Bioresource Technology 136 (2013) 689–696

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Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Modeling microalgal growth in an airlift-driven raceway reactor

Balachandran Ketheesan, Nagamany Nirmalakhandan ⇑
Civil Engineering Department, New Mexico State University, Las Cruces, NM 88002, USA

h i g h l i g h t s g r a p h i c a l a b s t r a c t

" Mathematical model for algal

biomass growth in an airlift-driven
raceway.
" Validated model with growth data
on Nannochloropsis and Scenedesmus.
" Validated model with growth data
from indoor and outdoor conditions.
" Predicted biomass densities agreed
with measured ones with r2 > 0.96.

a r t i c l e i n f o a b s t r a c t

Article history: In previous proof-of-concept studies, feasibility of a new airlift-raceway configuration and its energetic
Received 20 November 2012 advantage and improved CO2 utilization efficiency over the traditional raceways and photobioreactors
Received in revised form 12 February 2013 have been documented. In the current study, a mathematical model for predicting biomass growth in
Accepted 14 February 2013
the airlift-raceway reactor is presented, which includes supply and transfer of CO2 and the synergetic
Available online 28 February 2013
effects of light, CO2, nitrogen, and temperature. The model was calibrated and validated with data from
prototype scale versions of the reactor on two test species: Nannochloropsis salina and Scenedesmus sp.,
Keywords:
cultivated under indoor and outdoor conditions. Predictions of biomass concentrations by the proposed
Algal growth model
Airlift-raceway reactor
model agreed well with the temporal trend of the experimental data, with r2 ranging from 0.96 to 0.98,
Carbon dioxide transfer p < 0.001. A sensitivity analysis of the 10 model parameters used in this study revealed that only three of
Simulation them were significant, with sensitivity coefficients ranging from 0.08 to 0.13.
Ó 2013 Elsevier Ltd. All rights reserved.

1. Introduction
Microalgae has been reported to be advantageous over conven-
tional feedstocks such as oil crops for biodiesel production due to
its higher solar energy yield, higher biomass yield, competitive
net energy gains, and its potential to utilize waste streams such
as carbon dioxide from flue gases and nutrients from wastewaters
(Weyer et al., 2010). However, recent techno-economic evaluations
of biodiesel production based on the traditional algal cultivation
systems, such as the open raceways, indicate that they may not
be energy-efficient and cost-effective for biodiesel production
⇑ Corresponding author. Tel.: +1 575 646 5378; fax: +1 575 646 6049.
E-mail address: nkhandan@nmsu.edu (N. Nirmalakhandan).
(Singh et al., 2012). This has fueled the development of engineered
photobioreactors and hybrid cultivation systems capable of higher
volumetric biomass productivity per unit energy input.
An airlift-raceway configuration has been proposed as a possi-
ble improvement to the traditional paddlewheel-driven raceway
design for improved energy-efficiency and CO2 utilization effi-
ciency. In previous proof-of-concept studies (Ketheesan and Nir-
malakhandan, 2011, 2012), it was shown that the energy
required for maintaining typical circulation velocities of 8–
14 cm s1 in paddlewheel-driven raceways could be reduced 40–
80% in this airlift-raceway configuration. Volumetric biomass pro-
ductivities (0.085 dry g L1 day1) and CO2 utilization efficiencies
(33%) achieved in this configuration were comparable to or better
than those reported for paddlewheel-driven raceways. Biomass

