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Enzyme required practical

The document outlines a practical investigation to study the effect of pH on the reaction rate of the enzyme amylase in breaking down starch. It details the method of using iodine as an indicator and suggests adjustments for temperature control and the use of a colorimeter for accurate measurement. Additionally, it emphasizes the importance of understanding enzyme behavior for exam preparation.

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1 views

Enzyme required practical

The document outlines a practical investigation to study the effect of pH on the reaction rate of the enzyme amylase in breaking down starch. It details the method of using iodine as an indicator and suggests adjustments for temperature control and the use of a colorimeter for accurate measurement. Additionally, it emphasizes the importance of understanding enzyme behavior for exam preparation.

Uploaded by

Kaizer Kaizer
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
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Enzyme required practical

 Aim: To investigate the effect of pH on the rate of reaction of


amylase

 You will:

o Use the enzyme amylase to breakdown starch at a range of


pH values, using iodine solution as an indicator for the
reaction occurring

o 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

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 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 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 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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