Investigate the effect of temperature on enzyme (amylase) activity.

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Investigating Enzymes

Aim: - 

To investigate the effect of temperature on enzyme (amylase) activity.

Introduction & Background Information

As mentioned above, the aim of this experiment is to determine whether temperature has an effect on enzyme activity or not. To test this starch will be hydrolysed in to maltose with the enzyme Amylase. Fearons reagent will then be added to determine the colour and thus the amount of maltose present. This reagent will indicate the relative amounts of maltose produced at each temperature by colour comparison. The colour intensity’s will then be examined by a calorimeter to provide us with the amount of absorbance.

Starch                         Maltose

                                Add Fearons Agent

                                        (Turns Red)

                                Intensity examined by a Calorimeter

The following shows how enzymes work : -

Hypothesis: - Enzymes work best at an optimum temperature, which is 37.5oc in the human body. An increase in temperature supplies more kinetic energy to the molecules. The numbers of collisions between enzyme and substrate will thus increase so the rate of reaction will increase too. Above the optimum temperature, and the enzymes are denatured. Bonds holding the structure together will be broken and the active site loses its shape and will no longer work. This hypothesis will now be tested in this experiment.

Safety precautions 

Safety goggles and a lab coat would be worn at all times. All parts of the experiment would be undertaken with care, to ensure that there was no spillage. Any spillages of NaOH or starch would be washed thoroughly with soap and water. If any starch solution, NaOH or amylase was accidentally ingested, medical attention would be sought immediately. Extra care should also be taken when handling the Bunsen burner, i.e. long hair should be tied back.

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Apparatus

0.5 % starch buffered to pH 6.7

1% NaCl solution

Fearons reagent

20% NaOH solution

pH 6.7 buffer

Amylase solution

Distilled water

Tongs

Boiling tubes

Boiling tube rack

100 cm3 measuring cylinder

1, 5 and 10 cm3 pipettes and safety pumps

Gloves

Goggles

Cuvettes

Lab Coat

Stop clock

Colorimeter

Ice Bath (0 oc), 21oc , 30 oc, 40 oc, 50 oc, 60 oc, 70 oc  water baths

Bunsen burner

Tripod

Gauze

Large Beaker (500ml)

Heatproof Mat

Method: -

  1. Put lab coat, goggles and gloves on.

  1. Prepare 7 tubes of starch (labelled S1 to S7) each containing the ...

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