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Caden Reed

B3T

The Effect of pH on Foam Column Height in Various Solutions

Introduction

Enzymes are biological catalysts made of proteins that accelerate chemical reactions in

living organisms. Without enzymes, many of the essential biochemical processes that sustain life

would occur too slowly to maintain proper function. Each enzyme is highly specific to the

reactions it catalyzes, often working on just one type of molecule, known as the substrate.

Enzymes are crucial to processes like digestion, respiration, DNA replication. Enzymes work by

lowering the activation energy required for a chemical reaction to take place. Activation energy

is the minimum amount of energy needed to initiate a reaction. Enzymes achieve this by binding

to the substrate at a region called the active site. The active site is a specialized area on the

enzyme with a unique shape that matches the substrate, much like a lock fits a specific key. Once

the enzyme and substrate form an enzyme-substrate complex, the enzyme facilitates the reaction,

converting the substrate into products. After the reaction, the products are released, and the

enzyme can bind to another substrate molecule, repeating the process. Catalase is an enzyme

found in nearly all living organisms exposed to oxygen. Its primary function is to protect cells

from oxidative damage by breaking down hydrogen peroxide, a harmful byproduct of metabolic

processes, into water and oxygen. Without catalase, hydrogen peroxide would accumulate and

cause damage to cells, leading to oxidative stress. In a laboratory setting, catalase activity is

often measured by the amount of foam produced when it reacts with hydrogen peroxide, as

oxygen is released during the reaction.

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The activity of enzymes, including catalase, is highly sensitive to changes in pH. Each

enzyme has an optimal pH range where it functions most efficiently. Optimal pH can vary

depending on the organism and the specific environment where the enzyme is found. When the

pH is outside an enzyme’s optimal range, the enzyme’s structure can be altered. This happens

because pH affects the hydrogen bonds and ionic interactions that maintain the enzyme’s shape.

If the pH is too acidic or too basic, the enzyme may lose its shape and can no longer bind to the

substrate. As a result the enzyme can no longer do its job. In catalase in very low of very high pH

the enzyme’s ability to break down hydrogen peroxide is reduced. This is because the changes in

pH disrupt the active site’s ability to properly interact with the substrate. If the pH is normal it

will start to foam up. When the pH is to acidic of too basic more than likely they’re will probably

be no foam.In this experiment examining the effect of pH on catalase activity, the independent

variable is the pH level of the solutions in each test tube, while the dependent variable is the

foam column height, which indicates the level of catalase activity. To ensure a fair test, two

constants are the volume of each solution, which is kept at 1 mL, and the concentration of

catalase, standardized by adding the same amount (5 drops of 100% catalase) to each test tube.

This careful control allows for a clear assessment of how varying pH levels influence the activity

of the enzyme catalase.

My hypothesis is that the foam column height will vary depending on the pH of the

solution, with different pH levels either increasing or decreasing the activity of catalase in the

potato juice.This hypothesis predicts that the enzyme catalase will react differently in acidic,

neutral, and basic conditions, which will affect the amount of foam produced. The Null

hypothesis is that the pH of the solution has no effect on the foam column height, meaning the

activity of catalase remains constant across all pH levels. The Alternative Hypothesis is that the
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pH of the solution significantly affects the foam column height, meaning the activity of catalase

varies depending on the pH level of the solution.

Experimental Design

First thing we did befor even waling in to the lab was make sure we had the

appropriate apparel to walk into the lab. Then we put on our labcoats so that no chemical can get

on us, and then we sat down at our asinged seats as we normally do until or instructor tells us we

can start the lab. The lab normally begins around 5:00pm. When our instructor tells us to do so

we gab our gloves and goggles. Then head over to the pH station which is on the counter next to

the sink. The materials in the dest where vinegar, RO water, 1M NaHCO3, 2M NaOH, potato

juce, 3% H2O2, pH strip tester, transfer pipette, ruler, water wash bottle and 4 beakers labled

