CN107285477B - Preparation for accelerating anode biofilm formation efficiency of gram-negative electrogenic bacteria in microbial fuel cell and application - Google Patents
Preparation for accelerating anode biofilm formation efficiency of gram-negative electrogenic bacteria in microbial fuel cell and application Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/005—Combined electrochemical biological processes
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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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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2305/00—Use of specific compounds during water treatment
- C02F2305/06—Nutrients for stimulating the growth of microorganisms
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- 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
Abstract
The invention discloses a preparation for accelerating the anode biofilm formation efficiency of gram-negative electrogenic bacteria in a microbial fuel cell and application thereof, wherein the preparation is prepared from the following components in percentage by mass, 10-60% of signal molecules; 10% -60% of ammonium salt; 10% -20% of signal molecule protective agent; 10 to 20 percent of sustained release agent. And (3) feeding the wastewater to be treated into an anode chamber of the microbial fuel cell, and synchronously or sequentially adding the preparation for accelerating the anode film formation efficiency of the gram-negative electricity-producing bacteria in the microbial fuel cell and inoculating the gram-negative electricity-producing bacteria into the wastewater in any order. The preparation provided by the invention can improve the growth speed of the biological membrane, shorten the membrane hanging time of the biological membrane, has the characteristics of low cost and wide applicability, and lays a foundation for the expanded engineering application of the microbial fuel cell.
Description
Technical Field
The invention relates to the technical field of sewage treatment, in particular to a preparation for accelerating the anode biofilm formation efficiency of gram-negative electrogenic bacteria in a microbial fuel cell.
Background
Energy shortage and environmental pollution are two major problems faced by China nowadays. In recent centuries, with the rapid development of industrial revolution, energy has become the fundamental guarantee and development power for human survival. But the energy crisis is gradually highlighted due to the rapid consumption and population proliferation of fossil energy (coal, oil, natural gas, etc.). In addition, the environment of the water area of China is seriously polluted by a large amount of waster water which is discharged in a stealing mode or is discharged out of the standard. The total amount of wastewater discharged in China from 2013 to 2015 reaches 2400 hundred million tons. In 2015, the discharge amount of Chemical Oxygen Demand (COD) in the wastewater is 2300 million tons, the operation cost of treatment facilities is up to 1100 million yuan, and the direct energy consumption is higher than 100 million kilowatt hours. Under the stress of huge energy crisis, the consumption of a large amount of energy to eliminate organic pollutants in wastewater is really an unaddressed way, so that the development of new energy and renewable energy sources is urgently needed to relieve the pressure of energy demand and treat the organic pollutants in wastewater. The organic pollutants in the wastewater are actually low-grade chemical energy, and the development of Microbial Fuel Cells (MFCs) provides an effective way for recycling the low-grade chemical energy. The self-renewable electrogenesis microorganism is used as a catalyst, and the organic matters in the wastewater are used as electron donors to realize electrogenesis at normal temperature and normal pressure. However, at present, the MFCs are far from reaching the practical electrification application level under the limitation of factors such as engineering electrode materials and the like.
The anode material acts as a biological carrier and electron acceptor for MFCs and is the primary limiting component for current transport. Carbon materials with various configurations, such as carbon plates, carbon cloth, carbon felt and the like, have excellent biocompatibility and specific surface area, and are the most common anode materials in the research of MFCs at present. However, the carbon material has large resistance and the conductivity is only 3x104～1x105Sm-1At least three orders of magnitude lower than metal electrodes, severely limiting their development in engineering applications. The metal material has beneficial ductility and conductivity, and is a cheap and scalable engineering electrode material. Some leather orchards at presentAlthough the negative bacteria have good electricity generating performance, the biological membrane of the negative bacteria is difficult to adsorb or embed on a metal electrode carrier, so that the MFCs are difficult to start or even cannot be started, the electricity generating performance and the pollutant treatment performance of the bacteria are influenced, and the engineering expanded application of the bacteria is influenced.
Quorum Sensing (QS) is a mechanism of information transmission among bacteria, i.e., bacteria synthesize and secrete self-induced molecules (AI) as signal molecules, and detect the concentration of the signal molecules, and when the concentration of the AI reaches a certain threshold along with the population density of the bacteria, specific gene expression is initiated.
The Chinese patent application with the application number of 201611068535.8 discloses a device and a method for improving the biofilm formation efficiency of a wastewater treatment biofilm. Although the film hanging efficiency is high, the device is complex, the cost is high and the universal adaptation is difficult.
Based on the preparation, the invention provides a preparation for accelerating the anode biofilm formation efficiency of gram-negative electrogenic bacteria in a microbial fuel cell. The preparation has the advantages of high film forming efficiency, simple preparation, relatively low cost and universal applicability.
Disclosure of Invention
Aiming at the problems that some gram-negative bacteria have good electrogenesis performance in the prior art, but the metal electrode has low film forming efficiency, high cost and low applicability, the invention provides a preparation and a method for accelerating the film forming efficiency of the gram-negative electrogenesis bacteria anode in a microbial fuel cell.
A preparation for accelerating the anode film forming efficiency of gram-negative electrogenic bacteria in a microbial fuel cell comprises the following components in percentage by mass:
the sum of the mass percentages of the components is 100 percent.
