CN103627661B - Application of ochrobactrum in preparation of microbial fuel cell, and device and method for preparing microbial fuel cell - Google Patents

Application of ochrobactrum in preparation of microbial fuel cell, and device and method for preparing microbial fuel cell Download PDF

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CN103627661B
CN103627661B CN201310629966.7A CN201310629966A CN103627661B CN 103627661 B CN103627661 B CN 103627661B CN 201310629966 A CN201310629966 A CN 201310629966A CN 103627661 B CN103627661 B CN 103627661B
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fuel cell
microbiological fuel
anode
anthropi
ochrobactrum
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CN103627661A (en
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李鑫
乔琰
钟国祯
李长明
王行国
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Southwest University
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Southwest University
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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
    • 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
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P70/00—Climate change mitigation technologies in the production process for final industrial or consumer products
    • Y02P70/50—Manufacturing or production processes characterised by the final manufactured product

Abstract

The invention discloses application of ochrobactrum in preparation of a microbial fuel cell, and a device and a method for preparing the microbial fuel cell. The ochrobactrum (Ochrobactrum sp.5753rd-generation) is preserved by the China Center for Type Culture Collection (CCTCC), the preservation number is CCTCC NO:M2013549, and the preservation date is November 6, 2013. The ochrobactrum can transfer electrons to an extracellular electron acceptor under an anaerobic condition, and can utilize xylose as a single carbon source to generate electric energy; the ochrobactrum is strong in electricity production activity, and has a good application prospect in recycling utilization of a biological energy source.