0960-8524/$ - see front matter Ó 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.biortech.2013.02.028
690 B. Ketheesan, N. Nirmalakhandan / Bioresource Technology 136 (2013) 689–696
productivity per unit energy input in this configuration was shown
to be higher than those reported for engineered photobioreactors
(0.60–0.69 dry g W1 day1 vs. 0.10–0.51 dry g W1 day1) (Kethe-
esan and Nirmalakhandan, 2011, 2012).
Based on the above findings, it is hypothesized that with further
optimization in terms of energy and material inputs and operating
conditions, the performance of the airlift-raceway design could be
enhanced further. Since process models capable of describing bio-
mass growth in terms of the process parameters could be benefi-
cial in predicting process performance, optimizing operating
conditions, identifying sensitive parameters, and in scaling up
reactor systems (Geider et al., 1998), this study was undertaken
to develop, calibrate, and validate a mathematical model for the
airlift-raceway reactor.
While the growth of outdoor cultures could be influenced by a
number of parameters independently or synergistically, most out-
door raceway systems have been designed and operated to attain
the upper limit of photosynthetic yields, with excess supply of
CO2 and nutrients so that growth is dependent only on two uncon-
trollable environmental variables i.e. sun light (intensity, duration)
and temperature (Goldman, 1979). Following this line of thought,
several modeling studies have been reported in the literature with
the intent to establish the irradiance profile inside the bulk culture
with respect to the bioreactor configuration and to predict biomass
productivity by photosynthetic mechanisms as a function of the
incident irradiance and temperature (Sukenik et al., 1987, 1991;
Molina Grima et al., 1994). For instance, a theoretical framework
has been established by Goldman (1979) to predict the photosyn-
thetic yield of microalgae as a function of sunlight intensity inci-
dent on the open raceway. Sukenik et al. (1987) have modeled
light attenuation effects on algal productivity in a raceway and
simulated daily biomass production, and refined it to a determin-
istic model (Sukenik et al., 1991) including photoadaptation, gross
photosynthesis, and respiration as a function of irradiation and
temperature. Besides, several theoretical approaches (Weyer
et al., 2010) have also been proposed to estimate the upper limit
of algal production in response to solar energy input.
While existing models provide a basis for optimizing photosyn-
thetic yields, such models may not be applied when growth is lim-
ited by CO2 or any other major nutrient. Since carbon content of
biomass is about 50% (Becker, 1994), supply of carbon to the algal
cultures could possibly limit algal growth due to poor transfer of
CO2 from ambient air via surface aeration; or low transfer effi-
ciency of CO2-sparged systems. The traditional approach of excess
supply of CO2 may not be a sustainable option since delivery of CO2
represents a major component of cultivation costs (Becker, 1994).
In this context, the need for optimizing the mass input of CO2
either under light limiting or non-light limiting condition has made
modeling the supply and transfer of CO2 in the airlift-raceway
reactor necessary.
In this work, the use of an airlift column in an open raceway for
the supply and transfer of CO2 is modeled, integrating it with a
model for biomass growth incorporating the synergetic effects of
growth-limiting parameters such as light, CO2, nitrogen and tem-
perature. In contrast to existing models in the literature for open
raceways, the current study will provide a useful benchmark to reg-
ulate the input of CO2 in response to temporal operational variables
and culture conditions. The application of this proposed model
relating the CO2 supply is that, on one hand, CO2 supply can be reg-
ulated on the basis of temporal light fluctuations even when the
photosynthesis is solely limited by light; and, on the other hand,
maximum limit of biomass yield can be attained via augmenting
the supply of photosynthetic carbon towards biomass synthesis
when CO2 is the primary photosynthetic-yield determinant.
In order to demonstrate the CO2-dependence of microalgal
growth and to rationalize the model expressions, selective culture
conditions were maintained at different CO2-to-air ratios. The pro-
posed model was calibrated and validated using growth data of
two algal species Nannochloropsis salina (N. salina), a marine algae,
and Scenedesmus sp., a fresh water algae, cultivated in two proto-
type versions of the airlift reactor, under indoor and outdoor
conditions.
2. Methods
2.1. The airlift-raceway reactor
As shown in Fig. 1, the proposed airlift-raceway system consists
of an open raceway integrated with an airlift configuration which
enables liquid circulation and CO2 transfer. Ambient air is sparged
from the bottom of the riser of the airlift section, generating a
hydraulic head to maintain liquid circulation in the raceway. Sup-
plemental CO2 is bubbled at the mid-depth of the downcomer so
that the downward flow of the broth through the downcomer re-
tains the rising CO2 bubbles over a prolonged period and carries
them towards the riser side, improving the bubble residence time
and hence the transfer of CO2 into the broth. Details of the reactor
design have been reported elsewhere (Ketheesan and Nirmalakh-
andan, 2012). Experimental data for calibrating and validating
the model were collected from two such reactors, constructed with
different riser heights: Reactor 1, with a riser height of 48 cm and
culture volume of 20 L; and Reactor 2, with a riser height of 72 cm
and culture volume of 23 L.
2.2. Microorganisms and culture media
Growth data on a marine microalgae N. salina (Eustigmatophy-
ceae) and a fresh water microalgae Scenedesmus sp. (Chlorophy-
ceae) were collected to calibrate and validate the model. N. salina
was cultivated in modified f/2 (Arudchelvam and Nirmalakhandan,
2012) saline media with nitrate level of 1 mM NaNO3 in the labo-
ratory studies and with 5 mM NaNO3 in the outdoor studies; Scene-
desmus sp. was cultivated in fresh water Bold’s basal medium
(Bischoff and Bold, 1963) with nitrate level of 3 mM NaNO3.
2.3. Reactor operation
Tracer pulse injection method (Verlaan et al., 1989) was de-
ployed to determine the liquid circulation velocity and assess mix-
ing characteristics in the airlift-raceway reactor (see
Supplementary materials).
Biomass growth data were collected in indoor and outdoor tests
conducted in batch mode, under sparging with ambient air
Fig. 1. Schematic of the airlift-raceway reactor.
 B. Ketheesan, N. Nirmalakhandan / Bioresource Technology 136 (2013) 689–696 691

