CN101552340A - Uses of a marine yeast and corresponding microorganism fuel cell and method for preparing the same - Google Patents

Uses of a marine yeast and corresponding microorganism fuel cell and method for preparing the same Download PDF

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CN101552340A
CN101552340A CNA2009100979887A CN200910097988A CN101552340A CN 101552340 A CN101552340 A CN 101552340A CN A2009100979887 A CNA2009100979887 A CN A2009100979887A CN 200910097988 A CN200910097988 A CN 200910097988A CN 101552340 A CN101552340 A CN 101552340A
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fuel cell
chamber
anode chamber
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liquid
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郑晓冬
殷赟
刘宜胜
王一非
余挺
李伟
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Zhejiang University ZJU
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Abstract

The invention discloses uses of a marine yeast, preservation No. of the marine yeast Rhodosporidimu paludigenum Fell & Tallman is IMI 394084, the marine yeast can be used in microorganism fuel cell for electricity generation directly. The invention discloses a microorganism fuel cell at the same time, including an open anode chamber (2) with an anode electrode (5) provided inside and an open cathode chamber (1) with a cathode electrode provided inside (4), a proton exchange membrane is provided between the anode chamber (2) and the cathode chamber (1), a liquid organic fuel containing bacteria solution is provided in the anode chamber (2), and the bateria solution is obtained by fermenting the marine yeast of preservation No. 394084. The invention also discloses method of preparing the microorganism fuel cell at the same time. The microorganism fuel cell of the invention is provided with features of environment friendly.

Description

A kind of purposes of marine yeast and corresponding microorganism fuel cell and preparation method
Technical field
The present invention relates to a kind of purposes of marine yeast.Also relate to the microbiological fuel cell that utilizes this marine yeast gained and the preparation method of this battery.
Background technology
Microbiological fuel cell is device (the Z.Du et al. that the chemical energy in the organic substance is converted into electric energy by the microorganism catalysis reaction, 2007), its mechanism is that the anode organic substance decomposes the generation proton under the oxidation of microbe, be passed to negative electrode through proton exchange membrane, form the loop to reach electrogenesis effect (R.M.Allen et al., 1993) with external circuit.The notion of microbiological fuel cell is proposed in 1910 by Potter the earliest, and he has realized MFC electrogenesis the earliest with platinum electrode in the Escherichia coli culture medium subsequently.After the discovering in microbiological fuel cell of the eighties in 20th century adds the electron transport intermediate, current density and power output all improve (F.Davis et al. on a large scale, 2007), provide possible chance (K.Rabaey et al., 2005) for practical application.It is compared with conventional fuel cell, and possess some advantages: raw material is extensive, can directly be converted into electric energy efficiently; Environmental suitability is strong, can effectively move under cryogenic conditions; Operation does not produce waste gas, is beneficial to environmental protection; Have the extensive use potentiality in the area that lacks power infrastructures,, can satisfy more multi-form demand (K.Rabaey et al., 2005) as a kind of new forms of energy.At present, problems such as energy scarcity and environmental pollution obtain paying attention to day by day, and the whole world is more and more strong to the demand of novel renewable energy, and the microbiological fuel cell research field is emerged rapidly, becomes the international research focus.
According to the difference of electron transport mode, microbiological fuel cell can be divided into direct microbiological fuel cell and indirectly microbiological fuel cell (F.Davis et al .2007).The so-called organic substance that directly is meant is on electrode in the oxidation, and electronics directly is transferred to electrode from organic substance; Be meant that indirectly the electronics that the organic substance reaction is produced needs to be delivered on the electrode by the electron transport intermediate.Typical electronic is transmitted intermediate and is comprised pigment and metallorganic, dimethyl diaminophenazine chloride (D.H.Park et al., 1999,2000) for example, methylene blue (U.Schroder et al., 2003), thionine (Y.Choi et al., 2003) etc.But the electron transport intermediate costs an arm and a leg, and needs often to replenish, and relative cost is high; And poisonous mostly, make it can not in open environment, use (Feng Yali etc., 2005).
Because most of microbe can't independently carry out electron transport, have only and itself have electron transport intermediate (Zhuwei Du et al. in only a few microbial cell or its metabolite, 2007), can not add external source electron transport intermediate and carry out the direct generation of electricity.Therefore the discovery of this quasi-microorganism has solved the global problem that necessary interpolation toxicity external source electron transport intermediate just can obtain higher output power, is to the important breakthrough in the microbiological fuel cell research process.
