CN110518273B - Single-cell microbial fuel cell for electrochemical active bacteriology research and preparation method thereof - Google Patents
Single-cell microbial fuel cell for electrochemical active bacteriology research and preparation method thereof Download PDFInfo
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- CN110518273B CN110518273B CN201910814486.5A CN201910814486A CN110518273B CN 110518273 B CN110518273 B CN 110518273B CN 201910814486 A CN201910814486 A CN 201910814486A CN 110518273 B CN110518273 B CN 110518273B
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01M—PROCESSES OR MEANS, e.g. BATTERIES, FOR THE DIRECT CONVERSION OF CHEMICAL ENERGY INTO ELECTRICAL ENERGY
- H01M4/00—Electrodes
- H01M4/86—Inert electrodes with catalytic activity, e.g. for fuel cells
- H01M4/88—Processes of manufacture
- H01M4/8878—Treatment steps after deposition of the catalytic active composition or after shaping of the electrode being free-standing body
- H01M4/8896—Pressing, rolling, calendering
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01M—PROCESSES OR MEANS, e.g. BATTERIES, FOR THE DIRECT CONVERSION OF CHEMICAL ENERGY INTO ELECTRICAL ENERGY
- H01M4/00—Electrodes
- H01M4/86—Inert electrodes with catalytic activity, e.g. for fuel cells
- H01M4/90—Selection of catalytic material
- H01M4/9075—Catalytic material supported on carriers, e.g. powder carriers
- H01M4/9083—Catalytic material supported on carriers, e.g. powder carriers on carbon or graphite
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01M—PROCESSES OR MEANS, e.g. BATTERIES, FOR THE DIRECT CONVERSION OF CHEMICAL ENERGY INTO ELECTRICAL ENERGY
- H01M4/00—Electrodes
- H01M4/86—Inert electrodes with catalytic activity, e.g. for fuel cells
- H01M4/96—Carbon-based electrodes
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01M—PROCESSES OR MEANS, e.g. BATTERIES, FOR THE DIRECT CONVERSION OF CHEMICAL ENERGY INTO ELECTRICAL ENERGY
- H01M8/00—Fuel cells; Manufacture thereof
- H01M8/16—Biochemical fuel cells, i.e. cells in which microorganisms function as catalysts
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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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- 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 a single-cell microbial fuel cell for electrochemical active bacteriology research and a preparation method thereof, belonging to the technical field of microbial electrochemistry. The invention aims to provide a simple and effective manufacturing method of a single-chamber air cathode microbial fuel cell for omics research, aiming at solving the problem that most microbial fuel cells are not suitable for omics research. The basic structure of the single-cell microbial fuel cell for electrochemical active bacteriology research is composed of a carbon paper anode, an air cathode and a glass bottle body. The invention adopts the rolling process to manufacture the air cathode, can simplify the operation flow and shorten the manufacturing time, improves the manufacturing efficiency of the air cathode, and ensures the pure culture environment of the reactor by arranging the sterilizing filter membrane at the outer side of the cathode. The carbon paper is used as the anode, so that the quantitative extraction of nucleic acid and protein samples can be ensured, and the subsequent high-throughput sequencing and omics analysis are facilitated.
Description
Technical Field
The invention belongs to the technical field of microbial electrochemistry; in particular to a preparation method of a single-chamber microbial fuel cell for electrochemical activity bacteriology research.
Background
In the field of sewage treatment, Microbial Fuel Cell (MFC) technology has been rapidly developed as a new wastewater treatment technology, and has become a research hotspot at home and abroad. In the last decade, with the optimization of reactor configuration and electrode materials, the electric energy output of the MFC system has been significantly improved. However, some studies found that MFC electrochemically active biofilm formation after amplification was difficult, easily leading to power overshoot, and reactor performance after amplification was far below laboratory level. Due to the lack of sufficient understanding of electrode microbial biofilm formation, it is difficult to achieve a simultaneous amplification of MFC power generation efficiency by macroscopic changes in electrode material and dimensions alone. The main contributors to electrocatalytic activity in MFCs are electrochemically active microorganisms concentrated at the anode. The electron transfer rate between the microorganism and the anode is a critical factor influencing the performance of the microbial electrochemical system, and becomes an important reason for restricting the electrocatalytic conversion efficiency of the microorganism. Therefore, in order to further improve the system efficiency, the optimization and control are performed at the initial stage of biofilm formation, and researches on the aspects of extracellular electron transfer mechanism of electrochemically active bacteria and communities, the optimization of the structure of the biofilm community on the surface of the anode, and the like need to be performed.