continuously, at a rate of 2.4 L min1. Temperature and pH were via riser/downcomer, a bubble plume was formed immediately
measured continuously (Mettler Toledo M300) and OD 750 was above the spargers, although the bulk liquid phase in the main
measured daily. Light levels were measured using a Quantum raceway was completely mixed (McGinnis and Little, 2002).
PAR meter (Apogee MQ-200). Correlations between OD 750 and The number of gas bubbles per unit area per unit time No (m2 -
dry density of biomass (g L1) were used to estimate the dry den- s1) formed at the sparger can be given as
sity of algal cultures (Ketheesan and Nirmalakhandan, 2012).
Qg
Indoor studies were conducted under constant artificial illumi- No ¼ ð2Þ
V b Að1  eÞ
nation provided continuously with two 40 W fluorescent lights
(80–90 lE m2 s1). CO2 was supplied continuously via CO2 mass where Qg is the volume flow rate of gas at the sparger (m3 s1), A is
flow controllers (Mass-Trak model 810 C) at different rates. the area of cross section of the riser or downcomer (m2), Vb is the
Growth data were collected under sparging with ambient air and volume of one gas bubble (m3) and e is the gas hold-up.
with supplemental CO2 injection, under the following CO2-to-air The total number of bubble flux per unit plume volume, N, can
ratios: N. salina with 0.25%, 0.5%, 1%, and 2% (vol vol1) in Reactor therefore be expressed as:
1; Scenedesmus sp. with 0.25%, 0.5%, 1%, and 3% (vol vol1) in Reac-
No
tor 2. N¼ ð3Þ
Outdoor tests were conducted only with N. salina, in a green- ðw þ wb Þ
house under natural irradiance where, the average incident PAR where w is the liquid velocity (m s1) and wb is the gas terminal
on the free surface of the raceway was recorded every 30 min. velocity (m s1).
Temperature inside the greenhouse was controlled at 15 °C during The total surface area of the bubbles per unit plume volume, a
night and at 22 °C during day light period. Reactor 1 was run with- (m2 m3) can be expressed as
out any supplemental CO2 sparging; while Reactor 2 was run under
pH-controlled supplemental CO2 sparging, with the pH set at 7.5. N
a ¼ 4pr 2 N ¼ 4pr 2 ð4Þ
In the pH-controlled tests, cumulative mass flow rate of CO2 was ðw þ wb Þ
recorded by a mass flow meter (Aalborg), and the partial pressure where r is the radius of gas bubbles formed at the sparger (m).
of CO2 in the liquid phase was continuously recorded by a dis- Incorporating mass transfer coefficient of CO2, KLCO2 (m s1) at
solved CO2 meter (Mettler Toledo M700). liquid side, an expression for overall mass transfer coefficient,
KLaCO2 (s1) in the riser/downcomer side can be derived as
3. Model development
N
K L aCO2 ¼ 4pr2 K LCO2 ð5Þ
ðw þ wb Þ
3.1. Modeling CO2 transfer
Here, mass transfer coefficient of O2 at liquid side was adopted
Dissolved CO2 in the liquid phase is modeled first by setting up from Wuest et al. (1992) to estimate the mass transfer coefficient
mass balance equations considering the following processes, of CO2, KLCO2 (m s1) using the following equation (Tamimi et al.,
assuming the bulk liquid phase to be well-mixed: (i) rate of trans- 1994).
fer of CO2 from the atmosphere across the free surface, Ma
K LCO2 ¼ WCO2 K LO2 ð6Þ
(mol s1); (ii) rate of transfer of CO2 from the sparging air in the ri-
ser, Mr (mol s1); (iii) rate of transfer of CO2 from the supplemental and
CO2 supply in the downcomer, Ms (mol s1); and (iv) rate of con-  0:5
sumption of CO2 by microalgae, Mc (mol s1). DCO2
Wco2 ¼ ð7Þ
DO2
3.1.1. Modeling rate of transfer of CO2 from the atmosphere across the where DCO2 is the diffusivity of CO2 in water (cm2 day1) and DO2 is
free surface (Ma) the diffusivity of O2 in water (cm2 day1).
The transfer of CO2 from the atmosphere across the free surface,
Ma, (mol s1) can be expressed as follows. 3.1.2.2. Modeling rate of transfer of CO2 from the sparging air in the
M a ¼ K La;air ðC L;air  C 1 ÞV t ð1Þ riser (Mr). The rate of transfer of CO2 via the riser is modeled con-
sidering an elemental mass balance and integration, with the fol-
where KLa,air is the surface mass transfer coefficient (s1), lowing assumptions: the gas phase flows in a plug flow manner
ðC La;air  C 1 Þ is the CO2 concentration driving force in liquid phase, while the liquid phase is completely mixed; and the pressure of
mol m3 and Vt is the total reactor volume, m3. The mass transfer the gas phase and the molar gas flow rate remain constant with
coefficient of CO2 through the surface of the raceway channel, KLa,air depth. Thus, the elemental mass balance on CO2 in the gas phase
was taken as 1.24E4 s1 (Weissman et al., 1988). in the riser can be expressed as
 
