CN114107423A - Method for promoting microbial hydrogen-alkane conversion based on porous material - Google Patents
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Abstract
The invention relates to the field of biological energy, in particular to a method for promoting microbial hydrogen-alkane conversion based on a porous material. The porous material treated by 1-10% of dilute hydrochloric acid is used as a biological carrier, the HYTHANE conversion composite microbial inoculum is grown and propagated on the surface of the pretreated porous material to form a stable biological membrane, and then the biological carrier forming the biological membrane is used for quickly converting carbon dioxide in the methane into methane. The porous material-composite microbial inoculum biomembrane complex can realize the rapid conversion of carbon dioxide and exogenous hydrogen in the biogas into methane, so that the concentration of the methane in the biogas is increased to more than 98 percent, and the production rate of the methane is increased by more than 70 percent compared with the conventional suspension culture method. The invention not only greatly improves the heat value of the biogas and enhances the practicability of the biogas, but also can reduce the emission of a large amount of carbon dioxide. Thus, the practice of the present process results in good economic and environmental benefits.
Description
Technical Field
The invention relates to the field of biological energy, in particular to a method for promoting microbial hydrogen-alkane conversion based on a porous material.
Background
In recent years, the proportion of green renewable energy in energy utilization is increasing. The key technology of the method is to prepare the biogas by utilizing the anaerobic fermentation of organic wastes or energy crops. However, the methane content of biogas is usually only 50% to 70% and the carbon dioxide content 30% to 50%. Therefore, how to effectively increase the methane content in the biogas is an urgent problem to be solved. At present, the removal of carbon dioxide from biogas mainly depends on some physical methods, such as pressure swing adsorption, membrane separation, etc., which are effective but too costly. At the same time, carbon dioxide is discharged directly into the atmosphere, which not only causes resource waste, but also aggravates the greenhouse effect.
The short-time 'surplus' wind power or solar power is used for electrolyzing water into hydrogen and oxygen, and the generated hydrogen and carbon dioxide produced by biogas engineering are used for generating methane through microbial reaction, so that the method is a hotspot field of biogas purification at present. Patents CN 103113010A, CN108265081A, CN 205152232U, CN 204589159U and so on propose the whole process flow from hydrogen preparation to microbial conversion of carbon dioxide into methane, however, these patents cannot solve the bottleneck problem of low hydrogen-methane conversion efficiency in the current biogas purification. Although patent publication No. CN103958688A proposes a technique for improving the conversion of carbon dioxide into methane by using a hollow fiber membrane bioreactor, the hollow fiber membrane has problems of high cost and easy blockage during operation, which limits its commercial application. It is noted that in the above patent, the hydrogen-trophic methanogens used in the core process unit portion for the conversion of hydrogen and carbon dioxide to methane are all cultured in suspension. The technology has the defects that functional active microorganisms are easy to lose, so that the hydrogen-methane conversion efficiency is low and the methane concentration is low.
Therefore, in combination with the above problems, the present invention has developed a method for promoting microbial hydrogen alkane conversion based on porous materials. The method realizes the rapid reaction of the exogenously introduced hydrogen and carbon dioxide to generate methane, and solves the technical problem of low hydrogen-alkane conversion efficiency caused by easy loss of functional active microorganisms and the like.
Disclosure of Invention
The invention aims to provide a method for promoting microbial hydrogen-alkane conversion based on a porous material.
In order to achieve the purpose, the invention adopts the technical scheme that:
a method for promoting microbial hydrogen-alkane conversion based on a porous material adopts a porous material (such as biochar, ceramsite and the like) treated by 1-10% of dilute hydrochloric acid as a biological carrier, a hydrogen-alkane conversion composite microbial inoculum grows and breeds on the surface of the pretreated porous material to form a stable biological film, and then the biological carrier forming the biological film is utilized to rapidly convert carbon dioxide in methane into methane. The composite can quickly convert carbon dioxide in the biogas into methane, and the concentration of the methane in the biogas can be increased to more than 98%. The methane yield is improved by more than 70 percent compared with the conventional suspension culture mode.
