MichaelisMenten kinetics

Concentration

[S]

MichaelisMenten saturation curve for an enzyme reaction

showing the relation between the substrate concentration and reaction rate.

[P]

[E]
[ES]

Time
In biochemistry, MichaelisMenten kinetics is one of
the best-known models of enzyme kinetics. It is named
after German biochemist Leonor Michaelis and Canadian Change in concentrations over time for enzyme E, substrate S,
physician Maud Menten. The model takes the form of complex ES and product P
an equation describing the rate of enzymatic reactions,
by relating reaction rate v to [S] , the concentration of a
where kf , kr , and kcat denote the rate constants,[5] and
substrate S. Its formula is given by
the double arrows between S and ES represent the fact
that enzyme-substrate binding is a reversible process.
v=

d[P ]
Vmax [S]
=
.
dt
KM + [S]

Under certain assumptions such as the enzyme concentration being much less than the substrate concentration
the rate of product formation is given by

Here, Vmax represents the maximum rate achieved by

the system, at maximum (saturating) substrate concentrations. The Michaelis constant KM is the substrate cond[P ]
[S]
[S]
centration at which the reaction rate is half of Vmax .[1] v =
= Vmax
= kcat [E]0
.
dt
KM + [S]
KM + [S]
Biochemical reactions involving a single substrate are often assumed to follow MichaelisMenten kinetics, with- The reaction rate increases with increasing substrate conout regard to the models underlying assumptions.
centration [S] , asymptotically approaching its maximum
rate Vmax , attained when all enzyme is bound to substrate.
It also follows that Vmax = kcat [E]0 , where [E]0 is the
1 Model
initial enzyme concentration. kcat , the turnover number,
is the maximum number of substrate molecules converted
In 1903, French physical chemist Victor Henri found that to product per enzyme molecule per second.
enzyme reactions were initiated by a bond (more gen- The Michaelis constant KM is the substrate concentraerally, a binding interaction) between the enzyme and tion at which the reaction rate is at half-maximum,[1] and
the substrate.[2] His work was taken up by German bio- is an inverse measure of the substrates anity for the
chemist Leonor Michaelis and Canadian physician Maud enzymeas a small KM indicates high anity, meanMenten, who investigated the kinetics of an enzymatic re- ing that the rate will approach Vmax more quickly.[6] The
action mechanism, invertase, that catalyzes the hydrolysis value of KM is dependent on both the enzyme and the
of sucrose into glucose and fructose.[3] In 1913, they pro- substrate, as well as conditions such as temperature and
posed a mathematical model of the reaction.[4] It involves pH.
an enzyme E binding to a substrate S to form a complex
ES, which in turn is converted into a product P and the The model is used in a variety of biochemical situations other than enzyme-substrate interaction, including
enzyme. This may be represented schematically as
antigen-antibody binding, DNA-DNA hybridization, and
protein-protein interaction.[6][7] It can be used to charkf
acterise a generic biochemical reaction, in the same way
kcat
E+P
E + S ES
that the Langmuir equation can be used to model generic
kr
1

DERIVATION

adsorption of biomolecular species.[7] When an empirical

equation of this form is applied to microbial growth, it is
sometimes called a Monod equation.
[E] = [E]0 [ES].
Combining the two expressions above, gives us

Applications

Parameter values vary wildly between enzymes:[8]

kf ([E]0 [ES])[S] = kr [ES].

The constant kcat /KM (catalytic eciency) is a measure Upon simplication, we get
of how an enzyme eciently converts a substrate into
product. Diusion limited enzymes, such as fumarase,
work at the theoretical upper limit of 108 1010 /M*s,
[E]0 [S]
limited by diusion of substrate into the active site.[9]
[ES] =
Kd + [S]
MichaelisMenten kinetics have also been applied to a
variety of spheres outside of biochemical reactions,[5] where Kd = kr /kf is the dissociation constant for the
including alveolar clearance of dusts,[10] the richness enzyme-substrate complex. Hence the velocity v of the
of species pools,[11] clearance of blood alcohol,[12] reaction the rate at which P is formed is[15]
the photosynthesis-irradiance relationship, and bacterial
phage infection.[13]
v=

Derivation

d[P ]
Vmax [S]
=
dt
Kd + [S]

where Vmax = kcat [E]0 is the maximum reaction velocity.

