Journal of Biotechnology 129 (2007) 439445

Biofixation of carbon dioxide by Spirulina sp. and Scenedesmus

obliquus cultivated in a three-stage serial tubular photobioreactor
Michele Greque de Morais, Jorge Alberto Vieira Costa
Laboratory of Biochemical Engineering, Department of Chemistry, Federal University Foundation of Rio Grande,
P.O. Box 474, Rio Grande 96201-900, RS, Brazil
Received 28 August 2006; received in revised form 4 January 2007; accepted 12 January 2007

Abstract
The increase in the concentration of atmospheric carbon dioxide is considered to be one of the main causes of global warming. As estimated
by the Intergovernmental Panel on Climate Change (IPCC) criteria, about 1015% of the gases emitted from the combustion coal being in the
form of carbon dioxide. Microalgae and cyanobacteria can contribute to the reduction of atmospheric carbon dioxide by using this gas as carbon
source. We cultivated the Scenedesmus obliquus and Spirulina sp. at 30 C in a temperature-controlled three-stage serial tubular photobioreactor
and determined the resistance of these organisms to limitation and excess of carbon dioxide and the capacity of the system to fix this greenhouse
gas. After 5 days of cultivation under conditions of carbon limitation both organisms showed cell death. Spirulina sp. presenting better results for
all parameters than S. obliquus. For Spirulina sp. the maximum specific growth rate and maximum productivity was 0.44 d1 , 0.22 g L1 d1 , both
with 6% (v/v) carbon dioxide and maximum cellular concentration was 3.50 g L1 with 12% (v/v) carbon dioxide. Maximum daily carbon dioxide
biofixation was 53.29% for 6% (v/v) carbon dioxide and 45.61% for 12% carbon dioxide to Spirulina sp. corresponding values for S. obliquus
being 28.08% for 6% (v/v) carbon dioxide and 13.56% for 12% (v/v) carbon dioxide. The highest mean carbon dioxide fixation rates value was
37.9% to Spirulina sp. in the 6% carbon dioxide runs.
2007 Elsevier B.V. All rights reserved.

Keywords: Carbon dioxide fixation; Chlorophyta; Cyanobacteria; Global warming; Greenhouse gases; Microalgae

1. Introduction dioxide, the principal greenhouse gas. According to the French

The growth in human population has stimulated the search
for alternative food sources and new ecological technologies.
Microalgae and cyanobacteria use solar light as their main
source of energy, possess the potential for high productivity, are
tolerant to alterations in environmental conditions and they can
be cultivated in areas which are unstable for agriculture (Costa
et al., 2000). The cultivation conditions of these organisms can
be manipulated to induce the production of proteins, fatty-acids,
vitamin A, minerals, pigments and other bio-compounds and
their biomass can be used as a dietary supplement for humans
and animals, including for aquaculture (Ono and Cuello,
2004).
Photosynthetic microorganisms use inorganic carbon for
growth and hence can be used for the biofixation of carbon
Corresponding author. Tel.: +55 53 3233 8653; fax: +55 53 3233 8745.
E-mail address: dqmjorge@furg.br (J.A.V. Costa).
National Center for Scientific Research (CNRS), the level
of atmospheric carbon dioxide has historically been between
180 and 260 ppm but during the last 100 years the atmo-
spheric concentration of this gas has risen to between 260 and
380 ppm (Siegenthaler et al., 2005), mainly due to burning fossil-
fuels associated with increased population and industrialization
(Chang and Yang, 2003). Coupling the cultivation of photosyn-
thetic microorganisms with the biofixation of carbon dioxide has
the potential not only to reduce the costs of culture media for
growing such organisms on an industrial scale but also to offset
carbon emissions (Beneman and Hughes, 1997).
Photosynthetic microorganisms are normally cultivated in
open raceway tanks using natural or artificial light but this
requires large cultivation areas and suffers from various
disadvantages, including difficulty in controlling cultivation
conditions (Costa et al., 2006), evaporation of the cultivation
medium and reduced light intensity with increased depth. An
alternative is the use of tubular photobioreactors, but Luo et al.
(2003) have pointed out that the configuration of such reactors