So far the bacterial classification of having reported that carries out direct electrogenesis mainly comprises Shewanella putrefaciens (corrupt Shiva Salmonella) (H.J.Kim et al., 2002), Geobacter sulfurreducens (sulphur reduction earth bacillus) (D.R.Bond et al., 2003), Geobacter metallireducens (B.Min et al., 2005) and Rhodoferax ferrireducens (S.K.Chaudhuri et al., 2003), stable operation, have good electricity generation performance, but above bacterial classification does not all relate to marine yeast.
Rhodosporidium paludigenum Fell﹠amp; Tallman is the domestic marine yeast that obtains that separates first, and in international agricultural of Britain international standard institute of microbiology and genetic resources preservation center, biological center (CABI Genetic ResourceCollection) preservation, preserving number is IMI 394084.It has very high low temperature resistant and salt tolerance, belongs to facultative marine yeast, and low for all general bacterial classification of requirement of pH, temperature, oxygen content, environment-adapting ability is strong; Determine true border non-toxic type (Wang Yifei etc., 2008) through food-grade chmice acute per os toxicity test.
The list of references of above mentioning is specific as follows:
[1] Z.Du, H.Li, T Gu.2007.A state of the art review on microbial fuel cells:A promisingtechnology for wastewater treatment and bioenergy.Biotechnology Advances 25:464-482. (Du Zhuwei, Li Haoran, Gu Tingyue.Microbiological fuel cell summary: a kind of promising waste water treatment and bioenergy new technology.The biotechnology progress, 2007,25:464-482.)
[2] R.M.Allen, H.P.Bennetto.1993.Microbial fuel-cells:electricity production fromcarbohydrate.Applied Biochemistry and Biotechnology, 39/40:27-40. (Alan, Benny is logical.Microbiological fuel cell: carbohydrate electrogenesis.Use biochemistry and biotechnology, 1993,39/40:27-40.)
[3] F.Davis, S.P.J.Higson.2007.Biofuel cells-Recent advances and applications.Biosensorsand Bioelectronics 22:1224-1235. (Davis, Xi Gesen.Bio-battery---progress and application.Biology sensor and bioelectronics, 2007,22:1224-1235.)
[4] K.Rabaey, W.Verstraete.2005.Microbial fuel cells:novel biotechnology for energygeneration.Trends in Biotechnology.23 (6): 291-298. (sieve shellfish, Fan Site.Microbiological fuel cell: production capacity new technology.Biotechnology trend, 2005,23 (6): 291-298.)
[5] D.H.Park, M.Laivenieks, M.V.Guettler, M.K.Jain, J.G.Zeikus.1999.Microbial Utilization of Electrically Reduced Neutral Red as the Sole Electron Donor forGrowth and Metabolite Production.Applied and environmental microbiology, 65 (7): 2912-2917. (Parker, La Weike, Gothic, letter, Aix-en-Provence.Electricity degraded dimethyl diaminophenazine chloride is as the growth of microorganism and the metabolism of electron donor separately.Use biochemistry and biotechnology, 1999,65 (7): 2912-2917.)
[6] D.H.ParK, J.G.Zeikus.2000.Electricity Generation in Microbial Fuel Cells Using NeutralRed as an Electronophore.Applied and environmental microbiology, 66 (4): 1292-1297. (Parker, Aix-en-Provence.With dimethyl diaminophenazine chloride as microbiological fuel cell electrogenesis electrodes transfer body.Use biochemistry and biotechnology, 2000,66 (4): 1292-1297.)
[7] U.Schroder, Juliane Nie β en, Fritz Scholz.2003.A Generation of Microbial Fuel Cells withCurrent Outputs Boosted by More Than One Order of Magnitude.Angew.Chem., 115:2986-2989. (Si Gaode, Ni Fen thinks deeply this.The electric current of microbiological fuel cell is produced the method for carrying forward vigorously.Germany's applied chemistry, 2003,115:2986-2989.)
[8] Y.Choi, E.Jung, S.Kim, Seunho Jung.2003.Membrane fluidity sensoring microbial fuelcell.Bioelectrochemistry, 59:121-127. (Cai, ginger, gold, Jiang.Membrane flow sensing microbiological fuel cell.Bioelectrochemistry, 2003,59:121-127.)
