St.

Marys Grammar School, Magherafelt

GCSE Biology Coursework

The Effect of Substrate Concentration on the Rate of an Enzyme-Controlled Reaction
The Reaction;
Hydrogen peroxide (H2O2) is formed in body cells constantly. It is broken down to water and oxygen. This reaction is catalysed by the enzyme catalase. Catalase is found in most tissues, including potato.

4H2O2
catalase

4H20 + 2O2

Name: Dermot McGuckin Candidate No: ______________________________ Centre No: _________________________________ Date: ______________________________________

(2009)

St.Marys Grammar School, Magherafelt

PLANNING (a) Background Knowledge:

What is an enzyme? An enzyme is a biological catalyst that speeds up the rate of chemical reaction. It is made of protein. Explain how enzymes work. Include a labelled diagram. An enzyme reacts with a substrate. The enzyme must be a complimentary shape to the active site. The lock and key hypothesis is when the enzyme and the active sit of the substrate are specific or complimentary to each other.

List the factors that affect the rate at which enzymes work and briefly explain each factor. Temperature: As temperature increases the rate of reaction increases. This is because the enzyme and substrate have more kinetic energy which means more successful collisions. There is more H202 breakdown to give more o2 and h2O. When the temperature is above 40 degrees the enzyme becomes denatured and the rate of reaction decreases.

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St.Marys Grammar School, Magherafelt

PH: Enzymes like to work in a specific range. When an enzyme is out of its range it becomes denatured. The rate of reactions therefore decreases.

Enzyme concentration: As enzyme concentration increases the rate of reaction increases, because there are more active sites available therefore more successful collisions. Shown by part A on graph. At B it levels of due to limiting factors such as temperature substrate concentration and PH.

Substrate Concentration: As substrate concentration increases the rate of reaction increases also because there are more substrate molecules and H202 to react with the enzyme molecules and therefore more successful collisions. At part B of the graph it starts to level of due to limiting factors such as: enzyme concentration, PH and temperature.

(b)(i) Prediction:

P6a (iii)

As we increase thee substrate concentration the rate of reaction will increase. Therefore, there will be an increase in volume of oxygen produced per unit time. However, the volume of the oxygen will level off at a certain substrate concentration due to limiting factors such as: Enzyme concentration, PH and temperature. (2009)

St.Marys Grammar School, Magherafelt

(ii) Explanation of Prediction;

P8a (iii)

As you increase the substrate concentration of the hydrogen peroxide, there will be more hydrogen peroxide molecules. There will therefore be more successful collisions between the H2O2 and the catalyse, therefore the volume of ozygen will increase because of more O2 breakdown realising more O2, However the graph will level off due to limiting factors.

(c) Variables: Independent Variable:

(i) What will you change? I will change the substrate concentration (H2O2)

P6 (a) (ii)

(ii) Range:
How many different substrate concentrations will you use? I will use five different substrate concentrations. What range of substrate concentrations will you use? 20% 40% 60% 80% 100% Draw a table to show how you will achieve each concentration.
Concentration of H2O2 20 40 60 80 100 Volume of H2O2/Cm3 Volume of Water/Cm3

P6b

4 8
12

16 20

16 12 8 4 0

20 20 20 20 20

Dependent Variable:
What will you measure to get results? I will measure the volume of oxygen produced per 2minutes/cm3

P6 (a) (ii)

Controlled Variables:
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P6 (a) (ii)

St.Marys Grammar School, Magherafelt List all the factors that must be kept the same to make this a fair test and explain how you will control each one. Enzyme Concentration: I am going to use a cork borer to cut the potato cylinders, and then use a 30cm ruler to ensure the potato cylinders are all the same length of 3 cm3. We will use the same type of potato also. Volume of Solution: I will ensure with the aid of a 25 cm3 measuring cylinder that the same volume of solution is added to each boiling tube. Temperature: We will keep all boiling tubes in the same room which will ensure they are all at room temperature 21oC. We will ensure this by using a thermometer. PH: To ensure the PH remains the same throughout we will not add any acid or alkali solution to the boiling tube. Time: All observations will be noted during a 2 minute time period, we will use a stopwatch to ensure we do not go overboard.

(d) Method:

P2a P4a/b P6 (a) (i) P8 (a) (i) (i) Give step-by-step instructions of your method, including a labelled diagram of your apparatus. Collect apparatus and set up as the diagram shown. Put on the safety goggles to prevent chemicals getting in your eyes. Fill your trough full with water. Using a cork borer size 5, produce 5 potato cylinders for each concentration of hydrogen peroxide. Using a 30cm ruler ensure all cylinders are 3cm and take of any remaining skin on the cylinders, use a scalpel.