0168-1656/$ see front matter 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.jbiotec.2007.01.009
440 M.G. de Morais, J.A.V. Costa / Journal of Biotechnology 129 (2007) 439445
is one of the main factors controlling the biomass productivity
of photosynthetic cultures grown under these conditions.
Appropriately designed photobioreactors can reduce the cul-
tivation area by distributing photosynthetic organisms vertically,
vertical tubular photobioreactors also increasing the carbon
dioxide residence time in the cultivation medium and, con-
sequently, the carbon dioxide utilization efficiency (Ono and
Cuello, 2004). Cultivations can also be carried out in serial pho-
tobioreactors in which unused effluent carbon dioxide from one
photobioreactor is fed into another photobioreactor.
The objective of the work described in this paper was to culti-
vate the photosynthetic microorganisms Scenedesmus obliquus
and Spirulina sp. in serial tubular photobioreactors and deter-
mine carbon dioxide fixation and their resistance to carbon
dioxide limitation and excess.
2. Material and methods
2.1. Microorganisms and cultivation media
The Spirulina sp. (Cyanobacteria, Oscillatoriales) and S.
obliquus (Chlorophyta, Chlorophyceae) (de Morais and Costa,
2007) were from stock cultures kept in our laboratory. We
used carbon-free media for the maintenance and cultivation
of both organisms, modified Zarrouk medium (de Morais and
Costa, 2007; Zarrouk, 1966) for Spirulina sp. and MC medium
(Watanabe, 1960) for S. obliquus. For the experiments, inocu-
lum of both organisms were acclimatized to carbon dioxide by
maintaining them for 7 days under air mixed with 1% (v/v) added
carbon dioxide.
2.2. Photobioreactors and cultivation conditions
Pure cultures of Spirulina sp. and S. obliquus were individ-
ually cultivated in three 2 L (1.8 L working volume) column
photobioreactors (CPBR) connected in series (Fig. 1) and
labeled consecutively as CPBR1, CPBR2 and CPBR3. Agitation
and aeration were accomplished using air from a compressor
Fig. 1. Three-stage serial photobioreactors scheme. All measurements in mil-
limetres.
and a sintered sparger, the effluent air (with or without car-
bon dioxide, see below) from CPBR1 being fed to the sparger
in CPBR2 and the effluent from this photobioreactor being
fed to CPBR3. The photobioreactors were maintaining in a
30 C growth chamber under a 12 h dark/light photoperiod with
3200 lx of illumination provided by 40 W daylight-type fluo-
rescent lamps (General Electric, Brazil) during the light period
(Reichert et al., 2006).
For experiments without carbon dioxide filter-sterilized air
was bubbled through 1N sodium hydroxide and then twice
through water to remove the approximately 0.038% (v/v) present
in air. In the experiments with carbon dioxide filter-sterilized
carbon dioxide (White Martins, Brazil) was added to the air
entering CPBR1 at a rate of 0.3 vvm for 15 min every 2 h during
the 12 h light period such that the final carbon dioxide concen-
tration in the media was 6% or 12% (v/v). The initial biomass
concentration in all runs was 0.15 g L1 and all runs lasted for
21 days.
2.3. Analytic determinations
Triplicate samples of culture media were collected asepti-
cally at 24 h intervals and pH determined with a Q400H digital
pH meter (Quimis, Brazil) and biomass concentration (X, g L1 )
calculated by measuring the optic density at 670 nm (Costa et
al., 2002) using a 700 Plus spectrophotometer (Femto, Brazil)
and a calibration curve of optic density versus dry biomass
(Reichert et al., 2006). Biomass carbon content was measured
using a Perkin-Elmer 2400 CHNS (carbon, hydrogen, nitrogen
and sulfur) element analyzer calibrated to the 100% value using
a certified cystine standard (Perkin-Elmer, USA).
2.4. Kinetic parameters
Biomass (X) values and exponential regression were used to
calculate the maximum specific growth rate (max , d1 ) dur-
ing the logarithmic phase and maximum specific death rate (k,
d1 ) during the decline phase (Bailey and Ollis, 1986). The
doubling time (td , d) was calculated as td = ln 2(max )1 . The
maximum biomass concentration achieved in a photobioreac-
tor was designated Xmax (g L1 ). Productivity (P, g L1 d1 )
was obtained using the equation P = (Xt X0 )(t t0 )1 where
Xt is the biomass concentration (g L1 ) at t (d) and X0 is
the biomass concentration at inoculation (t0 ) (Schmidell et al.,
2001). Maximum productivity during cultivation was designated
Pmax (g L1 d1 ).
2.5. Carbon dioxide xation
Carbon dioxide fixation (F, g) by each organism was calcu-
lated from the CHNS biomass carbon content values.
The accumulation of fixed carbon dioxide (FA, g) in each
of the three individual photobioreactors which made up the
photobioreactor was calculated as FA = (Xt X0 )mcbm Vp (mCO2
mC 1 ), where FA can represent FA1, FA2 or FA3, where FA1,
FA2 and FA3 represent the accumulation of fixed carbon dioxide
in CPBR1, CPBR2 and CPBR3, respectively; Xt is the biomass
 M.G. de Morais, J.A.V. Costa / Journal of Biotechnology 129 (2007) 439445 441