[9] Feng Yali, Lian Jing, Zhang Wenming, Zhou Liang, Zhu Xueyuan, a kind of medium-free microbial fuel cell, 2007, CN 1889297A.
[10] H.J.Kim, H.S.Park, M.S.Hyun, I.S.Chang, M.Kim, B.H.Kim.2002.A mediatorlessmicrobial fuel cell using a metal reducing bacterium, Shewanella putrefaciens.EnzymeMicrobiology Technology, 30:145-152. (gold, Parker, Korea Spro, normal, gold.A kind of with the microbiological fuel cell of metallic reducing bacterial putrefaction west watt Salmonella as no amboceptor.Microbial technique, 2002,30:145-152.)
[11] D.R.Bond, D.R.Lovley.Electricity Production by Geobacter sulfurreducens Attachedto Electrodes.2003.Applied and Environmental Microbiology, 69 (3): 1548-1555. (Bond, Lai Fuli.To be adsorbed on the Geobacter sulfurreducens electrogenesis on the electrode.Use and environmental microorganism 2003,69 (3): 1548-1555.)
[12] B.Min, S.Cheng, B.E.Logan.Electricity generation using membrane and salt bridgemicrobial fuel cells.2005.Water Res, 39:1675-1686. (bright, become the Lip river root.Carry out the production by biological method for electrically with film and salt bridge.Water resources research, 2005,39:1675-1686.)
[13] S.K.Chaudhuri, D.R.Lovley.Electricity generation by direct oxidation of glucose inmediatorless microbial fuel cells.2003.Nature Biotechnology, 21:1229-1232. (prosperous moral Harry, Lai Fuli.Direct oxidation glucose carries out the medium-free microbial fuel cell electrogenesis.Nature Biotechnol, 2003,21:1229-1232.)
[14] Wang Yifei, 2008, the doctorate paper, marine yeast is to the research of diseases biological control of postharvest fruits and vegetables.
Summary of the invention
The technical problem to be solved in the present invention provides a kind of purposes of marine yeast, and the bacterium liquid of this marine yeast fermentation back gained can be used for making microbiological fuel cell.
In order to solve the problems of the technologies described above, the invention provides a kind of purposes of marine yeast, this marine yeast Rhodospridium paludigenum Fell﹠amp; The Tallman preserving number is IMI 394084, can be used for the microbiological fuel cell direct generation of electricity.
The present invention also provides the bacterium liquid making that utilizes this marine yeast fermentation back gained and the microbiological fuel cell that gets simultaneously, comprise unlimited anode chamber and the cathode chamber that opens wide, in the anode chamber, be provided with anode electrode, in cathode chamber, be provided with cathode electrode, between anode chamber and cathode chamber, proton exchange membrane is set, the organic-fuel liquid that contains bacterium liquid is set in the anode chamber, and bacterium liquid is that preserving number is the bacterium liquid of the marine yeast fermentation back gained of IMI 394084; Proton exchange membrane is provided with the infiltrative micropore ceramics diaphragm of having of Nafion-117 coating for the surface.
Improvement as microbiological fuel cell of the present invention: contain bacterium liquid 9～10ml, glucose 0.45～0.5g, NaH in the organic-fuel liquid in every 100ml anode chamber 2PO 40.05～0.075g, all the other are deionized water.
Further improvement as microbiological fuel cell of the present invention: the solution in the cathode chamber is made according to the ratio that every 100ml deionized water adds 0.04～0.05g potassium ferricyanide.
Further improvement as microbiological fuel cell of the present invention: the volume of solution in the volume=cathode chamber of the organic-fuel liquid in the anode chamber.
Further improvement as microbiological fuel cell of the present invention: anode electrode material is an iron, and cathode electrode material is a graphite.
The present invention also provides the preparation method of microorganism fuel cell simultaneously, may further comprise the steps:
1) be that the marine yeast of IMI 394084 inserts in the NYDA medium slant 26～28 ℃ of activation 24～48 hours with preserving number;
2), insert in the container that 30～70ml NYDB culture medium of sterilization treatment is housed with the bacterial classification of oese after with above-mentioned activation, constant temperature was cultivated 8～16 hours for 26～28 ℃ 150～250 rev/mins, collected bacterium liquid;
3) with above-mentioned bacterium liquid, glucose, NaH 2PO 4Packing into deionized water forms the organic-fuel liquid that contains bacterium liquid in the anode chamber, contains bacterium liquid 9～10ml, glucose 0.45～0.5g, NaH in every 100ml organic-fuel liquid 2PO 40.05～0.075g, all the other are deionized water.
