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Time-dependent kinetic models for

glutamic acid fermentation
Xue-Wu Zhang, * Tao Sun,* Zhong-Yi Sun, Xin Liu,* and De-Xiang Gu*
*Food Engineering Research Center, Zhongshan University, Guangzhou, Peoples Republic of
China: Educational College of Guizhou. Guiyang. Peoples Republic of China
A group of time-dependent kinetic models for glutumic acid fermentatiort are presented. These can divide cell
growth into three stages: the positive acceleration stage. negative acceleration stage, and equilibrium stage. The
substrate consumption is also divided into three stages: the negative acceleration state, positive acceleration
stage. and exhaustion stage. Product formation is also separated into three stages: the lag stage, positive
acceleration stage. and negative acceleration stage. By comparing data from the laboratory and facto9, good
agreement between experimental values and predicted values is found and in comparisons to reported models in
the documents about glutamic acid fermentation, the present models are most satisfactov. 0 I998 ElselGer
Science inc.
Keywords: Glutamic acid, fermentation kinetics. time-dependent models
The glutamic acid fermentation is a key process of sodium
glutamate production. The kinetic models play an important
role in monitoring and predicting fermentation processes. A
great number of dynamic models for the glutamic acid
fermentation have been reported.lm7 The common point for
the reported models is that these models basically existed in
the form of differential equations. These models are con-
stantly difficult to solve by integral form and only express
the implicit functions of time. They cannot directly reflect
the effect of fermentation time on cell growth, substrate
consumption, and product formation, the model-based real-
time control and monitor of the process variables (i.e., the
concentrations of biomass, substrate, and products) cannot
be determined. Especially for the glutamic acid fermenta-
tion, the results during the earlier period produce great
effects on results during the later period, so a predictive
model is greatly needed which can be used to estimate the
state of the later stage process based upon that of the earlier
stage process. Consequently, problems are found in a timely
manner. This guarantees the running of the process in the
desired route.
On the other hand, when simulating replacement of Ay/At
Address reprint requests to Dr. X.-W. Zhang, Zhongshan University, Food
Engineering Research Center, Guangzhou 5 10275, Peoples Republic of
Received I May 1997; revised 21 July 1997; accepted 5 August 1997
for dyldt, a greater discrepancy between the theoretical values
and the experimental values are produced; hence, an influence
on the accuracy of the model is observed. Moreover, the
structured model completely based upon mechanism is very
complicated and not as adaptable as expected, so to search for
or a new and simpler model structure which is suitable for
monitoring and predicting both laboratory-scale and factory-
scale fermentation processes, this becomes the urgent task of
researchers in biochemical engineering.
A fermentation process is a nonlinear and time-depen-
dent process; thus, the kinetic models describing the process
should also be nonlinear and time dependent. The objective
of this study is just an attempt to solve this problem. Based
upon the integral form of an existing model, a group of
empirically nonlinear and time-dependent kinetic models
for cell growth, substrate consumption, and product forma-
tion for the glutamic acid fermentation are proposed. By
these models, the detailed description of the glutamic acid
fermentation process is made. This lays the theoretical
foundation for the design, optimization, and scaleup of the
Theoretical aspects
The reported models for glutamic acid fermentation
(1) Cell growth models
dt =
Enzyme and Microbial Technology 22:205-209, 1998
0 1998 Elsevier Science Inc. All rights reserved.
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PII SOl41-0229(97)00168-3
dt I lrnK, + S
where Model (2) is just the well-known Monod equation.
(2) Substrate consumption models
dS 1 dX 1 dP
dt y,dt+y,dt
dS v,sx
dt K, + S
(3) Product formation model
dt= K,,, + S - %
Construction of time-dependent models Eq. (I) can
be integrated in the form:
= 1 - (X,/X,)[ 1 - exp (p&)
Starting with Eq. (6) and screening repeatedly, a group of
empirical, simple, and uniform equations are chosen to
describe the cell growth, substrate consumption, and prod-
uct formation for the glutamic acid fermentation:
X = XL exp ( - b,(t - tx) _ y) (7)
S = S, exp ( - b, (t - tJ) (8)
P = PL exp ( - bp(t - t,J - )
where X, S, and P are the cell, substrate, and product
concentrations respectively. X, is the limit concentration of
the cell (t = m); P, is the limit concentration of product (t =
~0); S, is the ideal concentration of substrate: t is the
fermentation time; t, is the time when X = 0; t, is the time
when S = St; t, is the time when P = 0. Usually, t,, ts, t,
5 0. If the inequality holds, it shows that the cell and
product concentration have an initial value (though very low
probably) and that the initial substrate concentration (t = 0)
tends to be low. The ideal substrate concentration (S,)
should be higher than this. All b and IE are constants which
are responsible for the convergent rate of curves, i.e., the
speed of change for X, S, and P.