The compound hydrogen alkane converting microbial inoculum consists of two floras in a cooperative relationship, namely volatile fatty acid oxidizing bacteria responsible for degrading small molecular volatile fatty acids such as acetic acid, propionic acid and butyric acid in the hydrogen alkane converting process and hydrogenotrophic methane bacteria responsible for converting hydrogen and carbon dioxide into methane; wherein the fatty acid oxidizing bacteria comprise Petrimoas and Geobacter (each 50% by mass); the hydrogenotrophic Methanobacterium is composed of Methanobacterium and methanorebabacter (each 50% by mass).
Mixing the volatile fatty acid oxidizing bacteria and the hydrogenotrophic methane bacteria according to the mass ratio of 1: 1; wherein, the Petrimoas and the Geobacter are mixed according to the mass ratio of 1: 1; methanobacterium and Methanobrevibacter are mixed according to the mass ratio of 1: 1.
The biological carrier is prepared by treating a porous material with 1-10% diluted acid for 12-24h, washing with deionized water to about pH7, and keeping at 121 deg.C for 30 min.
Further, the pretreated porous material, the culture medium and the compound microbial inoculum concentrated solution obtained after centrifugation are mixed; wherein the porous material and the concentrated solution respectively account for 10-30% of the culture medium by mass. Then controlling the introduction ratio of hydrogen to carbon dioxide to be 4:1(v/v), the total gas flow is 1-5ml per liter working volume per minute, and the temperature required for culture is 35 ℃. Culturing for 36-72h, and forming a microbial film on the surface of the porous material by the composite microbial inoculum.
The culture medium comprises, by mass, 1-2% of calcium carbonate, 1-3% of potassium dihydrogen phosphate, 1-2% of disodium hydrogen phosphate, 1-2% of ammonium chloride, 1-3% of yeast powder and the balance of water, and the pH value is 7-8. Sterilizing at 121 deg.C for 30 min.
The compound microbial inoculum concentrated solution obtained after centrifugation is prepared by inoculating a freeze-dried compound microbial inoculum (namely strains mixed according to the proportion) in the culture medium after activation (at the culture temperature of 35 ℃), inoculating a primary culture in a fermentation tank (1L), and culturing for 36-72 hours at 37 ℃ under the condition that the proportion of hydrogen and carbon dioxide is 4:1 (v/v). When OD600 reaches about 0.5, the compound bacteria culture solution is centrifuged and concentrated for later use.
The composite microbial inoculum biomembrane-porous material complex is transferred into a hydrogen alkane conversion reactor (1-2L) and filled to 2/3 parts of the volume of the reactor, mixed gas of methane (the concentration of carbon dioxide is 45-50%) and hydrogen is introduced from the bottom of the reactor, the introduction ratio of the hydrogen and the carbon dioxide is controlled to be 4:1(v/v), the flow rate is controlled to be 1-5ml/min, and continuous and stable methane production is carried out.
The invention has the following advantages:
1. the invention directly adopts hydrogen and methane as raw material gas to produce high-concentration biological methane, realizes energy regeneration and high-value utilization of carbon dioxide in methane components, and improves the heat value of the methane.
2. The composite microbial inoculum biomembrane-porous material complex adopted by the invention enables the composite microbial flora to be stably fixed on the biological carrier in the form of a biomembrane, thereby reducing the loss of microbial biomass and improving the hydrogen-alkane conversion efficiency.
3. The invention can utilize a composite microbial agent biological membrane-porous material composite system, and the affinity of carbon dioxide and hydrogen molecules is improved due to the introduction of active functional groups into the pretreated porous material. The composite microbial inoculum biomembrane attached to the surface of the porous material can quickly convert hydrogen and carbon dioxide into methane.
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Fig. 1 is a scanning electron microscope image of a composite microbial agent biofilm-biochar complex provided by an embodiment of the invention, wherein a is untreated biochar; and B, treating the biochar with acid.
Detailed Description
The following examples are presented to further illustrate embodiments of the present invention, and it should be understood that the embodiments described herein are for purposes of illustration and explanation only and are not intended to limit the invention.
