Chemical Engineering Science 61 (2006) 34633475

www.elsevier.com/locate/ces
Modeling of batch phenol biodegradation in internal loop airlift bioreactor
with gas recirculation by Candida tropicalis
Xiaoqiang Jia, Jianping Wen

, Yan Jiang, Xianling Liu, Wei Feng

Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, P.R. China
Received 12 September 2005; received in revised form 21 November 2005; accepted 19 December 2005
Available online 8 February 2006
Abstract
A mathematical model for simulating the dynamic behaviors of batch phenol biodegradation processes in internal loop airlift bioreactor
(ILAB) with gas recirculation using free cells of the yeast Candida tropicalis was established by coupling the uid dynamic model and
the mass balance model. Based on the coupling arithmetic, the program for evaluating batch phenol biodegradation processes was achieved.
The predicted results of linear liquid velocities, gas holdups, linear gas velocities, cell and phenol concentration proles in the riser and the
downcomer of the ILAB with gas recirculation agreed very well with the corresponding experimental data, and the applicability and reliability
of this proposed model were validated.
2006 Elsevier Ltd. All rights reserved.
Keywords: Phenol biodegradation; Candida tropicalis; ILAB; Modeling
1. Introduction
Being produced in several industries and operations such as
petroleum reneries, gas and coke oven industries, ber glass
units, pharmaceuticals, explosive manufacture, phenolic resin
manufacture, plastic and varnish industries, textiles units mak-
ing use of organic dyes, and smelting and related metallurgical
operations, phenol is one of the most common water pollutants
and considered as highly toxic among the environmentally
concerned substances (Kumar et al., 2005). Therefore, it has
been mandatory to treat these wastes before safe disposal
to the water.
The most common method developed to deal with the
removal of phenol from wastewater is biological treatment
(Buitrn et al., 1998; Caldeira et al., 1999; Fang and Zhou,
1997, Gonzlez et al., 2001 a,b; Holub et al., 2000; Khoury
et al., 1992; Sarfaraz et al., 2004; Sutton and Mishra, 1994),
which is generally preferred to physical or chemical treatment
methods because of lower costs and possibility of complete
mineralization (Jiang et al., 2004). Although the biological

Corresponding author. Tel./fax: +86 22 27890492.

E-mail address: jpwen@tju.edu.cn (J.P. Wen).
0009-2509/$ - see front matter 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.ces.2005.12.025
treatment of phenolic wastewater can be achieved with con-
ventional activated sludge systems, such systems have been
known to break down because of uctuations in phenol loads
or exposure to high phenol loading (Watanabe et al., 1999).
Thus, various pure cultures of microbial strains have been
tested for degrading phenolic compounds (Ha and Vinitnan-
tharat, 2000; Li et al., 1998; Santos and Linardi, 2004; Vinit-
nantharat et al., 2001). Among these pure cultures, Jiang et al.
(2005) isolated a strain of yeast Candida tropicalis and showed
that it could degrade very high concentration phenol up
to 2000 mg/L.
Recently, the treatment of wastewater containing phenol fo-
cuses on employing and exploring new type of bioreactors with
high performance for practical utilization. Internal loop airlift
bioreactor (ILAB), characterized by many advantages such as
better mixing, intimate contact between phases, faster oxygen
transfer rate, higher operational exibility, shorter reaction
time and greater processing capability (Wen et al., 2005b), was
preferred to be used in phenol removal to conventional type
of reactors. Raja Rao et al. (1997) found that the biodegrada-
tion of phenol was faster in airlift bioreactor than in bubble
column. Quan et al. (2004) developed an airlift inner-loop
bioreactor packed with honeycomb-like ceramic as the carrier
to immobilize microorganism for simultaneous phenol and
3464 X. Jia et al. / Chemical Engineering Science 61 (2006) 34633475
2, 4-dichlorophenol biodegradation under fed-batch and con-
tinuous operations. Their results showed that both phenol and
2, 4-dichlorophenol could be efciently degraded.
However, studies on the modeling of phenol biodegrada-
tion processes in ILAB are very limited. Livingston and Chase
(1989) proposed a mathematical model to describe simultane-
ous diffusion and reaction of oxygen and phenol in the draft
tube uidized bed reactor (i.e., ILAB). They used this model
to discuss the transition from phenol-limited to oxygen-limited
kinetics of phenol biodegradation. Though this model regarded
the ILAB as a fully-mixed reactor, it did not take into account
the variations of the dynamic behaviors in different zones of
the ILAB.
The aim of this study is to perform hydrodynamic charac-
teristics investigations and batch phenol biodegradation exper-
iments in an ILAB with gas recirculation; then to develop a
mass balance model to simulate the dynamic behaviors of batch
phenol biodegradation processes in different sections of the
ILAB with gas recirculation, coupling a uid dynamic model
to predict the hydrodynamic characteristics; and to validate the
model predictions of the hydrodynamic characteristics and sim-
ulations of phenol biodegradation and cell growth behaviors
using the experimental data.
2. Mathematical model
Based on the following assumptions, a mathematical model
for simulating the dynamic behaviors of batch phenol biodegra-
dation processes in the ILAB with gas recirculation using free
cells of the yeast C. tropicalis was established by coupling the
mass balance model and the uid dynamic model.
(1) The ILAB consists of four sections, i.e., the riser, the
downcomer, the top section and the bottom section. The hydro-
dynamic properties in each section are different, and they are
uniform throughout that section.
(2) The ow pattern of the liquid phase in the riser and the
downcomer can be expressed by one-dimensional axial disper-
sion model, deviation from plug ow because of axial mix-
ing at each circulation and back mixing due to the recycling
(Gavrilescu and Tudose, 1999). The top and the bottom sec-
tions are CSTRs.
(3) Substrate biodegradation and cell growth are only limited
by phenol in the free cells system while all other nutrients
including oxygen are in excess.
(4) Phenol and cells are distributed uniformly throughout
each section initially.
(5) Fluid ow and bioreaction are coupled and interact
with each other. Fluid ow affects the bioreaction, leading
to the change of the liquid physical properties of the fer-
mentation broth, which in turn will affect the hydrodynamic
characteristics.
2.1. Fluid dynamic model
The uid dynamic model for gasliquid two-phase ILAB
with gas recirculation is based on the drift-ux model proposed
by Zuber and Findlay (1965) to estimate the gas holdups and the
linear gas and liquid velocities in the riser and the downcomer.
The linear gas velocities in the riser and the downcomer can
be expressed as
V
gr
= c
r
(U
gr
+ U
lr
) + U
bt ,r
, (1)
V
gd
= c
d
(U
gd
+ U
ld
) U
bt ,d
, (2)
where V
gr
and V
gd
are the linear gas velocities in the riser
and the downcomer, c
r
and c
d
the dimensionless distribution
parameters in the riser and the downcomer, U
gr
and U
gd
the
supercial gas velocities in the riser and the downcomer, U
lr
and U
ld
the supercial liquid velocities in the riser and the
downcomer, U
bt ,r
and U
bt ,d
the bubble terminal velocities in
the riser and the downcomer, respectively.
The gas holdups in the riser and the downcomer can be re-
lated to the supercial gas velocities and the linear gas veloci-
ties in the riser and the downcomer, respectively:

