Enzymes are important proteins that control chemical reactions in all living organisms.

Each enzyme is specific to an individual reaction and requires certain conditions to

function efficiently.

What Are Enzymes?

• Definition: Enzymes are biological catalysts that speed up chemical reactions in

living organisms without being changed or used up.

• Composition: Enzymes are made up of long chains of amino acids that are folded
into specific three-dimensional shapes.

• Active Site: Each enzyme has an active site where the substrate (the substance
involved in the reaction) binds. The shape of the active site is complementary to
the substrate, ensuring specificity (lock-and-key model).

All living organisms perform a range of different chemical reactions all the time which
need to be tightly controlled. The sum of all these reactions in an organism or cell is called
metabolism. The rate of a reaction can usually be controlled by changing the
temperature but cells of the body will get damaged if the temperature is raised too high.
Therefore, living organisms use large proteins called enzymes to speed up reactions.

Enzymes are known as biological catalysts which means they increase the rate of
biological reactions without being changed or used up.

Enzymes are made up of long chains of amino acids that are folded into specific shapes.

All enzymes have an active site which is where it can bind to a substrate (substance
involved in the reaction).

Enzymes and their active sites have very specific shapes. The specific shape of each
active site means only certain substrates can bind to it and form enzyme-substrate
complexes. This means each enzyme can only catalyse one specific type of reaction
and therefore produce the products.

The ‘lock and key’ model is a simplified version of how enzymes work. It states that the
active site of an enzyme fits the substrate perfectly like a lock and a key, they are
complementary.

Enzymes as Biological Catalysts

• Enzymes are proteins that act as biological catalysts to speed up the rate of a
chemical reaction without being changed or used up in the reaction

• They are biological because they are made in living cells

• Enzymes are necessary to all living organisms as they maintain reaction speeds of
all metabolic reactions at a rate that can sustain life
 o For example, if we did not produce digestive enzymes, it would take around
2 - 3 weeks to digest one meal; with enzymes, it takes around 4 hours

o Often the products of one reaction are the reactants for another (and so
on)

The mechanism of enzyme action

Figure 1: Enzyme substrate specificity

• Enzymes are specific to one particular substrate (molecule/s that get broken
down or joined together in the reaction) as the enzyme is a complementary
shape to the substrate

• The product is made from the substrate(s) and is released

 Figure 2: Enzyme specificity: lock and key model of enzyme activity

• Enzymes are specific to one particular substrate(s) as the active site of the
enzyme, where the substrate attaches, is a complementary shape to the substrate

• When the substrate moves into the enzyme’s active site they become known as
the enzyme-substrate complex

• After the reaction has occurred, the products leave the enzyme’s active site as
they no longer fit it and it is free to take up another substrate

o Step One: Enzymes and substrates randomly move about in solution

o Step Two: When an enzyme and its complementary substrate randomly

collide an enzyme-substrate complex forms, and the reaction occurs

o Step Three: A product (or products) forms from the substrate(s) which are
then released from the active site. The enzyme is unchanged and will go on
to catalyse further reactions
Importance of Enzymes

1. Metabolism: Enzymes control the chemical reactions in metabolism, the sum of

all reactions in a cell or organism.

2. Efficiency: They allow reactions to occur at body temperature, avoiding damage

caused by high temperatures.

3. Reusability: Enzymes are not consumed in reactions, so they can be reused

multiple times.

Examples of Enzymes and Their Roles

1. Amylase: Breaks down starch into maltose.

o Secreted by: Salivary glands and pancreas.

2. Protease: Breaks down proteins into amino acids.

o Secreted by: Pancreas.

3. Lipase: Breaks down fats into fatty acids and glycerol.

o Secreted by: Pancreas.

Enzyme-Substrate Interaction

The process can be described in three steps:

• The substrate binds to the enzyme’s active site.

• The enzyme catalyzes the reaction, converting the substrate into products.
• The products are released, and the enzyme is free to catalyze another
reaction.

Enzyme Substrate Product(s) Site of Secretion

Amylase Starch Maltose Salivary glands, pancreas
Protease Proteins Amino acids Pancreas
Lipase Fats (lipids) Fatty acids + Glycerol Pancreas
Denaturing of Enzymes

Enzymes are proteins and have a specific shape, held in place by bonds.