Further preferably, the composition is prepared from the following components in percentage by mass:
signal molecule: 30 to 50 percent
Signal molecule protective agent: 10 to 15 percent of
Ammonium salt: 30 to 50 percent
Sustained release agent: 10 to 15 percent of
The sum of the mass percentages of the components is 100 percent.
Still more preferably, the composition is prepared from the following components in percentage by mass:
signal molecule: 30 to 50 percent
Signal molecule protective agent: 10 percent of
Ammonium salt: 30 to 50 percent
Sustained release agent: 10 percent.
The invention also discloses a method for reducing natural degradation of signal molecules by adding dimethyl sulfoxide, wherein a slow release agent is prepared into a mother solution, and the mother solution is coated by a spraying technology to reduce the action time of the preparation in waste water, and the durability of the effect of the preparation is maintained by double protection of the signal molecule protective agent and the slow release agent.
Quorum Sensing (QS) is a mechanism of information transfer between bacteria cells, i.e., bacteria synthesize and secrete an auto-inducer (AI) as a signal molecule, and detect the concentration of the signal molecule, and when the AI concentration reaches a certain threshold value along with the population density of the bacteria, specific gene expression is initiated. QS is currently known to regulate a variety of important functions of bacteria such as biofilm formation and the like. Bacteria produce biofilms that enhance resistance to external disturbances. At present, the reactor using the metal material as the anode in the microbial fuel cell is difficult to start up because the microorganism is difficult to form the biofilm on the metal carrier.
The preparation for promoting anode biofilm formation is prepared by mixing different types of signal molecules and ammonium salt, can accelerate the formation of the metal anode biofilm and enable the metal anode biofilm to have good development on a metal carrier, has the remarkable advantages of fast start and long and stable period, and is an economic and efficient preparation.
The signal molecules are AH L s, which comprise N-butyroyl-homeoserine lacton (C4-HS L), N-hexanooyl-homeoserine lacton (C6-HS L0), N-octanooyl-homeoserine (C8-HS L), N-decanoyl-D L2-homeoserine (C10-HS L), N-laurooyl-D L-homeoserine (C12-HS L), N-3-oxo-hexanooyl-homeoserine (3-oxo-C6-HS L), N-3-oxo-octanoyl-homeoserine (3-oxo-C8-HS L), N- (3-oxo-decanooyl) -L-oxorosclerol (3-oxo-C63593), and the above mentioned organic solvent from Aldolone-HS 593-24, Sigma-3-aldeoxide-HS 599.
Preferably, the signal molecule is formed by mixing the following components in percentage by mass:
C4-HSL：5-50％
C6-HSL：5-50％
C8-HSL：5-50％
C10-HSL：5-50％
C12-HSL：5-50％
3-oxo-C6-HSL：5-50％
3-oxo-C8-HSL：5-50％
3-oxo-C10-HSL：5-50％
3-oxo-C12-HSL：5-50％
pC-HSL:5-50％。
the formation, development and function regulation of most gram-negative electrogenic microbial biofilms require quorum sensing signal molecules to participate in the information transfer between bacteria in bacterial biofilms, AH L is a proven intercellular information transfer molecule in bacterial biofilms under natural or culture conditions, AH L plays a key role in forming and maintaining the three-dimensional structure of bacterial biofilms.
The sum of the mass percentages of the components is 100 percent.
Further preferably, the signal molecule consists of the following components in percentage by mass:
C4-HSL：5～20％
C6-HSL：5～20％
C8-HSL：5～20％
C10-HSL：5-20％
C12-HSL：5-20％
3-oxo-C6-HSL：5-20％
3-oxo-C8-HSL：5-20％
3-oxo-C10-HSL：5-20％
3-oxo-C12-HSL：5～20％
pC-HSL：5～20％
the sum of the mass percentages of the components is 100 percent.
Still further preferably, the signal molecule consists of, in mass percent:
C4-HSL：5～15％
C6-HSL：5～15％
C8-HSL：5～15％
C10-HSL：5-15％
C12-HSL：5-15％
3-oxo-C6-HSL：5-15％
3-oxo-C8-HSL：5-15％
3-oxo-C10-HSL：5-15％
3-oxo-C12-HSL：5～15％
pC-HSL：5～15％。
more preferably, the signal molecules comprise 8-12% by mass of C4-HS L%
C6-HSL：8～12％
C8-HSL：8～12％
C10-HSL：8～12％
C12-HSL：8～12％
3-oxo-C6-HSL：8～12％
3-oxo-C8-HSL：8～12％
3-oxo-C10-HSL：8～12％
3-oxo-C12-HSL：8～12％
pC-HSL：8～12％。
Most preferably, the signal molecule consists of, in mass percent:
C4-HSL：10％
C6-HSL：10％
C8-HSL：10％
C10-HSL：10％
C12-HSL：10％
3-oxo-C6-HSL：10％
3-oxo-C8-HSL：10％
3-oxo-C10-HSL：10％
3-oxo-C12-HSL：10％
pC-HSL：10％。
preferably, the signal molecule protector is dimethyl sulfoxide (DMSO).
Preferably, the ammonium salt is at least one of ammonium chloride, ammonium sulfate, ammonium nitrate and ammonium carbonate.
Preferably, the sustained release agent comprises the following components in percentage by mass:
ethyl cellulose: 40 to 60 percent
Diethyl phthalate: 10 to 30 percent
PEG6000:10％-30％。
The sum of the mass percentages of the components is 100 percent.
