1. Lab Session 8 IUG, Spring 2015 TMZ IUG, Spring 2015 TMZ 1

2. Energy Metabolism 2

3. Cellular Respiration Aerobic 1.Krebs cycle 2.Electron Transport Chain Anaerobic 1.Alcoholic Fermentation 2.Lactate Fermentation Aerobic 1.Krebs cycle 2.Electron Transport Chain Anaerobic 1.Alcoholic Fermentation 2.Lactate Fermentation 3

4. Cellular Respiration Stages of Cellular Respiration and Fermentation 4

5. The Route of Glucose Metabolism 5

7. Experiment Alcoholic Fermentation in Yeast Reagents & Instruments 5 respirometers consisting of: pipette pump test tube graduated pipettes 1-ml graduated pipette yeast solution aquarium tubing glucose solution 250 ml flask distilled water binder clips sharpie Alcoholic Fermentation in Yeast Reagents & Instruments 5 respirometers consisting of: pipette pump test tube graduated pipettes 1-ml graduated pipette yeast solution aquarium tubing glucose solution 250 ml flask distilled water binder clips sharpie Respirometer used for yeast fermentation 7

8. Procedure 1. Obtain 5 flasks and fill with approximately 200 ml of tap water. Label the flasks 1, 2, 3, 4, and 5. Place the water filled flasks into separate beakers (this should stabilize the respirometers). 2.Obtain 5 test tubes (fermentation tubes) and label them 1, 2, 3, 4, and 5. Add solutions as follows to the appropriate tubes. Procedure 1. Obtain 5 flasks and fill with approximately 200 ml of tap water. Label the flasks 1, 2, 3, 4, and 5. Place the water filled flasks into separate beakers (this should stabilize the respirometers). 2.Obtain 5 test tubes (fermentation tubes) and label them 1, 2, 3, 4, and 5. Add solutions as follows to the appropriate tubes. Tube Water (ml) Yeast Suspension (ml) Glucose Solution (ml) 1 4 0 3 2 6 1 0 3 3 1 3 4 1 3 3 5 2 2 3 8

9. 3.Attach a piece of aquarium tubing to the end of each 1-ml graduated pipette. Then place the pipette with attached tubing into each test tube containing fermentation solutions. 4.Attach the pipette pump to the free end of the tubing on the first pipette. Use the pipette pump to draw the fermentation solution up into the pipette. Fill it past the calibrated portion of the tube, but do not draw the solution into the tubing. 5. Fold the tubing over and clamp it shut with the binder clip so the solution does not run out. Open the clip slightly, and allow the solution to drain down to the 0-ml calibration line (or slightly below). Quickly do the same for the other four pipettes. *Note: if you can not get your solution to the 0-ml mark, then just do your best, mark the starting point with a sharpie and note your initial reading in Table 2. 3.Attach a piece of aquarium tubing to the end of each 1-ml graduated pipette. Then place the pipette with attached tubing into each test tube containing fermentation solutions. 4.Attach the pipette pump to the free end of the tubing on the first pipette. Use the pipette pump to draw the fermentation solution up into the pipette. Fill it past the calibrated portion of the tube, but do not draw the solution into the tubing. 5. Fold the tubing over and clamp it shut with the binder clip so the solution does not run out. Open the clip slightly, and allow the solution to drain down to the 0-ml calibration line (or slightly below). Quickly do the same for the other four pipettes. *Note: if you can not get your solution to the 0-ml mark, then just do your best, mark the starting point with a sharpie and note your initial reading in Table 2. 9

10. 6.Record your initial readings for each pipette. This will be the initial time (I). 7.2 minutes after the initial readings for each pipette, record the actual readings (A) in ml for each pipette in the “Actual (A)” column. 8.Subtract I from A to determine the total amount of CO 2 evolved (A-I). Record this value in the “C0 2 Evolved (A-I)” column. From now on, you will subtract the initial reading from each actual reading to determine the total amount of CO 2, evolved. 9.Continue taking readings every 2 minutes for each of the solutions for 20 minutes. Remember, take the actual reading from the pipette and subtract the initial reading to get the total amount of CO 2 evolved in each test tube. 6.Record your initial readings for each pipette. This will be the initial time (I). 7.2 minutes after the initial readings for each pipette, record the actual readings (A) in ml for each pipette in the “Actual (A)” column. 8.Subtract I from A to determine the total amount of CO 2 evolved (A-I). Record this value in the “C0 2 Evolved (A-I)” column. From now on, you will subtract the initial reading from each actual reading to determine the total amount of CO 2, evolved. 9.Continue taking readings every 2 minutes for each of the solutions for 20 minutes. Remember, take the actual reading from the pipette and subtract the initial reading to get the total amount of CO 2 evolved in each test tube. 10

11. Tube 1Tube 2 Tube 3 Tube 4 Tube 5 CO 2 TimeActualEvolvedActualEvolvedActualEvolvedActualEvolvedActualEvolved (min)(A)(A-I)(A)(A-I)(A)(A-I)(A)(A-I)(A)(A-I) Initial Reading (I) 2 4 6 8 10 12 14 16 18 20