The Monod Equation: A Revisit and a

Generalization to Product Inhibition Situations

OCTAVE LEVENSPI EL, Cliernical Engineering Depnrtment

Oregon State Uni\.ersity, Coriwllis, Oregon 97331

Summary
This paper shows how to treat the substrate-limiting Monod equation in a straight-
forward manner for different types of fermentors (plug-flow, batch, and mixed-flow)
using the general language of chemical reaction engineering. Straight-line plots are
developed for directly finding the kinetic constants of the equation, and an example
using Monod's original data illustrates the procedure. The Monod equation is then
generalized to account for the effects of both substrate and inhibitory toxic wastes.
Finally. for pure product inhibition performance. expressions are derived for various
reactor types, and correlation graphs are developed for finding the kinetic constants
of the reaction. An example from the recent literature shows that this equation form
fits the data extremely well.

I. INTRODUCTION

Although the growth of microorganisms is an unusually complex

phenomenon, it is often possible to represent this growth by rela-
tively simple laws. In particular, Monod's pioneering pro-
posed a very simple equation form whenever a single essential
nutritional requirement is the growth-limiting factor. and where the
presence of toxic metabolic products plays no role. In the general
language of chemical reactors Monod's equation is expressed as
(sub y e ) +cells ,(proFct) + (moreccells)

and
rc = kC,Cc/(CA + C,) (1)
where r c is the rate of production of cells: C , is the substrate
concentration; C c is the cell concentration; C w is the Monod con-
stant, the substrate concentration where the rate is one-half the
maximum.
This paper proposes a simple generalization of the Monod equa-

Biotechnology and Bioengineering, Vol. XXII, Pp. 1671- 1687 (1980)

@ 1980 John Wiley & Sons, Inc. 00063592/80/0022-1671$01.70
I672 LEV EN SPI EL

tion to account for the influence of inhibitory product R, as follows:

I'r. = k(l - + C,)]
C,/CR*)n.[CACC/(CA (2a)
or
rC = k < , , , , [ C A C C / (+C A
C,)] with k,,,,, = k ( l - C,/C,")" (2b)
where C,* is the limiting concentration of inhibitory product above
which cells cease to multiply. At the extreme where C R e CR*,
inhibition by product does not slow the rate and so
k ( 1 - CR/CR*)n + k or kobs + k
in which case, eq. (2) reduces to the original inhibition-free sub-
strate-limiting Monod expression of eq. ( I ) .
At the other extreme where C A %- Cw,the depletion of substrate
does not slow the rate,
CA/(CA + CAW)+=1
and eq. (2) reduces to substrate-independent and purely inhibitory
kinetics, as follows:
rc = k ( 1 - C,/CR*pCc (3 )
Section I1 shows how to treat the substrate-limiting Monod
expression of eq. ( 1 ) in a straightforward manner for various reactor
types: batch, plug-flow and mixed-flow [or continuous stirred tank
reactor (CSTR), chemostat, turbidostat, etc.]. Section I11 does the
same for the pure product inhibition expression of eq. (3), while
Sec. IV considers situations where the generalized Monod expres-
sion of eq. (2) must be used. There, both depletion of nutrient and
the accumulation of inhibitory product influence the progress of
reaction.
Two examples, one using Monod's original data, the other re-
quiring the use of the generalized expression, show how to find the
kinetic constants simply and directly from reported data.
We make one simplifying assumption in this treatment, to the
effect that every unit of substrate consumed will produce the same
amount of cells, and of product R , at all conditions. Thus
 THE MONOD EQUATION 1673

where [AIC],[AIR],and [CIR]are called the yields. Thus we assume

throughout that the yields stay unchanged for any system. This is
often a reasonable assumption, as was shown by Monod,’ Bazua
and Wilke.7 and others.

11. SUBSTRATE-LIMITING MONOD KINETICS

Here we develop the performance equations for the Monod ki-

netics of eq. ( 1 ) occurring in various ideal reactor types. Since much
of this is straightforward we -will be brief.