A,B,C,and D. Once we have read our lab manual. Then we tested the pH for vinegar, Ro water,

1M NaHCO3, and 2M NaOH with the pH strips we where given. Then we when to the first step

which was add 1mL vinegar to tube A. 2ed step was to add RO water to tube B. Then to tube C

add 1mL of 1 M NaHCO3 (sodium bicarbonate). Then add 1mL of 2 M NaOH (sodium

hydroxide) to tube D. Then add 5 drops of your 100% potato juice to all of the tubes and mix

them gently. We waited around 1 minute after mixing the products. Then added 1mL of 3%

H2O2 to all of the tubes and agin gently mixed them. Then we waited around for 2 minutes.

Then we measured all of the tubes foam hight with a ruler with the cm of course. We did this by

puting the ruler right above the liquid inside of the tube where the foam was just starting. Then

we made sure that the ruler was exactly on the staring line of inbetwean the liquid and foam.

Then we did that for all of the tubes and recorded or findings in our lab manual. Then we washed

out all the beakers out with the spesial reinsher we were giving by our instructor, and made sure

that there was no more foam left for the other grupes behind us. (Lab Manual,2019)
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Result Summmory

In this experiment, we tested the effect of different pH levels on catalase activity

by measuring the height of the foam column produced. In Tube A, with vinegar (pH 3), the foam

column height was 0.1 cm, indicating low catalase activity in an acidic environment. In Tube B,

with RO water (neutral pH 7), the foam column height was 2.5 cm, suggesting higher catalase

activity at a neutral pH. Tube C, containing a weak base 1 M NaHCO₃, (pH 10), it produced a

foam height of 1 cm, indicating moderate enzyme activity in a slightly basic environment.

Finally, in Tube D, with a strong base 2 M NaOH, (pH 14), and no foam formed 0 cm.

Test Tube Actual pH Foam Column Other Groups Average

Height (cm) Height (cm) (cm)

Tube A 3pH .1cm 0cm .05cm

Tube B 7pH 2.5cm 2cm 2.25cm

Tube C 10pH 1cm 1.7cm 1.35cm

Tube D 14pH 0cm 0cm 0cm

Table 1: Raw data of foam hight contradicting to different pH levels

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Graf 1: Foam hight with different pH conditions

Conclusion

The conclusion of this experiment is that catalase enzyme activity is affected by pH, with

the enzyme showing the highest activity at a neutral pH and reduced activity in highly acidic or

basic conditions. Explanation: Catalase is an enzyme that breaks down hydrogen peroxide into

water and oxygen, producing foam as a byproduct in the presence of potato juice. Enzymes,

including catalase, have an optimal pH range where they function most effectively. Neutral pH

and slightly basic seamed to be the best by not changing the shap of the enzymes to much.Unlike
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in highly acidic and strong basic pH where the foam was much lower indicating that catalase

activity is significantly reduced in acidic conditions. Acidic pH can disrupt hydrogen bonds and

other interactions within the enzyme, causing it to lose its functional shape or denature, which

reduces its ability to catalyze the reaction. In conclusion this activity is best at a neutral pH and

shows a decreases in both acidic and highly basic environments. This trend suggests that extreme

pH levels disrupt the enzyme’s structure there for inhibiting its function.

I like this experiment because it was fun to actually see what was happening in real time. I would

still change how we did things because we use the pH strips to find out the pH levels, but the pH

levels were already given to us. So we wasted some time. I would try to make sure the constant

or as consistent and try to make sure everything was the same except for the pH. I would

probably make sure we had our measurements right because I believe there is some confusion in

counting the drops.

References

Life: The Science of Biology, 12th ed (2020). Hillis, Heller, Hacker, Hall, Laskowski, and

Sadava. Sinauer Associates. (ISBN 9781319440985). Achieve is required for this

course.

Biology 1406 Laboratory Manual 2nd Edition, Hayden-McNeil. (ISBN 9781533937612)