4), the potassium ferricyanide and deionized water are packed into according to the ratio of 0.04～0.05g: 100ml form solution in the cathode chamber;
5), with in the organic-fuel liquid in the end immersion anode chamber of anode electrode, in the solution in the end immersion cathode chamber (1) of cathode electrode.
Marine yeast (the Rhodosporidium paludigenum Fell﹠amp that the present invention is selected; Tallman) be preserved in Britain international standard fungal studies international agricultural of institute and genetic resources preservation center, biological center (International Mycological Institute, CABI Genetic Resource Collection), preservation address: international agricultural in Beckham road, the Britain prefecture Ai Ge of TW209TY Surrey town and biological center Britain center, preservation date: 2006.4.19, deposit number: IMI 394084.This bacterial strain is disclosed in application number is 200610155209.0 patent.The bacterium colony pinkiness, smooth surface can be grown in NYDB that is added with 1%～15%NaCl concentration and NYDA.
The marine yeast Rhodospridium paludigenum Fell﹠amp that utilizes of the present invention; The microbiological fuel cell that Tallma makes has following advantage:
1. the bacterial classification of the present invention's use is the domestic marine yeast Rhodospridium paludigenumFell﹠amp that obtains that separates first; Tallman identifies preservation through international agricultural of Britain international standard institute of microbiology and genetic resources preservation center, biological center (CABIGenetic Resource Collection), and preserving number is IMI 394084.It has very high low temperature resistant and salt tolerance, belongs to facultative marine yeast, for pH, and temperature, all general bacterial classification of the requirement of oxygen content is low, and environment-adapting ability is strong; Determine true border non-toxic type through food-grade chmice acute per os toxicity test, safe in utilization pollution-free; Therefore microbiological fuel cell environmental protection of the present invention.
2. the marine yeast Rhodospridium paludigenum Fell﹠amp that uses of the present invention; Tallman has very Johnson ﹠ Johnson head's ability, and cell proliferation is rapid, can arrive the electrogenesis peak fast and keep stablely, and the cycle of operation is long, and consumption of raw materials is few.
3. the marine yeast Rhodospridium paludigenum Fell﹠amp that uses of the present invention; Tallman, can under the condition that does not add electronics transmission intermediate, carry out the direct generation of electricity and reach higher electricity generation ability, be that the only a few reported in the world up to now can not add that electronics transmits intermediate and unique marine yeast in the bacterial classification of the direct generation of electricity, has very strong adaptive capacity to environment, applied range.
4. microbial fuel cell unit of the present invention is easy to operate, can add bacterial classification and raw material as required, uses and guarantee the marine yeast activity for a long time.Can accomplish scale production, to reach the purpose of practical application.
5. microbiological fuel cell of the present invention is simple in structure, and cost of manufacture is cheap, is special by the generating of aerobic bacterial classification and facultative bacterial classification, and as making anode chamber and cathode chamber sealing also can adapt to anaerobic species to generate electricity through transforming, the environmental suitability of device is strong.Except proton, other micromolecule can't see through, and have guaranteed glucose, NaH between cathode chamber and the anode chamber in the selected proton exchange membrane energy assurance system 2PO 4, the potassium ferricyanide can't traverse to another chamber from a chamber.
Description of drawings
Below in conjunction with accompanying drawing the specific embodiment of the present invention is described in further detail.
Fig. 1 is the structural representation of microbiological fuel cell of the present invention;
Fig. 2 is the actual test mode schematic diagram of Fig. 1;
Fig. 3 is Rhodosporidium paludigenum Fell﹠amp; The Tallman fuel cell passes through the resulting output voltage curve chart of changes in resistance from the beginning electrogenesis to electrogenesis in about 15 hours;
Fig. 4 is Rhodosporidium paludigenum Fell﹠amp; The Tallman fuel cell is from beginning electrogenesis to electrogenesis about 15 hours, the output power curve figure that obtains by changes in resistance.
