Date:

Topic: enzyme

Aim: Investigating the effect of temperature on the enzyme amylase

Materials:

● test tubes

● a test tube rack

● water baths (electrical or Bunsen burners and beakers)

● spotting tiles

● a 5cm measuring cylinder or syringe

3

● glass rods

● a stopwatch 

● starch solution

● amylase solution

● iodine solution 

● Thermometers.

Method:

● One drop of iodine solution was placed into each depression on the spotting tile.

● Water baths were set up for every temperature that was tested. (one cold with ice, one at room
temperature, one around body temperature 35 0c - 400c and one above 500c).
● 5cm3 of starch solution was measured using the measuring cylinder, into 4 test tubes.

● One test tube of starch solution was placed into each water bath.

● 1cm3 of amylase solution was measured out using the measuring cylinder, into 4 different test
tubes.
● One test tube of amylase solution was placed into each water bath.
 ● Test tubes were left in the water baths until the contents of each test tube reached the
temperature of the water bath.
● A note of each test tube temperature was taken. Then the amylase solution was carefully
poured into the test tube of starch solution and was mixed with a glass rod.
● One drop of the mixed solution was removed from the end of the glass rod and was placed on
the first depression of the spotting tile with iodine solution. This was ‘time zero’.

● The stop clock was immediately started.

● A glass rod was used to remove one drop every minute and place onto the iodine solution in the
next depression on the spotting tile. Glass rod was rinsed with water after each drop.
● Continued step11 until the iodine solution was no longer turning black. That indicates that the
starch has been broken down.
● Temperature of the water bath was recorded and how long the starch took to break down.

● This was repeated for other temperature.

Result/ observation:

Temperature of water bath

in °C
Time taken for amylase to completely break down the

starch in minutes

00c 4mins
200c 2m 30s

370c 1min

600c 0

Discussion: An enzyme is a substance produced by a living organism which acts as a catalyst to bring
about a specific biochemical reaction. Enzymes speed up the rate of a chemical reaction to help support
life. The optimum temperature for enzymatic activity of salivary amylase ranges from 32 0C to 370C. The
optimum temperature means that the temperature at which the enzyme shows the maximum activity.
At the optimum temperature the amylase will break down starch very quickly. This happens
because, while higher temperatures do increase the activity of enzymes and the rate of
reactions, enzymes are still proteins, and as with all proteins, temperatures above 37 degrees
Celsius, will start to break them down. This shows in the table above, when it was at 370c. At
low temperatures the amylase will break starch down slowly due to reduced kinetic energy. This
means that the enzyme is inactive; it shows in the above at 00c. At high temperatures the amylase
will break starch down slowly or not at all due to denaturation of the enzyme’s active site. This
means that the high temperatures destroy amylase.If the enzyme is exposed to high temperatures
the shape of the active site may change. When this happens the substrate will no longer fit into
the enzyme. Enzymes bind with chemical reactants called substrates. The enzyme’s active site
binds to the substrate, so when temperatures are high the enzyme's active site shape changes
causing the active site and the substrate to not bind.

Conclusion: it can be concluded that at high and low temperatures the shape of the enzyme
amylase is changed.