CN109022495B - Method for producing methane by reducing carbon dioxide with microorganisms - Google Patents
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Abstract
The invention relates to a method for producing methane by reducing carbon dioxide with microorganisms, which comprises the following steps: 1) constructing a microbial electrolysis cell comprising a cathode cell and an anode cell, wherein the cathode cell comprises a working electrode and a reference electrode; the anode cell includes a counter electrode; the cathode pool and the anode pool are separated by a proton exchange membrane; 2) adding methanogen culture medium into the cathode pool and the anode pool; 3) inoculating 1.5-5.0% methanogen bacteria liquid into the cathode pool; culturing at room temperature; 4) the cathode cell was charged with-0.6 v (vs SHE) voltage; 5) methane was harvested from the headspace of the cathode cell. Compared with the prior art, the invention does not need chemical catalyst, and has the advantages of cleanness, high efficiency and low cost2The method for biologically reducing the methane has no other by-products and has wide application prospect.
Description
Technical Field
The invention relates to CO2The field of resource utilization, in particular to a method for promoting microorganisms to utilize electric energy to convert CO2A process for the conversion to methane.
Background
Since the industrial revolution, the production activities of human beings have significantly increased atmospheric CO2The content of (a). CO 22As one of the major greenhouse gases, the increase in their content in the atmosphere may lead to an increase in global warming and climatic extremes. How to reduce CO in the atmosphere2The conversion of the content of (a) into even high value-added chemicals is a hot spot of global research. Common transformation methods include chemical transformation and biological transformation. Compared with the chemical method which usually needs high temperature and high pressure and a catalyst, the biological conversion can be carried out under the conditions of room temperature and normal pressure, the reaction condition is mild, and the method has wide application prospect.
Using microorganisms to convert CO2The conversion to methane has been extensively studied and reported. Patent No. CN 102925492A inoculates sludge-enriched microorganisms in the cathode compartment and reduces CO by means of electric current2Can produce acetic acid while producing methane. But the potential of the methane production (from minus 850mV to minus 950mV, vs Ag/AgCl) is higher, and the energy consumption is larger; and acetic acid as a byproduct is generated in the process of producing methane, so that the efficiency of producing methane is reduced, and the difficulty of product separation is increased.
In this regard, we provide a new biocathode capable of sequestering CO at lower potentials2Efficient low-cost conversion to CH4. The working electrode is made by utilizing the high specific surface area and porosity of the nitric acid-acetic acid mixed fiber film and the excellent conductivity of the carbon nano tube. The methanogen sarcina can be attached to the working electrode when placed in a cathode pool, and can efficiently and quickly attach CO when potential is applied to the cathode2Is converted into methane.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a method for producing methane by reducing carbon dioxide with microorganisms.
The purpose of the invention can be realized by the following technical scheme: a method for producing methane by reducing carbon dioxide with microorganisms is characterized by comprising the following steps:
1) constructing a microbial electrolytic cell comprising a cathode cell and an anode cell, wherein the cathode cell comprises electrolyte, a working electrode (cathode), a reference electrode and methanogen; the anode pool comprises electrolyte and a platinum counter electrode; the cathode pool and the anode pool are separated by an N117 proton exchange membrane;
2) adding methanogen culture medium into the cathode pool and the anode pool;
3) 3ml of methanogen bacteria liquid is inoculated in the cathode pool and cultured;
4) the cathode cell was charged with-0.6 v (vs SHE) voltage;
5) methane was harvested from the headspace of the cathode cell.
The working electrode in the step (1) is made of a nitric acid-acetic acid mixed fiber film adhered with the multi-wall carbon nano tube. The amount of the multi-walled carbon nano-tubes adhered to the nitric acid-acetic acid mixed fiber film is 0.1-10 mg/cm2The length of the multi-wall carbon nanotube is 0.5-50 μm; the aperture of the nitric acid-acetic acid mixed fiber film is 0.2-2 mu m.
