CN102399723B - Bacillus with electrogenesis characteristic and application thereof in microbiological fuel cell - Google Patents
Bacillus with electrogenesis characteristic and application thereof in microbiological fuel cell Download PDFInfo
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- CN102399723B CN102399723B CN2011103475263A CN201110347526A CN102399723B CN 102399723 B CN102399723 B CN 102399723B CN 2011103475263 A CN2011103475263 A CN 2011103475263A CN 201110347526 A CN201110347526 A CN 201110347526A CN 102399723 B CN102399723 B CN 102399723B
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E60/00—Enabling technologies; Technologies with a potential or indirect contribution to GHG emissions mitigation
- Y02E60/30—Hydrogen technology
- Y02E60/50—Fuel cells
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Abstract
A bacillus with electrogenesis capability is characterized in that the Bacillus pumilus MFC-2 is preserved in China General Microbiological Culture Collection Center on 12th, July 2011 and has a preservation number CGMCC No.5056. The Bacillus pumilus MFC-2 bacterium colony in regular circle has smooth surface, luster and tidy edge, and is in opaque lacte; a thallus is in thin rod shape, straight, round on two ends and Gram-positive, and is generally 0.6-0.7 mum*2.0-3.0 mum. The MFC-2 bacterial strain of the invention can utilize a plurality of carbon sources and grow under anaerobic and aerobic conditions. The microbiological fuel cell prepared by utilizing the bacterial strain has high electrogenesis capability; the bacterial strain can be directly used for electrogenesis in the microbiological fuel cell and can utilize organic matters in sewage to generate electricity.
Description
Technical field
The invention belongs to biological technical field, particularly a kind of genus bacillus be bacillus pumilus (
Bacillus pumilus) the electrogenesis characteristic and the applied research in microbiological fuel cell.
Background technology
Along with the mankind's progress and development, further strong to the demand of the energy.In order to solve the energy shortage problem, the mankind are actively seeking the novel energy mode, and the development and utilization renewable energy source has become the important component part of the world energy sources strategy of sustainable development.Microbiological fuel cell (Microbial Fuel Cells, MFCs) receives investigator's concern in recent years as a kind of novelty sewage disposal and energy recovery technology that integrates sewage purification and electrogenesis.MFCs is a class special in the fuel cell, refer to utilize enzyme or microorganism as catalyzer, by its metabolism, under normal temperature, normal pressure, anaerobism, the chemical energy of the large weight organic compounds that contains in sanitary sewage or the trade effluent is converted into the device of electric energy, comprise two utmost point chambers of negative and positive, the centre is separated by proton exchange membrane, reclaims clean energy in degradation of organic substances.
Aspect wastewater treatment, MFCs has not available several the advantages of existing method.At first, MFCs can be used as power supply and carries out the energy reparation.Secondly, under stable condition, MFCs processes lacking of the more aerobic processing of excess sludge that produces.During the 3rd, MFCs processed, microorganism only consumed a small amount of pollutent and provides energy for its growth, and most of pollutent is converted into electric energy.The technology that MFCs is considered to a very promising production clean energy and carries out the environment-friendly type wastewater treatment.
The key factor that affects the MFCs effect is the electrogenesis microorganism.At present, if the electrogenesis microbial host Proteobacteria of separation and the bacterium of Firmicutes mostly are facultative anaerobe.Patent of the present invention relate to a kind of bacillus pumilus (
Bacillus pumilus), this bacterial classification has very strong electricity generation performance, and the application aspect microbiological fuel cell there is not yet report both at home and abroad.
Summary of the invention
The purpose of this invention is to provide from nature and isolate the bacillus pumilus with high reactivity electricity generation ability, described bacillus pumilus (Bacillus sp.) MFC-2 bacterial strain (hereinafter to be referred as the MFC-2 bacterial strain), under be categorized as genus bacillus, be preserved in " China Committee for Culture Collection of Microorganisms's common micro-organisms " center "; address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City; postcode: 100101; preservation date is on July 12nd, 2011, and preserving number is CGMCC No.5056.
