Journal of Bioscience and Bioengineering

VOL. xxx No. xxx, xxx, xxxx

www.elsevier.com/locate/jbiosc

Outdoor cultivation of Chlorella sp. in an improved thin-film flat-plate

photobioreactor in desertification areas

Chenghu Yan, Zhihui Wang, Xia Wu, Shumei Wen, Jing Yu, and Wei Cong*

State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China

Received 29 October 2019; accepted 8 December 2019

Available online xxx
In order to reduce the flow resistance of the thin-film flat plat photobioreactor (FPPBR) and make it more suitable for
mass microalgae cultivation, the channel diameter was modified to 0.06 m and the thin-film FPPBR consisted of 10
parallel shunt-wound channels. A thin-film FPPBR system with 100 modified FPPBRs was constructed and used for
Chlorella sp. cultivation in desertification areas (Ordos, China) from July to September of 2018. The pressure drop of the
modified FPPBR system decreased significantly and the microalgae showed much higher productivity. The pressure drop
was about 11.8 kPa when the liquid velocity was 0.238 m sL1. The final biomass concentration and area productivity
reached 2.01 g LL1 and 49.79 g mL2 dayL1 respectively, and the yearly productivity of Chlorella sp. was estimated to be
about 15.24 t haL2 yearL1. The results demonstrated that high productivity of Chlorella sp. could be achieved in the
improved FPPBR system in desertification areas and the improved FPPBR system was feasible for mass cultivation of
microalgae in the commercial application.
Ó 2019, The Society for Biotechnology, Japan. All rights reserved.

[Key words: Outdoor cultivation; Flat plate photobioreactor; Mass production; Chlorella sp.; Baffle]

Microalgae are photosynthetic microorganisms from either
marine or fresh water environments, which have been used as
feedstock for high value compounds and biofuels production since
they are able to transform carbon dioxide into cellular biomolecules
such as biofuels, antioxidant compounds, nutraceuticals, animal/
fish feeds, and fertilizers via photosynthesis (1e4). Thus, the de-
mand for algae biomass is increasing in recent years. For mass
production of microalgae, efficient photobioreactors with low en-
ergy input and utilizing maximal solar radiation are required (5,6).
Up to now, a large number of photobioreactors with different vol-
umes and shapes have been developed (7e10). However, only a few
of them have been used for commercial microalgae cultivation.
In our previous study, a novel thin-film FPPBR mounted with
inclined baffles was developed and used for Chlorella sp. cultivation
(11,12). The biomass productivity of Chlorella sp. in the novel FPPBR
was 25.2% higher than the FPPBR without baffles, and the biomass
productivity of Chlorella sp. and Scenedesmus dimorphus in the
photobioreactor system reached 11 g m
2 day
1 and 21.9 g m
2
improved by increasing the diameter of each shunt-wound
channel (13). Moreover, culture efficiency is also related to
channel diameter, for maximum production in a given area, the
suggested channel diameter is about 0.06 m (14,15). Therefore, in
order to make the thin-film flat plat photobioreactor (FPPBR)
more suitable for commercial microalgae cultivation, the channel
diameter was modified to 0.06 m and the thin-film FPPBR con-
sisted of 10 parallel shunt-wound channels. An improved thin-
film FPPBR system with 100 FPPBRs was built up and used for
Chlorella sp. cultivation at Engebei (Ordos, China), close to the
Kubuqi Desert. The microalgae cultivation experiments were
performed for three months (July to September 2018), and the
relationships between biomass productivities and environmental
conditions were investigated.
MATERIALS AND METHODS

day
1. However, the flow resistance of microalgae suspension in the

photobioreactor is still high, which increases the pressure of the
photobioreactor and the risk of being damaged, thus the phobior-
eactor system can only be operated in low flow velocity, resulting in
the decrease of flashing light effect and productivity. Thus, the
photobioreactor system remain needs to be improved.
The configuration of the thin-film FPPBR is similar to Z-type
flat plate manifolds, which had demonstrated that the flow rate
uniformity and the pressure drop of Z-type manifolds could be
* Corresponding author. Tel.: þ86 10 82627060; fax: þ86 10 82627066.
E-mail address: weicong@ipe.ac.cn (W. Cong).
Configuration of the thin-film FPPBR and photobioreactor system In
order to reduce the flow resistance of the photobioreactor and the pressure drop of
the photobioreactor system, the thin-film FPPBR reported in our previous work was
modified (12), and a photobioreactor system was set up based on the modified
FPPBR, which are shown in Figs. 1 and 2. The unmodified thin-film FPPBR is
shown in Fig. 1C, which is 2.0 m length and 0.96 m width, consisting of sixteen
shunt-wound channels, the length of each channel is 1.44 m. In each channel, the
inclined baffles are attached to both films, which are perpendicular to the film
surface (12). The modified thin-film FPPBR is shown in Fig. 1A and B, which is
2.0 m length and 1.0 m width, consisting of 10 parallel shunt-wound channels.
The baffles are mounted with an inclined angle a ¼ 30 shown in Fig. 1, and the
dimension of the inclined baffles is length (l) ¼ 60 mm, height (h) ¼ 8 mm, width
(w) ¼ 2.5 mm.
In the modified photobioreactor system, an axial-flow pump is used to drive
microalgae suspension, an earth pond of 10 m3 is built as medium storage tank, the