d pðeAdzÞ
3.1.2. Modeling rate of transfer of CO2 in riser and downcomer y ¼ M in  Mout  dM r ð8Þ
dt RT
3.1.2.1. Modeling mass transfer coefficient in riser/downcomer. Since
the proposed airlift-raceway reactor involves multiple modes for where p is total pressure in the gas phase (atm), e is the gas holdup
transferring CO2 from gaseous phase to liquid phase, the discrete in the riser (), A is the area of riser (m2), y is the mole fraction of
bubble plume model reported by Wuest et al. (1992) and McGinnis CO2 in the gas phase (), R is the Ideal Gas constant (atm m3 mol1 -
and Little (2002) for modeling oxygen transfer in diffused-bubble K1), T is the absolute temperature (K), Min is the molar rate of in-
system are adapted to model CO2 mass transfer coefficient, KLaCO2 flow of CO2 into the element (mol s1), Mout is the molar rate of
(s1) in riser/downcomer. outflow of CO2 from the element (mol s1), dMr is the molar rate
The gas phase was assumed to flow in a plug flow manner of transfer of CO2 from the gas phase to the liquid phase within
through the riser and the downcomer. The initial bubble size distri- the element (mol s1).
bution and the rate of bubble formation were assumed to be con- The rate of transfer of CO2 from the gas phase to the
stant and the distribution of the bubble sizes, represented by the liquid phase, dMr, can be modeled following the two-film theory
Sauter-mean diameter. It was assumed that when bubbles sparged as:
692 B. Ketheesan, N. Nirmalakhandan / Bioresource Technology 136 (2013) 689–696

