CN110042042A - A kind of the methane-oxidizing bacteria domestication culture apparatus and method of simulation underground coal mine environment - Google Patents

A kind of the methane-oxidizing bacteria domestication culture apparatus and method of simulation underground coal mine environment Download PDF

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CN110042042A
CN110042042A CN201910327803.0A CN201910327803A CN110042042A CN 110042042 A CN110042042 A CN 110042042A CN 201910327803 A CN201910327803 A CN 201910327803A CN 110042042 A CN110042042 A CN 110042042A
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谢军
辛林
程卫民
杜海刚
宋玥
付海政
王怡
任广意
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Shandong University of Science and Technology
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Abstract

The invention discloses a kind of methane-oxidizing bacteria domestication culture apparatus for simulating underground coal mine environment and methods.Including raw coal stress loading part, environment temperature control section, environmental gas atmosphere injection part, methane gas injection part, methane oxidation bacterium solution injection unit point, methane gas on-line monitoring part, environment gas chromatographic analysis part, environment liquid phase mass spectrometric analysis part and nine part of the control section environment pH composition.The beneficial effects of the invention are as follows the microorganism enrichment states for capableing of accurate and effective simulation underground coal mine condition, simulate stress of coal seam, temperature, pH, atmosphere condition.

Description

A kind of the methane-oxidizing bacteria domestication culture apparatus and method of simulation underground coal mine environment
Technical field
The invention belongs to technical field of coal exploitation, are related to a kind of methane-oxidizing bacteria domestication training for simulating underground coal mine environment Support device and method.
Background technique
China is a developing country, and the energy is very in short supply, and coal accounts for 70% or more of China's energy total flow, coal The yield of charcoal directly affects the development of national economy, as long as which results in the place for having coal reserve in China, it is either low Gas or highly gas mining area are all opened a mine energetically creating conditions.And all coal mines in China are gaseous mine, according to statistics, In large and medium-sized coal mine, highly gassy mine accounts for 20.34%, and projecting mine accounts for 19.77%.In small-size coal mine, highly gassy mine is accounted for 15% or so.This is one of the reason for causing Gas Disaster more.Only during 2007~2016 years, all kinds of gas occur altogether for the whole nation Disaster accident 389 rises, dead 3307 people.Wherein, gas explosion accident 196 rises, accounting 50%；2022 people of death toll, accounting 61%.Coal and gas prominent accident 150 rises, accounting 39%；1084 people of death toll, accounting 33%.Gas poisoning and asphyxia thing Therefore 43, accounting 11%；201 people of death toll, accounting 6%.Gas Disaster causes huge economic loss and casualties, Negative effect is brought to economic development and social stability.Gas Disaster seriously threatens the safety in production of coal mine, wherein coal with Gas Outburst is particularly acute, while being also increasingly taken seriously to the improvement of Gas Outburst.Currently, China's coal-mine gas is main Treatment process is that the synthesis of the prediction of coal seam gas-bearing capacity and outburst amount, mine ventilation, mine gas drainage and quaternity is anti- Prominent measure.But China Mine Geological condition and the coal mine own characteristic formed therefrom determine existing gas drainage under suction skill The limitation of art.Since primitive coalbed gas bearing capacity is big, the original poor air permeability in coal measure strata coal seam causes primitive coalbed to take out in advance It is difficult to carry out, and effect is very poor.If can be generated to environment very big in addition, the gas that extraction is gone out can be discharged into environment Adverse effect.Secondly, the main blower of mine and the Design of ventilation ability of ventilation network are limited, at quite long one section It is constant in phase, thus also have limitation by ventilation exclusion gas.In recent years, with the development of science and technology utilizing chemistry and micro- life The technology that object administers gas achieves greater advance.Wherein, adiabatic oxygenation method passes through thermal decomposition, Catalytic Thermal Decomposition and heat of adsorption point Solution combines, can be by the CH in gas4CO is oxidized to CO2And H2The pollution-free mixture of O, to make near-by explosion concentration Methane gas and oneself up to explosion ratio methane gas be diluted to explosion ratio hereinafter, but