gr
=
U
gr
V
gr
, (3)

gd
=
U
gd
V
gd
, (4)
where
gr
and
gd
are the gas holdups in the riser and the
downcomer, respectively.
Similarly, the relationships among the linear liquid velocities,
supercial liquid velocities and liquid holdups in the riser and
the downcomer are

lr
=
U
lr
V
lr
, (5)

ld
=
U
ld
V
ld
, (6)
where V
lr
and V
ld
are the linear liquid velocities in the riser
and the downcomer,
lr
and
ld
the liquid holdups in the riser
and the downcomer, respectively. Also

lr
= 1
gr
, (7)

ld
= 1
gd
. (8)
The liquid volumetric ow rate through the riser is equal to
that through the downcomer:
U
lr
A
r
= U
ld
A
d
, (9)
where A
r
and A
d
are the cross-sectional areas of the riser and
the downcomer, respectively.
Similarly, a gas-phase volume balance at the riser bottom
depicts that the riser gas volumetric ow rate equates the sum
of the downcomer gas volumetric ow rate and the injection
gas volumetric ow rate, Q
g
:
U
gr
A
r
= U
gd
A
d
+ Q
g
. (10)
Lu and Hwang (1995) found that a linear relationship be-
tween the gas holdups in the riser and the downcomer exists
for gasliquid two-phase ILAB. As the ratio of cross-sectional
X. Jia et al. / Chemical Engineering Science 61 (2006) 34633475 3465
area of the downcomer to the riser of their ILAB is the same
as in this study, Eq. (11) is therefore used in this study for
gasliquid two-phase system:

gd
= 0.8
gr
. (11)
Liquid recirculation in the ILAB is caused by the hydrostatic
pressure difference, P
h
, between the riser and the downcomer
and can be written as (Miyahara and Kawate, 1993):
P
h
= [(
ld

ld
+
g

gd
) (
lr

lr
+
g

gr
)]gH
d
, (12)
where H
d
is the draught tube height.
Moreover, the total frictional loss in the ILAB, P
loss
, can
be calculated as the sum of the frictional losses in the four
sections of the ILAB (Vunjak-Novakovic et al., 1992):
P
loss
= (P
f
)
r
+ (P
f
)
d
+ (P
f
)
t
+ (P
f
)
b
,
(13)
where (P
f
)
r
and (P
f
)
d
are the frictional losses in the
riser and the downcomer, respectively, and they are obtained
by (Verlaan et al., 1986):
(P
f
)
r
=
1
2

lr
k
f r
V
2
lr
, (14)
(P
f
)
d
=
1
2

ld
k
f d
V
2
ld
, (15)
where k
f r
and k
f d
are the frictional loss coefcients in the riser
and the downcomer, respectively.
(P
f
)
t
and (P
f
)
b
are the frictional losses due to ow
reversal at the top and the bottom sections of the ILAB, respec-
tively. They can be obtained by (Livingston and Zhang, 1993)
(P
f
)
t
=
1
2

lt
k
f t
V
2
lr
, (16)
(P
f
)
b
=
1
2

lb
k
f b
V
2
ld
, (17)
where k
f t
and k
f b
are the frictional loss coefcients in the top
and the bottom sections, respectively.
At steady state P
h
=P
loss
(Verlaan et al., 1986), namely:
[(
ld

ld
+
g

gd
) (
lr

lr
+
g

gr
)]gH
d
=
1
2
U
2
ld

(
lt
k
f t
+
lr
k
f r
)
(1
gr
)
2

A
d
A
r

2
+
(
lb
k
f b
+
ld
k
f d
)
(1
gd
)
2

. (18)
2.2. Mass balance model
The mass balance equation for phenol in the riser of the
ILAB is expressed by one-dimensional axial dispersion model:
jS
r
jt
= D
zr

j
2
S
r
jz
2

V
lr

jS
r
jz

S
r
, (19)
while for cells, we have
jX
r
jt
= D
zr

j
2
X
r
jz
2

V
lr

jX
r
jz

+
X
r
. (20)
The mass balance equation for phenol in the downcomer is
jS
d
jt
= D
zd

