Bioprocess Biosyst Eng
DOI 10.1007/s00449-013-1058-4
ORIGINAL PAPER
Simultaneous wastewater treatment, electricity generation
and biomass production by an immobilized photosynthetic algal
microbial fuel cell
Huanhuan He • Minghua Zhou • Jie Yang •
Youshuang Hu • Yingying Zhao
Received: 2 June 2013 / Accepted: 9 September 2013
Ó Springer-Verlag Berlin Heidelberg 2013
Abstract A photosynthetic algal microbial fuel cell
(PAMFC) was constructed by the introduction of immo-
bilized microalgae (Chlorella vulgaris) into the cathode
chamber of microbial fuel cells to fulfill electricity gener-
ation, biomass production and wastewater treatment. The
immobilization conditions, including the concentration of
immobilized matrix, initial inoculation concentration and
cross-linking time, were investigated both for the growth of
C. vulgaris and power generation. It performed the best at
5 % sodium alginate and 2 % calcium chloride as immo-
bilization matrix, initial inoculation concentration of
106 cell/mL and cross-linking time of 4 h. Our findings
indicated that C. vulgaris immobilization was an effective
and promising approach to improve the performance of
PAMFC, and after optimization the power density and
Coulombic efficiency improved by 258 and 88.4 %,
respectively. Important parameters such as temperature and
light intensity were optimized on the performance. PAMFC
could achieve a COD removal efficiency of 92.1 %, and
simultaneously the maximum power density reached
2,572.8 mW/m3 and the Coulombic efficiency was 14.1 %,
under the light intensity of 5,000 lux and temperature at
25 °C.
H. He  M. Zhou  J. Yang  Y. Hu  Y. Zhao
College of Environmental Science and Engineering, Key
Laboratory of Pollution Process and Environmental Criteria,
Ministry of Education, Nankai University,
Tianjin 300071, China
H. He  M. Zhou (&)  J. Yang  Y. Hu  Y. Zhao
Tianjin Key Laboratory of Urban Ecology Environmental
Remediation and Pollution Control, Nankai University,
Tianjin 300071, China
e-mail: zhoumh@nankai.edu.cn
Keywords Photosynthetic algal microbial fuel cells 
Immobilized Chlorella vulgaris  Electricity generation
wastewater purification  Environmental factors
Introduction
Microbial fuel cells (MFCs) are a promising and self-sus-
taining technology that can recover electrical energy
directly during wastewater treatment [1–4]. In MFCs,
electrons are generated by organic matters that oxidized on
the anode, and oxygen is usually reduced to water on the
cathode. Recently, a promising biotechnology for produc-
ing electricity via living plants or microalgae has been
rapidly evolved in MFCs [5], which are photosynthesizing
and can harvest biomass, so it has potential to generate
energy in a sustainable and efficient manner [6–8].
By combining microalgae cultivation and MFCs, the photo-
synthetic algal fuel cell (PAMFC) works principally in the fol-
lowing way: microorganisms on the anode oxidize organic
matters to release protons, electrons and CO2. With light illu-
mination, the microalgae in the cathode chamber proceeds
photosynthesis, utilizing CO2 from the anode chamber as carbon
source. Meanwhile, oxygen produced from microalgae photo-
synthesis can be consumed as electron acceptor for electricity
generation. The overall biochemical reactions that occurred in
the anode chamber and the cathode chamber are as follows:
Anode chamber:
C6 H12 O6 þ 6H2 O ! 6CO2 þ 24Hþ þ 24e ð1Þ
Cathode chamber:
nCO2 þ nH2 O ! algae + light ðCH2 OÞn ðbiomassÞ
þ nO2 ð2Þ
O2 þ 4Hþ þ 4 e ! 2H2 O ð3Þ
123

