Chemical Engineering Science 54 (1999) 3153}3162

Modeling and simulation of a tubular recycle photobioreactor for

macroalgal cell suspension cultures
G.L. Rorrer*, R.K. Mullikin
Department of Chemical Engineering, Oregon State University, Corvallis, OR 97331,USA

Abstract

Photosynthetic cell suspension cultures derived from marine plants have the potential to produce biomass containing pharmacolo-
gically active metabolites within photobioreactor systems under controlled conditions. The tubular recycle photobioreactor in
particular can promote the mass cultivation of photosynthetic cell suspension cultures. This photobioreactor has two sections:
a coiled tubular section which illuminates the culture with a small light path, and a non-illuminated aeration tank which supplies
dissolved CO needed for photosynthetic biomass production. The culture is recycled between the two sections. A batch reactor

model was developed to predict the cell density vs. time pro"le for the cultivation of photosynthetic cell suspension cultures in the
tubular recycle photobioreactor. This model uniquely couples culture illumination parameters in the tubular section to interphase
CO mass transfer parameters in the aeration tank. Model simulations show that the tubular recycle photobioreactor is operating at

saturation growth kinetics with respect to light. The model predicts a critical cell density at which photosynthetic biomass production
switches from a rate-limited process to a CO delivery limited process. Rate-limited growth proceeds only to this critical cell density,

and then further growth is CO mass transfer limited until the "nal cell density at complete dissolved limiting nutrient consumption is

achieved. Sensitivity analyses reveal that the CO mass transfer coe$cient in the aeration tank, the residence time of the culture in the

tubular section, the ratio of tubular section to aeration tank liquid volume, and the partial pressure of CO in the aeration gas all

a!ect the overall biomass production rate by a!ecting the CO delivery modes to the tubular photobioreactor.  1999 Elsevier

Science Ltd. All rights reserved.

Keywords: Algae; Bioreactor; Mass transfer; Photosynthetic; Tubular

1. Introduction marine seaweeds (Qi and Rorrer, 1995; Rorrer et al.,

Macrophytic marine algae (seaweeds) are a unique
source of pharmacologically active compounds (Carte,
1996). Many seaweeds which produce these compounds
are delicate and complex plants sparsely distributed in
fragile marine ecosystems such as tide pools or coral
reefs, and hence are di$cult to cultivate in the natural
environment. However, cell and tissue suspension cul-
tures established from marine seaweeds have the poten-
tial to biosynthesize these compounds in a controlled
environment within bioreactor systems at a scale re-
quired for further bioprocess development or commercial
production. Toward this end, our laboratory has de-
veloped several photosynthetic cell and tissue suspension
culture systems representing all three major divisions of
* Corresponding author. Tel.: 541 737 3370; fax: 541 737 4600;
e-mail: rorrergl@che.orst.edu.
1996; Huang et al., 1998). We also demonstrated their
cultivation in illuminated stirred-tank, bubble-column,
and tubular bioreactors (Qi and Rorrer, 1995; Rorrer et
al., 1996; Zhi and Rorrer, 1996; Mullikin and Rorrer,
1998) and characterized bioactive eicosanoid product
patterns (Rorrer et al., 1997).
Mass cultivation of newly developed macroalgal sus-
pension cultures described above generically share many
of the scale-up concerns with established microalgal sus-
pension cultures. An important reactor design issue
facing all photosynthetic cell suspension cultures is the
delivery of light to the bioreactor and utilization of light
by the culture (Richmond, 1996). Insu$cient light trans-
mission through dense suspension cultures ultimately
reduces or halts the biomass production rate if the critical
path for light transfer is too large. Traditionally, photo-
bioreactor design for microalgal suspension culture has
focused on minimizing the critical path for light transfer
and maximizing the illumination surface area to culture

0009-2509/99/$ } see front matter  1999 Elsevier Science Ltd. All rights reserved.
PII: S 0 0 0 9 - 2 5 0 9 ( 9 8 ) 0 0 2 9 6 - 6
3154 G.L. Rorrer, R.K. Mullikin/Chemical Engineering Science 54 (1999) 3153}3162
volume ratio. This has been accomplished through a
variety of elegant con"gurations, for example the inter-
nally illuminated "ber-optic photobioreactor (Javanmar-
dian and Palsson, 1991), the #at panel photobioreactor
(Pulz et al., 1995), and the tubular photobioreactor (Pirt
et al., 1983). Tubular photobioreactors are particularly
attractive because they are relatively simple to fabricate
and scale up. There are many variations on the basic
tubular photobioreactor design, for example the a-type
coil con"guration (Lee et al., 1995), the helically wound
coil con"guration (Lee and Bazin, 1990; Watanabe et al.,
1995), and the inclined airlift tubular loop con"guration
(Tredici and Materassi, 1992; Grima et al., 1995). Most
tubular photobioreactor con"gurations share two com-
mon components: a tubular section for culture illumina-
tion, and an aeration section for gas exchange. Often, the
tubular section is not aerated to fully utilize the reactor
volume and to avoid di$culties associated with pneu-
matic gas delivery through the tubing network. Therefore
the aeration section usually carries out the absorption of
CO needed for photosynthetic biomass production and

stripping of dissolved O evolved by photosynthesis.