j
2
S
d
jz
2

+ V
ld

jS
d
jz

S
d
, (21)
while for cells, we have
jX
d
jt
= D
zd

j
2
X
d
jz
2

+ V
ld

jX
d
jz

+
X
d
, (22)
where D
zr
and D
zd
are the respective axial dispersion coef-
cient in the riser and the downcomer, which species the extent
of axial mixing,
S
r
and
S
d
the respective phenol degradation
rate in the riser and the downcomer,
X
r
and
X
d
the cell growth
rate in the riser and the downcomer. Since the direction of ow
in the downcomer is opposite to that in the riser, V
ld
is positive.
Note that the unit of time is in hour.
The mass balance equation for phenol in the top section is
jS
t
jt
=
U
lr
A
r
(S
r
|
z=H
d
S
t
)
v
t

lt

S
t
, (23)
while for cells, we have
jX
t
jt
=
U
lr
A
r
(X
r
|
z=H
d
X
t
)
v
t

lt
+
X
t
. (24)
The mass balance equation for phenol in the bottom section
is
jS
b
jt
=
U
ld
A
d
(S
d
|
z=0
S
b
)
v
b

lb

S
b
, (25)
while for cells, we have
jX
b
jt
=
U
ld
A
d
(X
d
|
z=0
X
b
)
v
b

lb
+
X
b
, (26)
where v
t
and v
b
are the respective volume of the top section
and the bottom section,
S
t
and
S
b
the respective phenol degra-
dation rate in the top section and the bottom section,
X
t
and

X
b
the respective cell growth rate in the top section and the
bottom section.
lt
and
lb
are the respective liquid holdup in
the top section and the bottom section, which are considered to
be the same as the overall liquid holdup of the whole reactor
and calculated as (Chisti, 1989):

lt
=
lb
=
l
= 1
g
= 1

gr
A
r
+
gd
A
d
A
r
+ A
d
. (27)
The initial conditions are
S
r
|
z=z
= S
0
r
, (28)
S
d
|
z=z
= S
0
d
, (29)
S
t
= S
0
t
, (30)
S
b
= S
0
b
, (31)
X
r
|
z=z
= X
0
r
, (32)
X
d
|
z=z
= X
0
d
, (33)
3466 X. Jia et al. / Chemical Engineering Science 61 (2006) 34633475
X
t
= X
0
t
, (34)
X
b
= X
0
b
. (35)
The boundary conditions are
S
r
|
t
z=0
= S
b
|
t
, (36)
jS
r
jz

t
z=H
d
= 0, (37)
S
d
|
t
z=H
d
= S
t
|
t
, (38)
jS
d
jz

t
z=0
= 0, (39)
X
r
|
t
z=0
= X
b
|
t
, (40)
jX
r
jz

t
z=H
d
= 0, (41)
X
d
|
t
z=H
d
= X
t
|
t
, (42)
jX
d
jz

t
z=0
= 0. (43)
2.3. Model parameters
2.3.1. Distribution parameter c
i
The distribution parameter, c
i
, ranges between 1.02 and 1.17
(Zuber and Findlay, 1965). Since the ow pattern in our system
is similar to that in the internal loop airlift system of Hatch
(1973), the value of c
i
(=c
r
= c
d
) used in this study is 1.065.
2.3.2. Bubble terminal velocity U
bt ,i
The bubble terminal velocity, U
bt ,i
, is calculated using the
equation proposed by Wilkinson (1981) for non-coalescent sys-
tem:
U
bt ,i

l,m

l,m
= 2.25

3
l,m

l,m
g
4
l,m

0.273

l,m

0.03
. (44)
For coalescent system, the bubble terminal velocity is given
by (Hwang and Cheng, 1997)
U
bt ,i
= 1.0(gd
bi
)
0.5
, (45)
where the bubble diameter, d
bi
, for coalescent system is calcu-
lated as (Bhavaraju et al., 1978)
d
bi
= 0.7

0.6
li

P
gi
v
li

0.4

0.2
li

li

, (46)
where (P
gi
/v
li
) is the specic power input obtained by the
following equation (Chisti, 1989):
P
gi
v
li
=

li
gU
gr
1 + (A
d
/A
r
)
. (47)
The liquid phase density,
li
, viscosity,
li
, and surface ten-
sion,
li
, in the riser and the downcomer are related to the resid-
ual phenol and cell concentrations. Under the condition that the
initial phenol concentration varied from 100 to 2000 mg/L and
the initial cell concentration varied from 10 to 100 mg/L, the
following expressions can be obtained (Cao and Wen, 2005):

li

l,m
= 1 + 1 10
6
S
i
[S]
4 10
6
X
i
[X]
(R
2
= 0.972), (48)

li

l,m
= 1 4 10
5
S
i
[S]
9 10
9

S
i
[S]

2
+ 1 10
3
X
i
[X]
8 10
7

X
i
[X]

2
(R
2
= 0.963), (49)

li

l,m
= 1 1 10
5
S
i
[S]
1 10
8

S
i
[S]

2
+ 5 10
12

S
i
[S]

3
1.2 10
3
X
i
[X]
+ 5 10
5

X
i
[X]

2
8 10
8

X
i
[X]

