2.

5 Enzymes

Enzymes’ specificity

Enzymes:

- 3-D globular proteins

- act as catalysts; speed up chemical reactions.
- Has an active site to which the specific substrate binds.
- Enzyme catalysis involves molecular motion and the collision of substrates with the
active site.

Active site: It is the region on an enzyme to which the specific substrate binds

- A specific substrate fits the active site; the shape of the substrate corresponds to
the shape of the active site.
- The bonds in the substrate (s) are weakened making a reaction easier.
- The unchanged enzyme and product(s) are released.
- The enzyme is to be reused.

Lock and key hypothesis

The enzyme-substrate complex can be compared to a lock and key, where the enzyme is the
lock and the substrate is the key.

Substrate + enzyme enzyme-substrate complex product + enzyme

- A specific substrate fits the active site; the shape of the substrate corresponds to
the shape of the active site.
- The bonds in the substrate (s) are weakened making a reaction easier.
- The unchanged enzyme and product(s) are released.
- The enzyme is to be reused.

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The induced-fit model

 The substrate is specific to the active site of the enzyme

 The substrate does not fit perfectly into the active site of the enzyme.
 When the substrate binds to the active site, this changes the shape of the active site
and only then it perfectly fits the substrate. The substrate induces the active site to
change slightly.

 This weakens the bonds in the substrate and therefore reduces the activation
energy.
 This model is a more precise version of the lock and key one as it explains why some
enzymes can bind to many different substrates. That is why some enzymes can have
broad specificity.

Temperature, pH and substrate concentration affect the rate of activity of enzymes.

The effect of temperature on enzyme activity.

(a) As the temp increases, the kinetic energy of the

substrate and enzyme molecules increases.
Therefore, more collisions between the substrate
and the enzyme which causes the rate of reaction
to increase.

(b) At the optimum temp, the max rate of reaction.

(c) Beyond the optimum temp, the enzyme begins to

lose its 3-D shape as the bonds start to breakdown.
When the enzyme loses its shape including the
shape of its active site, it is denatured and the
substrate no longer fits the shape of the enzyme.

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 Note:

pH is dependent on the relative no.

The effect of pH on enzyme
activity of (H+) compared to (OH-)

Acids: give off H+, pH ˂ 7 , ex HCl

Bases: give off OH-, pH ˃ 7, ex NaOH

 There is no one optimal pH for all enzymes.

 At the optimal pH (a) or (b), the enzyme works most efficiently and the max rate of reaction is
reached.
 Some specialized enzymes have optimal pH that is acidic. E.g pepsin, optimum pH (a) = 1.5
 Most enzymes in the human body have an optimum pH near to 7. E.g salivary amylase,
optimum pH (b) = 7.2
 Above or below the optimal pH, the rate of reaction decreases.
 Too acidic or too basic will result in an enzyme becoming less efficient and inactive. If too many
positive or negative charges, the ionic bonds are disturbed, 3D shape of the enzyme alters and
the enzyme denatures.

The effect of substrate concentration on enzyme activity

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 (a) As the substrate concentration increases, the
more collisions between the substrate and
(b) enzyme. This will increase the rate of reaction.

(b) At a certain point, further increase in the

substrate concentration will no longer increase
the rate of reaction. This is because enzymes’
active sites become fully saturated with
substrates and therefore no longer active sites
are available. The rate becomes constant.

The use of lactase in the production of lactose-free milk

Lactase
Lactose + water glucose + galactose

Lactose:

 sugar of milk
 a disaccharide
 can’t be absorbed by the gut.
 Lactase is the enzyme that hydrolyses lactose into the monosaccharides glucose and
galactose are easily absorbed by the gut.

Lactose intolerance:

Many adults lack the enzyme lactase and so cannot break down lactose leading to
lactose intolerance; as people get older, they lose their ability to produce lactase. Milk
and milk products enter the digestive tract and are not digested; instead bacteria in the
large intestine feed directly on lactose resulting in nausea, abdominal pain, diarrhea,
cramps and excessive gas.

Production of lactose-free milk:

- Milk and milk products can be treated with lactase before consumption.
- Lactase can be obtained from fungi.
- Lactose-free milk can be made in two ways:
1. adding lactase directly to the milk so that the milk contains the enzyme.
2. immobilizing the enzyme in alginate beads and carry out experiments in which
the lactose in milk is hydrolysed. Immobilized enzymes are widely used in
industry.

- Production of lactose-free milk is an example of an industrial process depending on

biotechnology methods which are of huge and increasing economic importance.

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Advantages for producing lactose-free milk:

1. it allows people who are lactose intolerant to consume milk

2. galactose and glucose taste sweeter than lactose, so this reduce the need for
additional sweetener
3. galactose and glucose are more soluble than lactose / gives smoother texture /
reduces crystalization in ice cream
4. Bacteria ferment glucose and galactose more rapidly than lactose, shortening
production time of yogurt.

 Development of some techniques benefits particular human populations more than

others.
For example, the development of lactose-free milk available in Europe and North
America would have greater benefit in Africa/ Asia where lactose intolerance is more
prevalent. The development of techniques requires financial investment.

Practical 3:

Experimental investigation of
a factor affecting enzyme
activity.