1389-1723/$ e see front matter Ó 2019, The Society for Biotechnology, Japan. All rights reserved.
https://doi.org/10.1016/j.jbiosc.2019.12.007

Please cite this article as: Yan, C et al., Outdoor cultivation of Chlorella sp. in an improved thin-film flat-plate photobioreactor in desertification
areas, J. Biosci. Bioeng., https://doi.org/10.1016/j.jbiosc.2019.12.007
2 YAN ET AL. J. BIOSCI. BIOENG.,

FIG. 1. Schematic diagram and photographs of the thin-film flat plate photobioreactor (FPPBR). (A) Schematic diagram of the improved photobioreactor: 1, inlet; 2, outlet; 3, nether
baffle; 4, upper baffle; 5, flow channel partition; 6, flow channel; 7, inlet cavity; 8, outlet cavity. (B) Improved photobioreactor; (C) photograph of unmodified photobioreactor.

pond is then lined with a 0.15 mm thick polyethylene films, and a polyethylene film
is used to cover the pond in order to counter adverse weather conditions such as
raining and freezing temperature. A gas sparger is installed at the inlet of the first
photobioreactor, pure CO2 is injected into the photobioreactor directly during
microalgae cultivation. Dissolved oxygen and the temperature are controlled using
similar methods reported in our previous research (12). Dissolved oxygen was
regulated by changing the rate of air flow entering the medium tank using suitable
valves and flowmeter through a gas sparger located at the bottom of the medium
tank. A temperature control system with an electromagnetic valve was used to keep
the maximum culture temperature lower than 35  C, When the temperature of the

FIG. 2. Schematic diagram (A) and photographs (B, C) of the pilot scale thin-film flat plate photobioreactor (FPPBR) system. Components: 1, medium storage tank; 2, axial pump; 3,
photosynthetic reaction unit; 4, gas sparger (for O2 removal); 5, gas sparger (for CO2 supply); 6, temperature control system.