This is extremely important around the active site area as the specific shape is what
ensures the substrate will fit into the active site and enable the reaction to proceed.

The rates of enzyme-controlled reactions are affected by temperature and pH.

Enzymes work fastest at their ‘optimum temperature’ – in the human body, the optimum
temperature is 37⁰C.

Heating to high temperatures (beyond the optimum) will break the bonds that hold the
enzyme together and it will lose its shape -this is known as denaturation.

Substrates cannot fit into denatured enzymes as the shape of their active site has been
lost.

Denaturation is irreversible - once enzymes are denatured they cannot regain their
proper shape and activity will stop.
Factors Affecting Enzyme Activity

Temperature

Increasing the temperature from 0⁰C to the optimum increases the activity of enzymes as
the more energy the molecules have the faster they move and the number of
collisions with the substrate molecules increases, leading to a faster rate of reaction.

This means that low temperatures do not denature enzymes, they just make them work
more slowly.

Increasing the temperature will increase

the rate of reaction because the enzymes
and substrate will have more kinetic energy
and so will be more likely to collide and
react.

This will only happen up to a point (the

optimum temperature). After this, bonds in
the enzyme will begin to break, they will lose
their specific shape and become
denatured.

Figure 3: Graph showing the effect of temperature on the rate of enzyme activity
When the shape of the active site changes, substrates will not be able to bind, enzyme-
substrate complexes won’t be able to form and the enzyme will stop catalysing the
reaction.

• Enzymes have a specific optimum temperature – the temperature at which they

catalyse a reaction at the maximum rate

• Lower temperatures either prevent reactions from proceeding or slow them

down this is because:

o Molecules move relatively slowly at lower temperatures

o Therefore there is a lower frequency of successful collisions that occur

between substrate molecules and the active site of the enzyme

o So there are less frequent enzyme-substrate complexes formed

o Substrates and enzymes collide with less energy, making it less likely for
bonds to be formed or broken (stopping the reaction from occurring)

• Higher temperatures speed up reactions this is because:

o Molecules move more quickly at higher temperatures

o Which results in higher frequency of successful collisions between

substrate molecules and the active sites of enzymes

o So there are more frequent enzyme-substrate complexes formed

o Substrates and enzymes collide with more energy, making it more likely
for bonds to be formed or broken (allowing the reaction to occur)

• However, as temperatures continue to increase, the rate at which an enzyme

catalyses a reaction drops sharply, as the enzyme begins to denature:

o Bonds (e.g. hydrogen bonds) holding the enzyme molecule in its precise
shape start to break

o This causes the tertiary structure of the protein (i.e. the enzyme)
to change

o This permanently damages the active site, preventing

the substrate from binding

o Denaturation has occurred if the substrate can no longer bind

o Very few human enzymes can function at temperatures above 50°C

 ▪ This is because humans maintain a body temperature of about
37°C, therefore even temperatures exceeding 40°C will cause the
denaturation of enzymes

Figure 4: The effect of temperature on the rate of an enzyme-catalysed reaction. Enzyme activity will have an optimum
temperature specific for each enzyme.

pH:

The optimum pH for most enzymes is 7 but some that are produced in acidic conditions,
such as the stomach, have a lower optimum pH (pH 2) and some that are produced in
alkaline conditions, such as the duodenum, have a higher optimum pH (pH 8 or 9).

If the pH is too high or too low, the bonds that hold the amino acid chain together to make
up the protein can be destroyed

This will change the shape of the active site, so the substrate can no longer fit into it,
reducing the rate of activity

Moving too far away from the optimum pH will cause the enzyme to denature and activity
will stop
Enzymes also have an optimum
pH. If the pH is too high or too low
then the bonds in the enzyme will
start to break, they will lose their
specific shapes and they will
become denatured.

If the active site changes shape,

substrates will not be able to bind,
enzyme-substrate complexes
won’t form and so the enzyme will
stop catalysing the reaction.

Each type of enzyme will have a different optimum pH. Pepsin is an enzyme that breaks
down proteins and works best in the acidic conditions of the stomach.

