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The Effect of the Concentration of an Inhibitor on The time taken for the Enzyme to Fully Breakdown the Substrate

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Introduction

The Effect of the Concentration of an Inhibitor on The time taken for the Enzyme to Fully Breakdown the Substrate Aim For this investigation I am going to investigate the effect of the concentration of an inhibitor, on the time taken for the enzyme to fully breakdown the substrate. Introduction I am going to complete my aim by planning an experiment, carrying out the experiment, recording any relevant results and plotting graphs from which I will be able, hopefully to gain a strong conclusion. I will finally evaluate the whole investigation. I will be using human amylase to breakdown the substrate starch. The reason this enzyme has to be used is because each enzyme is designed specifically to break down only one substrate as each enzyme is made of a protein that causes it to be a specific shape, in this case the enzyme Amylase can only break down starch. Also the inhibitor I will be using is copper sulphate, a heavy metal ion. Background Reading Amylase, like other enzymes, works as a catalyst, i.e. it is unchanged by the reaction, but makes the reaction easier by reducing the energy required for it to happen. Catalysts speed up the reaction. The theory behind the working is called the "lock and key" theory: the enzyme is shaped so that the products fit into them, react and are released. Amylase digests starch by catalysing hydrolysis, which is splitting by the addition of a water molecule. ...read more.

Middle

Where the inhibitor is limiting the rate of reaction. I varied the inhibitor concentration by altering the mass of copper sulphate in 100ml. I decided to use a permanent inhibitor (heavy metal ion) so that the temporary effect of reversible inhibitor does not affect my results. Apparatus Needed * Test Tube Rack * 4 x 5ml syringes * 1 x10ml syringe * 12 x 10 mm diameter Test Tubes * A stop clock * A 250ml Beaker * Thermometer * Ph 7 Buffer * Spotting Tile * Bunsen burner (with Tripod and gauze) * Iodine (with pipette) * Stirring Rod * Kitchen towel * A scale * 8 x 15 ml beakers * Tongs Reason for Equipment * 15 x 10mm diameter test tubes will be used for mixing the amylase with the starch also copper sulphate and pH buffer in the water bath. * The test tubes have to be clean to prevent any unwanted contaminants getting into the experiment. 4x 5ml and 1 x 10 ml (for water) syringe used for very accurate measuring of the amylase, starch solution, water, and copper sulphate. This will be vital for getting the correct volume of the 3 substances into the test tubes and the correct amount of water for dilution. * Thermometer, the most accurate way of measuring the temperature of the water bath. Therefore the temperature of the enzyme and the substrate as well. ...read more.

Conclusion

This is because overall trends between the inhibitor concentration and rate of reaction can be portrayed more effectively and become more obvious Limitations and Precautions I monitored the temperature using a thermometer to ensure that it remained constant and not disrupt the results of the experiment by affecting the activity of the amylase A pH buffer was used to maintain a consistent pH level in the test tubes. This way there was no variation in pH that might have resulted in an increase or decrease in the rate of reaction. A major limitation of this investigation was the time. It meant that only 6 different inhibition concentrations could be measured at intervals 0.01 moles. This means that only very general, overall trends can be identified across the results. Patterns between these values can only be approximated and are not necessarily accurate. Safety Laboratory coats were worn during the investigation to prevent chemicals from spoiling clothes. Safety aspects Goggles must be worn and especially while using the iodine as it can be irritable to the eyes. Stand up while using the Bunsen burner so you can move away quickly should the water bath fall. Any long hair tied back to keep out of way of flames. Results Concentration Experiment 1(time) Experiment 2 (time) 0.01 300.00 300.00 0.02 480.00 480.00 0.03 540.00 540.00 0.04 1020.00 1020.00 0.05 1260.00 1260.00 0.06 2100.00 2100.00 Concentration Average (time) 0.01 300.00 0.02 450.00 0.03 600.00 0.04 960.00 0.05 1290.00 0.06 2040.00 Concentration Average (rate of reaction) 0.01 3.33 0.02 2.22 0.03 1.67 0.04 1.04 0.05 0.78 0.06 0.49 Conclusion ...read more.

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