Bioresource Technology 209 (2016) 351–359

Contents lists available at ScienceDirect

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

A new technology of CO2 supplementary for microalgae cultivation

on large scale – A spraying absorption tower coupled with an outdoor
open runway pond
Chun-Dan Zhang a, Wei Li a, Yun-Hai Shi a,⇑, Yuan-Guang Li b, Jian-Ke Huang b, Hong-Xia Li a
a
Research Centre of Chemical Engineering, East China University of Science and Technology, Shanghai 200237, China
b
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China

h i g h l i g h t s

A spray tower designed as a novel CO2 supply device was studied in algae cultivation.

Chlorella pyrenoidosa can be cultivated using pure CO2 in this new method.

Microalgae productivity, pH and CO2 consumption were investigated.

Mean fixation ratio of CO2 by microalgae in ORWP coupled with spraying tower was 50%.

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

Article history: An effective CO2 supply system of a spraying absorption tower combined with an outdoor ORWP (open
Received 4 January 2016 raceway pond) for microalgae photoautotrophic cultivation is developed in this paper. The microalgae
Received in revised form 26 February 2016 yield, productivity and CO2 fixation efficiency were investigated, and compared with those of bubbling
Accepted 1 March 2016
method. The maximum yield and productivity of biomass were achieved 0.927 g L1 and
Available online 7 March 2016
0.114 g L1 day1, respectively. The fixation efficiency of CO2 by microalgae with the spraying tower
reached 50%, whereas only 11.17% for bubbling method. Pure CO2 can be used in the spraying absorption
Keywords:
tower, and the flow rate was only about one third of the bubbling cultivation. It shows that this new
Chlorella
CO2 biological fixation
method of quantifiable control CO2 supply can meet the requirements of the growth of microalgae cul-
Spraying absorption tower tivation on large-scale.
High CO2 concentration Ó 2016 Elsevier Ltd. All rights reserved.

1. Introduction and strong adaptability to environments (Keffer and Kleinheinz,

Cultivation of microalgae on large scale, which can provide
clean and renewable biofuels, has been extensively investigated
worldwide in recent years. Microalgae usually contain about 50%
of carbon, which means 1.83 (calculated as 0.50 divided by the car-
bon content of CO2 molecules) tons of CO2 needed to produce one
ton of microalgae through photosynthesis (Li and Luo, 2013).
Therefore, it could be a feasible method by using microalgae to
diminish emissions of concentrative CO2 exhaust gases from fossil
flue-fired power plants (Watanabe and Ohmura, 1992). On the
other hand, valuable products from microalgae, such as proteins,
starch, vitamins (Chae et al., 2005) could be obtained simultane-
ously through photosynthesis with high rate, fast reproduction
⇑ Corresponding author. Tel.: +86 2164252355; fax: +86 2164252460.
E-mail addresses: shi_yunhai@sina.com, shi_yunhai@ecust.edu.cn, shi_yunhai@
hotmail.com (Y.-H. Shi).
2002).
The fixation efficiency and cost of CO2 must be considered dur-
ing the microalgae cultivation process. The conventional apparatus
of photo-autotrophic microalgae cultivation is operated with the
pipeline gas supply directly immersed into the microalgae cultiva-
tion solution to bubble for carbon complementary (Li and Luo,
2013; Morais and Costa, 2007a). This operation mode has been
given sufficient attention with simple equipment, convenient oper-
ation, less capital and operational cost (Chiaramonti et al., 2013).
However, during the process of algae cultivation, this method does
not consider low CO2 fixation efficiency and hypersensitivity of
algae under the high CO2 concentration and low pH value environ-
ment (Kurano et al., 1995). The concentration of CO2 in the exhaust
gas usually reaches up to 20–30% (vol.) with high temperature (80–
150 °C) and a small amount of toxic gases, such as SOx, NOx, etc., as
well as some trace heavy metal ions, which makes it difficult to be
directly used in conventional microalgae cultivation. Microalgae