• All enzymes have an optimum pH which is a pH at which they operate best

• Enzymes are denatured at extremes of pH

o Hydrogen and ionic bonds hold the tertiary structure of the protein (i.e.
the enzyme) together

o Below and above the optimum pH of an enzyme, solutions with an excess

of H+ ions (acidic solutions) and OH- ions (alkaline solutions) can cause
these bonds to break

o This alters the shape of the active site, which means enzyme-substrate
complexes form less easily
 o Eventually, enzyme-substrate complexes can no longer form at all

o At this point, complete denaturation of the enzyme has occurred

• Where an enzyme functions, can be an indicator of its optimal environment:

o E.g. pepsin is found in the stomach, an acidic environment at pH 2 (due to

the presence of hydrochloric acid in the stomach’s gastric juice)

o Pepsin’s optimum pH, not surprisingly, is pH 2

Figure 5: The effect of pH on the rate of an enzyme-catalysed reaction for three different enzymes (each with a different
optimum pH)

• When investigating the effect of pH on the rate of an enzyme-catalysed reaction,

you can use buffer solutions to measure the rate of reaction at different pH
values:

o Buffer solutions each have a specific pH

o Buffer solutions maintain this specific pH, even if the reaction taking place
would otherwise cause the pH of the reaction mixture to change

o A measured volume of the buffer solution is added to the reaction mixture

 o This same volume (of each buffer solution being used) should be added
for each pH value that is being investigated

Temperature can both affect the speed at which molecules are moving (and therefore the
number of collisions between enzyme and substrate in a given time) and can denature
enzymes (at high temperatures).

pH, however, does not affect collision rate but only disrupts the ability of the substrate to
bind with the enzyme, reducing the number of successful collisions until eventually, the
active site changes shape so much that no more successful collisions can occur.

Substrate Concentration:

• The greater the substrate concentration, the higher the rate of reaction:

o As the number of substrate molecules increase, the likelihood of enzyme-

substrate complex formation increases

o If the enzyme concentration remains fixed but the amount of substrate is

increased, past a certain point all available active sites eventually
become saturated and any further increase in substrate concentration
will not increase the reaction rate

o When the active sites of the enzymes are all full, any substrate molecules
that are added have nowhere to bind in order to form an enzyme-
substrate complex and so the reaction rate will not increase any further
until active sites become free again

• For this reason, in the graph below there is a linear increase in reaction rate as
substrate is added, which then plateaus when all active sites become occupied
 Figure 6: The effect of substrate concentration on the rate of an enzyme-catalysed reaction

If substrate concentration is continually increased but enzyme concentration is kept

constant, there eventually comes a point where every enzyme active site is working
continuously. At this point, the substrate molecules are effectively ‘queuing up’ for an
active site to become available. At this stage, the enzyme is working at its maximum
possible rate, known as Vmax (V stands for velocity).

Enzyme Concentration:

o More enzymes can increase the reaction rate if substrate is available.

o Enzyme concentration affects the rate of reaction

o The higher the enzyme concentration in a reaction mixture, the greater the
number of active sites available and the greater the likelihood of enzyme-
substrate complex formation

o As long as there is sufficient substrate available, the initial rate of

reaction increases linearly with enzyme concentration

o If the amount of substrate is limited, at a certain point any further increase

in enzyme concentration will not increase the reaction rate as the amount
of substrate becomes a limiting factor
 Figure 7: The effect of enzyme concentration on the rate of an enzyme-catalysed reaction

Investigating the effect of pH on the rate of reaction.

The reaction used for this experiment is the breakdown of starch into maltose by the
enzyme amylase. Starch can be easily detected because if added to iodine solution it
will turn a blue-black colour. If starch is not present, the iodine solution will remain a
brown-orange colour. This makes it very useful for investigating enzymatic reactions.

Doing the experiment

1. Using a pipette, prepare spotting tiles by placing a drop of iodine solution in each
cavity.
2. Prepare a test tube containing 2 𝑐𝑚3 of amylase(enzyme) and 1 𝑐𝑚3of a buffer
solution with a known pH. Place this in a water bath at 35° C .
3. Prepare another test tube with 2 𝑐𝑚3 of starch solution and place it in the water
bath.
4. Pour the starch into the amylase and buffer solution and start the timer. Make
sure to keep the test tubes in the water bath.
5. Every 10 second take one drop of the solution and place it on the iodine on the
spotting tile. The colour will turn blue-black when starch is present.
 6. Continue until the colour no longer turns blue-black and remains an orange-
brown colour. At this point all the starch has been digested to maltose. The
quicker it takes for the spot to remain orange-brown, the faster the rate of reaction.
7. Repeat the experiment using buffers of different pH values to find the effect it has
on the rate of breakdown of the starch.