Another object of the present invention provides the application of MFC-2 bacterial strain aspect microbiological fuel cell, and this bacterial strain has electricity generation ability by force and can utilize organic waste water to carry out the performance of electrogenesis.Microbiological fuel cell with its preparation has broad application prospects.
MFC-2 bacterial strain of the present invention is the Bacillus strain that reported first has the electrogenesis characteristic in this kind, has very high electrogenesis activity in microbiological fuel cell, has wide prospects for commercial application.
MFC-2 bacterial strain of the present invention is to separate in Nanjing University's Life Science College organic waste water in front of the door, through microscopic examination, staining reaction, conventional physiological and biochemical property experiment and the 16s rDNA sequential analysis of bacterium colony and form, this identification of strains be bacillus pumilus (
Bacillus pumilus) the MFC-2 bacterial strain, have following characteristics:
Bacterium colony rule circle, smooth surface, glossy, opaque oyster white, neat in edge;
The thalline thin rod shape, thalline is straight, two ends are circular, Gram-positive, is generally 0.6 ~ 0.7 μ m * 2.0 ~ 3.0 μ m;
Biochemical reactions shows as: anaerobic growth experiment, V-P reaction experiment, 7%NaCl growth experiment, gelatine liquefication experiment, hippurate are tested, the casein hydrolysising experiment is all positive; Indoles experiment, phenylalanine decarboxylase activity experiment, malonate utilization experiment, nitrate reduction experiment, Citrate trianion utilization are tested all negative; Glucose, pectinose, N.F,USP MANNITOL, wood sugar can be utilized, but starch can not be utilized;
16s rDNA the sequencing results shows, this bacterium with
Bacillus aerophilus,
Bacillus subtilis,
Bacillus altitudinisWith
Bacillus pumilusHomology be 100%.In conjunction with the biochemical reactions feature, identify its be bacillus pumilus (
Bacillus pumilus).
MFC-2 bacterial strain of the present invention can utilize several kinds of carbon source, all can grow under the anaerobic and aerobic condition; In microbial fuel cell unit, has very high electricity generation ability.The report that bacillus pumilus has electricity generation performance belongs to first.
The application of MFC-2 bacterial strain of the present invention: (1) this bacterium can be directly used in as electron donor with glucose and carry out electrogenesis in the microbiological fuel cell; (2) can in Sewage Environment, utilize organic components to carry out electrogenesis; (3) this bacterium can be with absorption or the mode on the electrode holder of being embedded in as the anode of microbial fuel cell catalyzer.
Description of drawings
Fig. 1 is the organigram of microbiological fuel cell;
Fig. 2 is that the MFC-2 bacterial strain is at the scanning electron microscope (SEM) photograph that adheres on carbon paper electrode surface;
Fig. 3 is the open circuit voltage of MFC-2 bacterial strain in microbiological fuel cell-time curve schematic diagram.
Embodiment
Screening, separation, the evaluation of embodiment 1:MFC-2 bacterial strain
In the natural waste water in the Nanjing University campus, get 500 mL nature waste water and put into the anolyte compartment of microbiological fuel cell, microbial fuel cell unit adopts traditional H type double-chamber structure (Fig. 1), formed by two vials, between with the Glass tubing docking that is connected on the vial, middle Nafion 211 proton exchange membrane that adopt E.I.Du Pont Company, two Room are clamped and couple together with clip, electrode adopts high-purity carbon paper, wire connects with copper wire, the part of exposing gums down, and external circuit connects the fixed resistor of 1000 Ω, and the potential difference at resistance two ends adopts volt ohm-milliammeter to measure.Galvanic anode passes into high pure nitrogen to guarantee anaerobic state, and negative electrode adopts potassium ferricyanide solution.Whole cell apparatus is placed in constant temperature (25 ℃) room operation, until external circuit voltage be elevated to stable after, the voltage of surveying after the week is 600 mV.Under anaerobic environment, the carbon felt anode of lattice in the anolyte compartment is put into physiological saline, rock, the thalline of stick on top is entered in the physiological saline, and renewed vaccination again moves battery in the fresh anolyte and carries out enrichment, and triplicate can both reach the sample of very high voltage, carry out the strain separating purifying at anaerobic culture box, substratum is LB, obtains the stronger bacterial strain of a strain electricity generation ability---MFC-2, and the MFC-2 bacterial strain is seen Fig. 2 at the scanning electron microscope (SEM) photograph that adheres on carbon paper electrode surface.Adopt the glycerine preserving process for subsequent use in-80 ℃ of refrigerator preservations.