The single-chamber air cathode microbial fuel cell has good electricity generation effect and simple configuration, and becomes an effective tool for researching electrochemical bacterial function. Genomics, transcriptomics and proteomics based on high-throughput nucleic acid sequencing technology, which is emerging in recent years, provide ideas and methods for the research of characteristics and metabolic functions of functional microorganisms in different environments. However, most of the current single-chamber air cathode microbial fuel cells are not suitable for omics research, particularly the research of pure bacteria systems, due to the configuration and the electrode material. Therefore, in order to research the physiological and ecological characteristics and metabolic pathways of the electrochemically active bacteria by using a molecular biology technology of nucleic acid sequencing, the single-chamber air cathode microbial fuel cell must be further modified.
Disclosure of Invention
The invention aims to provide a simple and effective manufacturing method of a single-chamber air cathode microbial fuel cell for omics research, aiming at solving the problem that most of the conventional microbial fuel cells are not suitable for omics research.
In order to solve the problems, the single-cell microbial fuel cell for electrochemical activity bacteriology research provided by the invention comprises a reactor shell (1), a carbon paper anode (2), an air cathode (3), a sterilizing filter membrane (4), a lead (5), a resistor (6) and a titanium wire (7), wherein the shell of the reactor is a glass bottle body with an upper cover and a circular opening on the side wall, the air cathode is made of powdered capacitance activated carbon, conductive carbon black, stainless steel mesh and polytetrafluoroethylene emulsion, the carbon paper anode is arranged in the reactor shell and is connected with a titanium wire, the titanium wire passes through the upper cover of the reactor shell and extends out of the reactor shell, the resistor is connected with the other section of the titanium wire through a lead, and the other end of the titanium wire is connected with the air cathode; the air cathode is arranged at a circular opening on the side wall of the reactor shell, is fixed on the reactor shell through a single flange and a stainless steel clamp, completely seals the opening, and the outer side of the air cathode is provided with a sterilization filter membrane.
The size of the anode carbon paper is (2-2.5 cm) x (6-10 cm).
The effective volume of the glass bottle body is 200-250 ml, the opening of the glass bottle is sealed by a rubber plug to form an upper cover, and the circular opening area on one side of the bottle body is 6-8 cm2。
The aperture of the sterilizing filter membrane is 0.22 mu m, and the area of the sterilizing filter membrane is 8-10 cm2。
In addition, the invention also provides a preparation method of the single-cell microbial fuel cell for electrochemical activity bacteriology research, which comprises the following steps:
step one, mixing powdered capacitance activated carbon powder and absolute ethyl alcohol, then carrying out ultrasonic stirring, slowly adding polytetrafluoroethylene emulsion, continuously rolling by using a roller press until the mixture is pressed into a sheet after the absolute ethyl alcohol is evaporated to form a micelle, and preparing a catalyst layer of an air cathode;
mixing conductive carbon black and absolute ethyl alcohol, then carrying out ultrasonic stirring, slowly adding polytetrafluoroethylene emulsion, continuously rolling by using a roller press until the absolute ethyl alcohol is evaporated until the mixture forms a micelle, pressing into a sheet, then putting the sheet and a stainless steel mesh into the roller press for pressing so as to embed the sheet into the stainless steel mesh, and putting the mixed sheet into a muffle furnace for heating;
step three, superposing the catalyst layer prepared in the step one and the heated sheet prepared in the step two, putting the superposed sheets into a roller press, and pressing the superposed sheets for three to four times to obtain the air cathode;
cutting the carbon paper, perforating with titanium wires, and coating conductive adhesive for fixing to obtain a carbon paper anode;
and step five, assembling the air cathode, the sterilization filter membrane, the carbon paper anode and the glass bottle body into an integrated structure to obtain the single-chamber microbial fuel cell.
In the first step, the mass ratio of the capacitance activated carbon powder to the absolute ethyl alcohol is 1: (4-6), the ultrasonic stirring time is 20-40 min, and the thickness of the catalytic layer is 0.4-0.6 mm.
In the first step, the solid content of the polytetrafluoroethylene emulsion is 40-80 wt%, and the mass ratio of the capacitance activated carbon powder to the polytetrafluoroethylene emulsion is (4-6): 1.
In the second step, the mass ratio of the conductive carbon black to the absolute ethyl alcohol is 1: (2-3), the ultrasonic stirring time is 15-30 min, and the thickness of the slice is 0.4-0.6 mm.
In the second step, the solid content of the polytetrafluoroethylene emulsion is 40-80 wt%, and the mass ratio of the conductive carbon black to the polytetrafluoroethylene emulsion is (1-1.2): 3; the mesh number of the stainless steel mesh is 40-100 meshes.