Therefore, this PAMFC can accomplish energy
production and wastewater treatment without CO2
discharge due to the sequestration by microalgae. Another
advantage is that oxygen can be consumed as the cathode
electron acceptor, which overcomes the problem of oxygen
accumulation that may inhibit microalgae photosynthesis in
common photobioreactors because O2 may become toxic
when above a certain threshold [9, 10].
In more recent studies, there are few attempts on PAMFC to
fulfill carbon dioxide sequestration and electricity generation
[11–13]. Powell et al. [11] presented the growth behavior of
Chlorella vulgaris and optimized the capture of electrons.
Wang et al. [12] cultured C. vulgaris in the cathode chamber
and demonstrated that such a photosynthetic culture could act
as a biological electron acceptor on the cathode with simul-
taneous voltage output without aeration. Pandit et al. [13]
constructed a microbial carbon capture cells (MCCs) with
cyanobacteria growing in a photo-biocathode in dual-cham-
bered flat plate mediator-less MFCs, and found the power
density of the MCCs with Anabaena sparged with CO2—air
mixture (57.8 mW/m2) was much higher than that of a con-
ventional cathode sparged with air only (39.2 mW/m2).
However, the microalgae used are all in suspension sta-
tus, which is difficult to be separated from the culture media
to obtain the biomass of high value added such as biodiesel.
Besides, the microalgae tend to attach in cathode or reactor
wall, which increases the reactor internal resistance. Mic-
roalgae immobilization can solve these problems as well as
provide the advantages of high algae cell density, fast
reaction speed, strong resistant to hazardous matter, stable
and flexible operation [14]. Moreover, the immobilization
cells can also keep a high metabolic activity much longer
[15]. Therefore, the application of immobilized microalgae
as the supplier of oxygen in PAMFC might have benefits
both on microalgae separation and MFCs performance.
Therefore, in the present work, the performance of an
immobilized PAMFC was investigated by optimization of
the immobilized conditions for C. vulgaris and important
environmental factors for the process. The immobilization
conditions, such as the concentration of immobilized
matrix, initial inoculation concentration and cross-linking
time, were systematically investigated both for the growth
of C. vulgaris and power generation. Moreover, important
parameters for PAMFC including temperature and light
intensity were also studied to gain insight into the process.
Materials and methods
Preparation of immobilized algae beads
Chlorella vulgaris (FACHB 31) was purchased from
Freshwater Algae Culture Collection, Institute of
123
Bioprocess Biosyst Eng
Hydrobiology, Chinese Academy of Sciences (Wuhan,
China) and grown in a Blue-Green medium (BG11). The
concentrations of main nutrients were (unit: g/L): 1.5
NaNO3, 0.04 K2HPO4, 0.075 MgSO47H2O, 0.036
CaCl22H2O, 0.006 Citric acid, 0.006 Ferric ammonium
citrate, 0.02 NaCO3 and 1 mL trace metal solution.
The trace metal solution consisted of (unit: mg/L): 2.86
H3BO3, 1.86 MnCl24H2O, 0.22 ZnSO47H2O, 0.39
Na2MoO42H2O, 0.08 CuSO45H2O and 0.05
Co(NO3)26H2O.
The free C. vulgaris were cultivated in conical flask in
batch cultures containing BG11 and maintained under
standard conditions in an illuminating incubator with a
12:12 L:D cycle supplied by cool-white fluorescent tubes
and kept at a constant temperature of 25 ± 1 °C. Culture
pH was kept at around 7–8. All glassware was sterilized
with 75 % alcohol solution (v/v). Immobilized C. vulgaris
was prepared as previously described [16, 17]. The algae
cells in logarithmic growth phase were harvested by cen-
trifugation at 3,500 rpm for 10 min, and then they were
added into BG11 to form algal suspension. Suitable amount
of algal suspension was mixed with sodium alginate solu-
tion to yield a mixture of algal alginate suspension. This
mixture was dropped into calcium chloride solution using a
syringe to form uniform algal beads (about 3–4 mm in
size). The algal beads were left in calcium chloride solution
for certain time to harden, and then rinsed by deionized
water. In this way, the algal beads within different cell
concentrations were prepared.
PAMFC setup and operation
The PAMFC was constructed with immobilized C. vulgaris
growing in the cathode chamber of a dual-chambered
mediator-less MFCs that separated by a cation exchange
membrane (56 cm2, Ultrex CMI7000, AnkeTech Mem-
brane Separation Engineering and Technology Co., Ltd.,
China). Two identical double-chambers MFCs made of
Plexiglas were assembled by bolting two half circular
columns with each cell volume of about 200 mL. Each
chamber had a vent at the reactor top, connected through a
tube. Carbon dioxide generated from wastewater treatment
in the anode chamber was piped through the tube into the
catholyte for microalgae production via photosynthesis. It
was reported that the positive air pressure in the anode
chamber would force CO2 easily dissolved into the cath-
olyte in alkaline conditions (pH 7.2) [12].
The anode was made of graphite felt (Q-SZ03 M,
Shanghai Qiqiang Graphite Co., Ltd., China), while the
cathode was made of carbon fiber cloth (6 9 6 cm, Jilin
Shenzhou Carbon Fiber Co., Ltd., China) containing
