CN101552340B - 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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CN101552340B
CN101552340B CN2009100979887A CN200910097988A CN101552340B CN 101552340 B CN101552340 B CN 101552340B CN 2009100979887 A CN2009100979887 A CN 2009100979887A CN 200910097988 A CN200910097988 A CN 200910097988A CN 101552340 B CN101552340 B CN 101552340B
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fuel cell
compartment
cathode
liquid
marine yeast
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CN101552340A (en
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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 bacteriasolution 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 organism is converted into electric energy by the microorganism catalysis reaction, 2007), its mechanism is that the anode organism decomposes the generation proton under the oxygenizement of microorganism, 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 intestinal bacteria substratum subsequently.After the discovering in microbiological fuel cell of the eighties in 20th century adds the electron transport intermediate, current density and output rating 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 compatibility is strong, can effectively move under cold condition; Operation does not produce waste gas, is beneficial to environmental protection; Have the widespread 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 organism that directly is meant is on electrode in the oxidation, and electronics directly is transferred to electrode from organism; Be meant that indirectly the electronics that the organism 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 metallorganics, toluylene red (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 microorganism can't independently carry out electron transport, have only and itself have electron transport intermediate (Zhuwei Du et al. in only a few microorganism cells or its meta-bolites, 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 level, 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 reference 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 wastewater 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.Biofuel cell---progress and application.Biosensor and biological electronics, 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 toluylene red is as the microorganism growth 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 toluylene red as microbiological fuel cell electrogenesis electrodes transfer body.Use biochemistry and biotechnology, 2000,66 (4): 1292-1297.)
[7]U.Schroder,Juliane?Nieβen,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 think 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 taint 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; It is IMI 394084 that Tallman protects minus sign, 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 anolyte compartment and the cathode compartment that opens wide, in the anolyte compartment, be provided with anode electrode, in cathode compartment, be provided with cathode electrode, between anolyte compartment and cathode compartment, proton exchange membrane is set, the organic-fuel liquid that contains bacterium liquid is set in the anolyte compartment, 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 anolyte compartment 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 compartment is made according to the ratio that every 100ml deionized water adds 0.04~0.05g Tripotassium iron hexacyanide.
Further improvement as microbiological fuel cell of the present invention: the volume of solution in the volume=cathode compartment of the organic-fuel liquid in the anolyte compartment.
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 substratum of sterilising treatment is housed with the bacterial classification of transfering loop 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 anolyte compartment, 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 Tripotassium iron hexacyanide and deionized water are packed into according to the ratio of 0.04~0.05g:100ml form solution in the cathode compartment;
5), with in the organic-fuel liquid in the end immersion anolyte compartment of anode electrode, in the solution in the end immersion cathode compartment (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 TW20 9TY 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 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 level 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 raw material consumption 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, and life-time service also guarantees the marine yeast activity.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 anolyte compartment and cathode compartment sealing also can adapt to anaerobic species to generate electricity through transforming, the environmental compatibility of device is strong.Except proton, other small molecules can't see through, and have guaranteed glucose, NaH between cathode compartment and the anolyte compartment in the selected proton exchange membrane energy assurance system 2PO 4, the Tripotassium iron hexacyanide 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 synoptic diagram of Fig. 1;
Fig. 3 is Rhodosporidium paludigenum Fell ﹠amp; The Tallman fuel cell passes through the resulting output voltage graphic representation 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 anolyte compartment 2 and the cathode compartment 1 that opens wide, the capacity=216ml of the capacity=cathode compartment 1 of anolyte compartment 2.Anolyte compartment 2 and cathode compartment 1 are processed into by synthetic glass, anolyte compartment 2 and cathode compartment 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 having on the infiltrative micropore ceramics diaphragm-operated two sides, dry back and getting.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 anolyte compartment 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 compartment 1, be provided with potassium ferricyanide solution; In anolyte compartment 2, be provided with anode electrode 5, in cathode compartment 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 work:
Preparation NYDA substratum:
Contain nutrient broth 8g, yeast powder 5g, glucose 10g, agar 20g and sodium-chlor 10~150 grams in every 1L NYDA substratum, all the other are water.Prepare back 121 ℃ of sterilization 20min.
Preparation NYDB substratum:
Contain nutrient broth 8g, yeast powder 5g, glucose 10g and sodium-chlor 10~150 grams in every 1LNYDB substratum, all the other are water.Prepare back 121 ℃ of sterilization 20min.
In deionized water, add glucose, be mixed with the glucose solution that glucose concn 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 substratum of 60ml and pack in the container after the sterilising treatment, then with transfering loop 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 anolyte compartment 2 of proton exchange membrane 3 and cathode compartment 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 anolyte compartment 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 anolyte compartment 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 compartment 1, add Tripotassium iron hexacyanide 0.1g, mend to cathode compartment 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 anolyte compartment 2 of anode electrode 5, in the solution in the end immersion cathode compartment 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 reometer, and then obtain output rating.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 anolyte compartment 2 begin generating after, the resistance of varistor 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.Fs, this moment, external resistance was 1000 Ω from beginning by about 150 minutes; After subordinate phase 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.
Output rating is obtained by the data of output voltage and electric current.Similarly, the variation of external resistance directly affects output rating.Can be got by Fig. 4, the variation tendency of output rating 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 peak 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. the purposes of a marine yeast is characterized in that: described marine yeast Rhodospridium paludigenum Fell﹠amp; The Tallman preserving number is IMI 394084, can be used in the microbiological fuel cell direct generation of electricity.
2. microbiological fuel cell, comprise unlimited anolyte compartment (2) and the cathode compartment (1) that opens wide, in anolyte compartment (2), be provided with anode electrode (5), in cathode compartment (1), be provided with cathode electrode (4), it is characterized in that: between anolyte compartment (2) and cathode compartment (1), proton exchange membrane (3) is set, the organic-fuel liquid that contains bacterium liquid is set in anolyte compartment (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 Nation-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 anolyte compartment (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 compartment (1) is made according to the ratio that every 100ml deionized water adds 0.04~0.05g Tripotassium iron hexacyanide.
5. microbiological fuel cell according to claim 4 is characterized in that: the volume of the interior solution of volume=cathode compartment (1) of the organic-fuel liquid in described anolyte compartment (2).
6. microbiological fuel cell according to claim 5 is characterized in that: be provided with anode electrode (5) in described anolyte compartment (2), be provided with cathode electrode (4) in cathode compartment (1); Anode electrode (5) material is an iron, and cathode electrode (4) material is a graphite.
7. as the preparation method of any one microbiological fuel cell in the claim 2~6, it is characterized in that 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 substratum of sterilising treatment is housed with the bacterial classification of transfering loop 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 anolyte compartment (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 Tripotassium iron hexacyanide and deionized water are packed into according to the ratio of 0.04~0.05g: 100ml form solution in the cathode compartment (1);
5), with in the organic-fuel liquid in the end immersion anolyte compartment (2) of anode electrode (5), in the solution in the end immersion cathode compartment (1) of cathode electrode (4).
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CN105543115B (en) * 2016-01-22 2020-03-13 中国科学院天津工业生物技术研究所 Marine yeast and application thereof in bioethanol production
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