Fifthly, assembling the air cathode, the sterilization filter membrane, the carbon paper anode and the glass bottle body into an integral structure, and assembling according to the following connection relation: the reactor shell is a glass bottle body with an upper cover and a circular opening on the side wall, the air cathode is made of powdered capacitance activated carbon, conductive carbon black, a stainless steel mesh and polytetrafluoroethylene emulsion, the carbon paper anode is arranged inside the reactor shell and connected with a titanium wire, the titanium wire penetrates through the upper cover of the reactor shell and extends out of the reactor shell, a resistor is connected through a lead, the resistor is connected with the other section of titanium wire through the lead, and the other end of the titanium wire is connected with the air cathode; the air cathode is arranged at a circular opening on the side wall of the reactor shell, is fixed on the reactor shell through a single flange and a stainless steel clamp, completely seals the opening, and the outer side of the air cathode is provided with a sterilization filter membrane.
Advantageous effects
The invention relates to a single-chamber microbial fuel cell for electrochemical active bacteriology research, the basic structure of which is composed of a carbon paper anode, an air cathode and a glass bottle body. The air cathode is made of powdered capacitance active carbon, conductive carbon black, stainless steel mesh and polytetrafluoroethylene emulsion, and the outer side of the air cathode is provided with a sterilization filter membrane. The activated carbon has stable chemical properties and good catalytic activity; the stainless steel mesh is a high-strength basic structure material and has good conductive performance, and the mesh structure of the stainless steel mesh can allow air to permeate through; the degerming filter membrane is arranged outside the cathode, so that the pure culture environment of the reactor can be ensured. The carbon paper is used as the anode, so that the quantitative extraction of nucleic acid and protein samples can be ensured, and the subsequent high-throughput sequencing and omics analysis are facilitated.
The single-chamber microbial fuel cell constructed by the invention is accessed with wild type and mutant strains under the condition of taking sodium acetate as a carbon source, and the stable output voltage period can reach more than 40 days. The anode DNA sample is extracted by the kit and then subjected to agarose gel electrophoresis, and the band is bright and clear, thereby achieving the library building standard of omics analysis.
Drawings
FIG. 1 is the voltage output of the single-chamber microbial fuel cell of the present invention when different strains are inoculated in pure culture.
FIG. 2 is an electrophoretogram of the anode DNA sample of the present invention after extraction.
FIG. 3 is a schematic diagram of the construction of a single-compartment microbial fuel cell reactor of the present invention; 1-reactor shell, 2-anode, 3-cathode, 4-sterilization filter membrane, 5-lead, 6-resistor and 7-titanium wire.
Detailed Description
Example 1 Single cell microbial Fuel cell construction for omics studies
The single-cell microbial fuel cell for omics research in the embodiment is an integrated structure consisting of a carbon paper anode, an air cathode and a glass bottle body, wherein the air cathode is made of powdered capacitance activated carbon, conductive carbon black, a stainless steel net and polytetrafluoroethylene emulsion, and a sterilization filter membrane is arranged on the outer side of the air cathode. The preparation method of the fuel cell is carried out according to the following steps:
firstly, mixing 10g of powdered capacitance activated carbon with 63ml of absolute ethyl alcohol for 30min under the condition of ultrasonic stirring, and slowly adding 1.333ml of polytetrafluoroethylene emulsion with the solid content of 60% in the process of ultrasonic stirring; and when the absolute ethyl alcohol is evaporated until the mixture forms a micelle, continuously rolling the mixture by using a roller press until the mixture is pressed into a sheet with the thickness of 0.5mm, thus obtaining the catalyst layer.
Secondly, mixing 6g of powdered carbon black with 120ml of absolute ethyl alcohol for 20min under the condition of ultrasonic stirring, and slowly adding 12ml of polytetrafluoroethylene emulsion with the solid content of 60 percent by weight in the process of ultrasonic stirring; when the absolute ethyl alcohol is evaporated until the mixture forms a micelle, continuously rolling the mixture by using a roller press until the mixture is pressed into a sheet with the thickness of 0.5mm, then putting the sheet and a stainless steel net with the mesh number of 80 meshes (the thickness is about 0.5mm) into the roller press to be pressed so as to be embedded into the stainless steel net, and putting the mixed sheet into a muffle furnace to heat for 20min at 350 ℃.
And thirdly, stacking the catalyst layer prepared in the step one and the mixed sheet into a roller press, and pressing for three to four times to obtain the 0.5mm air cathode.
Fourthly, cutting the carbon paper into strips of 2.5 multiplied by 8cm, perforating the carbon paper strips by using J-shaped titanium wires with the diameter of 0.5mm, and smearing conductive adhesive at the joints for fixing to obtain the carbon paper anode.
