Calculate Km and Vmax from the following data.docx

Biochemistry Lec. of Biochemistry Dr. Mahmoud H.Hadwan Q1/ Calculate Km and Vmax from the following data: [S] (µM) V0 0.1 0.34 0.2 0.53 0.4 0.74 0.8 0.91 1.6 1.04 Q2/ Determine the type of inhibition of an enzymatic reaction form the follwoing data collected in the presecnce and absence of the inhibitor [S] [V0] V0 with I present 1 1.3 0.8 2 2.0 1.2 4 2.8 1.7 8 3.6 2.2 12 4.0 2.4 Q3/ For an enzyme (5 μM) , the following initial velocities have been reported depending on the substrate concentration: 1 1 mM? b) What is the initial reaction velocity at a substrate concentration of 30 mM? c) What is the initial reaction velocity at a substrate concentration of 1000 mM? 2 .Biochemistry Lec. of Biochemistry Dr. (b) Draw a Lineweaver-Burke plot for this enzyme. Q4/ Given the reaction of an enzyme that follows Michaelis-Menten kinetics: k1 kp E + S  ES  E + P k-1 If Km = 30 mM and Vmax = 60 uM min-1 a) What is the initial reaction velocity at a substrate concentration of 0. Mahmoud H. (c) Determine KM and Vmax for this enzyme (d) Indicate in both graphs (a & b) where Vma and KM can be recognized. Hadwan (a) Draw a Michaelis-Menten plot for this enzyme. 1 ? 2.5 3 .Biochemistry Lec. Draw line parallel to the x-axis at Vmax and extend your plotted line to show its approach to Vmax.7 ? 2. Comment on the resulting graphs 3. the values for Km and Vmax. [S] (mM) V (mmol/min) _______ ___________ 10 1.43 1. [S] (µM) V (nmol/min) _______ ___________ 1.2 ? 2.33 2. of Biochemistry Dr.2 ? 1. 1.26 1. plot Lineweaver-Burke for the data is reproduced below.08 1.20 1. Graph the data using a Lineweaver-Burk plot and determine. Mahmoud H. Plot [S] versus V (NOT the reciprocals!). 2.1 3. by inspection of the graph. Q6/ The following data were obtained from an enzyme kinetics experiment. Estimate values of Vmax and Km for this data.67 1. Hadwan Q5/ An experiment was carried out using a food enzyme and gave the following results.33 Q7/ Use the Michaelis-Menton Equation to calculate the missing values of [S] given below if Vmax = 5 mmol/min. Hadwan Q8/ The effect of an inhibitor on an enzyme was tested and the experiment gave the results below.32 0. Plot the data and determine.40 0. Graph the data using a Lineweaver-Burk plot and determine.67 0.22 0.0 nM with 25 nM with 50 nM Inhibitor Inhibitor Inhibitor ______ ___________ ___________ ___________ 0.20 0. the values for Km and Vmax. by inspection of the graph. of Biochemistry Dr.24 1.30 0.00 0.Biochemistry Lec.4 0. [S] µM V (µmol/min)V (µmol/min)V (µmol/min) with 0. by inspection of the graph. Mahmoud H.0 0.21 0.29 0. [S] (µM) V (nmol/min) 4 .28 2.36 0. what type of inhibition is involved.32 Q9/ The following data were obtained from an enzyme kinetics experiment.26 0. [S] µM V (µmol/min)V (µmol/min)V (µmol/min) with 0. what type of inhibition is involved. Mahmoud H.1 [S]3 2. by inspection of the graph. of Biochemistry Dr.5 Q11/ The effect of an inhibitor on an enzyme was tested and the experiment gave the results below.67 1.1 3.27 1.33 Q10/ Use the Michaelis-Menton Equation to calculate the missing values of [S] given below if Vmax = 5 mmol/min.2 [S]4 2. [S] (mM) V (mmol/min) _______ ___________ 11 1.21 1.Biochemistry Lec. Plot [S] versus V (NOT the reciprocals!).08 2. Hadwan _______ ___________ 1. Plot the data and determine.34 2.7 [S]2 2.0 nM with 25 nM with 50 nM Inhibitor Inhibitor Inhibitor ______ ___________ ___________ ___________ 5 .43 1. Draw line parallel to the x-axis at V max and extend your plotted line to show its approach to V max.2 [S]1 1. 30 0. of Biochemistry Dr.20 0.28 2.00 0.40 0.21 0. Mahmoud H.32 0. Hadwan 0.4 0.29 0.32 Additional Plot of Michaelis-Menton Kinetics Plot Lineweaver-Burke of COMPETITIVELY inhibited M-M reaction Plot Lineweaver-Burke of Uncompetitive Inhibition Plot Lineweaver-Burke with Mixed Inhibition 6 .00 0.26 0.24 1.67 0.22 0.36 0.Biochemistry Lec.