The preparation method of the working electrode comprises the following steps: firstly, a multi-walled carbon nanotube with the length of 0.5-50 mu m is soaked in 5% hydrogen peroxide and vibrated for 1h, the multi-walled carbon nanotube is dissolved in 0.5-20% nafion ethanol solution after being dried, the multi-walled carbon nanotube is uniformly dispersed by ultrasonic treatment for 1-60 min to prepare the multi-walled carbon nanotube nafion ethanol solution, and then the multi-walled carbon nanotube nafion ethanol solution is uniformly coated on a nitric acid-acetic acid mixed fiber film with the aperture of 0.2-2 mu m and dried in the environment of 20-65 ℃. The loading capacity of the multi-walled carbon nano-tube of the nitric acid-acetic acid mixed fiber film is 0.1-10 mg/cm2. The nitric acid-acetic acid mixed fiber membrane loaded with the multi-walled carbon nanotubes was placed in a cathode cell and linked with a platinum wire to form a working electrode.
The cell body, the working electrode, the reference electrode, the counter electrode and the like are assembled into an electrolytic cell. 50mL of a common methanogen culture medium is introduced into both the cathode pool and the anode pool of the electrolytic cell.
Culturing the methanogen of step (2)The formula of the base is as follows: 0.35g/L K2HPO4，0.23g/L KH2PO4，0.5g/L NH4Cl，0.5g/L MgSO4·7H2O，0.25g/L CaCl2，2.25g/L NaCl，0.85g/L NaHCO3，0.5g/L Na2S·H2O, 10ml/L of trace element solution and 1ml/L of vitamin solution; the head space of the electrolytic cell is CO2:N21:4 (n/n);
the formula of the trace element solution is as follows: 1500mg/L FeCl2·4H2O，70mg/L ZnCl2，100mg/L MnCl2·4H2O，6mg/L H3BO3，190mg/L CoCl2·6H2O，2mg/L CuCl2·H2O，24mg/L NiCl2·6H2O，36mg/L NaMo4·2H2O，HCl(25％)10ml/L；
The formula of the vitamin solution is as follows: 2mg/L biotin, 2mg/L folic acid, 10mg/L vitamin B65mg/L vitamin B15mg/L vitamin B25mg/L nicotinic acid, 5mg/L pantothenic acid, 0.1mg/L vitamin B125mg/L aminobenzoic acid and 5mg/L lipoic acid.
The methanogen in step (3) includes but is not limited to methanogen pasteurella (Methanosarcina barkeri), methanogen brucellosis (Methanobacterium bryranti) and methanogen henneri (methanopirilum hungathei).
Culturing the methanogen in the step (3) at room temperature.
And (3) inoculating 1.5-5.0% methanogen bacteria liquid into the cathode pool. In order to avoid other products, the bacterial liquid should consist of a single methanogen. The reaction was initiated by applying a potential of-0.6V (vs SHE) to the cathode and the methane content of the headspace gas was determined by gas chromatography.
Compared with the prior art, the working electrode consists of a nitric acid-acetic acid mixed fiber film loaded with the multi-wall carbon nano tube, and the fiber film is horizontally placed at the bottom of the cathode pool. Meanwhile, methanogens are inoculated in the cathode pool, and CO can be separated by microorganisms under lower negative potential2Reduction to methane. The invention provides a needn't changeChemical catalyst, clean, high-efficiency and low-cost CO synthesis2The method for biologically reducing the methane has no other by-products and has wide application prospect.
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FIG. 1 shows a structural water body of the microbial electrolysis cell of the invention
Detailed Description
The invention is described in detail below with reference to the figures and specific embodiments.