Description

Anthropi is in the application prepared in microbiological fuel cell and apparatus and method thereof
Technical field
The invention belongs to biochemical fuel-cell field, particularly anthropi is preparing the application in microbiological fuel cell, also relates to the preparation method of microbiological fuel cell and the microbiological fuel cell prepared by anthropi.
Background technology
Along with the exhaustion day by day of the traditional fuels such as oil coal, find new green sustainable energy extremely urgent.Wood sugar is a kind of reproducible resource, comes from it and is extensively present in the agricultural wastes such as maize straw.If can effectively utilize wood sugar electrogenesis, not only contribute to the process of agricultural wastes, be also conducive to solving day by day serious energy problem.
Microbiological fuel cell (Microbial Fuel Cell, MFC) is by the metabolism oxidation of organic compounds of microorganism, and the electronics of generation, through a series of transmission, arrives the electrode outside born of the same parents, then is passed to negative electrode by external circuit; The proton produced arrives negative electrode by proton exchange membrane, then oxygen in negative electrode and electronics are combined and generate water, and therefore it is a kind of reproducible energy of cleaning, and MFC can organism in degradation water simultaneously, can be used for solution environmental problem.
Microorganism affects the very important factor of of microbiological fuel cell performance.All utilize mixed bacterium as catalyzer in the current microbiological fuel cell for decomposing wood sugar.Although the electrogenesis power of mixed bacterium fuel cell is higher, because the indefinite further raising hindering electricity generation ability of its electrogenesis bacterium and electrogenesis mechanism.The efficiency of fuel cell generation of single bacteria microorganism fuel cell is lower relative to mixed bacterium fuel cell, but electrogenesis mechanism is clear and definite, and this electrogenesis principle contributing to being familiar with further microbiological fuel cell is to continue its discharge capability of raising.
Therefore, be badly in need of single bacteria microorganism that a kind of efficiency of fuel cell generation is high, electrogenesis principle and continuation for microorganisms fuel cell improve microorganism discharge capability and lay the foundation.
Summary of the invention
In view of this, an object of the present invention is to provide anthropi Ochrobactrum sp.5753 rd-generation; Two of object of the present invention is to provide anthropi preparing the application in microbiological fuel cell; Three of object of the present invention is to provide the microbiological fuel cell utilizing anthropi to prepare; Four of object of the present invention is the preparation method providing microbiological fuel cell.
For achieving the above object, following technical scheme is provided:
1. anthropi Ochrobactrum sp.5753 rd-generation, by China typical culture collection center preservation, deposit number is CCTCC NO:M2013549.
2. anthropi described in is preparing the application in microbiological fuel cell.
3. utilize microbiological fuel cell prepared by described anthropi.
Preferably, described microbiological fuel cell comprises anode cavities and cathode cavity, separate by proton exchange membrane between described anode cavities and cathode cavity, described anode cavities and cathode cavity are respectively arranged with anode and negative electrode, described anode is connected by wire with negative electrode, described anode cavities is the nutrient solution containing anthropi, and described nutrient solution take wood sugar as carbon source, and described cathode cavity is the phosphate buffered saline buffer of the six directions Tripotassium iron hexacyanide.
In the present invention, proton exchange membrane does following process before using, and proton exchange membrane is boiled 20 minutes with sulfuric acid, deionized water that hydrogen peroxide, deionized water, massfraction that massfraction is 0.5% are 1% successively.
Preferably, described electrode is carbon brush or carbon-based electrode.
Described wire is plain conductor, is preferably titanium silk.
Preferably, the concentration of the phosphate buffered saline buffer of the described six directions Tripotassium iron hexacyanide is 20-200mmol, and optimum concn is 50mmol.
Preferred, in described nutrient solution, each concentration of component is as follows: sodium bicarbonate 10gL -1, SODIUM PHOSPHATE, MONOBASIC 8.5gL -1, yeast extract 5gL -1, wood sugar 10gL -1.
4. the preparation method of microbiological fuel cell described in, comprises the steps:
(1) anode cavities and cathode cavity proton exchange membrane are separated, then by wire, anode is connected with negative electrode;
(2) it is in the substratum of carbon source with wood sugar that the anthropi of taking the logarithm vegetative period adds fresh, then anode cavities is added, and in anode chamber, pass into nitrogen removal oxygen, then in cathode cavity, add the phosphate buffered saline buffer of the six directions Tripotassium iron hexacyanide, obtain microbiological fuel cell.
Preferably, described proton exchange membrane does following process before using, and proton exchange membrane is boiled 20 minutes with sulfuric acid, deionized water that hydrogen peroxide, deionized water, massfraction that massfraction is 0.5% are 1% successively.
Beneficial effect of the present invention is: the invention discloses anthropi Ochrobactrum sp.5753 rd-generation, this bacterium has comparatively forceful electric power chemically reactive, can under anaerobic transmit electronics to extracellular electron acceptor, as the catalyst oxidation wood sugar of MFC, can produce electric energy while oxidation wood sugar; Utilizing the MFC of strain construction disclosed by the invention to produce maximum output voltage is 0.552V, and peak power output density is 2625mWm -3, this is than large about 20 times of the mixed bacterium MFC of decomposition wood sugar of the same type.Meanwhile, this bacterial strain is amphimicrobian type, in the process of electrogenesis, do not need strict anaerobic environment, this reduces the requirement to device in practical application.
Accompanying drawing explanation
In order to make object of the present invention, technical scheme and beneficial effect clearly, the invention provides following accompanying drawing:
Fig. 1 is the output voltage figure of three domestication bacterium MFC.
Fig. 2 is the output power density figure of three domestication bacterium MFC.
Fig. 3 remains wood sugar figure in the anolyte of three domestication bacterium MFC.
Fig. 4 is the cyclic voltammogram of three domestication bacterium bacteria suspensions.
Culture presevation
Anthropi Ochrobactrum sp.5753 in the present invention rd-generation obtains efficiency of fuel cell generation high bacterial strain through domestication from anthropi (Ochrobactrum) sp.575 containing the substratum of wood sugar, send China typical culture collection center preservation, deposit number is CCTCC NO:M2013549, address is positioned at Wuhan, China Wuhan University, preservation date is on November 6th, 2013, and Classification And Nomenclature is Ochrobactrum sp.5753 rd-generation.
Embodiment
Below in conjunction with specific embodiment, the present invention is more specifically described in detail, but embodiments of the present invention are not limited thereto, to the processing parameter do not indicated especially, can refer to routine techniques and carry out.
The substratum used in the present invention is as follows:
The each component concentration of xylose media is as follows: sodium bicarbonate 10gL -1, SODIUM PHOSPHATE, MONOBASIC 8.5gL -1, yeast extract 5gL -1, wood sugar 10gL -1.
In LB substratum, each component concentration is as follows: peptone 10gL -1, yeast extract 5gL -1, sodium-chlor 10gL -1, agar 16gL -1.
Embodiment 1
