Name: ___________ Rennin Enzyme Lab

Period: _____

Digestion is the breakdown of food into molecules useable by the body. Enzymes are chemicals that help digest food. Chymosin (rennin) is an enzyme that is found in the stomach of milk-producing animals. The enzyme leads to curd formation and aids in digestion. Procedures: Students will work in pairs. Each pair will test EITHER enzyme concentration, temperature OR pH. Part 1: Enzyme Concentration 1. Obtain six test tubes and label them 1 through 6. 2. Add 2 ml of whole milk to each tube. 3. Add the following amounts of enzyme to the test tubes using a calibrated plastic pipette. a. Tube 1 = 0 ml b. Tube 2 = 0.5 ml c. Tube 3 = 1 ml d. Tube 4 = 2 ml e. Tube 5 = 3 ml f. Tube 6 = 4 ml 4. Start timing as soon as the enzyme is added. 5. After adding the enzyme, put your thumb over the top of the test tube and mix. Do this approximately every 30 seconds. 6. Record the time it takes for the milk in each tube to begin to coagulate. Use the appropriate table provided. 7. Prepare a graph (X-axis amount of enzyme, Y-axis Minutes to coagulation). Part 2: Temperature 1. Obtain seven test tubes and label them 1 through 7. 2. Add 2 ml of whole milk to each tube 3. Add 1 ml of enzyme to each tube EXCEPT tube 1 4. After adding the enzyme, put your thumb over the top of the test tube and mix. Do this approximately every 30 seconds 5. Place the tubes at various temperatures as follows: a. Tubes 1 & 2 & 3 at room temperature b. Tubes 4 & 5 on ice c. Tubes 6 & 7 in a warm water bath. 6. Record the time for the milk in each tube to begin to coagulate. Use the appropriate table provided. 7. Prepare a graph (X axis = Temperature of milk, Y-axis = Minutes to coagulation).

Part 3: pH 1. Obtain eight test tubes and label them 1 through 8. 2. Add 2 ml of whole milk to each of eight test tubes. 3. Add 1 ml of each of the 7 vinegar solution to separate test tubes as follow: EX. Test tube 1 vinegar solution 1 pH=2.8 Test tube 2 vinegar solution 2 pH=3.0 Test tube 3 vinegar solution 3 pH=3.2 Test tube 4 - vinegar solution 4 pH=3.5 Test tube 5 - vinegar solution 5 pH=3.8 Test tube 6 - vinegar solution 6 pH=4.1 Test tube 7 - vinegar solution 7 pH=4.7 Test tube 8 distilled water pH=7.0 Test tube 9 distilled water pH=7.0 NO ENZYME Use ONLY the pipette that is in each solution. DO NOT mix the pipettes. 4. Add 1 ml of enzyme to test tubes 1-8. DO NOT ADD enzyme to test tube 9. 5. After adding the enzyme, put your thumb over the top of the test tube and mix. Do this approximately every 30 seconds 6. Record the time for the milk in each tube to begin to coagulate in the appropriate table provided. 7. Prepare a graph (X-axis = pH of milk, Y-axis = Minutes to coagulation) Clean Up: 1. Use a test tube brush to scrub out your test tubes. Rinse them well with water. Place them upside down in the test tube rack at your station. 2. Get data for the two sections your team did not do from other teams. 3. Graph the data on three separate graphs. Graph pH vs. time, enzyme concentration vs. time, and temperature vs. time.

Discussion questions 1. Describe the effect of Rennin on Milk.

2. Have you ever seen a baby spit up? Did it look anything like this? Why do you think it did?

3. Describe the effect of enzyme concentration on milk coagulation.

4. Describe the effect of temperature on milk coagulation.

5. Describe the effect of pH on milk coagulation.

6. What evidence do you have that the Rennin was responsible for the change in the milk? What do you call this element of the experiment?

7. Why would rennin be useful in making ice cream or cheese?

8. What is the difference between an enzyme and a catalyst?

9. Is rennin an enzyme or a catalyst? Why?

10) What other factors do you think could change the effect of the Rennin?

DATA TABLES Concentration Volume of Rennin (ml)

Tube 1 2 3 4 5 6

Coagulation Time (Minutes)

Tube 1 2 3 4 5 6 7

Temperature Temperature of Milk C

Coagulation Time (Minutes)

Tube 1 2 3 4 5 6 7 8

pH pH of Milk

Coagulation Time (Minutes)

GRAPHS

Name______________

Teacher Preparation Materials

Vinegar beakers roasters test tube racks rennet tablets Distilled Water plastic pipettes metal test tube racks test tubes test tube brushes pH probe aluminum foil test tube clamps labeling tape stop watches flasks Hot plates thermometers marker

Enzyme solution is made by mixing one rennet tablet with 50 ml of water. Need about 3 tablets per class of 26-30 students. Amount of shaking the test tube greatly influences rate of coagulation. If you are really going to use a class average, make sure method of shaking is established. Students work as a pair on only one of the tests, pH, concentration, or temperature. Students then get the data for the other two tests from other groups. I found it best to have students go into the lab and depending on where they were standing they were placed into their test groups.

Temperature Notes:
5 samples (one cold from the refrigerator, one room temperature, three warmed in water bath). Only warmed ones will coagulate within the class period.

Concentration Notes: pH Notes:

3 ml + will coagulate within the class period.

pH solutions of 2.8 and sometimes 3.0 coagulate milk without enzyme! Be sure to cover with aluminum foil to minimize evaporation. Also, it is important that the plastic pipettes are not transferred from one solution to another. The pH solution of 4.7 required only 0.1 ml of vinegar in 100 ml of water, so transferring even a small amount of vinegar can affect the pH greatly! Other substances that could be tried could be citric acid (ph ~2.4, baking soda, ammonia, etc.) pH solutions were prepared as follows: Volume of vinegar(ml) Volume of water(ml) pH (using probe) 0 100 7.0 0.1 100 4.7 0.25 100 4.1 1.0 100 3.8 3.0 100 3.5 8.0 100 3.2 30 100 3.0 100 0 2.8