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Effect of temperature on enzyme activity Problem statement What is the effect of temperature on the activity of the salivary amylase on starch? Hypothesis As the temperature increases, the rate of the enzyme reaction increases until it reaches the maximum rate at an optimum temperature of 37 °C. Then it decreases and stops at 60 °C. Variables

a) Manipulated:

b) Responding:

c) Constant:

Temperature of the medium

Rate of enzyme reaction, time taken for the iodine solution to remain brown (time taken for the starch to be hydrolise)

The concentration of the enzyme amylase, the concentration of starch solution

Apparatus and materials Thermometer, beaker, test tubes, white tiles, water bath, stopwatch, distilled water, dropper, 1 % starch solution, iodine solution, measuring cylinder, saliva Technique Measure and record the time taken for the iodine solution to remain brown/Measure and record time taken for ,he starch to be hydrolise

and record time taken for ,he starch to be hydrolise Procedure 1 2 3 4 5

Procedure

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3 ml of saliva is collected in a beaker and then diluted with 3 ml of distilled water.

1 ml of the diluted saliva is placed in each test tube labelled P, Q, R, S and T.

5 mi of 1 % starch solution is placed in another set of test tubes labelled P1,

The test tubes labelled P and P1 are placed in the water bath at 5 °C for five minutes.

Meanwhile, many droplets of iodine solution are prepared on the white tile (Figure

4.15).

After 5 minutes, the saliva in test tube P is poured into test tube P1. The mixture is stirred and the stopwatch is started immediately.

A drop of the mixture is placed on a drop of iodine solution. At intervals of 1 minute, a drop of the mixture is placed on one of the drops of iodine solution on the white tile.

When the mixture shows no further positive reaction with the iodine solution, i.e. no blue-black colour, the time taken for this to happen is recorded. This is the time taken for the starch to be hydrolised.

Steps 4 to 8 are repeated for test tubes Q, R, S and T at temperatures of 28 °C, 37 °C, 45 °C and 60 °C.

The rate of enzyme reaction is calculated by using the formula

1 time (t)
1
time (t)

11 A graph showing the rate of enzyme reaction against temperature is plotted.

Discussion 1 When the starch is hydrolised, the iodine solution change from blue-black to yellow.
Discussion 1 When the starch is hydrolised, the iodine solution change from blue-black to yellow.

Discussion

1 When the starch is hydrolised, the iodine solution change from blue-black to yellow.

2 At low temperatures, the rate of enzyme reaction is low because the enzyme is not active. It takes a longer time for the starch to be hydrolysed.

3 At 37 °C, the rate of enzyme reaction is at its maximum rate because it is the optimum temperature for the enzyme's reaction.

4 At temperatures above 40 °C, the rate of enzyme reaction decreases because the enzyme is denatured. Conclusion The hypothesis is accepted. As the temperature increases, the rate of enzyme reaction increases until it reaches ,he maximum rate at 37 °C. It then decreases and stops at 60 °C.