CAPE BIOLOGY UNIT 1 ENZYMES TUTORIAL (PART 1)

1. What are enzymes? (3 mks)

Enzymes are globular protein that catalyze metabolic reactions.
They are also called biological catalyst- speed up a chemical reaction, remain unchanged at the end.
All enzymes are protein molecules made by living cells

2. State five properties of enzymes. (5 mks)

⚫ All are globular proteins coded for by DNA

⚫ Presence does not alter the reaction or the products formed
⚫ Very efficient (little enzyme needed to bring about a reaction)
⚫ Highly specific – only catalyze a single type of reaction

⚫ Catalyze reversible reactions

⚫ Affected by pH, temperature, enzyme and substrate conc.
⚫ Lower activation energy – energy required to start the chemical reaction.
⚫ Possess active sites.
1. All enzymes are globular proteins.
2. Enzymes only change the rate of a reaction.
3. Enzymes are present in very small amounts.
4. Enzymes are specific to the reaction that they catalyse.
5. Enzyme-catalyzed reactions are affected by the amount of substrate present.
6. The role of an enzyme catalyzed reaction is affected by temperature in a characteristic way.
7. pH has a major effect on enzyme activity.

3. What don’t enzymes affect? (2 mks)

They don't affect the final energy of the products; they do not change the equilibrium of the reaction.

Complete the following description of enzymes by adding the most appropriate word or words in
the gaps:
 CAPE BIOLOGY UNIT 1 ENZYMES TUTORIAL (PART 1)

Enzymes are biological catalysts which increase the rate of a reaction by reducing the
_________activation________energy needed for the reaction to take place. Enzymes are globular
_______proteins_________ which possess a region on their surface where binding to a substrate
occurs. This region is called the ____active site________. The bonds which maintain the tertiary
structure of the enzyme can be disrupted by factors such as _____ hydrophilic R group
_____________ and _______their side chains,
____________, reducing the catalytic action of the enzyme. Ions of heavy metals such as mercury
cause ___competitive _____________inhibition of enzymes. Other inhibitors reduce enzyme
activity because they are a similar shape to the enzyme’s substrate. These are called
_____________non competitive_____ inhibitors. (7 mks)

4. What is meant by enzyme ‘cofactor’? Explain how a vitamin deficiency may affect enzyme
activity. (3 mks)

A cofactor is anon-proteinchemicalcompound or metallic ion that is required for protein's

biological activity to happen.They assist in the biochemical reaction E.g FAD or Copper

Cofactor=non-protein necessary component (e.g iron ion). It is needed for the function of the
protein or enzyme. Coenzyme=complex cofactor (organic). It is needed for the function of the
enzyme; they bind loosely to the enzyme. Prosthetic groups are cofactors that bind tightly to
proteins or enzymes and are necessary for their function; they may be organic or metallic ions
(for e.g. haem and iron).

5. Describe the three main categories of chemical reactions. (3 mks)

Metabolism: Sum of all the chemical reactions in the cells

Catabolism: breakdown of (macro) molecules via hydrolysis, usually accompanied by release of

energy
A🡪 B + C

Anabolism: synthesis of (macro) molecules via condensation usually requiring energy

D+ E🡪 F

6. What is meant by activation energy? (1 mk)

The activation energy is the energy required to start a reaction. Enzymes are
proteins that bind to a molecule, or substrate, to modify it and lower the energy required
to make it react.
 CAPE BIOLOGY UNIT 1 ENZYMES TUTORIAL (PART 1)

7. Describe how enzymes work to lower the activation energy of a reaction? Be sure to include a
graphical representation. (6 mks)

Enzymes will lower the activation energy by handling the substrate molecule in such a way that the
reaction proceeds much more easily.

Enzymes stabilizes the transition state at that point when the enzymes are holding the substrate
molecules together form at that point is the transition state , so the enzyme stabilizes that transition state
allowing for bonds to form or bonds to break.

When a substrate binds to the active site of an enzyme, the shape of its molecule is slightly
changed which makes it easier for the substrate to change into a product thus the activation
energy is lowered.

diagram?
 CAPE BIOLOGY UNIT 1 ENZYMES TUTORIAL (PART 1)

8. Explain the two models for how substrates bind the active site of an enzyme. (4 mks)

The 'lock and key' model.