16s rDNA the sequencing results is carried out maximum homology relatively in GenBank, find 16s rDNA sequencing result with
Bacillus aerophilus,
Bacillus subtilis,
Bacillus altitudinisWith
Bacillus pumilusHomology be 100%, according to " identification mark (table 1) of above-mentioned four kinds of genus bacillus of describing in uncle's Jie Shi handbook and " the common bacteria identification handbook " has been measured biochemical reactions and the growth characteristics of MFC-2 bacterial strain, sees Table 1.
The biochemical reactions of table 1. MFC-2 and growth characteristics measurement result
Annotate :+: the positive ,-: feminine gender, d: difference, ND: do not survey
The cultivation of embodiment 2:MFC-2 bacterial strain
The MFC-2 bacterial strain bacterium colony of picking activation from the LB flat board, be inoculated in the 15mL centrifuge tube that contains 5mL liquid LB substratum, 37 ℃, 120 rev/mins, cultivate after 12 hours, with 1% inoculum size it is joined and contain 250mL electrogenesis substratum, as the 500mL Erlenmeyer flask in, consisting of of electrogenesis substratum: glucose (10g/L), yeast extract (5g/L), NaHCO
3(10g/L), NaH
2PO
4(8.5g/L), 37 ℃, 120 rev/mins, to cultivate 12 hours, the centrifugal bacterial strain that again obtains is used for electrogenesis research.
The detection of embodiment 3:MFC-2 bacterial strain electrogenesis
Cultivation and inoculation method according to example 2 statements, from 200ml with glucose as obtaining the growth logarithm cell in late period the substratum of electron donor, centrifugal (8000rpm, 10min), the gained precipitation is re-dispersed in the anaerobic culture medium of 50mM glucose as electron donor after washing three times with 50mM PBS damping fluid, is inoculated in the anode of microbiological fuel cell.The logical high pure nitrogen of anode of microbial fuel cell liquid keeps anaerobic environment, and catholyte is that the potassium ferricyanide solution of 0.1M PBS damping fluid is as electron acceptor.Cell apparatus is as described in the example 1, and the cathode and anode chamber leads to 25 ℃ of thermostat(t)ed waters, concerns over time with CHI660D electrochemical workstation record battery open circuit voltage, and the result as shown in Figure 3.
Claims (1)
1. bacillus pumilus with electricity generation ability is characterized in that: described bacillus pumilus (
Bacillus pumilus)Called after MFC-2 has been preserved on July 12nd, 2011 that " China Committee for Culture Collection of Microorganisms's common micro-organisms " center ", its preserving number are CGMCC No. 5056.
2. the application of bacillus pumilus according to claim 1 in microbiological fuel cell.
3. the application of bacillus pumilus according to claim 2 in microbiological fuel cell is characterized in that: described bacillus pumilus can be directly used in as electron donor with glucose and carry out electrogenesis in the microbiological fuel cell; Perhaps can in Sewage Environment, utilize organic components to carry out electrogenesis.
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| CN103337652B (en) * | 2013-06-14 | 2015-09-02 | 国家电网公司 | A kind of fuel cell |
| CN103337651B (en) * | 2013-06-14 | 2015-08-26 | 国家电网公司 | A kind of Biological-agent fuel cell |
| CN114982413B (en) * | 2022-06-16 | 2023-12-08 | 南京农业大学 | Solid state fermentation preparation and application of biochar-based bacillus inoculant |
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| CN101570731A (en) * | 2009-03-25 | 2009-11-04 | 新奥科技发展有限公司 | Method for domesticating and separating electricigens by electrochemistry |
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