because this method to device requirement compared with Height, and pre-dilution is required to high concentration gas, operating process is complicated, and higher cost is unfavorable for large-scale industrial application.It compares For, the method practicability of microbial treatment gas is stronger, is adopted by the methane-oxidizing bacteria degradation being widely present using nature Dead zone and coal-bed gas reduce the gas emission in seam mining, inhibit coal mine gas accumulation, or make high concentration gas Gas drop to gas explosion lower limit in a short time hereinafter, can to other administer gas measure play booster action, from And achieve the purpose that improve safety of coal mines.Methane-oxidizing bacteria is very extensive in distributed in nature, fills out from sludge, marsh, rubbish Burying can be isolated in the environment such as field, river, fallen leaves, sewer, paddy field, ocean, pond, range land soil.However, coal Coal in layer is the substance for having multiple-void structure and very strong adsorptivity, is unminded in underground lower in high-pressure state, device in Gas Body is existed in the form of ADSORPTION STATE, rather than is existed with free state, when exploitation, exposes work surface pressure reduction, gas with Gas form is released.In addition to the difference of existence and condition of high voltage, coal seam is a kind of state of anoxic again, and in coal seam Also preservation CO, CO2And H2S etc. breeds harmful gas to methane oxidizing bacteria growth.Therefore, a kind of simulation coal mine is invented in research The methane-oxidizing bacteria domestication culture apparatus and method of subsurface environment, the items such as experiment different temperatures, atmosphere, stress of coal seam and pH Coal seam conditions under part screen, tame out and can adapt to the mining as the amphimicrobian type of primary carbon source using methane of coal mine environment Methane-oxidizing bacteria has vital meaning.
The screening and domestication of mining methane-oxidizing bacteria are an extremely complex Bioexperiment processes, and essence is exactly one logical The living environment for simulating down-hole coal bed occurrence status to build methane-oxidizing bacteria is crossed, to realize to the mine for being suitable for coal production Use methane-oxidizing bacteria.The key of the process has: first is that simulation ocurrence of coal seam state；Second is that simulation coal seam temperature；Third is that simulation coal Layer atmosphere.Hole fissured structure and its very strong adsorptivity due to coal, appearance of the device in Gas far more than coal seam itself hole Product.Therefore under the effect of larger pressure, coal-bed gas exists with another state, that is, ADSORPTION STATE.In addition to existence and high-pressure section The difference of part, coal seam are a kind of state of anoxic again, and also preservation CO, CO in coal seam2And H2S etc. is to methane oxidizing bacteria growth Breed harmful gas.Wherein, the simulation of underground atmosphere is the committed step and shadow of methane-oxidizing bacteria screening and domestication Ring the important factor in order of methane-oxidizing bacteria underground adaptability.Therefore, the stress to methane-oxidizing bacteria in coal seam, temperature, pH, Atmosphere carries out comprehensive analog simulation, makes coal production environment true reappearance, to realize the mining methane oxygen of high-adaptability Change Effective selection and the domestication work of bacterium.And the experimental provision of existing methane-oxidizing bacteria degradation raw coal adsorption gas cannot be effective Simulate the pH and atmosphere in coal seam.
Summary of the invention
The purpose of the present invention is to provide a kind of methane-oxidizing bacteria domestication culture apparatus for simulating underground coal mine environment and sides Method.Technical solution used by apparatus of the present invention be include raw coal stress loading part, environment temperature control section, environmental gas Atmosphere injection part, methane gas injection part, methane oxidation bacterium solution injection unit point, methane gas on-line monitoring part, environment Gas chromatographic analysis part, environment liquid phase mass spectrometric analysis part and nine part of the control section environment pH composition.
Raw coal stress loading part, which specifically includes that, calculates meter, oil pressure pump, pressure controller, pressure sensor and pressure Force detector.
The environment temperature control section includes: water bath heating device and bath temperature monitor.
The environmental gas atmosphere injection unit point includes: hydrogen sulfide gas, CO gas, carbon dioxide gas, valve Door, air inlet valve and air inlet.
The methane gas injection unit point includes: high pressure methane gas, valve, air inlet valve and air inlet.