3
(R
2
= 0.971), (50)
where
l,m
,
l,m
and
l,m
are the liquid density, liquid viscos-
ity and liquid surface tension of the mineral salt medium and
were measured initially experimentally; S
i
and X
i
are the con-
centrations of phenol and cells respectively, in the riser and the
downcomer, [S] and [X] are the unit concentrations of phenol
and cells, respectively.
2.3.3. Frictional loss coefcients, k
f r
and k
f d
, in the riser
and the downcomer
The frictional loss coefcients, k
f r
and k
f d
, are calculated
using the following equations:
k
f r
= 4f
r
H
d
D
r
, (51)
k
f d
= 4f
d
H
d
D
d
, (52)
where D
r
and D
d
are the respective diameter of the riser and
the downcomer, f
r
and f
d
the frictional factors of the liquid
phase in the riser and the downcomer, respectively, and they
are estimated from the Blasius equation:
f
r
= 0.0791 Re
0.25
lr
, (53)
f
d
= 0.0791 Re
0.25
ld
, (54)
where Reynolds numbers of the liquid phase in the riser and
the downcomer are
Re
lr
=

lr
V
lr
D
r

lr
, (55)
Re
ld
=

ld
V
ld
D
d

ld
. (56)
X. Jia et al. / Chemical Engineering Science 61 (2006) 34633475 3467
Fig. 1. Schematic diagram describing the modeling of uid dynamics and mass balance.
3468 X. Jia et al. / Chemical Engineering Science 61 (2006) 34633475
2.3.4. Frictional loss coefcients, k
f t
and k
f b
, in the top
section and the bottom section
The method developed by Chisti et al. (1988) is used in this
study to estimate k
f t
and k
f b
. They observed that k
f t
k
f b
for the ILAB, thus k
f t
is negligible. The correlation for k
f b
is
k
f b
= 11.402((A
d
/A
b
))
0.789
. (57)
This correlation is valid for an (A
d
/A
b
) range of 0.21.8.
A
b
is the free area below the draught tube.
2.3.5. Axial dispersion coefcients
Axial dispersion coefcients can be derived from empirical
correlations. In this study, the correlation proposed by Znad
et al. (2004) for ILAB is adopted:
D
zr
= 2.61D
1.5
r
U
0.5
gr
, (58)
D
zd
= 2.61D
1.5
d
U
0.5
gd
. (59)
Thus both the axial dispersion coefcients in the riser and the
downcomer increase with an increase in gas volumetric ow
rate.
2.3.6. Intrinsic kinetic parameters
The intrinsic cell growth and phenol biodegradation kinetics
of the yeast C. tropicalis were determined experimentally in
shaking asks (Jiang et al., 2005):

X
=
X
X =
dX
dt
=

max
S
K
s
+ S + S
2
/K
i
X
=
0.48S
11.7 + S + (S
2
/207.9)
X (R
2
= 0.996), (60)

S
=
S
X =
dS
dt
=
X
+ X
= 0.823
X
+ 0.277X (R
2
= 0.994). (61)
2.4. Model solution
The proposed model for simulating the dynamic behaviors
of batch phenol biodegradation processes was solved by the
coupling arithmetic. The uid dynamic model was solved us-
ing the Mathematica Software v5.0 with a relative tolerance
(RT) of 1 10
2
for the calculating of the linear liquid veloc-
ity in the dowmcomer, V
ld
, and the mass balance model was
solved by the nite element analysis method using the soft-
ware package of Femlab 3.0 a with a time step length (TSL) of
0.01 h and a RT of 1 10
2
for the calculation of each phenol
and cell concentration in the four sections of the ILAB. Fig. 1
is the schematic diagram describing the modeling of the mass
balance and uid dynamics. A typical solver ran 34 h of com-
puting time took about 10 h of wall clock time for the evalu-
ation of this proposed model, as most time was spent on the
coupling of the uid dynamics and the mass balance. In the
absence of the initial biomass and phenol concentrations, this
model for predicting the hydrodynamic characteristics of the
gasliquid two-phase ILAB without bioreaction was solved us-
ing the Mathematica Software v5.0 with a RT of 1 10
2
for
the calculation of the linear liquid velocity in the dowmcomer,
V
ld
. A typical solver took less than 2 s of wall clock time for
evaluation of the uid dynamics only.
3. Experimental procedure
3.1. Bioreactor
The ILAB was constructed of 0.12 m ID Plexiglas column
1.5 min height. The draft tube was 0.8 min height and was xed
0.1 m above the bottom of the bioreactor. The inside diameter of
the draft tube was 0.08 m(wall thickness 0.001 m). The distance
between the liquid surface free of air and the upper end of the
draft tube was 0.10 m. The diameter of the concentric jet nozzle
was 0.0054 m, and located at 0.075 m above the bottom of the
bioreactor. The gas volumetric ow rate was controlled by a
rotameter, and was varied from 0.3 10
3
1.6 10
3
m
3
/s,
corresponding to linear liquid velocity larger than the bubble
terminal velocity, which ensures the ILAB to be operated under
the gas recirculation ow regime (van Benthum et al., 1999).
Sterilized air was provided by passing air through an air lter
column, and there was an additional air lter above the top
of the bioreactor to prevent other bacterial contamination as
shown in Fig. 2.
3.2. Hydrodynamic characteristics investigations
In order to eliminate the disturbance of fermentation broth
on the measuring electrodes and to enhance the measurement
veracity, the mineral salt medium-air system without bioreac-
tion was used for measuring the hydrodynamic characteristics.
The liquid mineral salt medium contained (g/L): 0.4 K
2
HPO
4
,
0.2 KH
2
PO
4
, 0.1 NaCl, 0.1 MgSO
4
, 0.01 MnSO
4
H
2
O, 0.01
Fe
2
(SO
4
)
3
H
2
O, 0.01 Na
2
MoO
4
2H
2
O, 0.4 (NH
4
)
2
SO
4
(pH = 6.0).
The linear liquid velocities in the riser and the downcomer
were obtained by the tracer response method; the pH electrodes
were located at 0.2 and 0.8 mabove the bottomof the bioreactor,
respectively (Lu et al., 1994; Wen et al., 2005a). Gas holdups
and linear gas velocities in the riser and the downcomer were
obtained by ve-point conductivity probe located at 0.4 mabove
the bottom of the bioreactor, respectively (Wen et al., 2002).
3.3. Batch phenol biodegradation experiments
Pure culture of the yeast C. tropicalis stored in our lab was
used for batch phenol biodegradation in the ILAB in this study.
This strain was isolated with the ability to utilize phenol as sole
carbon source and could tolerate very high phenol concentra-
tion up to 2000 mg/L (Jiang et al., 2005). Liquid cultures were
grown in the mineral salt medium supplemented with 500 mg/L
phenol at 30

