Caitlin Jade Esparza Lab # 5 9/28/11

Lab #5: Cellular Respiration Introduction: We need fuel for our body to function; we get this fuel from the food we eat in the form of proteins, carbohydrates, and fats. The carbohydrate glucose is the preferred energy source in our bodies. In order for this energy to be used, it must be broken down and transformed into ATP. Cells split glucose molecules to transfer the energy to ATP through a process called cellular respiration. In the first stage, a small amount of ATP is produced when glucose is broken down to pyruvate during glycolysis. In the second stage, the pyruvate either passes through aerobic respiration (large amount of ATP) or anaerobic fermentation (small amount of ATP). In the absence of oxygen, pyruvate is converted to either lactate or ethanol and carbon dioxide. Hypothesis: The germinating peas will have a higher rate of respiration than the beads and non-germinating peas. Materials: This experiment called for 20 germinating peas, 20 dry peas, 25 glass beads, 6 mL of 15% potassium hydroxide, 2 waterbaths, 6 respirometers (glass vials, graduated pipets, one-hole stoppers, washers), 6 absorbent cotton balls, 6 nonabsorbant cotton balls, 1 graduated cylinder (100 mL), 1 thermometer, 1 pipet, 1 glass marking pen, 1 stopwatch, ice, masking tape, petroleum jelly, food coloring, and paper towels. Methods: First, set up an ice waterbath, keeping the temperature at or right below 10 degrees Celsius. Obtain 6 vials with steel washers on the bottoms and number them 1-6. Fill a 100 mL graduated cylinder with 50 mL of water, then add 10 germinating peas and record the water displacement as the volume. Empty the water, pat the peas dry and set aside. Refill the graduated cylinder with 50 mL of water add 10 DRY NON germinating peas, then add glass peas until the water level is the same as that of the water with germinating peas. Again, remove the peas and beads and pat dry. Repeat the process with more germinating peas, non-germinating peas, and beads for vials 4-6. Place an absorbent cotton ball, not the non-absorbent rayon, in each of the six vials and push to bottom with a pipet or pencil. Add 1 mL of the KOH to the cotton and then place a slightly smaller piece of the non-absorbent rayon on top of the KOH soaked cotton in each vial. Using the first sets of germinating peas, non-germinating peas and glass beads, and glass beads, place them in vials 1-3, respectively. Do the same for vials 4-6. Insert the nontapered into the wide end of a stopper so that it extends just beyond the bottom of the stopper, do this for all 6 vials. Insert the stopper into the vial; this should be tight enough to prevent the pipet form moving around. Place a washer over the pipet tip and guide it down onto the top of the stopper,, again do this for all 6 vials. Now, place vials 1-3 in the room-temperature waterbath with the pipet edges resting on the edge of the tray. Place vials 4-6 in the chilled waterbath in the same manner. Let the vials sit (to equilibrate) for ten minutes. Now place a drop of food coloring on the exposed tip of each respirometer and wait a minute, then turn them so that the graduation marks on the pipets are

Caitlin Jade Esparza Lab # 5 9/28/11

facing up. Now immerse all respirometers in their respective waterbaths, do not touch the respirometers once the experiment has started. Let the respirometers equilibrate for another 5 minutes. Read all the respirometers to the nearest .01 mL and pay attention to the temperature of the bath. Take additional readings every 5 minutes for 30 minutes. When all the readings have been taken, calculate the difference and corrected difference for each result. Record all data on Table 1. Results: The results are shown on the Table 1. For vials 1-3, those in the room temperature bath, the readings went, from 0-30 min on 5 minute intervals, as .88, .83, .78, .76, .71, .68, and .65, respectively. The differences, the amount of water decreased from the original measurement, started at .05 at 5 minutes and ended with .23 at 30 minutes. These numbers are corrected for environmental factors and end up actually beginning at .01 at 5 minutes, and ending with .17 mL extra water after 30 minutes. This decreasing pattern is shown through all the vials at all temperatures. However, in the dry peas and beads vials at room temperature, water stopped decreasing after 15 minutes. The beads at room temperature didnt see and decrease in water after just 10 minutes. In the ice baths, oxygen kept being consumed. The reason that the ice baths continued to see a change, is because at the lower temperature, it took longer for cellular respiration to occur, so after 30 minutes, they were still working at cellular respiration, and therefore consuming oxygen, whereas at room temperature, the process was over. Also, it should be noticed that the beads are relatively consistent when compared to the vials with peas, the same ice-room temperature effect is present however. 1) According to your graph, what happens to the rate of oxygen consumed by germinating peas over time? What does this indicate to you? The rate of oxygen consumption is relatively constant, increasing constantly that is, as the slopes of the lines dont seem to vary much. 2) List at least three controls in this experiment. the temperatures of the ice baths, the amount of KOH and cotton balls, and the time are controls in this experiment. 3) Explain why the water initially moved into the respirometer. It is normal for a small amount of water and food coloring(although we did not use food coloring) to enter the pipet when first immersed because they are not tightly enough sealed to keep it out. 4) What is the role of the KOH in this experiment? KOH provides the oxygen and hydrogen needed for the oxidation by removal of hydrogen atoms and eventual combination with oxygen and hydrogen to form water in the electron transport system. KOH essentially takes the place of glucose. 5) How did the KOH affect the water movement in the respirometer? It allowed more water into the pipet.