The methane oxidation bacterium solution injection unit point includes: methane oxidation bacteria liquid, fluid injection valve and liquid injecting pipeline.
Methane gas on-line monitoring part includes: computer, CH4Sensor and CH4Gas monitor.
Environment gas chromatographic analysis part includes: computer, gas chromatograph and gas sampling mouth.
Environment liquid phase mass spectrometric analysis part includes: computer, liquid chromatograph and liquid sample mouth.
The control section the environment pH includes: pH regulating device and valve
A kind of methane-oxidizing bacteria domestication and culture method for simulating underground coal mine environment, comprising the following steps:
Step 1, test coal sample preparation
Coal sample needed for this project is derived from high gas layer, is broken into bulk in laboratory, is put into underground coal mine environmental test Test device physical compression container.
Step 2, coal sample physical compression
Apply certain pressure to coal sample by raw coal stress loading part, simulate reset pressure state locating for coal seam, The reset pressure that this project applies is controlled by oil pressure pump and pressure controller, and it is 20MPa that maximum, which applies pressure,.
Step 3, environment harmful gas injects
Environment harmful gas mainly includes CO, CO2、H2S gas simulates down-hole coal bed atmosphere environment, according to underground coal The real gas atmosphere environment of layer injects a certain amount of environmental gas to device inside coal body.
Step 4, coal sample gas pressure
High gas layer gas pressure is simulated by the methane gas that certain pressure is injected to coal sample in methane gas injection part Power, it is 10MPa that present apparatus methane gas, which injects part highest to apply gas pressure,.
Step 5, environment temperature controls
By environment temperature control system, water bath with thermostatic control control is carried out to temperature locating for the experimental provision external world, makes to test Coal body locating for device coal body is in temperature constant state, and the highest environment temperature which is applied is 50 DEG C.
Step 6, methane oxidation bacterium solution is injected
The methane oxidation bacterium solution of culture is injected into experimental provision coal body by methane oxidation bacterium solution injection pipe, from top Port injection, is dispersed in methane oxidation bacterium solution in coal body.
Step 7, environmental pH is adjusted
By pH regulating device to adjust down-hole coal bed pH value, adjusts it and meet coal seam actual environment pH.
The step 3, step 4, step 5, step 6, step 7 use control variate method, by change atmosphere, pH, The influence of pressure, temperature, methane-oxidizing bacteria night injection quantity research to methane oxidation flora degradation gas ability, analyzes factor change Change the affecting laws for gas ability of breeding and degrade to methane-oxidizing bacteria.
Step 8, methane gas component is monitored on-line
By methane gas monitor on-line part, by Methane monitoring instrument on-line real time monitoring coal body methane gas it is dense Degree leads to the changing rule of methane concentration to analyze methane-oxidizing bacteria resolution gas.
Step 9, environmental gas component monitors
By taken in coal body tracheae connect Outside Vapor chromatograph, analyze environmental interior gas component, concentration changing rule, Especially CO2Concentration variation, to study methane oxidizing bacteria growth process gas release rule.
Step 10, environmental liquids component analysis
By extractor coal body internal solution, coal body internal liquid phase change of component is analyzed using liquid phase mass spectrometric analysis instrument, Analyze the changing rule of methane oxidizing bacteria growth process mesostate；By observing under microscope extracting solution, point Analyse the growth and breeding situation of methane-oxidizing bacteria.
The beneficial effects of the invention are as follows the microorganism enrichment state for capableing of accurate and effective simulation underground coal mine condition, simulations Stress of coal seam, temperature, pH, atmosphere condition.
Detailed description of the invention
Fig. 1 is underground coal mine environmental test test device system figure；
Fig. 2 is analogy method schematic diagram.
In figure: 1- computer；2- liquid chromatograph；3- liquid acquisition mouth；4- gas chromatograph；5- gas collecting mouth；6- Air inlet；7- air inlet valve；8- valve；9- hydrogen sulfide gas；10- CO gas；11- carbon dioxide gas；12- high Press methane gas；13- water bath heating device；14- bath temperature monitor；15- pressure sensor；16- pressure monitor；17- CH4Sensor；18-CH4Gas monitor；19- methane oxidation bacteria liquid；20- fluid injection valve；21- liquid injecting pipeline；22- pressure control Device processed；23- oil pressure pump；24- valve；The coal seam 25-；26-pH control device；27- valve.