C in a rotary shaker at 200 rpm. Phenol was lter-

sterilized through membranes (pore size of 0.2 m) and added
to the medium before inoculation (Kibret et al., 2000; Lonard
and Lindley, 1999).
X. Jia et al. / Chemical Engineering Science 61 (2006) 34633475 3469
Fig. 2. Schematic diagram of the experimental apparatus.
Fig. 3. Comparison between model predicted and experimental measured
linear liquid velocity variations with gas volumetric ow rate in the riser and
the downcomer.
For each batch phenol biodegradation experiment, after
washing with sterilized deionized water and sterilized mineral
salt medium, 11.3 L mineral salt medium containing the initial
phenol concentration of 2000 mg/L and cell concentration of
50 mg/L was added into the bioreactor. All experiments were
Fig. 4. Comparison between model predicted and experimental measured gas
holdup variations with gas volumetric ow rate in the riser and the downcomer.
carried out at the initial pH of 6.0. The temperature was kept
at 30

C by circulating thermostated water in the water jacket.

To measure the residual phenol concentration and cell con-
centration in the riser and the downcomer, there were four holes
on each side along the height of the ILAB for periodically
sampling the fermentation broth: 0.2, 0.4, 0.6 and 0.8 m above
3470 X. Jia et al. / Chemical Engineering Science 61 (2006) 34633475
the bottom of the bioreactor. The residual phenol concentra-
tion was measured using high-performance liquid chromatog-
raphy (HPLC, model Series III, LabAlliance, USA) equipped
with a C18 column (250 4.6 mm, LabAlliance, USA). The
absorbance of the efuent solution was continuously measured
Fig. 5. Comparison between model predicted and experimental measured
linear gas velocity variations with gas volumetric ow rate in the riser and
the downcomer.
Fig. 6. Comparison between model simulated and experimental measured phenol concentration proles with time and axial course in the riser and downcomer
at xed gas volumetric rate Q
g
= 1 10
3
m
3
/s, xed initial cell concentration (X
0
= 50 mg/L) and different initial phenol concentrations.
at 280 nm. The ow rate was 1.0 mL/min and the mobile
phase was 57.1% (V/V) of methanol. The culture broth was
centrifuged at 7500 rpm for 10 min, and the supernatant was
analyzed. Cell concentration was monitored spectrophoto-
metrically by measuring the absorbance at the wavelength of
600 nm. Cell concentration on a dry weight basis was mea-
sured by ltering the cell suspension with a lter and drying
the lter paper and cells to a constant weight for 24 h at
105

