Effect of temperature on amylase

Aim: To investigate the effect of temperature on the breakdown of starch by amylase.

Equipment
 Test tube rack with 2 test tubes  Waterbaths;
 Spotting tile 5 oC, 25 oC, 45oC, 65oC, 85 oC
 Marker pen  Starch solution
 Stopwatch  Amylase solution
 Iodine  Glass rod
 Thermometer

Method
1. Add two drops of iodine solution to each well in a spotting tile
2. Label two test tubes with your initials.
3. Label one test tube with E and the other with S.
4. Add 2cm3 of amylase to the E test tube and 2cm3 of starch solution to the S test
tube.
5. Place the test tubes in the appropriate water bath for 5 minutes.
6. Pour the contents of the E tube into the S tube and start the stopwatch.
7. Immediately remove a sample from the test tube and place into the first well of the
spotting tile using the glass rod and stir into the iodine.
8. Rinse and dry the glass rod.
9. Each 30 seconds remove another sample from the tube and place into the next well.
10. You may need to rinse and reset the spotting tile.
11. Record the results in the table.
12. Repeat steps 4 to 11 with the other four temperatures.

Test Temperature Time taken for yellow colour Rate of reaction

tube (oC) to remain (s) (1/time)
1 5 850

2 25 600

3 45 450

4 65 840

5 85 920

Analysis
Calculate 1/time to give the rate of reaction and complete the column in the table.
Plot a graph of temperature (oC) against rate of reaction (1/time)

Conclusion
Using your data and research you find elsewhere (cited appropriately), draw conclusions
from your data. Think about the kind of reaction that amylase catalyses, where you find it in
the body and how this links to the temperature at which it works best.
Evaluation
How could this investigation be made; more accurate, more reliable? How could this
investigation be extended?