CN103131651A - Bacillus subtilis bacterial strains and application thereof in microbial power generation - Google Patents
Bacillus subtilis bacterial strains and application thereof in microbial power generation Download PDFInfo
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- CN103131651A CN103131651A CN2013100456946A CN201310045694A CN103131651A CN 103131651 A CN103131651 A CN 103131651A CN 2013100456946 A CN2013100456946 A CN 2013100456946A CN 201310045694 A CN201310045694 A CN 201310045694A CN 103131651 A CN103131651 A CN 103131651A
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- 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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- 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
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Abstract
The invention relates to the field of microbial technology, in particular to bacillus subtilis bacterial strains. The bacterial strain is MMR-1, and the preservation number of the bacterial strain is CGMCC No.6871. The invention further provides a microbial fuel cell which comprises an anode chamber, a cathode chamber, and an external circuit, wherein the anode chamber comprises anode solutions, an anode, and electricity generating microbes, and the electricity generating microbes are biological pure cultures of the bacillus subtilis bacterial strains or biological pure cultures which have all the identifying characteristics of the bacillus subtilis bacterial strains. The bacillus subtilis bacterial strains are facultative anaerobes and have a function of generating electricity, so that the rang of electricity generating microbes is expanded, and anaerobic experiment conditions can be improved through the preparation of the microbial fuel cell by using the bacillus subtilis bacterial strains.
Description
[technical field]
The present invention relates to microbial technology field, be specifically related to a kind of new bacillus subtilis strain and the application in microbiological fuel cell and electricity generation by microorganism thereof.
[background technology]
Increasingly serious along with problems such as global warming and fossil oil exhaustion, the research of the alternative energy and renewable energy source is subject to increasing attention.In recent years, a kind ofly directly chemical energy can be converted into the power generation assembly of electric energy---microbiological fuel cell (microbial fuel cell, MFC) with its distinctive advantage, becomes the study hotspot in emerging energy field gradually.MFC combines microbial technique with battery technology, utilize microorganism as catalyzer, the chemical energy in organism to be converted into the power generation assembly of electric energy, has generating and waste treatment double effects.Except high concentrated organic wastewater, MFC can also be with the generating that acts as a fuel of the pollutents such as sanitary sewage.The ultimate principle of MFC is: organism acts as a fuel in the anaerobism anolyte compartment by microbiological oxidation, the electronics that produces is by microorganism-capturing and pass to galvanic anode, electronics arrives negative electrode by circuit, thereby form the loop generation current, and proton arrives negative electrode by exchange membrane, with the oxygen water generation reaction.
MFC power density ratio other types fuel battery power density is low, and its research at present also is in laboratory level or lab scale level.The electricity generation performance of MFC is subject to the impact of many factors, comprises composition and the pH value of electrolytic solution, electrode area size and interelectrode distance, proton exchange membrane size and classification etc., especially the biological catalyst type that adopts of anolyte compartment, namely electrogenesis microorganism type, more remarkable on the impact of its electricity generation performance.The kind that studies show that the anolyte compartment bacterium as Logan etc. is different, the power density that MFC produces is different (Logan B also, Cheng S, Watson V. Graphite fiber brush anodes for increased power production in air-cathode microbial fuel cells.Environ.Sci.Technol, 2007,41 (9): 3341-3346.).Therefore, the electricity generation performance of raising electrogenesis microorganism becomes problem demanding prompt solution.
Thereby bacterium is transmitted electronics to the anode generation current by adding intermediate or self producing intermediate, only has at present minority about self generating the electrogenesis research of the outer electrogenesis bacterium of mesosome born of the same parents, wherein mainly concentrate on two class alienation metallic reducing bacterium (Shewanella and Geobacter), all belong to strictly anaerobic bacterium, harsh to requirement for experiment condition, still have larger gap from large-scale application.Have no the report that facultative anaerobe has the electrogenesis activity in prior art.
[summary of the invention]
The technical problem to be solved in the present invention is to improve the electricity generation performance of electrogenesis microorganism, and improves the experimental anaerobic condition.
For this reason, the invention provides a kind of bacillus subtilis strain (Bacillus subtilis), described bacterial strain is MMR-1, depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation date: on November 22nd, 2012, preserving number: CGMCC No.6871.
Further, the 16S rRNA gene order of described bacterial strain is SEQ ID No.1.
The invention provides a kind of biology pure growth of bacterium strain, having preserving number is all identification marks of the bacillus subtilis strain MMR-1 of CGMCC No.6871.
The present invention provides above-mentioned bacillus subtilis strain or the application of above-mentioned biology pure growth in electricity generation by microorganism in addition.