By Eq. (7), the process of cell growth can be divided into
three stages:
PAS:0 5 t 5 tx +
NAS:t 2 tXrP
D t,,, then $5 e,
Here, PAS represents the positive acceleration stage in
which the cell grows with a positive acceleration; NAS
represents the negative acceleration stage in which the cell
grows with a negative acceleration; ES represents the
equilibrium stage in which the rate of the cell growth
(dX/dt) is less than or equal to a small and positive number
(e,). So to speak, the cell growth arrives at the equilibrium
under the ex level. The symbol D means is defined
By Eq. (8), the process of substrate consumption can be
divided into three stages (n, 2 1):
NAS:O 5 t I ts +
PAS:t 2 tsrp
NES:t I ts + Dt,,,, then S % es
If 0 < n, < 1, then there are two stages left: PAS (0 < t <
00) and NES.
Here, NAS and PAS mean the same as above. In NAS,
substrate decreases at a negative acceleration, and at a
positive acceleration in PAS. NES is the nearly exhausting
stage of substrate under the es level where es is a positive
Finally, by Eq. (9), product formation can be divided into
three stages:
PAS:0 5 t 5 tp +
NAS:t 2 tPrP
LS:O 5 t f t, + ( ln GL,l,,)..Dt,,,, then P 5
where PAS and NAS are the same as above. In PAS, the
product is formed at a positive acceleration, and at a
negative acceleration for NAS. LS stands for the lag stage in
which a little product is formed. The concentration of
product is not larger than a small and positive number (Ip).
In fact, the division of PAS and NAS can be understood
as follows. In the beginning of the fermentation, sufficient
substrate makes the cells grow at a positive acceleration and
the product accumulates at a positive acceleration after lag
time while the lower base number both for cell concentra-
tion and product concentration make the substrate decrease
only at a negative acceleration. As the fermentation goes on,
the increasing cell and product concentration enlarge the
needs for substrate so that substrate decreases at a positive
acceleration after a period of time. On the other hand, the
decreasing substrate limits the growth of cells and the
formation of product so that they have to slow down
gradually. The cell grows at a negative acceleration and
arrives at equilibrium in time. The product accumulates at a
negative acceleration during the middle or late stage of the
fermentation; thus, from the theoretical analysis, the present
models conform to the laws of the fermentation process and
are expected to describe the process of glutamic acid
fermentation well. Below are applications in production for
the proposed time-dependent models.
206 Enzyme Microb. Technol., 1998, vol. 22, February 15
Time-dependent kinetic models: X. W. Zhang et al.
Table 1 Parameter values for different models
Item Model
Laboratory data
Parameters Error
Factory data
Parameters Error
Models (X)
Models (S)
Models (P)
X,,, = 2.204
IL,,, = 0.4406
JL, = 0.20
KS = 0.70
X, = 3.7679
tx = -4
b, = 64.0077
n, = 1.4
Ye = 20.4166
Y, = 0.322
V,,, = 0.0123
K,,, = 0.3303
S, = 1.3236
t, = -4
b, = 7.45x 1O-5
n, = 2.8
V, = 0.0084
K,,, = 0.5676
a = 0.0129
PL = 1.0696
tp = -36
bp = 3191391.88
np = 3.0
X = 9.256
).I,,,, = 0.2072
)L., = 0.4146
KS = 1860.28
X, = 9.270
tx = -12
b, = 186557.009
n, = 4.5
T, = 0.4456
Y, = 0.0575
v, = 0.5545
K,,, = 7.245
S, = 125.984
t, = 0
b, = 2.29x 10m3
n, = 2
V,,, = 0.0286
K,,, = 2.278
a = 0.1380
PL = 10.7839
tp = -12
b, = 3393.02
np = 2.3
Units: X, S, P, X,, S,, s, X,,,, KS, K,,, are all in g I-; r.+,, (h-l), V,,,(h-),Y, (g g-,Y, (g 9-l)
Results and discussion
Two sets of data are used for simulation. One comes from
a laboratory 2 and the other from a factory.8 Cell density
was measured turbidometricahy at 610 nm and converted
into a cell mass concentration using a calibration curve. The
concentration of glucose was measured by the glucose
oxidase method (Wako Pure Chemical Industries, Japan)
and the concentration of glutamic acid by the calorimetric
method using glutamate dehydrogenase (Boehringer Mann-
heim, Mannheim, Germany). The total sugar concentration
was measured by the phenol sulfuric acid method.