Batch or Plug-Flow Reactor

For a liquid-phase system the basic performance equation, given
in any reactor design book (e.g., see Ref. 3), is as follows:

Inserting eq. ( I ) , converting to one variable with eq. (4),and then

integrating gives the changing concentration of materials as

This expression gives an S-shaped concentration-time curve as

shown in Figure 1.
It is difficult to evaluate the kinetic constants from the integrated

-
‘run out of food

d e v i a t e s from exponential
at if both CA a n d Cc c h a n g e
start appreciably during t h e run
$
cco
0
- tb Or T p

Fig. I . Concentration-time curve for substrate-limiting Monod kinetics in a batch

or plug-flow reactor.
1674 LEVENSPIEL

expression of eq. (6), hence it is preferable to use the differential

rate form of eq. ( I ) directly. For this take small time intervals At,
and for each evaluate C,,, C(., and Fc. = ACJ A t . Then rearranging
eq. ( I ) gives

C C / G = l/k + ( C . d W / C A ) (7)
The plot of Figure 2 then shows how to evaluate the Monod con-
stants from either batch or plug-flow experiments. They are given
by the slope and intercept of the best straight line through the data.
Extension of these equations to plug flow with recycle and to
optimum operations are given in Ref. 3.

Example of Substrate-Limiting Inhibition-Free Kinetics

Monod (Ref. I , p. 74) presents the following data on the growth
of Escherichia coli cultures (cells C ) in a lactose medium (substrate
A ) . Find a rate equation of the Monod type to represent the kinetics
of this growth.

Solution
Make the tabulation given in Table I. Plotting the last two columns
as in Figure 3, we find

rC = 0.72 CACC/(CA + 26.3) (84

The constants of this expression differ substantially from the values
reported by Monod. His final rate expression, found by fitting a

- -I 0
CM

Fig. 2. Graphical procedure for finding the Monod constants of eq. ( I ) from
batch or plug-flow data. Data points are included for illustration only.
 THE MONOD EQUATION 1675

I , t I 1 I ,
0 01 02 03 04 05

Fig. 3. Plot of C J F , . vs. liC., for the batch reactor data of Table I.

curve to the four sets of rr./Cpvs. c,,data of Table I is given as

rc = 1.2 CACc/(CA + 20) (8b)
The striking difference in eqs. (8a) and (8b) shows that the method
of analyzing data can greatly influence the results obtained.

Mixed-Flow Reactor
The basic performance equation for steady-state mixed flow and
any kinetics is given by (see Ref. 3)
- V
- Ci,emt - Ci,enter
TI,, = - - (9)
V T i , at vxit wnclition

where i is any reactant or product.

For a feed containing substrate A but no cells C , insertion of eq.
( I ) into eq. (9), and converting to one variable with eq. (4) gives, in
terms of A or of C

This expression was developed independently by Monod4 and by

Novick and S ~ i l a r d . ~
The features of this remarkable equation are shown in Figure 4,
which is a more convenient version of the graph first presented by
Herbert? Note that all quantities such as washout, maximum pro-
 TABLE 1
Monod's Reported Batch Reactor Data (Run I ) , Reanalyzed
Given Calculated
r
Interval At C .I C, c, r, = AC,lAt c (.IF(. llC* rn
<
I 0.54 137 15.5-23.0 19.25 13.89 I .39 0.0073 rn
21
2 0.36 1 I4 23.0-30.0 26.5 19.44 I .36 0.0088 vl
3 0.33 90 30.0- 38.8 34.4 26.67 1.29 0.01 1 I
a
-
m
4 0.35 43 38.8-48.5 43.65 27.1 I I .58 0.0233 r
5 0.37 29 48.5- 58.3 53.4 26.49 2.02 0.0345
6 0.38 9 58.3-61.3 59.8 7.89 7.58 0.111
7 0.37 2 61.3-62.5 61.9 3.24 19.12 0.500
 THE MONOD EQUATION I677

[CIA] CAON
N+I
where

CAO
-- N
\ I c C , max possible N' I
\
\
\
\
\
CC,optimum
------2. production r o t e of c e l l s

I
I
- 4-n

here
I washout maximum production
r a t e of cells

Fig. 4. Concentration vs. mean residence time behavior for substrate-limiting

Monod kinetics in a mixed-flow reactor where Cco = 0.

duction rate of cells, etc., are simple functions of k and of the ratio
CA 0 / CJ,.
To evaluate the kinetic constants from a series of steady-state
mixed-flow runs, rearrange eq. (10) to give
I/CA = (k/C.~,w)..r,n
- 1/CM ( 104

Plotting the concentration versus mean time data as in Figure 5,

fitting the best line through the data, and finding its slope and
intercept then gives the constants k and C , of the Monod equation.
Extension to operations where the feed contains cells, to optimum
operations of a number of interconnected flow reactors, and to
operations that use concentration and recycle of cells are all devel-
oped in Ref. 3.

