ENZYMES

LEARNING OUTCOMES
Candidates should be able to:
a. define catalyst as a substance that speeds up a chemical reaction and is not changed
by the reaction
b. define enzymes as proteins that function as biological catalysts
c. explain enzyme action in terms of the ‘lock and key’ hypothesis
d. investigate and describe the effects of temperature and of pH on enzyme activity .

A catalyst is a substance which speeds up the rate of a reaction without itself being
changed at the end of the reaction.
 All chemical reactions taking place inside the body are known as metabolism.
 Without enzymes the reactions would be so slow that life cannot be
maintained.
 Enzymes are biological catalysts which speed up the rate of metabolism.
 Enzymes occur inside cells or are secreted by the cells.
 Catalase is the enzyme that catalyses the break down of hydrogen peroxide.

The properties of enzyme

1. All enzymes are proteins- This may seem rather odd, because some enzymes actually
digest proteins.
2. Enzymes are made inactive by high temperature- This is because they are protein
molecules, which are damaged by heat.
3. Enzymes work best at a particular temperature- Enzymes which are found in the
human body usually work best at about 37 °C.
4. Enzymes work best at a particular pH- pH is a measure of how acid or alkaline a
solution is. Some enzymes work best in acid conditions (low pH). Others work best in
alkaline conditions (high pH).
5. Enzymes are catalysts- They are not changed in the chemical reactions which they
control. They can be used over and over again, so a small amount of enzyme can
change a lot of substrate into a lot of product.
6. Enzymes are specific- This means that each kind of enzyme will only catalyse one kind
of chemical reaction.

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 How do enzymes work?
 Enzymes are thought to have an area with a very particular shape.

 When a molecule of the right chemical for that enzyme comes along it will fit
exactly into the shape.

 The area of particular shape is called the active site of the enzyme, as that is
where the reaction takes place.

 The molecule that the enzyme works on is called the substrate. Substrates
when broken down are transformed into products which leave the active site
ready for another reaction.

 The active site of an enzyme has such a particular shape that only one kind of
molecule will fit it.

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 Lock and key hypothesis

 Just like a key fits into a particular lock in the same way a substrate fits into
the active site of an enzyme.

 This is known as the lock and key hypothesis.

 LEKS

 L=Lock is Enzyme

 K= Key is Substrate.

Examples of enzymes

Enzymes are commonly named by “-ase” ending.

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 Factors affecting rate of enzyme reaction

There are 2 factors affecting the rate of enzyme action.

These are:

1. Temperature and

2. pH

Temperature

 At low temperatures enzyme controlled reactions go slowly because the

molecules have low kinetic energy.

 Increasing the temperature increases the kinetic energy of the enzyme

and substrate molecules so that they move faster and are more likely to
collide.

 So increasing the temperature increases the rate of the reaction up to a

certain temperature.

 This temperature is known as the enzyme’s optimum temperature.

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 Different enzymes have different optimum temperatures. The enzymes in
animal bodies work best at 37˚C.

 If the temperature is increased beyond the optimum the enzyme has so

much kinetic energy that the bonds holding the enzyme molecule together
start to vibrate and eventually break.

 The enzyme loses its specific shape so that the substrate no longer fits in to
the active site. We say that the enzyme is denatured.

 Denaturation by temperature is irreversible.

 The enzyme changes shape and the active site no longer matches the
shape of the substrate molecule.

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 pH

 Enzymes prefer to work at an optimum pH. Outside of its pH range the

enzyme is denatured. Denaturation by pH is reversible.

 The pH of a solution affects the shape of an enzyme.

 Most enzymes have their correct shape at a pH of about 7 – that is, neutral.

 If the pH becomes very acidic or very alkaline, then they lose their shape.

 This means that the active site no longer fits the substrate, so the enzyme
can no longer catalyse its reaction.

Enzyme Optimum pH
Lipase 8.0
Pepsin 1.5 - 1.6
Trypsin 7.8 - 8.7
Urease 7.0
Invertase 4.5
Maltase 6.1 - 6.8
Amylase 6.7 - 7.0
Catalase 7.0

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 Experiment to investigate effect of temperature on enzyme action

The enzyme amylase, which is contained in your saliva, breaks down starch into sugar. The
presence of starch can be tested using iodine solution. In the presence of starch iodine turns
blue/black and if no starch is present it is a pale yellowy-brown colour. Iodine can therefore be
used to monitor the breakdown of starch into sugar. When the iodine no longer changes colour,
you know all of the starch has been broken down.

1. Test tubes containing 5 cm3 of starch were pre-warmed in a water bath at the appropriate
temperature for 5 minutes.
2. 1 cm3 of amylase was also placed in each water bath in a separate test tube.
3. After 5 minutes, when the starch and amylase had reached the temperature at which we were
working, the two tubes were mixed together and a stopwatch started.
4. The tube containing the reaction mixture was then kept in the waterbath at the appropriate
temperature.
5. After one minute a few drops of the reaction mixture were added to iodine on a spotting tile to test
for the presence of starch.
6. When the iodine no longer changed colour (i.e. no starch present) the time was recorded. The
results of the experiment are shown below.

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The results show that the reaction was most rapid at 40 oC and the starch remained undigested at
10oC and 60oC after 12 minutes. These results can be plotted on a graph to show how the rate of
reaction varies with temperature:

Note that the rate of reaction increases from 10oC to 40oC. Increasing the temperature causes the
reactants to move faster making a collision between the enzyme and the substrate more likely. At
40oC the rate of reaction reaches a maximum. Above this temperature, the enzyme becomes
denatured and it can no longer catalyse (speed up) the reaction.

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Experiment to investigate the effect of pH on the rate of enzyme catalysed
reaction

 1 cm3 of amylase solution and 5 cm3 of starch suspension are placed in separate test-tubes.

 2 cm3 of buffer* solution at pH 2 is placed in tube containing amylase.

*a buffer solution is one which resists changes in pH and keeps pH constant

 Both tubes are placed in water bath at 37 oC for 3 minutes.

 After 3 minutes, amylase with buffer solution are poured into the tube containing starch.

 A stopwatch is started immediately.

 Mean while drops of iodine are placed on a tile.

 At intervals of 30 seconds, 2 drops of mixture from the tube are placed in the iodine drops until the
solution remains pale brown or pale blue.

 The experiment is repeated using buffers at different pH such as 4, 6, 8, 10, 12 etc.

 The time taken at each buffer is tabulated and a graph plotted.