dM r ¼ K L aCO2 ð1  eÞAdz½C 1  C 1  ð9Þ dX

¼ lX  k d X ð18Þ
1 ⁄ dt
where KLaCO2 is the mass transfer coefficient of CO2 in riser (s ), C1
is the liquid phase concentration of CO2 that is in equilibrium with where kd is the biomass decaying rate and, l is the specific growth
the gas phase (mol m3), and C1 is the liquid phase concentration of rate that can be expressed by a Michaelis–Menten type relationship
CO2 (mol m3). modified to include limiting effects of carbon dioxide, light, temper-
Combining Eqs. (8) and (9), ature and nitrogen, as follows:
   " #" #
d pðeAdzÞ Nc C1 Iave
y ¼ Gy  Gðy þ dyÞ  K L aCO2 ð1  eÞAdz½C 1 l¼ ½IðTÞ ð19Þ
dt RT Nc þ K n K c þ C 1 þ C 21 =K s K e þ Iave þ I2ave =K i
 C1 ð10Þ
where Nc is the concentration of nitrogen in the external medium
where G is the molar flow fate of gas (mol s1).
(g m3), Kn is the half saturation constant for nitrogen (g m3), Kc
Since the bubble residence time is short, assuming pseudo stea-
is the half saturation constant for CO2 (mol m3), Ks is the inhibition
dy state conditions, Eq. (10) can be simplified to yield:
constant for CO2 (mol m3), Iave is the average light intensity
dy KL aCO2 ð1  eÞA  (lE m2 s1), Ke is the half saturation constant for light (lE m2 -
¼ ½C 1  C 1  ð11Þ
dz G s1), and Ki is the inhibition light intensity (lE m2 s1).
Previous researchers have reported that biomass decaying rate
Using dimensionless form of Henry’s Constant of CO2 (H), liquid
due to respiration during the night is the same as the biomass
phase concentration of CO2 that is in equilibrium with the gas
decaying rate during the day, indicating that maintenance respira-
phase, C1⁄, can be related to the gas phase mole fraction of CO2 by
tion is neither stimulated nor inhibited by growth (Geider and
py Osborne, 1991; Quinn et al., 2011). Therefore, the proposed model
C 1 ¼ ð12Þ
RTH also assumed that the biomass decaying rate can account for the
Substituting from Eq. (12) into (11) and rearranging, respiration losses throughout the day and night. The model param-
eters included in Eq. (19) are described in Section 3.2.1 and 3.2.2.
dy K L aCO2 ð1  eÞA
py ¼ dz ð13Þ
RTH
 C1 G
3.2.1. Modeling the effect of light
Integrating Eq. (13) with the initial condition of y = yin at z = 0, Considering incident light intensity of Io (lE m2 s1), the aver-
the mole fraction of CO2 in the off gases, yout, of the riser of depth age light intensity within the bulk culture, Iave (lE m2 s1) can be
h, can be expressed as: estimated as follows (Molina Grima et al., 1994):
   py   
RTH in K L aCO2 ð1  eÞAp Io
yout ¼ C1 þ  C 1 exp  h ð14Þ Iave ¼ ð1  eðXDK a Þ Þ ð20Þ
p RTH GRTH XDK a
Hence, rate of transfer of CO2 from the sparging air in the riser, where D is the culture depth (m), and Ka is the light extinction coef-
Mr (mol s1) can be deduced as a function of the liquid phase CO2 ficient for biomass (m2 g1).
concentration as Ka (m2 g1) can be estimated from the following empirical equa-
    py    tion (Molina Grima et al., 1994):
RTH in K L aCO2 ð1  eÞAp
M r ¼ G yin  C1 þ  C 1 exp  h
p RTH GRTH K a ¼ 1:7356X p þ 0:0199 ð21Þ
ð15Þ
where Xp is the mass fraction of total pigment in the algae, ranging
A similar derivation can be used to derive an equation for the between 2% and 3% of the dry algal biomass (Molina Grima et al.,
rate of transfer of CO2 from the supplemental CO2 supply in the 1994).
downcomer, Ms (mol s1). Scaling laboratory scale algae growth model to predict outdoor
Since the liquid phase is assumed to be completely mixed (see biomass productivity may be inappropriate (Quinn et al., 2011) due
Supplementary material), the rate of change of liquid phase con- to the discrepancy between constant and fluctuating light intensi-
centration of CO2 can be expressed as: ties at various magnitudes. Therefore, the proposed model includes
light intensity as a function of time, incorporating the light inhibi-
dC 1
V MWCO2 ¼ Ma MWCO2 þ M r MWCO2 þ M s MWCO2  Mc ð16Þ tion effects.
dt Temperature dependence of the biomass growth is expressed in
terms of Arrhenius equation, I(T) as follows:
where V is the volume of the culture (m3), Mc is the mass rate of
consumption of CO2 by algal biomass (g s1) and MWCO2 is the IðTÞ ¼ lmax 1:066ðT C 20Þ ð22Þ
molecular weight of CO2 (=44 g mol1).
The rate of consumption of CO2 by algal biomass can be related where lmax is the maximum specific growth rate (day1) at 20 °C,
to the rate of growth of biomass as follows: and Tc is the temperature (°C).
 
1 dX 3.2.2. Modeling nitrogen uptake
Mc ¼ V ð17Þ
Y dt Since batch cultures are susceptible to nitrogen deficiency or
limitation (Rodolfi et al., 2009), the approaches suggested by Geid-
where Y is the yield of algal biomass on CO2 (g biomass g1 CO2), er et al. (1998) and Quinn et al. (2011) were followed in this study
and X is the biomass concentration (g m3). to include the effects of intercellular and extracellular nitrogen
concentration and estimate the temporal profiles for concentration
3.2. Modeling biomass growth of nitrogen Nc (g m3) in the culture media. However, reminerali-
zation of nitrogen (Geider et al., 1998) and temperature-dependent
The net growth rate of algal biomass, dX/dt, can be expressed nutrient uptake efficiencies (Packer et al., 2011) were not included
as: in the proposed model.
 B. Ketheesan, N. Nirmalakhandan / Bioresource Technology 136 (2013) 689–696 693

Uptake rate of nitrogen, U (g N g dw1 day1) is expressed in

terms of maximum specific uptake of nitrogen, Umax (g N g dw1
day1) as follows:

U ¼ U max gint;N gext;N ð23Þ

where
 
qmin
gint;N ¼ 1  ð24Þ
q
 
Nc
gext;N ¼ ð25Þ
Nc þ K n

where q is the cell quota of nitrogen in the biomass (g N g dw1) at

time t, and qmin is the minimum cell quota of nitrogen in the bio-
mass when cells cease to grow (g N g dw1).
The rate of cell quota of nitrogen in the biomass (q) can be ex-
pressed as:

dq
¼U ð26Þ
dt
By deploying initial values for Nc (g m3) and q (g N g dw1 -
day1), temporal profiles of Nc, q and U can be estimated (Quinn
et al., 2011).
In summary, the proposed model contains three coupled equa-
tions (Eqs. (15), (16), and (19)) with eight parameters relevant to
the kinetics of light, CO2, and nitrogen; and two species-specific
parameters that can predict the temporal biomass density. The
coupled differential equations, coded using a dynamic simulation
program Extend™ (Imagine That Inc.), were solved with appropri-
o
Fig. 2. Predicted ( ) vs. experimental ( ) biomass concentrations as a function of
time under laboratory conditions, at different CO2-enrichments (a–e); and overall
ate initial concentrations of C1o, Nco and, Xo. comparison (f). Species: N. salina.