0.1 mg/cm2 Pt catalyst. These electrodes were put in the
center of the anode chamber and cathode chamber with a
Bioprocess Biosyst Eng
distance of about 4 cm. The external resistance was fixed at
1,000 X and the anode pH was adjusted to 6.9–7.0. All
tests were operated at certain temperature in a temperature-
controlled illuminating biochemical incubator, running in a
batch mode.
After sterilized with 75 % alcohol solution, the PAMFC
reactor was filled with anolyte and catholyte in the anode
and cathode chamber, respectively. The anode chamber of
PAMFC was inoculated with anaerobic sludge (TEDA
Sewage Treatment Plant, Tianjin) and a 50 mM phosphate
buffered nutrient solution (PBS: NH4Cl 0.31 g/L, KCl
0.13 g/L, NaH2PO42H2O 3.32 g/L, Na2HPO412H2O
10.36 g/L; trace minerals 12.5 mL/L; vitamins 5 mL/L)
containing 1.0 g/L glucose as substrate. This solution was
regularly replaced by a feed solution containing glucose
(1.0 g/L) and PBS (50 mM) when the substrates were
consumed, usually within about 24 h. Normally, the num-
bers of the immobilized C. vulgaris beads that introduced
into the catholyte were 400, unless otherwise declared. The
growth phase of C. vulgaris in the cathode chamber was
relatively long, which rendered them not necessarily to be
replaced during operation. To guarantee the same light
illumination, the cathode chambers were placed at the same
angle and distance from the light source. One cycle was
ended when the voltage dropped below 100 mV.
Analysis and calculations
Cell mass was measured by optical density measurements
on a UV759 spectrophotometer (Shanghai Precise Scien-
tific Instrument Corporation, Ltd., China) at 683 nm.
Optical density was converted to cell numbers using a
previously prepared calibration curve (all cell concentra-
tions were given in cell numbers and the optical density
was repeated for three times). For determining the cell
concentration inside the immobilized beads, they were
pretreated in the following way for the subsequent optical
density determination: first dissolved in 5 mL sodium cit-
rate solution, then centrifuged at 3,500 rpm for 5 min, and
finally the cell residues were re-added into BG11 to form
algal suspension. The cell leakage was achieved by mea-
suring the optical density of culture media. For qualitative
measuring of the beads strength, the immobilized beads
were placed in a 50 mL syringe, and then imposed a certain
pressure, observing the breakage condition of beads. The
mass transfer performance of the immobilized cells could
be timely judged by the penetration depth of the methyl
orange droplets in the immobilized beads in a 100 mL
beaker.
Voltage outputs were recorded every 30 min using a
data acquisition system (PISO-813, ICPDAS Co, Ltd.).
After stable voltage outputs were observed, polarization
curves were obtained by varying the external resistances
from 1,000 to 50 X. Power density and current density
were calculated based on the effective volume of anode
chamber (anolyte volume) according to the reference [18].
The Coulombic efficiency (CE) was calculated as follows
[19, 20],
Rt2
U dt
CE ¼
t1
M ð4Þ
Rex F  b ðDCOD)V
where U is the output voltage as function of t (time), F is
Faraday’s constant (96,485 C/mol), b is the number of
electrons exchanged per mole of oxygen (=4), DCOD is the
removal of chemical oxygen demand (COD) (mg/L), Rex is
the external resistance (X), M is the molecular weight of
oxygen (=32 g/mol), and V is the wastewater volume (L).
The integral in Eq. (4) is for a time duration (t1–t2) during
which electricity is harvested through the external load.
The pH value was monitored using a desktop pH meter
(FE20, Mettler-Toledo, Switzerland). COD was measured
using the closed reflux spectrophotometric method on a
commercial COD detector (HACH, DRB 200, DR/890
Colorimeter, USA).
Results and discussion
Effect of the immobilized matrix concentration on cell
growth
Calcium alginate was used as the matrix for immobilization
of C. vulgaris. To prepare the blank beads, 3, 4, 5 %
sodium alginate (SA) and 2, 3, 5 % calcium chloride
(CaCl2) were selected. In this test, pelletization speed,
strength and mass transfer performance of beads were used
as the main indicators for the selection of matrix conditions
(Table 1). The beads prepared with 3 % SA displayed
uniform sphere configuration and good mass transfer per-
formance. However, they had very poor strength, which
made them very easily to cause the cell leakage. On the
contrary, the beads prepared with 5 % SA and 5 % CaCl2
demonstrated very good strength, but showed slow pellet-
ization speed. Therefore, 4, 5 % SA and 2, 3 % CaCl2 were
preliminarily identified as the suitable matrix conditions.
Subsequently, immobilized C. vulgaris prepared by 4, 5 %
SA and 2, 3 % CaCl2 was cultivated to investigate the cell
growth. As shown in Fig. 1, all immobilized C. vulgaris grew
well with time, and the cell numbers of the second group (5 %
SA–2 % CaCl2) reached the biggest among all groups. The
beads remained intact and cell leakage was slight (less than
0.65 9 106 cell/mL) within the stabilization period
(12 days), but all the beads were observed significant cell
leakages after 21 days. The SA concentration played an
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 Bioprocess Biosyst Eng

