Biochemistry Lec. of Biochemistry Dr. Mahmoud H.

Hadwan

Q1/ Calculate Km and Vmax from the following data:

[S] (M) 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 M) , the following initial velocities have

been reported

depending on the substrate concentration:

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Biochemistry Lec. of Biochemistry Dr. Mahmoud H. Hadwan

(a) Draw a Michaelis-Menten plot for this enzyme.

(b) Draw a Lineweaver-Burke plot for this enzyme.

(c) Determine KM and Vmax for this enzyme

(d) Indicate in both graphs (a & b) where Vma and KM can be

recognized.

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 0.1 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?

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Biochemistry Lec. of Biochemistry Dr. Mahmoud H. Hadwan

Q5/ An experiment was carried out using a food enzyme and gave the following
results.

1. plot Lineweaver-Burke for the data is reproduced below.

2. Comment on the resulting graphs

3. Estimate values of Vmax and Km for this data.

Q6/ The following data were obtained from an enzyme kinetics experiment.
Graph the data using a Lineweaver-Burk plot and determine, by inspection of the
graph, the values for Km and Vmax.
[S] (M) V (nmol/min)

_______ ___________

1.20 1.43
1.26 1.67
1.33 2.08
1.1 3.33

Q7/ Use the Michaelis-Menton Equation to calculate the missing values of [S]
given below if Vmax = 5 mmol/min. Plot [S] versus V (NOT the reciprocals!). Draw
line parallel to the x-axis at Vmax and extend your plotted line to show its
approach to Vmax.

[S] (mM) V (mmol/min)

_______ ___________

10 1.2
? 1.7

? 2.1

? 2.2

? 2.5

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Biochemistry Lec. of Biochemistry Dr. Mahmoud H. Hadwan

Q8/ The effect of an inhibitor on an enzyme was tested and the experiment gave
the results below. Plot the data and determine, by inspection of the graph, what
type of inhibition is involved.

[S] M V (mol/min)V (mol/min)V (mol/min)

with 0.0 nM with 25 nM with 50 nM

Inhibitor Inhibitor Inhibitor

______ ___________ ___________ ___________

0.4 0.22 0.21 0.20

0.67 0.29 0.26 0.24

1.00 0.32 0.30 0.28

2.0 0.40 0.36 0.32

Q9/ The following data were obtained from an enzyme kinetics experiment.
Graph the data using a Lineweaver-Burk plot and determine, by inspection of the
graph, the values for Km and Vmax.

[S] (M) V (nmol/min)

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Biochemistry Lec. of Biochemistry Dr. Mahmoud H. Hadwan

_______ ___________

1.21 1.43
1.27 1.67
1.34 2.08
2.1 3.33

Q10/ Use the Michaelis-Menton Equation to calculate the missing values

of [S] given below if Vmax = 5 mmol/min. Plot [S] versus V (NOT the
reciprocals!). Draw line parallel to the x-axis at V max and extend your
plotted line to show its approach to V max.

[S] (mM) V (mmol/min)

_______ ___________

11 1.2
[S]1 1.7

[S]2 2.1

[S]3 2.2

[S]4 2.5

Q11/ The effect of an inhibitor on an enzyme was tested and the

experiment gave the results below. Plot the data and determine, by
inspection of the graph, what type of inhibition is involved.

[S] M V (mol/min)V (mol/min)V (mol/min)

with 0.0 nM with 25 nM with 50 nM

Inhibitor Inhibitor Inhibitor

______ ___________ ___________ ___________

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Biochemistry Lec. of Biochemistry Dr. Mahmoud H. Hadwan

0.4 0.22 0.21 0.20

0.67 0.29 0.26 0.24

1.00 0.32 0.30 0.28

2.00 0.40 0.36 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