Last edited: 9/12/2021

1. MICHAELIS MENTEN & LINEWEAVER-BURK PLOT

Biochemistry | Michaelis Menten & Lineweaver-Burk Plot Medical Editor: Uta Hüning

OUTLINE (i) Competitive inhibition

𝑘𝑘𝑚𝑚 increases
I) GENERAL
II) MICHAELIS MENTEN CURVE 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 stays
III) LINEWEAVER-BURK PLOT
IV) APPENDIX
V) REVIEW QUESTIONS

I) GENERAL
Michaelis Menten curve
o Rectangular hyperbola
o According to Michaelis Menten equation
Lineweaver-Burk plot
o Linear form
o Inverse function of Michaelis Menten equation
Figure 2: Competitive inhibition in Michaelis Menten curve
II) MICHAELIS MENTEN CURVE (ii) Non-competitive inhibition
(1) Curve 𝑘𝑘𝑚𝑚 stays
𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 decreases

Figure 1: Michaelis Menten saturation curve

X- axis:
o Substrate concentration [𝑆𝑆]
o Course: Figure 3: Non-competitive inhibition in Michaelis Menten curve
 Increases over time
(iii) Uncompetitive inhibition
Y-axis:
o Velocity 𝑉𝑉0 𝑘𝑘𝑚𝑚 decreases
o Course: 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 decreases
 Beginning: proportional to substrate concentration
 Later: constant
(2) What happens?
Enzyme reacts with substrate

Linear part:
o Substrate concentration increases → rate of reaction
increases proportionally

 Meaning: 𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟 𝑜𝑜𝑜𝑜 𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟 = 𝑘𝑘 ∗ [𝑆𝑆]𝟏𝟏 = 𝑘𝑘 ∗ [𝑆𝑆]

Plateau part:
o More substrate is added but velocity doesn’t increase
anymore Figure 4: Uncompetitive inhibition in Michaelis Menten curve
o Final constant velocity = 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚
o Why? All enzymes are completely saturated with
substrates, can’t take more substrates

 Meaning: 𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟 𝑜𝑜𝑜𝑜 𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟 = 𝑘𝑘 ∗ [𝑆𝑆]𝟎𝟎 = 𝑘𝑘

𝑲𝑲𝒎𝒎
o At half of 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 the substrate concentration is equal to
the value of 𝑘𝑘𝑚𝑚

Michaelis Menten & Lineweaver-Burk Plot BIOCHEMISTRY: Note #1. 1 of 4

 III) LINEWEAVER-BURK PLOT (ii) Non-competitive inhibition
Apply Michaelis Menten equation to Lineweaver-Burk plot 𝑘𝑘𝑚𝑚 stays → x-intersection doesn’t change
Michaelis Menten equation: 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 decreases → y-intersection moves higher
𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 [𝑆𝑆]
𝑉𝑉0 =
𝑘𝑘𝑚𝑚 + [𝑆𝑆]
Lineweaver-Burk is a linear plot
Linear graph has form:
𝑦𝑦 = 𝑚𝑚𝑚𝑚 + 𝑏𝑏

(1) Transformation into linear equation

Flip it:
1 𝑘𝑘𝑚𝑚 + [𝑆𝑆]
=
𝑉𝑉0 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 [𝑆𝑆]
Split 𝑘𝑘𝑚𝑚 and [𝑆𝑆] over denominator
1 𝑘𝑘𝑚𝑚 [𝑆𝑆]
= +
𝑉𝑉0 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 [𝑆𝑆] 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 [𝑆𝑆]
[𝑆𝑆] cancels out
1 𝑘𝑘𝑚𝑚 1
= +
𝑉𝑉0 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 [𝑆𝑆] 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚
Bring to form: 𝑦𝑦 = 𝑚𝑚 𝑥𝑥 + 𝑏𝑏
1 𝑘𝑘𝑚𝑚 1 1
= ∗ +
𝑉𝑉0 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 [𝑆𝑆] 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚
1 𝑘𝑘𝑚𝑚 1 1 Figure 6: Non-competitive inhibition in Lineweaver-Burk plot
𝑦𝑦 = , 𝑚𝑚 = , 𝑥𝑥 = , 𝑏𝑏 =
𝑉𝑉0 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 [𝑆𝑆] 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚
(i) Uncompetitive inhibition
(2) Plot 𝑘𝑘𝑚𝑚 decreases → x-intersection moves right
X-axis 𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 decreases → y-intersection moves higher
1
o : 1 over substrate concentration
[𝑆𝑆]
1
o Intersection at: −
𝑘𝑘𝑚𝑚

Y-axis
1
o : 1 over velocity
𝑉𝑉0
1
o Intersection at:
𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚

Slope:
𝑘𝑘
o 𝑚𝑚 : km over Vmax
𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚

(i) Competitive inhibition

𝑘𝑘𝑚𝑚 increases → x-intersection moves right
𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚 stays → y-intersection doesn’t change

Figure 7: Uncompetitive inhibition in Lineweaver-Burk plot

Figure 5: Competitive inhibition in Lineweaver-Burk plot

2 of 4 BIOCHEMISTRY: Note #1. Michaelis Menten & Lineweaver-Burk Plot

 IV) APPENDIX

Figure 8: Overview Michaelis curve and Lineweaver-Burk plot

Michaelis Menten & Lineweaver-Burk Plot BIOCHEMISTRY: Note #1. 3 of 4

 V) REVIEW QUESTIONS

1) What is the difference between the Michaelis Menten

(MM) curve and the Lineweaver-Burk (LB) plot?
a) MM is linear while LB is hyperbolic
b) MM is a rectangular hyperbole while LB is linear
c) MM is parabolic while LB is linear
d) There is no difference
2) The plateau part in the MM curve represents:
a) 0 order kinetics
b) 1st order kinetics
c) 2nd order kinetics
d) 3rd order kinetics
3) How will the saturation curve for a competitive
inhibitor look in the LB plot?
a) Intersection with the x-axis is not influenced
b) Intersection with the y-axis is not influenced
c) Intersection with the y-axis is raised
d) Intersection with the y-axis is lowered
4) Which type of inhibition results in a LB plot which is
parallel to the plot for no inhibitor?
a) Competitive inhibition
b) Non-competitive inhibition
c) Uncompetitive inhibition
d) None

CHECK YOUR ANSWERS

4 of 4 BIOCHEMISTRY: Note #1. Michaelis Menten & Lineweaver-Burk Plot