catalase concentration of 200 units/mL.
To replicate this experiment, follow these steps:
1. Put on safety equipment
2. Gather all the materials that are listed above
3. Hook up the Vernier LabQuest Mini and the Vernier Gas Pressure Sensor together and
then up to the computer that has the LoggerPro software downloaded and pulled up

DIFFERENCES OF SUBSTRATE CONCENTRATIONS

4. Put all four test tubes in the test tube stand and label them 1, 2, 3, and 4
5. Put all solutions (H2O, all concentrations of H2O2, and catalase) in separate beakers
6. Add 3 mL of H2O to each of the 4 test tubes in the rack
7. Add 3 mL of 1.5% H2O2 solution to test tube 2
8. Add 3 mL of 3% H2O2 solution to test tube 3
9. Add 3 mL of 6% H2O2 solution to test tube 4
10. Add one drop of catalase solution to test tube 1 and put the stopper into the test tube and
hit collect on the Logger Pro Software
11. Collect the data for 200 seconds while agitating the tube during collection
12. Repeat steps 10 and 11 for test tubes 2, 3, and 4
Results
The result of our experiment is that the concentration of the substrate does affect the
oxygen that is produced. Our substrate was H2O2, we used the concentrations of 6%, 3%, and
1.5%. The highest concentration, 6%, had the highest rate of kPa/s produced. The lowest
concentration, 1.5%, had the lowest rate of kPa/s produced. All are shown in Figure 1 and 2. The
higher the concentration of H2O2, the steeper the slope of the graph should have been.

DIFFERENCES OF SUBSTRATE CONCENTRATIONS

Figure 1

Figure 2
Discussion
The results of the experiment coincide with the hypothesis. We predicted that the more
catalyze, the faster the enzymatic reaction. Also we observed downfalls in the experiment: our
gas pressure sensor didnt work accurately, the Logger Pro stopped working, the computer died,
and the catalase aged. Even with mix of all the disruptive occurrences, the reading still came out
positively. The improvements that could be made are check all the equipment before and make
sure they work before starting, do all runs of the experiment in one day so the catalase doesnt go
bad, and go through more experimental runs for more accurate results.
Conclusion

DIFFERENCES OF SUBSTRATE CONCENTRATIONS

In this experiment we have learned that the substrate concentration does effect the
oxygen produced by the catalase. Catalase is an enzyme found in living organisms that changes
hydrogen peroxide (H2O2) into water and oxygen. When the substrate concentration was at the
highest of 6%, more oxygen was produced by the catalase. Less oxygen was produced when the
lowest substrate concentration of 1.5% was used. So, as the concentration of the substrate
increases so does the enzymatic reaction.

DIFFERENCES OF SUBSTRATE CONCENTRATIONS

Works Cited
The effect of substrate concentration on enzyme activity. (n.d.). Retrieved January 21, 2016,
from http://www.ucl.ac.uk/~ucbcdab/enzass/substrate.htm