ALWADI INTERNATIONAL SCHOOL BIOLOGY GRADE 9 5.

ENZYMES NOTES

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❖ In living cells, thousands of different chemical reactions called metabolic reactions

happen every second to enable the cell to function. For example, in liver cells
chemical reactions break down the toxic substances into harmless products. If
these reactions occur slowly then wastes can build up and poison the cell.
❖ The rate of a chemical reaction can be increased in the laboratory by increasing
the temperature. This gives more molecules sufficient energy to react with each
other. However, in the body, higher temperatures cannot be used to speed up
reactions as this would damage cell proteins and prevent them from functioning.
❖ Living cells solve this problem by producing proteins called enzymes.
❖ Enzymes are specialized protein molecules facilitating most of the body’s metabolic
processes – such as, supplying energy & digesting food. Enzymes are vital to our
health and change the rate at which chemical reactions happen, without any
external energy source added or by being changed.
❖ The temperature inside organisms is low (e.g. the human body temperature is about
37 °C) and without catalysts, most of the reactions that happen in cells would be
far too slow to allow life to go on. Only when enzymes are present to speed them
up, the reactions will take place quickly enough.
❖ Enzymes are biological catalysts, which are made up of proteins, and they speed
up chemical reactions.

3D computer model of an enzyme Enzymes as represented in drawings

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PROPERTIES OF ENZYMES

1. Enzymes are defined as biological catalysts which speed up the rate of the reaction
without being changed at the end of the reaction.
2. Enzymes are made up of proteins, having
a specific shape.
3. Enzymes speed up a reaction by lowering
the activation energy- the energy needed
for the reaction to start. (Although
reactions do happen at lower temperatures
they will be at a slower rate.)
4. The molecule that an enzyme acts on is
called its substrate. Each enzyme has a small area on
its surface called the active site. The new molecule
formed after the chemical reaction is called the product.
5. Enzymes are very specific because of the active site
which is different in each enzyme; therefore, cells
contain hundreds of different enzymes, each catalysing
a specific reaction.
6. Even a small concentration of enzyme can be very
effective as they remain unchanged after a reaction.

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7. Enzymes are extremely efficient: catalyse thousands of reactions in seconds.

8. Very high temperatures and extreme pH denature enzymes.
9. Enzymes are named according to the substrate on which they act. Proteases act
on proteins, carbohydrases on carbohydrates and lipases on fats (lipids).
10. The names of most enzymes end with –ASE. Ex: protease breaks down protein
molecules and lipase break down lipids. The table below shows some enzymes,
the substrates they work on and the products formed after the reaction.

✓ Metabolic reactions are of two types: anabolic and catabolic

✓ Some enzymes join two small molecules by a chemical bond and form a large
single molecule. Such reactions are called anabolic reactions.
Example: photosynthesis is an anabolic reaction as glucose is produced using
water and carbon dioxide.
✓ Some enzymes break a large single molecule chemically and form two or more
small molecules. Such reactions are called catabolic reaction.
Example: respiration is a catabolic reaction as glucose is broken down to
form water and carbon dioxide.

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Intra-cellular and extra-cellular enzymes

• Enzymes can be divided into two groups: intracellular and extracellular.

• Enzymes that are made and retained in the cell are known as
intracellular enzymes. They are found in the cytoplasm,
organelles or the nucleus. Example: Enzymes involved in
respiration.
• Enzymes that are produced in the cell, then packed and
secreted out of the cell are called extracellular enzymes.
• Extracellular enzymes catalyse their reactions outside the
cell. Most digestive enzymes are extracellular enzymes.

Example, amylase, lipase, etc.

1. Fungi and bacteria release extracellular enzymes in order to

digest their food.
2. A mould growing on a piece of bread releases starch-digesting
enzymes onto the bread and absorbs the soluble sugars that
the enzyme produces.
3. In the digestive systems of animals, extracellular enzymes
are released into the stomach and intestines to digest the food.