Specific embodiment
The present invention is described in detail With reference to embodiment.
Apparatus of the present invention are as shown in Figure 1, include raw coal stress loading part (computer 1, pressure sensor 15, pressure prison Survey device 16, pressure controller 22, oil pressure pump 23), environment temperature control section (water bath heating device 13, bath temperature monitor 14), environmental gas atmosphere injection part (air inlet 6, air inlet valve 7, valve 8, hydrogen sulfide gas 9, CO gas 10, carbon dioxide gas 11), methane gas injection part (air inlet 6, air inlet valve 7, high pressure methane gas 12, valve 24), methane oxidation bacterium solution injection unit point (methane oxidation bacteria liquid 19, fluid injection valve 20, liquid injecting pipeline 21), methane gas are online Monitoring part (computer 1, CH4Sensor 17, CH4Gas monitor 18), environment gas chromatographic analysis part (computer 1, gas Chromatography 4, gas collecting mouth 5), environment liquid phase mass spectrometric analysis part (computer 1, liquid chromatograph 2, liquid acquisition mouth 3) It is formed with the control section environment pH (pH control device 26, valve 27) nine parts.
Raw coal stress loading part specifically includes that calculating meter 1, oil pressure pump 23, pressure controller 22, pressure sensor 15 and pressure detector 16.
The environment temperature control section includes: water bath heating device 13 and bath temperature monitor 14.
The environmental gas atmosphere injection unit point includes: hydrogen sulfide gas 9, CO gas 10, carbon dioxide gas 11, valve 8, air inlet valve 7 and air inlet 6.
The methane gas injection unit point includes: high pressure methane gas 12, valve 24, air inlet valve 7 and air inlet 6.
The methane oxidation bacterium solution injection unit point includes: methane oxidation bacteria liquid 19, fluid injection valve 20 and liquid injecting pipeline 21.
Methane gas on-line monitoring part includes: to calculate meter 1, CH4Sensor 17 and CH4Gas monitor 18.
Environment gas chromatographic analysis part includes: to calculate meter 1, gas chromatograph 4 and gas sampling mouth 5.
Environment liquid phase mass spectrometric analysis part includes: to calculate meter 1, liquid chromatograph 2 and liquid sample mouth 3.
The control section the environment pH includes: pH regulating device 26 and valve 27
Analogy method as shown in Figure 2, comprising the following steps:
Step 1, test coal sample preparation
Coal sample needed for this project is derived from high gas layer, first newly exposes in working face and saws out an approximate square on coal wall Coal column, be sealed with plastic film etc., then lived with the square iron skin mask processed in advance, and guarantee to process axis and run of mine coal seam Reason is vertical, the filled polyurethane in box surrounding gap, finally with wide handsaw that the bottom of square shaped coal cylinder is carefully sawed-off, carries To laboratory, coal sample is carried out to be broken into the random coal cinder of size lumpiness, is put into underground coal mine environmental test test device physics Pressurizing vessel, and coal cinder filled closely knit.
Step 2, coal sample physical compression measure
Apply certain pressure to coal sample by raw coal stress loading part, simulate reset pressure state locating for coal seam, The reset pressure that this project applies is controlled by oil pressure pump 23 and pressure controller 22, keeps certain pressure of methane, maximum is applied Plus-pressure is 20MPa.
Step 3, environment harmful gas injects
Environment harmful gas mainly includes H2S gas 9, CO gas 10, CO2Gas 11 opens air inlet valve 7 and valve 8, according to down-hole coal bed real gas atmosphere environment, gas is injected into coal 25 by air inlet 6, simulates down-hole coal bed gas Atmosphere environment.
Step 4, coal sample gas pressure
High gas layer gas pressure is simulated by the methane gas that certain pressure is injected to coal sample in methane gas injection part Power opens air inlet valve 7 and valve 24, and high pressure methane gas 12 is injected into coal seam 25, present apparatus methane by air inlet 6 It is 10MPa that gas, which injects part highest to apply gas pressure,.