Table 1 Optimization of immobilized matrix concentration

Sodium alginate concentration (%) CaCl2 concentration (%) Pelletization speed and shape Strength Mass transfer performance

3 2 ??, uniform sphere ? ??

3 3 ??, uniform sphere ? ??
3 5 ??, uniform sphere ? ??
4 2 ???, uniform sphere ?? ??
4 3 ???, uniform sphere ?? ??
4 5 ??, uniform sphere ?? ?
5 2 ???, uniform sphere ??? ??
5 3 ???, uniform sphere ??? ??
5 5 ?, ellipsoidal ??? ?
? represents bad, ?? represents good, ??? represents best

14 4% SA-2% CaCl2 A 25
5% SA-2%CaCl2

cell number(106cell/ml)
12 4% SA-3%CaCl2
cell number(106cell/ml)

5% SA-3%CaCl2 20 104cell/ml
105cell/ml
10
106cell/ml
15 107cell/ml
Cell leakage (106 cell/mL)

14 12 day 140
8 21 day 5
10 cell/ml
12 120 6
10 cell/ml

cell leakage (106cell/ml)

7
100 10 cell/ml
10
6 10 80
8
60
6 40
4 5
4 20

0
2 9 12 15 18 21 24 27
2 time (d)
0
4%-2% 4%-3% 5%-2% 5%-3%
0
SA-CaCl2 0 5 10 15 20 25 30
0
0 5 10 15 20 25
time (d)
time(d)
B 700 2000

power densitty (mW/m3)

Fig. 1 Algae growth and cell leakage (insert figure) at different
sodium alginate and calcium chloride mass fractions 600
1600
voltage (mV)