It is important to keep the temperature of all the solutions the same, using the water
bath, as temperature affects the rate of reaction.

It may also be wise to carry out a control experiment to test if it is actually the amylase
that breaks down the starch. This could be done by replacing the amylase with distilled
water.

Calculating the rate of reaction

The time it takes for the starch to

breakdown and the colour to
remain brown-orange is not a
measure of the rate of reaction.

Rate is how much something

changes over a given time. As the
measure of starch breakdown in
this experiment is qualitative
(changes colour),not
quantitative (no numerical
values), we can use a constant
value for the measure of change.

For this experiment we use : 1000

1000
Rate of reaction (𝑠 −1 ) = 𝑇𝑖𝑚𝑒 ( 𝑠 )

Now you can plot a graph to show how pH affects the breakdown of starch by amylase.
Applications of Enzymes

a) Digestion:

a. Enzymes break down food into smaller molecules for absorption (e.g.,
carbohydrates into glucose).

b) Industry:

a. Enzymes like proteases are used in detergents to remove protein stains.

c) Medicine:

o Enzymes are used in diagnosing diseases (e.g., blood glucose monitoring).

Use of enzymes in industries

Biological detergents

Biological detergents contain protein-digesting enzymes produced by genetically

engineered bacteria. Many of the stains on clothes, like blood and sweat, are proteins.
Biological detergents have a number of advantages:

• The enzymes work at relatively low temperatures.

• They remove stains which would otherwise need high temperature washes.

• Energy and money are saved by allowing low temperature washes.

• They help to clean delicate fabrics which would otherwise be damaged by a hot
wash.

Cheese making

A mixture of enzymes called rennet is used during the production of cheese. Rennet is
added to milk and helps to separate it into solid and liquid parts. Rennet can come from
animal, fungal or plant sources, or can be produced by genetically modified bacteria.

Enzyme immobilisation and continuous-flow processing

In batch processing all the raw materials and the enzyme or cells are put into a vessel
called a fermenter at the start. At the end the product and the enzyme must to be
separated.

Immobilisation techniques restrict the movement of enzymes. This is usually done by

attaching the enzymes to a solid substance such as a bead.

Immobilisation techniques allow continuous flow processing by holding the enzymes or

cells in a column in a reactor vessel. The raw materials are fed into the top of the column
continuously and the product is continuously removed from the bottom. This results in
increased productivity and reduced costs compared to batch processing, as the process
can be run non-stop for a long period of time.

Enzymes are used in biological washing powders.

(i) How enzymes increase the efficiency of biological washing powders

Biological washing powders contain enzymes such as proteases, lipases, and

amylases, which help break down stains. These enzymes speed up the breakdown of
large, insoluble molecules into smaller, soluble ones, making stains easier to remove.

• Proteases break down protein-based stains (e.g., blood, sweat, egg).

• Lipases break down fat and oil stains (e.g., grease, butter).

• Amylases break down starch-based stains (e.g., food, sauces).

Because enzymes work at lower temperatures, biological washing powders allow for
effective cleaning with less energy use, making them more efficient and eco-friendly.

(ii) Why the temperature of the wash needs to be carefully controlled

Enzymes are temperature-sensitive proteins. If the temperature is too low, the enzyme
activity is slow, meaning stains take longer to break down. However, if the temperature is
too high, enzymes denature, losing their shape and becoming ineffective. This means
the washing powder will not work properly.

(iii) Suitable temperature for a biological wash and explanation

A suitable temperature for a biological washing powder is 30–40°C.

• This is warm enough for enzymes to work effectively and speed up stain
breakdown.

• It is not too high, so enzymes do not denature, ensuring they remain active.

• Lower temperatures (e.g., 30°C) also save energy, making washing more
environmentally friendly.

