UNIT 5: ENZYMES

Many chemical reactions can be speeded up by substances called

catalysts. A catalyst is a substance that increases the rate of a
chemical reaction and is not changed by the reaction. Within
living organisms, these reactions (metabolic reactions) are controlled
by catalysts called enzymes. Enzyme molecules are proteins.
Definition: An enzyme is a protein that is involved in all
metabolic reactions, where it functions as biological catalyst.
An enzyme-controlled reaction involves three groups of molecules,
although the product may be two or more different molecules.
Substrate enzyme products

The substance on which an enzyme acts is called its substrate and

the molecules produced are called the products.
E.g.: The enzyme sucrase acts on the substrate sucrose to produce
monosaccharide products glucose and fructose.
At any moment in our body, many chemical reactions are taking
place, building new molecules, breaking others down and changing
others. All these reactions are catalysed by enzymes. Most of these
reactions could happen without enzymes but would go too slowly
for the organisms to survive. Catalase, a very common enzyme
found in most cells, can break down 40 000 molecules of hydrogen
peroxide every second! A complete chemical reaction takes only a
few seconds when the right enzyme is present.
Enzymes are named by adding the suffix -ase to the name of the
substrate that they modify (i.e., maltase, lipids), or the type of
reaction they catalyze (dehydrogenase, decarboxylase).

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Properties of enzymes:
1. Proteinaceous nature: Nearly all enzymes are proteins.
2. Substrate specificity: A given enzyme only catalyzes one
reaction or a similar type of reaction. For example, maltase
acts only on maltose while pancreatic lipase acts in a
variety of fats. Sometimes, different enzymes may act on
the same substrate to produce different end products. The
substrate specificity of enzyme is based on amino acids
sequence in the active site.
3. Catalytic properties
(i) Enzyme require in small concentration for any
chemical change.
(ii) They don’t initiate the catalysis but accelerate the
rate of catalysis by lowering the activation energy.
(iii) They remain unchanged at the end of reaction.
(iv) Their presence doesn’t alter the properties of end
products.
4. Sensitivity:
Enzymes are highly sensitive to change in pH and
temperature. Enzymes work best at a narrow range of
condition called optimum level.
5. Inhibitors: Enzymes are also sensitive to inhibitors.
Inhibitors are any molecules like cellular metabolites,
drugs or toxins which reduce or stop enzyme activity.
Enzymes - 'Lock and key' model
Enzymes are very specific; each kind of enzyme catalyse one kind
of reaction only. To catalyse a reaction, enzyme molecule and
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substrate molecule need to meet and join together by a temporary
bond. The shape of the active site of the enzyme molecule and the
substrate molecule are complementary.

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Temperature, pH and enzymes
The activity of enzymes is affected by temperature and pH.

Effect of temperature on enzymes (graph to be drawn.)

The optimum (best) temperature for enzyme-controlled reactions is
37oC (body temperature). As the temperature increases, the rate of
reaction increases. But very high temperatures denature enzymes.
The graph shows the typical change in an enzyme's activity with
increasing temperature. The enzyme activity gradually increases
with temperature up to around 37ºC, or body temperature. Then, as
the temperature continues to rise, the rate of reaction falls rapidly
as heat energy denatures the enzyme. Most enzymes are denatured
above 50oC.

Effect of pH on enzymes (Graph to be drawn)

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The pH of a solution is how acidic or alkaline it is. Acid or alkaline
conditions alter the chemical properties of proteins, including
enzymes.
Different enzymes work best at different pH values. The optimum
pH for an enzyme depends on where it normally works. It is around
neutral (pH= 7) for most enzymes but there are some exceptions.

Changes in pH alter an enzyme’s shape and slow down its activity,

but this can usually be reversed if the optimum pH is restored. An
extreme pH can denature enzymes – the active site is deformed
permanently.

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Rates of enzyme reactions:
The rate of enzyme-controlled reaction depends on the temperature
and pH. It also depends on the concentration of the enzyme and its
substrate. The more enzyme molecules produced by a cell, the faster
the reaction will proceed, provided there are enough substrate
molecules available. Similarly, an increase in the substrate
concentration will speed up the reaction if there are enough enzyme
molecules to cope up with the additional substrate.
Investigations

1. The effect of temperature on an enzyme reaction.

• Draw 5 cm3 of amylase in a syringe and place 1 cm3 in each of 3 test tubes labelled A, B
and C.
• Place 5 cm3 of 1% starch solution in each of test tubes labelled 1, 2 and 3.
• To each of test tubes 1, 2 and 3, add 6 drops of iodine solution using a dropper.
• Prepare 3 water baths: one with ice cold water of about 10 oC, second water bath with
35 oC and the third water with 50 oC.
• Place tubes 1 and A in the cold-water bath. Tubes 2 and B in second water bath and tubes
3 and C in third water bath.
• Incubate them for 5-10 min in respective water bath.
• After 10 min, take the temperature of each water bath and pour the amylase from tube A
into starch solution in tube 1 and return tube 1 to the water bath.
• Repeat this with tubes 2 and B, and 3 and C.
• As the amylase breaks down starch, it will cause the blue colour to disappear. Time taken
for disappearance of colour in all the tubes should be noted.

Note: Second investigation should be referred from the Textbook

2. The effect of pH on an enzyme reaction

• Label 5 test tubes 1-5 and place 5 cm3 of 1% starch solution in each tube.
• Add acid or alkali to each test tube to adjust the pH to 10, 8, 7, 5 and 3 respectively in 5
tubes. Alternatively, buffer solutions can be used.
• Place several rows of iodine drops in cavity tile.
• Place 1 cm3 of 5% amylase solution in each tube. Shake the contents and note the time.

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 • Use a dropping pipette to remove a small sample from each tube and transfer to one of
the iodine drop on the cavity tile. Keep on sampling this way.
• When any of the samples fail to give a blue colour, this means that the starch in that tube
has been completely broken down to sugar by the amylase. Note the time when this
happens for each tube and stop taking samples from that tube. Do not continue sampling
for more than 15 min.

Note: Second investigation should be referred from the Textbookss