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Soil Biology & Biochemistry 57 (2013) 946e949

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Soil Biology & Biochemistry

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Short communication

A note on the reverse MichaeliseMenten kineticsq

Gangsheng Wang*, Wilfred M. Post
Climate Change Science Institute and Environmental Sciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831-6301, USA

a r t i c l e i n f o

a b s t r a c t

Article history:
Received 1 June 2012
Received in revised form
27 August 2012
Accepted 29 August 2012
Available online 13 September 2012

We theoretically derived a general equation describing the enzyme kinetics that could be further
simplied to the typical MichaeliseMenten (MeM) kinetics or the reverse MeM equation (RMeM)
under the condition of S z S1 >> E0 or S1 << E0, respectively, where E0 and S1 ( S ES) are the
concentrations of total enzyme and substrate including free substrate (S) and enzymeesubstrate
complex (ES). We showed that the related Schimel and Weintraub RMeM equation (RMeMeSW) can
be derived from the Langmuir adsorption isotherm theory with S >> E0. Both the MeM and the RM-MSW are appropriate to eld soil conditions with S >> E0 given different values of specic reaction rate
(k3) and half-saturation constant (Ks). In contrast to MeM and RMeMeSW models, the RMeM model is
not applicable to eld conditions because of its limited application to one substrate with a simple
enzyme system. However, we demonstrate that the best formulation for the process of enzyme-mediated
decomposition may vary depending on whether the process is limited by substrate or enzyme
Published by Elsevier Ltd.

Enzyme kinetics
Langmuir adsorption isotherm
MichaeliseMenten kinetics
Reverse MichaeliseMenten kinetics

1. Introduction
Soil enzymes have been explicitly included in the modeling of
soil organic matter (SOM) dynamics (Lawrence et al., 2009). In
addition to the MichaeliseMenten (MeM) kinetics (Allison et al.,
2010; Wang et al., in press), the reverse MichaeliseMenten model
(RMeMeSW) proposed by Schimel and Weintraub (2003) has been
used to describe soil enzyme kinetics. The RMeMeSW kinetics
functionally reverses the dependency of the MeM equation on
substrate availability (Schimel and Weintraub, 2003). The size of
the SOM pool in the RMeMeSW is assumed to be sufciently large
so that the amount of enzyme, rather than substrate, is the ratelimiting factor for SOM decomposition (Lawrence et al., 2009;
Moorhead and Sinsabaugh, 2006; Schimel and Weintraub, 2003).
The typical MeM and the RMeMeSW can be expressed by Eqs. (1)
and (2), respectively

q This manuscript has been authored by UT-Battelle, LLC, under Contract No. DEAC05-00OR22725 with the U.S. Department of Energy. The United States Government retains and the publisher, by accepting the article for publication, acknowledges that the United States Government retains a non-exclusive, paid-up,
irrevocable, world-wide license to publish or reproduce the published form of this
manuscript, or allow others to do so, for United States Government purposes.
* Corresponding author. Bldg 2040, Room E272, MS-6301, Oak Ridge National
Laboratory, Oak Ridge, TN 37831-6301, USA.
E-mail address: (G. Wang).
0038-0717/$ e see front matter Published by Elsevier Ltd.


k3;M $E0 $S
Ks;M S



k3;R $E0 $S
Ks;R E0


where the subscript M and R in v, k3, and Ks denote the MeM and
RM-M-SW kinetics, respectively; E0 and S represent the concentration of total enzyme and free substrate, respectively; v is the
SOM decomposition rate; k3 denotes the specic rate constant and
k3,M in Eq. (1) is equivalent to the turnover number (kcat) (Johnson
and Goody, 2011); Ks denotes the half-saturation constant. Mathematically, the RMeMeSW (Eq. (2)) uses the enzyme pool instead
of the substrate pool in the denominator of the typical MeM
equation (Eq. (1)).
This RMeMeSW formulation is different from the reverse
MichaeliseMenten equation described in Bowie et al. (1985). The
rate-limitation term of Bowie et al. (1985) has the form of Ks/
(Ks S) rather than S/(Ks S) as in the typical MeM equation. As
stated by Schimel and Weintraub (2003), the adoption of the
nonlinear RMeMeSW kinetics makes it possible to construct
a stable system by providing a saturating response, while the rstorder enzyme kinetics could result in an unstable microbial
biomass pool in their C-only model because of a lack of nonlinear
kinetics. Actually, the rst-order kinetics used by Schimel and
Weintraub (2003) is a simplication of the MeM equation since