Embodiment
Embodiment 1, Fig. 1 have provided a kind of microbiological fuel cell, comprise unlimited anode chamber 2 and the cathode chamber 1 that opens wide, the capacity=216ml of the capacity=cathode chamber 1 of anode chamber 2.Anode chamber 2 and cathode chamber 1 are processed into by polymethyl methacrylate, anode chamber 2 and cathode chamber 1 rely on proton exchange membrane 3 to realize separating, proton exchange membrane 3 adopts following method to make: all apply one deck Nafion-117 solution on the two sides with infiltrative micropore ceramics diaphragm, get after the drying.Has the Nafion series membranes that infiltrative micropore ceramics diaphragm can select for use DuPont company to produce.The organic-fuel liquid that contains bacterium liquid is set in anode chamber 2, and this bacterium liquid is that preserving number is the bacterium liquid of the marine yeast fermentation back gained of IMI 394084; In cathode chamber 1, be provided with potassium ferricyanide solution; In anode chamber 2, be provided with anode electrode 5, in cathode chamber 1, be provided with cathode electrode 4.Anode electrode 5 is fabricated from iron, and cathode electrode 4 is made by graphite.
The preparation method of microorganism fuel cell may further comprise the steps:
1), preparation:
Preparation NYDA culture medium:
Contain nutrient broth 8g, dusty yeast 5g, glucose 10g, agar 20g and sodium chloride 10～150 grams in every 1L NYDA culture medium, all the other are water.Prepare back 121 ℃ of sterilization 20min.
Preparation NYDB culture medium:
Contain nutrient broth 8g, dusty yeast 5g, glucose 10g and sodium chloride 10～150 grams in every 1LNYDB culture medium, all the other are water.Prepare back 121 ℃ of sterilization 20min.
In deionized water, add glucose, be mixed with the glucose solution that concentration of glucose is 0.1g/ml.In deionized water, add NaH 2PO 4, be mixed with NaH 2PO 4Concentration is the NaH of 0.05g/ml 2PO 4Solution.
2), with preserving number be the marine yeast Rhodospridium paludigenum Fell﹠amp of IMI 394084; Tallman inserts in the above-mentioned NYDA medium slant 27 ℃ of activation 36 hours;
3), get the above-mentioned NYDB culture medium of 60ml and pack in the container after the sterilization treatment, then with oese with step 2) bacterial classification inserts in the said vesse after the activation of gained, constant temperature was cultivated 10 hours for 27 ℃ 200 rev/mins, collected bacterium liquid;
4), will be fixed with the anode chamber 2 of proton exchange membrane 3 and cathode chamber 1 is 30% H successively in volumetric concentration 2O 2, deionized water, 0.5mol/L H 2SO 4And respectively boiled 1 hour in the deionized water, be kept in the deionized water standby then.
Anode electrode 5 and cathode electrode 4 all adopt following method to clean: soak in the HCl of 1mol/L solution earlier to remove foreign ion, in the NaOH of 1mol/L solution, soak then and remove the cell of its surface adsorption, living contaminants is avoided in last all ultraviolets sterilization 6 hours.
The bacterium liquid 20ml that gets the step 3) gained inserts in the anode chamber 2, adds the 0.1g/ml glucose solution 10ml of step 1) gained in the anode chamber 2,0.05g/ml NaH 2PO 4Solution 3ml mends to the anode chamber 2 with deionized water then and is have one's bosom filled with (promptly being settled to 216ml with deionized water); Formation contains the organic-fuel liquid of bacterium liquid.
5), in cathode chamber 1, add potassium ferricyanide 0.1g, mend to cathode chamber 1 with deionized water then and be have one's bosom filled with (promptly being settled to 216ml) with deionized water.
6), with in the organic-fuel liquid in the end immersion anode chamber 2 of anode electrode 5, in the solution in the end immersion cathode chamber 1 of cathode electrode 4.
Test 1, adopt the described method of Fig. 2 to detect embodiment 1 described microbiological fuel cell, anode electrode 5 is connected by variable rheostat 6 with cathode electrode 4, record voltage according to the digital multimeter 7 that is connected variable rheostat 6 two ends then; Can also record electric current simultaneously by inserting ammeter, and then obtain power output.These anode electrode 5 materials are iron, and diameter is 0.1cm, and surface area is 6.3cm 2Cathode electrode 4 is unpolished high purity graphite, and physical surface is long-pending to be 16cm 2
Under the condition of 15 ℃ of room temperatures, with Rhodosporidium paludigenum Fell﹠amp; Tallman inserts anode chamber 2 begin generating after, the resistance of rheostat 6 is transferred to 2000 Ω from 200 Ω, voltage data and current data begin to be gathered.
