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Botany Lab Report On Cellular Respiration.

2044 words - 8 pages

IntroductionAerobic respiration is the process by which mitochondria consume oxygen and organic fuel to produce ATP for the organism to use. Fermentation is another source of ATP for cells, but this method is by far, less effective in its yield of ATP. Glucose is the main source of organic fuel for cellular respiration, which releases as many as 38ATP molecules per glucose molecule in aerobic respiration. Unlike fermentation, which only releases 2 ATP molecules per glucose molecule.The first step in the process of aerobic respiration is glycolysis, which produces two molecules of pyruvate per cycle using one glucose molecule. Pyruvate is then used in the Krebs cycle to release electrons in certain steps. These electrons then enter the electron transport chain where a series of steps produce the majority of the ATP formed during aerobic respiration.Glycolysis takes place in the cytoplasm, while the other two steps occur within the mitochondrion organelle. In order to study the reaction of aerobic respiration, a source of mitochondria is needed. Using a microcentrifuge, the mitochondria are removed from a solution of ground up lima beans. The mitochondria still carry out aerobic respiration even when they are removed from the rest of their cell contents. The source of glucose for this suspension is sucrose. Sucrose is processed readily by the mitochondria as it is a rich source of the sugar glucose.To measure the rate of cellular respiration, you need to monitor the activity of enzymes in the system that are involved with the Krebs cycle. In one step, succinate is converted to fumarate. During this step, electrons are released from the reaction, which travel to the electron transport chain where they are used to make ATP.In this experiment, DPIP is a blue dye used to indicate the presence and level of electrons in the solution of mitochondria before the electrons reach the electron transport chain. When a DPIP molecule grabs an electron, its color changes from blue to colorless. The degree of color change indicates the level of enzymatic activity occurring and using a spectrophotometer, the color change can be monitored and recorded. The transmittance level of the sample will indicate how much enzymatic activity is occurring.Electrons are released gradually down the electron transport chain. This is the stage where ADP is converted to ATP. DPIP acts as a substitute for ubiquinone, the only lipid in the transport chain. Since DPIP has a greater affinity for electrons than ubiquinone, it accepts the electrons and becomes reduced and colorless before the electrons reach the bottom of the chain.Using these methods, we are able to observe the level of aerobic respiration occurring within the mitochondria of living cells.MethodsA. Mitochondria Isolation.1.Use a scalpel to cut soaked lima beans into pieces. (The lima beans must be soaked for about 12 hours in water before cutting them up.) Grind the cut seeds with 25 mL of assay buffer for two minutes in...

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