Carbon dioxide mass transfer in tubular photobioreactor
systems is not well characterized (Borowitzka, 1996).
In our previous work, photosynthetic cell suspension
cultures derived from the macrophytic brown alga
¸aminaria saccharina were successfully cultivated in the
3 l tubular recycle photobioreactor (Mullikin and Rorrer,
1998) shown in Fig. 1. The tubular recycle photobioreac-
tor can be operated in three modes: batch, semi-continu-
ous, and continuous-recycle. A three-way solenoid valve
at the tubular section outlet sets the mode of cultivation.
Fig. 1. 3-l tubular reactor photobioreactor, featuring coiled tubular section, aeration tank, and airlift injection system. In batch operation, the product
outlet line is closed and the culture continually recirculates between the aeration tank and the tubular section.
In batch operation, the solenoid valve is closed, and the
cell suspension culture exiting the tubular section is re-
turned to the aeration tank (total recycle). In semi-con-
tinuous or continuous recycle operation, the solenoid
valve is opened and closed periodically, and a portion of
the culture is drawn o! as product while the remainder is
returned to the tank. Fresh medium is concurrently ad-
ded to the tank while product is removed. ¸aminaria
saccharina cell suspensions have also been cultivated in
externally illuminated, stirred-tank and bubble-column
bioreactors (Qi and Rorrer, 1995; Zhi and Rorrer, 1996).
In these well-mixed and aerated bioreactors, CO

mass transfer did not limit the biomass production
rate. However, it is not known how CO delivery a!ects

the biomass production rate for ¸aminaria saccharina
cell suspensions in the 3 l tubular recycle photo-
bioreactor.
The primary objective of this work is to develop
a batch reactor model for the growth of photosynthetic
cell suspension cultures within the tubular recycle photo-
bioreactor shown in Fig. 1. The model will predict cell
biomass production rate, and nutrient consumption vs.
cultivation time at a given set of process input variables,
including reactor dimensions, tubular section residence
time, intrinsic culture growth parameters, illumination
parameters in the tubular section, and CO mass transfer

parameters in the aeration tank. Carbon dioxide mass
transfer is generally overlooked in the design of tubular
photobioreactor systems. Therefore, the sensitivity
analysis will focus on the CO delivery parameters that

a!ect biomass production, including the mass transfer
coe$cient for CO in the aeration tank, the culture

 G.L. Rorrer, R.K. Mullikin/Chemical Engineering Science 54 (1999) 3153}3162 3155

residence time in the tubular section, and the CO partial that the aeration tank is not illuminated. If < is constant

pressure in the aeration gas. and k is equal to zero, Eq. (1) reduces to
?
dX < f kX
" R R . (4)
dt < #<
2. Model development ? R
Eq. (4) is governed by the initial condition t"t ,

In this section, a batch reactor model for growth of X"X .

photosynthetic cell suspension cultures in a tubular re- Biomass production for photosynthetic suspension
cycle photobioreactor is developed. The growth of cultures requires dissolved carbon dioxide (CO ), essen-

photosynthetic cell suspension cultures requires ex- tial nutrients dissolved in the liquid medium (nitrogen,
ternally supplied sources of both carbon dioxide and phosphorous salts), and light. For example, the overall
light. When the tubular photobioreactor is operated in stoichiometric equation for biomass production by
the total recycle mode, the culture is re-circulated be- photosynthetic ¸aminaria saccharina cultures is (Zhi and
tween the aeration tank, which continuously supplies Rorrer, 1996)
CO , and coiled tubular section, which continuously
 309CO #325H O#16NO\#HPO\
   
supplies light. In this mode of operation, the tubular
recycle photobioreactor shown in Fig. 1 acts as a batch P(CH O) (NH ) (HPO )#341O . (5)
     
reactor with CO mass transfer localized in the aeration
 Since CO is continuously supplied to the culture by the
tank and light transfer localized in the coiled tubular 
aeration gas, the "nal cell density in a batch reactor will
section. Below, the coupled e!ects of nutrient consump- be limited by depletion of an essential nutrient initially
tion, light delivery and transfer, and CO mass transfer
 dissolved in the liquid medium. In photosynthetic sus-
on the biomass growth kinetics are incorporated into the pension cultures, the key limiting nutrient is nitrogen,
model development. supplied to the culture medium in the form of nitrate
(NO\). Under light-saturated growth conditions, the