According to the lock and key model, the enzyme’s active site complements the substrate
precisely
The substrate fits a particular active site like a key fits into a particular lock
This theory of enzyme-substrate interaction explains how enzymes exhibit specificity for a
particular substrate

The 'induced fit' model.

According to the induced fit model, the enzyme’s active site is not a completely rigid fit for the
substrate
Instead, the active site will undergo a conformational change when exposed to a substrate to
improve binding
This theory of enzyme-substrate interactions has two advantages compared to the lock and key
model:
■ It explains how enzymes may exhibit broad specificity (e.g. lipase can bind to a variety of
lipids)
 CAPE BIOLOGY UNIT 1 ENZYMES TUTORIAL (PART 1)

■ It explains how catalysis may occur (the conformational change stresses bonds in the
substrate, increasing reactivity)

9. What is meant by (a) active site (3 mks);

There is a region on the enzyme that is a collector pocket that is specified at the tertiary level of
protein organization.A region on the surface of an enzyme whose shape permits binding
only of a specific molecular substrate that then undergoes catalysis.

-is the collector pocket in the enzyme where the substrate can bind.

The Active site is a cleft or pocket in the enzyme were substrates can bind
It has SPECIFICITY
1. Group specificity - Recognise functional groups (e.g. hexokinase; pepsin)
2. Absolute specificity (recognises a particular substrate e.g. glucokinase; lactase)

(b) enzyme specificity (2 mks

the extent to which an enzyme's activity is restricted to a specific substrate, a specific

group of substrate, a specific type of chemical bind, or a specific type of chemical
reaction.

Enzymes are specific because different enzymes have differently shaped active sites.The
shape of the active site of an enzyme is complementary to the shape of its specific
substrate. This means they are the correct shapes to fit together. Temperature has an
effect on enzyme activity.
 CAPE BIOLOGY UNIT 1 ENZYMES TUTORIAL (PART 1)

(c) denaturing (3 mks)

Denaturation of enzymes refers to the conformational changes in the protein structure and
active site of the enzyme and results in the inactivity of enzymes.

Denaturation is a process in which enzymes lose their conformational structure due to

application of external stress , excess heat or changes in pH.Most enzymes are folded into a
particular shape to function. H bonds ( hydrogen bonds )play an important role in protein
folding. H bonds are weak bonds that are easily altered by changes in temperature and pH.

Denaturation involves the breaking of many of the weak H bonds within an enzyme that are
responsible for the highly ordered structure of the enzyme. Most enzymes lose their activity once
denatured , because substrate can no longer bind to the active site.

(d) enzyme turnover number (2 mks)?

Enzymes are catalysts that speed up chemical reactions without getting used up in the
process. In reference to enzymes, turnover number means the maximum number of substrate
molecules that is being converted to product per second. It is represented by K cat
is 'The number of substrate molecules that a molecule of an enzyme converts into products
per second.
The number of substrate molecules converted into product in an enzyme-catalyzed
reaction under saturating conditions per unit time per unit quantity of enzyme; e.g., kcat =
Vmax/[Etotal].

10. Using a named example, explain the key stages in the mode of action of enzymes. (8 mks)

When the substrate molecule fits inside the active site which it is complementary to as a
result of enzyme specificity, the activation energy is lowered and the molecule is put
 CAPE BIOLOGY UNIT 1 ENZYMES TUTORIAL (PART 1)

under strain so that bonds break or form. The combination of enzyme and substrate is
an enzyme -substrate complex.

The enzyme and the substrate are in the same area. Some situations have more than one
substrate molecule that the enzyme will change.

2. The enzyme grabs on to the substrate at a special area called the active site. The
combination is called the enzyme/substrate complex. Enzymes are very, very specific and
don't just grab on to any molecule. The active site is a specially shaped area of the enzyme
that fits around the substrate. The active site is like the grasping claw of the robot on the
assembly line. It can only pick up one or two parts.

3. A process called catalysis happens. Catalysis is when the substrate is changed. It could
be broken down or combined with another molecule to make something new. It will break or
build chemical bonds. When done, you will have the enzyme/products complex.

4. The enzyme releases the product. When the enzyme lets go, it returns to its original
shape. It is then ready to work on another molecule of substrate.