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St.Marys Grammar School, Magherafelt Using the 25cm3 measuring cylinder measure out your 5 concentrations of your hydrogen peroxide( see earlier table eg: to make 20% hydrogen peroxide concentration add 4 cm3 of hydrogen peroxide to 16cm3 of water Add all concentrations to separate boiling tubes and label them. Take the 1cm3 measuring cylinder. Place in the trough completely filled with water an invert. Take one of the potato cylinders and add to your 2% hydrogen peroxide solution then attach the bunged delivery tube to the boiling tube. Take the delivery tube and insert it under the 10cm3 measuring cylinder Start the stopwatch as soon as you the first oxygen bubble being released. Time for 2 minutes. After 2 minutes record the volume of oxygen produced per 2mins/cm3 by recording how much water has been displaced from the 10cm3 measuring cylinder. E.g.: If the measuring cylinder starts with 10cm3 of hydrogen peroxide and ends with 8cm3 of water the volume of oxygen produced equals 2cm3. Repeat the above steps for all 5 different concentrations Repeat the whole experiment for all the concentrations 3 times and take an average to increase reliability. Record results in a suitable table.

(ii) Precision List all the measuring equipment that you will use, including sizes. (2009) A 30cm3 Ruler Thermometer 10cm3 measuring cylinder 25cm3 measuring cylinder Stopwatch

P8 (a) (ii)

St.Marys Grammar School, Magherafelt Cork borer size 5

(iii) Reliability: How will you increase the reliability of your results?

P8 (a) (ii)

To increase reliability I will repeat the experiment three times and take an average for each concentration.

(e) Results: P8b (i) Draw the blank table that you will put your results into. Include all columns, column headings and units.

Concentration Volume of O2 produced per of H2O2 % 2mins/cm3 1 2 3 Average 20 40 60 80 100

(ii) Sketch and label the axes of the graph that you would use to plot your results.

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St.Marys Grammar School, Magherafelt

Y Average Volume of O2 produced per 2mins/cm3

X Concentration of Hydrogen peroxide/%

INSERT TABLE OF RESULTS

O2a, O4a, O4b, O6a, O6b, O8a

Concentration Volume of O2 produced per of H2O2 % 2mins/cm3

20 40 60 80 100

1 1 1.8 2.4 3 3

2 0.8 1 1.8 2.4 3.6

3 1 1.5 3 3.5 7

Average 0.9333 1.4333 2.4 2.97 4.5333

INSERT GRAPH

I4a, I6a

INTERPRETATION Interpretation: Trend:

Describe the trend shown by your graph. My graph shows that as the concentration of hydrogen peroxide increases the vole of oxygen also increases. For example: 20% of hydrogen peroxide concentration I2a I4b

(2009)

St.Marys Grammar School, Magherafelt released 1.2cm3 of O2 and when the concentration was increased to 60% the amount of oxygen released also increased to the figure of 3.4cm3. However my graph did not level off as I predicted.

Conclusion:

I6b I8a (i) Use your background knowledge and the explanation of your prediction to explain the trend that you identified above. When you increase the substrate concentration the hydrogen peroxide molecules increase. This will therefore increase the number of successful collisions between the enzyme and the substrate and in turn this increased enzyme/substrate complexes. This increases the volume of oxygen released , in other words increase the rate of reaction. However the graph did not level off which means temperature, pH and enzymes concentration were not limiting factors.

(ii) How well do your results support your prediction?

I8b

The graph didnt level off as I predicted but as we can see the substrate concentration increased so the rate of reaction therefore increased as well as predicted.

Evaluation: Of Method:
Suggest how your method could be improved

I6c

(2009)

St.Marys Grammar School, Magherafelt To improve my method I could do the following:

Replace 10cm3 measuring cylinder with a gas syringe. Which will make the collection of oxygen more efficient. Use clear tubing. This will allow anyone to see if there are any blockages. Repeat the experiment more than 3 times and find a more accurate average. Use a beehive shelf to help collect the oxygen more efficiently. A greater range of percentage concentrations, so the trend can be more accurate.

Of strange (anomalous) results:

(i) Identify any anomalous results. 20% Concentration 80% Concentration

I8(c) (ii)

(ii)

Suggest the possible cause(s) of these anomalous results.

An accidental increase in temperature could be the cause for the anomalous results as the higher the temperature the quicker the rate of reaction. There could of been inconsistency in the amount of catalyse in the potato cylinders. There also could of been a blockage in the delivery tube , this would present any oxygen from entering. There could of also been human error with the regards of the size of the potato cylinders.

Of Reliability of Results:
(i) Why do you think your results are reliable?

I8(c) (i)

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St.Marys Grammar School, Magherafelt I believe my results are reliable as I repeated the experiment 3 times to increase the reliability of my results.

(iii)

Could you make them any more reliable?

Yes, I could of used a greater range on concentrations to increase reliability and I also could of completed the experiment more than 3 times and took an average to increase reliability.

(iii) Based on your answer in (i), state whether you think that your evidence is good enough for a firm conclusion. I8(c) (ii)

I believe my evidence is good enough for a firm conclusion as it has been carefully planned and well researched. I repeated the experiment on three occasions to increase reliability and they showed that my results followed my prediction.

(2009)