Example 1:
this example was carried out simultaneously in experimental group a and control groups B, C:
the experimental group A is shown in FIG. 1 (examples two and three). The microbial electrolysis cell comprises a cathode cell and an anode cell, wherein a reference electrode 2, a cathode 4, methanogen 3 and a culture medium 5 are arranged in the cathode cell, a counter electrode 7 is arranged in the anode cell, a proton exchange membrane 6 is arranged between the cathode cell and the anode cell, and a power supply 1 is further arranged and is respectively connected with the reference electrode 2, the cathode 4 and the counter electrode 7. In this example, methanogen 3 is methanogen pasteurella (methanosarina barkeri). The reference electrode 2 is an Ag/AgCl electrode, and the cathode 4 is made of a nitric acid-acetic acid mixed fiber film adhered to a multi-walled carbon nanotube. The counter electrode 7 is a platinum counter electrode.
The formulation of medium 5 was as follows: 0.35g/L K2HPO4，0.23g/L KH2PO4，0.5g/L NH4Cl，0.5g/L MgSO4·7H2O，0.25g/L CaCl2，2.25g/L NaCl，0.85g/L NaHCO3，0.5g/L Na2S·H2O, 10ml/L of trace element solution and 1ml/L of vitamin solution; the head space of the electrolytic cell is CO2:N21:4 (n/n); the formula of the trace element solution is as follows: 1500mg/L FeCl2·4H2O，70mg/L ZnCl2，100mg/L MnCl2·4H2O，6mg/L H3BO3，190mg/L CoCl2·6H2O，2mg/L CuCl2·H2O，24mg/L NiCl2·6H2O，NiCl2mg/L NaMo4·2H2O, HCl (25%) 10 ml/L; the formula of the vitamin solution is as follows: 2 mg/mlL Biotin, 2mg/L Folic acid, 10mg/L vitamin B65mg/L vitamin B15mg/L vitamin B25mg/L nicotinic acid, 5mg/L pantothenic acid, 0.1mg/L vitamin B125mg/L aminobenzoic acid and 5mg/L lipoic acid.
CO reduction by adopting the device2The process for methane is as follows:
1.5mL (1.5%) of methanogen pasteurella (Methanosarsina bakeri) solution was added to the cathode pool, and the headspace was filled with CO2And N2Mixed gas (1: 4). The working electrode (i.e. the cathode 4) is made of a nitric acid-acetic acid mixed fiber film attached to a multi-wall carbon nano tube. Firstly, a multi-wall carbon nano tube with the length of 50 mu m is soaked in 5 percent hydrogen peroxide and shaken for 1h, and then the multi-wall carbon nano tube is dissolved in 15 percent nafion ethanol solution after being dried. And performing ultrasonic treatment for 20min to uniformly disperse the multi-walled carbon nanotubes to prepare a multi-walled carbon nanotube nafion ethanol solution. Then uniformly coating the mixture on a nitric acid-acetic acid mixed fiber film with the aperture of 1.5 mu m, and drying the film in an environment of 55 ℃. The loading capacity of the multi-wall carbon nano-tube of the nitric acid-acetic acid mixed fiber film is 8mg/cm2. The working electrode was placed horizontally, and the potential of the working electrode was adjusted to-0.6V (vs SHE) to carry out the reaction.
Control group B was inoculated with 1.5mL (1.5%) of methanogen pasteurella (Methanosarsina bakeri) solution in the cathode pool, and the headspace was filled with CO2And N2Mixed gas (1: 4). The working electrode adopts a traditional carbon cloth electrode, and the potential of the working electrode is adjusted to-0.6V (vs SHE) for reaction.
Control group C was inoculated with 1.5mL (1.5%) of methanogen pasteurella (methanosarina bikeri) in a cathode cell without placing a working electrode and applying an external potential. Headspace filled with CO2And N2Mixed gas (1: 4).
The headspace gas composition of the three experiments was determined using gas chromatography after 78 hours. As a result, 8.453. mu. mol was found in group A, and 6.371. mu. mol was found in group B. Group C methane production was 3.379. mu. mol. It can be seen that the present invention significantly improves methane production under the same conditions. Has wide application prospect.