Applying the law of mass action, which states that the rate
of a reaction is proportional to the product of the concentrations of the reactants (i.e.[E][S]), gives a system of 3.2 Quasi-steady-state approximation
four non-linear ordinary dierential equations that dene
An alternative analysis of the system was undertaken
the rate of change of reactants with time t [14]
by British botanist G. E. Briggs and British geneticist J. B. S. Haldane in 1925.[16] They assumed that
the concentration of the intermediate complex does not
d[E]
= kf [E][S] + kr [ES] + kcat [ES]
change on the time-scale of product formation known
dt
as the quasi-steady-state assumption or pseudo-steadyd[S]
= kf [E][S] + kr [ES]
state-hypothesis. Mathematically, this assumption means
dt
kf [E][S] = kr [ES] + kcat [ES] . Combining this relad[ES]
= kf [E][S] kr [ES] kcat [ES]
tionship with the enzyme conservation law, the concendt
tration of complex is[15]
d[P ]
= kcat [ES].
dt
In this mechanism, the enzyme E is a catalyst, which only
[E]0 [S]
[ES] =
facilitates the reaction, so that its total concentration, free
KM + [S]
plus combined, [E] + [ES] = [E]0 is a constant. This
conservation law can also be observed by adding the rst where
and third equations above.[14][15]

3.1

Equilibrium approximation

KM =

kr + kcat
kf

In their original analysis, Michaelis and Menten assumed is known as the Michaelis constant, where kr , kcat , and
that the substrate is in instantaneous chemical equilibrium kf are, respectively, the constants for substrate unbinding,
with the complex, which implies[4][15]
conversion to product, and binding to the enzyme. Hence
the velocity v of the reaction is[15]
kf [E][S] = kr [ES].
From the enzyme conservation law, we obtain[15]

v=

d[P ]
Vmax [S]
=
.
dt
KM + [S]

3.3

Assumptions and limitations

The rst step in the derivation applies the law of mass action, which is reliant on free diusion. However, in the
environment of a living cell where there is a high concentration of proteins, the cytoplasm often behaves more
like a gel than a liquid, limiting molecular movements and
altering reaction rates.[17] Whilst the law of mass action
can be valid in heterogeneous environments,[18] it is more
appropriate to model the cytoplasm as a fractal, in order
to capture its limited-mobility kinetics.[19]

[S] [P ].
This is true under standard in vitro assay conditions, and
is true for many in vivo biological reactions, particularly
where the product is continually removed by a subsequent
reaction.

2. The energy released in the reaction is very

large, that is
The resulting reaction rates predicted by the two approaches are similar, with the only dierence being that
the equilibrium approximation denes the constant as Kd
, whilst the quasi-steady-state approximation uses KM .
G 0.
However, each approach is founded upon a dierent assumption. The MichaelisMenten equilibrium analysis is
valid if the substrate reaches equilibrium on a much faster In situations where neither of these two conditions hold
time-scale than the product is formed or, more precisely, (that is, the reaction is low energy and a substantial pool of
that [15]
product(s) exists), the MichaelisMenten equation breaks
down, and more complex modelling approaches explicitly taking the forward and reverse reactions into account
kcat
must be taken to understand the enzyme biology.
d =
1.
kr
By contrast, the BriggsHaldane quasi-steady-state analysis is valid if [14][20]

m =

[E]0
1.
[S]0 + KM

Thus it holds if the enzyme concentration is much less

than the substrate concentration. Even if this is not satised, the approximation is valid if KM is large.
In both the MichaelisMenten and BriggsHaldane analyses, the quality of the approximation improves as decreases. However, in model building, MichaelisMenten
kinetics are often invoked without regard to the underlying assumptions.[15]
It is also important to remember that, while irreversibility
is a necessary simplication in order to yield a tractable
analytic solution, in the general case product formation
is not in fact irreversible. The enzyme reaction is more
correctly described as

kf1

kf2

kr1

kr2

E + S ES E + P.