MECHANISM OF ENZYME ACTION:

❖ The ‘lock and key’ model is used to describe how enzymes work.
❖ The active site of each enzyme is very
different from the other. This is the reason
enzymes are said to be specific.
❖ The substrate fits exactly into the active site.
Therefore, we say the enzyme is
complementary to its substrate.
❖ The substrate fits into the active site of the
enzyme rather like a key fitting into a lock.

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❖ When the substrate attaches to the active site of

the enzyme, an enzyme substrate-complex is
formed.
❖ When the substrate joins up with the active site,
it lowers the energy needed for the reaction to
start, allowing the products to be formed more
easily.
❖ After the reaction, the enzyme is free to act on more substrate molecules.

In this example, the

substrate-sucrose is the key
& the enzyme-sucrase is the
lock. Try to use lock and key
model to describe the
mechanism of the given
reaction.

IMPORTANCE OF ENZYMES

1. Enzymes are important in

metabolic reactions, which sustain life.
2. Some examples of such reactions include photosynthesis (synthesis of glucose in
plants), respiration (break down of glucose to release energy), detoxification
(breakdown of toxic substances), synthesis of proteins, digestion of food etc.
3. Enzymes are also useful in industry, including food industry such as baking,
brewing, cheese making, etc.
4. Another example is the enzyme lactase. It breaks down the sugar lactose, found in
milk, into two other sugars called glucose and galactose. This is done because many
people are not able to digest lactose, so lactose can make them ill.
5. Enzymes are needed in seed germination. For example, bean seeds contain a lot
of starch, an insoluble carbohydrate. When the seed begins to germinate, the
enzyme amylase is secreted. This breaks the starch into maltose, which is soluble.

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Maltose is absorbed by the growing embryo and is used to get energy for
germination.
6. Enzymes are added in washing powders to remove stains like blood, egg yolk, etc.
which contain proteins.

A student investigated the effect of biological washing powder on stained clothes. Four
pieces of dry cloths that had been stained with the same food were treated as follows.

1. Washed using detergent with biological washing powder (labelled bio)

2. Washed using detergent without biological washing powder (labelled non-bio)
3. Washed using only water (labelled water)
4. No treatment (labelled not washed)

The figures below show the four cloth pieces

before and after the treatment. The cloth
piece washed with biological washing powder
became clear without any stain.

7. Enzymes are very important in breaking down toxic substances. For example,
Hydrogen peroxide is a toxic waste product produced from the metabolic reactions in
cells. Catalase enzyme in the liver breaks it down into harmless products, water
and oxygen. Catalase enzyme is also present in plant cells.
Liver cells have very high concentration of catalase enzyme.

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To investigate the activity of catalase small squares of filter paper were soaked in
fruit extracts and dried. The extract contains catalase enzyme. The filter papers
containing catalase enzyme were placed in hydrogen peroxide solution (H2O2) as shown
in the figure below.

✓ The catalase in the extract breaks down

H2O2 to produce water and oxygen.
✓ The pieces of filter paper rise to the
surface because of the oxygen bubbles
surrounding the filter paper which make
them light in weight.

Another investigation was carried out by adding seed

extract and hydrogen peroxide in a boiling tube and then
measuring the height of froth formed.

✓ The Froth is formed due to oxygen and water

production after the reaction.
✓ Crushing the seeds release the contents inside and
increases the surface area to volume ratio, thus
increasing the rate of the reaction.

Another investigation was carried out to find the effect of

cooking on the activity of catalase in potato tissue using
hydrogen peroxide. Two equal sized potatoes were taken
and one was boiled to cook and the second was kept as it
is. Each one was added to hydrogen peroxide.
The result is shown in the figure on the right.
✓ A Large amount of froth is produced in the uncooked
potato as the enzymes were active & broke down
hydrogen peroxide releasing oxygen and water.