Table 1
Maximum specific growth rate (max , d1 ); maximum biomass concentration (Xmax , g L1 ); maximum productivity (Pmax , g L1 d1 ) for Spirulina sp. and S.
obliquus growing in the three different photobioreactors (CPBR) which constituted the three-stage serial photobioreactor, with and without carbon dioxide as the
sole carbon source
Carbon dioxide level (%, v/v)

Microalga and CPBR 0 6 12

max Xmax Pmax max Xmax Pmax max Xmax Pmax

Spirulina sp.
CPBR1 0.33 a 0.82 a 0.04 a 0.44 3.40 c 0.20 0.33 a 3.38 cf 0.17 e
CPBR2 0.33 a 0.85 a 0.14 b 0.27 abe 3.00 d 0.22 0.32 abe 3.20 0.17 e
CPBR3 0.29 ab 0.80 a 0.14 b 0.27 abe 2.90 d 0.19 0.29 ab 3.50 cf 0.18
S. obliquus
CPBR1 0.15 c 0.31 b 0.04 a 0.22 cf 1.56 0.10 d 0.22 cf 1.80 e 0.14 b
CPBR2 0.10 cd 0.27 b 0.06 ac 0.19 cfg 1.81 e 0.10 d 0.14 cdgh 1.50 g 0.10 d
CPBR3 0.07 d 0.24 b 0.05 a 0.19 cfg 1.36 0.07 c 0.14 cdgh 1.50 g 0.09 d

The air and, when present, carbon dioxide were fed serially into the photobioreactors starting with CPBR1. Values with the same letters in the same parameters
indicate that the values did not differ by the Tukey test at p 0.10.

