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PRACTICAL- ENZVlViE CONCENTRA'l'ION AND RATE OF


REACTION 0 (j

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AIM: To investigate the effect or a reduction in enzyme concentration on the rate of reaction by
measuring the initial rate of reaction,
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BACKGROUND: the viscosity of afixed volume!of gelatine cail be determined by the time
taken for the jelly to drain through a funnel. if the gelatine is treated with
the enzyme pepsin and the viscosity determined this will give a rneasure
C?fthe breakdown (l the protein by fhe enzyme.
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HYPOTHESIS: (what do you think will be the relationship ~etween protease.concentration


and the rate of breakdown of protein in gelatine? Use scientific ideas to
support your prediction).
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APPARATUS LIST: /
. 75g of2% gelatine solution (fully set) pH 2.8 bufTer pepsin solu~ion (5%)
. Funnel 2 pipcttcs
1
stopclock test tubes
41 Test tube rack spatula. 2 X 250cm3 beakers
41
Glassware for diluting enzymcs eye protection

METHOD

1. Suspend a funnel over an empty beaker.

2. Pipette 3cm3 of pH 2.8 bufTcr into a test tubc and place in a test tube rack.

3. Pipette 7cm3 of5% pepsin solution into the test tube.


4. Stm1thc stopclock as the mixture in the test tube is poured into a beaker containing 75g
of gelatine. Stir the contcnts gently for 15 seconds using a spatula then leave for 5
minutes to stand.

5. Pour the mixture into the funnel using the spatula to empty the beaker.

6. Restart the stopclock as the first drip Icavcs the funnel. Rccord the time taken for all
the gelatine to pass through the funnel.

7. Repeat the test twice more and calculate an average time.

8. Repeat the experiment with a range of different enzyine concentrations by diluting the
5% pepsin solution (see below).
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Enzyme conc % .0 1 2 I 3 4 5
V01 of 5% pepsin sol/cm-:1 0 1 2 J 4 5
Vol of distilled water/cmJ 5 4 J 2 1 0
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RISK ASSESSMENT (identify any risks and list safety precautions) /:


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RELIABILITY AND ACCURACY OF RESULTS (how will you ensure that the results are
accurate and reliable? How will you ~nsure that mcasurements arc precise, accuraFe and
repeatable? Include replicates and lIse of a control).
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IDENTIFICATION OF DEPENDENT AND INDEPENDENT VARIABLES

1. DEPENDENT VARIABLE =:= C0~1\ C ' c1 ; Q.Y"1-~tJ 0u+ . ;


COv1C
2. INDEPEJ\TDENTVARIABLE =:= p~'7'1,/ pv-e-ss,C\...v-I2-./~p~ S\A..bjt.
3. OTHER VARIABLES =:= 8\~,