G. Wang, W.M. Post / Soil Biology & Biochemistry 57 (2013) 946e949

the substrate pool (S) is assumed to be large and relatively

invariant, i.e., S/(Ks,M S) in Eq. (1) becomes constant.
The MeM kinetics is theoretically derivable for a simple enzyme
with only one active site and a single substrate (Briggs and Haldane,
1925; Johnson and Goody, 2011). The RMeMeSW model was
introduced as an empirical equation rather than one derived on
a theoretical basis (Schimel and Weintraub, 2003). In this paper, we
provided theoretical derivations of the enzyme kinetics models and
elaborated under what conditions these models are valid.

free substrate (S) and the enzyme-bound substrate (ES), while

Robson and Garnier (1986) referred to the total substrate (denoted
by S0) in the left side of Eq. (11) since the enzyme concentration was
considered to be signicant compared to substrate concentration
and the product concentration (P) was negligible in their case
(Easterby, 1981; Leskovac, 2003). We can replace v with Vmax/2 and
solve for S1 in Eq. (11) to provide an estimate of the apparent halfsaturation concentration of S1

2. Derivation of the reverse MichaeliseMenten equation





where S and P are the concentration of substrate and product,

respectively, ES is the intermediate enzymeesubstrate complex
which reacts to form the product P, and results in the release of the
enzymes (E). The rate constants k1 and k3 denote the forward
reactions, and the rate constant k2 refers to a reverse reaction
(Johnson and Goody, 2011).
One of the important assumptions in the MeM kinetics is that
the concentration of intermediate species (ES) is quasi-steady, i.e.,
the rate of production of ES equals the rate of degradation of ES
(Briggs and Haldane, 1925). This assumption yields

E$S Ks $ES


where Ks (k2 k3)/k1 is the half-saturation constant, i.e., the

substrate concentration at which the reaction rate v Vmax/2; and
Vmax is the maximum reaction rate.
The reaction (production) rate is proportional to the concentration of ES:

v dP=dt k3 $ES


The maximum reaction rate (Vmax) can be reached under the

condition that all enzyme (E0) is converted to ES:

Vmax k3 $E0


where E0 consists of two forms, i.e., the free enzyme (E) and the
enzymeesubstrate complex (ES):


where A Ks E0 S1 and B 4E0$S1. Since A2 e
B (Ks)2 2$Ks$(E0 S1) (E0  S1)2 > 0, B/A2 < 1, i.e.,

4E0 $S1 =Ks E0 S1 2 < 1


Based on the Taylor series expansion (Fowler and Robson, 1998;

Moller et al., 1997) and Eq. (14), the truncated rst-order approximation to the square root in Eq. (13) can be expressed as

A2  BzA  B=2A


It follows that


A2  BzB=2A 2E0 $S1 =Ks E0 S1


Substituting Eq. (15b) into Eq. (13) with Vmax k3$E0 from Eq.
(6) yields

Vmax $S1
k3 $E0 $S1

Ks E0 S1
Ks E0 S1


Equation (16) shows another viewpoint that Ksapp Ks E0 by

replacing v with Vmax/2 and solving S1. Equation (16) may be further
simplied into:


k $E $S
vz 3 0 1 ; if S1 << E0
Ks E0




Replacing ES in Eq. (10) with Eq. (8) and rearranging Eq. (10)

S1 Ks $v=Vmax  v E0 $v=Vmax

v Vmax =2E0 $ Ks E0 S1  Ks E0 S1 2 4E0 $S1

Vmax =2E0 $ A  A2  B


With E E0  ES and S S1  ES from Eqs. (7) and (9),

respectively, Eq. (4) becomes

E0  ES$S1  ES Ks $ES

where Ks
is called the apparent or effective Ks by Robson and
Garnier (1986). The Ksapp is dependent on enzyme concentration
and larger than the intrinsic half-saturation constant (Ks).
The solution to Eq. (11) for v is

k $E $S
k $E $S
vz 3 0 1 z 3 0 ; if S1 >>E0
Ks S
Ks S1

We dene S1 as the sum of the free substrate and the enzymebound substrate:




