Keehyun Han* and Octave Levenspiel
Department of Chemical Engineering, Oregon State University, Corvallis, Oregon 9733 1
Accepted for publication August 6, 1987
A generalized form of Monod kinetics is proposed to ac count for all kinds of product, cell, and substrate inhibi tion. This model assumes that there exists a critical inhibitor concentration above which cells cannot grow, and that the constants of the Monod equation are func tions of this limiting inhibitor concentration. Methods for evaluating the constants of this rate form are presented. Finally the proposed kinetic form is compared with the available data in the literature, which unfortunately is very sparse. In all cases, this equation form fitted the data very well.
INTRODUCTION
Although microbial growth is a very complex phenome non, the overall growth can often be regarded as a single chemical reaction with a simple rate expression. Many equations are used for this purpose. Among these, the sim plest and most popular is the one proposed by Monod who assumed that a single essential substrate is the growth limiting factor. I Monod kinetics can be expressed as
Substrate (A) 
Cells (C)
more Cells
(C) + Product (R)
r,
=
k
cell mass produced
where r,
is the growth rate of cells; k is the reaction rate
constant; CA is the substrate concentration; C, is the
cell
concentration; and C, is the Monod constant. Unfortunately, there are often inhibitory effects in the real world. In this article, we propose a generalization of the Monod expression which takes into account inhibition effects caused by high concentration of either substrate, or cell, or product, or other inhibitory substance. This pro posed expression is
where C, is the inhibitor concentration, C: is the critical inhibitor concentration above which reaction stops, and n and m are constants.
* Present address: Biochemical Engineering Program, School of Engi neering, University of California, Irvine, CA 92717.
For substrate inhibition:
for product inhibition:
for cell inhibition:
In the extreme where C, G C ,: the above eqs. (2)(5) all reduce to the simple Monod expression of eq. _{(}_{1}_{)}_{.}
HOW TO EVALUATE THE CONSTANTS OF THE GENERALIZED MONOD EQUATION FOR PRODUCT OR CELL INHIBITION
As with Monod kinetics, the constants in eq. (2) with
C, =
C ,
or C, =
C ,
can be evaluated from a Lineweaver
Burk plot of C,/r, vs l/CA.Thus inverting eq. (2) gives

r,
C,(1
 c,/c:)m1
k(l  C,/Cf)"
CA
1
k(l  C,/CF)"
This expression can account for and represent the six com mon patterns of inhibition illustrated by the Lineweaver Burk plot of Figure _{1}_{:}
(a) represents noncompetitive inhibition, where n > 0 and m = 0;
(b) represents competitive inhibition, where n = 0 and m < 0;
(c) represents generalized uncompetitive inhibition, where n>m>O;
(d) represents generalized uncompetitive inhibition, where m>n>0;
Biotechnology and Bioengineering, Vol. 32, Pp. 430437 (1988)
0
1988 John Wiley & Sons, Inc.
CCC 00063592/88/04043008$04.00
Calculate C,,, from eq. (17), and plot as in Figure 6. The slope and intercept on this graph gives C, and m.
Substrate Inhibition: Example 1
Uemura and coworkers6 studied the microbial attack on
n pentane by a pentaneconsuming bacterium (JTB27 1). The results were read from Figure 5 of Edwards' paper,7 and are shown in Table 11. A plot of these data points in Figure 7 clearly shows substrate inhibition since the rate of reaction slows at high substrate concentration. When C,
2
65 mmHg, assume that C, S CMobs, in which case eq. (3)
is well approximated by eq. (15). It will be shown later that this is a reasonable assumption. Following the procedure outlined above, we first make the plot of Figure 8, from which we find C,* = 700 mmHg, k = 0.117 h', and n = 1.82. CMobsvalues are then calcu lated from eq. (17) and listed in Table 111. Finally, the plot of Figure 9 gives C, = 30 mmHg and m = 66.8. At C, =
65 mmHg, we find CMobs= 0.045 mmHg, hence the as
sumption that C, S CMobsand m > 0 is well satisfied. The final result is then
0: data from Uemura et aI6 

eq. 18, best fit 
of 
the 
general Monod equation
_{0}_{'}
0
1
1
200
1
400
1
600
_{p}_{a}_{r}_{t}_{i}_{a}_{l} pressure of npentane (mm Hg)
Figure 7. Data of ref. 6 clearly shows that high concentration of npentane, the substrate, inhibits the reaction; see Table _{1}_{1}_{.}
(18)
This expression fits the reported data well, as shown in Figure 7.
2.8
3.0 

0.4 
0.2 
0 
Figure 8. This graph gives k, n, and C,* for tion; data of ref. _{6}_{.} 
npentane substrate inhibi 

( ::)I 
0 

In 

Table III. 
C,, 
value extracted from ref. 6. 