500

important role in the growth of C. vulgaris. A high SA con- 400 1200

centration (5 %) not only increased the cell numbers but also
300 800
significantly decreased the cell leakage, which was less than a
half (6.6 9 106 cell/mL) of the one with 4 % SA 200
(13.4 9 106 cell/mL). A low content (2 %) CaCl2 showed no , 105 cell/ml 400
100 , 106 cell/ml
significant diversity in cell leakage but considerable benefits , 107 cell/ml
0 0
in the increase of cell numbers. This was reasonable because a 0 2 4 6 8 10 12
high osmosis pressure in the presence of high content of CaCl2 current density (A/m3)
would cause the cell activity decline. Taken the performance
of pelletization speed, strength and mass transfer into con- Fig. 2 Cell growth and cell leakage (insert figure) (a) and polar-
ization curve and power density (b) under different initial inoculation
sideration, it could be concluded that the most suitable concentrations. Conditions: SA 5 %, CaCl2 2 %, cross-linking time
immobilization matrix was 5 % SA and 2 % CaCl2. To avoid 4 h; anolyte COD 1,000 ± 52 mg/L, pH 7.20; T 25 °C; light intensity
severe cell leakage, the algae cells in logarithmic growth 1,600 lux
phase were the best choice for the subsequent experiments.
inoculation concentration was low. Therefore the effect of
Effect of initial inoculation concentration on cell initial inoculation concentration on the cell growth of C.
growth and power output vulgaris in beads was investigated (Fig. 2a). At the low
inoculation concentration of 104 cell/mL, no cell growth
Generally the final cell numbers of C. vulgaris inside the was observed. With the increase of the initial inoculation
immobilized beads would be small when the initial concentration from 105 to 107 cell/mL, the cell numbers

123
Bioprocess Biosyst Eng

also increased. The maximum cell density was observed at
106 cell/mL, though the difference among these initial
inoculation concentrations was insignificant. The insert
illustration of Fig. 2a shows the cell leakage in different
initial inoculation concentrations and culture periods. No
evident cell leakage before the 12th day was observed, but
after that, the cell leakage was gradually notable. Slight
difference on cell leakage between 105 and 106 cell/mL
initial inoculation concentrations was observed, however, a
maximum cell leakage (12 9 107 cell/mL) appeared on the
24th day on the beads inoculated with 107 cell/mL C.
vulgaris solution.
To test the power output, suitable amount of immobi-
lized beads with different initial inoculation concentrations
of C. vulgaris were put into similar PAMFC, in which the
electrochemically active microbial biofilms on the anode
surfaces were observed fully developed. And when a stable
voltage was observed, it was believed that the immobilized
C. vulgaris had carried out effective photosynthesis. After
the PAMFCs had been fully enriched, they were subjected
to experiments to obtain the polarization and power density
curves. As shown in Fig. 2b, the open circuit voltage
(OCV) of the PAMFCs with 107 cell/mL inoculation con-
centration was 656 mV, which was a little higher than that
beads had a higher activity due to a better contact with the
culture medium and light than that located in the center of
beads [22, 23]. Overall, the larger numbers of cells at the
surface of the beads caused a greater self-shading effect to
the cells inside the beads, which limited the cells growth
and increased the mass transfer resistance. This might be
explained why the power density decreased when further
increased the cell initial inoculation concentration to
107 cell/mL.
Effect of cross-linking time on cell growth and power
output
The cross-linking time is supposed to be related to the
mechanical strength of the algal beads, and the longer the
cross-linking time, the better the mechanical strength. An
inadequate cell strength results in the increase of cell
leakage, however, too high cell strength weakens the per-
meability of the algal beads and increases the mass transfer
resistance, which limits the cells growth and their
A 20
cell number(106cell/ml)

16 2h
of 105 cell/mL (637 mV) and 106 cell/mL (626 mV). 4h
6h
However, the cell voltage of the PAMFC with 106 cell/mL
12
inoculation concentration decreased less sharply than oth- 70
2h

Cell leakage (106 cell/mL)

