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Effects of immobilisation on the rate of reaction of an enzyme at different temperatures and pH.

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Rationale Biology - Effects of immobilisation on the rate of reaction of an enzyme at different temperatures and pH. The aim of the experiment is to study closely and document the effects of immobilisation on the rate of reaction on an enzyme at different temp/pH. To do this I plan to do a few preliminary experiments that will show the effect of temp and pH on enzymes. I will also do a preliminary to show effect of immobilisation on enzymes. I'm carrying out this research so I can show how immobilisation affects enzymes in different situations. The questions I am trying to answer are * Does immobilisation affect the rate of reaction of an enzyme and does changing the temp or pH affect these results. Background Information Enzymes were discovered by the German chemist Eduard Buchner towards the end of the nineteenth century. He had been trying to extract from yeast a fluid of medicinal use, but the extract kept going bad. To prevent this he tried adding sugar to the yeast and found that the sugar was converted into alcohol: in other words it fermented. Buchner showed that the living cells were not responsible for fermentation but that the fluid extracted from them was. The word enzyme was coined, for the active ingredients in the juice that promotes fermentation. Enzyme literally means "in yeast", but now it is used as the collective noun for many hundreds of compounds that have been extracted from cells and shown to have a catalytic action on specific chemical reactions. ...read more.


An example would be the micro organisms which are capable of living in hot springs at temperatures of 1000c and above. If we go back to the diagram again we can see that at a low temperature such as 100c, nearly nothing is happening, this is because molecules are moving relatively slowly. Substrate molecules will often not collide with the active site, and so bonding between substrate and enzymes doesn't often occur. As temperature rises, the enzyme and the substrate molecules move faster. Collisions happen more frequently, so the rate of reaction increases, causing more products to be made. Also when they do collide, they do so with more energy, meaning there is an increased chance of bonds being broken and positive reactions occurring. As temperature continues to increase, the speed of enzyme and substrate molecules continue to increase. However as mentioned earlier, above a certain temperature the structure of the enzyme molecule vibrates so energetically that some of the bonds holding the enzyme in its precise 3D shape begin to break down. This is the basis of denaturing, which is often irreversible once it has begun. At first the substrate molecules will fit less well into the active site of the enzyme so the rate of reaction slows down. Eventually the substrate no longer fits at all, or it can no longer be held in the right place, long enough for the reaction to occur. The temperature at which an enzyme works best is called its OPTIMUM TEMPRETURE. ...read more.


It is desired to avoid reaction with the essential binding site group of the enzyme. Alternatively, an active site can be protected during attachment as long as the protective groups can be removed later on without loss of enzyme activity. In some cases, this protective function can be fulfilled by a substrate or a competitive inhibitor of the enzyme. The surface on which the enzyme is immobilized is responsible for retaining the structure in the enzyme through hydrogen bonding or the formation of electron transition complexes. These links will prevent vibration of the enzyme and thus increase thermal stability. The micro environment of surface and enzyme has a charged nature that can cause a shift in the optimum pH of the enzyme of up to 2 pH units. This may be accompanied by a general broadening of the pH region in which the enzyme can work effectively, allowing enzymes that normally do not have similar pH regions to work together. * Carrier-Binding : the binding of enzymes to water-insoluble carriers * Cross-Linking: intermolecular cross-linking of enzymes by bi-functional or multi-functional reagents. * Entrapping : incorporating enzymes into the lattices of a semi-permeable gel or enclosing the enzymes in a semi-permeable polymer membrane Conclusion I will use this information to try and conclude whether immobilization affects rate of reaction of enzymes at different temperatures and pH's. I will first carry out some preliminary experiments to determine the effect of pH and temperature on enzymes, before studying effect f immobilization.This will give me a comparative in order to determine the true meaning of my results. ...read more.

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