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4.4 (a) To investigate the effect of temperature on amylase activity
Before you start get or make a copy of this Table of resultsTemperature (°C) Time taken for solution to turn colourless Test for sugar
Step 1 Set up five water baths at different temperatures, e.g. 0°C, 20°C, 35°C, 55°C, 80°C.
Step 2 Into each of five boiling tubes put 5 ml starch solution and 3 drops iodine solution.
Into each of five more boiling tubes put 2 ml amylase solution.Step 3 Into each of five more boiling tubes put 2 ml amylase solution.
Step 4 Place a pair of boiling tubes into each water bath and leave for 5 minutes.
Step 5 Pour the amylase into the tube of starch solution and gently shake. Repeat for each temperature.
Step 6 Replace the tubes in the water bath and measure the time taken for the blue/black colour to disappear.
Prepare as for steps 1 and 2 previouslyTest for sugar: add 1ml Benedict’s solution to each boiling tube and heat for 5 minutes. Prepare as for steps 1 and 2 previously
A brick red colour indicates a reducing sugar is present.Step 8 A brick red colour indicates a reducing sugar is present.
Repeat the whole experiment with boiled amylase as a control.Step 9 Repeat the whole experiment with boiled amylase as a control.
The enzyme works best at 37OC.Expected result The enzyme works best at 37OC. The blue / black colour faded and Benedict’s resulted in a positive test.
Table of Results Temperature (°C)Time taken for solution to turn colourless Test for sugar
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4.3 (a) To investigate the effect of pH on amylase activity
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Test for simple (reducing) sugars Benedict’s is blue Put test substance in a test tube with distilled water and a few drops of Benedict’s Heat.
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