concentration (g L1 ) at t (d) and X0 is the biomass concentra-
tion at inoculation (t0 ); mcbm is the fraction of carbon in the
biomass (g g1 ); Vp (L) is the volume of culture medium in a
specific photobioreactor; mCO2 (g mol1 ) is the molar mass of
carbon dioxide; mC (g mol1 ) the molar mass of carbon.
The total daily carbon dioxide fixation per gram of
carbon dioxide added to the medium (TF, g g1 ) was calcu-
lated for three-stage serial tubular photobioreactor as TF =
(FA1(t+1) + FA2(t+1) + FA3(t+1) ) (FA1t + FA2t + FA3t ) m24 h 1 ,
where FA1(t+1) , FA2(t+1) and FA3(t+1) represent the fixed carbon
dioxide accumulated in CPBR1, CPBR2 and CPBR3 at t + 1
(d), respectively. FA1t , FA2t and FA3t represent the fixed
carbon dioxide accumulated in CPBR1, CPBR2 and CPBR3 at
time t (d); m24 h represents the mass of carbon dioxide (g) added
in 24 h. The maximum total daily carbon dioxide fixation was
designated MDF.
The mean carbon dioxide fixation in grams of total carbon
dioxide fixed per gram of total carbon dioxide added to the
media (MF, g g1 ) for three-stage serial tubular photobiore-
actor was calculated as MF = (FA1(t+1) + FA2(t+1) + FA3(t+1) )
(FA1t + FA2t + FA3t ) [m24 h (t + 1)]1 , where FA1(t+1) , FA2(t+1)
and FA3(t+1) represent the fixed carbon dioxide accumulated
in CPBR1, CPBR2 and CPBR3 at t + 1 (d); FA1t , FA2t and
FA3t represent the fixed carbon dioxide accumulated in CPBR1,
CPBR2 and CPBR3 at time t = 1 d; m24 h represents the mass (g)
of carbon dioxide added in 24 h.
2.6. Statistical analysis
The experimental results were evaluated by comparison of
the growth curves and analysis of variance (ANOVA) of the
kinetic parameters, significance being tested by the Tukey test
at p 0.10.
3. Results
The maximum specific growth rate and maximum biomass
concentration values for S. obliquus and Spirulina sp. growing
in the three-stage serial tubular photobioreactors in the presence
of three different concentrations of carbon dioxide are shown in
Table 1 and the growth curves of biomass versus time are shown
in Figs. 24.
In the absence of carbon dioxide Spirulina sp. runs biomass
increased until day 5 of cultivation (Fig. 2a), with no signif-

Fig. 2. Time course of biomass concentration for Spirulina sp. (a) and Scenedesmus obliquus (b) cultive in the absence of carbon dioxide in three-stage serial tubular
photobioreactor, where CPBR1 (), CPBR2 () and CPBR3 (+).
442 M.G. de Morais, J.A.V. Costa / Journal of Biotechnology 129 (2007) 439445

Fig. 3. Time course of biomass concentration for Spirulina sp. (a) and S. obliquus (b) using air supplemented with 6% (v/v) of carbon dioxide added to three-stage
serial tubular photobioreactor, where CPBR1 (), CPBR2 () and CPBR3 (+).

icant difference (p > 0.6004) between the maximum biomass
concentration values for the three photobioreactors (Table 1).
Under these conditions the Spirulina sp. Xmax values (g L1 )
were significantly smaller (p < 0.0002) than those for the 6%
and 12% carbon dioxide runs (Table 1) but there was no sig-
nificant difference between the maximum productivity (Pmax ,
g L1 d1 ) of Spirulina sp. in the different photobioreactors,
with CPBR1 giving a Pmax value of 0.04 g L1 d1 and CPBR2
and CPBR3 both giving 0.14 g L1 d1 . The max values were
similar for all three photobioreactors but after 5 days cultivation
in the absence of carbon dioxide the Spirulina sp. biomass
values fell abruptly and the specifics death rates (k, d1 ) were
0.36, 0.22 and 0.43 d1 for CPBR1, CPBR2 and CPBR3,
respectively, with the stationary phase being reached after
10 days.
At the same carbon dioxide level, the S. obliquus biomass
concentration also increased until day 5 (Fig. 2b) but the Xmax
values were significantly smaller (p < 0.0002) than those for
Spirulina sp. and S. obliquus max values were also signifi-
cantly lower than the corresponding values for Spirulina sp.
and there was a significant difference (p = 0.0015) between
CPBR1 and CPBR3 (Table 1). Under the same conditions, the
S. obliquus Pmax value for CPBR1 was 0.04 g L1 d1 , the same
as for Spirulina sp., but was only 0.06 g L1 d1 for CPBR2 and
0.05 g L1 d1 for CPBR3, lower than the corresponding values
for Spirulina sp.
In the Spirulina sp. runs with 6% carbon dioxide (Fig. 3a)
the Xmax values were high and there were significant dif-
ferences (p 0.0002) between the different photobioreactors
(Table 1), with the highest value being obtained in CPBR1 which
received the largest amount of carbon dioxide because it was
the first in the series (Table 1). During the 21 days of culti-
vation the concentration of Spirulina sp. biomass increased in
the three serial photobioreactors and the Pmax values for Spir-
ulina sp. in CPBR1, CPBR2 and CPBR3 were 0.20, 0.22 and
0.19 g L1 d1 , respectively. The max values for Spirulina sp.
cultivated in CPBR1 was significantly higher (p < 0.0002) than
that achieved in CPBR2 and CPBR3 which both showed the
same max values (Table 1).
The S. obliquus runs with 6% carbon dioxide (Fig. 3b) showed
significantly higher (p < 0.0002) Xmax values in CPBR2 than in
CPBR1 and CPBR3 (Table 1). The S. obliquus Pmax values were
0.10 g L1 d1 in CPBR1 and 0.07 g L1 d1 in CPBR2 and
CPBR3, significantly lower (p < 0.0002) than the corresponding
values for Spirulina sp. The max value for S. obliquus was
highest in CPBR1, although there was no statistically significant

