Biology - Form 4 - Track 3 Enzymes

Enzymes

What is an Enzyme? 2

How Enzymes Work 2

Enzyme Specificity 3

Effect of Temperature on Enzyme function and activity 3

Effect of pH on Enzyme function and activity 5

Enzyme Investigations 7
Investigating the Effect of Temperature on Amylase 7
Investigating the Effect of pH on Amylase Activity 8

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Biology - Form 4 - Track 3 Enzymes

What is an Enzyme?
Enzymes are biological catalysts which speed up the rate of a chemical
reaction without going under any change themselves. Enzymes are also huge
proteins.
Enzymes are necessary to all living organisms as they maintain reaction
speeds of all metabolic reactions (all reactions which keep an organism alive),
including respiration.

How Enzymes Work

● Enzymes and substrates move randomly in a solution.
● When an enzyme (active site) and its complementary substrate collide
an enzyme-substrate complex forms, and the reaction occurs.
● Product/s forms from the substrate which are then released from the
active site.
● The enzyme is unchanged and will go on to catalyze further reactions.

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Biology - Form 4 - Track 3 Enzymes

Enzyme Specificity
Enzymes are specific to one particular substrate as the active site where the
substrate attaches is a complementary shape to the substrate. This is due to
the enzyme being a protein and having a specific shape.
This means that only a certain enzyme can form an enzyme-substrate
complex with a certain substrate. This is known as the lock and key
mechanism.

Effect of Temperature on Enzyme function and activity

Since enzymes are proteins and have a specific shape which is held by bonds, it
is important that their shape is maintained so the substrate will fit into the
active site and catalyze the reaction.
Enzymes work faster at higher temperatures, especially ones which are closer
to their optimum pH, this is because since more kinetic energy is present, the
substrate will collide more often with the active site (increase collisions),
speeding up the reaction.

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Biology - Form 4 - Track 3 Enzymes

Heating an enzyme to high temperatures (beyond their optimum) will break

the bonds that hold the enzyme together. This will denature the enzyme,
stopping the reaction as the substrate won't be able to fit into the active site
as its shape and active site will be changed.

If an enzyme is exposed to low temperature, their kinetic energy decreases

meaning they collide less often with the active site and the reaction slows
down. This does not denature the enzyme, as if it is heated up the reaction will
occur at a faster rate.

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Biology - Form 4 - Track 3 Enzymes

Effect of pH on Enzyme function and activity

All enzymes have an optimum pH at which they function best. The optimum
pH for most enzymes is 7, but some that are produced in acidic condition, such
as the stomach, have a lower optimum pH (pH 2) and some that are produced
in alkaline conditions, such as the duodenum, have a higher optimum pH.

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Biology - Form 4 - Track 3 Enzymes

If the pH is too high or low (depending on the enzyme), the bonds that hold the
amino acid chain together make up the protein can be destroyed. This will
change the shape of the active site, so the substrate can no longer fit into it,
reducing the rate of activity.

Moving too far away from the optimum pH will cause the enzyme to denature
and all activity will stop.

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Biology - Form 4 - Track 3 Enzymes

Enzyme Investigations

Investigating the Effect of Temperature on Amylase

Method:

● Starch solution is heated to a set temperature.

● Iodine is added to wells of a spotting tile.
● Amylase is added to the starch solution and mixed well.
● Every minute, droplets of solution are added to a new well of iodine
solution.
● This is continued until the iodine stops turning blue-black (this means
there is no more starch left in the solution as the amylase has broken it
all down).
● Time taken for the reaction to be completed is recorded.
● Experiment is repeated at different temperatures.

Conclusion: The quicker the reaction is completed, the faster the enzyme is
working, meaning the greater its kinetic energy is as they are colliding more
with the active site.

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Biology - Form 4 - Track 3 Enzymes

Investigating the Effect of pH on Amylase Activity

Method:

● Place single drops of iodine solution in rows on the tile

● Label a test tube with the pH to be tested
● Use the syringe to place 2cm3 of amylase in the test tube
● Add 1cm3 of buffer solution to the test tube using a syringe
● Use another test tube to add 2cm3 of starch solution to the amylase and
buffer solution, start the stopwatch whilst mixing using a pipette
● After 10 seconds, use a pipette to place one drop of mixture on the first
drop of iodine, which should turn blue-black
● Wait another 10 seconds and place another drop of mixture on the
second drop of iodine
● Repeat every 10 seconds until iodine solution remains orange-brown
● Repeat experiment at different pH values

Conclusion: The less time the iodine solution takes to remain orange-brown,
the quicker all the starch has been digested and so the better the enzyme
works at that pH.

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Biology - Form 4 - Track 3 Enzymes

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