Chemical Engineering Science 56 (2001) 5103–5110

www.elsevier.com/locate/ces

Parametric sensitivity in bioreactor: an analysis with reference to

phenol degradation system
S. Duttaa , R. Chowdhuryb , P. Bhattacharyaa; ∗
a Chemical Engineering Department, Jadavpur University, Calcutta-700032, India
b Food Technology and Biochemical Engineering Department, Jadavpur University, Calcutta-700032, India

Received 16 September 2000; received in revised form 6 February 2001; accepted 29 March 2001

Abstract
Sensitivity analysis of a biochemical system has been made in order to determine a priori a parametrically sensitive regime. A
pH-uncontrolled aerobic phenol degrading reactor using a mixed population has been chosen as a representative system. Normalized
objective sensitivities of the minimum pH have been determined with respect to several input parameters. Critical points at which
the system becomes sensitive simultaneously with respect to all input parameters have been determined. ? 2001 Elsevier Science
Ltd. All rights reserved.

Keywords: Bioreactors; Stability; Parametric sensitivity; Sensitivity analysis; Mathematical modelling; Simulation

1. Introduction that more such studies should be extended to biore-

Studies on parametric sensitivity of complex reaction
systems involving heat e6ects (Morbidelli & Varma,
1986a,b, 1989; Chemburkar, Morbidelli, & Varma, 1986;
Tambe, Inamdar, Bandopadhyay, & Kulkarni, 1991; Va-
jda & Rabitz, 1992; Heiszwolf & Fortuin, 1996; Juncu,
Brouwer, & Mulder, 1996; Baptista & Castro, 1996;
Varma, Morbidelli, & Wu, 1999; Paladino & Ratto,
2000) have opened up a new avenue towards getting
more insight to reactor system. From process engineer-
ing point of view the main objective of such studies
has been oriented towards safety aspects of reactors by
ascertaining a priori the regions of parametric sensitiv-
ity. Parametric sensitivity in bioreactor is an interesting
phenomenon on which contributions in the literature
are rather limited. The worth mentioning work in bio-
chemical engineering area is that of Rodrigues and Filho
(1996), who have done the sensitivity analysis of main
process variables for the production of penicillin. It is felt
∗ Corresponding author. Tel.: +91-33-4832378; fax: +91-33-
4734266.
E-mail address: pinaki che@yahoo.com (P. Bhattacharya).
actor system where reaction engineering behaviour is
highly inDuenced by microenvironment. In this case,
there is every possibility that one of the elements con-
stituting the microenvironment may be parametrically
sensitive.
The present investigation deals with phenol degrada-
tion system as a model biochemical system and Mor-
bidelli and Varma generalized criterion (1986a, 1986b,
1989) has been applied to perform parametric sensi-
tivity analysis using the pH-minimum as the objective
function. A deterministic mathematical model has been
developed to predict the change of pH, concentrations
of biomass and substrate with respect to the propagating
reaction time. The e6ect of pH and the inhibitory e6ect
of substrate, namely phenol, on speciEc growth rate has
also been incorporated by coupling polynomial function-
ality of the former (Lallai, Mura, Miliddi, & Mastinu,
1988) with the Haldane-type kinetic expression (Lallai
& Mura, 1989b). It is expected that such performance
of sensitivity analysis to know the parametric sensitive
region in bioprocess engineering prior to real process
run will be important from reactor operation point of
view.

0009-2509/01/$ - see front matter ? 2001 Elsevier Science Ltd. All rights reserved.
PII: S 0 0 0 9 - 2 5 0 9 ( 0 1 ) 0 0 1 8 7 - 7
5104 S. Dutta et al. / Chemical Engineering Science 56 (2001) 5103–5110

Table 1 and the substrate balance equation is

Values of di6erent parameters
dC dX=dt
Parameters Values =− : (5)
(Numerical=in function form) dt yx=s
c1 −0:045C0 + 4:199
c2 0.566C0 − 51:619
The values of m and yx=s are given in Table 1.
c3 −1:781C0 + 160:010 An empirical equation showing the rate of change of
e 0:002C0 − 0:073 pH with respect to both biomass and substrate concentra-
f −0:002C0 − 0:193 tion has been proposed as follows:
Ki 1503.400
KS 57.350 dpH
m 207.170 − = X e C f (6)
p −0:00036 dt
q 0.1548
r −4:2552 The values of ; e and f have been evaluated through
yx=s −0:234(X0 =C0 ) + 0:0011 the nonlinear regression analysis (regression coeH-
 1.000 cient: 0.9999) of the experimental data (Lallai & Mura,
max 0.088 1989a,b). Table 1 shows the values of ; e and f.
By introducing dimensionless parameters X ∗∗ ; X0∗∗ ;
C , C0∗∗ ; ; ; ;  and lag Eqs. (3) – (6) reduce to the fol-
∗∗

lowing dimensionless form:

