This means the amount of substrate could have been different for each repeat, even though I used the same concentration. The diffusion limit still operates but we are now dealing with 'diffusion into a sphere of influence', rather than diffusion to the active site. I believe that the data also shows strong positive correlation, and there are few outliers, which shows that my results are accurate. This experiment was carried out by diluting the 6% H2O2 with… 3702 Words 15 Pages reactivity of the enzyme catalase on hydrogen peroxide while subject to different concentrations of an inhibitor. For example, I knew that the 100% concentration of hydrogen peroxide reaction was over because I recorded 88cm 3 at least five times. I found that the way I went about the experiment was very effective because my results matched the theory of my prediction and I got incredibly accurate results proving my experiment went positively. Catalase is an enzyme that helps to eliminate toxins such as hydrogen peroxide within an organism Nuffield Foundation.
Push the plunger on the syringe and immediately start the stopclock. Species Human 847 , Species Mouse 12359 , Species Rat 24248 , Species Goat 100860855 , Species sheep 100307035 , Species Domestic Rabbit 100340891 , Species naked mole-rat 101707368 , Species Zebrafish 30068 , Species cow 531682 , Species domestic cat 101093891 , Species dog 403474 , Species chicken 423600 , Species fruit fly 40048 Summary: This gene encodes catalase, a key antioxidant enzyme in the bodies defense against oxidative stress. Measuring of the volume of oxygen released in the burette It is a procedural error and it is subjective. This would show that the reaction is a first order reaction. I will then work out the gradient of the two concentrations and plot them on a rate of reaction graph along with the other concentrations. Perform the floating disk procedure for each concentration. Conical flask - I have chosen the conical flask instead of the boiling tube because it can stand on its own and the heat of my hands will not affect the temperature of the experiment thus will not affect the rate of reaction.
The configuration of the active site gives the specificity of enzyme. Range of time between each reading The aim of the experiment is to compare the volume of oxygen within a same time, so it was kept constant. Binding sites of relatively low affinity will increase the effective local concentration of a substrate. You might need to add water to make it less viscous and easier to use. Â· Be careful when using water baths because they can reach dangerously high temperatures Â· Use the blender appropriately as mishandling it could cause harm. The same idea applies to the substrate concentration in that the pipettes also had an apparatus error.
If there were fewer molecules of hydrogen peroxide, there would have been fewer collisions between molecules of enzyme and substrate, resulting in fewer enzyme-substrate complexes being made. For each parts A-F, you will begraphing enzyme activity on the Y-axis the dependent variable andthe independent variable parts A-F on the X-axis. Take care inserting the bung in the conical flask — it needs to be a tight fit, so push and twist the bung in with care. That is at the beginning of the reaction, the higher concentration of hydrogen peroxide, the more oxygen it evolves. That means the active site and the substrate should be exactly complementary so that the substrate can fit in perfectly. Prediction: The prediction for this investigation is that the volume of oxygen produced per unit of time increases with the increasing concentration of catalase.
The tubing was then removed from the frontal-base of the cylinder, with the ends concealed following removal. It is because every enzyme active site is working continuously. So if the apparatus is free of leaks, 22 cm 3 of water should be displaced in the measuring cylinder with 10 vol hydrogen peroxide. If so it would also be interesting to examine the peroxisomal structure described earlier to see whether its structure could enhance the effect, e. Use the pipette to measure out 3cm3 of 1% concentrated hydrogen peroxide 8.
According to the website, www. This temperature is about 91. Hydrogen peroxide is an unstable compound, which will decompose independently over time and through contact with other catalytic agents such as high temperatures and light. The methods discussed here are the slide method, tube method, and tube slant method. Given that the single Fe centre in the prosthetic heme group surely participates in the active site, it suggests that a single active site has two functionalities! Introduction In the experiment the magnesium reacts with the hydrochloric acid to create magnesium chloride and hydrogen. The syringe should be washed by distilled water before each dilution.
Using a balance weigh out 100g's of potato used as the Catalase as it contains it. All measurements that I used should be corrected to the nearest 0. In this book, the applications of a wide variety of enzymes are highlighted. Overall, there is a pattern showing a constant trend in that as the concentration decreases the rate of reaction decreases too, and that the overall volume of gas evolved also decreases. This obviously affects the amount of catalase present, which means that there could be more or fewer collisions and resulting successful collisions between enzyme and substrate molecules depending on the greater or lower mass of yeast.
The results I collected after my experiment were very accurate because it backed up my theory in my prediction and conclusion and the results were close in all three attempts with the increasing concentration resulting to a larger amount of froth. This would have affected the number of molecules of hydrogen peroxide present, which in turn would have affected the number of collisions between enzyme and substrate molecules. Sometimes, catalase test is combined with other tests such as antibiotic resistance test while it cannot identify a specific organism alone, however, catalase test remains an important tool in biological experiments. A water bath that hasbeen warmed to 37C. The reaction was fastest when the substrate concentration is 80%.
I chose Catalase for my web site domain name in part because Catalase consumes H 2O 2 in another way: The Peroxidative Reaction Catalase performs a very elegant 'reshuffling' of toxic compounds. Rotatethe reaction vessel so that the disks are on the top side seepicture above and then add 10 ml of substrate H 2O 2 solution. This extraordinary bifunctionality of Catalase led me to wonder whether there is perhaps some as yet unrecognised post translational modification or allosteric effect that specialises and fine-tunes Catalase for one or other of its two roles. If collecting the gas over water is complicated, and you have access to a 100 cm 3 gas syringe, you could collect the gas in that instead. Each piece of apparatus has an apparatus error with an upper and lower limit.
Some of the 'missing energy' is locked up in the oxidative power of H 2O 2 which is used in the peroxidative reaction described earlier. Subsequently, if there are more molecules, then there will be more collisions taking place, and therefore more reactions between enzyme and substrate molecules per second, and so oxygen is evolved more rapidly. Be aware of pressure building up if reaction vessels become blocked. Though in theory, this should be the trend, my results did not demonstrate this pattern. Catalase is extremely fast acting because it has 4 active sites per molecule which is quite unusual. It is the same in a enzyme-catalyzed reaction. If it was not immediately broken down, it will accumulate and kills cells.