Ataa Amoah-Bediako

Mr. Kerr

SBI4U1-01

Thursday, Oct 20, 2022

Factors Affecting Catalase Enzyme Functioning using Hydrogen Peroxide

Purpose: The purpose of this lab/experiment was to observe the effects of different factors -

temperature, pH, enzyme concentration and substrate concentration - and the catalase enzyme

action conditions under those factors. With the use of hydrogen peroxide and the generation of

oxygen gas as an indicator of enzyme action.

Analysis Questions

1.
2. a) The general nature of the amino acid residue that would be found within the active site of

the catalase enzyme and facilitate the binding of the hydrogen peroxide molecule would be polar.

This is because of the difference in electronegativity between oxygen and hydrogen. Since

oxygen has an electronegative value of 3.44 and hydrogen’s value is 2.10, the difference between

them is 1.34. And according to the rules, since 1.34 is greater than 0.4 the molecule is polar with

oxygen being the delta-negative (higher electronegativity). Since the active site environment is

built to be an ideal environment for the enzyme, the active site would be polar. This is due to

polar molecules attracting other polar molecules (like likes like) because of the slight charges

they each have (slight electronegativity of the ends of the molecules). With this, the hydrogen

peroxide will be able to form strong dipole-dipole bonds with the active site. To summarize,

hydrogen peroxide is polar because the catalase enzyme (potato juice) is also polar.

b) One mechanism by which the interaction between hydrogen peroxide and the active site of

catalase might lead to the decomposition of hydrogen peroxide during the induced-fit process is

the third mechanism in which an enzyme can change the shape of a substrate. This mechanism

can distort, strain and weaken chemical bonds of the active site of the enzyme. Having there be a

decrease in the amount of energy needed to break the bonds. Having the catalase enzyme reduces

the energy needed to break the bonds of hydrogen peroxide, resulting in the decomposition of the

hydrogen peroxide.

3. a) For the chemical reaction shown the participation of catalase in the reaction should be

indicated with it being written on top of the arrows in the chemical equation shown.

b) Using the bond energy table, I determined the theoretical free energy change that would have

occurred during the reaction is -203kJ/mol. The total number of bonds broken is 4.

(calculation work shown on a separate sheet)

c) Assuming that all the energy conversions in the reaction are 100% efficient is a faulty

assumption because according to the rules (2nd law of thermodynamics), energy conversion can

never be 100% efficient, since some of the energy is always lost to the environment's increasing

entropy.

4. a) Hydronium ions become increasingly abundant in increasingly acidic conditions, and

Hydroxide ions become increasingly abundant in increasingly basic conditions (respectfully).

b) A tertiary bond might be disrupted by Hydronium and Hydroxyl ions' presence because ionic

tertiary bonds are created with positively charged R groups and negatively charged R groups.

And so since hydronium is a positively charged ion, it could ionize its hydrogen atom with a

negatively charged R group in an ionic bond, disrupting the ionic bonds. This disruption could

lead to the destruction (breaking of bonds) of the tertiary structure due to the negatively charged

R groups losing/weakening their attraction to the positive R group since it is receiving its wanted

positive charge from hydronium. Just like this but vice versa with Hydroxyl as it is a negatively

charged ion and would affect the ionic bonds by ionizing positively charged R groups.

5. a) 40℃ can be considered an outlier because looking at the data as the temperature increases

up to 15℃ the volume of gas(mL) does then decrease from 20℃ downward. But at 40℃ it

spikes to 6.4 mL and then continues to decrease.

b) When denaturation of the peroxidase enzyme occurred hydrophobic interactions (tertiary

bond), would have, most likely, been the first tertiary bond to break within the structure of the

peroxidase enzyme. This is because they have weak Van der Waals forces (London dispersion)

meaning that less energy would be needed to break the bonds. Therefore less heat would be used

for them to break, therefore the bonds would’ve broken first at a lower temperature. Whereas

ionic bonds would have, most likely, been the last tertiary bond to break, if we had higher
temperature trails within the lab. Since they are stronger bonds they’d need more heat energy to

break compared to the other bonds within the tertiary structure.

6. If I were to repeat the lab in order to generate more than the maximum 10.6mL of oxygen, I

would leave the reaction for longer than a minute (such as 3 minutes instead) because then the

remaining reactants would have more time to decompose and create more oxygen gas as the

change would help generate more than 10.6mL of oxygen gas.

7. As in different points of the experiment there existed different limiting factors in terms of

enzyme functioning, such as at ~80% substrate concentration a noticeable new limiting factor

occurs. Limiting factors are usually determined by the number of substrates at the beginning with

the increasing frequencies of collisions and reactions with the increased amount of substrates to

react with. But there can be a change in the liming factors that occur in the saturation level. This

change happens to despise the rate of reaction increases since once the enzyme molecules reach a

maximum rate, they combine with the substrate, which increases the substrate and reduces the

effect until there the rate of reaction lowers.

8. If I had been asked to identify a negative control trial in this experiment before being supplied

with the data and learning anything about potato peroxidase, I would have identified the

substrate at 0% trial to be negative control. This is because without hydrogen peroxide in the

reaction there wouldn’t be decomposition to produce oxygen gas proving that in trials in which

oxygen gas was generated, there had to be a decomposition reaction caused by hydrogen

peroxide. Since the breakage of hydrogen peroxide produced oxygen gas, water was created

along with oxygen gas. And so because of the enzyme, the reactions happen faster than without

it. Overall without the prior knowledge given to us, I would have identified substrate

concentration at 0% to be a negative control.