Further preferably, the sustained release agent is present in the formulation in the form of a coating film.
In order to control the release rate of the preparation in a reactor and achieve better film forming efficiency, a sustained release agent needs to be added. The coating film coated on the sustained or controlled release agent prepared by the coating technique is not a single, pure entity. In order to form a coating film with certain permeability and mechanical properties, the coating material must be prepared into a coating solution by using an optimal coating formula, and a certain process is adopted for coating to form a coating film with slow and controlled release effects.
The preparation of the invention is prepared by the following method:
weighing all the components required by various signal molecules according to the mass percent of the components, dissolving and mixing the components uniformly by using ethyl acetate, blowing off the components by using nitrogen, adding ammonium salt weighed according to the mass percent of the components and a signal molecule protective agent, stirring the mixture uniformly, pressing the mixture into a sheet preparation, and coating the coating film outside the preparation by using a rolling spray coating technology. And (5) obtaining a finished product after the inspection is qualified.
The preparation prepared by the invention needs to be added with a slow release material in order to control the release rate of the preparation in a reactor and enable the preparation to stably and continuously exert the effect, the invention adopts a rolling spray coating technology, and the preparation can play a slow release role, and because the film forming agent and most of auxiliary additives are high molecular materials with excellent physical and chemical properties, the coated film can be moisture-proof, light-proof, wear-resistant and the like, meanwhile, the operation is easy to master, the homogeneity is good, the drying is fast, the quality is easy to preserve when the preparation is used for signal molecules which are easy to damage at high temperature, and the excellent performance of the preparation is achieved.
The invention also provides an application of the preparation in wastewater treatment of a microbial fuel cell.
Preferably, the dosage of the preparation in the microbial fuel cell is 50-200 mg/L.
The preparation prepared by the invention is applied to a microbial fuel cell, wherein the anode electrogenesis microorganisms are gram negative electrogenesis bacteria such as Pseudomonas aeruginosa (Pseudomonas aeruginosa), thioredoxin (Geobacter sulfuriduiduccus) and the like. The metal electrode material of the anode adopts a titanium sheet of 1cm by 2cm or a stainless steel felt of 1cm by 2 cm. And (4) comparing the addition of the preparation and the non-addition of the preparation in the reactor, and comparing the generated biofilm amount, the biofilm thickness and the biofilm formation capacity.
The invention aims to realize engineering expansion application, but the main factor influencing the engineering expansion application at present is the defect of an engineering electrode, namely the lack of a high-quality anode. The electrode commonly used in the laboratory is mainly made of carbon materials, but the electrode is high in resistance and low in conductivity and is not suitable for development of engineering application, and the metal material has excellent ductility and conductivity and can be very suitable for engineering electrode materials. However, the metal material itself has poor biocompatibility, which limits the development of engineering application. Based on the method, the stainless steel felt and the titanium sheet are used as metal anode materials, and a foundation is laid for realizing engineering application.
A method for accelerating the anode film forming efficiency of gram-negative electrogenic bacteria in a microbial fuel cell comprises the following steps:
the wastewater to be treated is sent into an anode chamber of a microbial fuel cell, and the preparation and the gram-negative electricity-generating bacteria are synchronously or sequentially added into the wastewater in any order.
Compared with the prior art, the method has the following beneficial effects:
the preparation has good economic and environmental benefits, and has the advantages of improving the growth speed of the biological membrane, shortening the membrane hanging time of the biological membrane, reducing the working dose, reducing the use cost, being simple to operate and having wide practicability. The preliminary detection and judgment are carried out by adopting the preliminary bench tests, so that the risk born in the engineering implementation process can be reduced to the greatest extent, and powerful guarantee and guidance are provided for the expanded application of the reactor.
Drawings
FIG. 1 is a flow chart of sample injection and reaction biofilm formation.
FIG. 2 is a flow chart of the preparation of the formulation of the present invention.
Detailed Description
The following examples are provided to further illustrate the present invention, but are not intended to limit the scope of the present invention.
Example 1:
(1) the preparation method comprises the steps of weighing all components required by various signal molecules according to the mass percentage (C4-HS L10%, C6-HS L10%, C8-HS L10%, C10-HS L10%, C12-HS L10%, 3-oxo-6-HS L10%, 3-oxo-C8-HS L10%, 3-oxo-C10-HS L10%, 3-oxo-C12-HS L10%, and pC-HS L10%), dissolving and mixing uniformly by using ethyl acetate, then blowing off by using nitrogen, adding ammonium chloride and dimethyl sulfoxide weighed according to the mass percentage, and stirring uniformly (signal molecules 50%, ammonium chloride 30%, and dimethyl sulfoxide 10%, pressing the signal molecules, ammonium chloride and dimethyl sulfoxide into a tablet, wherein the tablet specification is that the diameter is 2cm, the thickness is 5mm, preparing a coating liquid, and weighing the components according to the mass percentage, namely 60% of ethyl cellulose, 30% of diethyl phthalate, 010% of outer coating film, and finally preparing a finished product by using a rolling coating film-making technology (such as a finished product is shown in a drawing after inspection flow chart 2).