Fig. 4. Time course of biomass concentration for Spirulina sp. (a) and S. obliquus (b) using air supplemented with 12% (v/v) carbon dioxide added three-stage serial
tubular photobioreactor, where CPBR1 (), CPBR2 () and CPBR3 (+).
 M.G. de Morais, J.A.V. Costa / Journal of Biotechnology 129 (2007) 439445
Fig. 5. Mean carbon dioxide fixation in three serial photobioreactors in the
presence of 6% (v/v) carbon dioxide to Spirulina sp. () and S. obliquus ()
and with 12% (v/v) carbon dioxide to Spirulina sp. (), S. obliquus ().
difference between this value and the corresponding values for
CPBR2 and CPBR3.
With 12% carbon dioxide the Spirulina sp. Xmax and max
values were significantly higher (p < 0.0002) than those for S.
obliquus (Table 1, Fig. 4a). For S. obliquus cultivated using the
same carbon dioxide concentration (Fig. 4b) Xmax was signifi-
cantly higher (p < 0.0002) in CPBR1 as compared with CPBR2
and CPBR3, which both showed the same values (Table 1).
The mean carbon dioxide biofixation rates (MF) in the three
serial photobioreactors for Spirulina sp. and S. obliquus growing
on 6% and 12% carbon dioxide are shown in Fig. 5. In CPBR2
and CPBR3 the cyanobacterium Spirulina sp. presented signif-
icantly higher (p = 0.0002) MF values than did the microalgae
S. obliquus, but for CPBR1 there was no significant difference
between the MF values for Spirulina sp. growing on 6% and
12% carbon dioxide and S. obliquus growing on 6% carbon
dioxide (data not shown). The highest Spirulina sp. MF values
(27.1437.9%) occurred in the 6% carbon dioxide runs, with
the lowest mean carbon dioxide biofixation rates values in these
runs being higher than the highest values in all other runs. The
MF value for S. obliquus cultivated with 6% carbon dioxide was
7.4013.45%. In the 12% carbon dioxide runs the Spirulina sp.
MF value was 6.7017.06% while for S. obliquus it varied from
4.39% to 8.63%. Maximum daily carbon dioxide biofixation
(MDF) for Spirulina sp. was 53.29% on day 7 and 45.61% on day
14 when growing on 6% and 12% carbon dioxide, respectively,
while the corresponding values for S. obliquus were significantly
lower (p 0.0541) at 28.08% on day 8 and 13.56% on day 9
when growing on 6% and 12% carbon dioxide, respectively. In
the absence of carbon dioxide biomass production ceased after
5 days of cultivation.
The pH values in all three photobioreactors were very simi-
lar for both Spirulina sp. and S. obliquus at both carbon dioxide
concentrations. For Spirulina sp. the average pH of the three
serial photobioreactors was higher (9.70 0.20 to 11.71 0.50)
in the absence of carbon dioxide than in the 6% (initial pH
7.16 0.16, then 9.33 0.05 after 24 h and 10.22 0.61 in
443
21 days) or 12% (initial pH 7.08 0.29 rising to 8.76 0.15
in 21 days) carbon dioxide runs. When S. obliquus was culti-
vated without carbon dioxide the initial pH was 6.95 0.03 but
increased to pH 9.18 0.26 after 24 h and reached a maximum
of about pH 10.03 0.64 after 15 days, similar to the 6% (initial
pH 5.40 0.01 rising to 8.62 0.39 in 21 days) and 12% (initial
pH 5.33 0.05 rising to 9.09 0.77 in 21 days) carbon dioxide
runs.
These results may have been due to the fact that in the absence
of carbon dioxide there was no formation of carbonic acid and
no abrupt decreases in pH. However, when carbon dioxide was
