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The effect of enzyme concentration on the enzyme activity

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Introduction

The effect of enzyme concentration on the enzyme activity Aim The aim of this experiment is to investigate the effect of a reduction in enzyme concentration on the rate of reaction, in this case the breakdown of protein by trypsin solution. Hypothesis I predict that as the enzyme concentration is increased the rate of reaction of the breakdown of protein by trypsin solution will increase. Scientific Reasoning Enzymes are proteins. Protein is made up of a sequence of amino acids. They are globular proteins with an active site contained in their 3 dimensional structures. The shape of the protein determines the functioning of the enzyme. The arrangement of molecules on the enzyme produces an area known as the active site within which the specific substrate(s) will "fit". Each different enzyme has its own specific shape, with a pocket, at a particular position. The pocket is known, as the active site in the location on the enzyme surface where substrates bind. Only a few amino acids, normally between 3 and 12 in the chain, of the enzyme are actually in the enzyme-controlled process. An Enzyme is also a biological catalyst found in biological organisms, which increases the speed of a chemical reaction without them undergoing any permanent chemical change. They are neither used up in the reaction, nor do they appear as reaction products. ...read more.

Middle

The activation energy is distinct from the DG, or free energy difference between the reactants and products. Apparatus * Alkali Trypsin Solution * Milk powder * Test Tubes * Stop Clock * Thermometer * 20ml, 10ml, 5ml syringes * 100ml Beaker * Distilled water * 250ml beaker Method First I will collect the equipment. As the trypsin solution that has been provided is already at 1%, I will not be able to concentrate the solution any higher, and for this reason I have decided to lower the concentration of enzyme to my desired levels. To achieve this I will have to have, depending on the concentration a ratio of distilled water and trypsin. I want to make 220ml of each concentration, e.g. to make a 0.5% concentration I will need 10mlof distilled water and 110ml of trypsin solution. The concentrations and volumes needed for the concentrations are stated in the below table. Concentration (%) Volume of Trypsin (ml) Volume of Distilled Water (ml) 1.0 20 0 0.8 16 4 0.6 12 8 0.5 10 10 0.4 8 12 0.2 4 16 0.0 0 20 Using 5ml, 10ml, 20ml syringes I will make these concentrations. When the concentrations are made, I will take at test tube and add 5ml of milk, to this I will add 5ml of trypsin solution and place it into a water bath. ...read more.

Conclusion

Also the temperature could have altered during the process of the experiment. This could have caused the enzymes to inactivate, as they would have been below the optimum temperature and this could have had caused them to stop functioning and increase the rate of the reaction. Also the pH of the enzyme could have had changed during the reaction and this could have caused the ionic bonds that hold the structure of enzyme together to split. This would affect the charges on the amino acids within the active site and the enzyme will not be able to form an enzyme substrate complex. Trypsin works best at a pH of 8 and if this alters it will cause the ionic bonds of trypsin enzyme to split, and lead to a change in the reaction. To improve the experiment I could use more accurate equipment and take more readings to see if the errors are a one off situation. It also would help the validity of results if I varied the substrate concentration to see the maximum activity. Also I could record by using the data logger or the colorimeter, which is a more reliable and accurate way of recording colour-changing results. Also I could use magnetic stirs and automated pipettes and burettes instead of the syringes and this would have also helped improve the validity of the results. Overall I am happy with the way I have gone about the experiment with the given equipment. Ibrar Razaq 12PWR - 1 - ...read more.

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