Enzyme 1 Enzyme 2 Enzyme 3 Enzyme 4

11. A B C D E

If a cell was unable to make enzyme 3, what would happen in the cell to the level of: (a)
compound D; (b) compound C. (2 mks)

Recall that chemical reactions convert substrates into products, often by attaching
chemical groups to or breaking off chemical groups from the substrates.

12. Explain the effects of temperature, pH, enzyme concentration and substrate concentration on
enzyme action. Be sure to include expected graphs. (12 mks: 4 mks each)

Explain the effect of pH on enzyme activity.

 CAPE BIOLOGY UNIT 1 ENZYMES TUTORIAL (PART 1)

If pH is low, the concentration of hydrogen ions in the solution is high and this causes the -
COOH group to remain unionized. If pH is high, the concentration of hydrogen ions in the
solution is low and the -NH3 groups do not ionize.
Either of these conditions can therefore cause ionic bonds between R groups in the enzyme
molecule to break, thus denaturing the enzyme.

Explain the effect of temperature on enzyme activity.

As the temperature increases, the rate of reaction increases because there's an increase in kinetic
energy and molecules of the substrate collide more frequently with enzymes. However as the
temperature increases past the optimum temperature, the enzyme molecules vibrate and bonds
stabilizing the tertiary structure begin to break and are eventually denatured.

Explain the effect of enzyme concentration on enzyme activity.

As the concentration of enzymes increases, so does the rate of reaction. The rate of
reaction is directly proportional to the enzyme concentration, assuming that there is
excess substrate. Low substrate concentration is a limiting factor.

Explain the effect of substrate concentration on enzyme activity.

As substrate concentration increases, the initial rate of reaction also increases. The
more substrate molecules there are around, the more often an enzyme's active site can
bind with one. However enzyme concentration is a limiting factor.
 CAPE BIOLOGY UNIT 1 ENZYMES TUTORIAL (PART 1)

Enzymes tutorial (part 2)

1. Using named examples, distinguish between reversible and irreversible inhibition. (6 mks)

⚫ irreversible inhibition-organophosphates, Cox-2 inhibitors e.g. aspirin

⚫ reversible inhibition-edrophonium, physostigmine, and neostigmine
⚫

irreversible inhibition leaves a permanent effect on the temperature while reversible inhibition leaves a
temporary effect on the enzyme on enzyme activity.

The effect is permanent for irreversible inhibition and is temporary for reversible inhibition

The main difference between reversible and irreversible enzyme inhibition is that reversible
enzyme inhibition inactivates enzymes through noncovalent interactions. In contrast,
irreversible enzyme inhibition inactivates enzymes through covalent inactivation of the active
site. Furthermore, the inhibition effect is reversible in the reversible enzyme inhibition, but the
inhibition effect is irreversible in the irreversible enzyme inhibition.

2. Using a named example, explain how competitive inhibition affects enzyme activity. (5 mks)

Malonic acid is a competitive inhibitor. Its competes with succinate for the active sites of
succinic dehydrogenase, an important inside in the Krebs cycle.

Competitive inhibitors compete with the substrate for the active site of an enzyme molecule. The
inhibitor me have a structure which permits is to combine with the active site. While it remains
bound to the active site, it prevents substrate molecules from occupying that site and so reduces
the rate of reaction.

The competitive inhibitor binds to the active site and prevents the substrate from binding there.
The noncompetitive inhibitor binds to a different site on the enzyme; it doesn't block substrate
binding, but it causes other changes in the enzyme so that it can no longer catalyze the reaction
efficiently.

3. Using a named example, explain how non-competitive inhibition affects enzyme activity.

Examples of non-competitive inhibitors include cyanide and penicillin.

 CAPE BIOLOGY UNIT 1 ENZYMES TUTORIAL (PART 1)

Insecticides, example pyrethroids and nicotine inhibits the nervous system. Pyrethroids act on
the nerve axion and nicotine acts on the synapse of the central nervous system of the insects.

Non competitive inhibitors attach themselves not to the active site of the enzyme molecule but
elsewhere on the enzyme molecule. They alter the shape of the enzyme molecule in such a way
that the active site can no longer properly accommodate the substrate thus reducing the rate of
reaction by decreasing the enzyme concentrations.

Globular enzyme molecules are coiled into a precise three-dimensional dimensional shape, that is, their
teritiary structure, with hydrophilic R group ,that is, their side chains, on the outside of the molecules
making them soluble in water.