http://dx.doi.org/10.1016/j.biortech.2016.03.007
0960-8524/Ó 2016 Elsevier Ltd. All rights reserved.
352 C.-D. Zhang et al. / Bioresource Technology 209 (2016) 351–359
Nomenclature
vol. abbreviation of volume
min abbreviation of minute
Ls liter of sample, L
m weight, g
M molecular weight, g mol1
n number of experimental point
p average productivity, g L1 day1
t the time of cultivation
V the total volume of the culture medium, L
VAR.P Excel VAR.P function in Eq. (6)
ct the biomass concentration, mol/L
cCO2 the concentration of carbon dioxide, mol L1
had been screened, isolated and cultivated in order to obtain algae
strains tolerating high CO2 concentration, as well as acid-and
thermo-tolerance (Kurano et al., 1995; Li and Wang, 2015; Morais
and Costa, 2007b; Sakai et al., 1995). It might be a tremendous work
to screen a kind of suitable microalgae from thousands of microal-
gae strains. On the other hand, the question of how to improve the
fixation efficiency of carbon dioxide, especially in open raceway
ponds (abbreviated as ORWPs) breeding manner, should be consid-
ered (Cheng et al., 2013; Li and Luo, 2013). In fact, the open-culture
systems cannot normally make effective use of the supplied CO2,
because of easily escaping from the culture broth due to the limited
depth of liquid layer, short retention time and limited interfacial
area of contact. And the maximum fixation efficiency of CO2 ranged
from 10% to 30% (Becker, 1994; Weissman and Goebel, 1985). An
airlifted-driven raceway reactor was designed by Ketheesan and
Nirmalakhandan (2012) for microalgae cultivation with the maxi-
mum CO2 fixation efficiency of 33%. A carbonation column in con-
junction with open ponds was designed by Putt et al. (2011) to
investigate the absorption efficiency of CO2. A closed multi-stage
sequential photo-bioreactor also improved CO2 fixation efficiency
to 85.6% under the concentration of 15% (vol.) CO2 by Cheng et al.
(2013). A closed raceway pond was rebuilt from a normal open
raceway pond covered with specially designed transparent cover-
plates by Li and Luo (2013). The CO2 fixation efficiency increased
to 95% under the condition of intermittent gas sparging.
According to the discipline of interphase mass transfer (Seader
and Ernest, 1998), the overall mass transfer resistance of CO2 trans-
port is mainly controlled by resistance from the liquid film. It takes
measures to decrease the diameter of liquid drops and thickness of
the liquid films, and increase turbulence of liquid phase. All of
these measures are beneficial for increasing the overall mass trans-
fer coefficient (Carvalho et al., 2006). Based on the Henry’s Law and
theory of electrolyte solution, the higher the CO2 partial pressure
is, the more CO2 is absorbed, thus more carbonate ions is produced
with the lower pH (Luo et al., 2015; Kurano et al., 1995). For the
reasons of the microalgae strains’ tolerance, low CO2 fixation effi-
ciency and high operation cost, the methods mentioned in litera-
ture were unachievable to directly scale up by using the exhaust
emission as the source of supply CO2. In this work, a new-type
CO2 supply method and apparatus was presented to achieve
microalgae cultivation on large scale with high concentration from
20% (vol.) to pure CO2.
2. Materials and experimental method
2.1. Materials
The strain Chlorella pyrenoidosa (FACHB 9) was used in this
work, which was obtained from Wu Han Institute of Hydrobiology,
x experimental value

x mean value of experiments in Eq. (7)
Subscripts
i the i experimental point
inlet the inlet of the spraying tower
outlet the outlet of the spraying tower
Greek alphabet
b CO2 absorption efficiency in Eq. (4)
d root mean square error
g CO2 fixation efficiency in Eq. (3)
an embranchment of Chinese Academy of Science. 8 m3 of 15 cm
height of tap water which contained some inorganic salts was used
as the outdoor cultivation medium. The water medium was circu-
lated along with the two-channel open raceway ponds (abbrevi-
ated as ORWP) by paddlewheels under natural conditions. The
size of the channel raceway ponds is 16.1 m long, 1.6 m wide
and 0.53 m deep. The initial biomass concentration ranged from
0.2 g L1 to 0.3 g L1.
2.2. Experimental apparatus and operation method
A spraying tower with 1.8 m height, 0.8 m diameter and two
spray nozzles on the top was shown in Fig. 1(a). The tower was
used to absorb CO2, and combined with ORWP (1) to provide a car-
bon source for microalgae photoautotrophic cultivation on large-
scale. The main part of the tower is self-made in poly-methyl acry-
late (PMMA), a kind of visual material. The whole tower with its
accessories including pumps, switch box, valves and pipelines,
et al. was amounted on a removable steel stainless platform. The
platform was easily operated, provided that it was powered on
and well connected with inlet and outlet pipes. A running state
of the platform device was shown in Fig. 1(b).
During the operation process, pure CO2 (purity P99%) flowed
from a cylinder (16) and entered into the bottom of the spraying
tower through a T-shaped gas distributor (10). A small proportion
of culture medium with a liquid flow rate about 0.51 m3 h1 (only
account for 6.38% of the total cultivation medium) was suctioned
by diaphragm pumps (3) (model Y2-80M2-4, made by Shanghai
Zhongqiu Pump and Valve Co., Ltd., China) and nebulized by
atomizing nozzles (7) on the top of the spraying tower. The
atomization droplets were in continuous counter flow with the
up current of CO2, which increased the interfacial areas of gas–
liquid phase and the turbulence of the liquid phase. The absorbed
CO2 immediately flowed back via outlet pipe (11) and mixed with
the main microalgae culture medium of ORWP to achieve CO2
complementary. Paddle wheel (2) was used for mixing and stir-
ring the culture medium in the ORWPs. A pressure gauge (4)
was connected with the outlet pipe to determine the liquid pres-
sure of the diaphragm pump. The flow rate of culture medium
(L min1) could be adjusted by a globe value (5) and recorded
by a cumulative flow meter (6) (made in Suzhou Water Meter
Co., Ltd., China). The flow rate (L min1) of CO2 was determined
by a rotameter (14) (Model LZB-6WB, Changzhou Double Thermal
Instrument and Meter Co., Ltd., China). Values of pH of the
microalgae culture medium located at the position of the inlet
and outlet pipe of the spraying tower were measured by an
online pH-meter (Model pH-1186, Shanghai Lianxiang Environ-
mental Protection Technology Co., Ltd., China). The electricity
consumption was measured by an energy meter (Model
 C.-D. Zhang et al. / Bioresource Technology 209 (2016) 351–359 353

Fig. 1a. Flow chart of a spraying tower used for CO2 supplementary in microalgae cultivation.

the spraying absorption tower were recorded every of 2 h during

the experiment period.