C. A linear relationship between dry cells weight and

the optical density (OD) was obtained: cell concentration
(mg/L) = 346.78 OD
600
0.6923.
All the experiments were performed in triplicate and the data
shown in the corresponding gures of the following section are
the mean values of the experiments.
4. Model validation
The proposed mathematical model can simulate the hy-
drodynamic behaviors and biodegradation behaviors of batch
phenol biodegradation processes in ILAB with gas recircula-
tion by coupling the uid dynamic model and the mass balance
model. It is very perfect for the dynamic parameters of the
hydrodynamic and phenol biodegradation characteristics to be
utilized simultaneously for validating the proposed mathemat-
ical model. However, the fermentation broth will disturb the
X. Jia et al. / Chemical Engineering Science 61 (2006) 34633475 3471
Fig. 7. Comparison between model simulated and experimental measured cell concentration proles with time and axial course in the riser and downcomer at
xed gas volumetric rate Q
g
= 1 10
3
m
3
/s, xed initial cell concentration (X
0
= 50 mg/L) and different initial phenol concentrations.
measuring electrodes thereby affecting the measurement ve-
racity of the hydrodynamic characteristics. In order to effec-
tively evaluate the proposed mathematical model, the validation
of hydrodynamic characteristics and the validation of phenol
biodegradation behaviors are adopted respectively. Firstly, un-
der the condition that cells and phenol were not added into the
mineral salt medium, the proposed mathematical model was
simplied to the uid dynamic model only; and the measured
linear liquid velocities, gas holdups and linear gas velocities
in the riser and the downcomer were used to validate the hy-
drodynamic characteristics predicted by this proposed mathe-
matical model. Secondly, in the actual phenol biodegradation
processes, the experimentally measured phenol and cell con-
centration proles were adopted for validating the dynamic be-
haviors of batch phenol biodegradation processes simulated by
this proposed mathematical model.
4.1. Validation of the uid dynamic model
Fig. 3 shows the comparison between model predictions and
experimental measurements of the variations of the linear liquid
velocities with gas volumetric ow rate in the riser and the
downcomer of the ILAB with gas recirculation. It can be seen
that the linear liquid velocity in the riser is larger than that in
the downcomer, both of which increase with the increase in
gas volumetric ow rate. It also can be seen that the model
simulations agree exceedingly well with the experimental data.
Fig. 4 compares model-simulated results and experimental
data of the variations of gas holdups with gas volumetric ow
rate in the riser and the downcomer of the ILAB with gas re-
circulation. Excellent agreement is achieved between the pre-
dicted results and experimental data. It is noted that the gas
holdup in the riser is larger than that in the downcomer and both
of them increase with increase in the gas volumetric ow rate.
Fig. 5 is the comparison between model predictions and
experimental measurements of the variations of the linear gas
velocities with gas volumetric ow rates in the riser and the
downcomer of the ILAB with gas recirculation. Excellent
agreement is achieved between the predicted results and exper-
imental measurements. It is obvious that the linear gas velocity
in the riser is much larger than that in the downcomer, both of
which increase with the increase in gas volumetric ow rate.
In addition, the linear gas velocities in the downcomer are all
positive under the gas volumetric ow rates investigated, in-
dicating that the ILAB is operated under the gas recirculation
ow regime.
From Figs. 35, it can be observed that this proposed
mathematical model can be convincingly used to predict the
3472 X. Jia et al. / Chemical Engineering Science 61 (2006) 34633475
Fig. 8. Comparison between model simulated and experimental measured phenol concentration proles with time course in the riser and downcomer at xed
axial position z = 0.6 m, xed initial phenol concentration (S
0
= 2000 mg/L) and different gas volumetric ow rates, different initial cell concentrations.
hydrodynamic characteristics of the ILAB in the absence of
bioreaction, such as linear liquid velocities, gas holdups and
linear gas velocities in the riser and the downcomer. And the
relative standard deviations of the experimental data in each
gure are all within 3%, which ensure the reliability of the
experimental results. Additionally, the comparisons between
the model simulations and experimental data in our study to
the model predictions by Lu and Hwang (1995) have also been
included in Figs. 35. Both of the two models can convincingly
predict the hydrodynamic characteristics. And the two models
themselves are in good accordance except for some minor
differences, which may be the reason that we have adopted
different correlations to calculate the bubble terminal velocity.
4.2. Validation of phenol biodegradation behaviors
At the xed gas volumetric ow rate of 1 10
3
m
3
/s and
initial cell concentration of 50 mg/L, the changes of the sim-
ulated and measured residual phenol and cell concentrations
with time and axial position in the riser and the downcomer
of the ILAB with gas recirculation are illustrated in Figs. 6
and 7 at three initial phenol concentrations: 2000, 1800 and
1600 mg/L. It is observed from Fig. 6 that the residual phenol
concentrations in the riser and the downcomer reduce to zero
after 34 h for initial phenol concentration of 2000 mg/L at all
axial positions, which is about 4 and 7 h longer than that for ini-
tial phenol concentration of 1800 and 1600 mg/L, respectively.
The increase in the cell concentration of lower initial phenol
concentration is faster than that of higher initial phenol concen-