( iv) How Enzymes Are Manufactured for Use in Biological Washing Powders

1. Selection of Microorganisms

o Enzymes for biological washing powders are produced using

microorganisms such as bacteria (e.g., Bacillus species) and fungi (e.g.,
Aspergillus species).
 o These microorganisms are chosen because they naturally produce
enzymes like proteases, lipases, and amylases, which help break down
stains.

2. Fermentation Process

o The selected microorganisms are grown in large fermenters under

controlled conditions.

o They are provided with nutrients (e.g., glucose, nitrogen, and minerals),
optimal temperature, oxygen levels, and pH to maximize enzyme
production.

o The microorganisms produce and secrete enzymes into the surrounding

liquid as they grow.

3. Extraction and Purification

o The enzyme-rich liquid is filtered to remove unwanted microbial cells.

o The enzymes are then purified through various techniques, such as

precipitation or ultrafiltration, to increase their effectiveness.

4. Stabilisation and Formulation

o The purified enzymes are mixed with stabilising agents (e.g., salts and
surfactants) to maintain their activity during storage.

o They are then dried into a powder or encapsulated to protect them from
damage before being added to the washing powder.

5. Testing and Quality Control

o The final enzyme product is tested to ensure it works effectively in washing

powders.

o Manufacturers check for enzyme activity, stability, and compatibility

with other ingredients in the detergent.

6. Packaging and Distribution

o The enzyme-based washing powder is packaged and distributed for

consumer use.

This method ensures the large-scale production of cost-effective, efficient, and

environmentally friendly biological washing powders.
Enzymes are biological catalysts. Fig. 3.1 shows how the enzyme, sucrase, breaks
down a molecule of sucrose.

Sucrase catalyses the breakdown of sucrose using the lock and key hypothesis:

1. Substrate binding – The sucrose molecule fits into the active site of the sucrase
enzyme, forming an enzyme-substrate complex (as shown in Fig. 3.1).

2. Hydrolysis reaction – Water (H₂O) is added, breaking the bond between glucose
and fructose within sucrose.

3. Product release – The glucose and fructose molecules are released from the
active site, and the enzyme remains unchanged, ready to catalyse another
reaction.

This process increases the speed of sucrose breakdown, making it available for use in
respiration.

Three enzymes, P, Q and R, were extracted from different regions of the alimentary
canal of a mammal. The effect of pH on the activity of the enzymes was investigated
at 40 °C. The results are shown in Fig. 3.2.
(i) Explain why the investigation was carried out at 40°C.

The investigation was carried out at 40°C because this is close to the optimum
temperature for enzymes in the human body. Most enzymes function best around body
temperature (37–40°C), allowing them to catalyse reactions at their maximum efficiency
without becoming denatured.

(ii) Using information in Fig. 3.2, describe the effects of increasing pH on the rate of
activity of enzyme Q.

• Enzyme Q shows maximum activity at pH 7, meaning it works best in neutral

conditions.

• As pH increases above 7, the enzyme activity gradually decreases, suggesting

the enzyme is becoming less efficient.

• Similarly, as pH decreases below 7, enzyme activity also declines, showing that

acidic conditions reduce its effectiveness.

• At very low or high pH values, enzyme Q is almost inactive, indicating that

extreme pH levels may denature the enzyme.

Some baby foods are manufactured by pre-digesting foodstuffs containing

carbohydrates, fats and proteins with enzymes. Describe the roles of different types
of enzymes in preparing these baby foods.
In the preparation of baby foods, enzymes are used to pre-digest carbohydrates, fats,
and proteins, making the food easier for infants to digest. The following types of enzymes
are used:

1. Carbohydrases (e.g., Amylase and Maltase)

o Break down carbohydrates into simpler sugars.

o Amylase converts starch into maltose, and maltase further breaks

maltose into glucose.

o This makes the food sweeter and easier to absorb.

2. Proteases (e.g., Pepsin and Trypsin)

o Break down proteins into smaller peptides and amino acids.

o This softens the food and makes proteins more accessible for the baby’s
digestion and growth.

3. Lipases

o Break down fats (lipids) into glycerol and fatty acids.

o This helps infants absorb essential fats and energy from food more easily.

By using these enzymes, baby food becomes softer, easier to digest, and more suitable
for an infant’s immature digestive system.

Enzymes extracted from bacteria are used in biological washing powders. Describe
how bacteria are used to produce enzymes for biological washing powders.