Figure 6. 
For substrate inhibition m and C, are evaluated at low C, 

_{T}_{a}_{b}_{l}_{e} _{1}_{1}_{.} Experimental result of ref. 6 taken from Figure 5 of ref. 7. 5.000 


6.875 


10.938 


15.313 


19.688 
0.091 1 
4.32 

5.000 6.875 
0.0227 0.0378 
25.625 


10.938 
0.0564 

15.313 
0.0792 

19.688 
0.0911 
Substrate Inhibition: Example 2 

25.625 
0.0992 

30.625 
0.0992 
Kortan* reported substrate inhibition by n butanol to 

65.000 1oo.OOo 135.625 
0.0991 0.0817 0.0786 
Arthrobacter AK 19. The experimental results of Table IV are from Figure 2 from the article by Wayman and Tseng.' As in the previous example C,*, k, and n values are ob 

199.375 
0.0633 

268.7 50 
0.0489 
tained from Figure 10. In this example, we assume that eq. (3) is well approximated by eq. (15) when C, 3 
HAN AND LEVENSPIEL: EXTENDED MONOD KINETICS
433
product inhibition, as shown in Table VI. However the ki netic form proposed here should be useful for all forms of inhibition. In addition if there are no inhibitory effects (n _{=} 0 and m = 0), the generalized equation reduces to simple Monod kinetics. Finally, the key characteristic of the proposed model is its assumption that there exits a critical inhibitor concentra tion above which reaction ceases. This is what actually happens in many, if not most situations. Many of the prod uct inhibition models incorporate a critical inhibitor con centration. On the contrary, for substrate inhibition only Wayman and Tseng’s model includes this feature. Conse quently, those models which do not have a critical inhibitor
concentration predict that inhibitor concentration should be infinite to stop the reaction.
Substrate Inhibition
Edwards’ compared the goodness of fit of five kinetic models for substrate inhibition to the reported data of Uemura et aL6 Table VII lists these models, Table VIII gives the bestfit value of the constants of these models, and Figure 13 displays the results as dotted or dashed lines. Also included in this figure is the curve from the model developed here, or eq. (18). This figure shows that one should be able to more clearly discriminate between models at high substrate concentration.
_{T}_{a}_{b}_{l}_{e} _{V}_{I}_{.}
Kinetic models for inhibition; equations adapted from p. 256 and p. 259 of ref. 10.
Equation 
Reference 

For substrate inhibition: 

rC k  
1 

Andrews and Noack 

CC 
+ cM/cA 
+ CAIK/ 

Webb 

Yano and coworkers 

Aiba and coworkers 

‘c = k [exp(CA/Kl)  exp(CA/C~)] 
Teissiertype 

CC 

(u is ionic strength) 
Webb 

Wayman and Tseng 

‘c =k 
CA 
 K1(CA CJ, 
when CA> Ci 

CC cM 
+ CA 

For product inhibition: 

’c=k 
CA 
(1  KCR) 
Dagley and Hinshelwood 

CC CM + CA 

rC  k  CC  
Holzberg and coworkers 

K(CR  KI) 

%=k CC ( 
I 
 
Ghose and Tyagi 

‘c=k 
CA 
exp( KCR ) 
Aiba and coworkers 

CC CM + cA 

rc =k 
CA 
KI 
Jerusalimsky and Neronova 

_{C}_{c} CM 
+ CA Kl + CR 

‘c = k(1 3T5L 
Bazua and Wilke 

cc 
CM + CA 

Levenspiel 
HAN AND LEVENSPIEL: EXTENDED MONOD KINETICS
435
0.10
0.08
0.06
0.04
present
model predicts
this limiting point
0.02
0
0
200
400
600
700
Figure 13.
partial pressure of
npentane (mm Hg)
Fit of various proposed substrate inhibition equations of
Table VII to the data of ref. 6. Note that data at high pentane concentra
tions would sharply discriminate between models.
NOMENCLATURE
CA 
substrate concentration (substrate/volume) 

CC 
cell concentration (cell/volume) 

c, chf 
inhibitor concentration (inhibitor/volume) Monod constant 

Chf, 
observed value of C, 

Clf 
product concentration (product/volume) 

_{C}_{*} 
critical concentration above which reaction cannot pro 

iandj 
ceed; C: for inhibitor, C: for cell, Ci for substrate, C,* for product positive integers 

k 
reaction rate constant (time') 

k*s 
observed value of k 

K, K,, 
and Ks 
constants in Table VI 
and Table VII 
nandm 
constants 

rC 
rate of cell formation (cell mass/volume . time) 
This work was carried out under NSF Grant no.: CBT851.8737.
References
1. J. Monod, Ann. Rev. Microbiol., 3, 371 (1949).
2. C. N. Hinshelwood, The Chemical Kinetics of the Bacterial Cell (Clarendon, Oxford, 1952).
3. C. D. Bazua and C. R. Wilke, Biotechnol. Bioeng. Symp., 7, 105 (1977).
4. 0. Levenspiel, Biotechnol. Bioeng., 22, 1671 (1980).
5. S. Aiba, M. Shoda, and M. Nagatani, Biotechnol. Bioeng., 10, 845 (1968).
6. N. Uemura, J. Takahashi, and K. Ueda, J. Ferment. Technol., 47, 220 (1969).
7. V. H. Edwards, Biotechnol. Bioeng., 12, 679 (1970).
8. K. Kortan, M. S. thesis, University of Toronto, Ontario, Canada,
1972.
9. M. Wayman and M. C. Tseng, Biotechnol. Bioeng., 18, 383 (1976).
10. A. Moser, in Biotechnology, Volume 2, H. Brauer, Ed. (Verlag Chemie, Weinheim, 1985).
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437