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

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.

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

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  1. 4.3 (a) To investigate the effect of pH on amylase activity

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

  3. Step 1 Set up a water bath at 37 °C.

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

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

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

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

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

  9. Graph your results

  10. Step 7 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.

  11. Step 8 A brick red colour indicates a reducing sugar is present.

  12. Step 9 Repeat the whole experiment with boiled amylase as a control.

  13. Expected result The enzyme works best at a pH of 7. The blue / black colour faded and Benedicts resulted in a positive test.

  14. Results

  15. END

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