2.3. Contrast experiment

The method of evaluating this new apparatus is to cultivate

microalgae in ORWPs via two different CO2 supply ways,
which is conventional process (bubbling, denoted as ORWP-A
mode) and new process (spraying absorption, denoted as ORWP-B
mode).
In ORWP-A mode, carbon source was continuously aerated into
the microalgae medium through sintered porous pipes placed at
the bottom of the pond with concentration of 2–10% vol. CO2-air
mixture to control the pH value in an appropriate range (6.5–
8.5). In ORWP-B mode, the flow chart of which is shown in Fig. 1
(a) described above. Pure CO2 was used because of the short
gas–liquid contact time (within tens of second). The biomass
concentration, productivity, pH values, CO2 flow rate (CO2
consumption) and CO2 fixation efficiency were investigated for
the new spraying process, ORWP-B mode and compared with
that of conventional bubbling process, ORWP-A mode. The experi-
ments were undertaken during the time of September 2014 to
November 2014 under the natural conditions of the air tempera-
ture, the temperature of microalgae medium, light intensity, the
atmosphere pressure and natural wind speed, etc. The average
light intensity varied from 6 Klux to 93.3 Klux and the liquid
medium temperature changed from 16.7 °C to 28.9 °C. And the
other weather data of Jiaxing City in Zhejiang province during
the experimental time in year 2014 can be consulted from the
internet reported history data.

2.4. Analysis methods

Fig. 1b. Photograph of a running state of the spraying absorption tower.