Caitlin Jade Esparza Lab # 5 9/28/11

6) Which of the two pea types, germinating or non-germinating, consumes the most oxygen? Why? The germinating peas consume more oxygen because they are growing and need to consume more. 7) What was the effect of temperature on pea respiration? An increased temperature increased the rate of cellular respiration. 8) During aerobic respiration, glucose is broken down to form several end products. Which end products contain the carbon atoms from glucose? The hydrogen atoms from glucose? The oxygen atoms from glucose? The energy stored in the glucose molecules? Carbon dioxide contains the carbon, water contains the hydrogen, both carbon dioxide and water contain the oxygen, and ATP contains the energy. 9) What is fermentation? What are the two types of fermentation? What organisms use fermentation? Fermentation is a catabolic process that makes a limited amount of ATP from glucose without an electron transport chain and that produces an end-product such as ethyl alcohol or lactic acid. The two types of fermentation are alcoholic and lactic acid fermentation. Plants use alcoholic while animals use lactic acid. 10)

11) What are the three pathways involved in the complete breakdown of glucose to carbon dioxide and water? What reaction is needed to join two of these pathways? What are the substrates and products of this reaction and where does it take place? The three pathways are glycolysis, the electron transport chain, and the Krebs Cycle. The reaction of the pyruvate joining with CoA enzyme and NAD to produce acetyl CoA, NADH, and carbon dioxide. The acetyl CoA goes to the Krebs Cycle and NADH to the electron transport chain in the mitochondria

Caitlin Jade Esparza Lab # 5 9/28/11

12) Write the letter of the pathway that best fits each of the following processes. a. glycolysis b. Krebs Cycle c. Electron transport System 2) Water is formed C 1) Carbon Dioxide is given off B 3) PGAL A 4) NADH becomes NAD+ C 5) Oxidative Phosphorylation C 6) Cytochrome carriers C 8) FAD becomes FADH2 B 7) Pyruvate A 13) Calculate the energy yield of glycolysis and cellular respiration per glucose molecule. Distinguish between substrate-level phosphorylation and oxidative phosphorylation. Where does the energy for oxidative phosphorylation come from? 36 ATPs are formed per glucose moelcule. Four of the ATPs are formed from substrate level and 32 from oxidative. Glycolysis substrate(2) + oxidative(4)= 6 Transition substrate(0) + oxidative(6)= 6 Krebs substrate(2) + oxidative(22)= 24 Total substrate(4) + oxidative(32)= 36 14)

15) Dear Substitute, The body needs energy to function, the majority of this energy is held in the form of ATP (adenine triphosphate), which is largely formed through a process called cellular respiration. This is the process during which body cells split glucose molecules to transfer the energy to ATP. In the first stage, a small amount of ATP is produced as glucose is broken down to pyruvate through glycolysis. In the second stage, the pyruvate either passes through aerobic respiration (produces lg. amt. of ATP) or anaerobic fermentation (small amt. ATP). In eukaryotic cells, aerobic respiration occurs in the mitochondria, in prokaryotic cells, this occurs in the cell membrane. The overall equation is: Glucose + Oxygen Carbon Dioxide + Water + Energy (ATP). Sincerely, A Concerned Student 16) Name some other biological processes that are affected by temperature. Gas exchange, perspiration, heart rate, metabolism.

Caitlin Jade Esparza Lab # 5 9/28/11

Error Analysis: In the experiment, the original data for the ice bath vials was not accurate, saying that oxygen was consumed and then produced as the water level dropped and then increased. the reason for this error is most likely due to leaking of water into the vials at the colder temperature, which causes materials to shrink. In addition to this there were probably pressure changes with the water leakage. Discussion and Conclusion: This lab showed that germinating peas consumed more oxygen at a faster rate than the nongerminating peas and the beads did. The non-germinating peas and the beads didnt consume hardly any oxygen at all. It also showed that the respiration rate of germinating peas was faster than the respiration rate of non-germinating peas. Finally, this experiment showed that respiration rates increase as the temperature increases. This shows that temperature and respiration rates are directly proportional to each other.