present the carbonic acid formed reduced the pH of the media
but this began to increase as the carbon dioxide was metabo-
lized by the microorganisms, which had been acclimatized to
1% carbon dioxide. Although no attempt was made to maintain
the organisms at the most appropriate pH for optimum growth
this did not appear to have any adverse effects on the growth,
indicating that both these organisms are resistant to variations
in pH.
4. Discussion
When fossil fuels are burned they produce about 12% of
carbon dioxide (Lee et al., 2002) and it has been reported that
cultivation of microalga could fix carbon dioxide equivalent to
500 MW of energy (Kadam, 2002). Our results show that both
Spirulina sp. and S. obliquus can grow in media supplied with
air containing this concentration of carbon dioxide, indicating
that these organisms could be used to sequester the carbon diox-
ide produced by combustion gases from thermoelectric power
stations.
The maximum mean carbon dioxide biofixation rates values
for Spirulina sp. were recorded on day 7 with 6% carbon dioxide
and day 14 with 12% carbon dioxide, while for S. obliquus the
maximum MF values occurred on day 8 with 6% carbon dioxide
and day 9 with 12% carbon dioxide.
The growth rate of the diatom Chaetoceros wighamii (Araujo
and Garcia, 2005) and the microalga Tetraselmis (Olaizola et
al., 1991) increased when carbon dioxide was added to their
growth medium, indicating that carbon dioxide can be a limiting
factor for the growth of a variety of different photosynthetic
microorganisms. Chang and Yang (2003) cultivated Chlorella
strains NTU-H15 and NTU-H25 and found that the addition of
carbon dioxide stimulated the growth of the cells and resulted in
high productivity of 0.31 g L1 d1 , with both strains showing
the best results with 5% carbon dioxide.
Sung et al. (1999) grew Chlorella strain KR-1 and reported
different productivity with different levels of carbon dioxide
(10%, 1.1 g L1 d1 ; 30%, 0.8 g L1 d1 ; 50%, 0.6 g L1 d1
and 70%, 0.1 g L1 d1 ), although these cultivations were not
conducted in series. Hanaga et al. (1992) grew Scenedesmus and
Chlorella reported that both organisms presented a productivity
of 0.15 g L1 d1 with 10% carbon dioxide and 0.18 g L1 d1
with 40% carbon dioxide. The concentration of carbon dioxide
in a culture media should never be less that required for the max-
imum growth rate of the culture but should not be so high as to
exceed the maximum tolerated by a particular organism, thus
444 M.G. de Morais, J.A.V. Costa / Journal of Biotechnology 129 (2007) 439445
avoiding reduced fixation and/or loss to the atmosphere (Cheng
et al., 2006).
Although the two organisms behaved differently regarding
the carbon fixation, there was no obvious lag phase in any of
the runs (Figs. 24), possibly because both the Spirulina sp. and
S. obliquus cultures used for the inoculum had been preadapted
to carbon dioxide. Travieso et al. (2001) preadapted Spirulina
platensis cultures to carbon dioxide but still found a lag phase of
2 days. Yun et al. (1997) reported that cultures of the microalga
Chlorella vulgaris which had been pre-adapted to 5% carbon
dioxide grew better in the presence of 15% carbon dioxide as
compared to unadapted cultures and Lee et al. (2002) found a
prolonged lag phase in unadapted Chlorella cultures when the