(2) Constructing a microbial fuel cell: 50ml of simulated wastewater is put into an anode chamber of a microbial fuel cell, the microbial fuel cell device adopts an H-shaped double-chamber structure and consists of two glass bottles, a cation exchange membrane is adopted in the middle, the two chambers are fastened and connected by a clamp, a titanium sheet electrode is adopted as an anode electrode, a graphite electrode is adopted as a cathode electrode, a lead is connected by a titanium wire, a connecting part is stuck by conductive gel, and an external circuit is connected with a fixed resistor of 1 ohm.
(3) Media and test microorganisms:
the culture medium is a simulated wastewater culture medium (NaAc 1 g/L; NaCl 0.55 g/L; MgSO 2)40.1g/L； K2HPO43.4g/L；KH2PO44.4g/L；NaHCO32g/L；NH4Cl 0.1g/L)
Microorganisms: gram-negative bacterium, Pseudomonas aeruginosa (Pseudomonas aeruginosa)
(3) Control group: experimental setup controls, one set: adding sodium acetate into an M9 culture medium and adding a preparation to culture the tested microorganism; the other group: culturing the tested microorganism by using M9 culture medium and sodium acetate
(4) The operation conditions are as follows: mixing the tested microorganism bacterial solutions with late growth log phase obtained from the culture medium, transferring 10ml, inoculating into the anode of the microorganism fuel cell, adjusting pH of the whole device to 7.0-7.5 with sodium acetate as electron donor and potassium ferricyanide as electron acceptor, placing in a 37 deg.C constant temperature incubator, and operating
(5) Sample adding conditions: the preparation prepared by the invention enters the reactor in an adding mode. The feeding position is a sample feeding position at the peripheral opening of the H-shaped reactor, the sample feeding frequency is once in five days, and one preparation is fed once. The flow of sample introduction and reaction biofilm formation is shown in figure 1.
(6) Detection indexes of the biological membrane are as follows: the amount of the biofilm was characterized by measuring protease by Coomassie Brilliant blue method, the thickness of the biofilm by laser confocal method and the biofilm formation efficiency by reactor start-up time, the results are shown in Table 1
Example 2:
(1) the preparation method comprises the steps of weighing all components required by various signal molecules according to the mass percentage (C4-HS L10%, C6-HS L10%, C8-HS L10%, C10-HS L10%, C12-HS L10%, 3-oxo-6-HS L10%, 3-oxo-C8-HS L10%, 3-oxo-C10-HS L10%, 3-oxo-C12-HS L10%, and pC-HS L10%), dissolving and mixing uniformly by using ethyl acetate, then blowing off by using nitrogen, adding ammonium chloride and dimethyl sulfoxide weighed according to the mass percentage, stirring uniformly (signal molecules 50%, ammonium chloride 30%, dimethyl sulfoxide 10%, coating liquid 10%), pressing the signal molecules, ammonium chloride and dimethyl sulfoxide into a tablet, preparing the coating liquid with the diameter of 2cm and the thickness of 5mm, weighing ethyl cellulose outer coating 60%, diethyl phthalate 30%, and PEG 600010% according to the mass percentage, and spraying to obtain a finished product after the finished product is qualified by using a coating film-rolling technology.
(2) Constructing a microbial fuel cell: 50ml of simulated wastewater is put into an anode chamber of a microbial fuel cell, the microbial fuel cell device adopts an H-shaped double-chamber structure and consists of two glass bottles, a cation exchange membrane is adopted in the middle, the two chambers are fastened and connected by a clamp, a titanium sheet electrode is adopted as an anode electrode, a graphite electrode is adopted as a cathode electrode, a lead is connected by a titanium wire, a connecting part is stuck by conductive gel, and an external circuit is connected with a fixed resistor of 1 ohm.
(3) Media and test microorganisms:
the culture medium is a simulated wastewater culture medium (NaAc 1 g/L; NaCl 0.55 g/L; MgSO 2)40.1g/L； K2HPO43.4g/L；KH2PO44.4g/L；NaHCO32g/L；NH4Cl 0.1g/L)
Microorganisms: gram-negative bacteria-Thiodegen bacillus (Geobacter sulfuridunens)
(3) Control group: experimental setup controls, one set: adding sodium acetate into an M9 culture medium and adding a preparation to culture the tested microorganism; the other group: culturing the tested microorganism by using M9 culture medium and sodium acetate
(4) The operation conditions are as follows: mixing the tested microorganism bacterial solutions with late growth log phase obtained from the culture medium, transferring 10ml, inoculating into the anode of the microorganism fuel cell, adjusting pH of the whole device to 7.0-7.5 with sodium acetate as electron donor and potassium ferricyanide as electron acceptor, placing in a 37 deg.C constant temperature incubator, and operating
(5) Sample adding conditions: the preparation prepared by the invention enters the reactor in an adding mode. The feeding position is a sample feeding position at the peripheral opening of the H-shaped reactor, the sample feeding frequency is once in five days, and one preparation is fed once
(6) Detection indexes of the biological membrane are as follows: the amount of the biofilm was characterized by measuring protease by Coomassie Brilliant blue method, the thickness of the biofilm by laser confocal method and the biofilm formation efficiency by reactor start-up time, the results are shown in Table 1
Example 3:
(1) the preparation method comprises the steps of weighing all components required by various signal molecules according to the mass percentage (C4-HS L10%, C6-HS L10%, C8-HS L10%, C10-HS L10%, C12-HS L10%, 3-oxo-6-HS L10%, 3-oxo-C8-HS L10%, 3-oxo-C10-HS L10%, 3-oxo-C12-HS L10%, and pC-HS L10%), dissolving and mixing uniformly by using ethyl acetate, then blowing off by using nitrogen, adding ammonium chloride and dimethyl sulfoxide weighed according to the mass percentage, stirring uniformly (30% of the signal molecules, 50% of the ammonium chloride, 10% of the dimethyl sulfoxide, and 10% of coating liquid), pressing the signal molecules, the ammonium chloride and the dimethyl sulfoxide into a tablet, preparing the coating liquid with the specification of 2cm diameter and 5mm thickness, weighing 60% of ethyl cellulose, 30% of diethyl phthalate and 30% of polyethylene glycol (PEG) according to the mass percentage, and spraying and checking the coating liquid to obtain the finished product after rolling and checking the qualified coating film preparation by using a coating technology.