Combining Eqs. (5) and (6) gives

ES E0 $v=Vmax

S1 jvVmax =2 Ks E0 =2

The enzyme-catalyzed reaction is usually illustrated as follows

(Johnson and Goody, 2011; Leskovac, 2003):

S E % ES / E P



Eq. (11) is similar to the equation (4.11) described by Robson and

Garnier (1986). However, S1 in our formula denotes the sum of the

Equations (16) and (17a,b) indicate that (i) under the condition
of S1 >> E0, the reaction rate can be expressed by the typical MeM
kinetics since S z S1 (Eq. (17a) equivalent to Eq. (1)); (ii) in the case
of S1 << E0, the reaction rate (Eq. (17b)) has a similar formulation to
the RMeMeSW equation (Eq. (2)); for convenience, we named Eq.
(17b) as the RMeM equation to distinguish it from the RMeMeSW;
and (iii) when the enzyme concentration is comparable to the
substrate concentration, both S1 and E0 should be included in the
denominator as indicated by Eq. (16).
The above derivation is appropriate for a simple enzyme and
a single substrate. However, there are many enzymes competing for
the binding sites on a broad range of insoluble substrates in soils
(Schimel and Weintraub, 2003). The limiting factor is the availability of effective binding sites on solid substrates (Sinsabaugh and


G. Wang, W.M. Post / Soil Biology & Biochemistry 57 (2013) 946e949

Table 1
Comparisons of three models: MichaeliseMenten (MeM), reverse MeM (RMeM) and Schimel and Weintraub RMeM (RMeMeSW).
Theoretical basis
More sensitive to enzyme
than to substrate


k3;M $E0 $S
Ks;M S
Enzyme catalysis as a chain reaction
S z S1 >> E0
Simple enzyme/single substrate;
many enzymes/complex substrates


S1 << E0

k3;R $E0 $S
Ks;R E0
Langmuir adsorption isotherm theory
S >> E0

Simple enzyme/single substrate

Many enzymes/complex substrates


k3;M $E0 $S1

Ks;M E0


Mathematical equations for different models, E0 and S represent the concentration of total enzyme and free substrate, respectively; S1 S ES, where ES is the
concentration of enzymeesubstrate complex; v is the reaction rate; k3 denotes the maximum specic rate constant; Ks denotes the half-saturation constant; and the subscript
in v, k3, and Ks denote different models.

Osgood, 2007). As indicated by Schimel and Weintraub (2003), we

can use a formulation analogous to the Langmuir equation (Sohn
and Kim, 2005) to estimate the fraction of binding sites with the
enzyme as the adsorbate:

1 KBA $E0


where q is the fraction of binding sites (the ratio of the number of

binding sites occupied to the number of available sites) and KBA is
the binding afnity. If we assume that (i) the maximum binding
capacity is proportional to the concentration of substrate (S) and (ii)
the decomposition rate (vR) is proportional to the binding amount
of enzyme, vR can be estimated as

a$E0 $S
vR a$q$S a$
1=KBA E0
1 KBA $E0


where a is an integrated proportional coefcient taking into

account the aforementioned two assumptions. Eq. (19) is equivalent to Eq. (2) if a and 1/KBA are replaced by k3,R and Ks,R, respectively. Therefore, the RMeMeSW can be derived from the Langmuir
adsorption isotherm theory.
3. Model comparisons
A summary of the model comparisons is shown in Table 1. The
MeM and the RMeM are theoretically derived for a simple enzyme
and a single substrate under opposite conditions in terms of the
concentrations of enzyme and substrate. When the system is
substrate limited (S1 << E0), the RMeM is valid. The MeM equation
does not literally apply to complex systems such as SOM
(Sinsabaugh and Osgood, 2007). The derived k3 and Ks by tting the
v w S relationship from laboratory experiments with a simple
enzyme and a single substrate cannot be directly used for eld soil
conditions (Wang et al., 2012). In ecosystems, the parameters (k3
and Ks) of the MeM model are often not independent and are
functions of the methods and conditions of measurement
(Sinsabaugh and Osgood, 2007). The RMeMeSW may be derived
from the Langmuir adsorption isotherm theory, which is theoretically based on the statistical rate theory of interfacial transport
(Rudzinski and Panczyk, 2000; Ward et al., 1982). While RMeM is
supercially similar to the RMeMeSW, the RMeM is actually
different in principle from the RMeMeSW since the latter is
applicable under enzyme-limited conditions (S >> E0). In addition,
as shown from our derivation, most of the substrate is bound to the
enzyme and S1 in the RMeM (Eq. (17b)) may not be changeable
with S when S1 << E0.
Is the reaction rate more sensitive to changes in enzyme
concentration than to changes in substrate concentration when the
system is enzyme-limited (S >> E0)? To answer this question we

