How does varying bacterial amylase effect inhibition by copper?
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INVESTIGATION How does varying bacterial amylase effect inhibition by copper? Amylases are enzymes, which hydrolyse starch into maltose. There are two main types of amylase: - 1) Alpha amylase, which degrade starch molecules into fragments 10 glucose residues long; 2) And there are Beta amylases, which break down into maltose made of two glucose molecules. Both are working by hydrolysis adding one molecule of water across the glycosidic link. Hypothesis I think that the higher the concentration of bacterial amylase, the faster the rate of reaction. This is because the higher the concentration the faster the rate of reaction. Increased concentration results in an increased possibility that collisions between molecules with the required activation energy will occur. Copper is non-competitive inhibitor. In non-competitive inhibition, the inhibitor may form a complex with the enzyme itself, with the enzyme/substrate complex or with the prosthetic group. The inhibitor is not competing for the active site but joins to the enzyme molecules elsewhere. Bacteria use amylase to feed and therefore provide energy to the cell. Bacteria are prokaryotes. Prokaryotes are made up of prokaryote calls. They do not have a membrane-bound nucleus. The genetic material is in a single strand coiled in the centre to form a nucleiod. They are also single celled organisms. We cannot live without bacteria, yet bacteria causes may diseases, which kill many people, for example TB, typhoid, cholera, and many more.
Therefore I am doing another preliminary test with lower concentration of iodine solution. I will use 2ml of 0.25% bacterial amylase, 2ml of 2% starch and 0.1% iodine. I will not add copper sulphate solution and the temperature will be at room temperature. % Of iodine solution Start time in secs Finish time in secs 0.1% 0 347 This shows that the experiment works but the experiment takes too long so I will repeat the experiment at different temperatures. The temperature range I will use will be from room temperature, 20 to 80°C. To do this I will need to set up water baths at different temperature. Temperature in °C Start time in secs Finish time in secs Time taken in secs 20 0 353 353 30 9 281 272 40 15 268 253 50 22 260 238 60 30 317 287 70 37 477 440 80 45 No reaction N/A This shows the optimum temperature for the reaction. The optimum temperature is in between 40 and 50°C so I will use 45°C. Therefore I will do the preliminary experiment I planned to do to find out the concentration of copper sulphate solution to use. % Of copper sulphate solution Start time in secs Finish time in secs Time taken 1% 0 579 579 2% 8 1063 1055 3% 15 1457 1442 4% 20 N/A N/A 5% 26 N/A N/A No copper sulphate added 32 269 237 This shows the results for the preliminary experiment to find out the concentration of copper sulphate solution to use.
This changes the shape of the enzyme and as a result, reduces the rate of the reaction. As the concentration is increased, there are more enzymes in the solution than there are inhibitors. This means that more of enzymes are involved in the reaction, therefore the rate of reaction increases. This indicates that my hypothesis 'I think that the higher the concentration of bacterial amylase, the faster the rate of reaction.' was correct. EVALUATION The main problem of my experiment was to determine the end point of the experiment was. The colour change from the blue/black to colourless was difficult to determine. Attaining the correct concentration of bacterial amylase was the difficulty I encountered whilst conducting the experiment. In order to attain the correct concentration, I had to dilute the bacteria amylase using distilled water. Overall, I believe that the experiment was accurate and that I acquired accurate results. I repeated the experiment three times in order to achieve accurate results and also rule out the possibility of 'fluke' or chance results. The experiment produced no anomalies. The results provided evidence that supported my hypothesis. I was able to accept my hypothesis to be correct and conclude that the rate of reaction increases with increasing concentration of bacterial amylase. To broaden my investigation for example, I could: a) Change the temperature; b) Change the ph; c) Alter the concentration of substrate/ inhibitor; d) Use different types of enzymes. There are many variables that could be altered in order to broaden my investigation and take it into another level.
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