111. PURE PRODUCT INHIBITION KINETICS

With sufficient food and a harmonious environment cells multiply

freely. However, no matter how much food is present there always
I678 LEVENSPIEL

Fig. 5 . Plot used to evaluate the kinetic constants for substrate-limiting Monod
kinetics in a mixed-flow reactor where Cco = 0. Data points are included for illus-
tration only.

comes a point where either the cells crowd each other out, or their
waste products inhibit growth. Hence Monod kinetics can be looked
upon as a special case of a more general rate form that includes
some sort of inhibition. This section develops the performance
expressions for the special case of sufficient food, or C, %- C M ,thus
for the rate expression of eq. (3). Here it is most useful to develop
all the equations in terms of C Rand Cc, not C,.

Batch or Plug-Flow Reactor

In terms of C Ror C c the performance expression is given as

f / , = T,, = 1y
cn dC,
('no
-
-
('CO

Inserting eq. (3) into eq. ( 1 I), writing all concentrations in terms of
Cc or CR,and taking the special case where n = 1 gives, on inte-
gration

When plotted, this equation gives an S-shaped curve whose shape

depends on Cco, as shown in Figure 6.
For n # 1, integration of eq. ( 1 I ) is messier and the final expres-
sion more complex; however, the shape of the concentration-time
curve will still be S-shaped, somewhat similar to Figure 6.
 THE MONOD EQUATION 1679

Mixed-Flow Reactor
The basic performance expression that relates input to output
from a mixed-flow reactor is still eq. (9). So inserting eq. ( 3 ) into it
and converting all concentrations into C Rwith eq. (4) gives
CR - CRO
kTm = (13)
(CR - CRO + [R/C]CCO)(l- cR/CR*Y
and for the special case where CcO= 0, or where the feed enters
free of cells
kTfn = (1 - CR/CR*)-' for kTff8> 1, Cco = 0 (14)
The properties of eq. (14) are displayed in Figure 7, which shows
that washout occurs at k ~ , =, ~1 , and that the maximum production
rate is dependent in a simple manner on CR* and n .
To find the kinetic constants from experiment first evaluate C R*
in a batch run using an excess of substrate A and letting r + 30.
Then rearrange eq. (14) to give
log Tj,, = - log k + n log [C.q*/(C,* - C R ) ] (15)
and plot as in Figure 8. The slope and intercept of the best line
through the data will then give the kinetic constants of the reaction.

medium Cco

Fig. 6. Concentration-time curve for pure product inhibition kinetics in a batch

or plug-flow reactor.
I680 LEVENSPIEL

CR
Points X and Y have the
same production rate

washout J ' n l o p t i m u m conditions

For n = I Topt= 2 Tw,,hout
'

Fig. 7. Concentration vs. mean residence time behavior for pure product inhi-
bition kinetics in a mixed-flow reactor where Cr,, = 0.

IV. SITUATIONS WHERE BOTH SUBSTRATE AVAILABILITY

AND PRODUCT INHIBITION AFFECT THE RATE

Here we must use the complete rate expression of eq. ( 2 ) , or

rC = [C/RlrR = kot,s*[CACr/(CA-k CM)]
where
kdl, = k(l - c,/c,x)n (16)
It turns out that it is impractical to use a batch or plug-flow reactor
to unravel the interacting effects of the four rate constants of this
equation, k , C,M,C R * ,and n, and these are best found in a mixed-
flow reactor. For this suppose we make a series of runs, each at
different C R , using a cell-free feed. From the reactor composition
we can find all the concentrations, rates, and T,,,values. Then the
performance expression of eq. (9) combined with eq. (16) and rear-
ranged gives
Trn = l / k o b s -k ( C A M / k o b s ) . ( 1 / C A ) with CCO= 0 (17)
Plotting the data as in Figure 9 then gives a family of curves, each
at different C R .This graph gives C,wand k(1 - CR/CR*)". Then,
knowing CR* (this is usually easy to evaluate beforehand) we can
 THE MONOD EQUATION 1681

log 7;
I
i.'
Fig. 8. Method for evaluating the rate constants for pure product inhibition
kinetics from experiments in a mixed-flow reactor where C,., = 0. Data points are
included for illustration only.

then find n and k from Figure 10; or else, if C,* is unknown, guess
it until you get a straight line on Figure 10.
The following example illustrates the procedure.