Step 5, environment temperature controls
By environment temperature control system, water bath heating device 13 and water-bath are passed through to temperature locating for the experimental provision external world Temperature monitoring 14 is controlled, and coal body locating for experimental provision coal body is made to be in temperature constant state, the most epipodium which is applied Border temperature is 50 DEG C.
Step 6, methane oxidation bacterium solution is injected
Fluid injection valve 20 is opened, by methane oxidation bacterium solution flow in pipes 21 by a certain amount of methane oxidation bacterium solution of culture 19 are injected into experimental provision coal seam 25, inject from upper port, are dispersed in methane oxidation bacterium solution in coal body.
Step 7, environmental pH is adjusted
By pH regulating device to adjust down-hole coal bed pH value, adjusts it and meet coal seam actual environment pH, adjustable range 5 ~9.
The step 3, step 4, step 5, step 6, step 7 use control variate method, by change atmosphere, pH, The influence of pressure, temperature, methane-oxidizing bacteria night injection quantity research to methane oxidation flora degradation gas ability, analyzes factor change Change the affecting laws for gas ability of breeding and degrade to methane-oxidizing bacteria.
Step 8, methane gas component is monitored on-line
Part is monitored on-line by methane gas, passes through methane gas in 17 on-line real time monitoring coal body of Methane monitoring instrument Concentration leads to the changing rule of methane concentration to analyze methane-oxidizing bacteria resolution gas.
Step 9, environmental gas component monitors
Degradation time is controlled for 24 hours, then by taking tracheae 5 to connect Outside Vapor chromatograph 4 in coal body, is analyzed in environment Portion's gas component, concentration changing rule, especially CO2Concentration variation, to study the release of methane oxidizing bacteria growth process gas Rule.
Step 10, environmental liquids component analysis
By 3 extractor coal body internal solution of liquid acquisition mouth, liquid inside coal body is analyzed using liquid phase mass spectrometric analysis instrument 2 Phase component variation, analyzes the changing rule of methane oxidizing bacteria growth process mesostate；By under microscope to extracting solution It is observed, analyzes the growth and breeding situation of methane-oxidizing bacteria.
Apparatus of the present invention inject H2S、CO、CO2Gas, the true down-hole coal bed atmosphere environment of simulation；Device uses pH Self-checking device, the pH in the down-hole coal bed environment of real simulation.The device is heated using the method for heating water bath, makes first Alkoxide bacterium is heated more evenly, meanwhile, it improves to the safety during coal and heat methane.The device can pass through change The influence of temperature, pH, atmosphere to methane oxidation flora degradation gas ability is investigated in environmental gas atmosphere injection part, thus Obtain environmental factor to the affecting laws of methane-oxidizing bacteria breeding and gas ability of degrading.
It is also an advantage of the present invention that:
(1) the previous deficiency only focused on test and ignore field application is compensated for, it is contemplated that the scene of methane oxidation flora Physical condition；
(2) H is injected2S、CO、CO2Gas, the truer down-hole coal bed atmosphere environment of simulation；
(3) increase pH self-checking device, the more pH in real simulation underground coal mine environment, preferably building culture, Tame the experimental test platform of methane oxidation flora；
(4) water bath heating device is used, methane-oxidizing bacteria is heated evenly, and is improved to coal and heated process gas Safety coefficient.
(5) molecular biological analysis can be carried out to methane oxidizing bacteria growth environment, its feature and function is studied, Explore the rule of methane oxidation flora degradation gas；
The enrichment culture for preferably optimizing methane oxdation mixed bacterial, to obtain the stabilization Mixed Microbes of efficient-decomposition methane Group.
The above is only not to make limit in any form to the present invention to better embodiment of the invention System, any simple modification that embodiment of above is made according to the technical essence of the invention, equivalent variations and modification, Belong in the range of technical solution of the present invention.

Claims (3)

1. a kind of methane-oxidizing bacteria for simulating underground coal mine environment tames culture apparatus, it is characterised in that: add including raw coal stress Carry part, environment temperature control section, environmental gas atmosphere injection part, methane gas injection part, methane oxidation bacterium solution note Enter part, methane gas on-line monitoring part, environment gas chromatographic analysis part, environment liquid phase mass spectrometric analysis part and environment pH Control section.