As shown in Figure 3, Rhodosporidium paludigenum Fell﹠amp; In nearly 15 hours of the Tallman electrogenesis,, obtain the output voltage curve by changes in resistance.Phase I, this moment, external resistance was 1000 Ω from beginning by about 150 minutes; After second stage rose to 2000 Ω with resistance, output voltage significantly improved, and stable continuing to about 500 minutes changes external resistance to 200 Ω once more, and output voltage obviously descends, and only maintains about 0.2V.By the conversion external resistance, obtaining maximum output voltage in this scope is 0.9371V.
Power output is obtained by the data of output voltage and electric current.Similarly, the variation of external resistance directly affects power output.Can be got by Fig. 4, the variation tendency of power output is consistent with the variation tendency of output voltage, and the peak power output of whole process can obtain when external resistance is 2000 Ω, and the external resistance that be described this moment is near the internal resistance of this microbiological fuel cell.Conclusion can get: the internal resistance of this microbiological fuel cell is at least greater than 2000 Ω, and can obtain internal resistance of cell value simultaneously by changing external resistance to obtain the peak power output of this battery at 2000 Ω environs.Whole process maximum power obtains about battery operation to 500 minute, is 0.4592mW.This microbiological fuel cell can drive the miniature electric components and parts flexibly.
At last, it is also to be noted that what more than enumerate only is a specific embodiment of the present invention.Obviously, the invention is not restricted to above embodiment, many distortion can also be arranged.All distortion that those of ordinary skill in the art can directly derive or associate from content disclosed by the invention all should be thought protection scope of the present invention.

Claims (7)

1, a kind of purposes of marine yeast is characterized in that: described marine yeast Rhodospridium paludigenum Fell ﹠amp; The Tallman preserving number is IMI 394084, can be used for the microbiological fuel cell direct generation of electricity.
2, a kind of microbiological fuel cell, comprise unlimited anode chamber (2) and the cathode chamber (1) that opens wide, in anode chamber (2), be provided with anode electrode (5), in cathode chamber (1), be provided with cathode electrode (4), it is characterized in that: between anode chamber (2) and cathode chamber (1), proton exchange membrane (3) is set, the organic-fuel liquid that contains bacterium liquid is set in anode chamber (2), and described bacterium liquid is that preserving number is the bacterium liquid of the marine yeast fermentation back gained of IMI 394084; Described proton exchange membrane (3) is provided with the infiltrative micropore ceramics diaphragm of having of Nafion-117 coating for the surface.
3, microbiological fuel cell according to claim 2 is characterized in that: contain bacterium liquid 9～10ml, glucose 0.45～0.5g, NaH in the organic-fuel liquid in every 100ml anode chamber (2) 2PO 40.05～0.075g, all the other are deionized water.
4, microbiological fuel cell according to claim 3 is characterized in that: the solution in the cathode chamber (1) is made according to the ratio that every 100ml deionized water adds 0.04～0.05g potassium ferricyanide.
5, microbiological fuel cell according to claim 4 is characterized in that: the volume of the interior solution of volume=cathode chamber (1) of the organic-fuel liquid in described anode chamber (2).
6, microbiological fuel cell according to claim 5 is characterized in that: described anode electrode (5) material is an iron, and cathode electrode (4) material is a graphite.
7,, it is characterized in that may further comprise the steps as the preparation method of any one microbiological fuel cell in the claim 2～6:
1) be that the marine yeast of IMI 394084 inserts in the NYDA medium slant 26～28 ℃ of activation 24～48 hours with preserving number;
2), insert in the container that 30～70ml NYDB culture medium of sterilization treatment is housed with the bacterial classification of oese after with above-mentioned activation, constant temperature was cultivated 8～16 hours for 26～28 ℃ 150～250 rev/mins, collected bacterium liquid;
3) with above-mentioned bacterium liquid, glucose, NaH 2PO 4Packing into deionized water forms the organic-fuel liquid that contains bacterium liquid in the anode chamber (2), contains bacterium liquid 9～10ml, glucose 0.45～0.5g, NaH in every 100ml organic-fuel liquid 2PO 40.05～0.075g, all the other are deionized water.
4), the potassium ferricyanide and deionized water are packed into according to the ratio of 0.04～0.05g: 100ml form solution in the cathode chamber (1);
5), with in the organic-fuel liquid in the end immersion anode chamber (2) of anode electrode (5), in the solution in the end immersion cathode chamber (1) of cathode electrode (4).
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