Get the anthropi Ochrobactrum sp.575 of Laboratories Accession, streak culture rear picking list colony inoculation is in xylose media, and under temperature is 37 DEG C of conditions, Anaerobic culturel 10 is little of OD 600=0.8-1.0, then by bacterium liquid at 6000r/min, within centrifugal 5 minutes, collect thalline, then rejoin fresh xylose media for 4 DEG C, bacteria suspension is made in mixing, and makes bacteria suspension OD 600be 1.2.
Start microbiological fuel cell (Microbial Fuel Cell, MFC), the microbiological fuel cell that the present embodiment uses is two cylindrical ponds (single pond useful volume 100mL), middle alternating floor proton exchange membrane, utilize clip to be connected together in two ponds, then use rubber stopper seal; Carbon brush, as anode and negative electrode, is that wire does electronics collector with titanium silk; The bacteria suspension of preparation is joined the anode cavities of microbiological fuel cell, then the oxygen in logical nitrogen 30 minutes removing bacteria suspensions; During startup, the phosphate buffered saline buffer of 100mL containing the 50mmol six directions Tripotassium iron hexacyanide is added in cathode cavity, extrernal resistance is 1800 Ω, run under temperature is 18 ~ 25 DEG C of conditions, and start time open circuit voltage, record maximum output voltage (output voltage (U) of MFC gathers by digital multimeter) in operational process, result is as shown in table 1.
After this MFC discharges for some time, get microbionation and cultivate in solid LB media from anode cavities, cultivate after one day, observe the form of bacterium colony, the characteristics such as color, single bacterium that picking surface characteristic difference and bacterium colony are before as broad as long is as once taming bacterium.Then get and once tame bacterium repetition above-mentioned steps 2 times, obtain three domestication bacterium, and record its open circuit voltage and maximum output voltage, the results are shown in Table 1.
The electric discharge of table 1. original bacteria and domestication bacterium is compared
Bacterial strain Open circuit voltage (mV) Maximum output voltage (mV)
Original anthropi 180 124
Once tame bacterium 330 192
Secondary admonitory talk to subordinates bacterium 383 226
Three domestication bacterium 680 552
As shown in Table 1, the electrogenesis effect of original anthropi is also not obvious, and after three domestications, the electricity generation ability of training bacterium improves a lot.Then send China typical culture collection center preservation by three domestication bacterium, deposit number is CCTCCNO:M2013549, called after Ochrobactrum sp.5753 rd-generation.
Embodiment 2
Get single colony inoculation in xylose media after three domestication anthropi embodiment 1 obtained are streak culture, under temperature is 37 DEG C of conditions, Anaerobic culturel 10 is little of OD 600=0.8-1.0, then by bacterium liquid under 6000r/min condition 4 DEG C centrifugal 5 minutes, collect thalline, will add fresh xylose media in thalline, bacteria suspension is made in mixing, and controls bacteria suspension OD 600be 1.2.
Start microbiological fuel cell, microbiological fuel cell is two cylindrical ponds (single pond useful volume 100mL), and middle alternating floor proton exchange membrane, utilizes clip to be connected together in two ponds, then use rubber stopper seal; Carbon brush, as anode and negative electrode, does electronics collector with titanium silk wire.Above-mentioned bacteria suspension is joined the anode cavities of microbiological fuel cell, then the oxygen in logical nitrogen 30 minutes removing bacteria suspensions.During startup, add the phosphate buffered saline buffer of 100mL containing the 50mmol six directions Tripotassium iron hexacyanide in cathode cavity, extrernal resistance is 1800 Ω, and temperature is run under 18 ~ 25 conditions.
The output voltage (U) of MFC gathers by digital multimeter, the experimental result of the output voltage of MFC as shown in Figure 1, as shown in Figure 1, in MFC after inoculation 0-20 hour that the anthropi after three domestications builds, voltage rises fast, reaches maximum value at 24 hours output voltages.After each replaced medium, voltage reaches maximum value all rapidly, and runs 4 stable repeatably output voltages of all after dates appearance, and maximum output voltage reaches 0.552V.
Electric current (I) is calculated by Ohm's law I=U/R and obtains, and R is external loop resistance, and the drafting of polarization curve and power density curve is by regulating external loop resistance, and after measuring corresponding output voltage, calculate and curve plotting, result as shown in Figure 2.Current density I sformula I is utilized respectively with volumetric power density P s=U/(RV) and P=U 2/ (RV) calculates, and wherein V is the useful volume of anode cavities.As shown in Figure 2, when extrernal resistance is 325 Ω, acquisition maximum power density is 2625mWm -3.
From MFC after electric discharge, every 4 hours bacteria suspensions that take a morsel, leave supernatant liquor after centrifugal, utilize the residual content of wood sugar in the supernatant of measurement of ultraviolet-visible spectrophotometer bacteria suspension, result as shown in Figure 3.Result shows, and in 24 hours that cultivate, wood sugar is consumed fast, and after 20 hours, the wear rate of wood sugar slows down, and it is 0.2mg/mL that a final discharge cycle terminates remaining Xylose Content in rear anolyte.
The present embodiment shows, the running status of three training bacterium in MFC is good, can wood sugar be utilized to produce higher electric energy under not adding exogenous electron to transmit the condition of intermediate.
Embodiment 3
Get single colony inoculation in xylose media after three domestication anthropi embodiment 1 obtained are streak culture, under temperature is 37 DEG C of conditions, Anaerobic culturel 10 is little of OD 600=0.8-1.0, then gets bacterium liquid under 6000r/min, 4 DEG C of conditions centrifugal 5 minutes, collects thalline, then thalline is added fresh xylose media, and bacteria suspension is made in mixing, and controls bacteria suspension OD 600be 1.2.Add not containing in the substratum of wood sugar after simultaneously thalline is centrifugal by identical method, obtained bacteria suspension, in contrast, and with the fresh xylose media not inoculating thalline for blank.
Above-mentioned bacteria suspension or xylose media are added in electrolyzer respectively, utilize electrochemical workstation (CHI660E, Shanghai occasion China) constant potential (0.2V) is carried out to bacterium liquid simulate electric discharge 12 hours, and then carry out the cyclic voltammetry of three-electrode system, using carbon brush as working electrode with to electrode (being greater than working electrode to electrode surface area), Ag/AgCl electrode is reference electrode, sweeps speed for 30mVs -1, sweep limit is-0.6V ~ 0V, and its result as shown in Figure 4.As shown in Figure 4, two pairs of redox peaks have been there are at the cyclic voltammogram of the xylose media having bacterium, and at aseptic xylose media with have without wood sugar in the cyclic voltammogram of the substratum of bacterium and do not occur redox peak, illustrate that bacterial strain is be carbon source electrogenesis with wood sugar and create redox materials really.
The explanation of this example, bacterial strain can utilize the mode in the middle of bacterial strain self metabolic wood sugar secretion electron transmission to transmit electronics under the condition of not adding exogenous electron intermediate, there is stronger electrochemical activity, can reduce this bacterium bioenergy reclaim in practical application cost, for its bioenergy reclaim in application provide guarantee.
What finally illustrate is, above preferred embodiment is only in order to illustrate technical scheme of the present invention and unrestricted, although by above preferred embodiment to invention has been detailed description, but those skilled in the art are to be understood that, various change can be made to it in the form and details, and not depart from claims of the present invention limited range.