conducted a sensitivity analysis of both the MeM and the RM-MSW equations. Following the approach of Wang and Post (2012),
the sensitivities of the reaction rate (vM) in the MeM kinetics to the
changes in the concentrations of enzyme (E0) and substrate (S) are

vvM vM 

 k3;M $S$E0
Ks;M S vM



vvM vM 
k3;M S$E0
 Ks;M $

Ks;M S Ks;M S vM


respectively. Similarly the sensitivities for the RMeMeSW equation

(vR) to changes in E0 and S are


 vvR vR 

Ks;R E0 Ks;R E0 vR


vvR vR 

Ks;R E0 vR


Eq. (20a) and (20b) indicate that, with the same S and E0 values,
the reaction rate (vM) of the MeM model is more sensitive to the
changes in enzyme concentration (E0) than to the changes in S since
Ks,M/(Ks,M S) < 1 regardless of the concentrations of S and E0. On
the contrary, vR of the RMeMeSW model is more sensitive to the
changes in S than to the changes in E0 because of Ks,R/(Ks,R E0) < 1.
Similarly, the RMeM model is more sensitive to substrate than to
enzyme. The above analysis indicates that the relative sensitivities
of the three models are independent of the relative sizes of S and E0.
It also implies that the relative sensitivity of a response variable to
a dependent variable is determined by the response mechanism.
Both the MeM and RMeMeSW have been shown to be useful in
simulating SOM dynamics (Allison et al., 2010; Schimel and
Weintraub, 2003) with S >> E0 and appropriate values of k3 and
Ks. It is possible for the reaction rates for both formulations to be the
same. This occurs when k3,M/(Ks,M S) k3,R/(Ks,R E0) (see Eqs. (1)
and (2)). However, the sensitivities of the reaction rates to enzyme
or substrate availabilities are quite different. Under the condition of
k3,M/(Ks,M S) k3,R/(Ks,R E0), when the MeM is used, the reaction
rate is more sensitive to enzyme concentration (E0) than when the
RMeMeSW is used since Ks,R/(Ks,R E0) < 1 resulting in Eq.
(20a) > Eq. (21a).

4. Conclusion
Both the MeM and the RMeMeSW models are ecologicallyapplicable. The MeM kinetics is theoretically derivable for
a simple enzyme with only one active site and a single substrate

G. Wang, W.M. Post / Soil Biology & Biochemistry 57 (2013) 946e949

(Briggs and Haldane, 1925; Johnson and Goody, 2011). When the
MeM model is used to model SOM dynamics with many enzymes
and complex substrates, it is an empirical analogy rather than
a theoretical model. The RMeMeSW can be derived from the
Langmuir adsorption isotherm theory. The derivation of the RMeM
equation requires assumptions that are contrary to those required
for RMeMeSW, which limits the applicability of the RMeM to eld
soil conditions. We demonstrate that the best formulation for the
process of enzyme-mediated decomposition may vary depending
on whether the process is limited by substrate or enzyme
Research sponsored by the Laboratory Directed Research and
Development Program of Oak Ridge National Laboratory, managed
by UT-Battelle, LLC, for the U.S. Department of Energy under
contract No. DE-AC05-00OR22725. The authors thank Dr. Xiaojuan
Yang for her technical comments. The authors are also grateful for
the constructive comments and suggestions from the three anonymous reviewers.
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