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Aayush Desai

7-11-10

The Effect of Enzyme (Catalase) Concentration on the Rate of Reaction (when Hydrogen Peroxide reacts with 1 cm3 fresh liver and the amount of foam produced) Design Aim: To find how the different substrate concentrations of hydrogen peroxide (6%, 4%, 2%, 1%, 0.5% and 0.0%) influences the rate of the reaction after putting 1 cm3 of fresh liver into the measuring cylinder and then measuring the amount of foam that is produced when the reaction occurs in cm3 using the scale on the measuring cylinder. Hypothesis: Null Hypothesis: If we put 1 cm3 of fresh liver into the measuring cylinder with different concentrations of hydrogen peroxide (6%, 4%, 2%, 1%, 0.5% and 0.0%), then the amount of foam formed in cm3 will be the same in all cylinders after 30 seconds; because there is isnt any significant difference between the different hydrogen peroxide concentration solutions. Alternate Hypothesis: If we put 1 cm3 of fresh liver into the measuring cylinder with different concentrations of hydrogen peroxide (6%, 4%, 2%, 1%, 0.5%, 0.0%), then the amount of foam formed with highest volume will be in the cylinder with the most concentrated solution. The measuring cylinder with 6% hydrogen peroxide will produce the most foam as there is a lot more substrate for the enzyme to catalyze when it reacts with the fresh liver for 30 seconds. The enzyme catalase in fresh liver can break down the substrate, hydrogen peroxide (H 2O2), a lot faster if the concentration of the solution is increased. This is because by using a given amount of an enzyme, the reaction will speed up if the substrate is more concentrated. So there is and will be a significant difference between the different hydrogen peroxide concentration solutions. As the concentration of the substrate increases, the rate of reaction increases because there are more substrates to bind to the active site of an enzyme. There is a point, where all the active sites of an enzyme is occupied which means that no matter what the concentration is, the rate of reaction would not increase anymore (Enzyme Substrate). Materials: Graduated cylinder (100 cm3 and 5 or 10 cm3) Stopwatch Pipette Forceps Knife Detergent Solution of hydrogen peroxide (6%, 4%, 2%, 1% and 0.5%) Fresh liver Distilled water (100 cm3) 1

Aayush Desai Method:

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1. Cut 6 cubes of liver each (approximately 1cm x 1cm x 1cm) using a knife and ruler; one cube per concentration cylinder 2. Use the small graduated cylinder and pipette to measure 4 cm3 of 6% hydrogen peroxide with two drops of detergent into a 100 cm3 graduated cylinder and swirl to mix it 3. Using the forceps, take one cube of liver and place it in the graduated cylinder. 4. After 30 seconds, record the volume the foam reaches in the cylinder 5. Repeat step 2-4 with the other 4 concentration solutions of hydrogen peroxide and distilled water mixture. Make sure you rinse the cylinders carefully between procedures 6. Repeat steps 2-5 two more times to get additional data Variables: Independent Variable: The different concentrations of hydrogen peroxide (6%, 4%, 2%, 1%, 0.5%, and 0.0%) Dependent Variable: The volume (in cm3 using the scale on the cylinder) of foam produced in the graduated cylinder due to the concentration of the solution (hydrogen peroxide)

Controls for Variables: Variable Time Method of Control Using a stopwatch to measure 30 seconds each time when the reaction occurs to know the limit that the foam reaches in the measuring cylinder. Using the one same fresh liver to cut out the 1 cm3 pieces Having the same person in the group doing the same job, whether it is pouring in the solutions or cutting the 1 cm3 pieces of liver, to help minimize the human error margin; so we keep the jobs the same throughout the experiment

Liver source and volume Job allocations

Hydrogen Peroxide source and volume Soap

Aayush Desai Data Collection & Presentation Qualitative Data: Before The liver was bright red in color The hydrogen peroxide solution has no reaction and no form of fizz During The color of liver started to fade away Once the liver is added to the solution, foam starts to form and rise up the cylinder

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After In the end you can see the liver as a pale red color The reaction with the formation of foam starts to slow down and the foam starts to settle

Quantitative Data: Table 1: Raw Class Data of Experiment Hydrogen Peroxide Solution (%) 6 4 2 1 0.5 0.0 (Distilled Water) 1 31 62 34 24 9 4 2 41 43 42 24 13 7 3 38 31 29 10 13 6 Volume of foam formed (cm3 1 cm3) Trial 4 5 6 7 22 30 34 32 40 48 32 61 22 30 29 42 12 22 19 14 10 11 12 9 6 4 7 7

8 31 64 38 20 8 6

9 47 49 46 15 11 6

Table 2: My groups data of the Experiment Hydrogen Peroxide Solution (%) 6 4 2 1 0.5 0.0 (Distilled Water) Volume of foam formed (cm3 1 cm3) Trial 1 2 31 32 62 61 34 42 24 14 9 9 4 7

