Enzyme Investigations (Cambridge (CIE) IGCSE Biology): Revision Note

Exam code: 0610 & 0970

Lára Marie McIvor

Written by: Lára Marie McIvor

Reviewed by: Jenna Quinn

Updated on

Investigating the effect of temperature on amylase

  • Starch solution is heated to a set temperature

  • Iodine is added to wells of a spotting tile

  • Amylase is added to the starch solution and mixed well

  • Every minute, droplets of solution are added to a new well of iodine solution

  • This is continued until the iodine stops turning blue-black (this means there is no more starch left in the solution as the amylase has broken it all down)

  • Time taken for the reaction to be completed is recorded

  • Experiment is repeated at different temperatures

  • The quicker the reaction is completed, the faster the enzyme is working

Diagram of the iodine test method for investigating how temperature affects amylase activity on starch, showing results at 20°C, 40°C, and 60°C.
Investigating the effect of temperature on enzyme activity

Investigating the effect of pH on amylase

  • 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 2 cm3 of amylase in the test tube

  • Add 1 cm3 of buffer solution to the test tube using a syringe

  • Use another test tube to add 2 cm3 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 mixture on the first drop of iodine, which should turn blue-black

  • Wait another 10 seconds and place another drop of 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

Diagram showing step-by-step amylase and starch experiment using iodine on a spotting tile to measure reaction time at different pH buffer solutions (in this case pH 5, pH 7, pH 9 and pH 14.
Investigating the effect of pH on enzyme activity

Examiner Tips and Tricks

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 the top grades, can relate this to changes in the active site of the enzyme when it has denatured, or if it is a low temperature, relate it to the amount of kinetic energy the molecules have.

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Lára Marie McIvor

Author: Lára Marie McIvor

Expertise: Content Creator

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.

Jenna Quinn

Reviewer: Jenna Quinn

Expertise: Content Creator

Jenna studied at Cardiff University before training to become a science teacher at the University of Bath specialising in Biology (although she loves teaching all three sciences at GCSE level!). Teaching is her passion, and with 10 years experience teaching across a wide range of specifications – from GCSE and A Level Biology in the UK to IGCSE and IB Biology internationally – she knows what is required to pass those Biology exams.