Claims (10)

1. anthropi ( ochrobactrumsp.) 575 3 rd-generation, by China typical culture collection center preservation, deposit number is CCTCC NO:M 2013549.
2. anthropi described in claim 1 is preparing the application in microbiological fuel cell.
3. utilize microbiological fuel cell prepared by anthropi described in claim 1.
4. microbiological fuel cell according to claim 3, it is characterized in that: comprise anode cavities and cathode cavity, separate by proton exchange membrane between described anode cavities and cathode cavity, described anode cavities and cathode cavity are respectively arranged with anode and negative electrode, described anode is connected by wire with negative electrode, described anode cavities is the nutrient solution containing anthropi, and described nutrient solution take wood sugar as carbon source, and described cathode cavity is the phosphate buffered saline buffer of the six directions Tripotassium iron hexacyanide.
5. microbiological fuel cell according to claim 4, is characterized in that: described anode electrode or cathode electrode are carbon brush.
6. microbiological fuel cell according to claim 4, is characterized in that: described wire is titanium silk.
7. microbiological fuel cell according to claim 4, is characterized in that: the concentration of the phosphate buffered saline buffer of the described six directions Tripotassium iron hexacyanide is 20-200mmol.
8. the microbiological fuel cell according to any one of claim 4-7, is characterized in that: in described nutrient solution, each concentration of component is as follows: sodium bicarbonate 10gL -1, SODIUM PHOSPHATE, MONOBASIC 8.5gL -1, yeast extract 5gL -1, wood sugar 10gL -1.
9. the preparation method of microbiological fuel cell described in any one of claim 4-8, is characterized in that, comprise the steps:
(1) anode cavities and cathode cavity proton exchange membrane are separated, then by wire, anode is connected with negative electrode;
(2) it is in the substratum of carbon source with wood sugar that the anthropi of taking the logarithm vegetative period adds fresh, then anode cavities is added, and in anode chamber, pass into nitrogen removal oxygen, then in cathode cavity, add the phosphate buffered saline buffer of the six directions Tripotassium iron hexacyanide, obtain microbiological fuel cell.
10. preparation method according to claim 9, it is characterized in that, described proton exchange membrane does following process before using, and proton exchange membrane is boiled 20 minutes with sulfuric acid, deionized water that hydrogen peroxide, deionized water, massfraction that massfraction is 0.5% are 1% successively.
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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103153883A (en) * 2011-05-04 2013-06-12 Rm泰科株式会社 Method for heavy metal elimination or precious metal recovery using microbial fuel cell

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103153883A (en) * 2011-05-04 2013-06-12 Rm泰科株式会社 Method for heavy metal elimination or precious metal recovery using microbial fuel cell

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
微生物染料电池的功能拓展和机理解析;孙敏;《中国博士学位论文全文数据库》;20111115(第11期);全文 *
微生物燃料电池中阳极产电微生物的研究进展;范平 等;《生物学通报》;20111231;第46卷(第10期);6－9 *
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