tration because of weaker substrate inhibition effect, while the
nal cell concentration of higher initial phenol concentration
is larger than that of lower initial phenol concentration as the
former experiences a longer growth process and more substrate
can be utilized for biomass accumulation. It also can be seen
from these two gures that the uctuations of residual phenol
and cell concentrations at any time along the axial course in the
riser and the downcomer are negligible. And it is interesting to
nd that the phenol and cell concentrations in the riser and in
the downcomer are almost the same, indicating that the ILAB
in the gas recirculation regime has a good mixing property.
Figs. 8 and 9 show the changes of model predicted and ex-
perimentally measured residual phenol and cell concentrations
X. Jia et al. / Chemical Engineering Science 61 (2006) 34633475 3473
Fig. 9. Comparison between model simulated and experimental measured cell concentration proles with time course in the riser and downcomer at xed
axial position z = 0.6 m, xed initial phenol concentration (S
0
= 2000 mg/L) and different gas volumetric ow rates, different initial cell concentrations.
with time and gas volumetric ow rate in the riser and the
downcomer of the ILAB with gas recirculation at the same ax-
ial position of z =0.6 m, the same initial phenol concentration
of 2000 mg/L and different initial cell concentrations of 50, 60
and 70 mg/L. Higher initial cell concentration indicates higher
phenol consumption rate, resulting in shorter degradation pe-
riod as shown in Fig. 8. It is interesting to nd in Fig. 9 that
cell concentration of higher initial cell concentration is larger
than that of lower initial cell concentration at any given time,
but the nal cell concentration of the former is a little smaller,
which may be the reason that more substrate is consumed for
biomass maintenance. And it can be concluded that an increase
in the gas volumetric ow rate has no signicant effects on
the phenol and cell concentration proles in the riser and the
downcomer, as long as the ow pattern of the ILAB reaches to
complete gas recirculation regime.
From Figs. 6 to 9, it is clear that the agreements between
the simulated results and corresponding experimental data of
phenol and cell concentration proles in the riser and the down-
comer of the ILAB with gas recirculation are very good. It is in-
dicated that this proposed mathematical model can be adopted
for simulating the dynamic behaviors of batch phenol biodegra-
dation processes using free cells of the yeast C. tropicalis in
the ILAB with gas recirculation. And the relative standard de-
viations in each gure are all within 3%, which ensures the
reliability of the experimental data.
5. Conclusion
In this work, a mathematical model was proposed by cou-
pling the uid dynamic model and the mass balance model to
evaluate the dynamic behaviors of batch phenol biodegrada-
tion processes in ILAB with gas recirculation using free cells
of the yeast C. tropicalis, taking into account uid ow, ax-
ial dispersion and bioreaction. The validation of the uid dy-
namics was performed in mineral salt medium-air two-phase
system in the absence of the initial biomass and phenol con-
centrations. The model predicted hydrodynamic characteristics
such as linear liquid velocities, gas holdups and linear gas
velocities in the riser and the downcomer of the ILAB with
gas recirculation agreed very well with the corresponding ex-
perimental data and also the model simulations proposed by
3474 X. Jia et al. / Chemical Engineering Science 61 (2006) 34633475
Lu and Hwang (1995). The validation of the dynamic behaviors
of batch phenol biodegradation processes was carried out in
actual phenol biodegradation processes in terms of phenol and
cell concentration proles in the riser and downcomer of the
ILAB. The model simulations were in good consistency with
the experimental data. Thus the proposed model can provide
valuable directions for reactor design, optimization and scale-
up in full-scale applications, which will signicantly reduce the
cost of unnecessary experimental work.
In addition, the nal evaluation standard of the phenol
biodegradation processes is also presented as the removal ef-
ciencies of TOC, COD or BOD, besides the residual phenol
concentration. The combination of this proposed model with
intrinsic biodegradation kinetics of TOC, COD or BOD can be
applied to predict the biodegradation behaviors and distribution
in different zones of the ILAB not only for phenol, but also
for TOC, COD or BOD. This is very valuable and important
information in full-scale applications of phenolic wastewater
treatment processes.
Notation
a constant, dimensionless
A cross-sectional area, m
2
A
b
the free area below the draught tube, m
2
b constant, dimensionless
BOD biological oxygen demand, mg/L
c distribution parameter, dimensionless
COD chemical oxygen demand, mg/L
d
b
average bubble diameter, m
D diameter, m
D
z
axial dispersion coefcient, m
2
/s
f frictional factor, dimensionless
H
d
draught tube height, m
k
f
friction coefcient, dimensionless
K
i
inhibition coefcient, mg/L
K
s
saturation constant, mg/L
P
f
frictional pressure drop, Pa
P
g
power input, Pa
P
h
hydrostatic pressure difference between the riser
and the downcomer, Pa
P
loss
total frictional loss, Pa
Q volumetric ow rate, m
3
/s
R
2
correlation coefcient, dimensionless
Re Reynolds number, dimensionless
S phenol concentration, mg/L
[S] unit concentration of phenol, 1 mg/L
t time, h
TOC total organic carbon, mg/L
U supercial velocity, m/s
U
bt
bubble terminal velocity, m/s
v volume, m
3
V linear velocity, m/s
X cell concentration, mg/L
[X] concentration of cells, 1 mg/L
z axial position, m
Greek letters
growth associated constant for phenol consump-
tion, dimensionless
non-growth associated constant for phenol con-
sumption, h
1
rate of reaction, g/Lh
holdup, dimensionless
specic rate, h
1

