Bioresource Technology 109 (2012) 308311

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

Production of algal biomass (Chlorella vulgaris) using sediment microbial fuel cells
Hyeon Jin Jeon a, Kyu-won Seo a, Sang Hyun Lee a, Yung-Hun Yang a, Rangarajulu Senthil Kumaran a, Sunghyun Kim b, Seok Won Hong c, Yong Su Choi c, Hyung Joo Kim a,
a

Department of Microbial Engineering, Konkuk University, Seoul 143-701, Republic of Korea Department of Bioscience and Biotechnology, Konkuk University, Seoul 143-701, Republic of Korea c Water Environment Center, Korea Institute of Science and Technology, P.O. Box 131, Cheongryang, Seoul 130-650, Republic of Korea
b

a r t i c l e

i n f o

a b s t r a c t
In this study, a novel algal biomass production method using a sediment microbial fuel cell (SMFC) system was assessed. Under the experimental conditions, CO2 generation from the SMFC and its rate of increase were found to be dependent on the current generated from the SMFC. However, the CH4 production rate from the SMFC was inhibited by the generation of current. When Chlorella vulgaris was inoculated into the cathode compartment of the SMFC and current was generated under 10 X resistance, biomass production from the anode compartment was observed to be closely associated with the rate of current generation from the SMFC. The experimental results demonstrate that 420 mg/L of algae (dry cell weight) was produced when the current from the SMFC reached 48.5 mA/m2. Therefore, SMFC could provide a means for producing algal biomass via CO2 generated by the oxidation of organics upon current generation. 2011 Elsevier Ltd. All rights reserved.

Article history: Available online 16 June 2011 Keywords: Sediment microbial fuel cell Algal cultivation Chlorella vulgaris Bioelectrochemical CO2 generation Bioelectrochemical CH4 suppression

1. Introduction Recently, a signicant amount of attention has been focused on renewable energy sources that might help satisfy the everincreasing demand for energy consumption and environmental protection (Choi et al., 2010; De Schamphelaire and Verstraete, 2009). Among possible renewable energy sources, microalgae have gained increased attention as they contain a signicant quantity (e.g. 2050% dry cell weight) of neutral lipids for biodiesel production (Hu et al., 2008). Various methods for the cultivation of algae have been proposed ever since Harder and von Witsch proposed the mass cultivation of diatoms to produce urgently needed fat during World War II (Liang et al., 2009). In recent years, signicant efforts have expended toward the development of biological aspects (i.e. strain selection and/or improvement, etc.) and non-biological aspects (i.e. light source, fermentor design, cultivation method, etc.) for use in algal production (Brennana and Owendea, 2010). Despite recent efforts to improve the production yield of algal biomass, there remain some technological barriers to overcome before the economic production of a stable energy source becomes feasible. Among these technological barriers, CO2 is one of the critical factors in photosynthesis, along with light, water, and nutrients. Particularly in the case of high-density cultures of algal biomass in a

Corresponding author. Tel.: +82 2 2049 6111; fax: +82 2 446 2677.
E-mail address: hyungkim@konkuk.ac.kr (H.J. Kim). 0960-8524/$ - see front matter 2011 Elsevier Ltd. All rights reserved. doi:10.1016/j.biortech.2011.06.039

bioreactor, effective CO2 supply is recognized as a crucial factor. In the case of open pond-type cultures such as raceway-type ponds or lakes, quality control, slow growth, and low CO2 partial pressure constitute major obstacles (Brennana and Owendea, 2010). Along with the problems listed above, the presence of organic bottom lake sediment is associated with the generation of CH4, which has serious greenhouse gas effects. To remove CH4 generation from conventional lake or swamp sediment, a variety of methods, including ultrasonication, O2 feeding, heat treatment, and acid/base treatment, have been proposed for the treatment of sediment (Elbeshbishya et al., 2010). In particular, it has been reported that the addition of inorganic electron acceptors to sediment inhibits CH4 generation (Scholten and Stams, 1995). In a previous study, a microbial fuel cell (MFC) using lake sediment as an electron donor for electrochemically active bacteria was examined, and it was determined that this method could be used successfully to reduce the organic matter content of the sediment (Hong et al., 2008). In the present study, we demonstrate a novel algal production system using a sediment microbial fuel cell (SMFC) capable of decreasing CH4 generation capability of organic rich-lake sediment, which was divided into two parts. First, using organic-rich sediment obtained from a natural lake, correlations between current generation and gas (CO2 and CH4) production in the SMFC were established. Secondly, the algal production capability of the SMFC and the applicability of algal culture for SMFC operation without any external oxidant were investigated.

