Topic
Determining the Properties of an Enzyme
Instructions
Write a Biological report paper on Determining the Properties of an Enzyme
Answer Preview
Constant temperature water baths, spectrophotometers, computers with jmol program and peroxidase.pdb files, Mortar and pestle, Horseradish root, spectrophotometer cuvettes, 10ml test tubes and rack, 50ml beakers, 5ml pipettes graduated in 0.1 ml units with suction devices, thermometer, 10 mM H202, 25 mM guaiacol, 2% Hydroxylamine, citrate-phosphate buffers at PHs 3, 5, 7, and 9, peroxidase (horseradish), 6.4 mg/L in PH5 buffer if using sigma chemical source.
In order to determine whether the amount of enzyme influences the rate of reaction, test tubes were numbered 1-7. Test tube 1 contained all reactants but no enzymes to be used as a blank in calibrating the spectrophotometer. Test tube 2 contained a dilute concentration of peroxide while test 3 contained substrate and dye. Test tube 4 contained substrate and dye (same as test tube 2) while test tube 5 contained a medium concentration of peroxidase. Test tube 6 contained substrate and dye (same as test tube 2) while test tube 7 contained concentrated peroxidase. The content of test tube 2 and test tube 3 was mixed by pouring them back and forth twice for 10 seconds. The reaction mixture was quickly added to a cuvette by using an eyedropper.
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