Example on Product Poisoning

Bazua and Wilke' reported the following data on the fermentation
of glucose ( A )to form ethanol ( R ) ,using Saccharomyces cerevisiae

slope =

intercept

CM

Fig. 9. Plot used to find the rate constants of the generalized Monod equation
from a set of runs in a mixed-flow reactor where C,.,, = 0. Data points are included
for illustration only.
I682 LEVENSPIEL

t find k f r o m t h i s intercept
7

Fig. 10. Evaluation of the toxic power n in the generalized Monod equation.
Data points are included for illustration only.

( 0 ,in a mixed-flow reactor, and in the presence of different levels

of ethanol. Find a rate equation to represent this fermentation in-
cluding the effect of inhibition by ethanol.

Solution
Using a portion of the reported data make the tabulation given in
Table 11. Plot these data as in Figure I 1 from which we find slopes
and intercepts and
C , , ,= 0.222 giliter
Also from the reported data we have
-
[RIG] = 3.90
Since an experimental value for CR* is not reported in the paper,
and is only said to be somewhere between 80 and 100, let u s find it
by the above recommended procedure. First guess CR*= 100. Then
tabulate as in Table 111. A plot as in Figure 12 does not give a
straight line. So try again with C,* = 90 and then 87.5. The latter
value gives a reasonably straight line. So from this figure we have
n = 0.41 (slope)
k = 0.42 (from intercept)
CR’.= 87.5 g/liter
 THE MONOD EQUATION I683

interceot I /

-4 5 0
I
I0
I
20
-
I
- (liter/grn)
=A

Fig. I I . Plot of T , vs.

~ I/CAfor the data of Table I1

The best-fit value reported by the authors is CR* = 93-94 @liter.

The final rate equation is thus, in terms of cell production,
YC = 0.42(1 - CR/87.5)0.41[CA/(CA + 0.222)]
o r in terms of alcohol production
rR = r c [ R / C ] = 1.64(1 - C,/87.5)0.41[CA/(CA -I-0.222)]
This rate equation is of the generalized Monod type, and it accounts
for both substrate availability and alcohol inhibition.

V. DISCUSSION

1) The similarity in shape of the graphs for substrate-limiting and

for product inhibition kinetics (compare Figs. I and 6 for plug flow,
Figs. 4 and 7 for mixed flow) may lead one to fit product inhibition
systems with the convenient Monod equation. The fit probably will
be good; however, extrapolation to new and different conditions
will lead to error because the logic in the use of this equation will
be quite wrong.
1684 LEVENSPIEL

TABLE I1
Bazua and Wilke's Mixed-Flow Reactor Data Reanalyzed

Reported From Figure I I

Calculated from
7 tic* reported data
(hr) (literlg) CIl [RlCI slope intercept

13.88 20.8
5.21
3.79 3.65 0.56 2.5
3.70 2.50
2.38 0.22

16.67 22.20
7.58
3.97 > 29.19 3.76 0.62 2.8
3.21
2.66 0.15

13.66 1 I .49]

26.04 12.05
14.37
13.44
8.13 4.88 1.60 7.2
10.42 2.08
9.92 2.08
7.25 0.17

One must first find which of these two factors is limiting and then
use the corresponding correct equation form.
2) Let us compare an expression proposed by Aiba et a1.8 for
substrate availability plus product inhibition kinetics with the gen-
eralized Monod equation proposed here.
Exponential-type equation
rr = kle-k2CR.[CACr/(CA+ C , v ) ] (18)
Generalized Monod equation
TC = k(1 - CR/CR*p[CAC,/(C, + C,v)] (19)
For given cell and substrate concentration, eq. (18) does predict a
slowing of the rate as the concentration of toxic product R rises.
But it also says that cell action will always continue no matter how
 THE MONOD EQUATION I685

intercept = log k = -0.377

k = 0.42
:_

slme = n = 0.41
--06

log
- -0 7
[k (I - 3
7
R

for cl,
--08

--09

-12 -10 -08 -06 -04 -02 0

l o g ( l - + )C
CR

Fig. 12. Plot of log [k(l - Cfi/Cfi*)n]V S . log ( I - CR/~R*).