The present invention also provides a kind of microbiological fuel cell, and the electrogenesis microorganism of described microbiological fuel cell is above-mentioned bacillus subtilis strain or above-mentioned biology pure growth.
Preferably, the electron donor of described electrogenesis microorganism is the organism in glucose, sodium acetate, N.F,USP MANNITOL or sewage.
Preferably, described microbiological fuel cell comprises anolyte compartment, cathode compartment and external circuit three parts, and described anolyte compartment comprises anolyte, anode and electrogenesis microorganism.
Preferably, described anolyte is for adding blue-green algae substratum, broth culture or the sewage of carbon source.
Preferably, the negative electrode of described microbiological fuel cell is the air cathode of Pt/C catalysis.
Bacillus subtilis strain of the present invention is facultative anaerobe, has electricity generation performance, has enlarged the scope of electrogenesis microorganism, simultaneously, utilizes described bacillus subtilis strain to prepare microbiological fuel cell and can improve the experimental anaerobic condition.As anolyte, described subtilis is decomposed the organism in sewage, has not only realized chemical energy is converted into electric energy, has realized simultaneously the dirty water decontamination handles with sewage.
[description of drawings]
Fig. 1 is embodiment 1 Anodic carbon paper scanning electron microscope (SEM) photograph.
Fig. 2 is MMR-1 bacterial strain phylogeny tree graph.
Fig. 3 is microbiological fuel cell voltage and the power stage temporal evolution figure of embodiment 2.
[embodiment]
In order to make purpose of the present invention, technical scheme and advantage clearer, below in conjunction with the drawings and specific embodiments, the present invention is described in further detail.Should be appreciated that specific embodiment described herein only in order to explain the present invention, is not intended to limit the present invention.
The invention provides a kind of bacillus subtilis strain (Bacillus subtilis), described bacterial strain is MMR-1, subtilis MMR-1 of the present invention separates from the blue-green algae nutrient solution, depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation date: on November 22nd, 2012, preserving number: CGMCC No.6871.
Further, described identification of strains feature is as follows: the bacterium colony surface irregularity is opaque, and dirty white has the gauffer projection; It is shaft-like that microscopically is; Gramstaining is positive; Facultative anaerobe; Can utilize the materials such as glucose, N.F,USP MANNITOL, starch, protein as carbon source; Nitrate reduction experiment, formation indoles are tested all positive; Phenylalanine deaminase is tested, the V-P reaction experiment is negative.
Further, the 16S rRNA gene order of described bacterial strain is SEQ ID No.1.The 16S rRNA gene order of described bacterial strain in the Genebank database without identical sequence.
Bacillus subtilis strain of the present invention is facultative anaerobe, has electricity generation performance, has enlarged the scope of electrogenesis microorganism.
The present invention provides a kind of biology pure growth of bacterium strain on the other hand, and having preserving number is all above-mentioned identification marks of the bacillus subtilis strain MMR-1 of CGMCC No.6871.
The present invention also provides a kind of microbiological fuel cell on the other hand, and the electrogenesis microorganism of described microbiological fuel cell is above-mentioned bacillus subtilis strain or above-mentioned biology pure growth.
Further, described microbiological fuel cell comprises anolyte compartment, cathode compartment and external circuit three parts, and the anolyte compartment comprises anolyte, anode and electrogenesis microorganism, and described electrogenesis microorganism is above-mentioned bacillus subtilis strain or above-mentioned biology pure growth.
Simultaneously, utilize described bacillus subtilis strain or its pure growth to prepare microbiological fuel cell and can improve the experimental anaerobic condition.
Embodiment 1
The separation of bacterial strain MMR-1
Microbiological fuel cell adopts two chamber H-type ampuliform system, is 1.6cm with the unorganic glass bottle that covers of two 250ml by diameter
2Glass tubing is connected, and covers the unorganic glass bottle respectively as anolyte compartment and cathode compartment with two, as electrode, is inoculated into the anolyte compartment with blue-green algae as anode catalyst with carbon paper, and carbon paper is separately fixed at cathode compartment and anolyte compartment by graphite fixture, and two interelectrode distances are 12cm
2, be connected with external resistance or volt ohm-milliammeter by copper cash.
Microbiological fuel cell through the reaction, on the anode carbon paper enrichment one deck microbial film.The anode carbon paper is placed under scanning electron microscope observes, as shown in Figure 1, microbial film mainly is comprised of bacterium.With repeatedly streak inoculation of the microbial film on the anode carbon paper, obtain the purifying bacterial strain, obtained strains is screened, obtain having the bacterial strain of electricity generation performance, the strain that wherein electricity generation performance is the highest is bacterial strain MMR-1.