Example 2:
this example was carried out simultaneously in experimental group a and control groups B, C:
experimental group a is shown in figure 1.5mL (5.0%) of methanogen brucellosis bacterial solution (Methanobacterium byrantti) is inoculated into the cathode pool, and the headspace is filled with CO2And N2Mixed gas (1: 4). The working electrode is made of a nitric acid-acetic acid mixed fiber film attached to a multi-wall carbon nano tube. Firstly, a multi-walled carbon nanotube with the length of 10 mu m is soaked in 5 percent hydrogen peroxide and shaken for 1h, and then is dissolved in 2 percent nafion ethanol solution after being dried. And performing ultrasonic treatment for 1h to uniformly disperse the multi-walled carbon nano-tubes to prepare a multi-walled carbon nano-tube nafion ethanol solution. Then evenly coating the mixture on a nitric acid-acetic acid mixed fiber film with the aperture of 2 mu m, and drying the film in an environment of 37 ℃. The loading capacity of the multi-wall carbon nano-tube of the nitric acid-acetic acid mixed fiber film is 4.5mg/cm2. The potential of the working electrode was adjusted to-0.6V (vs SHE) to effect the reaction.
Control group B was inoculated with 5mL (5.0%) of methanogen brucellosis (Methanobacterium byrantti) solution in the cathode cell, and the headspace was filled with CO2And N2Mixed gas (1: 4). The working electrode adopts a traditional carbon cloth electrode, and the potential of the working electrode is adjusted to-0.6V (vs SHE) for reaction.
In the control group C, 5mL (5.0%) of methanogen brucellosis bacterial solution (Methanobacterium byrantti) was inoculated into the cathode pool, and no working electrode was placed and no external potential was applied. Headspace filled with CO2And N2Mixed gas (1: 4).
The headspace gas composition of the three experiments was determined using gas chromatography after 78 hours. As a result, 7.471. mu. mol was found in group A, and 5.643. mu. mol was found in group B. Group C methane production was 1.808. mu. mol. It can be seen that the present invention significantly improves methane production under the same conditions. Has wide application prospect.
Example 3:
this example was carried out simultaneously in experimental group a and control groups B, C:
experimental group a is shown. 3mL (3.0%) of methanogen hennerii liquid (Methanospirilum hungatei) was inoculated into the cathode pool, and the headspace was filled with CO2And N2Mixed gas (1: 4). The working electrode adopts the attachmentA nitric acid-acetic acid mixed fiber film coated with multi-wall carbon nano-tubes. Firstly, a multi-walled carbon nanotube with the length of 2 mu m is soaked in 5 percent hydrogen peroxide and shaken for 1h, and then the multi-walled carbon nanotube is dissolved in 20 percent nafion ethanol solution after being dried. And performing ultrasonic treatment for 5min to uniformly disperse the multi-walled carbon nanotubes to prepare a multi-walled carbon nanotube nafion ethanol solution. Then uniformly coating the mixture on a nitric acid-acetic acid mixed fiber film with the aperture of 0.44 mu m, and drying the film in an environment of 37 ℃. The loading capacity of the multi-wall carbon nano-tube of the nitric acid-acetic acid mixed fiber film is 6.8mg/cm2. The potential of the working electrode was adjusted to-0.6V (vs SHE) to effect the reaction.
Control group B was inoculated with 3mL (3.0%) of methanogen henneri (Methanospirilum hungatei) solution in the cathode pool, and the headspace was filled with CO2And N2Mixed gas (1: 4). The working electrode adopts a traditional carbon cloth electrode, and the potential of the working electrode is adjusted to-0.6V (vs SHE) for reaction.
In control group C, 3mL (3.0%) of methanogen hennerii (Methanospirilum hungatei) was inoculated into the cathode cell, and no working electrode was placed and no external potential was applied. Headspace filled with CO2And N2Mixed gas (1: 4).