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8. Enzymes are used in the extraction of juice from fruits. For example, an
enzyme called pectinase is used to produce clear fruit juice. This process is called
clarification.

• The figure on the right shows the volume of

juice extracted from two equally sized samples
of crushed apple over 16 minutes.
• Sample A contained pectinase enzyme and
Sample B contained the same volume of
water.
• In Sample A more juice is extracted and the
juice is clear. The process of extraction gives
more profit in less time.

A student investigated the effect of different pectinase concentrations on the

clarification of apple juice. The concentration of pectinase was the highest in P1 and
the lowest in P4. Figure 1 show the test tubes at the beginning and figure 2 shows the
final result.

Figure 1 Figure 2
Based on the results shown we conclude that the higher the concentration of
pectinase enzyme, the higher the clarity of the juice and vice versa.

FACTORS AFFECTING ENZYMES

The rate of enzyme activity is affected by the following factors:

1. Enzyme concentration
2. Substrate concentration
3. Temperature
4. pH

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1. EFFECT OF ENZYME CONCENTRATION ON ENZYME ACTIVITY.

The rate of an enzyme-catalyzed reaction is directly dependent on the enzyme

concentration. As the enzyme concentration increases, there are more free
active sites for substrate molecules to bind with, so the rate of the reaction
increases.

2. EFFECT OF SUBSTRATE CONCENTRATION ON ENZYME ACTIVITY.

As substrate concentration increases the rate of enzyme activity increases. This

is because there are more substrate molecules available for free enzyme active sites
to bind with. However, after some time when all the enzyme active sites are
occupied by substrates, the rate of the reaction remains constant, so increasing
the concentration of substrate doesn’t affect the rate of the reaction.

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3. EFFECT OF TEMPERATURE ON ENZYME ACTIVITY.

• At low temperatures, the enzyme molecules and the substrate molecules move very
slowly, so the rate of the reaction will be slow as the enzymes become inactive.
• At higher temperatures, the kinetic energy increases, so they move faster and
collide more frequently with each other. Therefore, rate of reaction increases due to
more effective collisions with an increase in temperature.
• The temperature at which an enzyme works best is called its optimum temperature.
However, when temperature rises above the optimum, the rate of reaction begins to
slow down.
• This is because higher temperature disrupts and
breaks the hydrogen bonds which hold the
enzyme molecule together giving it a specific
shape.
Therefore, the enzyme molecules and the active
site lose shape, so the substrate no longer fits
into the active site. The heat destroys the enzyme.
We say that the enzyme is denatured.
• By a temperature of 60 °C, the enzyme molecules
are completely denatured, and the active site is
no longer complementary to its substrate, therefore the reaction will stop.

✓ Most of the enzymes that work inside our bodies have an optimum temperature of around
37°C.
✓ Enzymes from plants often have an optimum temperature around 25 °C.
✓ Enzymes from bacteria that live in hot springs can have very high optimum temperatures,
often above 70 °C.
✓ Enzymes are not denatured at low temperatures- they become inactive and will work again
when the temperature is suitable.
✓ Once an enzyme is denatured, the damage is permanent.

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• OVERALL TREND
• At extremely cold temperatures, enzymes are inactivated/work slowly.
• Warm temperatures generally increase the relative rate (speed) of an enzyme.
• At extremely high temperatures, enzymes get denatured.
• The effect of temperature on the action of an enzymes is seen on a graph, where we
plot the rate of the reaction controlled by an enzyme against the temperature.

The figure on the left shows a denatured enzyme. Note that, the enzyme’s active site
is no longer complementary and cannot bind to the substrate due to excessive heat.