The invention adopts porous materials (such as biochar, ceramsite and the like) as biological carriers, facilitates the growth and the propagation of hydrogenotrophic methanogens, can quickly convert carbon dioxide into methane by injecting a proper amount of hydrogen, and can improve the concentration of methane in the methane to more than 90 percent through the conversion of the hydrogenotrophic methanogens.
Porous materials (such as biochar, ceramsite and the like) are used as biological carriers, and active functional groups are formed after acid treatment, so that the affinity of hydrogen and carbon dioxide molecules is improved. The hydrogen-alkane conversion complex microbial inoculum grows and breeds on the surface of the hydrogen-alkane conversion complex microbial inoculum, and can form a stable biological membrane. The composite can quickly convert carbon dioxide in the biogas into methane, the conversion efficiency of the hydrogen and the methane in the biogas is more than 98%, and the concentration of the methane can be increased to more than 98%. The methane yield is improved by more than 70 percent compared with the conventional suspension culture mode.
In a still further aspect of the present invention,
(1) the microbial agent comprises the following components: the compound microbial inoculum consists of volatile fatty acid oxidizing bacteria, hydrogenotrophic methanobacteria and 2 large floras (the mass of each bacterium is 50%). Wherein the fatty acid oxidizing bacteria are responsible for degrading small-molecule volatile fatty acids such as acetic acid, propionic acid and butyric acid in the process of hydrogen alkane conversion and consist of Petrimoas and Geobacter (the mass is 50 percent respectively); the hydrogenotrophic methanogens, which are responsible for converting hydrogen and carbon dioxide into methane, are composed of Methanobacterium and methanorevibacter (50% by mass each). 2 floras are in a cooperative relationship, and are absent, and the fatty acid generated in the hydrogen alkane conversion process is cooperatively and further converted into methane.
(2) And (5) culturing. The formula of the complex microbial inoculum culture medium comprises: 1-2% of calcium carbonate, 1-3% of monopotassium phosphate, 1-2% of disodium hydrogen phosphate, 1-2% of ammonium chloride, 1-3% of yeast powder and water. The pH value is 7-8. Sterilizing at 121 deg.C for 30 min.
And (3) centrifuging to obtain a compound microbial inoculum concentrated solution: the freeze-dried compound microbial inoculum is taken, activated in the culture medium (culture temperature is 35 ℃), the primary culture is taken, inoculated into a fermentation tank (1L), and cultured for 36-72 hours at 37 ℃ under the condition that the ratio of hydrogen to carbon dioxide is 4: 1. When OD600 reaches about 0.5, the compound bacteria culture solution is centrifuged and concentrated for later use.
(3) And (4) film forming. After porous materials such as biochar, ceramsite and the like are treated by 1-10% diluted acid (for 12-24 hours), impurities on the surface are removed, and meanwhile, active functional groups are formed on the surface of the porous materials, so that the adsorption efficiency of hydrogen and carbon dioxide is improved. And washing the acid-treated porous material with deionized water to about pH7 at 121 ℃ for 30min for later use. Putting the treated porous material into a reactor (1-5L), adding the culture medium, and adding the complex microbial inoculum concentrate, wherein the ratio of the complex microbial inoculum concentrate to the porous material is controlled at 10-30%. Then controlling the introduction ratio of hydrogen to carbon dioxide to be 4:1, the total gas flow is 1-5ml per liter working volume per minute, and the temperature required for culture is 35 ℃. Culturing for 36-72h to complete the film forming process of the composite microbial inoculum on the surface of the porous material.
(4) And (4) hydrogen alkane conversion. Transferring the composite microbial inoculum biomembrane-porous material complex into a hydrogen alkane conversion reactor (1-2L), and filling the reactor 2/3. Introducing a mixed gas of biogas (the concentration of carbon dioxide is 45-50%) and hydrogen from the bottom of the reactor, controlling the introduction ratio of hydrogen and carbon dioxide at 4:1(v/v) and the flow rate at 1-5 ml/min. Continuous and stable methane production is carried out. The hydrogen-methane conversion efficiency of the reactor is more than 98 percent, and the methane concentration is more than 99 percent.