2.1. Biomass growth kinetics speci"c growth rate for concurrent substrate consump-
tion exhibits saturation kinetics with respect to dissolved
In the batch mode of operation, the accumulation of carbon dioxide concentration (C ) and dissolved nitrate

cell biomass within the entire tubular recycle photo- concentration (C )
,
bioreactor during the exponential phase of growth is
C C
balanced by the rate of biomass production in both the k"k  , :k , (6)
aeration tank and coiled tubular section:  K #C K #C 
  , ,
where K and K are the half-saturation constants for
d(X<)  ,
"k X< #kX< , (1) dissolved CO and nitrate, respectively. For example, the
dt ? ? R 
half-saturation constant for the nitrate consumption (K )
,
in ¸aminaria saccharina intact plants is only 0.0015 mM
where k is the speci"c growth rate of the culture at the (Chapman et al., 1978) vs. 3.0 mM for the initial nitrate
conditions of the tubular section, k  is the speci"c growth
? concentration in the culture medium. Also, the half-satu-
rate of the culture at the conditions of the aeration tank, ration constant for photosynthetic CO "xation in algae
< is the culture volume of the aeration tank, < is the 
? R (K ) is typically less than 0.003 mM vs. 0.015 mM for the

culture volume of tubular section, and X is the cell equilibrium concentration of CO dissolved liquid me-
density at a given time t. In Eq. (1), < is the sum of the 
dium sparged with CO in ambient air (Raven, 1984).
culture volumes in the aeration tank and the tubular 
Consequently, the ¸aminaria saccharina cell suspension
section culture system can approximate zero-order kinetics with
respect to dissolved nutrients, but the speci"c growth rate
<"< #< . (2)
? R is zero when the limiting nutrient is depleted (C "0).
,
For photosynthetic cultures, biomass growth only occurs The "nal cell density achievable in a batch reactor is
if the culture is illuminated. Therefore de"ned by the limiting nutrient initially dissolved in the
liquid medium assuming the cultivation is not limited by
k"k f and k "k f , (3) insu$cient light delivery or cessation of CO delivery. In

R ? ? ? a batch bioreactor of constant liquid volume, the mater-
where f is the fractional photoperiod for tubular ial balance for the limiting nutrient N is given by
R
section illumination system, and f is the fractional
? C "C !(X!X )/> , (7)
photoperiod for the aeration tank illumination system. , ,  6,
Typically, f is equal to 1 so that the tubular section is where > is the stoichiometric biomass yield coe$cient
R 6,
continuously illuminated, whereas f is equal to zero so for nutrient N. From Eq. (5), > is 601.4 mg dry
? 6,
3156 G.L. Rorrer, R.K. Mullikin/Chemical Engineering Science 54 (1999) 3153}3162

light transfer. The attenuation of light through the tubing

wall of thickness l is given by
R
I "I e\IR JR . (10)
R 
The attenuation constant for the silicone tubing (k ) is
R
0.674 cm\ (Mullikin and Rorrer, 1998). Likewise, within
the tubing of inner diameter d, the attenuation of light
through the liquid suspension culture as a function of cell
density (X) is given by
I"I e\IA6B. (11)
R
The attenuation constant for ¸aminaria saccharina cell
suspension culture (k ) is 0.00015 l/mg DCW cm (Mul-
Fig. 2. E!ect of speci"c growth rate (k) on incident light intensity (I )

A
likin and Rorrer, 1998). The mean light intensity (I )
for ¸aminaria saccharina cell suspension culture. Data taken from K
intrinsic bubble-column bioreactor kinetics studies of Rorrer et al. approximates the equivalent light delivery to the culture
(1995) with two-sided illumination (a"2). The data were "tted to in response to light attenuation (Rabe and Benoit, 1962).
Eq. (14) by least-squares nonlinear regression, with k "0.15$0.01 Based on the assumptions above, I for the tubular
 K
(1s, n"4)/day, and I "7.0$3.0 (1s, n"4) lE/m s. As a conservative
I section is approximated by
estimate, I was taken as I #1s"10 lE/m s.
I I