2.4.1. Biomass concentration and average productivity

DTS901, Shanghai People Enterprise (Group) Co., Ltd., China). Dur- Optical density measurement and difference methods (Lee and
ing the period of experimental day time, the inlet and outlet pH Shen, 2004) are two main methods for determining the microalgae
values, inlet flow rate of culture medium and CO2 flow rate for yield. In this work, the difference method was adopted.
354 C.-D. Zhang et al. / Bioresource Technology 209 (2016) 351–359
The biomass concentration is determined by weight method
and calculated by Eq. (1).
m2  m1
Ct ¼
Ls
The average productivity is calculated from the variation in bio-
mass concentration by Eq. (2) (developed by Ketheesan and
Nirmalakhandan, 2012).
Ct  C0
p¼
t  t0
In Eqs. (1) and (2), C0 is initial biomass concentration at time t0
(seeding biomass concentration); Ct is referred to biomass concen-
tration at time t (17:30 each day), g L1; m1 is the weight of empty
weighing bottle, g; m2 is the weight of weighting bottle with
microalgae, g; Ls is the volume of sampling, L; p is the average pro-
ductivity of microalgae, g L1 day1.
2.4.2. CO2 fixation efficiency of microalgae
In order to assess the effectiveness of the CO2 supplying system,
CO2 fixation efficiency determined from the CO2 consumption rate
by algal culture is more appropriate than that from the actual CO2
supplying rate (Ketheesan and Nirmalakhandan, 2012). In this
work, the fixation efficiency is calculated by Eq. (3) (Morais and
Costa, 2007a).
p  mcbm  V  ðM CO2 =M 1
C Þ
g¼  100%
mCO2
where g is the efficiency of CO2 fixation by microalgae, p is the aver-
age productivity of microalgae each day, g L1 day1, mcbm is the
fraction of carbon in the biomass and equates to 0.5 g g1 (Li and
Luo, 2013); V is the total volume of the culture medium, 8000 L;
MCO2 and MC are molar mass of carbon dioxide and carbon, respec-
tively; and mCO2 is the total mass flow of CO2 supply each day,
g day1.
2.4.3. CO2 absorption efficiency of the spraying absorption tower
The efficiency of CO2 fixation by microalgae is highly relative to
CO2 absorption efficiency of the spraying absorption tower.
Besides, absorption efficiency of CO2 is an important parameter
in the design and operation of the spraying tower. The absorption
efficiency b is calculated by Eq. (4).
DcCO2  Lc  V m
b¼  100%
V in  V out
where DcCO2 is the concentration difference of CO2 of inlet and out-
let of the spraying absorption tower (so-called the content of CO2
absorbed), mol L1, and calculated by Eq. (5); Lc is the volume flow
rate of the culture medium injected into the spraying absorption
tower, L min1; Vin and Vout are inlet and outlet flow rates of CO2
into the spraying tower, respectively, L min1; Vm is the molar vol-
ume of CO2 at experimental temperature.
The concentration of CO2 in the liquid culture medium (includ-
ing different form of CO2 in solution) was quantified by measuring
chemical deposition with barium chloride and sodium hydroxide.
Then, the concentration of CO2 absorbed is calculated by
moutlet;CO2  minlet;CO2
DcCO2 ¼
M BaCO3  Ls
where moutlet;CO2 and minlet;CO2 are CO2 content in outlet and inlet of
spraying absorption tower, g; Ls is sample volume, L; M BaCO3 is molar
mass of barium carbonate, 197.34 g mol1.
3. Results and discussion
Cultivation of two batches for C. pyrenoidosa with the ORWP-A
mode and ORWP-B mode was undertaken to compare the new
method with conventional method under the same experimental
conditions. The pH value change, flow rate of CO2 consumption,
biomass concentration and productivity of microalgae, efficiency
of CO2 fixation are used as the criteria to assess the two CO2 sup-
ð1Þ
plying facilities. Additionally, absorption efficiency of CO2 and
energy consumption of the spraying absorption tower were also
evaluated, which indicated that this is an actual feasible scheme
of microalgae cultivation on large-scale or not.
ð2Þ 3.1. Biomass concentration and productivity of microalgae, pH and
CO2 flow rate
The relationship of biomass concentration and productivity
with cultivation days for the two batches microalgae breeding
were shown in Fig. 2(a) and (b).
As shown in Fig. 2(a-1) and (b-1), the biomass concentrations of
microalgae cultivation in ORWP-B mode were almost greater than
that of ORWP-A mode for the two experimental batches. Under the
same identical experiment conditions, the maximum biomass con-
centration was 0.927 g L1 (the 8th day) for the first batch, and
0.4809 g L1 (the 5th day) for the second batch with ORWP-B
mode; but only 0.7867 g L1 (the 7th day) and 0.3545 g L1 (the
4th day) were obtained for the corresponding two batches with
ORWP-A mode. The variation trend of the biomass concentration
increased with cultivation days from the beginning to the 8th
day for the first batch, and then decreased after this day. From
ð3Þ the view point of microalgae breeding, the microalgae should be
harvested in this day.
The calculated average productivity of microalgae with ORWP-
A mode and ORWP-B mode were shown in Fig. 2(a-2) and (b-2),
respectively. Unlike the variations of biomass concentration curve,
the average productivity increased relatively large at the first two
days, maintained nearly unchanged till to the maximum value; and
then decreased afterwards. The maximum productivity appeared
at values of 0.1144 g L1 day1 and 0.0622 g L1 day1 for the
two batches with ORWP-B mode; and only of 0.081 g L1 day1
and 0.0581 g L1 day1 with ORWP-A mode. The different results
of the two batches were due to the different growing circumstance.
During the first batch of microalgae cultivation days, it was always
in warm sunshine. But for the second batch, it was sometimes in
chilly rainy days.
Ordinarily, the shear action of nozzles may be strongly harmful
to microalgae. As is seen from of Fig. 3, the result of C. pyrenoidosa
ð4Þ
cultivation with ORWP-B mode is better than that of ORWP-A
mode. That is to say, the CO2 supply system with a spraying tower
is suitable for C. pyrenoidosa cultivation.
The compared biomass concentration and productivity of C.
pyrenoidosa cultivation in this work with the results reported in lit-
eratures is listed in Table 1. Under the condition of (6–12)% (vol.)
CO2 as a carbon complementary, the maximum biomass concen-
tration of (0.98–1.14)g L1 for Chlorella Kessleri in a 1.8 L conical
flash photo-bioreactor (PBR) was obtained (Morais and Costa,
2007b). A maximum productivity of 0.515 g L1 day1 for Chlorella
sp. MTF-15 was achieved in a cylindrical acrylic polymer column of
1 L with (23–27)% (vol.) CO2 as a carbon source (Kao et al., 2014).
The maximum biomass concentration and average productivity
ð5Þ of 0.927 g L1 and 0.114 g L1 day1 for C. pyrenoidosa were gained
with ORWP-B mode under the condition of pure CO2 as a carbon
complementary, in which are lower than those listed in Table 1.
However, results reported by the International Energy Agency
(2011) showed that most high levels of sustained productivity
for microalgae cultivation in an open pond were in the range of
15–40 g m2 day1 (already converted to be 0.05–0.133 g L1
day1 in 0.3 m depth of ORWPs). Cultivation of microalgae on large
scale is mostly constrained by the low ratio of contacting area of
gas–liquid two-phase to volume of RWP. Therefore, the productiv-
ity of per unit volume is also reduced (Chiaramonti et al., 2013).
 C.-D. Zhang et al. / Bioresource Technology 209 (2016) 351–359 355

Fig. 2. The relationship of biomass concentration and productivity with cultivation days for two batches microalgae cultivation with ORWP-A and ORWP-B modes. (a) The
first batch of microalgae cultivation; (b) the second batch of microalgae cultivation.