(2) Constructing a microbial fuel cell: 50ml of simulated wastewater is put into an anode chamber of a microbial fuel cell, the microbial fuel cell device adopts an H-shaped double-chamber structure and consists of two glass bottles, a cation exchange membrane is adopted in the middle, the two chambers are fastened and connected by a clamp, a titanium sheet electrode is adopted as an anode electrode, a graphite electrode is adopted as a cathode electrode, a lead is connected by a titanium wire, a connecting part is stuck by conductive gel, and an external circuit is connected with a fixed resistor of 1 ohm.
(3) Media and test microorganisms:
the culture medium is a simulated wastewater culture medium (NaAc 1 g/L; NaCl 0.55 g/L; MgSO 2)40.1g/L； K2HPO43.4g/L；KH2PO44.4g/L；NaHCO32g/L；NH4Cl 0.1g/L)
Microorganisms: gram-negative bacterium, Pseudomonas aeruginosa (Pseudomonas aeruginosa)
(3) Control group: experimental setup controls, one set: adding sodium acetate into an M9 culture medium and adding a preparation to culture the tested microorganism; the other group: culturing the tested microorganism by using M9 culture medium and sodium acetate
(4) The operation conditions are as follows: mixing the tested microorganism bacterial solutions with late growth log phase obtained from the culture medium, transferring 10ml, inoculating into the anode of the microorganism fuel cell, adjusting pH of the whole device to 7.0-7.5 with sodium acetate as electron donor and potassium ferricyanide as electron acceptor, placing in a 37 deg.C constant temperature incubator, and operating
(5) Sample adding conditions: the preparation prepared by the invention enters the reactor in an adding mode. The feeding position is a sample feeding position at the peripheral opening of the H-shaped reactor, the sample feeding frequency is once in five days, and one preparation is fed once
(6) Detection indexes of the biological membrane are as follows: the amount of the biofilm was characterized by measuring protease by Coomassie Brilliant blue method, the thickness of the biofilm by laser confocal method and the biofilm formation efficiency by reactor start-up time, the results are shown in Table 1
Example 4:
(1) the preparation method comprises the steps of weighing all components required by various signal molecules according to the mass percentage (C4-HS L10%, C6-HS L10%, C8-HS L10%, C10-HS L10%, C12-HS L10%, 3-oxo-6-HS L10%, 3-oxo-C8-HS L10%, 3-oxo-C10-HS L10%, 3-oxo-C12-HS L10%, and pC-HS L10%), dissolving and mixing uniformly by using ethyl acetate, then blowing off by using nitrogen, adding ammonium chloride and dimethyl sulfoxide weighed according to the mass percentage, stirring uniformly (30% of the signal molecules, 50% of the ammonium chloride, 10% of the dimethyl sulfoxide, and 10% of coating liquid), pressing the signal molecules, the ammonium chloride and the dimethyl sulfoxide into a tablet, preparing the coating liquid with the specification of 2cm diameter and 5mm thickness, weighing 60% of ethyl cellulose, 30% of diethyl phthalate and 30% of polyethylene glycol (PEG) according to the mass percentage, and spraying and checking the coating liquid to obtain the finished product after rolling and checking the qualified coating film preparation by using a coating technology.
(2) Constructing a microbial fuel cell: 50ml of simulated wastewater is put into an anode chamber of a microbial fuel cell, the microbial fuel cell device adopts an H-shaped double-chamber structure and consists of two glass bottles, a cation exchange membrane is adopted in the middle, the two chambers are fastened and connected by a clamp, a titanium sheet electrode is adopted as an anode electrode, a graphite electrode is adopted as a cathode electrode, a lead is connected by a titanium wire, a connecting part is stuck by conductive gel, and an external circuit is connected with a fixed resistor of 1 ohm.