The evaluation of bacterial strain MMR-1
Obtained strains MMR-1 is carried out respectively identification of morphology, Physiology and biochemistry evaluation and Molecular Identification, and result is as follows:
The identification of morphology result: the bacterium colony surface irregularity is opaque, and dirty white has the gauffer projection; It is shaft-like that microscopically is; Gramstaining is positive;
Physiology and biochemistry qualification result: facultative anaerobe; Can utilize the materials such as glucose, N.F,USP MANNITOL, starch, protein as carbon source; Nitrate reduction experiment, formation indoles are tested all positive; Phenylalanine deaminase is tested, the V-P reaction experiment is negative.
The Molecular Identification result: the 16S rRNA gene order of described MMR-1 bacterial strain is SEQ ID No.1.In ncbi database with the BLAST retrieval close sequence with MMR-1 bacterial strain 16S rRNA gene, MMR-1 bacterial strain 16S rRNA gene order, the close sequence that retrieves and close division bacteria group represented sequence analysis, phylogenetic tree construction, as shown in Figure 2, wherein, the content in bracket is the accession number of each bacterial strain in ncbi database; Numerical value is this branch node supporting rate, and the MMR-1 bacterial strain is in separately a different branch, illustrates that the MMR-1 bacterial strain is new bacterial strain.
According to above qualification result, the MMR-1 bacterial strain belongs to subtilis, and it is carried out preservation, and preservation information is as follows: depositary institution is China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation date is on November 22nd, 2012, and preserving number is CGMCC No.6871.
The strictly anaerobic bacterium that is that has the electrogenesis activity in prior art, the MMR-1 bacterial strain that the present embodiment separates belongs to facultative anaerobe.Utilize described bacillus subtilis strain or its pure growth to prepare microbiological fuel cell and can improve the experimental anaerobic condition.
Embodiment 2
Embodiments of the invention 2 provide a kind of microbiological fuel cell, comprise anolyte compartment, cathode compartment and external circuit three parts, the anolyte compartment comprises anolyte, anode and electrogenesis microorganism, and described electrogenesis microorganism is embodiment 1 gained MMR-1 bacterial strain or the biology pure growth with all identification marks of MMR-1 bacterial strain.
Further, anolyte is the mixed solution of 195ml BG11 substratum and sodium acetate, and anode is carbon paper, and negative electrode is the air electrode of Pt/C catalysis, contains concentration 50mM phosphoric acid buffer in cathode compartment.
When the anolyte in the present embodiment also can be broth culture or sewage, particularly sewage as anolyte, also can decompose the organism in sewage except generating electricity, not only realize chemical energy is converted into electric energy, realize simultaneously the dirty water decontamination handles.
The cultivation of embodiment 1 gained MMR-1 bacterial strain
The MMR-1 bacterial strain is accessed in blue-green algae substratum BG11 take sodium acetate as carbon source with 1% inoculum size, and every liter of composition of BG11 substratum is as follows: 1500mg NaNO
3, 40mg K
2HPO
4, 75mgMgSO
4.7H
2O, 27mg CaCl
2, 1mg EDTA, 6mg citric acid, 6mg ferric ammonium citrate, 20mg Na
2CO
3, and 1ml trace element mixed liquor A 5(2860mg H
3BO
3, 1810mg MnCl
2.4H
2O, 222mg ZnSO
4.7H
2O, 390mg NaMoO
4.2H
2O, 79mg CuSO
4.5H
2O, 49mg Co(NO
3) 2.6H
2O).After incubated at room temperature 24 hours, obtain the biology pure growth of MMR-1 bacterial strain, be used for following microbiological fuel cell electrogenesis experiment.
MMR-1 bacterial strain also available other substratum is cultivated.But MMR-1 bacterial strain growing way on the blue-green algae substratum that adds carbon source or broth culture is better.
Contain the microbiological fuel cell electrogenesis experiment of embodiment 1 gained MMR-1 bacterial strain
Anolyte is the mixed solution of 195ml BG11 substratum and sodium acetate, and anode is carbon paper, and negative electrode is the air electrode of Pt/C catalysis, contains concentration 50mM phosphoric acid buffer in cathode compartment, with above-mentioned biology pure growth as the electrogenesis microbial inoculant in the anolyte compartment.In the microbiological fuel cell start-up course, the resistance of external circuit keeps 5000ohm; , the solution in microbiological fuel cell is Removed All during lower than 50mV when output voltage, be replaced by simultaneously new water inlet solution.Further, before experiment, experimental installation and electrode all carry out autoclaving, and the Bechtop that is seeded in of electrogenesis microorganism carries out, and avoid living contaminants.The microbiological fuel cell two ends adopt volt ohm-milliammeter every the 1min record once, and voltage and power stage are as shown in Figure 3.Under room temperature, through after the reaction of 100h, the microbiological fuel cell of the present embodiment begins that power stage is arranged, and its peak power can reach 56.3mW/m
2
The above the specific embodiment of the present invention does not consist of the restriction to protection domain of the present invention.Various other corresponding changes and distortion that any technical conceive according to the present invention has been done all should be included in the protection domain of claim of the present invention.