B
I "1/d I(r) dr, (12)
biomass formed p/mmol nitrate consumed for ¸aminaria K

saccharina cell suspension cultures. The "nal cell density where r is the coordinate along the tubing cross section.
(X ) at nutrient depletion (C "0) thus serves as
D , Insertion of Eqs. (10) and (11) into Eq. (12) followed by
a boundary condition for X and is given by integration yields the following expression estimating the
X "X #C > . (8) mean light intensity in the coiled tubular section:
D  , 6,
Photosynthetic cell suspension cultures exhibit satura- I
I (X)"  (e\IR 'R!e\IR JR>IA6B). (13)
tion growth kinetics with respect to light intensity. An K k Xd
A
exponential model of the form
The e!ect of mean light intensity on the speci"c growth
k"k (1!e\''I) (9) rate is therefore approximated by the combination of

Eqs. (9) and (13)
describes the e!ect of light intensity on speci"c growth
rate under nutrient-saturated conditions. For ¸aminaria k"k (1!e\'K 6'I). (14)

saccharina cell suspension cultures, k is equal to

0.15 day and I is equal to 10 lE/m s (see Fig. 2), using Further combination of Eqs. (4) and (14) leads to a di!er-
I
data obtained from previous intrinsic growth kinetics ential model which describes the e!ect of light delivery
studies in an illuminated bubble column bioreactor (Ror- and transfer on the biomass growth kinetics
rer et al., 1995).
dX < f kX < f k (1!e\'K 6'I) X
" R R " R R  , (15)
2.2. Ewect of light transfer on biomass growth kinetics dt < #< < #<
? R ? R
where I (X) is de"ned by Eq. (13). Eq. (15) is valid under
Light transfer to the photosynthetic culture occurs K
conditions where the biomass growth kinetics are not
only within the tubular section. The coiled tubular sec- limited by CO mass transfer. The e!ects of CO mass
tion is uniformly illuminated by a lamp, which is verti-  
transfer on the biomass growth kinetics are described
cally aligned along the center axis of the coil. In the next.
tubular section, the coiled tubing is stacked, the curva-
ture of the tubing is smaller relative to the distance from 2.3. Ewect of CO delivery on biomass growth kinetics
the light source to the tubing wall, and the radius of the 
coiled tubing section is much larger than the radius of the Dissolved carbon dioxide (CO ) is the carbon source
tubing itself. Consequently, a planar approximation for 
for biomass production in photosynthetic cell suspension
light transfer through the coiled tubular section is rea- cultures. The volumetric consumption rate of dissolved
sonable. The transfer of light through the tube is CO by the photosynthetic cell suspension culture ap-
attenuated by the tubing wall and by the suspension 
proximates zero-order kinetics of the form
culture inside the tubing, acting as two resistances in
series. The reduction in light intensity is described by the kX
Q " . (16)
Beer}Lambert law relationship along the path length for  >
6!-
 G.L. Rorrer, R.K. Mullikin/Chemical Engineering Science 54 (1999) 3153}3162 3157

From Eq. (5), the biomass yield coe$cient for CO Based on Eq. (23), X increases as C*, k a, and v in-
 A  * 
(> ) for ¸aminaria saccharina cultures is 31 mg of dry crease.
6!-
biomass formed/mmol CO consumed. The delivery of As the cell density (X) in the bioreactor increases with

CO to the tubular recycle photobioreactor occurs only time, the volumetric CO consumption rate increases by
 
in the aeration tank. Interphase mass transfer of CO Eq. (16). When X*X , the CO mass transfer rate will
 A 
from the aeration gas to the culture liquid medium sets not meet the required CO consumption rate, and the

the CO delivery rate to the aeration tank, and the liquid biomass growth kinetics will be limited by the CO mass
 
"lm resistance is controlling since CO is sparingly sol- transfer rate. The biomass growth kinetics are now de-

uble in water. Although the cell density (X) increases with scribed by
time in batch culture, CO consumption and mass trans-
 dX < f kX k aC*< > f
fer rates are at a nominal steady state for a given value of " R R " *  ? 6!- R (24)
dt < #< (< #< ) (k a< /v #1)
X. The steady-state material balance for CO in the
 ? R ? R * ? 
liquid phase of the well-mixed aeration tank of constant which is obtained by rearrangement and insertion of
liquid volume is Eq. (22) into Eq. (4).
kX The uptake of CO from the aeration gas to the liquid
v C #k a(C*!C )< !v C ! ? < "0. (17) 
 * *  - ?  - > ? culture is enhanced if the medium is alkaline. Photosyn-
6!-
thetic marine plant cell suspension cultures grow best in
Since the aeration tank is not illuminated, k is equal to
? arti"cial seawater meidum at pH 8}9, where CO speci-
zero. At this condition, the outlet concentration of dis- 
ates to bicarbonate (HCO\) and carbonate (CO\)
solved CO in the aeration tank (C ) is  
 -
CO (g)  CO (aq),
k a< C*#v C  
C " * ?   * . (18)
- k a< #v CO (aq)#H O  HCO\#H>,
* ?    
The dissolved CO concentration in the liquid medium in HCO\  CO\#H> (25)
  
equilibrium with the partial pressure of CO (P ) in the
  or
aeration gas is given by Henry's law:
CO (aq)#OH\  HCO\ .
C*"P /H . (19)  
  