2. Theoretical analysis For 0 6  6 lag ,

d
Coupling the polynomial functionality of system pH = ; (7)
d
with Haldane-type kinetics, the speciEc growth rate of
biomass () can be represented as follows: where  = a1 =C0∗∗ and a1 = mX0 tmax =C0; max .


max C0 Again, for  ¿ lag ,
= (c1 pH2 + c2 pH + c3 ):
KS + C0 + (C02 =Ki )
d
(1) = (1 + ){c1 (pH0 )2 + c2 (pH0 ) + c3 }
d
The values of max ; KS and Ki have been taken from = (1 + )(c4 2 + c5  + c3 ); (8)
Lallai and Mura (1989b) and the coeHcients c1 ; c2 and
c3 have been determined by nonlinear regression analysis


(regression coeHcient: 0.9982) of the experimental data d d
(Lallai & Mura, 1989a,b). = ; (9)
d d
tlag may be considered as a polynomial function of
substrate concentration and the relationship is given by
d
= − (1 + )e (1 − )f ; (10)
tlag = pC02 + qC0 + r: (2) d

The coeHcients p; q and r have been determined by non- where

linear regression analysis (regression coeHcient: 0.9997).
Table 1 shows values of all these parameters and coeH- c4 = c1 (pH0 )2 ; (11)
cients of Eqs. (1) and (2).
Now, the initial conditions of the bioreaction system
may be viewed as follows: c5 = c2 pH0 ; (12)
At t = 0,

X = X0 ; C = C0 and pH = pH0 : max C0 tmax

= ; (13)
For 0 6 t 6 tlag , the substrate balance equation is C0 + KS + (C02 =Ki∗∗ C0; max )

dC Ki
− = mX0 : (3) Ki∗∗ = ; (14)
dt C0; max
For t ¿ tlag , the biomass balance equation is
dX a2 X0∗∗
= X (4) = ; (15)
dt C0∗∗
 S. Dutta et al. / Chemical Engineering Science 56 (2001) 5103–5110 5105

 Table 2
X0; max
a2 = ; (16) Expressions used for evaluating &i for various parameters %i as
C0; max yx=s deEned in Eq. (28)

 = a3 (X0∗∗ )e (C0∗∗ )f (17) %i &i

and (e − 1)(1 + )(e−1) (1 − )f

X0∗∗ −
X0∗∗ (c4 2 + c5  + c3 )
(X0; max )e (C0; max )f tmax
a3 = : (18) C0∗∗
(1 + )(e−1) (1 − )f '
pH0 −
C0∗∗ (c4 2 + c5  + c3 )
The di6erential equations (8) – (10) have been solved nu- (1 + )(e−1) (1 − )f (
Ki∗∗
merically using the fourth order Runge–Kutta method Ki∗∗ (c4 2 + c5  + c3 )
with the following reoriented initial conditions:
At  = 0,
 = 0; =0 and  = 1:0: The initial conditions are
At  = 0 ,
@
$ = 1; s%i 0 = = 0;
3. Sensitivity analysis @%i
where, %i is one element of parameter vector
.
Sensitivity analysis of the system during the exponen-
tial phase of microbial growth has been done following
the method of Morbidelli and Varma (1986a,b, 1989), 4. Determination of sensitivities
Chemburkar et al. (1986) and Tambe et al. (1991). The
governing pH and substrate concentration equation in the Normalized objective sensitivity has been determined
general form may be written as using the following steps:
d {(1 + )e−1 (1 − )f }
= Q(
; ; ; ) = − ; (19) 1. Eqs. (20), (21) and (24) have been solved using the
d {(c4 2 + c5  + c3 )}
fourth order Runge–Kutta method until the value of
where
is the vector of model physicochemical and input ’’ attains its minimum. The corresponding values of
parameters. C0∗∗ ; X0∗∗ ; Ki∗∗ have been considered to be the ∗ and $∗ have been determined.
elements of parameter vector
: 2. sJ%0 given by Eq. (25) has been determined using the
DeEning, = =, Eq. (19) becomes value of $∗ with the help of the following equation:
d (1 + )e−1 (1 − )f sJ%∗ 1
=− ; (20) sJ%0 = = : (26)
d (c4 2 + c5  + c3 ) $ ∗ $ ∗
d 1
= : (21)
d  3. Objective sensitivity for each %i has been determined
using the following equation:
Now, letting  ∗
@Q (1 + )e−1 (1 − )f (c6  + c5 ) s%∗ i = s%i 0 sJ%0 + &i sJ% d; (27)
H =− =− ; (22) 0
@ (c4 2 + c5  + c3 )2
where
where
@Q
&i = for  ∈ (0 ; 1): (28)
c6 = 2c4 ; (23) @%i