Fifthly, fixing the carbon paper anode into a reactor with the effective volume of 250mL, and enabling the titanium wire to extend out of the top of the reactor. The air cathode is arranged at one side of the reactor, the catalytic layer is arranged inwards, the outside of the diffusion layer is covered with a 0.22 mu m sterilization filter membrane, and the air cathode is fixed by using a single flange and a stainless steel clamp. And a section of titanium wire is used for leading out current at the cathode, and an external circuit connects the anode, the 1000 omega resistor and the cathode by a copper lead to complete the assembly of the single-chamber microbial fuel cell reactor.
The application comprises the following steps:
the single-chamber microbial fuel cell obtained in the embodiment is subjected to high-pressure steam sterilization on the whole (121 ℃, 20min), 5mL of bacterial liquid is inoculated, 1g/L of sodium acetate is used as a substrate, 100mM of phosphate buffer solution is used as a solvent, the single-chamber microbial fuel cell is operated in a 35 ℃ constant-temperature chamber in an intermittent non-continuous flow mode, wild type strains and mutant strains are inoculated under the condition that the sodium acetate is used as a carbon source, and the stable output voltage period can reach more than 40 days. The anode DNA sample is extracted by the kit and then subjected to agarose gel electrophoresis, and the band is bright and clear, thereby achieving the library building standard of omics analysis.
Claims (5)
1. A preparation method of a single-chamber microbial fuel cell for electrochemical activity bacteriology research is characterized in that: the method comprises the following steps:
step one, mixing powdered capacitance activated carbon powder and absolute ethyl alcohol, then carrying out ultrasonic stirring, slowly adding polytetrafluoroethylene emulsion, continuously rolling by using a roller press until the mixture is pressed into a sheet after the absolute ethyl alcohol is evaporated to form a micelle, and preparing a catalyst layer of an air cathode;
mixing conductive carbon black and absolute ethyl alcohol, then carrying out ultrasonic stirring, slowly adding polytetrafluoroethylene emulsion, continuously rolling by using a roller press until the absolute ethyl alcohol is evaporated until the mixture forms a micelle, pressing into a sheet, then putting the sheet and a stainless steel mesh into the roller press for pressing so as to embed the sheet into the stainless steel mesh, and putting the mixed sheet into a muffle furnace for heating;
step three, superposing the catalyst layer prepared in the step one and the heated sheet prepared in the step two, putting the superposed sheets into a roller press, and pressing the superposed sheets for three to four times to obtain the air cathode;
cutting the carbon paper, perforating with titanium wires, and coating conductive adhesive for fixing to obtain a carbon paper anode;
step five, assembling the air cathode, the sterilizing filter membrane, the carbon paper anode and the reactor into an integrated structure to prepare the single-chamber microbial fuel cell, which specifically comprises the following steps: the reactor shell is a glass bottle body with an upper cover and a circular opening on the side wall, the carbon paper anode is arranged inside the reactor shell and is connected with a titanium wire, the titanium wire penetrates through the upper cover of the reactor shell and extends out of the reactor shell, a resistor is connected through a lead, the resistor is connected with another section of titanium wire through a lead, and the other end of the other section of titanium wire is connected with the air cathode; the air cathode is arranged at the circular opening of the side wall of the reactor shell, is fixed on the reactor shell through a single flange and a stainless steel clamp, completely seals the opening, and the sterilizing filter membrane is arranged on the outer side of the air cathode.
2. The method for preparing a single cell microbial fuel cell for electrochemically active bacteriological studies according to claim 1, characterized in that: in the first step, the mass ratio of the capacitance activated carbon powder to the absolute ethyl alcohol is 1: (4-6), the ultrasonic stirring time is 20-40 min, and the thickness of the catalytic layer is 0.4-0.6 mm.
3. The method for preparing a single cell microbial fuel cell for electrochemically active bacteriological studies according to claim 1, characterized in that: in the first step, the solid content of the polytetrafluoroethylene emulsion is 40-80 wt%, and the mass ratio of the capacitance activated carbon powder to the polytetrafluoroethylene emulsion is (4-6): 1.
4. The method for preparing a single cell microbial fuel cell for electrochemically active bacteriological studies according to claim 1, characterized in that: in the second step, the mass ratio of the conductive carbon black to the absolute ethyl alcohol is 1: (2-3), the ultrasonic stirring time is 15-30 min, and the thickness of the slice is 0.4-0.6 mm.
5. The method for preparing a single cell microbial fuel cell for electrochemically active bacteriological studies according to claim 1, characterized in that: in the second step, the solid content of the polytetrafluoroethylene emulsion is 40-80 wt%, and the mass ratio of the conductive carbon black to the polytetrafluoroethylene emulsion is (1-1.2): 3; the mesh number of the stainless steel mesh is 40-100 meshes.
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