The pK values for HCO\and CO\ dissociation are
Since the tubular section is not aerated, dissolved CO is
 ?  
consumed by the culture as it #ows down the length of 6.05 and 9.23, respectively in seawater of 35 ppt salinity
at 153C, and H for CO is 0.023 atm mmol\ l at the
the tube. At a given cell density (X), the material balance  
for dissolved CO on a di!erential volume element of the same conditions (Raven, 1984). From these mass-action
 expressions, if ambient air containing 350 ppm CO is
continuously illuminated tubular section is 
bubbled into seawater medium at pH 8.0 and 153C, then
dC kX nd the equilibrium dissolved CO , HCO\ and CO\ con-
"! (20)   
dz > 4v centrations are 0.015, 1.2, and 0.071 mM, respectively,
6!- 
whereas at pH 7.0 equilibrium concentrations of HCO\
which upon integration from z"0 to z"¸ over the 
and CO\ are only 0.12 and 0.00071 mM, respectively.
length of the tube results in 
At high pH, this bicarbonate reservoir can serve as a bal-
kX nd kX < last for dissolved CO and potentially help avoid CO
 
C "C ! ¸"C ! R. (21)
* - > 4v - > v limitation during peak CO demand. However, CO
 
6!-  6!-  speciation was not accounted for in the model develop-
If C goes to zero, then the process is limited by an ment.
*
insu$cient CO delivery rate. At C equal to zero,
 *
combination of equations (18) and (21) to eliminate 2.4. Specixc biomass production rate
C yields
-
k aC*> < /< For either rate-limited growth (Eq. (15)) or mass trans-
kX" *  6!- ? R . (22) fer limited growth (Eq. (24)), the speci"c biomass produc-
k a< /v #1
* ?  tion rate (r ) is de"ned as
Eq. (22) describes the inter-relationships between all pro- 6
r "1/X dX/dt. (26)
cess variables at the point of CO mass transfer rate
 6
limitation. For example, from Eq. (22), the cell density at
which biomass growth becomes CO mass transfer rate 2.5. Solution of model equations

limited (X ) is
A Prediction of the cell density as a function of time
k aC*> < and process input variables for the tubular recycle
X" *  6!- ? . (23)
A <k (k a< /v #1) (1!e\'K 6A'I) photobioreactor in the batch mode of operation is
R  * ? 
3158 G.L. Rorrer, R.K. Mullikin/Chemical Engineering Science 54 (1999) 3153}3162

Table 1
Summary of process model parameters

Tubular reactor Tubular section illumination

d 0.794 cm k 0.00015 l/mg DCW cm

A
¸ 0.159 cm k 0.674 cm\
R R
¸ 40.4 m I 30.0 lE/m s

< 2.0 l I 27.0 lE/m s
R R
v 0.5 l/min f 1.0 h on/24 h photoperiod
 R
Aeration tank Biomass growth
P 0.00035 atm k 0.15 1/day
 
H (153C) 0.023 atm l/mmol I 10 lE/m s
 I
C* 0.015 mmol CO /l > 31.1 mg DCW/mmol CO
  6!- 
< 1.0 l > ‚ 601.4 mg DCW/mmol NO\
? 6, 
Initial conditions Final conditions
X 100 mg DCW/l X 1904 mg DCW/l
 D
C 3.0 mmol/l C 0 mmol/l
, ,D

accomplished by numerical integration of the biomass
growth rate equations using the fourth-order Runge}
Kutta method using a step size of 0.2 day. Process model
inputs shown in Table 1 include tubular reactor para-
meters, tubular section illumination parameters, aeration
tank parameters, biomass growth parameters, and initial
conditions for cell density and nutrient concentration.
The representative biomass growth parameters given in
Table 1 are for photosynthetic cell suspension cultures of
the macrophytic brown alga ¸aminaria saccharina (Zhi
and Rorrer, 1996). All other parameters given in Table
1 are taken from the design of a 3 l tubular recycle
photobioreactor recently described by Mullikin and Ror-
rer (1998). Prior to integration, X at the desired set of
A D
process input variables is determined by "nding the root
of Eq. (23) using Newton's method. If X (X (X ,
 A D 6
then Eq. (15) is integrated until X"X . At X"X ,
A A
biomass growth kinetics are limited by CO mass trans-

