Enzymes are catalysts. They make things work faster. Enzymes are the key to life. No enzymes, no life.

We destroy all the digestive enzymes in our food by cooking and baking, and thus the body has to draw the necessary enzymes to digest this dead food from the body organs which in turn become unbalanced. All processed foods are also void of enzymes. Research has shown that this might be a cause of a lot of degenerative diseases. Fortunately eating raw food and/or taking enzyme supplements can help to restore our health. Enzymes serve as the labor force to perform every single function required for our daily activities and are required to keep us alive. Most of these substances are synthesized from components of ingested food, water, and other nutritional supplements, or from breakdown products of tissues. The biochemical reactions to synthesize the basic elements required by living cells depend on a steady supply of energy from these sources.

MATERIALS Potato Fehlings solution Iodine solution Hydrogen peroxide (3%) Buffer Solution 10% NaOH 10% HCl Cheesecloth Shredder /Grater Thermometer Mortar and pestle Bunsen and burner Starch solution Saliva Stirring rod Test tube Ruler

I. 1. 2. 3.

4. 5.

Activity of amylase Collect about 10 drops of your own saliva in a big test tube Add 6 drops of starch solution to the prepared saliva in the test tube. Add 3 drops of iodine solution and stir. The solution should be light blue. Record the time of formation of the blue color. Let the solution stand for a few minutes and record the time when the blue color disappears. Transfer 5 drops of the solution from step 4 to a test tube and add 2 ml of Fehling's solution. Place in water bath for 5 minutes. observe

II. Activity of Catalyst a. Extraction of Catalyst Peel and shred the potato with the grater. Soak with 25ml of water, and filter through cheesecloth.

b. Effect of temperature 1. Place 2ml of filtrate into a test tube. 2. Add H202 up to 3cm mark in the test tube. Stir 3. Observe the formation of bubbles by the action of the potato enzyme on the hydrogen peroxide. 4. Measure the height of the foam after 1 minute. Measure from the top of the liquid to the top of the foam. 5. Measure the foam height every minute for a total of 5 minutes. 6. Construct a graph of foam height on the y-axis versus time on the xaxis 7. Repeat B step no.1 with the following test tubes. Place each tube at a constant temperature for several minutes. test tube no.2 ice bath (0 Celsius) test tube no.3 room temp bath (20-25 Celsius) test tube no.4 body temp bath (37-40 Celsius) test tube no.5high temp bath (70-80 Celsius) 8. Remove the tubes from the baths and add H2O2 up to 3 cm in the test tube. put the tubes back into the bath. 9. Compare the rates of gas production by measuring the foam height in each tube for 5 minutes, similar to step no.5 10. Construct a graph, similar to step no.1 for each test tube.

c. Effect of pH
1. 2. 3. 4. 5. 6.

Place potato pulp in three test tubes to a depth of about 2 cm. label the test tubes 1,2, and 3. To test tube no. 2, add 5 drops of 10 % NaOH To test tube no. 3, add 5 drops of 10 % HCl Determine and record the pH of the solutions in the three test tubes. Add H2O2 up to 3 cm mark of the test tubes. Measure the foam height in each tube after 1 minute.

a.

Activity of amylase
Time 3min 6min 9min Blue color formation negative negative negative Disappearance of blue color negative negative negative

12min

negative

negative

b. Effect of Temperature
Time Test interval tube no. 1 (height in cm) 1 min 2 min 3 min 4 min 5 min 15 cm 15 cm 16 cm 16 cm 17 cm Test tube no.2 at 0 Celsius 3 4.5 6 7 8 Test tube no. 3 at 2025 Celsius 2.5 2.7 3.5 3.7 4 Test tube no. 4 at 3740 Celsius 2.5 2.7 3.5 3.7 4 Test tube no. 5 at 7080 Celsius 2.7 3.5 4.3 4.5 4.8

c. Effect of pH
Test tubes
No. 1 No. 2 No. 3 7 14 2

pH

Height in cm (foam)
16 cm 25 cm 10 cm

1. 2. 3. 4. 5. 6. 7.

What are the uses of starch and iodine solutions in the determination of amylase activity? How would you explain the disappearance of color of iodine with starch after a few minutes? What enzyme is present in potato? What is the effect of temperature on the action of potato enzyme? What is the effect of temperature on the action of hydrogen peroxide? Compare and account for the result in graph formed Compare and account for the effect of pH on the activity of potato enzyme with hydrogen peroxide

4. What is the effect of temperature on the action of potato enzyme? 5. What is the effect of temperature on the action of hydrogen peroxide? 6. Compare and account for the result in graph formed 7. Compare and account for the effect of pH on the activity of potato enzyme with hydrogen peroxide

1.) To measure your amylase activity, you will monitor the disappearance of amylases substrate, starch. Starch reacts with iodine (which is yellow) to form a blue compound (Amax 620 nm). This reaction is the basis of a colorimetric assay for amylase activity. Broth culture supernatant (which contains the secreted amylase) will be incubated with starch. After the incubation period, a portion (an aliquot) of this mixture is combined with acidic iodine. The acid stops the enzymatic reaction and the iodine reacts with the starch to produce the blue color. Any starch that has not yet been hydrolyzed by the amylase will turn blue, with the intensity of the blue color being proportional to the amount of starch remaining.

2.) Theres actually no disappearance of iodine with starch because theres no reaction recorded. 3.) Catalase is the most well known enzyme common in potato, but there are others enzymes that can be found in potato such as Oxidase and peroxidase.

4.) The higher the temperature of water, potato and HO, the rate at which the Enzyme will work will be faster therefore producing more oxygen. The reaction will be the same without the catalase (potato). Therefore in both experiments the Enzyme will work more rapidly and produce more oxygen.

5.) It increased the height of the foam formed in each test tube 6.) hydrogen peroxide increased the rate of chemical reaction of the potato as an enzyme