This document discusses different plots that can be used to determine the kinetic parameters Km and Vmax from enzyme kinetics experiments. It explains that Vmax is the maximum reaction rate when the enzyme is fully saturated, while Km is the substrate concentration at half Vmax. It then describes the Michaelis-Menten equation and how it can be used to model enzyme-catalyzed reactions. Several different types of plots are introduced, including Lineweaver-Burk, Eadie-Hofstee, Hanes, and Eisenthal-Cornish-Bowden plots, which transform the Michaelis-Menten equation to allow Km and Vmax to be determined graphically from kinetic data.

1. Different Plots for
Determination of Km and
Vmax
SUBMITTED BY: Amir sohil
SUBMITTED TO: Bharat Bhusan Sir

2. Vmax
Vmax is the reaction rate when the
enzyme is fully saturated by substrate,
indicating that all the binding sites are
being constantly reoccupied.
Km
Km is a value of substrate concentration at
half maximal velocity

4. Michaelis-Menten Equation
Michaelis –Mentum kinetics is one of the
simplest and best known model of enzyme
kinetics.
This model explain how an enzyme can
cause kinetic rate enhancement of a
reaction and why the rate of reaction
depends on the concentration of enzyme
present.

6. According to this model the enzyme
reversibly bind with substrate to form an ES
complex that subsequently yield product.

7. The mechanism of enzyme catalyzed
reactions is studied by making kinetic
measurements on enzyme substrate
reaction system
The simple kinetic model os obtained,
when plotting the rate of catalysis,V
(reaction velocity),V/S the substrate
concentration [S]

8. From this equation, the reaction velocity,V
can be expressed as
.

9. Lineweaver-Burk Plot
 The Lineweaver-Burk plot is a
representation of the Lineweaver-Burk
Equation of Enzyme kinetics,described by
Lineweaver and Dean Burk in 1934.

11. 
Taking the reciprocal of this equation
gives

12. This plot was widely used to determine Km
and Vmax, before the wide availability of
powerful computers and non-linear
regression Software.

13. Eadie-Hofstee and Hanes Plot
The Lineweaver-Burk plot has been criticized
on several grounds
Firstly the extrapolation across the 1/Vo axis
determine _1/Km sometimes reaches the
of graph paper before reaching 1/[S] axis
Secondly, it is said to give undue weight to
measurements made at low concentration.

14. Eadie-Hofstee Plot

15. The Eadie-Hofstee plot takes as its
starting point the Lineweaver-Burk
equation.Both sides of the equation are
multiplied by the factor Vo V max :

16. The Hanes plot (devised by Charles
Hanes) similarly starts with the Lineweaver-
Burk equationwhich in this instance is
multiplied throughout by [S]

18. Eisenthal and Cornis Bowden Plot
In view of the difficulties in obtaining reliable
estimates of Km and V max , even with
the aid of statistical analysis, from the plots
discussed above, Robert Eisenthal and
Athel Comish-Bowden (1974) suggested a
different approach, though one still based
on the Michaelis-Menten equation (7.12). The
reciprocal form of this equation, at
constant [E0], gives

20. 
Therefore, at constant v0 and [S0], a plot
of V max against Km is linear.

21. References
Palmer Trevor,Bonnner Philip
Enzymes second edition
Gupta N.S Biochemistry