Investigating the effects of Temperature on Amylase.

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Investigating the effects of Temperature on Amylase

Aim:

The aim of this investigation was to record and analyse how changing the temperature of amylase before exposing it to starch solution affects the time taken for the amylase to take affect on the starch and break it down to produce Glucose.

        By increasing the  temperature the energy supplied to the enzyme will increase, thereby increasing the number of vibrations occurring within the amylase, this increase in movement increases the possibility of the amylase enzyme binding with a starch substrate and breaking it down into glucose .   The rate at which the enzyme breaks down the substrate will continue to increase until it reaches the optimum temperature; this is the point at which the enzyme is most efficient suboptimum temperatures will continue to break down the substrate but less effectively.  Increasing the temperature slightly above the optimum will decrease the efficiency of the process and will eventually denature the enzyme rendering it useless.  This occurs because the active site of the enzyme where the substrate binds with it and is broken down into its product changes shape.  This new shape does not fit that of the substrate therefore the enzyme activity is lost.  The optimum temperature of enzymes is relative to their natural environment, as is the case with other variables such as PH.  Enzymes within the human body will work best at roughly 370C as this is the internal body temperature.

        

Preliminary work was carried out to determine what concentrations of amylase and starch solution to use.

Preliminary Work:

In this investigation it was decided that for the preliminary work different concentrations of amylase and starch solution would be tried and tested and the concentrations that worked the best would be picked.  During the work it became apparent that this would not be the case as the initial concentrations of 5% starch solution and 3% amylase were too concentrated and positive tests for glucose were being recorded as early as 20seconds into the test.  From this the experiment was refined and smaller concentrations of amylase and starch solution were sought. Upon retesting it was decided that the concentrations of 2.5%starch solution and 1% amylase would be used as upon testing at room temperature a positive test for starch was recorded after 3 minutes this concentration.  Below is a results table showing how through trial and error the concentrations were chose for the investigation:

The final concentrations listed were used as room temperature would be the test that takes the longest to produce glucose and 3 minutes as the longest waiting time is fitting considering the number of tests that must be done and the time constraints.

        

Variables:

The major variable is obviously temperature which was changed in a controlled fashion and raised by 10 degrees in each test.  The concentration was kept the same throughout the experiment for both the amylase and the starch to ensure a fair test, the concentrations were chosen through the preliminary work. No specific measures were taken to keep other potential factors from changing the natural course of the experiment as the other major variables such as PH, Oxygen and Moisture will in no way be significant enough to alter the natural course of the experiment.  This assumption was made on the basis of the preliminary work where the tests were carried out under the same conditions that the real experiment would be done and the results that were collected were feasible thus showing that no extra measures need to be taken to keep the listed variables constant.  General precautions such as using similar equipment and material each time were observed to obtain as accurate results as possible.

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Prediction:

I know that all enzymes have different optimum temperatures at which they work best and after that they continue to work but soon become denatured.  This occurs when the bonds within the amylase are shaken apart collapsing the 3D structure of the protein destabilises.   Also enzymes have a structure which they hold best at their optimum temperature so at the optimum which I expect that the enzyme will work better as it holds this shape.  Enzymes that work better at higher temperatures tend to have a more rigid structure.  I know that most enzymes work best ...

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