To produce enzymes for biological washing powders, bacteria are used in a controlled
industrial process. The steps involved are:

1. Selection of Bacteria – Scientists choose specific bacterial strains that naturally

produce enzymes like proteases (break down proteins), lipases (break down fats),
and amylases (break down starches).

2. Fermentation – The selected bacteria are grown in large fermentation tanks under
optimal conditions (warm temperature, suitable pH, and nutrient supply). They
multiply rapidly and produce enzymes as they grow.

3. Harvesting Enzymes – Once a high concentration of enzymes is produced, the

bacterial cells are removed by filtration or centrifugation, leaving behind the
enzyme-rich solution.

4. Purification – The extracted enzymes are purified to remove any unwanted

byproducts, ensuring they are safe and effective for use in washing powders.
 5. Drying and Formulation – The purified enzymes are dried into a powder or
granulated form, making them easy to mix with other ingredients in washing
powders.

6. Final Product – The enzyme powder is added to the biological washing powder,
allowing it to break down stains effectively at lower temperatures, making washing
more efficient and environmentally friendly.

This biotechnological process ensures a continuous and cost-effective supply of

enzymes for detergents.

Food and blood stains on clothes may contain proteins and fats. Explain how
enzymes in biological washing powders act to remove food and blood stains from
clothes.

Biological washing powders contain specific enzymes that help break down food and
blood stains on clothes. The main enzymes involved are:

1. Proteases – These enzymes break down protein molecules found in stains like
blood, egg, and meat. They work by hydrolyzing peptide bonds, converting large,
insoluble proteins into smaller, soluble amino acids, which dissolve easily in
water and are washed away.

2. Lipases – These enzymes break down fats and oils found in greasy food stains
(e.g., butter, cooking oil). They act by breaking ester bonds in fats, converting them
into glycerol and fatty acids, which are soluble in water.

How They Work:

• When clothes are washed in warm water (typically 30–40°C) with biological
detergent, the enzymes become active and attach to specific stain molecules.

• The enzymes catalyze hydrolysis reactions, breaking large, insoluble molecules

into smaller, soluble ones.

• These water-soluble products dissolve easily, allowing the stains to be rinsed

away.

Since enzymes function best at moderate temperatures, biological washing powders

allow effective stain removal at lower temperatures, saving energy and reducing fabric
damage.

Catalase is an enzyme found in plant and animal cells. It has the function of breaking
down hydrogen peroxide, a toxic waste product of metabolic processes. State the
term used to describe the removal of waste products of metabolism.

The term used to describe the removal of waste products of metabolism is excretion.
An investigation was carried out to study the effect of pH on catalase, using pieces
of potato as a source of the enzyme. Oxygen is formed when catalase breaks down
hydrogen peroxide, as shown in the equation.
𝑪𝒂𝒕𝒂𝒍𝒂𝒔𝒆
𝑯𝒚𝒅𝒓𝒐𝒈𝒆𝒏 𝑷𝒆𝒓𝒐𝒙𝒊𝒅𝒆 > 𝑾𝒂𝒕𝒆𝒓 + 𝑶𝒙𝒚𝒈𝒆𝒏e

The rate of reaction can be found by measuring how long it takes for 10 cm3 oxygen
to be collected.

State the independent (input) variable in this investigation.

The pH of the solution.

Suggest two factors that would need to be kept constant in this investigation.

Temperature – To ensure that enzyme activity is only affected by pH and not by

temperature changes.

Concentration or volume of hydrogen peroxide – To ensure that the amount of

substrate available for the reaction remains the same.

Starch, glucose and fructose are carbohydrates. Fructose syrup is used as a

sweetening agent as an alternative to sucrose. The flow chart in Fig. 3.1 shows how
fructose is prepared from maize starch.

i. Name enzyme
Amylase (or Carbohydrase) – This enzyme breaks down starch into glucose syrup.
 ii. State why it is necessary to adjust the pH before an enzyme is added to the
process.
Enzymes work best at their optimum pH. If the pH is too high or too low, the
enzyme may become denatured or less effective, slowing down the reaction.
Adjusting the pH ensures maximum enzyme activity for efficient conversion of
starch into glucose.
a) Why is it important that protease enzymes do not contaminate the glucose
syrup?
Protease enzymes break down proteins, which could damage or remove
important enzymes used in the process (e.g., amylase and isomerase). If
proteases contaminate the glucose syrup, they might break down the enzymes
needed to convert starch into glucose or glucose into fructose, reducing yield and
efficiency.
i. The formation of fructose syrup from glucose syrup is carried out at a
temperature of 60 °C. Suggest an important property of enzyme 2 that allows
it to be used at temperatures as high as 60 °C.

