Required Practical: Enzymes (AQA GCSE Biology: Combined Science)

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Enzyme required practical

  • Aim: To investigate the effect of pH on the rate of reaction of amylase
  • You will:
    • Use the enzyme amylase to breakdown starch at a range of pH values, using iodine solution as an indicator for the reaction occurring
    • Use a continuous sampling technique to monitor the progress of the reaction
  • Amylase is an enzyme that digests starch (a polysaccharide of glucose) into maltose (a disaccharide of glucose)
  • Starch can be tested for easily using iodine solution

 

using-iodine-to-test-for-starch-igcse-and-gcse-biology-revision-notes

Iodine can be used qualitatively to indicate the presence or absence of starch from a sample

Method

  • Place single drops of iodine solution in rows on the tile
  • Label a test tube with the pH to be tested
  • Use the syringe to place 2cm3 of amylase in the test tube
  • Add 1cm3 of pH buffer solution to the test tube using a syringe
  • Use another test tube to add 2cm3 of starch solution to the amylase and buffer solution, start the stopwatch whilst mixing using a pipette
  • After 10 seconds, use a pipette to place one drop of the mixture on the first drop of iodine, which should turn blue-black
  • Wait another 10 seconds and place another drop of the mixture on the second drop of iodine
  • Repeat every 10 seconds until iodine solution remains orange-brown
  • Repeat experiment at different pH values – the less time the iodine solution takes to remain orange-brown, the quicker all the starch has been digested and so the better the enzyme works at that pH

investigating-effect-ph-on-enzyme-activity-igcse-and-gcse-chemistry-revision-notes

Investigating the effect of pH on enzyme activity

Adjustments to the method

  • The above method can be adapted to control temperature by using a water bath at 35°C
  • All solutions that need to be used (starch, amylase, pH buffers) should be placed in a water bath and allowed to reach the temperature (using a thermometer to check) before being used
  • A colorimeter can be used to measure the progress of the reaction more accurately; with a solution containing starch being darker and glucose light (as a result of the colour-change of iodine) – this will affect the absorbance or transmission of light in a colorimeter

Examiner Tip

Describing and explaining experimental results for enzyme experiments is a common type of exam question so make sure you understand what is happening and, for a 7, 8 or 9, can relate this to changes in the active site of the enzyme when it has denatured.

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Lára

Author: Lára

Expertise: Biology Lead

Lára graduated from Oxford University in Biological Sciences and has now been a science tutor working in the UK for several years. Lára has a particular interest in the area of infectious disease and epidemiology, and enjoys creating original educational materials that develop confidence and facilitate learning.