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2. Methods 2.1. Algal strain, medium, and sediment for SMFCs The Chlorella vulgaris strain (KCTC AG10002) was provided by the Korean Culture Type Collection (Daejun, Korea). The medium for cultivation of C. vulgaris was Bolds Basal Medium (BBM, pH 6.8) (Lima et al., 2010). The cells were grown in 250 ml Erlenmeyer asks on an orbital shaker set at 150 rpm at 25 C under constant orescence light. Sediment for the construction of SMFCs was obtained from Ilgam lake, an articial lake at Konkuk University, Seoul, and a typical hypereutrophic lake in Korea (collection date: June 2010). The average amounts of organic matter in the sediment as measured by the loss on ignition (LOI) and readily oxidizable organic matter (ROOM) methods were 10.4% LOI and 3.52% ROOM, respectively (Hong et al., 2008). 2.2. SMFC construction Fig. 1 shows a schematic diagram of the sediment microbial fuel cell (SMFC) employed in this study. The SMFC consisted of an anode and cathode positioned at opposite sides of a poly-acrylic plastic cylindrical chamber (d = 110 mm, h = 250 mm, t = 2 mm). Both the anode (d = 80 mm, t = 2 mm) and cathode (d = 90 mm, t = 2 mm) consisted of graphite felt (GF series, GEE graphite Ltd., UK). Electrodes were connected externally with concealed copper wire. The sediment (775 g wet weight) was initially added to the chamber (ca. 50 mm in thickness), and the anode was placed in the middle of the sediment layer. The sediment was then covered with sterilized sand (638 g wet weight). For direct collection of the gas generated from the anode compartment (i.e. sediment), a funnel type gas collector (d = 80 mm) was placed on the surface of the sediment of the SMFC and connected to a gas sample bag (232 series, SKC, USA). Four sets of the SMFC system were used in this study. 2.3. SMFC operation The SMFCs were operated in a uorescence light (light intensity of 81 lmol/m2 s) incubator at constant temperature of 25 C. To investigate the effect of external resistance on current and gas generation, the cathode compartment was initially lled with BBM, and the cathode was aerated at a rate of 200 ml/min. The connec-

tion between the anode and cathode was made via external load resistors. BBM containing 1% glucose (w/v) as a fuel for operation of the SMFCs was supplied to the system (0.04 ml/min) with a peristaltic pump. When the system set-up was completed, the current generated from each SMFC was monitored with a digital multimeter (model 2000, Keithley, USA) (Choi et al., 2010). The gas generated during operation (15 days) of the SMFCs was collected via the gas collector and stored in the sample bag. Sampling for bicarbonate analysis was made via the sampling port (Fig. 1). For algae cultivation in the SMFCs, the gas collector in the cathode compartment was removed and the precultured algae inoculated (20% v/v) into the cathode compartment. During algal cultivation, aeration to the cathode was stopped. Cultivations were carried out for 15 days in the SMFCs with different load resistors. To prevent bacterial contamination, the SMFCs were irradiated with UV light (kmax = 204 nm) eight times a day (10 s every 3 h at 200 lmol/m2 s intensity) (Seo et al., 2009). All experiments were conducted at least in duplicate.

2.4. Analysis Gaseous CO2 and CH4 generated from the SMFCs were analyzed via gas chromatography as described previously (Holland et al., 1999). CO2 dissolved in the cathode compartment was measured via the acid/base titration based on the bicarbonate concentration (Fresenius et al., 1988). The dry weight of the algal culture sample was determined by drying 50 ml of the algal suspension at 80 C in a drying oven for 24 h after ltration through pre-dried and preweighted 0.45 lm lter paper.

3. Results and discussion 3.1. Effect of external resistance on generation of current, CO2, and CH4 After the SMFCs with different external load resistors were set up, electrical current and gas generation from the SMFCs were monitored. The current gradually increased over 15 days, upon which it achieved steady state. Fig. 2 shows the relationships among electrical current generation, CO2 generation, and CH4 generation from the SMFCs connected with different external load resistors.

Fig. 1. Schematic diagram of the algal culture system using SMFC.

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CH4

10
CO2 HCO3
-

30

Current density (mA/m )

Current density

25 20 15 10

2
5 0
10 100 500 Open circuit

External resistance ()
Fig. 2. Effect of external resistance on the generation of CO2 and CH4 from the SMFCs. The data were evaluated after 15 days of SMFC operation with different external load resistors.

The algal biomass density in the SMFC connected to a 10 X resistor was about 420 mg/l, which is approximately 1.5-fold higher than that in a conventional bioreactor (315 mg/l at 23 days; Liang et al., 2009). In SMFCs with high-level load resistors (i.e. 500 X) or open circuit conditions, however, the biomass concentrations in the cathode compartments of the SMFCs were lower (230 mg/l in 500 X and 120 mg/l in open circuit) compared to that observed in the 10 X resistance SMFC. In this experiment, the dependency of algal concentration on current generation from the SMFC may be attributed to the rate of CO2 generation based on bacterial activities in the anode compartment. It should be noted that one of the weak points of algal cultivation is the requirement of an external CO2 supply. Therefore, the results of this study imply that the cultivation of algal species in an SMFC is indeed possible without the external addition of CO2. 3.3. Operation of SMFC in the presence of algae culture Continuous supplementation of an oxidant (such as oxygen) to the cathode is essential to maintain the potential and current of a MFC (Choi et al., 2010). Under illumination, assuming that algae in the cathode compartment generate oxygen, the rate of current generation of an SMFC will be proportional to the algal oxygenproducing activity, if oxygen is rate-limiting. Fig. 3a shows the current generation pattern from the SMFC in the presence of the algal culture. As a control experiment, an SMFC without algal inocula-