Based on the viewpoint of scale-up effect and high CO2 concentra-
tion as the carbon complementary, the biomass concentration and
productivity in this work is acceptable. Compared with the results
obtained by Cheng et al. (2015), the maximum productivity of Nan-
nochloropsis oculata cultivation was 0.097 g L1 day1 by using real
flue gas as the carbon complementary.
Unlike the previous study concerning CO2-supply (Putt et al.,
2011), the pH value of microalgae broth could be controlled within
a certain range. While, it is difficult to control the pH steadily at a
certain value because the cultivation conditions, such as light
intensity, temperature, atmosphere pressure, and natural wind
speed, etc can’t be artificially controlled outdoors in this work.
Variations of pH value with cultivation days for C. pyrenoidosa by
ORWP-A mode and ORWP-B mode were shown in Fig. 3
(a) and (b). The variations of pH value for both ORWP-A mode
and ORWP-B mode were all in range from 6.5 to 8.5, which is suit-
able for the microalgae growth. The variance analysis for pH value
by VAR.P function with Microsoft Excel software was calculated
by:
ðx1 
xÞ2 þ ðx2 
xÞ2 þ ::: þ ðxn 
xÞ2
VAR:Pðx1 ; x2 ; :::; xn Þ ¼ ð6Þ
n
356 C.-D. Zhang et al. / Bioresource Technology 209 (2016) 351–359

tion d is defined as the square root of VAR.P function. The results

of variance analysis for pH value shown in Fig. 3(a) and (b) were
listed in Table 2. The calculated value of VAR.P function for the
two batches cultivation with ORWP-B mode was both 0.06, which
was less than 0.10 and 0.07 with ORWP-A mode for the correspond-
ing two batches. This meant that the cultivation of microalgae with
ORWP-B mode is more suitable on large scale under the condition of
the growing circumstance with a relative stable pH value.
Relationship of average CO2 flow rates with cultivation days for
ORWP-A mode and ORWP-B mode was shown in Fig. 4(a) and (b).
As is seen from Fig. 4(a) and (b), the average CO2 flow rate used in
the ORWP-B mode was 2.62 L min1 for the first batch, and
2.63 L min1 for the second batch; while 9.39 L min1 and
7.0 L min1 (counted in pure CO2) of CO2 flow rate were requested
in ORWP-A mode for the corresponding two batch cultivation,
respectively. Therefore, under the same identical experiment con-
ditions, the average CO2 flow rate of ORWP-B mode was less than
nearly three times of ORWP-A mode. The results of variance anal-
ysis for CO2 flow rate were listed in Table 3. The calculated values
of VAR.P function for the first and the second batch cultivation
with ORWP-B mode were 0.24 and 0.38, which were all less than
6.35 and 10.15 with ORWP-A cultivation mode for the correspond-
ing two batches. Additionally, the steady CO2 gas supply is benefi-
cial for operated with automatic control instrument on large-scale
cultivation of microalgae with ORWP-B mode.

3.2. Average efficiency of CO2 fixation by microalgae

Comparisons of the calculated average efficiency of CO2 fixation

in this work with the result estimated from literatures was listed in
Table 4. It can be seen from Table 4, the average fixation efficiency
of CO2 for ORWP-B mode was 50.0%, which was obviously higher
than 11.17% for ORWP-A mode. The influence of factors on CO2 fix-
ation efficiency is very complicated, which includes CO2 supplying
system, CO2 concentration, microalgae strains, photo bioreactor
(PBR) and uncontrolled natural conditions (light, temperature)
and so on.
By using a closed raceway pond with a transparent cover and
intermittent gas sparging, the maximum CO2 fixation of Chlorella
vulgaris cultivation might be 95% (Li and Luo, 2013). The highest
CO2 fixation efficiency can be 85.6% by using a multi-stage sequen-
tial bioreactor for Chlorella PY-ZUI cultivation (Cheng et al., 2013).
Fig. 3. Variations of pH value with cultivation days for Chlorella pyrenoidosa by
ORWP-A and ORWP-B modes. (a) The first batch of microalgae cultivation; (b) the
Two main methods of increasing CO2 fixation efficiency are reduc-
second batch of microalgae cultivation. tion of the gas bubble diameter size (Kao et al., 2014) and increase
of the retention time (Morais and Costa, 2007a). However, the
x1 þ x2 þ ::: þ xn method of increasing CO2 concentration was restricted by the

x ¼ ð7Þ
n acidic endurance capability of microalgae strains. Besides, high
energy consumption is ordinary required to reduce gas bubble
where xi (i = 1, 2, . . ., n) is the experimental value at the point i.

x is diameter. Although the various types of experimental photo-
the mean value of the experiments. The root-mean-square devia- bioreactors (PBRs) studied in Table 4 may have high CO2 fixation

Table 1
Comparison of biomass concentration and productivity in algae cultivation systems: literature reported vs. this work.

