Temasek Junior College

IP3 Biology Think Cycle

 Know
Topic: Enzymes
Notes
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Learning Outcomes
Candidates should be able to:
(a) Explain the mode of action of enzymes in terms of an active site, enzyme-substrate
complex, lowering of activation energy and enzyme specificity using the lock-and-key
and induced-fit hypotheses. [O and A-levels]
(b) Investigate and explain the effects (with reference to protein structure and shape of
active site) of temperature, pH, enzyme concentration and substrate concentration on
the rate of enzyme catalysed reactions and explain these effects. [O and A-levels]

Use the knowledge gained in this section in new situations or to solve related problems.

1. INTRODUCTION
2. MODE OF ACTION OF ENZYMES
3. PROPERTIES OF ENZYMES
4. FACTORS AFFECTING RATE OF ENZYME ACTION

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I. INTRODUCTION TO ENZYMES

 Enzymes are protein molecules which act as biological catalysts.

 They greatly increase the rate of a chemical reaction,

 without themselves being changed at the end of the reaction.

 Without enzymes, biochemical reactions would proceed too slowly to sustain life.
Raising the temperature could increase the speed of reaction, but this would be
detrimental to the cell.

 Enzymes enable metabolic reactions to proceed rapidly but at low temperatures.

 Enzymes are specific. A single enzyme will generally catalyse only a single reaction.

 Enzymes serve to control the reactions within each metabolic pathway.

II. PROPERTIES OF ENZYMES

 Most enzymes:
I. are globular proteins.
II. have active sites with a specific shape where the reaction takes place.

 Enzymes function as biological catalysts and hence share the properties of catalysts.
 They are effective in small amounts.
 They remain chemically unaltered at the end of the reaction they catalyse.
 They function by lowering the activation energy required for the reaction to occur.

 Enzymes are extremely efficient.

 Most enzyme-catalysed reactions are highly efficient, proceeding 103 to 108 times
faster than uncatalysed reactions.

 Enzymes have a high degree of specificity.

- Most enzymes are specific to one type of substrate molecule. For example, catalase
will only catalyse the decomposition of hydrogen peroxide.
- Some enzymes are specific to a group of substrates which have similar shapes.
- The specificity of an enzyme is due to the conformation at its active site.

 The activity of enzymes is affected by changes in: (Covered in Section IV).

I. pH,
II. temperature,
III. substrate concentration
IV.enzyme concentration.

 The activity of enzymes can also be changed by the presence of inhibitors or

activators.
 This implies that the rate of product formation can be controlled according to the
needs of the cell.

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III. THE MODE OF ACTION OF ENZYMES LO (a)

(a) General Structure of An Enzyme Molecule

- All enzymes have a complex globular shape and are relatively large molecules (far
larger than the substrates that they act on).

- Despite its size, only the active site of the enzyme molecule comes into direct
contact with the substrate.

- A substrate binds to the active site of the enzyme, forming the enzyme-substrate
complex.

- The shape of the active site is complementary to the shape of the substrate. (Fig 1)

Fig. 1

(b) Activation Energy (Ea)

- Definition: Activation energy is the energy required to make substances react.

- It represents the energy barrier that has to be overcome before a reaction can take
place to form products. (Fig. 2)

- The larger the Ea, the slower will be the reaction at any particular temperature.

- If the Ea of a reaction is lowered, the rate of reaction would be increased. (Fig. 2)

- Enzymes are special biological catalysts, which serve to reduce the activation
energy required for a chemical reaction to start.

For your information:

 The activation energy of a reaction can be supplied by heating the reactants – the
reaction will then proceed at a faster rate.
 However, this is not possible in the living cell. Cells only survive within a narrow
range of relatively low temperatures – heating will often kill the cell.
 Hence the need for enzymes 

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 Fig 2: Activation energy for an enzyme-catalysed and an uncatalysed reaction

(c) How Enzymes Work

- TWO hypotheses have been suggested to describe the mechanism of enzyme

action:

1. The Lock-and-Key Hypothesis (Proposed by Fischer in 1890) (Fig. 3)

I. The active site has a specific shape, to which the substrate binds.

II. The substrate is imagined to be like a key, whose shape is complementary

to the shape of the enzyme active site (or the lock).

III. The shape of substrate or substrates fit exactly into the shape of active site.
IV. Once the reaction is completed with products formed, the products no longer
fit into the active site, and are released into the surrounding medium, leaving
the active site free to receive new substrate molecules.

Fig. 3: The “lock-and-key” hypothesis of enzyme action

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2. The Induced Fit Hypothesis (Proposed by Koshland in 1959) (Fig. 4)

I. This stemmed from evidence that suggested that some enzymes and their
active sites were physically flexible structures.

II. Shape of substrate is still complementary to shape of active site but does
not fit exactly.

III. Binding of the substrate to the active site induces a small conformational
change in the shape of the enzyme.

 This enables the substrate to fit more snugly into the active site when
they form the enzyme-substrate complex

 This allows the enzyme to perform its catalytic function more effectively.