fer, and Eq. (24) is integrated until X"X . Values for
D
C (Eq. (7)), and r (Eq. (26)) are also computed. At X"
, 6
X , nutrient limitation in the liquid medium occurs and
D
biomass growth ceases. If X )X , then the biomass
A 
growth kinetics will always be limited by the CO mass

transfer rate. If very high cell densities are achieved, then
by Eq. (14) the speci"c growth rate (k) may decrease
signi"cantly in response to light attenuation, particularly
if I (X) becomes signi"cantly lower than I , and the
K I
biomass growth kinetics will once again be subject to
Eq. (15).
3. Results and discussion
3.1. Base case model predictions
The "nal model for the tubular recycle photobioreac-
tor predicts cell density (X), limiting dissolved nutrient
concentration (C ), mean light intensity in the tubular
,
section (I ), and speci"c biomass production rate (r ) as
K 6
a function of cultivation time for batch cultivation of
a photosynthetic cell suspension culture in the total re-
cycle mode of operation. Predictions for X, C , I and
, K
r vs. cultivation time for the base-case model input
6
parameters given in Table 1 are presented in Figs. 3a and
b. At these conditions, the transition from rate-limited to
CO delivery rate limited growth occurs at X "757 mg
 A
DCW/l. In the tubular recycle photobioreactor, the at-
tenuation of light during the cultivation process is mod-
est, with I decreasing from 26.8 lE/m s at inoculation
K
(X "100 mg DCW/l) to 24.1 lE/m s at a "nal cell

density (X "1904 mg DCW/l). Both of these values are
at least two times greater than I . As shown in Fig. 3b,
I
the speci"c biomass production rate (r ) for rate-limited
growth within the regime X (X(X is not signi"-
 A
cantly a!ected by light attenuation. Therefore, growth of
the photosynthetic suspension culture in the tubular
photobioreactor is operating at saturation kinetics with
respect to illumination and is not limited by light trans-
fer. This result is attributed to the small light path
(d"0.794 cm) of the coiled tubular section. In fact, the
tubing wall attenuates light about equally to the culture
itself, since I is 30 lE/m s incident to the tube wall

whereas I is 27 lE/m s incident to the culture inside
R
the tubing. A sensitivity analysis on Eq. (13) further
reveals that I does not approach I until d"4 cm and
K I
X"4000 mg DCW/l for the input parameters given in
Table 1.
Based on the above discussion, the tubular recycle
photobioreactor e!ectively delivers light to the photo-
synthetic cell suspension. However, biomass production
rate in the tubular recycle photobioreactor is often lim-
ited by the delivery of CO to the photosynthetic liquid

suspension culture. The e!ects of CO delivery on

biomass production are described next.
 G.L. Rorrer, R.K. Mullikin/Chemical Engineering Science 54 (1999) 3153}3162
Fig. 3. Predicted cell density (X), mean light intensity (I ), limiting
K not possible at this asymptote.
nutrient concentration (C ), and speci"c biomass production rate (r ) in
, 6
tubular recycle photobioreactor for the process input parameters given
in Table 1. (a) X and I vs. cultivation time; (b) C and r vs. cultivation
K , 6
time.
3.2. Ewect of CO2 delivery on biomass production
Four process parameters have a signi"cant impact on
CO delivery to the tubular recycle photobioreactor: the

interphase CO mass transfer coe$cient in the aeration

tank (k a), the residence time of the liquid suspension
*
culture in the tubular section (</v ), the ratio of the
R 
tubular section culture volume to the aeration tank cul-
ture volume (</< ), and the CO partial pressure in the
R ? 
aeration gas (P ), which sets C* . Below, the sensitivity of
 
k a, < /v , < /< and C* to the cell density vs. time pro"le
* R  R ? 
in batch operation of the tubular recycle photobioreactor
is described for an interphase mass-transfer-based pro-
cess model.
The e!ect of the volumetric mass transfer coe$cient
for CO in the aeration tank (k a) on the cell density vs.
 *
time pro"le is presented in Fig. 4 for the input parameters
given in Table 1. At a k a value of 2.5 h\, the cultivation
*
is CO mass transfer rate limited at inoculation
 A
(X 'X ), and so the cell density vs. time pro"le is linear
A 
as predicted by Eq. (24). For comparison, the cell density
vs. time pro"le for the rate-limited cultivation (X 'X )
A D
3159
Fig. 4. E!ect of volumetric mass transfer coe$cient for CO in aer-

