Enzyme Kinetics and Mitochondrial Activity
Added on 2020-05-28
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1BIOSCIENCEBy NameCourseInstructorInstitutionLocationDate
2Electron transport and enzyme kinetics: succinate dehydrogenase activity in cauliflowerIntroduction.Studies have shown that in the eukaryotic organisms, there is cellular respiration process,which takes place in the mitochondria. During that process, amino acids, carbohydrates andfatty acids are all broken down to produce water and carbon dioxide. The energy releasedduring this reaction is either released as heat, transferred to molecules like the adenosinetriphosphate (ATP) or is used in the reduction of molecules such nicotinamide dinucleotide(NAD+) and the molecule Flavin adenine dinucleotide ( FAD)[CITATION Joh16 \p 345 \l 2057 ].Some of the mentioned reactions occur in the Krebs cycle, because of electronic transportchain and the rest occurs in the pathway of biochemical, which feeds intermediates into theKrebs cycle. The succinate dehydrogenase (SDH) is usually located in the innermost of themitochondrial membrane and most cases; it is the enzyme involved in this reaction ofcatalysing redox reactions, which passes two electrons from the succinate to the Flavinadenine dinucleotide and the FADH at the same time forming fumarate fromsuccinate[CITATION Nel12 \p 67 \l 2057 ]. Both the NADH and FADH have the capability of feeding electrons into many redoxreactions in the innermost of the mitochondrial membrane, mediated by the electron transportchain. At the finishing point of the molecular transport chain[CITATION Ath12 \p 345 \l 2057 ].The electrons in most cases are feed into oxygen molecules, which are combined and reducedwith two protons to produce water. In the case where the typical path of the electron transportis blocked by uncouples elements such as the potassium cyanide or Azide, or if oxygenmolecules are removed[CITATION Pat13 \p 432 \l 2057 ]. Artificial electron acceptors such as dichlopenolindophenol or the methylene blue applied toindicate the activities of the electron transport. That can be attributed to their oxidised state
3they are blue, but in the reduced state, they are colourless. This study uses this impact todetermine SDH activity in the cauliflower curds mitochondrial[CITATION Col13 \p 345 \l 2057 ].Aim/Objectives of the experimentThe objectives of this experiment to be carried out was to; Carry out mitochondrial isolation,measure succinate dehydrogenase activity, examine the viability of the electron transportsystem and finding out the effects which the competitive inhibitors have on the rate ofreaction. Experiment sectionMaterials and equipment.The materials used, equipment and chemicals for the practical included;Pestle and mortar, sharp sand, spatula, scalpel or razor blade, fresh cauliflower, ignitiontubes, spectrophotometer, cuvettes, cooled ultracentrifuge, pipettes, groves, pasture pipettes,paraffin, ice, scissors, cuvette stands and ignition tube racks.For the reagents, the following were used; Grinding buffer (0.3 M mannitol, 0.006 MKP2PO4, 0.014 MKHPO4 –PH 7.2) assay buffer, 0.04 M Azide, dichlorophenolindophenoland different concentrations of sodium succinate (0.2, 0.02, 0.002 and 0.0002)Procedure/methodThe groups divided themselves into two to carry out the following procedures.Isolation of the mitochondriaA cauliflower was taken and 25 grams of the outer surface removed.
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