l
liquid viscosity, Pa s

max
maximum specic growth rate, h
1
density, kg/m
3
surface tension, N/m
Superscripts
0 initial condition
t time
Subscripts
b bottom section
c column
d downcomer
g gas phase
i riser or downcomer
l liquid phase
m mineral salt medium
r riser
S phenol
t top section
X cell
Acknowledgement
The authors wish to acknowledge the nancial support pro-
vided by the Key National Natural Science Foundation of China
(No. 20336030), Key Natural Science Foundation of Tianjin
(No. 05YFJZJC 00500), Program for New Century Excellent
Talents in University and Program for Changjiang Scholars and
Innovative Research Team in University.
References
Bhavaraju, S.M., Russell, T.W.F., Blance, H.W., 1978. The design of gas
sparged devices for viscous liquid systems. A.I.Ch.E. Journal 24, 454466.
Buitrn, G., Gonzlez, A., Lpez-Marin, L.M., 1998. Biodegradation of
phenolic compounds by an acclimated activated sludge and isolated
bacteria. Water Science and Technology 37, 371378.
Caldeira, M., Heald, S.C., Carvalho, M.F., Vasconcelos, I., Bull, A.T., Castro,
P.M.L., 1999. 4-Chlorophenol degradation by a bacterial consortium:
development of a granular activated carbon biolm reactor. Applied
Microbiology and Biotechnology 52, 722729.
Cao, J.Y., Wen, J.P., 2005. Fluid hydrodanymics and physical properties
of phenol biodegradation in bioreactors. Bachelor Dissertation, Tianjin
University.
Chisti, M.Y., 1989. Airlift Bioreactors. Elsevier, New York.
Chisti, M.Y., Halard, B., Moo-Yong, M., 1988. Liquid circulation in airlift
reactors. Chemical Engineering Science 43, 451457.
X. Jia et al. / Chemical Engineering Science 61 (2006) 34633475 3475
Fang, H.H.P., Zhou, G.M., 1997. Denitrication of phenolic wastewater by
immobilized sludge. Environmental Technology 18, 827834.
Gavrilescu, M., Tudose, R.Z., 1999. Modelling mixing parameters in
concentric-tube airlift bioreactors: Part II. Axial dispersion. Bioprocess
Engineering 20, 491497.
Gonzlez, G., Herrera, M.G., Garca, M.T., Pea, M.M., 2001a.
Biodegradation of phenol in a continuous process: comparative study of
stirred tank and uidized-bed bioreactors. Bioresource Technology 76,
245251.
Gonzlez, G., Herrera, G., Garca, M.T., Pea, M.M., 2001b. Biodegradation
of phenolic industrial wastewater in a uidized bed bioreactor with
immobilized cells of Pseudomonas putida. Bioresource Technology 80,
137142.
Ha, S.R., Vinitnantharat, S., 2000. Competitive removal of phenol and 2,
4-dichlorophenol in biological activated carbon system. Environmental
Technology 21, 387396.
Hatch, R.T., 1973. Experimental and theoretical studies of oxygen transfer
in the airlift fermentor. Ph.D. Thesis, MIT, Cambridge.
Holub, W., Przytocka-Jusiak, M., Blaszczyk, M., Mycielski, R., 2000. Nitrite
as agent selecting anaerobic phenol-degrading microora in petroleum
rening sediments. Water Research 34, 13541358.
Hwang, S.J., Cheng, Y.L., 1997. Gas holdup and liquid velocity in three-
phase internal-loop airlift reactors. Chemical Engineering Science 52,
39493960.
Jiang, H.L., Tay, J.H., Maszenan, A.M., Tay, S.T.L., 2004. Bacterial diversity
and function of aerobic granules engineered in a sequencing batch reactor
for phenol degradation. Applied and Environmental Microbiology 70,
67676775.
Jiang, Y., Wen, J.P., Li, H.M., Yang, S.L., 2005. The biodegradation of phenol
at high initial concentration by the yeast Candida tropicalis. Biochemical
Engineering Journal 24, 243247.
Khoury, N., Dott, W., Kampfer, P., 1992. Anaerobic degradation of phenol
in batch and continuous cultures by a denitrifying bacterial consortium.
Applied Microbiology Biotechnology 37, 524528.
Kibret, M., Somitsch, W., Robra, K.H., 2000. Characterization of a phenol
degrading mixed population by enzyme assay. Water Research 34,
11271134.
Kumar, A., Kumar, S., Kumar, S., 2005. Biodegradation kinetics of phenol and
catechol using Pseudomonas putida MTCC 1194. Biochemical Engineering
Journal 22, 151159.
Lonard, D., Lindley, N.D., 1999. Growth of Ralstonia eutropha on
inhibitory concentrations of phenol: diminished growth can be attributed
to hydrophobic perturbation of phenol hydroxylase activity. Enzyme and
Microbial Technology 25, 271277.
Li, P.H.Y., Roddick, F.A., Hobday, M.D., 1998. Bioregeneration involving a
coal-based adsorbent used for removing nitrophenol from water. Journal
of Chemical Technology and Biotechnology 73, 405413.
Livingston, A.G., Chase, H.A., 1989. Modeling phenol degradation in a
uidized-bed bioreactor. A.I.Ch.E. Journal 35, 19801992.
Livingston, A.G., Zhang, S.F., 1993. Hydrodynamic behaviour of three-
phase (gasliquidsolid) airlift reactors. Chemical Engineering Science 48,
16411654.
Lu, W.J., Hwang, S.J., 1995. Liquid velocity and gas holdup in three-
phase internal loop airlift reactors with low-density particles. Chemical
Engineering Science 50, 13011310.
Lu, W.J., Hwang, S.J., Chang, C.M., 1994. Liquid mixing in two- and three-
phase airlift reactors. Chemical Engineering Science 49, 14651468.
Miyahara, T., Kawate, O., 1993. Hydrodynamics of a solid-suspended bubble
column with a draught tube containing low-density particles. Chemical
Engineering Science 48, 127133.
Quan, X.C., Shi, H., Zhang, Y.M., Wang, J.L., Qian, Y., 2004.
Biodegradation of 2,4-dichlorophenol and phenol in an airlift inner-loop
bioreactor immobilized with Achromobacter sp. Separation and Purication
Technology 34, 97103.
Raja Rao, T.B.M.L., Sonolikar, R.L., Pentu Saheb, S., 1997. Inuence of
magnetic eld on the performance of bubble columns and airlift bioreactor
with submersed microorganisms. Chemical Engineering Science 52,
41554160.
Santos, V.L., Linardi, V.R., 2004. Biodegradation of phenol by a lamentous
fungi isolated from industrial efuents-identication and degradation
potential. Process Biochemistry 39, 10011006.
Sarfaraz, S., Thomas, S., Tewari, U.K., Iyengar, L., 2004. Anoxic treatment
of phenolic wastewater in sequencing batch reactor. Water Research 38,
965971.
Sutton, P.W., Mishra, P.N., 1994. Activated carbon based biological uidized
beds for contaminated water and wastewater treatment: a state-of-the-art
review. Water Science and Technology 29, 309317.
van Benthum, W.A.J., van der Lans, R.G.J.M., van Loosdrecht, M.C.M.,
Heijnen, J.J., 1999. Bubble recirculation regimes in an internal-loop airlift
reactor. Chemical Engineering Science 54, 39954006.
Verlaan, P., Tramper, J., Vant Riet, K., Luyben, K.C.A.M., 1986. A
hydrodynamic model for an airlift-loop bioreactor with external loop.
Chemical Engineering Journal 33, B43B53.
Vinitnantharat, S., Baral, A., Ishibashi, Y., Ha, S.R., 2001. Quantitative
bioregeneration of granular activated carbon loaded with phenol and 2,4-
dichlorophenol. Environmental Technology 22, 339344.
Vunjak-Novakovic, G., Jovanovic, G., Kundakovic, L., Obradovic, B., 1992.
Flow regimes and liquid mixing in a draft tube gasliquidsolid uidized
bed. Chemical Engineering Science 47, 34513458.
Watanabe, K., Teramoto, M., Harayama, S., 1999. An outbreak of
nonocculation catabolic populations caused the breakdown of a phenol-
digesting activated sludge process. Applied Environmental Microbiology
65, 28132819.
Wen, J.P., Zhou, H., Chen, Y.L., 2002. Local gas phase ow characteristics
of gasliquidsolid three-phase reversed ow jet loop reactor. Chinese
Journal of Chemical Engineering 10, 119122.
Wen, J.P., Jia, X.Q., Cheng, X.R., Yang, P., 2005a. Characteristics of three-
phase internal loop airlift bioreactors with complete gas recirculation
for non-Newtonian uids. Bioprocess and Biosystems Engineering 27,
193205.
Wen, J.P., Jia, X.Q., Pan, L., Wang, C.L., Mao, G.Z., 2005b. Nitrifying
treatment of wastewater from fertilizer production in a multiple airlift loop
bioreactor. Biochemical Engineering Journal 25, 3337.
Wilkinson, P.M., 1981. Physical aspects and scale up of high pressure bubble
columns. Ph.D. Thesis, Groningen, Netherlands.
Znad, H., Ble, V., Marko, J., Kawase, Y., 2004. Modeling and simulation
of airlift bioreactors. Biochemical Engineering Journal 21, 7381.
Zuber, N., Findlay, J.A., 1965. Average volumetric concentration in two-phase
ow systems. Journal of Heat Transfer 87, 453468.