Under open circuit conditions (i.e. no electrical current ew through the SMFC), about 1.78 mM CO2 (gaseous CO2 + bicarbonate) was obtained. When the electrodes were connected via a 10 X external load resistor (current: 33 mA/m2), however, the maximum concentration of CO2 reached 8.83 mM. The experimental results demonstrate that the amount of CO2 generated from the SMFC was related closely to the rate of current generation from the SMFC. Previous studies have shown that current generation from a microbial fuel cell is completely dependent on carbon metabolism by the bacteria, and the anode of the microbial fuel cell acts as an electron acceptor for electrochemically active bacteria (Kim et al., 2002). Therefore, the correlation between increased CO2 generation and increased current generation from SMFCs with low level external load resistors can be attributed to the increased metabolic speed of the electrochemically active bacteria in the sediment. In the case of a high level of resistance (i.e. 500 X or open circuit), limited electron ow suppresses the metabolic activity of the electrochemically active bacteria, resulting in a low level of CO2 generation. Electrical current also changed the rate of CH4 generation from the SMFC. A gradual decrease in the rate of CH4 generation occurred when the current from the SMFC was increased. Under open circuit conditions, about 1.34 mM CH4 was produced. When the SMFC was connected to a 10 X external load resistor, the CH4 concentration was reduced to 0.13 mM. Previous research using a MFC demonstrated that electron loss is attributable to the activity of the anode (i.e. electron acceptor), as a low level external load resistor reduced the rate of CH4 generation (Hong et al., 2008). In this experiment, changes in the gas generation rate from the SMFC can be attributed to the action of the anode in the sediment, with oxidation of the electron donor and subsequent variations in the activity of related microorganisms (Aelterman et al., 2008). Taking these observations into consideration, additional experiments were conducted to investigate the possibility of algal cultivation in the SMFC using CO2 generated from the system (i.e. anode compartment) as an inorganic carbon source. 3.2. Effect of external resistance on the growth of algae in the SMFC To evaluate the algal culture capability of the SMFC, algae-inoculated SMFCs connected with different external load resistors were operated. When the current generation and biomass concentration in the SMFCs were monitored after 15 days of operation, we found that the connection of a low-level load resistor to the SMFC induced both higher values of current and biomass concentration.

HCO3 , CO2 & CH4 production (mM)

A
Current density (mA/m )
2

50
with algae, without air without algae, with air

40

30

20

10

10

12

14

16

Time (day)

B
Current density (mA/m )
2

60 50 40 30 20 10 0 16

18

20

22

Time (day)
Fig. 3. Operation of SMFCs under various experimental conditions. (A) Comparison of currents generated from SMFCs with and without algal cultivation. (B) Current generation pattern from an SMFC with and without illumination (lights-on: 18 h, lights-off: 6 h, arrows indicate lights-off times).

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tion was operated with air. In the control experiment, the general form of the amperometric response to the connection of the circuit was a rapid rise in current as a function of time, followed by exponential decay to a plateau value (ca. 33 mA/m2). In the case of the algae-inoculated SMFC with continuous illumination and no air supply, a relatively slow increase in current was observed during the early stage of operation (09 days). However, after 11 days of operation, the current from the SMFC increased at a high rate and reached a steady state (48 mA/m2). One possible reason for this higher current is that the increased amount of O2 produced by the algae under illumination interacted with the SMFC cathode. As is shown in Fig. 3b, when illumination was subjected to an on off cycle (light: 18 h, dark: 6 h), the rate of current generation from the SMFC also uctuated. These results show that the oxygen generated from the algae could be directly employed as an oxidant for the SMFC cathode. Therefore, the application of algal cultivation to a MFC format has a synergistic effect on both algal growth and SMFC operation. In this study, the rapid reduction of oxygen at the surface of the cathode may have induced a reduction in the concentration of oxygen, which probably stimulated algal growth in the SMFC (Paerl and Pinckney, 1996). Current research by our group is focused on the optimization of algal growth and algal lipid accumulation/quantication for biodiesel production (Park et al., 2010). 4. Conclusions C. vulgaris was successfully cultivated in an SMFC format without any further addition of CO2. It was determined that the amount of CO2 and CH4 production from the anode compartment was dependent on current generation from the SMFC. When C. vulgaris was inoculated into the cathode compartment of the SMFC, increases in both current and algal concentration were observed. These ndings show that the synergic interaction between algal culture and SMFC; in other words, CO2 generated by bacterial activity is consumed by algal cultivation, and the O2 produced from the algae is consumed by the SMFC cathode for current generation. Acknowledgement This subject is supported by the Korea Ministry of Environment as a Converging Technology Project (201-101-007).

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