4ml 4ml 4ml 4ml 4ml Total Volume 5ml 5ml 5ml 5ml 5ml 5ml After setting those

# 4ml 4ml 4ml 4ml 4ml total volume 5ml 5ml 5ml 5ml 5ml

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4ml 4ml 4ml 4ml 4ml Total Volume 5ml 5ml 5ml 5ml 5ml 5ml After setting those test tubes to the side, we prepared a water bath at -4 . We took the substrate test tube for test one and the enzyme test tube for test one (as described in Table 3 and Table 4) and incubated them separately in the -4 water bath for five minutes. We used a timer to ensure we had the most accurate data as possible. After the five minutes were over, we mixed the two solutions together in a cuvette and placed it immediately in the cuvette chamber. We recorded the absorbance every thirty seconds for 180 seconds, then removed the cuvette from the chamber, We repeated this process with the remaining corresponding tubes one at a time. Those water baths were 22 , 35 , 50 , 65 , and 80 . Once we finished recording the data, we
7 discarded the solutions and cleaned up the experiment. Results/Data The data collected from the experiment was put into graphs and tables to clearly show trends and connections between temperature and pH conditions on enzyme reaction speeds. Table 5: pH Data Time (in seconds) Test 1 (pH 4) Test 2 (pH 5) Test 3 (pH 6) Test 4 (pH 7) Test 5 (pH 8) Test 6 (pH 9) 30 0.013 0.191 0.159 0 0.06 0.011 60 0.055 0.359 0.354 0 0.132 0.024 90 0.105 0.534 0.54 0 0.207 0.035 120 0.153 0.739 0.755 0 0.264 0.045 150 0.202 0.979 1.03 -0.001 0.336 0.055 180 0.247 1.344 1.505 0 0.405 0.063 Graph 1: Effect of pH on Enzyme Reactions Table 5 and Graph 1 demonstrate the results and data collected during the pH portion of
8 the experiment. Generally, the absorbance of each solution increased every 30 seconds, regardless of the buffer used. Test 3 with a pH of 6 had the highest absorbance rate at 1.505. Test 4 with a pH of 7 had the lowest absorbance and dipped to -0.001 at 150 seconds. Test 1, Test 5, and Test 6 all increased at a steady speed, forming a linear pattern on Graph 1. Test 4 did not increase at all, forming a straight horizontal line with a slight dip at 150 seconds. The absorbance of Test 2 and Test 3 exponentially increased over time and started with a higher absorbance than the others at 0.191 and 0.159, respectively. These two tests absorbed the most amount of light much quicker than the other tests. Test 4 had a negative absorbance and, therefore, absorbed no light. Table 6: Effect of pH on the Reaction Rate Test 1 Test 2 Test 3 Test 4 Test 5 Test 6 Rate of Reaction 0.0014 0.0075 0.0084 -0.0000056 0.0023 0.0004 Table 6 illustrates the rate of the enzyme reactions for each test. The reaction rate is the amount of change per second. Clearly, Test 2 and Test 3 had very close reaction rates and changed more per second than any other test. Test 4 had the least amount of change and was the only test with a negative reaction rate. This data correlates with Table 5 and Graph 1. Table 7: Temperature Data Time (in seconds) Test 1 (4 ) Test 2 (22 ) Test 3 (35 ) Test 4 (50 ) Test 5 (65 ) Test 6 (80 ) 30 0.087 0.11 0.145 0.243 0.232 0.02 60 0.157 0.188 0.245 0.384 0.339 0.031 90 0.219 0.26 0.325 0.498 0.43 0.045 120 0.274 0.322 0.39 0.59 0.506 0.058 150 0.325 0.378 0.452 0.666 0.568 0.069
9 180 0.375 0.428 0.484 0.724 0.616 0.081 Graph 2: Effect of Temperature on Enzyme Reactions Table 7 and Graph 2 expresses the results and data from the temperature portion of the experiment. Overall, the absorbance of a solution increased over time. As the temperature of a

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• Fall '14
• Enzyme, pH, Chemical reaction, pH Substrate Test Tubes

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