(3) Media and test microorganisms:
the culture medium is a simulated wastewater culture medium (NaAc 1 g/L; NaCl 0.55 g/L; MgSO 2)40.1g/L；K2HPO43.4g/L；KH2PO44.4g/L；NaHCO32g/L；NH4Cl 0.1g/L)
Microorganisms: gram-negative bacteria-Thiodegen bacillus (Geobacter sulfuridunens)
(3) Control group: experimental setup controls, one set: adding sodium acetate into an M9 culture medium and adding a preparation to culture the tested microorganism; the other group: culturing the tested microorganism by using M9 culture medium and sodium acetate
(4) The operation conditions are as follows: mixing the tested microorganism bacterial solutions with late growth log phase obtained from the culture medium, transferring 10ml, inoculating into the anode of the microorganism fuel cell, adjusting pH of the whole device to 7.0-7.5 with sodium acetate as electron donor and potassium ferricyanide as electron acceptor, placing in a 37 deg.C constant temperature incubator, and operating
(5) Sample adding conditions: the preparation prepared by the invention enters the reactor in an adding mode. The feeding position is a sample feeding position at the peripheral opening of the H-shaped reactor, the sample feeding frequency is once in five days, and one preparation is fed once
(6) Detection indexes of the biological membrane are as follows: the amount of the biofilm was characterized by measuring protease by Coomassie Brilliant blue method, the thickness of the biofilm by laser confocal method and the biofilm formation efficiency by reactor start-up time, the results are shown in Table 1
Example 5:
(1) the preparation method comprises the steps of weighing all components required by various signal molecules according to the mass percentage (C4-HS L10%, C6-HS L10%, C8-HS L10%, C10-HS L10%, C12-HS L10%, 3-oxo-6-HS L10%, 3-oxo-C8-HS L10%, 3-oxo-C10-HS L10%, 3-oxo-C12-HS L10%, and pC-HS L10%), dissolving and mixing uniformly by using ethyl acetate, then blowing off by using nitrogen, adding ammonium chloride and dimethyl sulfoxide weighed according to the mass percentage, stirring uniformly (signal molecules 50%, ammonium chloride 30%, dimethyl sulfoxide 10%, coating liquid 10%), pressing the signal molecules, ammonium chloride and dimethyl sulfoxide into a tablet, preparing the coating liquid with the diameter of 2cm and the thickness of 5mm, weighing ethyl cellulose outer coating 60%, diethyl phthalate 30%, and PEG 600010% according to the mass percentage, and spraying to obtain a finished product after the finished product is qualified by using a coating film-rolling technology.
(2) Constructing a microbial fuel cell: 50ml of simulated wastewater is put into an anode chamber of a microbial fuel cell, the microbial fuel cell device adopts an H-shaped double-chamber structure and consists of two glass bottles, a cation exchange membrane is adopted in the middle, the two chambers are fastened and connected by a clamp, an anode electrode adopts a stainless steel felt electrode, a cathode electrode adopts a graphite electrode, a lead is connected by a titanium wire, a connecting part is stuck by conductive gel, and an external circuit is connected with a fixed resistor of 1 ohm.
(3) Media and test microorganisms:
the culture medium is a simulated wastewater culture medium (NaAc 1 g/L; NaCl 0.55 g/L; MgSO 2)40.1g/L； K2HPO43.4g/L；KH2PO44.4g/L；NaHCO32g/L；NH4Cl 0.1g/L)
Microorganisms: gram-negative bacterium, Pseudomonas aeruginosa (Pseudomonas aeruginosa)
(3) Control group: experimental setup controls, one set: adding sodium acetate into an M9 culture medium and adding a preparation to culture the tested microorganism; the other group: culturing the tested microorganism by using M9 culture medium and sodium acetate
(4) The operation conditions are as follows: mixing the tested microorganism bacterial solutions with late growth log phase obtained from the culture medium, transferring 10ml, inoculating into the anode of the microorganism fuel cell, adjusting pH of the whole device to 7.0-7.5 with sodium acetate as electron donor and potassium ferricyanide as electron acceptor, placing in a 37 deg.C constant temperature incubator, and operating
(5) Sample adding conditions: the preparation prepared by the invention enters the reactor in an adding mode. The feeding position is a sample feeding position at the peripheral opening of the H-shaped reactor, the sample feeding frequency is once in five days, and one preparation is fed once
(6) Detection indexes of the biological membrane are as follows: the amount of the biofilm was characterized by measuring protease by Coomassie Brilliant blue method, the thickness of the biofilm by laser confocal method and the biofilm formation efficiency by reactor start-up time, the results are shown in Table 1
Example 6:
(1) the preparation method comprises the steps of weighing all components required by various signal molecules according to the mass percentage (C4-HS L10%, C6-HS L10%, C8-HS L10%, C10-HS L10%, C12-HS L10%, 3-oxo-6-HS L10%, 3-oxo-C8-HS L10%, 3-oxo-C10-HS L10%, 3-oxo-C12-HS L10%, and pC-HS L10%), dissolving and mixing uniformly by using ethyl acetate, then blowing off by using nitrogen, adding ammonium chloride and dimethyl sulfoxide weighed according to the mass percentage, stirring uniformly (signal molecules 50%, ammonium chloride 30%, dimethyl sulfoxide 10%, coating liquid 10%), pressing the signal molecules, ammonium chloride and dimethyl sulfoxide into a tablet, preparing the coating liquid with the specification of 2cm diameter and 5mm thickness, weighing ethyl cellulose 60% according to the mass percentage, coating diethyl phthalate 30%, and PEG 600010% to obtain a finished product, and spraying and checking that the finished product is qualified by using a coating film-making rolling coating technology.
(2) Constructing a microbial fuel cell: 50ml of simulated wastewater is put into an anode chamber of a microbial fuel cell, the microbial fuel cell device adopts an H-shaped double-chamber structure and consists of two glass bottles, a cation exchange membrane is adopted in the middle, the two chambers are fastened and connected by a clamp, an anode electrode adopts a stainless steel felt electrode, a cathode electrode adopts a graphite electrode, a lead is connected by a titanium wire, a connecting part is stuck by conductive gel, and an external circuit is connected with a fixed resistor of 1 ohm.