The generalized least square method is applied for the
parameter estimation in the different models. The simula-
tions are evaluated by the fitting error:
\ n I
where Yi and Yi* are the original and the predicted value,
respectively, and n is the number of data used.
The values of parameters and the fitting errors for
different models are listed in Table 1. The simulation curves
with different models for cell, substrate, and product are
illustrated in Figures IA-IF.
It can be seen from Table I that the fitting errors for
different models that fit the data from laboratory and factory
can be sequenced as below:
For cell growth: Model (2) > Model (1) > Model (6)
For substrate consumption: Model (3) > Model (4)
> Model (8)
For product formation: Model (5) > Model (9)
Thus, the time-dependent Models (7)-(9) presented in this
paper have the smallest error in all the situations. Obvi-
ously, the simulation curves with different models (Figures
lA-1F) show that the time-dependent models are best.
Meanwhile, the critical values appearing in the inequal-
ities (lo)-( 15) can be calculated. For the laboratory: tXtp =
9.3134, t+ = 48.7557 (e, = 1% X, = O.O25g/L), tstp =
21.4537, t,, = 3 1.3473 (e, = 20% S, = 0.26g/L), tptp =
30.4596, t,p = 7.4606 (lr = 3% P, = O.Olg/L) For the
factory: tXtp = 2.1881,t,, = 15.7374(ex = 1% X, =
0.09gU ts,p =
24.48/L), t,, =
14.7786,&? = 26.7780 (es = 20% S, =
17.3002. t,p = 6.0822 (It, = 3% P, =
O.l4g/L) Here, S,, X,,,, and P, are the initial concentration
of substrate (t = O), and the maximum concentration of cell
and product, respectively.
A detailed description of the glutamic acid fermentation
in the laboratory or factory (corresponding to the figures in
parentheses) can be made:
1. The cell grows at a positive acceleration when 9.3 134
(2.1881) Th ago, and- this time backward grows at a
negative acceleration. The cell growth will arrive at an
equilibrium under the level of ex = 1% X, = 0.025g I-
(e, = 1% X, = 0.09 g l-l), i.e., after 48.7557 (15.7374)
h, dX/dt < 0.025 g I- hh (0.09 g I- hh). In fact
within the interval of 28 (15) - 32 (16) h, the increasing
rate of cell growth AX/At is 0.033 (0.1) g 1-l hh =
1.3% X, (1 .l% X,), really near to equilibrium.
The substrate decreases at a negative acceleration when
21.4537 (14.7786) h ago, and this time backward de-
creases at a positive acceleration. After 31.3473
(26.7780) h the concentration of substrate will be lower
Enzyme Microb. Technol., 1998, vol. 22, February 15
0 4 8 I.2 16 20 24
0 4 8 I2 16 20 ti
Time t(h)
Time tfh)
Fi gur e 1 Simulation curves of the glutamic acid fermentation by using different models to fit the experimental data. For A-C, the data
are from a laboratory. For D-F, the data from a factory.*
than the level of es = 20% S, = 0.26 (24.4)g 1-l. In fact, fermentation is about 7.4606 h (6.0822 h). Within this
the concentration of substrate at t = 32 h (25h) is equal
to 0.25 (28) g 1-l =
period of time, the concentration of product is under the
18.94% S, (22.95% S,), which is level of 1, = 3% P, = 0.01 (0.14) g 1-l. In fact, the
basically near the predicted level. concentration of product at t = 8h (5 h) is equal to 0.017
3. The lag time for product formation in the glutamic acid (0.11) g 1-l = 5.6% P, (2.4% P,) which is basically
208 Enzyme Microb. Technol., 1998, vol. 22, February 15
near the predicted level. As the fermentation goes on, the
product accumulates at a positive acceleration 30.4596
(17.3002) ago, and this moment later accumulates at a
negative acceleration.