3.3. Sensitivity analysis

The calibrated model was then validated with the growth data
of N. salina, obtained under different CO2-to-air ratios of 0.25%,
Sensitivity analysis procedure was performed on each of the
0.5%, 1%, and 2%. The model predictions agreed well with the tem-
model parameters to determine their significance. For each param-
poral trends in all these 4 cases as shown in Fig. 2b–e. The overall
eter, the base value established during the calibration step was
correlation coefficient (r2) between the experimental and the pre-
changed within a range of ±10% and four simulation runs were
dicted biomass concentration at different CO2-to-air ratios was
made to generate individual temporal profiles of biomass concen-
0.98 (Fig. 2f). In batch tests, biomass production with N. salina at
tration for that parameter, while all the other parameters were
CO2-to-air ratio of 2% was lower than that observed with other
kept fixed at the base value. The four biomass concentration pro-
CO2-to-air ratios of 0.25%, 0.5%, and 1% (data not shown here). As
files were compiled to form a mean profile with one standard devi-
reported by Hsueh et al. (2009), growth of Nannochloropsis sp.
ation spread. Average spread of each parameter was quantified in
can be inhibited by unutilized CO2 due to the formation of carbonic
terms of a sensitivity coefficient rDx1p defined as follows (Bernard
acid in the culture media. Therefore, an inhibitory CO2 constant
et al., 2001):
was introduced in the present model and activated only when
Z tf
1 x1ðpþDpÞ  x1ðpÞ the cultures of N. salina sparged with CO2-to-air ratios greater than
rDx1p ¼ dt ð27Þ
2%.
tf 0 x1p

where tf is the process time, x1(p+Dp) is the change in variable x1, due
to the change in the parameter value by Dp from the base value, x1p
is the value of variable x1 with respect to parameter value p.
4. Results and discussion
4.1. Model calibration and validation with N. salina under indoor
conditions
The model was calibrated using measured growth data of N. sal-
ina, cultivated under sparging with ambient air and continuous
artificial illumination. The calibration process was initiated with
eight kinetic parameters and one species-specific parameter (Ka)
obtained from literature reports to get the best-fit biomass growth
profile with r2 > 0.9 (Fig. 2a). Model parameters established by this
calibration process are listed in Table 1(a and b) along with compa-
rable literature values.
4.2. Model validation with Scenedesmus sp. in laboratory condition
The kinetic parameters established with N. salina (Table 1a)
were kept the same for Scenedesmus sp. while the species-specific
parameter was recalibrated using the growth profile of Scenedes-
mus sp. obtained under sparging with ambient air. The biomass
profile fitted during this calibration step is compared against the
measured one in Fig. 3a.
The model was validated using growth data of Scenedesmus sp.
cultivated under CO2-to-air ratios of 0.25%, 0.5%, 1%, and 3%. The
quality of fit between the predicted and measured biomass densi-
ties at these four CO2-to-air ratios was good as shown in Fig. 3b–e.
The overall correlation between the predicted and experimental
densities was statistically significant with r2 = 0.98, p < 0.001
(Fig. 3f). However, predictions under sparging with ambient air
showed slight deviations from the experimental values, probably
due to the uptake of CO2 from bicarbonate ions (Livansky, 1992)
694 B. Ketheesan, N. Nirmalakhandan / Bioresource Technology 136 (2013) 689–696

Table 1
Calibrated model parameters and comparable literature values.

Parameter Symbol Unit Value Comparable literature value

(a) Kinetic parameters
Half saturation constant for CO2 Kc mol m3 9.0E-4 8.75E-3 (Hill and Lincoln, 1981)
Half saturation constant for light Ke lE m2 s1 200 171 (Molina Grima et al., 1996)
Inhibition light intensity Ki lE m2 s1 2800 2500 (Molina Grima et al., 1994)
Maximum specific growth ratea lmax day1 2.0 1.73 (Benson et al., 2007)
Maximum specific growth rateb lmax day1 1.15 1.1 (Molina Grima et al., 1994)
Biomass decaying rate kd day1 0.06 0.05 (Yang, 2011)
Half saturation constant for nitrogen Kn g m3 0.02 0.014 (Geider et al., 1998)
Minimum nitrogen cell quota qmin g N g1 0.01 0.01 (Quinn et al., 2011)
Maximum uptake rate of nitrogen Umax g N g1d1 0.004 0.06 (Packer et al., 2011)
(b) Species-specific parameters
Inhibition constantc Ks mol m3 180 Best fit value
Extinction coefficientc Ka m2 g1 0.06 0.0752 for Nannochloropsis Oculata (Quinn et al., 2011)
Extinction coefficientd Ka m2 g1 0.0758
a
Indoor conditions.
b
Outdoor conditions.
c
For N. salina.
d
For Scenedesmus sp.