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EXPERIEMENT: Investigating effect of temperature on enzyme activity

1. Amylase is an enzyme that digests starch into maltose
2. The presence of starch can be tested by using iodine
solution. If there is starch in a solution, iodine turns
the solution into blue-black.
3. Starch solution is heated to a set temperature.
4. Iodine is added to the wells of a spotting tile.
5. Amylase is added to the starch solution and mixed
well.
6. Immediately the timer is started.
7. Every minute, droplets of solution are added to a new well of iodine solution.
8. This is continued until the iodine stops turning blue-black (this means there is
no more starch left in the solution as the amylase has broken it all down).
9. The experiment is repeated at different temperatures and the time taken for the
reaction to be completed is recorded at each temperature.
10. The less time the iodine solution takes to remain yellow-brown, the quicker the

starch was digested and so the better the enzyme works at that temperature.

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4. EFFECT OF PH ON ENZYME ACTIVITY.

• The pH of a solution is a measure of how acidic or alkaline it is. The shape and
activity of an enzyme is affected by pH.
• The pH inside cells is near neutral (pH 7) and most enzymes work best at this pH.
• Above or below this pH, the enzyme activity decreases, as shown in the graph.
• When the pH becomes more acidic or more alkaline, (at extremes of pH either side of
neutral,) the ionic bonds get disrupted and the enzyme loses its shape. Therefore,
the enzyme gets denatured and the active site is no longer complimentary to its
substrate.

• For most enzymes, there is a small pH range in which they can catalyse the reaction.
TREND
• The optimal pH for most of the enzymes in our body is ~pH 7.
• However, there are exceptions. The graph and the table below show the pH of
different parts of the alimentary canal.

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EXPERIEMENT: Investigating effect of pH on enzyme activity

1. Place drops of iodine solution in rows on a spotting tile.
2. Use the syringe to place 2cm3 of amylase into the test tube. Add 1cm3 of a buffer
solution (pH7) to the test tube using a syringe.
Use another test tube to add 2cm3 of starch solution to the amylase and buffer
solution and mix using a pipette.
3. Immediately start the timer.
4. After 10 seconds, use a pipette to place one drop of the mixture on the first drop
of iodine, which will turn to blue-black. Wait another 10 seconds and place
another drop of the mixture on the second drop of iodine.
5. Repeat every 10 seconds until the iodine solution remains yellow-brown.
6. Repeat the experiment at different pH values – the less time the iodine solution
takes to remain yellow-brown, the quicker the starch was digested and so the
better the enzyme works at that pH.

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EXPERIEMENT: Investigating effect of pH on protease enzyme in gelatin

• To measure the effect of pH on enzyme activity, protease enzyme was mixed with
solutions of different pH values. The mixtures were placed in holes cut in the
centre of dishes that contain gelatin.
• Gelatin is a protein which can be stained with a coloured dye. When the protease
digests the gelatin a colourless zone will form.
• Fig.1 shows a dish at the start of the experiment. Fig.2 shows the dishes after a
period of 24 hours.

❖ How could you improve the experiment to get more reliable results?
✓ Use a larger range of pH.
✓ Repeat the experiment more than once.
✓ Use a colorimeter to judge the colour change.
Pepsin is a protease enzyme found in the stomach where the pH is acidic. In this
experiment, 4 test-tubes where filled each with 1 cm3 cube of cooked egg white and
pepsin. They were placed at different pH. The time taken for the complete digestion
of the cubes of cooked egg white was measured.
Question: In which test tube will digestion occur faster?

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EXPERIEMENT: Investigating browning of apples

1. Obtain an apple cut in half from top to bottom.
2. Take one apple half and place it’s cut side down into a bowl of lemon juice. Leave
it for 2 minutes. The other apple half is exposed to air without adding lemon
juice. This is the control.
3. Leave the apples for about 30 minutes. Note any colour changes.

What Happened:

• When an apple is cut open, an enzyme called polyphenol oxidase is released

from the cells of the apple and reacts with the oxygen in the air. This
reaction causes the fruit to turn brown.
• Lemon juice prevents the apple from browning. Polyphenol oxidase works best
when the pH level is between 5.0 and 7.0. The pH of lemon juice is around 2, so
the enzyme become denatured, preventing the reaction from taking place.