Please cite this article as: Yan, C et al., Outdoor cultivation of Chlorella sp. in an improved thin-film flat-plate photobioreactor in desertification
areas, J. Biosci. Bioeng., https://doi.org/10.1016/j.jbiosc.2019.12.007
VOL. xxx, xxxx
TABLE 1. The pressure drop between the inlet and outlet of the photobioreactor system with respect to liquid velocity in the channels.
Frequency (Hz) Pump capacity (m3 h
1) Inlet pressure (kPa)
34.90 11.310.27 8.20.16
36.45 11.940.12 9.10.13
38.50 14.360.17 8.80.15
40.10 20.193.07 10.40.31
42.05 21.180.26 10.80.17
44.95 25.021.02 11.80.11
Data are shown as mean  SD, n ¼ 5.
microalgae suspension exceeded 30  C, the electromagnetic valve opened and
cooling was accomplished by sprinkling the surface of the photobioreactor with
natural ground water. The improved photobioreactor system covers an area of about
250 m2, the photosynthetic reaction unit comprises 100 horizontal improved
FPPBRs, which is arranged into 2 rows, as shown in Fig. 2. The working volume is
about 13 m3 and the illuminated surface area is 119 m2.
The pressure drop and liquid velocity of the thin-film FPPBR system The
pressure drop between the inlet and outlet of the improved photobioreactor system
at different liquid velocity in the photobioreactor was measured. The pressure drop
was calculated by measuring the height difference of water column between the
inlet and outlet of the FPPBR system, the average values and standard deviations
were calculated for five measurements. The liquid velocity in the photobioreactor
was controlled using a frequency converter and measured by solid particle tracing
method, the average values and standard deviations of liquid velocity were calcu-
lated based on the measurement of five particles.
Cultivation experiment and analytical methods The improved thin-film
FPPBR system was located at Engebei (Ordos, China), batch culture of Chlorella sp.
was carried out during the summer and autumn months (July to September) of
2018. The microalgae were incubated for 5e8 days, a part of resulted culture was
harvested, and a part of remaining culture was used as the seed culture for next
batch. The strain Chlorella sp. FACHB-1514 was obtained from Institute of
Hydrobiology, Chinese Academy of Sciences. Chlorella sp. was cultivated in BG 11
medium, the nutrients of which had been reported in our previous research (12).
Natural ground water was filtered by PP cotton filter element with bore diameter
of 1 mm, and was treated with sodium hypochlorite (1 mL L
1) and sodium
thiosulphate. The biomass concentration was calculated by optical density at
680 nm (OD680) of microalgae suspension to monitor the microalgae growth of
the photobioreactor system according to our previous work (12), the average
values and standard deviations of biomass concentration were calculated for three
measurements. The proportional coefficients between biomass concentration and
optical density were 0.29 with correlation coefficients of 0.999 for Chlorella sp.
During the cultivation, the OD680 value was measured four times a day at 8:00,
11:00, 15:00 and 19:00, respectively, and the proportional coefficients were
measured every half month. The area productivity of microalgae Y can be
calculated by
Y ¼ ½ðX2
X1 Þ = ðt2
t1 ÞV = A (1)
where X1 and X2 are biomass concentration of microalgae at time t1 and t2,
respectively. V is the working volume and A is the illuminated surface area of the
thin-film FPPBR.
FIG. 3. Batch culture of Chlorella sp. in thin-film flat plate photobioreactor (FPPBR) system. (A) Biomass concentration; (B) area productivity of Chlorella sp. Data are shown as
mean  SD, n ¼ 3.
Please cite this article as: Yan, C et al., Outdoor cultivation of Chlorella sp. in an improved thin-film flat-plate photobioreactor in desertification
areas, J. Biosci. Bioeng., https://doi.org/10.1016/j.jbiosc.2019.12.007
CULTIVATION OF CHLORELLA SP. IN FPPBR 3
Outlet pressure (kPa) Pressure drop (Pa) Velocity (m s
1)
0 1643.2 0.1190.030
0 1822.6 0.1230.029
0 1763.0 0.1500.017
0 2086.2 0.1820.023
0 2163.4 0.2110.026
0 2362.2 0.2380.029
RESULTS AND DISCUSSION
The pressure drop between the inlet and outlet of the
photobioreactor system The pressure drop between the inlet
and outlet of the photobioreactor system with respect to liquid
velocity is plotted in Table 1. Although the pressure drop increased
as liquid velocity increased, the values were low, even at the
highest velocity 0.238 m s
1, the pressure drop was about
11.8 kPa, while that of the photobioreactor system reported in our
previous research was 21 kPa when the liquid velocity was
0.22 m s
1. The results demonstrated that the pressure of the
FPPBR in the improved photobioreactor system was much
decreased, which made it possible to cultivate microalgae at a
high velocity for longer time.
Outdoor cultivation of microalgae in the improved thin-film
FPPBR system To validate the scalability and performance of the
improved thin-film FPPBR system, batch cultures were performed
from July to September of 2018. The results are shown in Fig. 3.
The microalgae showed steady growth, and the final biomass
concentration and area productivity reached 2.01 g L
1 and
49.79 g m
2 day
1. Several new industrial flat panels made in
transparent plastics have been reported, such as the airlift flat
panel system with internal baffles (Subitec Company, Stuttgart,
Germany) (16) and ProviApt photobioreactor (Proviron Industries,
Hemiksem, Belgium) (17) , while their productivities were about
15 g m
2 day
1, and 31.79 g m
2 day
1 for Chlorella vulgaris and
Nannochloropsis oceanica cultivation, respectively. The results
demonstrated that production of Chlorella sp. can be carried out
outdoors and high productivity could be achieved in the
improved thin-film FPPBR. One reason is the optimization of the
channel diameter, which has been reported that when the
channel diameter is 0.06 m, the maximum productivity in a given
area could be achieved (14,15). On the other hand, with the
4 YAN ET AL.
decrease of the pressure drop of the photobioreactor system, the
microalgae suspension flows in a high velocity in the FPPBR, the