Claims (9)
1. a bacillus subtilis strain, is characterized in that, described bacterial strain is MMR-1, and preserving number is CGMCC No.6871.
2. bacillus subtilis strain claimed in claim 1, is characterized in that, the 16S rRNA gene order of described bacterial strain is SEQ ID No.1.
3. the biology pure growth of a bacterium strain, is characterized in that, having preserving number is all identification marks of the bacillus subtilis strain MMR-1 of CGMCC No.6871.
4. the application of the described biology pure growth of the described bacillus subtilis strain of claim 1 or claim 3 in electricity generation by microorganism.
5. a microbiological fuel cell, is characterized in that, the electrogenesis microorganism of described microbiological fuel cell is the described bacillus subtilis strain of claim 1 or the described biology pure growth of claim 3.
6. microbiological fuel cell claimed in claim 5, is characterized in that, the electron donor of described electrogenesis microorganism is the organism in glucose, sodium acetate, N.F,USP MANNITOL or sewage.
7. microbiological fuel cell claimed in claim 5, is characterized in that, comprises anolyte compartment, cathode compartment and external circuit three parts, and described anolyte compartment comprises anolyte, anode and electrogenesis microorganism.
8. microbiological fuel cell claimed in claim 7, is characterized in that, described anolyte is for adding blue-green algae substratum, broth culture or the sewage of carbon source.
9. microbiological fuel cell claimed in claim 5, is characterized in that, the negative electrode of described microbiological fuel cell is the air cathode of Pt/C catalysis.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105154366A (en) * | 2015-09-25 | 2015-12-16 | 哈尔滨理工大学 | Bacillus cereus capable of degrading phenol and having electrogenesis characteristic and application thereof |
| CN105304923A (en) * | 2015-09-25 | 2016-02-03 | 哈尔滨理工大学 | Method for increasing energy utilization rate of phenol-degradable microbial fuel cell |
| CN108285880A (en) * | 2018-01-04 | 2018-07-17 | 广州大学 | A kind of bacillus and its application with electricity production characteristic and denitrification activity |
| CN111320266A (en) * | 2020-02-28 | 2020-06-23 | 江苏大学 | Low-carbon-source-cost dye anaerobic biological decoloring system and method |
| CN111440740B (en) * | 2020-04-02 | 2021-01-12 | 盐城工学院 | Lactobacillus mucosae and application thereof in sewage desulfurization |
| CN117247147A (en) * | 2023-11-17 | 2023-12-19 | 山东吉昌龙环境工程有限公司 | Liquid composite carbon source for sewage treatment |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105154366A (en) * | 2015-09-25 | 2015-12-16 | 哈尔滨理工大学 | Bacillus cereus capable of degrading phenol and having electrogenesis characteristic and application thereof |
| CN105304923A (en) * | 2015-09-25 | 2016-02-03 | 哈尔滨理工大学 | Method for increasing energy utilization rate of phenol-degradable microbial fuel cell |
| CN105304923B (en) * | 2015-09-25 | 2017-12-08 | 哈尔滨理工大学 | A kind of method for the microbiological fuel cell capacity usage ratio for improving degradable phenol |
| CN105154366B (en) * | 2015-09-25 | 2018-07-06 | 哈尔滨理工大学 | One plant of degradable phenol and the bacillus cereus with electricity production characteristic |
| CN108285880A (en) * | 2018-01-04 | 2018-07-17 | 广州大学 | A kind of bacillus and its application with electricity production characteristic and denitrification activity |
| CN108285880B (en) * | 2018-01-04 | 2021-03-30 | 广州大学 | Bacillus with electrogenesis characteristic and denitrification activity and application thereof |
| CN111320266A (en) * | 2020-02-28 | 2020-06-23 | 江苏大学 | Low-carbon-source-cost dye anaerobic biological decoloring system and method |
| CN111440740B (en) * | 2020-04-02 | 2021-01-12 | 盐城工学院 | Lactobacillus mucosae and application thereof in sewage desulfurization |
| CN117247147A (en) * | 2023-11-17 | 2023-12-19 | 山东吉昌龙环境工程有限公司 | Liquid composite carbon source for sewage treatment |
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Application publication date: 20130605 |