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TYPES OF VARIABLES:
Scientific investigations include the following variables:

1. Independent variable
The variable that is changed in the experiment is called
the independent variable. This is the variable that is
manipulated/tested in an experiment.
Every experiment will have one independent variable.
For example, in an experiment to test the effect of
fertiliser on plant growth, amount of fertiliser is the
independent variable.
This variable comes on the X-axis in the graph.
2. Dependent variable
The variable that is measured in the experiment is
called the dependent variable. This is the variable that we
measure/observe/appears as a result.
For example, in the above experiment, the height of the
plant is the dependent variable.
This variable comes on the Y-axis in the graph.
3. Controlled variable
The variable that is kept constant in the experiment is called the controlled
variable.
Every experiment will have more than one constant variable.
For example, in the above experiment, type/species of plant, type of soil,
temperature, amount of water and sunlight, etc. are all controlled variables.
4. Control
Control is a setup where the independent variable is absent/eliminated.
For example, in the above experiment, a setup without any fertiliser would be a
control. However, all the variables are the same as in the main setup.
Importance of control:
When conducting experiments, having a control allows the results to be compared
with the main experiment. Hence, the researcher can validate if it was the variable
tested (independent variable) that was responsible for the outcome or not.

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Important terms and their definitions

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PAST PAPERS QUESTIONS LIST

Year Paper-1 Paper-2 Paper-3 Paper-6

Jun-01 9,10
Nov-01 10,11 4b
Jun-02 9,10
Nov-02 10,11
Jun-03 10,11
Nov-03 9,10 6
Jun-04 10,11 2
Nov-04 11,12 1a-c
Jun-05 12,13,16 6a,b,c
Nov-05 12,13
Jun-06 11,12
Nov-06 10,11
Jun-07 11,12 7
Nov-07 11,12 1a,b
Jun-08 12 3/31
Nov-08 12 3c/31
Jun-09 11 3a-c/31
Nov-09 11 3
Jun-10 11,14
Nov-10 10,11 1b/22 3/33 1/61
Jun-11 12,13 9/22 1/61
Nov-11 11,12 4/31 1/61
Jun-12 11,12/12 8/21 3c-e/31 1/62
Nov-12 11,12/11;12,13/ 4a,b/32 1a-d/62;3/63
12
Jun-13 16/11;11-13/12 5/23 5/31 1/63

Nov-13 12-14/11 9/21 1/62;1/63

Jun-14 11/11;11,12,12 5/23 2d,e/31 1/61;1/63

Nov-14 10,11/11 6a-c/31 1/62

Jun-15 12,13/11;11,12/ 8/21;2/22;3/23 1/61

12
Nov-15 12/11;11/12 1/61;1/63

CORE
Year Paper 1 Paper 3
June 16 10,11/11;11/12;10,11/13 7/33;
Nov 16 8/12;8/13 5b/33
March 17 9/12 1/32
June 17 7/11;7/12;7/13 1b/32;4b,5a/33
March 18 9/12 6/32

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June 18 10/11,12,13 8/31

Nov 18 10/11,12,13 8/31,5c/32,6/33
March 19 10,12 -
June 19 10/11,12,13 1c/63
EXTENDED
Year Paper 2 Paper 4 Paper 6
June16 10,11/21;9,10/22;9,10/23 1a-e/61
Nov 16 8/22;8/23 3/41;1b/42;2/43 1/63
March 17 9/22
June 17 7/21;7/22;7/23 2d/43
March 18 9/22 6/42
June 18 10/21,22,23 1a,b/41 1/61
Nov 18 9-10/21,22,23 6b/41 1/62;1/63
March 19 10/21,22 - 1/62
June 19 10/21,22,23 - -

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Choose the correct answer

1

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10

11

12

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Answer the following questions

1

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(b)

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7.

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