References Species of algae Cultivation apparatus Culture medium Vol.% in air ct,max (dry paver.,max/
volume/L base)/g L1 g L1 d1
Ketheesan and Scenedesmus sp. Airlift-raceway 23 1 1.43 0.085
Nirmalakhandan (2012)
Morais and Costa (2007b) Chlorella kessleri Conical flask photo-bioreactor 1.8 6 0.98 0.087
Scenedesmus obliquus Conical flask photo-bioreactor 12 1.14 0.076
Li and Luo (2013) Chlorella vulgaris Closed raceway 12 15 – 0.350
pond/continuous operation
Kao et al. (2014) Chlorella sp. MTF-15 Column-type glass fabricated 1 23–27, flue gas from 2.523 0.515
photobioreactor coke oven
Cheng et al. (2015) Nannochloropsisoculata Open raceway pond 3.1  105 Flue gas – 0.1015
Chiu et al. (2008) Chlorella sp. Cylindrical glass reactor 0.8 2 1.445 0.422
This study (ORWP-B) Chlorella pyrenoidosa ORWP-spray absorption tower 8000 Purity P99.5 0.927 0.114
This study (ORWP A) Chlorella pyrenoidosa ORWP-spray absorption tower 8000 5–10 0.787 0.081
 C.-D. Zhang et al. / Bioresource Technology 209 (2016) 351–359 357

Table 2 Table 3
Variance analysis of pH value for ORWP-A and ORWP-B modes. Variance analysis of CO2 flow rate of ORWP-A and ORWP-B modes.

ORWP Batch (a)a Batch (b)b ORWP Batch (a)a Batch (b)b
mode mode
Max Min Average VAR.P Max Min Average VAR.P Max Min Average VAR.P Max Min Average VAR.P
A 8.46 6.80 7.25 0.10 7.80 6.65 7.14 0.07 A 14.0 3.5 9.39 6.35 15.0 3.0 7.00 10.15
B 7.83 6.88 7.25 0.06 8.09 6.95 7.29 0.06 B 4.0 2.0 2.63 0.24 4.0 2.0 2.62 0.38
a a
Experimental batch (a), the initial concentration of algae: both ORWP-A and Experimental batch (a), the initial concentration of algae: both ORWP-A and
ORWP-B are 0.22 g L1; light intensity: 6–93.3 Klux, average: 41.7 Klux; tempera- ORWP-B are 0.22 g L1; light intensity: 6–93.3 Klux, average: 41.7 Klux; tempera-
ture: 16.7–28.9 °C, average: 23.6 °C; circulation flow of culture medium: 4.31– ture: 16.7–28.9 °C, average: 23.6 °C; circulation flow of culture medium: 4.31–
19.02 L min1, average: 8.22 L min1. 19.02 L min1, average: 8.22 L min1.
b b
Experimental batch (b), the initial concentration of algae: both ORWP-A and Experimental batch (b), the initial concentration of algae: both ORWP-A and
ORWP-B are 0.2 g L1; light intensity: 11.2–89 Klux, average: 48.41 Klux; temper- ORWP-B are 0.2 g L1; light intensity: 11.2–89 Klux, average: 48.41 Klux; temper-
ature: 17.9–28 °C, average: 23.8 °C; circulation flow of culture medium: 5.89– ature: 17.9–28 °C, average: 23.8 °C; circulation flow of culture medium: 5.89–
14.51 L min1, average: 8.95 L min1. 14.51 L min1, average: 8.95 L min1.

3.3. CO2 absorption efficiency of the spraying tower

The relationship between CO2 concentration in the inlet and

outlet algae solution of the spray tower, as well as absorbed CO2
content for the two batches cultivation with time was shown in
Fig. 5(a) and (b). And accordingly, the CO2 absorption efficiency
of the spraying tower calculated by Eq. (4) was depicted in Fig. 6
(a) and (b).
As is seen from Fig. 5(a) and (b), CO2 concentration in the out-
let algae solution (Dcoutlet; CO2 ) of the spraying tower for the two cul-
tivation batches was in the range from 0.018 mol L1 to
0.019 mol L1; while the CO2 concentration in the inlet algae solu-
tion (Dcinlet; CO2 ) of the tower slowly increased with the cultivation
days. Consequently, the curve of the absorbed CO2 concentration
(DcCO2 ) calculated from Eq. (5) decreased gradually with the culti-
vation days.
As shown in Fig. 6(a) and (b), the absorption efficiency of the
spraying tower fluctuated within a definite range, and the average
absorption efficiency of the spraying tower was 77.50% and 73.71%
for the two batches, respectively. The CO2 absorption efficiency is
closely connected with the quantity of circulating algae solution.
Larger quantity of circulating algae solution might lead to form
bigger liquid droplets and lessen gas–liquid contacting areas. As
a result, the CO2 absorption efficiency decreases rapidly. While,
in this work, under the conditions of the circulating algae solution
flow rate ranged from 4.31 L min1 to 19.02 L min1, there was no
significant effect on the CO2 absorption efficiency.
Ordinarily, there is a phenomenon that pH of culture medium
increases gradually with microalgae growing. The reason might
be that microalgae growth consumes a great amount of anions in
culture medium, such as NO  2
3 and H2PO4 , HPO4 to form biomass.
Thus, the dissolved CO2 must convert to its anions form, such as
HCO 2
3 and CO3 , to keep ionic balance with cations (like Na ,
+

et al.). The culture medium will be alkalized with too much anionic
CO23 . An excessive amount of CO2 must be introduced into the
spraying tower to control the pH value. Otherwise, algal cells are
much more likely injured in a high pH culture medium.