2. taming culture apparatus according to a kind of methane-oxidizing bacteria for simulating underground coal mine environment described in claim 1, feature exists In: raw coal stress loading part includes: to calculate meter, oil pressure pump, pressure controller, pressure sensor and pressure detector； The environment temperature control section includes: water bath heating device and bath temperature monitor；The environmental gas atmosphere injection unit Dividing includes: hydrogen sulfide gas, CO gas, carbon dioxide gas, valve, air inlet valve and air inlet；The methane Gas injection unit point includes: high pressure methane gas, valve, air inlet valve and air inlet；The methane oxidation bacterium solution injection unit Dividing includes: methane oxidation bacteria liquid, fluid injection valve and liquid injecting pipeline；Methane gas on-line monitoring part include: calculate meter, CH4Sensor and CH4Gas monitor；Environment gas chromatographic analysis part includes: to calculate meter, gas chromatograph and gas Thief hatch；Environment liquid phase mass spectrometric analysis part includes: to calculate meter, liquid chromatograph and liquid sample mouth；The environment pH Control section includes: pH regulating device and valve.
3. existing according to a kind of methane-oxidizing bacteria domestication and culture method for simulating underground coal mine environment, feature described in claim 1 In the following steps are included:
Step 1, test coal sample preparation
Required coal sample is derived from high gas layer, is broken into bulk in laboratory, is put into underground coal mine environmental test test device object Manage pressurizing vessel；
Step 2, coal sample physical compression
Apply certain pressure to coal sample by raw coal stress loading part, simulate reset pressure state locating for coal seam, applies Reset pressure controlled by oil pressure pump and pressure controller, it is 20MPa that maximum, which applies pressure,；
Step 3, environment harmful gas injects
Environment harmful gas mainly includes CO, CO2、H2S gas simulates down-hole coal bed atmosphere environment, according to down-hole coal bed Real gas atmosphere environment injects a certain amount of environmental gas to device inside coal body；
Step 4, coal sample gas pressure
High gas layer gas pressure, first are simulated by the methane gas that certain pressure is injected to coal sample in methane gas injection part It is 10MPa that alkane gas, which injects part highest to apply gas pressure,；
Step 5, environment temperature controls
By environment temperature control system, water bath with thermostatic control control is carried out to temperature locating for the experimental provision external world, makes experimental provision Coal body locating for coal body is in temperature constant state, and the highest environment temperature which is applied is 50 DEG C；
Step 6, methane oxidation bacterium solution is injected
The methane oxidation bacterium solution of culture is injected into experimental provision coal body by methane oxidation bacterium solution injection pipe, from upper port Injection, is dispersed in methane oxidation bacterium solution in coal body；
Step 7, environmental pH is adjusted
By pH regulating device to adjust down-hole coal bed pH value, adjusts it and meet coal seam actual environment pH；
The step 3, step 4, step 5, step 6, step 7 use control variate method, by change atmosphere, pH, pressure, The influence of temperature, methane-oxidizing bacteria night injection quantity research to methane oxidation flora degradation gas ability, analyzes factor variation pair The affecting laws of methane-oxidizing bacteria breeding and degradation gas ability；
Step 8, methane gas component is monitored on-line
Part is monitored on-line by methane gas, by the concentration of methane gas in Methane monitoring instrument on-line real time monitoring coal body, Lead to the changing rule of methane concentration to analyze methane-oxidizing bacteria resolution gas；
Step 9, environmental gas component monitors
By taking tracheae to connect Outside Vapor chromatograph in coal body, environmental interior gas component, concentration changing rule are analyzed, especially It is CO2Concentration variation, to study methane oxidizing bacteria growth process gas release rule；
Step 10, environmental liquids component analysis
By extractor coal body internal solution, coal body internal liquid phase change of component, analysis are analyzed using liquid phase mass spectrometric analysis instrument The changing rule of methane oxidizing bacteria growth process mesostate；By observing under microscope extracting solution, first is analyzed The growth and breeding situation of alkoxide bacterium.
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