The enzyme must be heat-stable (or thermostable), meaning it does not denature or lose
its function at high temperatures. This allows it to remain active and efficient at 60°C,
ensuring the conversion of glucose into fructose continues effectively.

ii. Pectinase is an enzyme that breaks down compounds known as pectins. Cell
walls of fruits, such as apples and mangoes, contain pectins. Explain the
advantages of using pectinase in fruit juice production.

Increases juice yield – Pectinase breaks down pectins in the cell walls, helping to
release more juice from the fruit.

Produces clearer juice – Pectins cause cloudiness in juice, and breaking them down
makes the juice clearer and more visually appealing.

Speeds up the extraction process – The enzyme helps break down fruit tissues faster,
making juice production more efficient.

Improves filtration – Since pectin makes juice thick, breaking it down allows for easier
filtration and removal of unwanted solids.

Helicobacter pylori is a bacterium that infects the stomach and causes ulcers. The
bacteria secrete urease that helps them to colonise the stomach lining.

Explain why bacteria do not usually grow inside the stomach.

The stomach contains hydrochloric acid (HCl), which creates a highly acidic environment
(pH 1–2). This kills most bacteria by denaturing their proteins and disrupting their cell
membranes, preventing their growth and colonization.
However, Helicobacter pylori survives by producing urease, which helps neutralize the
stomach acid, allowing it to colonize the stomach lining.

This explanation includes key scientific terms and should be suitable for IGCSE Grade 9!
Let me know if you need any refinements.

How urease helps Helicobacter pylori colonize the stomach:

➢ Urease is an enzyme that breaks down urea (a compound found in the stomach)
into ammonia and carbon dioxide.

➢ Ammonia is alkaline, which neutralizes stomach acid around the bacteria,

creating a less acidic environment where H. pylori can survive and colonize the
stomach lining.

➢ This protection allows the bacteria to attach to the stomach lining and avoid
being destroyed by the acidic conditions.

How the immune system protects against infection by bacteria like H. pylori:

1. Physical barriers – The body’s first line of defense includes the mucus lining of
the stomach, which helps trap bacteria and prevent them from reaching stomach
cells.

2. White blood cells (WBCs):

o Phagocytes (e.g., macrophages & neutrophils) engulf and destroy

bacteria by phagocytosis.

o Lymphocytes produce antibodies that bind to bacterial antigens and help

neutralize or destroy them.

3. Inflammatory response – The immune system triggers inflammation, increasing

blood flow and bringing more immune cells to fight the infection.

4. Enzymes and chemicals – The body releases enzymes (like lysozymes) that
break down bacterial cell walls, helping to kill the bacteria.

Despite this immune response, H. pylori can evade the immune system by hiding under
the mucus lining and neutralizing stomach acid, making it difficult to eliminate without
antibiotics.

During stage 5 microorganisms break down organic matter consisting of cellulose,

starch, protein and lipid (fat). The microorganisms multiply during this stage and are
recycled. Complete Fig. 6.2 by writing in the boxes the names of the enzymes used
to catalyse the reactions shown. The first box has been completed for you.
Here are the enzymes that would complete the figure:

1. Starch → Maltose → Glucose: The enzyme is amylase.

2. Protein → Amino acids: The enzyme is protease.

3. Lipid (fat) → Fatty acids and glycerol: The enzyme is lipase.

State why it is important that sewage is treated.

It is important that sewage is treated to:

1. Prevent pollution: Untreated sewage can contaminate water bodies, causing

harm to aquatic life and the environment.

2. Prevent the spread of disease: Sewage contains harmful microorganisms that

can spread diseases such as cholera and typhoid if not treated.

3. Protect water sources: Proper treatment ensures that rivers, lakes, and
groundwater remain safe for drinking, recreation, and irrigation.

