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Laboratory Report
Name: Ye Mon Tun Title: Catalytic Activity of Enzymes
1. 2. 3. 4. 5. Introduction Materials and methods Results Discussion Conclusion

Introduction Def: Enzymes are proteins that catalyze chemical reactions. In enzymatic reactions, the molecules at the beginning of the process are called substrates, and the enzyme converts them into different molecules, called the products. Almost all processes in a biological cell need enzymes to occur at significant rates. Since enzymes are selective for their substrates and speed up only a few reactions from among many possibilities, the set of enzymes made in a cell determines which metabolic pathways occur in that cell. In enzymatic reactions, the molecules at the beginning of the process are called substrates, and the enzyme converts them into different molecules, called the products. Almost all processes in a biological cell need enzymes to occur at significant rates. Since enzymes are selective for their substrates and speed up only a few reactions from among many possibilities, the set of enzymes made in a cell determines which metabolic pathways occur in that cell.

Several factors that are especially important in influencing enzyme activity are: 1. Enzyme concentration 2. Substrate concentration 3. Salt concentration 4. PH 5. Temperature Materials and Methods Potato Extract 3% H2O2 solution 1% H2O2 solution pH3, pH5, pH7, pH9 and pH11 buffer solution Distilled water Filter paper disks Forceps 100ml beakers Measuring cylinder Stop watch Methods 1. The effect of enzyme concentration on enzyme activity a) Label the beakers: 20%, 40%, 60%, 80% and 100%. b) Make the appropriate dilutions with the potato cell extract as following table Test Enzyme Volume of Potato Volume of Water Concentration Cell Extract (ml) (ml) #1- Control 0% 0 40 #2 20% 8.0 32.0 #3 40% 16.0 24.0 #4 60% 24.0 16.0 #5 80% 32.0 8.0 #6 100% 40.0 0 c) Pour 30 ml of the 1% hydrogen peroxide into a clean beaker. d) With forceps, pick up and dunk a paper disk in the enzyme for 5 seconds and then drain off. e) Place the disk into the bottom of the baker which contains 1% hydrogen peroxide. f) Using Stopwatch, record the time that the disk reaches the top of the liquid in a flat orientation. g) Repeat the above (step d to f) three times, each time with clean disk.

2. The effect of substrate concentration on enzyme activity a) Label the clean beakers with 2%, 1.5%, 1%, 0.8%, 0.6%, 0.3% and 0%. b) Make the appropriate dilution with water and 3% hydrogen peroxide as below table. Test Substrate Volume of 3% Volume of water Concentration hydrogen (ml) peroxide (ml) #1 2% 20 10 #2 1.5% 15 15 #3 1% 10 20 #4 0.8% 8 22 #5 0.6% 6 24 #6 0.3% 3 27 #7- Control 0% 0 30 c) Obtain 60% catalase enzyme solution. d) With forceps, pick up a paper disk, dunk in the 60% enzyme solution for 5 seconds and then drain off. e) Next, place it onto the bottom of the beaker containing 2% substrate concentration. f) Using Stopwatch, record the time that the disk reaches the top of the liquid in a flat orientation. g) Repeat this procedure for all the other substrate concentrations. 3. The effect of pH on enzyme activity a) Label the beakers: pH3, pH5, pH7, pH9 and pH11. b) Dilute the photo cell extract containing catalase as follows. Test #1-pH3 #2-pH5 #3-pH7 #4-pH9 #5-pH11 Potato Cell Extract (ml) 5 5 5 5 5 Respective pH Buffer 5 5 5 5 5

c) Mix and allow sitting for 5 minutes. d) Pour 30 ml of the 1% hydrogen peroxide into the clean beaker. e) Pick up a paper disk and dunk in the diluted catalase for 5 seconds. f) With forceps, place it onto the bottom of the baker containing 1% hydrogen peroxide.

g) the h) Result 1. Test

Using Stopwatch, record the time that the disk reaches the top of liquid in a flat orientation. Repeat this procedure for all diluted catalase extract at different pH. Data Table for Effect of Enzyme Concentration Trial 1 (seconds) 100 38 28 28 30 Trial 2 (seconds) 101 39 37 24 25 Trial 3 (seconds) 92 47 31 23 31 Average (seconds) 97.667 41.333 32 25

#1-Control #2 #3 #4 #5 #6

Test #1 #2 #3 #4 #5 #6 #7-

2. Data Table for Effect of Substrate Trial 1 Trial 2 (seconds) (seconds) 22 31 38 30 43 47 53 47 63 65 136 138 Control -

Concentration Trial 3 Average (seconds) (seconds) 30 27.333 36 34.667 47 45.667 55 51.667 70 66 142 139 -

3. Data Table for Effect of pH Test Trial 1 Trial 2 (seconds) (seconds) #1-pH3 #2-pH5 56 65

Trial 3 (seconds) 70

Average (seconds) 63.667

#3-pH7 #4-pH9 #5-pH11

24 19 29

25 20 31

23 23 30

24 20.667 30

Discussion