Comparing Free to Immobilised Amylase Enzyme in Its Catalysis Rate

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Comparing Free to Immobilised Amylase Enzyme in Its Catalysis Rate


First of all, the Immobilised enzymes need to be made. The method used to create these immobilised enzymes would be Micro encapsulation. This means that the enzyme used, in this experiment being Amylase, is encapsulated inside Sodium alginate. The enzyme was believed to act quickly, so the enzyme would have to be slightly diluted in order to get a good range of results. !0cm³ of Amylase will be added to 20cm³ of Sodium alginate, and then mixed. Using a pipette, the Sodium Alginate - Amylase mix is dropped into Calcium Chloride, and this will form the beads of Immobilised enzymes needed for the experiment. As soon as
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The product made from adding Amylase and Starch together is Maltose, which is a Disaccharide, so this is in essence, a reducing sugar test. Previous data from other experiments will be used to compare methods in order to maximize the experiments efficiency.

In this experiment, there are many variables to consider. Temperature must be kept at a constant, and at optimum for the Amylase, being at 35°c. Thermometers will be used to monitor the temperature and a water bath will be used to heat the experiment. P.H. must also be kept at optimum and this will be achieved by adding Buffer solution to the enzyme and substrate mix. The Buffer solution will keep the mixture at p.H. 7.2, which again, is optimum for the Amylase. The overall volume of enzyme and substrate must be kept the same, and hence concentrations must be kept the same too. The overall volume will be 20cm³ and the concentrations will be kept the same. The independent variable chosen is time. There will be different time periods that the enzyme and substrate are kept together for both immobilised and free enzymes. There will be 30-second intervals between each removal of the samples. Time is the independent variable because the catalysis rate is being measured of immobilised enzymes and free enzymes, so time is the obvious choice. The rate of

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