(3) Media and test microorganisms:
the culture medium is a simulated wastewater culture medium (NaAc 1 g/L; NaCl 0.55 g/L; MgSO 2)40.1g/L； K2HPO43.4g/L；KH2PO44.4g/L；NaHCO32g/L；NH4Cl 0.1g/L)
Microorganisms: gram-negative bacteria-Thiodegen bacillus (Geobacter sulfuridunens)
(3) Control group: experimental setup controls, one set: adding sodium acetate into an M9 culture medium and adding a preparation to culture the tested microorganism; the other group: culturing the tested microorganism by using M9 culture medium and sodium acetate
(4) The operation conditions are as follows: mixing the tested microorganism bacterial solutions with late growth log phase obtained from the culture medium, transferring 10ml, inoculating into the anode of the microorganism fuel cell, adjusting pH of the whole device to 7.0-7.5 with sodium acetate as electron donor and potassium ferricyanide as electron acceptor, placing in a 37 deg.C constant temperature incubator, and operating
(5) Sample adding conditions: the preparation prepared by the invention enters the reactor in an adding mode. The feeding position is a sample feeding position at the peripheral opening of the H-shaped reactor, the sample feeding frequency is once in five days, and one preparation is fed once
(6) Detection indexes of the biological membrane are as follows: the amount of the biofilm was characterized by measuring protease by Coomassie Brilliant blue method, the thickness of the biofilm by laser confocal method and the biofilm formation efficiency by reactor start-up time, the results are shown in Table 1
Example 7:
(1) the preparation method comprises the steps of weighing all components required by various signal molecules according to the mass percentage (C4-HS L10%, C6-HS L10%, C8-HS L10%, C10-HS L10%, C12-HS L10%, 3-oxo-6-HS L10%, 3-oxo-C8-HS L10%, 3-oxo-C10-HS L10%, 3-oxo-C12-HS L10%, and pC-HS L10%), dissolving and mixing uniformly by using ethyl acetate, then blowing off by using nitrogen, adding ammonium chloride and dimethyl sulfoxide weighed according to the mass percentage, stirring uniformly (30% of the signal molecules, 50% of the ammonium chloride, 10% of the dimethyl sulfoxide, and 10% of coating liquid), pressing the signal molecules, the ammonium chloride and the dimethyl sulfoxide into a tablet, preparing the coating liquid with the specification of 2cm diameter and 5mm thickness, weighing 60% of ethyl cellulose, 30% of diethyl phthalate and 30% of polyethylene glycol (PEG) according to the mass percentage, and spraying and checking the coating liquid to obtain the finished product after rolling and checking the qualified coating film preparation by using a coating technology.
(2) Constructing a microbial fuel cell: 50ml of simulated wastewater is put into an anode chamber of a microbial fuel cell, the microbial fuel cell device adopts an H-shaped double-chamber structure and consists of two glass bottles, a cation exchange membrane is adopted in the middle, the two chambers are fastened and connected by a clamp, an anode electrode adopts a stainless steel felt electrode, a cathode electrode adopts a graphite electrode, a lead is connected by a titanium wire, a connecting part is stuck by conductive gel, and an external circuit is connected with a fixed resistor of 1 ohm.
(3) Media and test microorganisms:
the culture medium is a simulated wastewater culture medium (NaAc 1 g/L; NaCl 0.55 g/L; MgSO 2)40.1g/L； K2HPO43.4g/L；KH2PO44.4g/L；NaHCO32g/L；NH4Cl 0.1g/L)
Microorganisms: gram-negative bacterium, Pseudomonas aeruginosa (Pseudomonas aeruginosa)
(3) Control group: experimental setup controls, one set: adding sodium acetate into an M9 culture medium and adding a preparation to culture the tested microorganism; the other group: culturing the tested microorganism by using M9 culture medium and sodium acetate
(4) The operation conditions are as follows: mixing the tested microorganism bacterial solutions with late growth log phase obtained from the culture medium, transferring 10ml, inoculating into the anode of the microorganism fuel cell, adjusting pH of the whole device to 7.0-7.5 with sodium acetate as electron donor and potassium ferricyanide as electron acceptor, placing in a 37 deg.C constant temperature incubator, and operating
(5) Sample adding conditions: the preparation prepared by the invention enters the reactor in an adding mode. The feeding position is a sample feeding position at the peripheral opening of the H-shaped reactor, the sample feeding frequency is once in five days, and one preparation is fed once
(6) Detection indexes of the biological membrane are as follows: the amount of the biofilm was characterized by measuring protease by Coomassie Brilliant blue method, the thickness of the biofilm by laser confocal method and the biofilm formation efficiency by reactor start-up time, the results are shown in Table 1
Example 8:
(1) the preparation method comprises the steps of weighing all components required by various signal molecules according to the mass percentage (C4-HS L10%, C6-HS L10%, C8-HS L10%, C10-HS L10%, C12-HS L10%, 3-oxo-6-HS L10%, 3-oxo-C8-HS L10%, 3-oxo-C10-HS L10%, 3-oxo-C12-HS L10%, and pC-HS L10%), dissolving and mixing uniformly by using ethyl acetate, then blowing off by using nitrogen, adding ammonium chloride and dimethyl sulfoxide weighed according to the mass percentage, stirring uniformly (30% of the signal molecules, 50% of the ammonium chloride, 10% of the dimethyl sulfoxide, and 10% of coating liquid), pressing the signal molecules, the ammonium chloride and the dimethyl sulfoxide into a tablet, preparing the coating liquid with the specification of 2cm diameter and 5mm thickness, weighing 60% of ethyl cellulose, 30% of diethyl phthalate and 30% of polyethylene glycol (PEG) according to the mass percentage, and spraying and checking the coating liquid to obtain the finished product after rolling and checking the qualified coating film preparation by using a coating technology.