Fig. 4: The induced-fit hypothesis of enzyme action

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  This is how enzymes work at the molecular level:

Collision between substrate and enzyme at the correct orientation

causes the substrate to bind to the enzyme molecule at its active site,
to form a short-lived enzyme-substrate complex. Within the complex,
the chances of reactions occurring are greatly enhanced.

The reaction occurs. Enzymes are

reused

Once reaction has occurred, the products are formed and

subsequently released.
The unchanged enzyme molecule is also released.
This is then available to catalyse another cycle of reaction.

enzyme + substrate

enzyme-substrate
complex

enzyme + products

Fig. 5: A diagrammatic representation of an enzyme-catalyzed reaction

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IV. FACTORS AFFECTING RATE OF ENZYME ACTION LO (c)

A. Effects of temperature
B. Effects of pH
C. Effects of substrate concentration
D. Effects of enzyme concentration

A. EFFECTS OF TEMPERATURE

 As temperature increases, there is an increase in kinetic energy of enzyme and

substrate molecules.

 This results in an increase in the number of effective collisions between enzyme and
substrate molecules.
 more enzyme-substrate complexes are formed per unit time. (Fig. 6)

 The rate of reaction increases with temperature until the optimum temperature is
reached. (Fig. 6)

 If the temperature is increased beyond the optimum temperature, the rate of

enzymatic reaction decreases rapidly. This is because the enzyme is denatured. (Fig.
6)

 Optimum temperature is the temperature at which the enzyme is functioning at

its maximum rate.

 The optimum temperature of most mammalian enzymes lie between 30 – 40 C, but
enzymes with higher optimum temperatures exist.

What happens when an enzyme is denatured:

a. Since enzymes are made of proteins, excessive heat disrupts the intra-molecular
bonds (especially the hydrogen bonds and hydrophobic interactions) which
stabilize the (tertiary and quaternary) structures of the enzyme.
b. The enzyme unfolds and the precise shape of the active site is lost. The
enzyme is said to be denatured.
c. Enzyme denaturation is usually irreversible.
What happens when an enzyme is placed at freeing point:
 If the temperature is reduced to near or below freezing point, the enzymes are
inactivated. Enzyme activity is very low (lesser kinetic energy), but they will regain
their catalytic influence when higher temperatures are restored.

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 Fig. 6: The effect of temperature on the rate of an enzyme-controlled reaction

B. EFFECTS OF pH

1. Under conditions of constant temperature, every enzyme functions most particularly

over a specific range of pH.

2. The optimum pH of an enzyme is the pH at which maximum rate of reaction

occurs.

a. At this pH, the intramolecular bonds which maintain the protein structure of the
enzyme are intact. (Fig. 7)

b. The conformation of the active site is most ideal for substrate binding. (Fig. 7)

c. Hence, there will be highest number of effective collision between enzyme and
substrate molecules resulting in the greatest number of enzyme-substrate
complexes formed per unit time. (Fig. 7)

3. At pH higher or lower than the optimum pH, the H+ concentration would have
changed. (Fig. 7)

a. This alters the bonds (hydrogen bonds) that help to maintain the conformation of
the enzyme
 active site and substrate binding would be affected. (Fig. 7)

4. Within a certain range, the effects of pH are normally reversible. Restoring pH to the
optimum would usually restore the optimum rate of reaction. (Fig. 7)

5. At extreme pH, the conformation of the enzyme is severely altered. It is hence

denatured.

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 Points 2a-c)

Point 5

Figure 7: The effect of pH on the rate of reaction of two digestive enzymes

C. EFFECTS OF SUBSTRATE CONCENTRATION

1. For a fixed enzyme concentration, the rate of enzyme reaction increases with
increasing substrate concentration. (Fig. 8)

a. An increase in number of substrate molecules leads to an increase in the

frequency of effective collisions between enzyme and substrate molecules.
 more enzyme-substrate complexes formed per unit time.
 rate of reaction is increased.

b. The rate of reaction is limited by substrate concentration. (Fig. 8)

2. There comes a point when any further increase in substrate concentration produces
no significant change in rate of reaction. (Fig. 8)

a. This is because all the active sites of all the enzyme molecules are saturated with
substrate molecules.

b. Any extra free substrate molecule has to wait until the enzyme-substrate complex
has released the products before it may itself enter the active site of the enzyme.

c. The rate of reaction is now limited by enzyme concentration.

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 Fig. 8: Effect of substrate concentration Fig. 9: Relationship between enzyme
concentration and the rate of an
enzyme-controlled reaction

D. EFFECTS OF ENZYME CONCENTRATION

1. Provided that the substrate concentration is maintained at a high level, and other
conditions such as pH and temperature are kept constant, the rate of reaction is
proportional to the enzyme concentration. (Fig. 9)

a. As enzyme concentration is increased, the frequency of effective collision

between enzyme and substrate molecules increases.

b. Greater number of enzyme-substrate complexes is formed per unit time.

2. At very high enzyme concentrations, if the concentration of substrate molecules is

limiting, the increase in enzyme concentration would not result in any further increase
in rate of reaction. (Fig. 9)

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