Aayush Desai Data Processing Table 3: Average of the class data Hydrogen Peroxide Solution (%) Average (cm3 1.0) 6 34 4 48 2 35 1 18 0.5 11 0.0 (Distilled water) 6 To calculate the average: plus all the data points / number of sample sizes For example: (64+43+31+40+48+32+61+64+49) / 9 = 48 cm3 Uncertainty (64+43+31+40+48+32+61+64+49) = 432 9 cm3 432 9 cm3 / 9 % uncertainty = 9/432 x 100 = 2.083333333 = 2.08% 432 2.08% / 9 = 48 2.08% / 100 x 48 = 0.9984 = 1.0 So, 48 1.0 cm3

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Standard Deviation Using Excel, standard deviation is calculated by: SD = STDEV (range) For example SD = STDEV (64, 43, 31, 40, 48, 32, 61, 64, 49) SD = 12.52775 2 sig fig: 12.5

Table 4: Standard Deviation of my groups data Hydrogen Peroxide Solution (%) 6 4 2 1 0.5 0.0 (Distilled water) Volume of foam formed (cm3 1 cm3) Trial 1 2 31 32 62 61 34 42 24 14 9 9 4 7 Standard Deviation 0.7 0.7 5.7 7.1 0.0 2.1

Aayush Desai Table 5: Standard Deviation of class data Hydrogen Peroxide Solution (%) 6 4 2 1 0.5 0.0 (Distilled Water) Data Presentation Figure 1: Class Average Graph 1 31 62 34 24 9 4 Volume of foam formed (cm3 1 cm3) Trial 2 3 4 5 6 7 8 41 38 22 30 34 32 31 43 31 40 48 32 61 64 42 29 22 30 29 42 38 24 10 12 22 19 14 20 13 13 10 11 12 9 8 7 6 6 4 7 7 6

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Standard Deviation 9 47 49 46 15 11 6 7.2 12.5 7.9 5.2 1.8 1.2

Class Average
60 50 40 30 20 10 0

Average (cm3 0.1)

Figure 2: Graph of Enzyme-substrate activity (Introduction to Enzyme Kinetics)

Aayush Desai Conclusion & Evaluation Conclusion:

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The hypothesis states that foam will form the most at the greatest volume (cm3) in the cylinder with the most concentrated hydrogen peroxide solution; this is moderately supported by figure 1. From the results in the data tables and figure 1, we can see that the cylinder with the 6% concentration of hydrogen peroxide didnt have the most amount of foam formed after 30 seconds. Looking at table 3, we can see that the averages clearly show that the cylinder with 4% hydrogen peroxide had the greatest amount of foam formed. The relationship we can see is that the foam increases according to the increased amount of percentage of the concentration. Table 3 does show that the volume increases (6cm3, 11cm3, 18cm3, 35cm3 and 48cm3) and the 6% concentration has an average volume of 34cm3. Figure 1 also supports this since we can see the blue line rises up, and then drops at 6%. So this result could in a way support the null hypothesis but the rest of the data and the graph prove that there are differences in the final outcome of the amount of foam produced. The results from the graph show that there is a positive difference between the concentration and amount of foam formed; from this we could reason that the results are directly proportional which means that the affect of one factor affects the affect of another. As we do not have enough trials or data we arent able to support the hypothesis thoroughly and to make a conclusion. Enzymes catalyze chemical reactions involving substrates. As the concentration of the substrate increases, the rate of reaction increases because there are more substrates to bind to the active site of an enzyme. As said earlier in the hypothesis, there is a point, where all the active sites of an enzyme is occupied which means that no matter what the concentration is, the rate of reaction would not increase anymore (Enzyme Substrate). If we look at figure 2, we can see the enzymesubstrate graph where the line reaches the point where the rate of reaction is stable and that is when all enzymes are occupied.

Aayush Desai Evaluation: Error Significance

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Size of Liver

High

Conditions of Liver

High

Stopwatch

Low

Different amounts of soap

Low

Lack of data

High

Improvement Instead of measuring the height, length and width to approximately 1cm3, by using the digital balance we can reduce the uncertainty since we can now measure the weight. From this we can make sure that all samples have the same weight rather than same approximate size. The experiment does take time so the liver cubes that were frozen when we started, started to melt as we went through the experiment. The melting of these liver cubes will affect the surface area which leads to different number of enzymes. We should find a way to keep the temperature or condition of the livers the same so maybe work in a freezer room, or wait for all liver cubes to melt and be at the same room temperature. There are different stopwatches and we are prone to human error when using them. This is because when we press stop for the different trials, there will be a small margin of milliseconds that are different. Soap tends to create random amounts of air bubbles and these bubbles can change the overall amount when mixing with concentration solutions. One reason could be that we should dilute the soap so that there is less uncertainty overall. This is very important because having more data can prove our hypothesis correct. In this case we needed to the experiment a few more times to find the actual outlier or the actual point where it shows the stability of the enzyme-substrate graph.

Bibliography: "Enzyme Substrate." WKU Biology Department. Web. 07 Nov. 2010. <http://bioweb.wku.edu/courses/biol115/Wyatt/Biochem/Protein/Active site.htm>. "Introduction to Enzyme Kinetics." D. W. Brooks Site. Web. 07 Nov. 2010. <http://dwb4.unl.edu/Chem/CHEM869K/CHEM869KLinks/www.curvefit.com/introduction63.h tm>. 7