the adjoint equation takes the following form: The expressions for ‘&i ’ corresponding to each %i have
been shown in Table 2.
d$ Substituting the expression for sJ% from
= H$ for  ∈ (0 ; ∗ ); (24)
d Eq. (25), Eq. (27) reduces to
where  ∗ 


sJ% s%∗ i = s%i 0 sJ%0 + &i $ d sJ%0 : (29)

$ = (25) 0
sJ%0
and 0 is the value of  at the onset of exponential phase 4. Normalized objective sensitivity S%∗i for each input
and ∗ refers to the value of the same at  = min . parameter, %i , has been evaluated using the following
5106 S. Dutta et al. / Chemical Engineering Science 56 (2001) 5103–5110

relationship: Substituting the value of s%∗ i from Eq. (31) into Eq. (30)


∗ one gets
d(ln ∗ ) %i d
S%∗i = = ∗ : (30)

d(ln %i )  d%i %i
∗
S%i = ∗ s%∗ i : (32)

It is to be noted that the objective sensitivity, s%∗ i , of any
model response with respect to any of the input parame-
ters, %i , is nothing but the derivative of the response with
respect to that particular %i . 5. Results and discussions
Therefore, in this case
In Figs. 1 and 2 the simulated concentration proEles
d∗ of biomass and substrate against time have been plotted.
s%∗ i = : (31)
d%i In Fig. 3 values of pH of the fermentation broth have
been plotted against reaction propagation time. The cor-
responding experimental data (Lallai & Mura, 1989a)
have been superimposed on the same plot. From the
close observation of the Egures, it is evident that the
simulated predictions of the present model incorporat-
ing Haldane’s substrate inhibition kinetics along with a
polynomial functionality of pH are in good agreement
with the experimental observations (index of correla-
tion for the proEles of biomass concentration, substrate
concentration and pH are 0:9583; 0:9066; 0:9053, respec-
tively).
pH sensitivity
In Figs. 4 –9, minimum-pH normalized objective sensi-
tivities, S%∗i with respect to the parameters, C0∗∗ ; X0∗∗ ; Ki∗∗
have been plotted as a function of , the dimensionless
group analogous to the Semenov number.
It appears that in each curve the critical (i.e., c ),
the location of maximum sensitivity, is the same for any
choice of parameter %i . This observation ensures the gen-
eralized nature of the adopted criterion because the fer-
menter pH becomes sensitive to all the model parameters
simultaneously. Thus, the parameter range characterized
Fig. 1. Simulated (—) and experimental (4) proEle of by ¿ c may be called ‘generalized region of para-
biomass concentration against time during exponential phase for
C0 = 280 mg dm−3 .
metric sensitivity’.
Some deEnite trends may further be noted from
Figs. 4 –9. These are as follows:

(1) For the present study, c values remain independent

of the choice of parameters.
(2) For Exed values of X0∗∗ (0:744) and Ki∗∗ (5:370), val-
ues of c decrease from 3.564 to 4:145 × 10−3 with
the increase of C0∗∗ from 0.214 to 1.000 (Figs. 4 and
5) and values of S%∗i also decrease with the increase
of C0∗∗ . This may be due to the fact that at higher
concentration of substrate the system becomes less
sensitive towards any of the components of input
vector,
.
(3) For Exed values of X0∗∗ (0.744) and C0∗∗ (0.536),
values of c remain constant with the variation of
Ki∗∗ in the range of 1:0 × 10−5 –100 (Figs. 6 and
7). This may be due to the fact that whether the
Fig. 2. Simulated (—) and experimental (4) proEle of sub- bioreaction is inhibited or uninhibited by the sub-
strate concentration against time during exponential phase for strate, the region of parametric sensitivity remains
C0 = 280 mg dm−3 . una6ected.
 S. Dutta et al. / Chemical Engineering Science 56 (2001) 5103–5110 5107

Fig. 3. Simulated (—) and experimental (4) proEle of pH against time during exponential phase for C0 = 280 mg dm−3 .