60 4h
ers with the increase of current density. Determined by the 50
6h

steady discharging method [20], the resistance with 8 40

106 cell/mL inoculation concentration was 336 X which 30

20
was only 45 % (746 X) and 66 % (512 X) of those with 4 10

105 and 107 cell/mL inoculation concentrations, respec- 0

6 12 18 24
time (d)
tively. The maximum power densities were 505.5 ± 33.7, 0
1,811.9 ± 36.3 and 1,401.0 ± 25.3 mW/m3 at the current 0 5 10 15 20 25

densities of 2.1, 5.5, and 3.4 A/m3 in the inoculation con- time (d)
centration of 105, 106 and 107 cell/mL, respectively. The
maximum power output of the PAMFC with the inocula- B 700 2000

tion concentration of 106 cell/mL was 258 % greater than 600 power density (mW/m3)
1600
that obtained with 105 cell/mL inoculation concentration.
voltage (mV)

This difference in power performance should be related 500

1200
with the initial inoculation cell concentration since it
400
directly influenced the cell numbers inside the beads. Thus
800
a low inoculation concentration (e.g. 105 cell/mL) might 300
not produce enough oxygen via photosynthesis as electron
, 2h 400
acceptor, which led to a poor power density. However, a 200 , 4h
6h
very high inoculation concentration (e.g. 107 cell/mL) was 100
,
0
also not suitable. Hertzberg et al. found that S. costatum did 0 2 4 6 8 10
not thrive so well when immobilized in a very high initial current density (A/m3)
cell density [21]. This phenomenon could be explained that
the growing cells inside the immobilizing matrix tended to Fig. 3 Cell growth and cell leakage (insert figure) (a) and polar-
ization curve and power density (b) with different cross-linking time.
form colonies. Maximum numbers of cells per colony must
Conditions: SA 5 %, CaCl2 2 %, initial inoculation concentration
be restricted by some factors such as nutrients diffusion or 106 cell/mL; Anolyte COD 1,000 ± 52 mg/L, pH 7.20; T 25 °C;
light. Other studies showed that the cells near the surface of light intensity 1,600 lux

