Enzyme Kinetic Lab Report

Abstract The main goal of the enzyme kinetics experiment was to see how the phosphatase-catalyzed hydrolysis of p-nitrophenyl produced p-nitrophenol in the presence of phosphate and fluoride ion inhibitors of various concentrations. The calculated Km constant was found to be 0.22 for all reactions. The Vmax values for each inhibition ion were 0.00986 for the phosphate ion and 0.00436 for the fluoride ion. The inhibitor constant, Ki, was determined to be 0.0967 for the phosphate ion. The inhibitor constant for the fluoride ion was determined to be 0.137. Both ion inhibitors resembled competitive inhibition.
Introduction
Enzyme kinetics is a classification of chemically catalyzed reactions that take place in the presence of an enzyme.6 Enzyme kinetic reactions evaluate the reaction rate of a given reaction under varying biological conditions.6 Enzymatic reactions are used to evaluate the catalytic mechanism of an enzyme and determine how it controls its activity.6 Enzymes come in the form of protein molecules that have the ability to interact and change the composition of the enzyme's
Diluted five small test tubes for each of the 3 sets of phosphate and fluoride ion concentrations with solutions and volumes that were pre-determined before the initiation of the lab.1 Placed 50uL of the enzyme in each test tube.1 Ran each reaction for the duration of five minutes.1 Quenched each test tube with 250uL of 6M NaOH after five minutes.1 Measured the absorbance of each sample at 405nm and recorded all data.1 Sodium hydroxide is corrosive and an irritant.3 Sodium phosphate is toxic and is a skin and eye irritant.4 Sodium fluoride is corrosive and is a skin and eye irritant.2 Triton X-100 is corrosive and may cause eye and skin irritation.5 All chemicals should be used in the presence of a laboratory instructor and should handled only when wearing gloves and protective