Fig. 4. Relationship of average CO2 flow rates with cultivation days for both ORWP-
A and ORWP-B modes. (a) The first batch of microalgae cultivation; (b) the second
batch of microalgae cultivation.
efficiency, it is actually very difficult to enlarge on industrial scale.
In this work, a spraying absorption tower designed for C. pyrenoi-
dosa cultivation on large-scale provided a different way with high
CO2 fixation efficiency, where a little microalgae culture broth
required is dispersed in pure CO2 gas supply flow at a very short
time.
3.4. Energy consumption of the spraying absorption tower
Capital cost and operating charges are the two indexes to eval-
uate the microalgae cultivation process feasibility and economics
(Chiaramonti et al., 2013). Captial cost refers to the all equipment
value. And the operating charges main include the cost of CO2 and
electricity consumption. The consumption of CO2 was compared
above for the two cultivation modes. And in this part, power con-
sumed by the two diaphragm pumps for transporting a proportion
of culture medium into the spraying absorption tower was only
evaluated. Based on the original record of experiment data, the
358 C.-D. Zhang et al. / Bioresource Technology 209 (2016) 351–359

Table 4
Comparison of CO2 fixation efficiencies in algae cultivation systems: literature reported vs. this work.

References Species of algae Cultivation apparatus Culture medium Vol.% in air gmax (%) gaver: (%)
volume /L
Ketheesan and Nirmalakhandan (2012) Scenedesmus sp. Airlift-raceway 23 0.25 33 20
Morais and Costa (2007a) Spirulina sp. Three-stage serial tubular photo-bioreactor 1.8  3 6 53.29 37.9
Scenedesmus obliquus 28.08 13.45
Li and Luo (2013) Chlorella vulgaris Closed raceway pond/intermittent operation 12 15 95 –
Chiu et al. (2008) Chlorella sp. Cylindrical glass reactor 0.8 2 58 –
Cheng et al. (2013) Chlorella PY-ZU1 Multi-stage sequential bioreactor 1 15 85.6 –
This work (ORWP-B) Chlorella pyrenoidosa ORWP-spray absorption tower 8000 100 73.11 50
This work (ORWP-A) Chlorella pyrenoidosa ORWP with bubbling 8000 5–10 15.13 11.17

Fig. 5. The relationship between CO2 concentration in the inlet and outlet algae
solution of the spray tower as well as absorbed CO2 content with cultivation days.
(a) The first batch of microalgae cultivation; (b) the second batch of microalgae
cultivation.
electricity consumption was about 2.1 kWh per day on average for
the first batch of microalgae cultivation; and the maximum elec-
tricity consumption was nearly 3.19 kWh per day. The evaluated
energy consumption for the two batches microalgae cultivation
Fig. 6. The calculated absorption efficiency of CO2 in the spraying tower for two
batches microalgae cultivation with ORWP-B mode. (a) The first batch of microalgae
cultivation; (b) the second batch of microalga cultivation.
process was listed in Table 5. Depending on the latest National
industrial electricity price of Zhejiang province, 0.946 RMB per
kWh, the total electricity bill was only 1.987 RMB per day. Com-
pared with microalgae cultivation with ORWP-A mode, this new
apparatus may consume less energy. Because the ORWP-A mode
usually needs CO2-air mixture gas as the CO2 supplementary, and
air flow is generated from an air-compressor working all days.
 C.-D. Zhang et al. / Bioresource Technology 209 (2016) 351–359 359

