4.3 (a) To investigate the effect of pH on amylase activity

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4.3 (a) To investigate the effect of pH on amylase activity

Before you start get or make a copy of this Table of results

Step 1 Set up a water bath at 37 °C.

Step 2 Place 5 ml starch solution and 3 drops of iodine solution into 3 boiling tubes.

Step 3 Put 2 ml amylase solution and 5 ml buffer (4, 7, 9) into three other tubes.

Place all tubes into a water bath at 37 °C and leave for 5 minutes. Step 4 Place all tubes into a water bath at 37 °C and leave for 5 minutes.

Step 5 Pour one of the amylase tubes into one of the starch tubes. Gently shake. Repeat for each pH.

Step 6 Replace the tubes in the water bath and measure the time taken for the blue/black colour to disappear.

Graph your results

Prepare as for steps 1 and 2 previously Test 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 a pH of 7. Expected result The enzyme works best at a pH of 7. The blue / black colour faded and Benedicts resulted in a positive test.

Results

END