ation tank (k a) on the predicted cell density (X) vs. cultivation time
*
curve in the tubular recycle photobioreactor for the process input
variables given in Table 1.
governed by Eq. (15) is also provided in Fig. 4. As k a
*
increases, X increases by Eq. (23) and biomass produc-
A
tion is rate limited until X"X . The biomass produc-
A
tion rate increases as k a increases from 2.5 to 100 h\,
*
but past a k a value of 100 h\ the enhancement for the
*
biomass production rate is minimal. In Eq. (23), as
k aPR, X is 1219 mg DCW/l for the input parameters
* A
given in Table 1, which is less than X value of
D
1904 mg DCW/l. Therefore, rate limited growth is still
The e!ect of tubular section residence time (< /v ) on
R 
the cell density vs. time pro"le is presented in Fig. 5 for
the input parameters given in Table 1. For comparison,
the cell density vs. time pro"le for the rate-limited culti-
vation (X 'X ) governed by Eq. (15) is also provided in
A D
Fig. 5. As the residence time of the culture within the
tubular section of photobioreactor decreases from 10 to
2 min, the biomass production rate increases and asymp-
totically approaches the rate-limited cultivation case. To
adjust the residence time, the culture #owrate was in-
creased from 100 to 2000 cm/min, which corresponds to
Reynolds number ranging from 224 to 4474 assuming the
culture approximates the properties of water at 153C.
Decreasing the culture residence time inside the tubular
section by increasing the culture #owrate v decreases the

single-pass conversion of CO and thus helps to avoid

complete consumption of CO by the photosynthetically

active culture within the non-aerated tubular section.
However, under CO delivery limited growth, increasing

v increases the overall biomass production rate since the
M
cumulative CO consumption rate over many passes in

total recycle operation increases. As v increases, X in-
 A
creases by Eq. (23) and biomass production is rate limited
until X"X . In Eq, (23) as v PR, X is 758 mg DCW/l
 A
for the input parameters given in Table 1, which is less
than X of 1904 mg DCW/l. Therefore, rate-limited
D
culture growth is still not possible at this asymptote.
3160 G.L. Rorrer, R.K. Mullikin/Chemical Engineering Science 54 (1999) 3153}3162

Fig. 5. E!ect of tubular section culture residence time (< /v ) on the
R 
predicted cell density (X) vs. cultivation time curve in the tubular
recycle photobioreactor for the process input variables given in Table 1.
Fig. 6. E!ect of the ratio of tubular section to aeration tank culture
volume (< /< ) on the predicted cell density (X) vs. cultivation time
R ?
curve in the tubular recycle photobioreactor for the process input
variables given in Table 1.

The e!ect of the ratio of tubular section culture volume

to aeration tank culture volume (< /< ) has a complex
R ?
e!ect on the cell density vs. time pro"le, as shown in
Fig. 6. Based upon the input parameters given in Table 1,
the overall biomass production rate is optimal at < /<
R ?
equal to 2. An optimum value for < /< exists because
R ?
< /< has opposing a!ects on rate-limited culture growth
R ?
through Eq. (15) and CO delivery limited culture growth

through Eq. (24). According to Eq. (23), X decreases as
A
< /< increases. Therefore, rate-limited growth is favored
R ?
at low < /< ratios, whereas CO delivery limited growth
R ? 
occurs at high < /< ratios. At < /< equal to 0.5, the
R ? R ?
biomass production is rate-limited. However, at < /<
R ?
equal to 10, culture growth is CO delivery limited short-

ly after inoculation (X "215 mg DCW/l). Nevertheless,
A Fig. 7. E!ect of aeration gas partial pressure (P ) relative to the ambi-
as shown in Fig. 6 the CO -limited process at < /< equal 
 R ? ent CO partial pressure of CO in air (35 Pa) on the predicted cell
 
to 10 has a higher biomass production rate than the density vs. cultivation time curve in the tubular recycle photobioreactor
rate-limited process at < /< equal to 0.5.
R ? for the process input variables given in Table 1.
The e!ect of CO partial pressure in the aeration gas

(P ) on the cell density vs. time pro"le is presented in

Fig. 7 for the input parameters given in Table 1. The tank. Model simulations show that at the input para-
ambient CO partial pressure in air is 35 Pa. Increasing meters given in Table 1, the tubular recycle photo-

P increases C* by Eq. (19) and hence increases X by bioreactor is operating at saturation growth kinetics with
  A
Eq. (23). The biomass production rate also increases with respect to light. The model predicts a critical cell density
increasing P . However, rate-limited growth occurs at at which photosynthetic biomass production switches

two times ambient CO partial pressure, and no further from a rate-limited process to a CO delivery limited
 
improvement in biomass production rate is observed at process. Rate-limited growth proceeds only to this criti-
higher CO partial pressures. cal cell density, and then further growth is CO mass
 
transfer limited until the "nal cell density at nutrient
limitation is achieved. Model simulations reveal that
4. Concluding remarks CO mass transfer coe$cient in the aeration tank (k a),
 *
the residence time of the culture in the tubular section
A batch reactor model was developed to predict bio- (< /v ), the ratio of tubular section to aeration tank liquid
R 
mass growth kinetics for cultivation of photosynthetic volume (< /< ), and the partial pressure of CO in the
R ? 
cell suspension cultures in the tubular recycle photo- aeration gas (P ) a!ect the overall biomass production

bioreactor shown in Fig. 1. This model uniquely couples rate by controlling the CO delivery mode to the tubular

culture illumination parameters in the tubular section to photobioreactor. Furthermore, sensitivity analysis re-
interphase CO mass transfer parameters in the aeration veals that CO delivery can be optimized for a particular
 