The headspace gas composition of the three experiments was determined using gas chromatography after 78 hours. As a result, 6.845. mu. mol was found in group A, and 5.331. mu. mol was found in group B. Group C produced 1.664. mu. mol of methane. It can be seen that the present invention significantly improves methane production under the same conditions. Has wide application prospect.
Claims (6)
1. A method for producing methane by reducing carbon dioxide with microorganisms is characterized by comprising the following steps:
1) constructing a microbial electrolysis cell comprising a cathode cell and an anode cell, wherein the cathode cell comprises a working electrode and a reference electrode; the anode cell includes a counter electrode; the cathode pool and the anode pool are separated by a proton exchange membrane; the working electrode is made of a nitric acid-acetic acid mixed fiber film adhered to the multi-wall carbon nano tube; the amount of the multi-walled carbon nano-tubes adhered to the nitric acid-acetic acid mixed fiber film is 0.1-10 mg/cm2The length of the multi-wall carbon nanotube is 0.5-50 μm; nitric acidThe aperture of the acetic acid mixed fiber film is 0.2-2 mu m;
2) adding methanogen culture medium into the cathode pool and the anode pool;
3) inoculating 1.5-5.0% methanogen bacteria liquid in a cathode pool, and culturing;
4) loading-0.6 v vs SHE voltage to the cathode pool;
5) methane was harvested from the headspace of the cathode cell.
2. The method for producing methane by reducing carbon dioxide with microorganisms according to claim 1, wherein the proton exchange membrane in step (1) is an N117 proton exchange membrane.
3. The method for producing methane by reducing carbon dioxide with microorganisms according to claim 1, wherein the formulation of the methanogen culture medium in step (2) is as follows: 0.35g/L K2HPO4，0.23g/L KH2PO4，0.5g/L NH4Cl，0.5g/L MgSO4·7H2O，0.25 g/L CaCl2，2.25g/L NaCl，0.85g/L NaHCO3，0.5g/L Na2S·H2O, 10ml/L of trace element solution and 1ml/L of vitamin solution; the head space of the electrolytic cell is CO2:N2A mixed gas of =1:4 (n/n).
4. The method of claim 3, wherein the formulation of the trace element solution is as follows: 1500mg/L FeCl2·4H2O，70mg/L ZnCl2，100mg/L MnCl2·4H2O，6mg/L H3BO3，190 mg/L CoCl2·6H2O，2mg/L CuCl2·H2O，24mg/L NiCl2·6H2O， 36 mg/L NaMo4·2H2O，25%HCl10ml/L；
The formula of the vitamin solution is as follows: 2mg/L biotin, 2mg/L folic acid, 10mg/L vitamin B65mg/L vitamin B15mg/L vitamin B25mg/L nicotinic acid, 5mg/L pantothenic acid0.1mg/L vitamin B125mg/L aminobenzoic acid and 5mg/L lipoic acid.
5. The method for producing methane by reducing carbon dioxide with microorganisms according to claim 1, wherein the methanogen in step (3) comprises methanogen pasteurianus (methane:)Methanosarcina barkeri) Methanogen Brucella (a)Methanobacterium byrantti) And methanogen henle: (Methanospirllum hungatei）。
6. The method for producing methane by reducing carbon dioxide with microorganisms according to claim 1, wherein the methanogen in step (3) is cultured at room temperature.
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| DE102020123184A1 (en) | 2020-09-04 | 2022-03-10 | Electrochaea GmbH | Process for the production of a synthetic product, e.g., methane, using methanogenic microorganisms in a microbial electrolytic cell (MEC) by employing a separate nutrient supply strategy |
| CN113564628B (en) * | 2021-08-13 | 2022-12-30 | 中国科学技术大学 | Metal-free molecular catalyst and preparation method and application thereof |
| CN114699908A (en) * | 2022-01-20 | 2022-07-05 | 中国环境科学研究院 | Activated sludge coupled device and method for fixing carbon dioxide by driving microorganisms with electric energy |
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