From the above research, it can be seen that the structure
of time-dependent models is adaptable both to the labora-
tory and factory. Different processes simply correspond to
different values for parameters. In the course of the glutamic
acid fermentation, the results in the beginning of the
fermentation have a great impact on those at the end of the
fermentation; hence, the time-dependent model can be used
to predict what happens toward the end of the fermentation
based upon results from the early fermentation so that
problems can be discovered in a timely manner and some
measures can be taken to assure that the fermentation
proceeds as expected.
Apparently looking at the expression of time-dependent
models (7)-(9), the interrelations among cell, substrate, and
product cannot be displayed in the models; however, after
mathematical calculations, the time-dependent models can
be transferred into the form of a differential equation which
implies interactions among cell, substrate, and product. The
transferred equations can be written as
(Jb/2)K~ (.fPf2)P.s
pp = .fx + OP - t?i)lJ cY +fs + CtfJ - t.dl%
(fs/2)px (fs/2)l-b
ps = .fY + (ts - tx)l-h +fP + (ts - t,)l-b
1 dX
where kx = x x is the specific growth rate of cell, kp =
IdP 1 dS
~dt is the specific formation rate of product, ks = -- -
S dt
is the specific consumption rate of substrate, and fx = n,
ln(X,/X), fp = np ln(P,/P), fs = n,ln(S,/S).
Thus, the time-dependent models (7)-(9) presented in
this paper can describe the interactions among X, S, P, k,_
ps, and kp by their derivative equations (16)-(18).
Beginning with the integral form of known equation, a
series of time-dependent kinetic models for the glutamic
acid fermentation have been proposed in this study. Com-
pared to the reported models, the present models have the
following advantages: (1) They are the explicit functions of
fermentation time; hence, they can directly reflect the effect
of time on cell growth, substrate consumption, and product
formation. In other words, realtime monitoring and control
can be realized; (2) They can make a more detailed
description of the fermentation process, i.e., the positive or
negative acceleration stage and the equilibrium time of the
cell, nearly exhausting time of substrate and the lag time of
product under the given level of critical value can be
determined, which is very important in real production; (3)
Time-dependent kinetic models: X. W, Zhang et al.
They have a simple and unified structure of model: Y =
aexp [ -b(t - c)], where n < 0 for cell growth and product
formation and n > 0 for substrate. It is easy to estimate the
parameters. In this study, the fitting results are best; (4)
They can also be transferred into the form of a differential
equation as in the reported models. The interactions among
cell, substrate, product, and their specific rate can be
described while this makes one of the advantages of the
known differential models disappear; however. the pre-
sented models need further verification and development.
cl = Constant in Eq. (51 (g I-)
;&.fP 1
i, P,. P, =
s, S,, S, =
fXCP> CstP~ fP,P =
x, x,, x, =
yc7 YM =
fJ,xs I.9 CLP =
Fitting error
Functions in Eqs. ( 16)-( 18)
Constant in inequality ( 15) (g I- )
Concentration. limit concentration, and
maximum concentration of product (g l--l)
Concentration, ideal concentration, and
initial concentration of substrate (g I- )
Fermentation time (h)
Constants in inequalities (IO), (l2), (14)
Concentration. limit concentration. and
maximum concentration of cell (g I-)
Constants in Eq (3) (g g- 1
Specific growth rate of cell (h- ), specific
consumption rate of substrate (h-l), spe-
cific formation rate of product (h-l)
Constants in Eqs. (7)-(9)
Constants in inequalities (1 I), (13) (g I--)
Constants in Eqs. (21, (5) (g I-~)
Constants in Eqs. (7)-(9)
Constants in Eq. (7)~(9) (h)
Constant in Eq. (4) (h-l)
Maximum growth rate (h- )
b,, 6,. b, =
z;, ;m =
ilx, n,, np =
rv !u f,, =
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