higher frequency of the flashing light effect was produced, which
may enable microalgae growth rate (11).
The effect of seasonal variations of environmental conditions
such as temperature and solar radiation on Chlorella sp. cultivation
was investigated. The weather and temperature variation during the
cultivation were obtained from a Weather forecast web (http://
tianqi.2345.com) as shown in Fig. 4. The daily solar radiation was
divided into 5 levels according to the weather forecast, and each
level has different value: sunny as 500 , sunny interval as 4, cloudy as 3,
cloudy to overcast as 2, rainy as 100. The averaged light intensity of the
levels 4 and 5, levels 2 and 3, and level 1were about
1100 mmol m
2 s
1, 550 mmol m
2 s
1, and lower than
220 mol m
2 s
1, respectively. In the same batch, the productivity
was highest when the solar radiation level was higher than level 4,
followed by the levels 3 and 2, while the productivity was near to
0 when the solar radiation level was 1. The results showed that the
major factor influencing biomass productivity is the light condition
at similar temperature conditions, and a higher productivity could be
achieved with higher light intensity. The similar results have been
also identified in previous research because green algae seem to
prefer high light intensity (18,19).
The daily maximum and minimum air and microalgae suspen-
sion temperatures during the cultivation experiments are shown in
Fig. 5. During the cultivation, the photobioreactor system was
cooled intermittently at noon. It can be observed that the temper-
ature of microalgae suspension followed the same tendency with
the ambient air temperature, and was lower than 35  C, which
demonstrated the microalgae suspension temperature could be
controlled by sprinkling water to the surface of the photobioreactor.
As shown in Figs. 3 and 5, the productivity was highest in the first
two batches, followed by the third and fourth batches, and the
productivity was sharply decreased in the fifth batch due to the
poor temperature lower than 5  C. The results demonstrated that
Chlorella sp. cultivation at outdoors could be carried out over a wide
range of temperatures even the culture temperature at night was
about 5  C, and the optimal air temperature at night was above
15  C (20).
Analysis of the yearly productivities of Chlorella sp. in the
pilot scale thin-film FPPBR system The thin-film FPPBR
system was located at Engebei (Ordos, China), close to the Kubuqi
FIG. 4. Environmental parameters of Engebei (Ordos, China) from April 16, 2018 to
September 30, 2018. Minimum temperature, the lowest temperature in a day;
maximum temperature, the highest temperature in a day; solar radiation, the level of
solar radiation in a day (n ¼ one replicate).
Please cite this article as: Yan, C et al., Outdoor cultivation of Chlorella sp. in an improved thin-film flat-plate photobioreactor in desertification
areas, J. Biosci. Bioeng., https://doi.org/10.1016/j.jbiosc.2019.12.007
J. BIOSCI. BIOENG.,
Desert. The time with daily lowest temperature higher than 5  C
lasted for 160 days from the middle of April to late September,
which is suitable to cultivate Chlorella sp. in the thin-film FPPBR
outdoors. The variation of solar radiation and temperature at
Engebei from the middle of April to late September of 2018 is
obtained from a Weather forecast web (http://tianqi.2345.com),
which is shown in Fig. 4. The daily lowest temperature from mid-
April to mid-May and September is lower than 15  C including
about 7 days even lower than 5  C. Low productivity was
observed in this period, such as batch 3e5, which is mainly
caused by low temperature. The days with solar radiation at
levels 4 and 5, levels 2 and 3, and level 1 in this period were
about 30 days, 19 days and 4 days, respectively. The average
productivities with solar radiation at levels 4 and 5 and levels
2and 3 in this period were estimated using the productivities of
batch 3e5, which were about 24.51 g m
2 day
1 and
8.97 g m
2 day
1. The optimal cultivating time was from the mid-
May to late August, the productivity is better because
temperature and solar radiation are more favorable, such as batch
1 and 2. In this period, the time with solar radiation at levels 4
and 5, levels 2 and 3, and level 1 were 47, 45 and 16 days,
respectively. The average productivity with solar radiation at
levels 4 and 5 and levels 2 and 3 in this period were
36.32 g m
2 day
1 and 13.01 g m
2 day
1. Therefore, the year-
round productivity of Chlorella sp. in the thin-film FPPBR would
be about 3.20 kg m
2 year
1, the year-round productivity of
Chlorella sp. in the thin-film FPPBR would be about 15.24 t ha
2
year
1, which is much higher than that in our previous research
(12). The results demonstrated that it is promising for the
economic and sustainable production of microalgae using the
photobioreactor system.
In summary, this work demonstrated the feasibility of Chlorella
sp. mass cultivation in the improved thin-film photobioreactor
system in Ordos, Inner Mongolia. The pressure drop of the FPPBR in
the improved photobioreactor system was reduced significantly,
the highest final biomass concentration and area productivity
reached 2.01 g L
1 and 49.79 g m
2 day
1, respectively. The year-
FIG. 5. Daily temperature variations of air and microalgae suspension in the photo-
bioreactor system during the cultivation experiment at 6:00 and 14:00. TA at 6:00, air
temperature at 6:00; TA at 14:00, air temperature at 14:00; TL at 6:00, microalgae
suspension temperature at 6:00; TL at 14:00, microalgae suspension temperature at
14:00 (n ¼ one replicate).
VOL. xxx, xxxx
round productivity of Chlorella sp. was estimated to be about
15.24 t ha
2 year
1 in Ordos. The results demonstrated that the
photobioreactor system was potential for mass cultivation of
microalgae in the commercial application.
ACKNOWLEDGMENTS
This work was supported by the Natural Science Foundation of
China (21706260), and the National Key R&D Program of China
(2016YFC0500904).
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