Explanation in terms of enzymes:

• Enzymes are crucial in the biological treatment of sewage because they speed up
the breakdown of complex organic substances like proteins, fats, and
carbohydrates into simpler, harmless products.

o Proteases break down proteins into amino acids.

o Lipases break down fats into fatty acids and glycerol.

o Amylases break down starch into sugars.

These simpler products are easier for microorganisms to digest, allowing for faster
decomposition and the removal of harmful substances from sewage. This makes the
treated water safe to return to the environment or reuse.

Microorganisms in the soil release enzymes to digest dead leaves.

(a) How enzymes catalyze chemical reactions:

Enzymes are biological catalysts that speed up chemical reactions by lowering the
activation energy required for the reaction to occur.

1. Enzymes have an active site that is specific to the substrate (the molecule they act
on).

2. The substrate binds to the enzyme's active site, forming an enzyme-substrate

complex.

3. The enzyme helps break down or build up the substrate into the products,
releasing them.

4. The enzyme remains unchanged after the reaction and can be reused.

By reducing the activation energy, enzymes allow reactions to occur faster and more
efficiently under normal environmental conditions, such as those found in soil.

Part of dead leaf cells digested by cellulase:

Cellulase digests cellulose, which is a major component of the cell walls of dead leaf
cells. The enzyme breaks down cellulose into simpler sugars like glucose, which
microorganisms can then use as a source of energy and nutrients.

Table 6.1 shows the results of a study comparing the decomposition of dead leaves at
two locations A and B.

i. Compare the enzyme activity at location A with the enzyme activity at

location B. You will gain credit for using the data from Table 6.1 to support your
answer.
Protease activity: The protease activity at location A (2750 µmol min⁻¹) is slightly higher
than at location B (2670 µmol min⁻¹), with a difference of 80 µmol min⁻¹.

Cellulase activity: The cellulase activity at location A (4790 µmol min⁻¹) is much higher
than at location B (2500 µmol min⁻¹), showing a significant difference of 2290 µmol min⁻¹.

Soil conditions:

• The pH at location A is 6.0, while at location B it is 3.5, which is more acidic.

• The soil water content is much higher at location B (77%) compared to location A
(10%).

ii. Possible reasons for differences in enzyme activity:

1. pH differences:

o The pH at location A (6.0) is closer to the optimal pH for most enzymes,

which typically function best in neutral to slightly acidic conditions.

o At location B, the lower pH (3.5) may denature the enzymes or reduce their
activity.

2. Soil water content:

o The higher water content at location B (77%) may lead to waterlogging,

reducing oxygen availability. This can slow down microbial activity and
enzyme production.

o Location A has a lower water content (10%), which may provide a more
favorable environment for enzyme activity.

3. Environmental conditions:

o The combined effect of extreme acidity and high water content at location
B creates less favorable conditions for enzyme function and microbial
activity, leading to lower enzyme activity overall.

Enzymes are necessary for many biological processes, such as the digestion of fat.
𝑳𝒊𝒑𝒂𝒔𝒆
𝑭𝒂𝒕 + 𝑾𝒂𝒕𝒆𝒓 > 𝑭𝒂𝒕𝒕𝒚 𝑨𝒄𝒊𝒅 + 𝑮𝒍𝒚𝒄𝒆𝒓𝒐𝒍

i. Explain why enzymes are necessary for biological processes.

Enzymes are biological catalysts that speed up chemical reactions by lowering the
activation energy required. Without enzymes, biological processes like digestion
would occur too slowly to sustain life. Enzymes ensure that the breakdown of
 complex molecules, such as fats, proteins, and carbohydrates, happens
efficiently and quickly, providing organisms with essential nutrients and energy.
ii. Lipase, protease and amylase are enzymes secreted into the alimentary
canal. Name one organ that secretes each enzyme. Choose your answers
from this list.
gall bladder colon pancreas liver salivary glands rectum
You can use each organ only once.
Organs that secrete lipase, protease, and amylase:
a) Lipase: Secreted by the pancreas.
b) Protease: Secreted by the pancreas.
c) Amylase: Secreted by the salivary glands.
(Note: While the pancreas secretes lipase and protease, the salivary glands
produce amylase for carbohydrate digestion in the mouth.)