high the concentration of R becomes. Thus in alcohol production

it says that even when the alcohol concentration is 20, 40, or 60%,
fermentation continues. This is not what is observed, hence eq. (18)
cannot be used for such situations.
As opposed to this, eq. (19) says that there is a definite upper
concentration limit for inhibitory product CR*above which fermen-
tation ceases.
The empirical constant n in eq. (19) accounts for the fact that one
can approach this limit in various ways: linearly ( n = I ) , rapid

TABLE 111
Evaluation of the Toxic Power, n
1 - 2 log 1 -_ 1 log[k(l -c,/c,*)n1=
CR CR* ( :I*) k ( l - Cfi/Cfi*)" log k +nlog(l - C R / C R * )
4.37 0.956 -0.0195 2.5 -0.3979
22.19 0.778 -0.1090 2.8 -0.4472
61.29 0.387 -0.4123 4.0 -0.6021
81.30 0.187 -0.7282 7.2 -0.8573
I686 LEVENSPIEL

initial drop in rate followed by a slow approach to CR*( n > I), or

vice versa ( n > 1). This is shown in Figure 13. If one does not
approach too close to C R * ,eqs. (18) and (19) with n B I could both
give approximately the same fit to data.
Holzberg et in effect, used the special case of eq. (19) where
n = I to fit their experimental findings.
3) In this paper we have used the normal language of reactor
design. The relationship to the corresponding language of biotech-
nology is as follows:
Dilution rate for mixed flow
D = l/Trp, = V/V
Specific growth rate
t-l = r c/ C c = k ( l - cR/cR*)nrcA/(cAC , , ) I +
Maximum specific growth rate in the presence of R

Pinax =--
rr
CC
k ( 1 - CR/C,*Y’ = k,,b\, CA * CJI
Maximum specific growth rate in the absence of R
~ l . 0= . k,
t . ( . / C (= C.4 * C,M, C, = 0
In simple situations either language can be used; however, in more
complex applications involving interconnected reactors, recycle, or
cell concentration, it probably is easier to follow what is happening

kobs

reaction stops

Fig. 13. Power n shows how the observed rate constant of the Monod equation
[eq. (2)] decreases as toxic product R rises.
 T H E MONOD EQUATION 1687

if one did not combine distinct quantities into an overall quality

such as p.
Extension to some of these operations can be found in Ref. 3.

Nomenclature
substrate, cells, rate-depressing product
concentration of A , C , and R in any convenient units
(such as moliliter: @liter)
concentration ofA, C , and R initially in a batch reactor
or in the feed entering a flow reactor
Monod or saturation constant: see eq. ( I ) (moliliter)
concentration of product R at which fermentation ends
dilution rate (hr-l)
flow or production rate (gihr or molIhr)
reaction rate constant: see eq. ( I ) (hr-l)
toxic power: see eq. ( I )
reaction rate (quantity formed/liter.hr)
time of reaction in a batch reactor (hr)
volumetric flow rate of fluid into and out of a steady
flow reactor (literihr)
reactor volume (liter)
yield, defined in eq. (4)

space time or residence time of fluid in a plug-flow

reactor (hr)
space time or mean residence time of fluid in a mixed-
flow reactor (hr)

The author would like to recognize Goran Jovanovic and Adonis Stephanakis for
deriving the expressions appearing in Figures 4 and 7.

References
I . J . Monod. Rechrrches sur la Croissnnce des Cultures Bacterienne (Hermann
et Cie, Paris, 1958).
2 . J. Monod, Ann. Re\.. Microbiol.. 3, 371 (1949).
3. 0. Levenspiel, Chemical Rerrctor. OfnnihooX (OSU Book Stores, Corvallis,
OR. 1979).
4. J. Monod. Ann. I n & / .Ptr.\tertr-. 7 9 , 39U (1950).
5 . A. Novick, and L. Szilard, Proc. Ntrrl. Actrd. Sci., 36, 708 (1950).
6. D. Herbert, S . I . C . Monograph. 12, 21 (1959).
7. C. D. Bazua and C. R. Wilke, Biotechnol. Bioeng. S y m p . , 7 , 105 (1977).
8. S. Aiba. M. Shoda, and M. Nagatani. Biorechnol. Bioeng., 10, 845 (1968).
9. I . Holzberg, R. Finn, and K. Steinkraus. Biotechnol. Bioeng., 9 , 413 (1967).

Accepted for Publication October 25, 1979