Example (biochar):
(1) and (4) pretreating the biochar. Treating the biochar porous material for 12 hours by using dilute hydrochloric acid with the concentration of 10 wt%, removing impurities on the surface, and forming active functional groups on the biochar surface; and then washing the acid-treated porous material by deionized water to about pH7, and keeping the porous material at 121 ℃ for 30min for later use.
(2) And (4) film forming. Respectively putting biochar treated for 12 hours or biochar which is not pretreated into different reactors (1L), adding 500mL of complex microbial inoculum culture medium into each reactor, and then adding activated complex microbial inoculum concentrated solution; the pretreated biochar porous material and the activated complex microbial inoculum concentrated solution respectively account for 15% (w/w) of the mass of the culture medium. Controlling the introduction ratio of hydrogen to carbon dioxide to be 4:1(v/v), total gas flow of 2ml per liter working volume per minute, and temperature required for culture of 35 ℃. Culturing for 48h, and respectively finishing the complex of the composite microbial inoculum in the film forming process on the surfaces of different porous materials (see figure 1).
The composite microbial inoculum consists of 2 large floras (the content is 50wt percent respectively) such as volatile fatty acid oxidizing bacteria, hydrogenotrophic methane bacteria and the like. Wherein the fatty acid oxidizing bacteria are responsible for degrading small-molecule volatile fatty acids such as acetic acid, propionic acid, butyric acid and the like in the process of hydrogen alkane conversion and consist of Petrimoas and Geobacter (the content is 50wt percent respectively); the hydrogenotrophic methanogens, which are responsible for converting hydrogen and carbon dioxide into methane, are composed of Methanobacterium and methanorevibacter (each 50 wt%). 2 floras are in a cooperative relationship, and are absent, and the fatty acid generated in the hydrogen alkane conversion process is cooperatively and further converted into methane.
A composite microbial inoculum culture medium: according to weight percentage, 1 percent of calcium carbonate, 3 percent of potassium dihydrogen phosphate, 2 percent of disodium hydrogen phosphate, 2 percent of ammonium chloride, 1 percent of yeast powder and the balance of water; pH 7; sterilizing at 121 deg.C for 30 min.
Activated complex microbial inoculum concentrated solution: the freeze-dried compound bacterial agent (i.e. the bacterial strains mixed according to the proportion) is taken, activated in the culture medium (the culture temperature is 35 ℃), the primary culture is taken, inoculated into a fermentation tank (1L), and cultured for 36-72 hours at 37 ℃ under the condition that the proportion of hydrogen and carbon dioxide is 4: 1. When OD600 reaches about 0.5, the compound bacteria culture solution is centrifuged and concentrated for later use.
(4) And (4) hydrogen alkane conversion. Transferring the two composite microbial inoculum biomembrane-biochar complexes into a hydrogen alkane conversion reactor (1L), and filling the reactor with the volume 2/3 by taking an unadditized carrier as a control. Introducing a mixed gas of biogas (the concentration of carbon dioxide is 45%) and hydrogen from the bottom of the reactor, controlling the introduction ratio of hydrogen and carbon dioxide at 4:1(v/v) and the flow rate at 2 ml/min. Continuous and stable methane production is carried out.
As shown in FIG. 1B, the complex bacteria are uniformly distributed on the surface of the biochar, and a stable structure of the biofilm is formed. After the acid treatment, the biological film is more compact and firm, and microorganisms are difficult to fall off.
TABLE 1 comparison of methane yield (L/L/d) after addition of biochar support
| Continuous hydroalkanes conversion stage | Control group | Untreated biochar supports | Acid treated biochar supports |
| 1 | 0.032±0.006 | 0.090±0.005 | 0.143±0.023 |
| 2 | 0.648±0.140 | 0.945±0.032 | 1.151±0.015 |
| 3 | 0.700±0.080 | 0.938±0.011 | 1.262±0.007 |
| 4 | 0.647±0.033 | 1.164±0.042 | 1.365±0.026 |
| 5 | 0.607±0.118 | 1.033±0.035 | 1.269±0.004 |
As shown in the data of 5 stages in the operation process in the table 1, the average methane yield of the untreated biochar group can reach 1.164L/L/d to the maximum, the methane yield is improved by 80 percent compared with that of a control group in the same period, and the methane concentration reaches more than 98 percent; the average methane yield of the acid-treated biochar is improved by 31 percent compared with that of the untreated biochar. Proves that the biochar after acid treatment has better effect.