Fig. 4. Normalized objective sensitivity S%∗i as a function of for input parameter vector
with X0∗∗ ; C0∗∗ ; Ki∗∗ as parameters.

(4) The value of c decreases (3:205 × 10−2 –1:426 ×
10−2 ) with the increase of X0∗∗ in the range of 0.388–
1.000 (Figs. 8 and 9) if the values of C0∗∗ (0.536) and
Ki∗∗ (5.370) is kept constant. This may be due to the
fact that with the increase of inoculum concentration,
the biosystem becomes less sensitive to the biomass
concentration.
(5) In all Egures excepting that for Ki∗∗ = 1:0×10−5 (Fig.
6), the sensitivity curves corresponding to parameter
X0∗∗ lie at the highest level and those for Ki∗∗ lie at the
lowest one. The curves corresponding to C0∗∗ remain
at the medium level. However, in Fig. 6, X0∗∗ curve
lies at the lowest level, Ki∗∗ curve at the middle and
C0∗∗ curve reserves the highest position. This implies
that all systems (excepting that for Ki∗∗ = 1:0 × 10−5 )
show sensitivity towards X0∗∗ ; C0∗∗ and Ki∗∗ in the
descending order of magnitude. However, Fig. 6 is
the only exception. In this case, the value of Ki∗∗ is
1:0 × 10−5 . Therefore, the system is highly inhibited
by substrate. Hence, the sensitivity of the system
towards C0∗∗ and Ki∗∗ increases and exceeds the value
corresponding to X0∗∗ .
5108 S. Dutta et al. / Chemical Engineering Science 56 (2001) 5103–5110

Fig. 5. Normalized objective sensitivity S%∗i as a function of for input parameter vector
with X0∗∗ ; C0∗∗ ; Ki∗∗ as parameters.

Fig. 6. Normalized objective sensitivity S%∗i as a function of for input parameter vector
with X0∗∗ ; C0∗∗ ; Ki∗∗ as parameters.

Notation q coeHcients used in Eq. (2), (mg dm−3 )−1 (ks)

r coeHcients used in Eq. (2), ks
C substrate concentration, mg dm−3 s%∗ i Erst order objective sensitivity of min to pa-
C ∗∗ = C=C0; max rameter %i
C0∗∗ = C0 =C0; max S%∗i normalized Erst order objective sensitivity of
c1 ; c 2 ; c 3 dimensionless parameters used in Eq. (1) min to parameter %i
e; f dimensionless parameters used in Eq. (6) t time, ks
Ki inhibition constant, mg dm−3 tlag lag time, ks
KS saturation constant, mg dm−3 X biomass concentration, expressed as O.D.
m speciEc maintenance rate, mg dm−3 (ks)−1 X ∗∗ = X=X0; max
p coeHcients used in Eq. (2) (mg dm−3 )−2 X0∗∗ = X0 =X0; max
(ks) yx=s = yield factor (mg dm−3 )−1
 S. Dutta et al. / Chemical Engineering Science 56 (2001) 5103–5110 5109

Fig. 7. Normalized objective sensitivity S%∗i as a function of for input parameter vector
with X0∗∗ ; C0∗∗ ; Ki∗∗ as parameters.

Fig. 8. Normalized objective sensitivity S%∗i as a function of for input parameter vector
with X0∗∗ ; C0∗∗ ; Ki∗∗ as parameters.

Greek letters  speciEc growth rate of biomass, ks−1

= (X ∗∗ − X0∗∗ )=X0∗∗  t=tmax , dimensionless time
= (C0∗∗ − C ∗∗ )=C0∗∗ lag = tlag =tmax ; dimensionless lag time
= pH=pH0 , normalized pH C02
  (=
(1 + (2C0 =Ki ))C0 (KS + C0 + (C02 =Ki ))Ki
'= f+
KS + C0 + (C02 =Ki ) = =, dimensionless group, analogous to the
 parameter used in Eq. (6), Semenov number
(mg dm−3 )−f (ks)−1 c critical value of
5110 S. Dutta et al. / Chemical Engineering Science 56 (2001) 5103–5110
Fig. 9. Normalized objective sensitivity S%∗i as a function of
Subscripts and superscripts
0 initial condition
max maximum value
∗ quantity at min
Acknowledgements
The valuable suggestions given by the reviewers are
gratefully acknowledged.
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