123

activities. Figure 3a shows the effect of cross-linking time
on the cell growth of C. vulgaris. As the cross-linking time
increased from 2 to 6 h, the order of the final cell con-
centrations from large to small was 4, 6, 2 h, and the
maximum cell numbers were 18.3 9 106, 17.3 9 106,
14.7 9 106 cell/mL, respectively. The cell leakage had no
obvious difference between 4 and 6 h cross-linking time on
the 18th day (a little smaller than that of 4 h cross-linking
time), however, they were both much less than that with
2 h cross-linking time. On the 24th day, a severe leakage
was observed in the case of 2 h cross-linking time, reach-
ing 52.2 9 106 cell/mL. These results were in accordance
with the theoretical analysis above, and confirmed that
suitable cross-linking time was vital for algae growth.
Figure 3b shows the polarization curves for different
cross-linking time. The internal resistances were deter-
mined to be 393, 336, and 498 X when the cross-linking
time was 2, 4 and 6 h, respectively. The power density for
4 h cross-linking time reached a maximum of
1,811.9 ± 36.3 mW/m3, producing a current density of
5.5 A/m3. Compared to 1,683.1 ± 40.8 mW/m3 (2 h) and
1,273.0 ± 69.4 mW/m3 (6 h), the power output was
improved by 8.5 ± 1.3 % and 43.3 ± 2.6 %, respectively.
Judged from the performance on both electricity generation
and cell growth, 4 h cross-linking time seemed to be the
most suitable.
Table 2 compares the performance before and after
optimization of the immobilized C. vulgaris in the PAM-
FC. The maximum power density after optimization was
1,811.9 ± 36.3 mW/m3, which was about 2.6 times larger
than that before optimization (505.5 ± 33.7 mW/m3). And
the maximal voltage after optimization was 505 ± 2 mV,
which was 1.6 times of that before optimization
(319 ± 3 mV). The internal resistance after optimization
(336 ± 4 X) accounted for 0.45-fold of that before opti-
mization (746 ± 5 X). Although the COD removal effi-
ciency was a little larger than that before optimization, the
Coulombic efficiency improved conspicuously by 88.4 %.
All these results proved that the optimization of immobi-
lization conditions was highly important for performance.
In PAMFC, it is well acknowledged that effective
photosynthesis and oxygen reduction are very vital to the
Table 2 Comparisons of performance before and after optimization of immobilized conditions
Resistance Plateau period Maximum power density
(X) (h) (mW/m3)
Aftera 366 ± 3 65 ± 2 1,811.9 ± 36.3
Beforeb 746 ± 5 44 ± 1.5 505.5 ± 33.7
a 6
5 % sodium alginate and 2 % calcium chloride as immobilization matrix, 4 h cross-linking time and 10 cell/mL initial inoculation
concentration
b
5 % sodium alginate and 2 % calcium chloride as immobilization matrix, 2 h cross-linking time and 105 cell/mL initial inoculation
concentration
123
Bioprocess Biosyst Eng
electricity output, which is determined by the transport of
CO2 in the cathode chamber that supposed to be a liquid
film controlled process [24]. In case of C. vulgaris
immobilized, gas produced by photosynthesis and respira-
tion promoted the beads floating on the gas–liquid inter-
face, which reduced the liquid-phase mass transfer
resistance and was favorable to the adsorption and utili-
zation of CO2. Meanwhile, oxygen could be effectively
reduced to achieve electricity generation. Thus, it was
confirmed that by optimization of the immobilized condi-
tions, the mass transfer resistance and reactor internal
resistance both decreased, which caused the enhancement
in the efficiency of electricity generation for PAMFC.
Effect of working temperature
Temperature is one of the important parameters influencing
the process performance since it affects both the growth of
microorganism on the anode and the microalgae in the
cathode chamber. Most of the single chamber MFCs adopt
30 °C as the optimal temperature [25], while the efficiency
of microalgae-based wastewater treatments is proved to be
increased with temperature from 25 to 30 °C [26]. There-
fore, in the present work, the PAMFC worked at relatively
wide temperature of 25, 30 and 35 °C was investigated,
respectively.
Figure 4 shows such effects of working temperature on
the polarization curve and power density. It was observed
that with the temperature increased from 25 to 30 °C, the
maximum power density increased somewhat from 1,811.9
to 2,072.1 mW/m3. The corresponding COD removal
efficiency was also observed slightly increased to 86.2 %.
It was reasonable since a relative high temperature would
promote the growth of microorganisms and thus the utili-
zation of the substrate [27], which would enhance the COD
removal and electron transfer to power generation. How-
ever, further increased temperature from 30 to 35 °C led to
the power density considerably decreased by 21.1 % to
1,635.1 mW/m3. At the same time, the COD removal
efficiency was found decreased to 75.5 %, while the cycle
period increased from about 175 to 213 h. These results
indicated that high temperature should have negative
Maximum voltage Coulombic COD removal
(mV) efficiency (%) efficiency (%)
505 ± 2 8.78 ± 0.2 84.7 ± 0.2
319 ± 3 4.66 ± 0.3 82.1 ± 0.4
Bioprocess Biosyst Eng

800 2100 Table 3 Comparisons of performance under different light intensity

25°C-V

power density (mW/m3)

30°C-V
700 35°C-V 1800 Light OCV Resistance Cycle COD Coulombic
25°C-P intensity (mV) (X) period removal efficiency
600 30°C-P 1500
voltage (mV)

35°C-P (lux) (h) efficiency (%)

500 (%)
1200
400
900
1,600 626 336 175 84.7 8.78
300 5,000 729 250 244 92.1 14.1
600 6,600 723 252 240 91.5 17.4
200