carbon dioxide concentration was increased from 10% to 70%.
For Spirulina sp. and S. obliquus the highest average pH
were 11.71 0.50 and 10.03 0.64, respectively, in the absence
of carbon dioxide, and the lower value pH were 7.08 0.29
and 5.33 0.05, respectively, in the 12% carbon dioxide runs.
Regarding pH, the microalga Chlorella strain KR-1 cultivated
with carbon dioxide maintained constant growth at pH 4.2 but
was completely inhibited at pH 3.5 (Sung et al., 1999) but
Chlorella strain ZY-1 grew well at pH values of 4.06.0 (Yue
and Chen, 2005) while the growth of Chlorococcum littorale
was unaffected by changes in pH (Kodama et al., 1993). When
Chlorella vulgaris was illuminated and able to perform photo-
synthesis the culture media was maintained between pH 10.0
and 10.5 but this dropped to around 8.5 in the dark when there
was respiration but no photosynthesis (Cheng et al., 2006).
Several factors influence productivity, including the effi-
ciency of the photosynthetic pigments in converting light energy
into energy for biomass production, the accumulation of oxy-
gen produced by photosynthesis, biomass consumption during
respiration, insufficient carbon dioxide transfer, nutrient short-
fall and photoinhibition (Stewart and Hessami, 2005; Degen
et al., 2001). Factors such as light intensity, cultivation den-
sity, photobioreactor type and carbon dioxide concentration can
be limiting conditions influencing photosynthesis and inhibit-
ing carbon dioxide biofixation (Cheng et al., 2006). In dense
cultures of photosynthetic microorganisms shading effects can
occur which reduce the level of light reaching individual cells
and thus prevent carbon dioxide fixation, although this can be
avoided by the periodic removal of cells (Costa et al., 2004). The
advantage of this system in serial photobioreactors being higher
levels of carbon dioxide retention with lower levels of this gas
in the final gaseous effluent vented to the atmosphere, showed
53.29% and 45.61% for maximum daily carbon dioxide biofixa-
tion when growing on 6% and 12% carbon dioxide, respectively.
5. Conclusions
This research shows the high carbon dioxide biofixation
potential of the cyanobacteria Spirulina sp. as indicated by its
higher kinetic parameters and mean and maximum daily fixa-
tion rates as compared to the microalga S. obliquus. For Spirulina
sp. the highest biomass concentration was 3.40 g L1 with 6%
carbon dioxide and 3.50 g L1 with 12% carbon dioxide, while
the highest maximum specific growth rate was 0.44 d1 and
the highest maximum productivity was 0.22 g L1 d1 , both in
the presence of 6% carbon dioxide. Maximum daily carbon
dioxide biofixation for Spirulina sp. was 53.29% and 45.61%
when growing on 6% and 12% carbon dioxide, respectively,
while the corresponding values for S. obliquus were significantly
(p 0.0541) lower at 28.08% and 13.56% when growing on 6%
and 12% carbon dioxide, respectively. In the absence of carbon
dioxide biomass production ceased after 5 days of cultivation.
The highest mean carbon dioxide fixation rates value was 37.9%
to Spirulina sp. in the 6% carbon dioxide runs.
Acknowledgements
The authors thank the Centrais Eletricas Brasileiras S.A.
(ELETROBRAS) and the Companhia de Geracao Termica de
Energia Eletrica (CGTEE) for financially supporting this work.
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