SOUNDARYA DASARAMOOOLE 09/03/23

Soundarya Dasaramoole 09/03/23

The Effect of Enzyme Concentration on Enzyme Action

Aim: To know which catalase enzyme concentration when mixed with H2O2 increases the
rate of reaction.

Background Introduction

A catalyst speeds up the rate of a chemical reaction without being used up.
An enzyme is made up of proteins that also act as biological catalysts it contains amino
acids linked with one or more polypeptide chains.

Catalase can be found in all main production sites in H2O2 (hydrogen peroxide)

In a synthesis reaction when hydrogen is reacted with oxygen water is formed.

word equation: 2H2 + O2 ——> 2H2O
All synthesis reactions are reversible reactions.

The gas produced in this experiment is oxygen. To know if oxygen is present in a reaction
a glowing splint near oxygen gas should be able to relight the splint.

Catalysts increase the rate of reaction by lowering the activation energy which helps
increase the frequency of successful collisions of the reactants.

Equipment and Chemicals

 8 x 10 ml Syringe
 5 x 50ml Beakers
 5 x Test tube
 1 x Test tube rack
 5 x 100ml Measuring cylinder.
 1 x 100ml Enzyme Catalase solution (4000 EU)
 1 x 100ml H2O2 solution
 1 x Water bath temperature set to 30 degrees Celsius.
 1 x stopwatch
 1 x 3ml detergent solution
 1 x marker pen
 1 x 200ml Deionised water
 1 x 5ml Pipette
SOUNDARYA DASARAMOOOLE 09/03/23

Method

1. Wear all the safety equipment before starting any procedure like wearing a lab coat,
glasses, gloves and hair tied.
2. Safely organise all the equipment needed on a clean table.
3. To prepare the different concentrations of the enzyme catalase solution from
4000eu - 500eu it needs to be diluted with water (deionised).
4. Pour the 4000eu catalase solution into beaker A with no added water.
5. Take 5 ml of catalase solution beaker A using a 10 ml syringe and pour it into a
different beaker (labelled B) and using a 10 ml syringe measure water up to 15 ml
then pour this water into the beaker labelled B and mix well. (3000eu)
6. Take 5ml of the 4000eu catalase solution using the 10ml syringe and pour it into a
different beaker labelled number C. Measure 10ml of water using a 10ml syringe
then pour it into the beaker C and mix well. (2000eu)
7. Repeat method 6 for beakers D (1000eu) and E (500eu)
8. Have 5 x measuring cylinders and label them from 1-5. Then take the hydrogen
peroxide solution and pour it into a 50ml beaker using a clean 10ml syringe pour 5ml
of that hydrogen peroxide solution into each of the measuring cylinders labelled
from 1-5.
9. Then add 3ml of detergent into each of the measuring cylinders labelled 1-5 and
swirl to mix well.
10. Keep the water bath at 30 degrees Celsius ready before doing all of the above.
11. Take 5 x test tubes place them into a test tube rack and label it from A. Take the
catalase solution of all the concentrations made and using a 5ml syringe take 2ml of
catalase solution and pour it into the test tube matching label from the beaker to
tube A-E.
12. Then carefully place all the test tubes with catalase solution and the measuring
cylinder with the hydrogen peroxide with the help of the rack into the water bath for
5 minutes (pre-incubation).
13. After 5 minutes carefully add the different concentrations of enzyme catalase
solution into the measuring cylinders A to cylinders 1-5 respectively at timed
intervals of 20 seconds (staggered start).
14. Take out all of the test tubes and place them on a table carefully.
15. Allow the enzymes and substrate (hydrogen peroxide) to mix well for 2 minutes.
16. After 2 minutes carefully take out all of the measuring cylinders place them on a
table and measure the height of the froth from 1-5 at timed intervals of 20 seconds.
17. Repeat methods 17 and 18 once.
18. Calculate the average height of the froth for each enzyme concentration.
SOUNDARYA DASARAMOOOLE 09/03/23

Raw data and observation

Catalase concentration (EU) Height of the froth (ml)

4000 36

3000 32

2000 15

1000 14
500 12
SOUNDARYA DASARAMOOOLE 09/03/23

Table of class results with average

Catalase solution Height of Froth (ml)

(EU)
Group 1 Group 2 Group 3 Group 4 Group 5 Average
4000 47 43 65 37 36 45.6
3000 30 38 50 34 32 36.8
2000 15 12 17 15.5 15 14.9
1000 12 13 8.5 14 14 12.3
500 10 10 7 13 12 10.4
SOUNDARYA DASARAMOOOLE 09/03/23

Conclusion

As per aim, when enzyme catalase mixed with H2O2 and detergent solution the catalase
concentration of 4000 EU gives out more products and increases the rate of the reaction
with average froth height of 45.6ml.
In conclusion, as the concentration increases the height of enzyme catalase froth increases.

Evaluation

When using a syringe to add catalase solution it forms bubbles and disrupts volume
accuracy. To avoid this measuring cylinder can be used to be more accurate in volume
hence having more accurate results.

Can also use a pipette to be more accurate when diluting catalase solution to make different
enzyme concentrations.

Repeating the procedure three times to have a better training and understanding of the
experiment and give out and show reliable results.
SOUNDARYA DASARAMOOOLE 09/03/23
SOUNDARYA DASARAMOOOLE 09/03/23