Study Of Krebs Cycle Using Mitochondria From Mung Bean Seedlings

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Experiment 4: Study of Krebs Cycle using Mitochondria from Mung Bean Seedlings Matthew Foley 100769849 THURS PM BIOL 2200 A04 Bench #2 Introduction In this experiment the functional mitochondria were isolated from etiolated mung bean hypocotyls through the processes of homogenization and centrifugation. The process of homogenization disrupts the cell tissue in order to release the contents of the cells into the homogenate. Once the organelles have been released into the homogenate they are isolated by centrifugation. This process is effective due to the fact that different cellular structures move through the homogenate differently, based mostly on size and density. Therefore, different centrifugal forces and times are used, specific to which organelle or cellular component you are trying to isolate. Mitochondria play a very important role in the cell since they generate most of the cell’s supply of ATP by the phosphorylation of ADP to ATP. This process involves a number of chemical reactions which together are referred to as cellular respiration. The first step is glycolysis which produces pyruvate and NADH from glucose. The pyruvate is then transferred to the mitochondria where they are converted into acetyl-coenzyme A. at this point the acetyl-coenzyme A enters the Krebs Cycle where it forms a series of intermediates. These intermediates then provide electrons in order to reduce the coenzymes NAD+ and FAD to NADH and FADH2 respectively. These electrons which are carried by NADH and FADH2 are then sent to the electron transport chain where they are eventually used to reduce O2 to H2O. The energy released from the electron transport chain is then used to phosphorylate ADP to ATP. The experiment performed follows the progression of the Krebs Cycle through the use of a dye called 2,6 dichlorophenolindophenol (DCPIP). DCPIP is a blue

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