(2) Constructing a microbial fuel cell: 50ml of simulated wastewater is put into an anode chamber of a microbial fuel cell, the microbial fuel cell device adopts an H-shaped double-chamber structure and consists of two glass bottles, a cation exchange membrane is adopted in the middle, the two chambers are fastened and connected by a clamp, an anode electrode adopts a stainless steel felt electrode, a cathode electrode adopts a graphite electrode, a lead is connected by a titanium wire, a connecting part is stuck by conductive gel, and an external circuit is connected with a fixed resistor of 1 ohm.
(3) Media and test microorganisms:
the culture medium is a simulated wastewater culture medium (NaAc 1 g/L; NaCl 0.55 g/L; MgSO 2)40.1g/L； K2HPO43.4g/L；KH2PO44.4g/L；NaHCO32g/L；NH4Cl 0.1g/L)
Microorganisms: gram-negative bacteria-Thiodegen bacillus (Geobacter sulfuridunens)
(3) Control group: experimental setup controls, one set: adding sodium acetate into an M9 culture medium and adding a preparation to culture the tested microorganism; the other group: culturing the tested microorganism by using M9 culture medium and sodium acetate
(4) The operation conditions are as follows: mixing the tested microorganism bacterial solutions with late growth log phase obtained from the culture medium, transferring 10ml, inoculating into the anode of the microorganism fuel cell, adjusting pH of the whole device to 7.0-7.5 with sodium acetate as electron donor and potassium ferricyanide as electron acceptor, placing in a 37 deg.C constant temperature incubator, and operating
(5) Sample adding conditions: the preparation prepared by the invention enters the reactor in an adding mode. The feeding position is a sample feeding position at the peripheral opening of the H-shaped reactor, the sample feeding frequency is once in five days, and one preparation is fed once
(6) Detection indexes of the biological membrane are as follows: the amount of the biofilm was characterized by measuring protease by Coomassie Brilliant blue method, the thickness of the biofilm by laser confocal method and the biofilm formation efficiency by reactor start-up time, the results are shown in Table 1
TABLE 1
The above description is only an embodiment of the present invention, but the technical features of the present invention are not limited thereto, and any person skilled in the relevant art can change or modify the present invention within the scope of the present invention.
Claims (9)
1. A preparation for accelerating the anode biofilm formation efficiency of gram-negative electrogenic bacteria in a microbial fuel cell is characterized by comprising the following components in percentage by mass:
2. the preparation for accelerating the anode biofilm formation efficiency of gram-negative electrogenic bacteria in a microbial fuel cell according to claim 1, wherein the signal molecules are formed by mixing the following components in percentage by mass:
C4-HSL：5-50％
C6-HSL：5-50％
C8-HSL：5-50％
C10-HSL：5-50％
C12-HSL：5-50％
3-oxo-C6-HSL：5-50％
3-oxo-C8-HSL：5-50％
3-oxo-C10-HSL：5-50％
3-oxo-C12-HSL：5-50％
pC-HSL:5-50％。
3. the preparation for accelerating the anode biofilm culturing efficiency of gram-negative electrogenic bacteria in a microbial fuel cell according to claim 1, wherein the signal molecule protective agent is dimethyl sulfoxide.
4. The formulation for accelerating the anodic biofilm formation efficiency of gram-negative electrogenic bacteria in a microbial fuel cell of claim 1, wherein said ammonium salt is at least one of ammonium chloride, ammonium sulfate, ammonium nitrate and ammonium carbonate.
5. The preparation for accelerating the anode biofilm formation efficiency of gram-negative electrogenic bacteria in a microbial fuel cell according to claim 1, wherein the slow release agent comprises the following components in percentage by mass:
ethyl cellulose: 40 to 60 percent
Diethyl phthalate: 10 to 30 percent
PEG6000: 10％-30％。
6. The preparation for accelerating the anode biofilm culturing efficiency of gram-negative electrogenic bacteria in a microbial fuel cell according to claim 5, wherein the slow-release agent is present in the preparation in the form of a coating film.
7. Use of the agent for accelerating the anodic biofilm formation efficiency of gram-negative electrogenic bacteria in a microbial fuel cell according to claim 1 in the treatment of wastewater by a microbial fuel cell.
8. A method for accelerating the anode biofilm formation efficiency of gram-negative electrogenic bacteria in a microbial fuel cell is characterized by comprising the following steps:
feeding wastewater to be treated into an anode chamber of a microbial fuel cell, synchronously or sequentially adding a preparation for accelerating the anode film forming efficiency of gram-negative electricity-producing bacteria in the microbial fuel cell according to any one of claims 1 to 6 into the wastewater and inoculating the gram-negative electricity-producing bacteria in the microbial fuel cell in any sequence;
the microbial fuel cell takes a metal material as an anode.
9. The method of claim 8, wherein the formulation is added to the microbial fuel cell in an amount of 50-200 mg/L.
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