been the most common approach to identify the individual growth

determinant. As evidenced by the quality of fit in the present study,
the proposed model can be seen to be able to describe the algal
growth dynamics by incorporating the synergetic effects of dual
yield determinants.

4.3. Model calibration and validation with N. salina in outdoor

conditions

o
Fig. 3. Predicted ( ) vs. experimental ( ) biomass concentrations as a function of
time under laboratory conditions, at different CO2-enrichments (a–e); and overall
comparison (f). Species: Scenedesmus.
under carbon-limited conditions. Since CO2 tolerance of Scenedes-
mus sp. has been demonstrated in several studies as high with
CO2-to-air ratios of 5–40%, (Westerhoff et al., 2010), inhibitory ef-
fect of CO2 was not considered in this validation process.
In the previous study (Ketheesan and Nirmalakhandan, 2012), a
notable increase in algal growth was observed in the tests with
CO2-enrichment (0.5–3%) compared to that in the test with ambi-
ent air. However, growth of Scenedesmus sp. seemed to be less
influenced by elevated CO2-to-air ratios (0.5–3%) during declining
growth phase as a consequence of co-limitation by light and nitro-
gen (Westerhoff et al., 2010). Concerning the multiple limiting
nutrient effects in algal cultivation, specific growth rate estima-
tions or stoichiometric evaluations (Hill and Lincoln, 1981) have
The model was calibrated (Fig. 4a) against the growth data of N.
salina cultivated outdoors with on-demand supply of CO2 (pH con-
trol at 7.5). During this outdoor cultivation period (September 3,
2011 to September 22, 2011), diurnal light intensity incident on
the raceway was ranging between 60 and 1100 lE m2 s1. As
shown in Fig. 4b and c, the calibrated model was validated using
temporal growth profiles of N. salina cultivated under sparging
with ambient air and with supplementary CO2 (pH control at
7.5). During this cultivation period (October 31, 2011 to November
22, 2011), diurnal light intensity incident on the raceway ranged
between 30 and 600 lE m2 s1. The predicted biomass densities
agreed well with the measured ones with r2 of 0.98 (Fig. 4d).
Photoadaptation is an imperative phenomenon in outdoor algal
cultivation under fluctuating light intensities. In photoadaptation
process, unicellular algae physiologically adjust its photosynthetic
apparatus to a given light intensity, thus optimizing the photosyn-
thetic capacity of the cell (Falkowski, 1984). Although photoadap-
tation can be described by first-order kinetics (Falkowski, 1984),
modeling photoadaptation is quite complicated (Sukenik et al.,
1991) due to interactions by temporal variations in the light inten-
sity and light attenuation effects, which may require additional
model parameters. Also, several literature reports have indicated
that the respiration losses become significant in outdoor cultures
(2–10% of biomass production) compared to continuous illumina-
tion conditions as a result of biomass losses at night and dark res-
piration caused by light attenuation during day time (Grobbelaar
and Soeder, 1985; Sukenik et al., 1991). Therefore, the maximum
specific growth rate (lmax) of N. salina under constant light inten-
sity (2 day1) had to be adjusted as 1.15 day1 to calibrate the
model for outdoor conditions.
The use of the proposed model can be illustrated by the follow-
ing case; on-demand supply of CO2 via pH control (Fig. 4b) did not
show any significant increase in biomass production compared to
sparging with ambient air (Fig. 4c) though both reactors were
operated simultaneously under identical light intensity and
temperature conditions. Since light intensity was the prominent
 B. Ketheesan, N. Nirmalakhandan / Bioresource Technology 136 (2013) 689–696 695

Fig. 5. Sensitivity of predicted biomass concentration to ±10% variations in the

three most sensitive parameters: (a) Kn; (b) Ke; and (c) lmax. Species: N. salina.