4 Determination of constants
The typical method for determining the constants Vmax
and KM involves running a series of enzyme assays at
varying substrate concentrations [S] , and measuring the
initial reaction rate v0 . 'Initial' here is taken to mean
that the reaction rate is measured after a relatively short
time period, during which it is assumed that the enzymesubstrate complex has formed, but that the substrate
concentration held approximately constant, and so the
equilibrium or quasi-steady-state approximation remain
valid.[20] By plotting reaction rate against concentration,
and using nonlinear regression of the MichaelisMenten
equation, the parameters may be obtained.[21]
Before computing facilities to perform nonlinear regression became available, graphical methods involving linearisation of the equation were used. A number of these
were proposed, including the EadieHofstee diagram,
HanesWoolf plot and LineweaverBurk plot; of these,
the HanesWoolf plot is the most accurate.[21] However,
while useful for visualization, all three methods distort
the error structure of the data and are inferior to nonlinear regression.[22] Nonetheless, their use can still be found
in modern literature.[23]

In general, the assumption of irreversibility is a good one In 1997 Santiago Schnell and Claudio Mendoza suggested
a closed form solution for the time course kinetics analin situations where one of the below is true:
ysis of the MichaelisMenten kinetics based on the solution of the Lambert W function.[24] Namely:
1. The concentration of substrate(s) is very
much larger than the concentration of products:

[S]
= W (F (t))
KM

where W is the Lambert W function and

(
)
[S]0
[S]0
Vmax
F (t) =
exp

t .
KM
KM
KM

REFERENCES

[5] Chen, W.W.; Neipel, M.; Sorger, P.K. (2010). Classic

and contemporary approaches to modeling biochemical reactions.
Genes Dev 24 (17): 18611875.
doi:10.1101/gad.1945410. PMC 2932968. PMID
20810646.

The above equation has been used to estimate Vmax and

KM from time course data.[25][26]

[6] Lehninger, A.L.; Nelson, D.L.; Cox, M.M. (2005).

Lehninger principles of biochemistry. New York: W.H.
Freeman. ISBN 978-0-7167-4339-2.

[7] Chakraborty, S. (23 Dec 2009). Microuidics and Microfabrication (1 ed.). Springer. ISBN 978-1-4419-1542-9.

Role of substrate unbinding

[8] Mathews, C.K.; van Holde, K.E.; Ahern, K.G. (10 Dec

The Michaelis-Menten equation has been used to pre1999). Biochemistry (3 ed.). Prentice Hall. ISBN 978-0dict the rate of product formation in enzymatic reactions
8053-3066-3.
for more than a century. Specically, it states that the
rate of an enzymatic reaction will increase as substrate [9] Stroppolo, M.E.; Falconi, M.; Caccuri, A.M.; Desideri,
concentration increases, and that increased unbinding of
A. (Sep 2001). Superecient enzymes. Cell Mol Life
enzyme-substrate complexes will decrease the reaction
Sci 58 (10): 145160. doi:10.1007/PL00000788. PMID
11693526.
rate. While the rst prediction is well established, the second has never been tested experimentally. To determine
whether an increased rate of unbinding does in fact de- [10] Yu, R.C.; Rappaport, S.M. (1997). A lung retention model based on MichaelisMenten-like kinetcrease the reaction rate, Shlomi Reuveni et al. mathematics.
Environ Health Perspect 105 (5): 496503.
ically analyzed the eect of enzyme-substrate unbinddoi:10.1289/ehp.97105496. PMC 1469867. PMID
ing on enzymatic reactions at the single-molecule level.
9222134.
According to the study, unbinding of an enzyme from a
substrate can reduce the rate of product formation under [11] Keating, K.A.; Quinn, J.F. (1998). Estimating species
some conditions, but may also have the opposite eect.
richness: the MichaelisMenten model revisited. Oikos
As substrate concentrations increase, a tipping point can
81 (2): 411416. doi:10.2307/3547060. JSTOR
3547060.
be reached where an increase in the unbinding rate results
in an increase, rather than a decrease, of the reaction rate.
The results indicate that enzymatic reactions can behave [12] Jones, A.W. (2010). Evidence-based survey of the elimination rates of ethanol from blood with applications in
in ways that violate the classical Michaelis-Menten equaforensic casework. Forensic Sci Int 200 (13): 120.
tion, and that the role of unbinding in enzymatic catalysis
doi:10.1016/j.forsciint.2010.02.021. PMID 20304569.
[27]
still remains to be determined experimentally.

See also

[13] Abedon, S.T. (2009). Kinetics of phage-mediated biocontrol of bacteria. Foodborne Pathog Dis 6 (7): 80715.
doi:10.1089/fpd.2008.0242. PMID 19459758.