Claims (6)
1. A method for promoting microbial hydrogen-alkane conversion based on a porous material is characterized by comprising the following steps: the porous material treated by 1-10% of dilute hydrochloric acid is used as a biological carrier, the HYTHANE conversion composite microbial inoculum is grown and propagated on the surface of the pretreated porous material to form a stable biological membrane, and then the biological carrier forming the biological membrane is used for quickly converting carbon dioxide in the methane into methane.
2. The method of claim 1 for promoting microbial HYTHANE CONVERSION BASED ON A POROUS MATERIAL, wherein: the compound hydrogen alkane converting microbial inoculum consists of two floras in a cooperative relationship, namely volatile fatty acid oxidizing bacteria responsible for degrading small molecular volatile fatty acids such as acetic acid, propionic acid and butyric acid in the hydrogen alkane converting process and hydrogenotrophic methane bacteria responsible for converting hydrogen and carbon dioxide into methane; wherein the fatty acid oxidizing bacteria comprise Petrimoas and Geobacter; the methanogens hydrogenotrophic bacteria consist of Methanobacterium and methanobacterbrevibacter.
3. The method of claim 1 for promoting microbial HYTHANE CONVERSION BASED ON A POROUS MATERIAL, wherein: the biological carrier is prepared by treating a porous material with 1-10% diluted acid for 12-24h, washing with deionized water to about pH7, and keeping at 121 deg.C for 30 min.
4. A method for promoting microbial HYTHANE CONVERSION BASED ON A POROUS MATERIAL as claimed in any one of claims 1 to 3, wherein: and (3) carrying out pretreatment on the porous material, the culture medium and the compound microbial inoculum concentrated solution obtained after centrifugation, and then controlling the introduction ratio of hydrogen to carbon dioxide to be 4:1(v/v), the total gas flow is 1-5ml per liter working volume per minute, and the temperature required for culture is 35 ℃. Culturing for 36-72h, and forming a microbial film on the surface of the porous material by the composite microbial agent;
wherein, the pretreated porous material and the compound microbial inoculum concentrated solution obtained after centrifugation respectively account for 10-30% of the mass of the culture medium.
5. The method of claim 4 for promoting microbial HYTHANE CONVERSION BASED ON A POROUS MATERIAL, wherein: the culture medium comprises, by mass, 1-2% of calcium carbonate, 1-3% of potassium dihydrogen phosphate, 1-2% of disodium hydrogen phosphate, 1-2% of ammonium chloride, 1-3% of yeast powder and the balance of water, and the pH value is 7-8.
6. The method of claim 4 for promoting microbial HYTHANE CONVERSION BASED ON A POROUS MATERIAL, wherein: the composite microbial inoculum biomembrane-porous material complex is transferred into a hydrogen alkane conversion reactor (1-2L) and filled to 2/3 parts of the volume of the reactor, mixed gas of methane (the concentration of carbon dioxide is 45-50%) and hydrogen is introduced from the bottom of the reactor, the introduction ratio of the hydrogen and the carbon dioxide is controlled to be 4:1(v/v), the flow rate is controlled to be 1-5ml/min, and continuous and stable methane production is carried out.
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| JP2010239962A (en) * | 2009-03-19 | 2010-10-28 | Central Res Inst Of Electric Power Ind | Method for producing methane gas using carbon dioxide |
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| JP2010239962A (en) * | 2009-03-19 | 2010-10-28 | Central Res Inst Of Electric Power Ind | Method for producing methane gas using carbon dioxide |
| US20150099286A1 (en) * | 2012-05-08 | 2015-04-09 | Maa-Ja Elintarviketalouden Tutkimuskeskus | Means and methods for methane production |
| KR20190091947A (en) * | 2018-01-30 | 2019-08-07 | 건국대학교 산학협력단 | Macro-porous particles for immobilization of Methanotrophs and methode for production of methanol using the same |
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