100 300 10,000 544 673 160 76.1 7.71

0 0
0 2 4 6 8 10
current density (A/m3)
Fig. 4 Effect of working temperature on polarization curve and
power density. Conditions: SA 5 %, CaCl2 2 %, cross-linking time
4 h; initial inoculation concentration 106 cell/mL, anolyte COD
1,000 ± 52 mg/L, pH 7.20, light intensity 1,600 lux
effects on the growth of some kinds of microorganisms on
the anode and also the growth of microalgae in the cathode
chamber. Converti et al. [28] observed similar phenomena
on C. vulgaris growth, in which the microalgae at 35 °C
displayed a 17 % decrease in growth rate when compared
with that at 30 °C, while further increase in temperature
(38 °C) led to the cells death. Based on the above out-
comes, it could be concluded that the suitable working
temperature for the process would be in range of 25–30 °C.
Effect of light intensity
Light intensity influences the growth of microalgae, which
in turn affects the performance of power output. Figure 5
shows such effects of light intensity on the polarization
curve and power density, and Table 3 lists other charac-
teristics (e.g., COD removal efficiency, Coulombic effi-
ciency) of the process under different light intensities. With
900 1600lux-V
12
the increase of light intensity from 1,600 lux, the maxi-
mum power density also increased and peaked at
2,572.8 mW/m3 under the light intensity of 5,000 lux.
Similarly, the COD removal efficiency increased from
84.7 % at 1,600 lux to 92.1 % at 5,000 lux. Correspond-
ingly, the Coulombic efficiency was also increased from
8.78 to 14.1 %. These results were logical since the mic-
roalgae growth was confirmed directly relative to the light
intensity under the autotrophic condition [29].
However, further increase the light intensity resulted in
the decline of power density. For example, at the light
intensity of 10,000 lux, the power density was only
1,123.6 mW/m3, which decreased significantly by 56.3 %
from the value at the light intensity of 5,000 lux. At the
same time, the COD removal efficiency was observed
decreased to 76.1 %, and the cycle period was also reduced
to 161 h. This was probably that a very high light intensity
led to light saturation or afterward photo-inhibition, which
harmed the growth of microalgae [30]. Another drawback
of high light intensity was the accumulation of dissolved
oxygen [29], which could be proved by the internal resis-
tance in the present work. As shown in Table 3, the internal
resistance significantly increased from 250 X at light
intensity of 5,000 lux to 673 X at light intensity of
10,000 lux. The accumulation of oxygen would increase
the transfer hindrance, which caused the resistance greatly
1600lux-P
2800 increased at the light intensity of 10,000 lux.
power density (mW/m3)

5000lux -V 5000lux-P
800 6600lux-V 6600lux-P These facts supported that light intensity was a very
2400
10000lux-V 10000lux-P
700 important environmental factor. It supported that a suitable
2000
voltage (mV)

600 light intensity (ca. 5,000–6,000 lux) would strengthen

500 1600 photosynthesis, which promoted the release of oxygen as
400 1200 the electron acceptor, and thus enhanced the power density
300 and the COD removal efficiency.
800
200
400
100
Conclusions
0 0
0 2 4 6 8 10 12
current density (A/m3) This work verified the importance of immobilization con-
ditions of C. vulgaris in PAMFC for simultaneous waste-
Fig. 5 Effect of light intensity on polarization curve and power
water treatment and electricity generation. Immobilization
density. Conditions: SA 5 %, CaCl2 2 %, initial inoculation concen-
tration 106 cell/mL, cross-linking time 4 h; anolyte COD 1,000 ± conditions including the concentration of immobilized
52 mg/L, pH 7.20, T 25 °C matrix, initial inoculation concentration and cross-linking

123

time were crucial to power generation. After optimization,
the power density and Coulombic efficiency were
improved by 258 and 88.4 %, respectively. Important
environmental factors such as light intensity and tempera-
ture were also optimized for maximizing power output in
PAMFC. It demonstrated that a suitable light intensity
(5,000–6,000 lux) and temperature (25–30 °C) would
benefit the simultaneous wastewater treatment, electricity
generation and biomass production.
Acknowledgments This work was financially supported by Natural
Science Foundation of China (No.51178225, No. 21273120 and No.
21250110515), GEFC09-12, Fund for the Doctoral Program of Higher
Education of China (20110031110025), NCET-08-0296, SRF for
ROCS, SEM (2009-1001) and Open Fund of Key Laboratory of
Original Agro-environment Quality of Ministry of Agriculture and
Tianjin Key Laboratory of Agro-environment and Agro-product
Safety (2012-SZJJ-CYM).
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