Enzyme kinetics

[14] Murray, J.D. (2002). Mathematical Biology: I. An Introduction (3 ed.). Springer. ISBN 978-0-387-95223-9.

LineweaverBurk plot
Reaction progress kinetic analysis
Steady state (chemistry)

References

[1] http://www.worthington-biochem.com/introbiochem/
substrateconc.html
[2] Henri, Victor (1903). Lois Gnrales de lAction des Diastases. Paris: Hermann. Google books (US only)
[3] Victor Henri. Whonamedit?. Retrieved 24 May 2011.
[4] Michaelis, L.; Menten, M.L. (1913). Die Kinetik der
Invertinwirkung. Biochem Z 49: 333369 (recent translation, and an older partial translation)

[15] Keener, J.; Sneyd, J. (2008). Mathematical Physiology: I:

Cellular Physiology (2 ed.). Springer. ISBN 978-0-38775846-6.
[16] Briggs, G.E.; Haldane, J.B.S. (1925). A note on the kinematics of enzyme action. Biochem J 19 (2): 338339.
PMC 1259181. PMID 16743508.
[17] Zhou, H.X.; Rivas, G.; Minton, A.P. (2008).
Macromolecular crowding and connement: biochemical, biophysical, and potential physiological
consequences. Annu Rev Biophys 37 (1): 37597.
doi:10.1146/annurev.biophys.37.032807.125817. PMC
2826134. PMID 18573087.
[18] Grima, R.; Schnell, S. (Oct 2006).
A systematic investigation of the rate laws valid in intracellular environments. Biophys Chem 124 (1): 110.
doi:10.1016/j.bpc.2006.04.019. PMID 16781049.

[19] Schnell, S.; Turner, T.E. (2004). Reaction kinetics in

intracellular environments with macromolecular crowding: simulations and rate laws. Prog Biophys Mol Biol 85
(23): 23560. doi:10.1016/j.pbiomolbio.2004.01.012.
PMID 15142746.
[20] Segel, L.A.; Slemrod, M. (1989). The quasi-steady-state
assumption: A case study in perturbation. Thermochim
Acta 31 (3): 446477. doi:10.1137/1031091.
[21] Leskovac, V. (2003). Comprehensive enzyme kinetics.
New York: Kluwer Academic/Plenum Pub. ISBN 9780-306-46712-7.
[22] Greco, W.R.; Hakala, M.T. (1979). Evaluation of methods for estimating the dissociation constant of tight binding enzyme inhibitors,. J Biol Chem 254 (23): 12104
12109. PMID 500698.
[23] Hayakawa, K.; Guo, L.; Terentyeva, E.A.; Li, X.K.;
Kimura, H.; Hirano, M.; Yoshikawa, K.; Nagamine,
T.; et al. (2006). Determination of specic activities and kinetic constants of biotinidase and lipoamidase
in LEW rat and Lactobacillus casei (Shirota)". J Chromatogr B Analyt Technol Biomed Life Sci 844 (2): 24050.
doi:10.1016/j.jchromb.2006.07.006. PMID 16876490.
[24] Schnell, S.; Mendoza, C. (1997). A closed form
solution for time-dependent enzyme kinetics.
Journal of Theoretical Biology 187 (2): 207212.
doi:10.1006/jtbi.1997.0425.
[25] Goudar, C. T.; Sonnad, J. R.; Duggleby, R. G. (1999).
Parameter estimation using a direct solution of the integrated MichaelisMenten equation. Biochimica et
Biophysica Acta Protein Structure and Molecular Enzymology 1429 (2): 377383. doi:10.1016/s01674838(98)00247-7. PMID 9989222.
[26] Goudar, C. T.; Harris, S. K.; McInerney, M. J.; Suita, J. M. (2004). Progress curve analysis for enzyme and microbial kinetic reactions using explicit solutions based on the Lambert W function. Journal of Microbiological Methods 59 (3): 317326.
doi:10.1016/j.mimet.2004.06.013. PMID 15488275.
[27] Reuveni, Shlomi;
Urbakh, Michael;
Klafter,
Joseph (2014).
Role of Substrate Unbinding in
Michaelis-Menten Enzymatic Reactions.
Proceedings of the National Academy of Sciences 111
(12): 43914396. Bibcode:2014PNAS..111.4391R.
doi:10.1073/pnas.1318122111.

Further reading
Biochemistry/Catalysis at Wikibooks

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