Experimental Design Graphic Organizer

Goal of the investigation

RESEARCH QUESTION: What are you testing?
How does pH value affect the rate of reaction of enzyme?
INDEPENDENT VARIABLE: The thing you plan to change/manipulate DEPENDENT VARIABLE: The thing you plan to measure
The pH of the environment where enzymes are located. The time required for the enzyme spheres to sink and then float up to
the surface after being put into the hydrogen peroxide solution.
HYPOTHESIS: predicted relationship between your independent and dependent variable
(Example: An increase/decrease in IV will cause an increase/decrease in DV)
When the pH is between 2-6, at pH=6, the time required for enzyme spheres to sink and float up to the surface of H2O2 is the least. A
decrease in pH results in more time needed for completing this process.
Validity of the experiment
CONSTANTS: all the things that must be the same for all treatments # OF TRIALS: How many times will you repeat your experiment?
The temperature of the environment (23 Celsius). We’ll repeat the experiment 5 times for each trail.
The amount of yeast in each group (5 spheres).
The shape and size of each yeast sphere.
The amount of hydrogen peroxide in each group (10ml).
CONTROL GROUP(S): The baseline that you compare everything to EXPERIMENTAL GROUP: The trial in which you manipulate the IV
Yeast spheres with the same shape and size will be divided into five
NEGATIVE CONTROL: No treatment applied & no effect expected groups of five. The groups will then be put into 10ml hydrogen
There will be five spheres that have the same shape and size as those peroxide with a pH of 2, 3, 4, 5, and 6, respectively.
in the other five groups but contain no yeast. Each of them will be
placed and treated with an experimental group.

POSITIVE CONTROL: Known treatment will well understood effects

None.
Procedure
MATERIALS: What materials will you need? METHOD: What are you going to do?
10% yeast solution 1. Add 20ml 10% yeast (Saccharomyces cerevisiae) to 20ml 2%
2% sodium alginate solution sodium alginate solution. Mix well with a glass rod.
0.15M CaCl2 solution 2. Draw up the yeast-sodium alginate solution into a 30ml syringe.
HCl solution Carefully wipe off excessive liquid from the syringe tip.
NaOH solution 3. Very slowly depress the plunger so that drops of the yeast-sodium
50ml 0.6% hydrogen peroxide solution alginate solution fall into the beaker containing 50ml 0.15M CaCl2.
Tap water Continue releasing the rest drops into the beaker. Once the drops
Distilled water float, dispose them.
Eye droppers 4. Obtain a small strainer and hold it over a clean beaker. Carefully
Glass pod (1×) pour the contents of the beaker containing the yeast-sodium alginate
10ml graduated cylinder (1×) spheres into the strainer.
30ml syringe(1×) 5. Once drained, carefully rinse the spheres under a slow running
Beaker faucet, using the tap water. Then put the spheres into empty beakers
Forceps (1×) for further use. Prevent them from drying.
Strainer (1×) 6. Add 8ml water into 2ml 3% hydrogen peroxide to make 10ml 0.6%
pH meter (1×) H2O2 solution with a graduated cylinder. Pour the diluted solution
Timer (1×) into a test tube. Repeat the step for the other four test tubes.
7. Add HCl and NaOH slowly to each test tube using eye droppers
while having a pH meter to monitor the pH of the solution
simultaneously until the value reaches 2, 3, 4, 5, and 6, respectively.
8. Using forceps, remove one yeast sphere from the beaker and drop
it into a test tube. As soon as the sphere touches the surface of the
hydrogen peroxide and begin to sink, start timing. Keep timing until
the sphere reaches the surface again. Record the timing result in a
table. Repeat the step for five more times in the test tube because
one group contains five yeast spheres (experimental group) and one
sphere that that has the same shape and size as those in the other
five groups but contain no yeast (negative control).
9.Repeat step 8 for four more times for each pH values respectively.
Record all the timing results in the table.
Predicted Results
PREDICTED RESULTS: Based on your hypothesis and proposed procedure, what experimental results would confirm your hypothesis?
There is an optimum pH range, in which peroxidase has the fastest rate of reaction.
As the pH of hydrogen peroxide solution decreases below the optimum, it takes more time for a peroxidase sphere to float up to the surface of
the solution. As the pH of hydrogen peroxide solution increases below the optimum, the time required for a peroxidase sphere to float up to
the surface of the solution increases as well.

After recording the data and drawing a graph showing the trend, as pH increases, the time of peroxidase sphere floating up to the surface
decreases, reaches the lowest value when pH is neutral, and then increases.
References:

Battcock, M., & Azam-Ali, S. (1998). FERMENTED FRUTIS AND VEGETABLES. A GLOBAL PERSPECTIVE. Rome FAO Italy IT.
https://www.fao.org/3/x0560e/x0560e08.htm

Buckley, B. G., By: & Buckley, G. (2020, August 28). Dumbo Octopus - facts and beyond. Biology Dictionary. Retrieved December 1, 2021,
from https://biologydictionary.net/dumbo-octopus/
Hydrochloric Acid Calculate pH Values of Hydrochloric Acid Solutions. (2018). OxyChem.
https://www.oxy.com/globalassets/documents/chemicals/products/chlor-alkali/tech-calculated_ph_values_HCl.pdf

Milne’s Science Tech. (2021, Feburary 13). How pH effects enzyme activity [Video]. YouTube. https://www.youtube.com/watch?
v=34VkaCJggOM

Narendranath, N. V., & Power, R. (2005). Relationship between pH and Medium Dissolved Solids in Terms of Growth and Metabolism of
Lactobacilli and Saccharomyces cerevisiae during Ethanol Production. Applied and Environmental Microbiology, 71(5), 2239–2243.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1087585/

Norham High School Science. (2017, September 30). pH and Enzyme Activity [Video]. YouTube. https://www.youtube.com/watch?
v=_wkuUwA4bdA