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Investigating the Effect that Copper Sulphate has on the Action of the Enzyme Catalase

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Introduction

Investigating the Effect that Copper Sulphate has on the Action of the Enzyme Catalase Elizabeth Dodwell PLANNING Introduction: Catalase is a red, crystalline enzyme that consists of a protein complex with haematin groups and catalyses the decomposition of hydrogen peroxide into water and oxygen: 2H2O2 --> 2H2O + O2 Hydrogen peroxide is a powerful oxidising substance. It is formed inside cell organelles called the peroxisomes, during the breakdown of purines. Peroxisomes help to oxidise many substances that might be toxic to the cell. Hydrogen peroxide and catalase, an important oxidase enzyme, act together to oxidise toxins. The peroxisomes in liver cells use the catalase-hydrogen peroxide system to detoxify alcohol that a person drinks. Hydrogen peroxide is also produced in neutrophils and macrophages, two types of cells that help destroy infectious agents such as bacteria. Hydrogen peroxide is lethal to most bacteria even in very small amounts. The hydrogen peroxide-catalase oxidising mechanism is also used to break down fatty acids into acetyl-CoA that is then used for energy by the cell. Peroxisomes are small, membrane-bounded vesicles that provide a contained environment for reactions where dangerously reactive hydrogen peroxide is generated and degraded. Membranes form numerous other small vesicles involved in the transport of materials between one organelle and another. In a typical animal cell the membrane-bounded organelles may occupy up to half the total cell volume. Free radicals are produced in most cells of the body as a by-product of metabolism, although some cell types manufacture larger quantities for specific purposes (for example, by macrophages during phagocytosis). The most important free radicals found in aerobic cells, such as those in humans, are oxygen, superoxide, hydroxyl radical, hydrogen peroxide and the transition metals. When free radicals form within cells they can oxidise biomolecules (molecules used inside cells, especially lipids) and thus cause cell death and injury. However, the human body has developed various mechanisms in order to protect itself from the damaging effects of free radicals. ...read more.

Middle

Referring to each experiment by its allocated letter will simplify the format of my results (i.e. A2 indicates the second repeat for a concentration of 0M, F3 indicates the third repeat for a concentration of 2M, etc.). * 6 pieces of liver with masses of 1.5g are weighed out making sure that the surface areas are as similar as possible. * Each piece of liver is put into each CuSO4 solution and left for 10 minutes. * As apparatus are set up as shown below: * A piece of liver is put in the conical flask, then 2cm� of H2O2 is added and the bung replaced. * The volume of O2 released is measured every 30 seconds for 2 minutes. The Results from My Experiment: Time / s Volume of O2 Collected / cm� A1 A2 A3 B1 B2 B3 C1 C2 C3 D1 D2 D3 E1 E2 E3 F1 F2 F3 30 20 31 12 14 8 15 5 4 12 6 4 5 3 0 0 0 0 0 60 30 36 22 27 13 27 8 10 18 12 9 8 6 0 2 0 0 0 90 38 38 29 37 20 35 13 15 25 19 16 14 9 4 5 0 2 4 120 39 38 29 37 28 38 19 19 29 21 19 15 13 9 10 0 5 7 Average Values Obtained from my Results: Time / s Volume of O2 Collected / cm� A B C D E F 30 21 12 7 5 1 0 60 29 22 12 10 3 0 90 35 31 18 16 6 2 120 35 34 22 18 11 4 I also obtained results from fellow students: #1 Time / s Volume of O2 Collected / cm� A B C D E F 30 16 8 4 2 1 0 60 24 15 8 4 3 1 90 32 25 12 8 4 2 120 38 35 17 11 5 4 #2 Time / s Volume of ...read more.

Conclusion

This liver could not have been at exactly the same temperature as the liver I was using before. I should have ensured that there was a plentiful supply of liver that had been left to acclimatise for a sufficient amount of time before I used it. Also, as I had to keep using the same glassware, a significant temperature change could have occurred through rinsing in cold water. Had there been an adequate supply of glassware, this problem would have been eradicated. Was I to repeat this experiment I would make significant changes as described above in order to obtain a higher degree of precision in my results. However, despite there being considerable room for improvement in my method, I am confident that the results I obtained were sufficient in order to prove my hypothesis. The graphs formulated from these results showed trends that I expected to see and the trends observed over the whole range of graphs were consistent. They clearly show the effect that copper sulphate has on the action of the enzyme catalase despite errors being made. In order to find out exactly what type of inhibitory effect copper sulphate has on the action of the enzyme catalase, further experimental procedures would have to be followed. If the rate of reaction increased when more substrate was added, it would be a competitive or non-competitive reversible inhibitor; if the rate of reaction did not increase when more substrate was added and merely continued to slow down, it would be a non-competitive irreversible inhibitor. Further experiments beyond my scientific knowledge would be required to differentiate between a non-competitive and competitive reversible inhibitor Had all the variables been precise throughout the experiment I think that everyone's graphs would have shown identical curves with the same values for each reading throughout a range of repeats. The errors made were due to several factors: insufficient planning prior to performing the experiment, personal error, inappropriate equipment, insufficient repeats etc. ...read more.

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