CO2 utilization efficiency or CO2 fixation rate instead (Westerhoff

o
Fig. 4. Predicted ( ) vs. experimental ( ) biomass concentrations as a function of
time under field conditions, with and without pH-control (a–c); and overall
comparison (d). Species: N. salina.
growth determinant for biomass growth, supply of carbon via
ambient air alone was adequate to sustain the photosynthetic car-
bon needs in that case, which is well accounted for by the proposed
model.
4.4. Sensitivity analysis
The sensitivity analysis procedure was conducted for the eight
kinetic model parameters (Kc, kd, Ke, lmax, qmin, Ki, Kn, and Umax)
and the two species-specific parameters (Ka and Ks) where, the im-
pact of ±10% variation in the value of each parameter was simu-
lated. Growth of N. salina under sparging with ambient air under
indoor condition was simulated for this analysis. Based on the sen-
et al., 2010). Based on the model’s ability to predict well the
growth under various CO2 enrichments, it is proposed that the
model could be used to optimize the CO2 enrichment to maximize
net energy production costs. As an illustration, simulated cost anal-
ysis is presented here to evaluate the benefit of CO2 enrichment.
According to Xu et al. (2010), cost of CO2 for microalgal cultivation
is dependent on the source of production of CO2 such as ethanol
facilities, hydrogen and ammonia production, gas-processing
plants, and fossil power plants. In this illustration, cost of CO2 is
considered as 12 USD (ton CO2)1, assuming CO2 is obtained from
ethanol production facilities (Xu et al., 2010; Kumar et al., 2011). It
is assumed that the calorific values of algal biomass can be con-
verted into electricity via an internal combustion engine/alternator
with an efficiency of 60%; and that the price of electricity is
11.58 cents kW h1 (EIA, 2011). Based on the above, the net cost
of algal-produced electricity can be estimated by deducting the
cost of CO2 from the cost of electricity produced at different
CO2-air ratios in the sparging gas.
0.8 100
Case 1: 0.014 g/cu m
0.6 Case 2: 0.028 g/cu m
CO2 utilization efficiency [%]

80
Net costs [US cents/kW-hr]

sitivity coefficients calculated from these simulations, maximum 0.4

specific growth rate, lmax (rxDp ¼ 0:13) was found to be the most
sensitized parameter followed by the half saturation constant for 60
0.2
light, Ke (rxDp ¼ 0:11) and the half saturation constant for nitrogen,
Kn (rxDp ¼ 0:08). The deviations of the simulated biomass growth 0.0
40
profiles in these three cases relative to the base profile are illus-
-0.2
trated in Figs 5a–c.
20
-0.4
4.5. Application of the proposed model: optimizing CO2 enrichment
-0.6 0
0 0.2 0.4 0.6 0.8 1
Several approaches have been suggested in literature to opti-
mize the level of CO2 enrichment in gas-sparged photobioreactors. Fig. 6. Simulation of net cost (US cents) and CO2 utilization efficiency (%) as a
While some studies have optimized CO2 enrichment with respect function of CO2-air ratio for N. salina, for two cases: Case 1 – light 200 lE m2 s1;
to biomass production (Hsueh et al., 2009), few have proposed 0.014 g m3; Case 2 – light 200 lE m2 s1; 0.028 g m3.
696 B. Ketheesan, N. Nirmalakhandan / Bioresource Technology 136 (2013) 689–696
Fig. 6 shows the simulated results of net benefit (US cents) and
CO2 utilization efficiency (%) in two different cases, as a function of
CO2-to-air ratios. As CO2-to-air ratio is increased, CO2 utilization
efficiency decreased continuously, whereas, the net cost increased
initially and then declined with further increase in CO2-to-air ra-
tios. For instance, the use of CEA could only be beneficial in case
2 if CO2-to-air ratio could be maintained within 0.1–0.2% based
on net benefit. In fact, maximum cost benefit was observed when
the CO2-to-air ratio ranged between 0.12% and 0.15%. Thus, the
proposed model can be used to regulate the input of CO2 in re-
sponse to temporal operational variables and culture conditions
for optimum CO2 usage.
5. Conclusion
The use of airlift column in the open raceway for the supply and
transfer of CO2 is modeled, integrating it with a model for predict-
ing biomass growth, incorporating the parameters such as light,
CO2, nitrogen and temperature. Predictions of biomass concentra-
tions by the proposed model agreed well with the temporal trend
of the experimental data, with r2 ranging from 0.96 to 0.98,
p < 0.001. Based on model simulations, the proposed model could
prove useful in scaling up and optimizing the input of CO2 in the
proposed airlift-raceway configuration in response to temporal
operational variables and culture conditions.
Acknowledgements
This study was supported in part by a Grant from the DOE Na-
tional Alliance for Advanced Biofuels and Bioproducts (NAABB), a
Grant from the US Air Force Research Laboratory (AFRL), by the
NSF Engineering Research Center: RenuWIT, and by the Ed & Har-
old Foreman Endowed Chair.
Appendix A. Supplementary material
Supplementary material associated with this article can be
found, in the online version, at http://dx.doi.org/10.1016/
j.biortech.2013.02.028.
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