 G.L. Rorrer, R.K. Mullikin/Chemical Engineering Science 54 (1999) 3153}3162
cell culture to maximize biomass production rate and

target cell density, as each variable mentioned above
possesses an optimum or an asymptote. Future model
improvements may include the use of saturation growth
kinetics for dissolved CO and limiting nutrient for more
 R
a accurate estimation of k as opposed to the reasonable
simpli"cation of zero-order growth kinetics, and consid-
eration of enhanced mass transfer of CO at high pH

cultivations.
Acknowledgements
This work was supported by grant no. NA36RG0451
(project no. R/BT-8) from the National Oceanic
and Atmospheric Administration to the Oregon State
University Sea Grant College Program under the
National Sea Grant Program Marine Biotechnology
Initiative.
Notation
C concentration of dissolved CO in tubular
 
section at axial position z, mmol/l
C* concentration of dissolved CO in equilib-
  
rium with the CO partial pressure in the

aeration gas, mmol/l
C concentration of dissolved CO exiting
* 
tubular section or entering aeration tank,
mmol/l
C concentration of dissolved CO exiting aer-
- 
ation tank or entering tubular section,
mmol/l
C concentration of dissolved nutrient in liquid
,
medium, mmol/l
C initial concentration of dissolved nutrient in
,
liquid medium, mmol/l
d inner diameter of tubing, cm
f fractional photoperiod for aeration tank illu-
?
mination system, h light on/24 h
f fractional photoperiod for tubular section il-
R
lumination system, h light on/24 h
H Henry's law constant for dissolved CO in
  ?
arti"cial seawater medium, atm l/mmol
k light attenuation constant for liquid cell sus-
A
pension culture, l/mg DCW cm

k a volumetric mass transfer coe$cient for
*
CO interphase mass transfer in aeration

tank, 1/h
k light attenuation constant for translucent sili-
R
cone tubing, 1/cm
K half-saturation constant for dissolved CO ,
 
mmol/l
K half-saturation constant for limiting nutrient,
,
mmol/l
3161
I light intensity corresponding to 0.673 k ,
I
lE/m s
I mean light intensity delivered to liquid sus-
K
pension culture, lE/m s
I light intensity (irradiance) incident to liquid
suspension culture inside tubing, lE/m s
I light intensity (irradiance) incident to tubing

wall in coiled tubular section, lE/m s
¸ length of coiled tubing in tubular section, m
l thickness of silicone tubing, cm
R
P partial pressure of CO in the aeration gas,
 
Pa or atm
Q volumetric consumption rate of dissolved
CO by cell suspension culture, mmol/l h

r speci"c biomass production rate (1/day)
6
t cultivation time, days
v volumetric #owrate of liquid suspension

culture through coiled tubular section,
l/min
< total liquid volume of tubular recycle photo-
bioreactor, l
< liquid volume of aeration tank, l
?
< liquid volume of coiled tubular section, l
R
X cell density, mg DCW/l
X cell density in tubular photobioreactor where
A
biomass growth kinetics become CO trans-
fer rate limited at a given set of process condi-
tions, mg DCW/l
X "nal cell density at nutrient depletion,
D
mg DCW/l
X initial cell density, mg DCW/l

> biomass yield coe$cient based on CO con-
6!- 
sumption, mg DCW/mmol CO

> biomass yield coe$cient based on limiting
6,
nutrient consumption, mg DCW/mmol N
z axial position down length of tubing, m
Greek letters
a number of illumination planes
k speci"c growth rate of liquid suspension
culture at conditions in tubular section,
1/day
k speci"c growth rate of liquid suspension
culture at conditions in aeration tank,
1/day
k speci"c growth rate of liquid suspension cul-
ture at saturation light intensity, 1/day
Abbreviations
DCW dry cell weight
mM mmol/l
lE micro Einstein, equivalent to 1.0 lmol of
photon quanta in photosynthetically active
irradiance (400}700 nm wavelength)
3162 G.L. Rorrer, R.K. Mullikin/Chemical Engineering Science 54 (1999) 3153}3162
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