Table 5 Chiaramonti, D., Prussi, M., Casini, D., Tredici, M.R., Rodolfi, L., Bassi, N., Zittelli, G.C.,
Electricity consumption estimated for the spray absorption tower algae cultivation. Bondioli, P., 2013. Review of energy balance in raceway ponds for microalgae
cultivation: re-thinking a traditional system is possible. Appl. Energy 102, 101–
Batch Electricity consumption/kW h1 111.
Chiu, S.Y., Kao, C.Y., Chen, C.H., Kuan, T.C., Ong, S.C., Lin, C.S., 2008. Reduction of CO2
Max Min Average Total amount
by a high-density culture of Chlorella sp. in a semi continuous photo bioreactor.
ORWP-B(a) 3.19 1.42 2.07 18.68 Bioresour. Technol. 99, 3389–3396.
ORWP-B(b) 2.58 1.84 2.25 11.25 IEA Bioenergy Task 39 and IEA AMF, 2011. Algae as a Feedstock for Biofuels – An
Assessment of the Current Status and Potential for Algal Biofuels Production
(www.task39.org, access 03.12.11).
Kao, C.Y., Chen, T.Y., Chang, Y.B., Chiu, T.W., Lin, H.Y., Chen, C.D., Chang, J.S., Lin, C.S.,
4. Conclusions 2014. Fixation of carbon dioxide in industrial flue gases for the cultivation of
microalga Chlorella sp. Bioresour. Technol. 166, 485–493.
Keffer, J.E., Kleinheinz, G.T., 2002. Use of Algae vulgaris for CO2 mitigation in
A new CO2 supply system of a spraying absorption tower com- autophotobioreactor. J. Ind. Microbiol. Biotechnol. 29 (5), 275–280.
bined with an outdoor ORWP for microalgae photoautotrophic cul- Ketheesan, B., Nirmalakhandan, N., 2012. Feasibility of microalgae cultivation in a
pilot-scale airlift-driven raceway reactor. Bioresour. Technol. 108, 196–202.
tivation is developed in this paper. The microalgae yield, Kurano, N., Ikemoto, H., Miyashita, H., Hasegawa, T., Hata, H., Miyachi, S., 1995.
productivity and CO2 fixation efficiency were investigated, and Fixation and fixation of carbon dioxide by microalgal photosynthesis. Energy
compared with bubbling method. The maximum yield and produc- Convers. Manage. 36 (6/9), 689–692.
Lee, Y.K., Shen, H., 2004. Basic culturing techniques. In: Richmond, A. (Ed.),
tivity of biomass were achieved 0.927 g L1 and 0.114 g L1 day1,
Handbook of Microalgal Culture Biotechnology and Applied Phycology.
respectively. The fixation efficiency of CO2 by microalgae with the Blackwell Publishing, Iowa, p. 43.
spraying tower reached 50%, whereas only 11.17% for bubbling Li, D.J., Wang, L., 2015. Improving high carbon dioxide tolerance and carbon dioxide
fixation capability of Algae sp. by adaptive laboratory evolution. Bioresour.
method. Pure CO2 can be used in the spraying tower. It shows that
Technol. 185, 269–275.
this method of quantifiable control CO2 supply can meet the Li, S.W., Luo, S.J., 2013. Efficiency of CO2 fixation by microalgae in a closed pond.
requirements of microalgae cultivation on large-scale. Bioresour. Technol. 136, 267–272.
Luo, M.Y., Yang, J.H., Gong, Q.T., 2015. Experimental study on CO2 fixation from
simulated flue gas by algae vulgaris. Chem. Ind. Eng. Process. 34 (4), 1147–1151.
Acknowledgements Morais, M.G.D., Costa, J.A.V., 2007a. Biofixation of carbon dioxide by Spirulina sp.
and Scenedesmus obliquus cultivated in a three-stage serial tubular
This research was funded by National Basic Research Program photobioreactor. J. Biotechnol. 129 (3), 439–445.
Morais, M.G.D., Costa, J.A.V., 2007b. Isolation and selection of microalgae from coal
China (973 Program: 2011CB200904), National High Technology fired thermoelectric power plant for biofixation of carbon dioxide. Energy
Research and Development Program (863 Programs: Convers. Manage. 48, 2169–2173.
2012AA050101), and supported by Jiaxing Zeyuan Bio-Products Putt, R., Singh, M., Chinnasamy, S., Das, K.C., 2011. An efficient system for
carbonation of high-rate algae pond water to enhance CO2 mass transfer.
Limited Company. Bioresour. Technol. 102, 3240–3245.
Sakai, N., Sakamoto, Y., Kishimoto, N., Chihara, M., Karube, I., 1995. Chlorella strains
References from hot springs tolerant to high temperature and high CO2. Energy Convers.
Manage. 36 (6–9), 693–696.
Seader, J.D., Henley, Ernest J., 1998. Separation Process Principles. John Wiley & Sons
Becker, E.W., 1994. Microalgae: Biotechnology and Microbiology. Cambridge
Inc, New York.
University Press.
The industrial electricity average price in Zhejiang province, 2014. http://
Carvalho, A.P., Meireles, L.A., Malcata, F.X., 2006. Microalga reactors: a review of
www.dongtou.gov.cn/zxfw/qyfw/ggsy/sdq/20140613/338858_1.htm (access:
enclosed systems designs and performances. Biotechnol. Progr. 22, 1490–1506.
06.13.2014).
Chae, S.R., Hwang, E.J., Shin, H.S., 2005. Single cell protein production of Euglena
The history weather in Jiaxing, Zhejiang province, 2014. http://www.tianqihoubao.
gracilis and carbon dioxide fixation in an innovative photo-bioreactor.
com/lishi/jiaxing/month/201410.html.
Bioresour. Technol. 97 (2), 322–329.
Watanabe, Y., Ohmura, N., 1992. Isolation and determination of cultural charac-
Cheng, J., Huang, Y., Feng, J., 2013. Improving CO2 fixation efficiency by optimizing
teristics of microalgae which functions under CO2 enriched atmosphere. Energy
algae PY-ZU1 culture conditions in sequential bioreactors. Bioresour. Technol.
Convers. Manage. 33 (6–8), 545–552.
144, 321–327.
Weissman, J.C., Goebel, R.P., 1985. Production of Liquid Fuels and Chemicals by
Cheng, J., Yang, Z.B., Huang, Y., Huang, L., Hu, L.Z., Xu, D.H., Zhou, J.H., Chen, K., 2015.
Microalgae. Solar Energy Research Institute, Golden Co (SERI/STR-231-2649).
Improving growth rate of microalgae in a 1191 m2 raceway pond to fix CO2
from flue gas in a coal-fired power plant. Bioresour. Technol. 190, 235–241.