CN101875962B - Method for screening high-efficiency methanogen floras by bioelectrochemical technology - Google Patents
Method for screening high-efficiency methanogen floras by bioelectrochemical technology Download PDFInfo
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- CN101875962B CN101875962B CN2009102379350A CN200910237935A CN101875962B CN 101875962 B CN101875962 B CN 101875962B CN 2009102379350 A CN2009102379350 A CN 2009102379350A CN 200910237935 A CN200910237935 A CN 200910237935A CN 101875962 B CN101875962 B CN 101875962B
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Abstract
The invention relates to a method for screening high-efficiency methanogen floras by bioelectrochemical technology. The method consists of two parts, namely an anaerobic part and an electrochemical part; and the high-efficiency methane-producing microbial floras are screened form residual activated sludge and anaerobic sludge or riverway sediment under the bioelectrochemical action. The method comprises the following steps of: putting inoculums in an anaerobic digester, adding two electrodes, namely a positive electrode and a negative electrode into the digester, and inputting 0.1 to 2.0 V DCcurrent to the two electrodes; supplementing several nutrients to the digester continuously or in batches, controlling the pH value of the fermentation liquor to between 5 and 9, and controlling the temperature to between 30 and 40 DEG C; and completing the screening process after about 30 to 40 days to obtain the high-efficiency methanogen floras which are mixed microbial floras, wherein the mixed microbial floras mainly comprise hydrogen-oxidizing methanogen. The method is suitable for culturing high-efficiency methanogen in the process of starting producing methane by anaerobic fermentation of organic waste (including organic solid waste and organic wastewater).
Description
Technical field
The invention belongs to technical field of waste water processing, particularly a kind of technology and working method of screening methanogen.
Background technology
Anaerobic treatment is called anaerobic digestion again, is under oxygen free condition, by multiple mikrobe acting in conjunction, organism is decomposed and generation CH
4, CO
2, H
2O, H
2S and NH
3Process.Organic anaerobic treatment process can be divided into three phases: 1. hydrolysis, fermentation stage; After complicated organism is decomposed into micromolecular compound by the extracellular enzyme of mikrobe; Get in the cell of fermenting bacterial (being the acidifying bacterium); And be converted into simple more compound therein, simultaneously, bacterium utilizes the part material to synthesize new cellular material; 2. produce hydrogen and produce the acetate stage, voltaile fatty acid such as propionic acid, butyric acid and ethanol etc. are converted into acetate, H by one type of special bacterium (hydrogen-producing acetogenic bacteria)
2And CO
23. produce methane phase, utilize acetate, H by methanogenic bacterium
2And CO
2Produce methane (CH
4).
From phylogeny, methanogen can be divided into 5 orders, is respectively methagen order, methane coccus order, sarcina methanica order, methane germ order and methane ultrahigh-temperature Zoopagales.Methanogen is segmented again, can be divided into 19 genus: Methanobacterium, the thermophilic Pseudomonas of methane, the spherical Pseudomonas of methane, the avette Pseudomonas of methane, methane brevibacterium sp, methane germ genus, methane spirillum, methane grain Pseudomonas, methane bag shape Pseudomonas, methane Discomyces, methanococcus, methanogen genus, the leaf Pseudomonas of methane, methanothrix sp genus, Methanosarcina, methane leaf Pseudomonas, methane Halobacterium and methane salt Pseudomonas etc.The methanogen of isolation identification reaches kind more than 200 at present.Methanogen is the mikrobe of a type of extreme strictly anaerobic, chemoautotrophy or chemoheterotrophy, and this quasi-microorganism can utilize H
2+ CO
2, acetate, methyl compound (like methyl alcohol, methylamine, methyl sulfide, formate and methyl selenide) etc., and be translated into methane.
The biosynthesizing of methane and nitrogen fixedly be the unique metabolic process of methanogen.Find that at present the methane biosynthetic process has three kinds of approach in the methanogen: be the methane route of synthesis of raw material with acetate, with H
2With CO
2For the methane route of synthesis of raw material with the methyl compound is the methane route of synthesis of raw material.In the anaerobic treatment process, methanogen can utilize and produce the H that hydrogen produces the generation of acetate stage
2, CO
2With acetate etc., synthesizing methane.It is two kinds of main methane processes of producing in the anaerobic treatment process that acetic acid and methane producing and carbonic acid gas and hydrogen produce methane, respectively corresponding two types of methanogens: have a liking for the acetic acid and methane producing bacterium and have a liking for the producing hydrogen and methane bacterium.Because about 70% organism is converted into acetate through acidifying, hydrolysis meeting; And hydrogen generating quantity is less; The methane that causes producing in the anaerobic treatment process comes from the acetate conversion above 70%; Therefore have a liking for the acetic acid and methane producing bacterium and in the methanogen floras of traditional anaerobic digestion, preponderate, acetic acid and methane producing is the main path of methanation.Yet, in all methanogens of finding at present, only have 2 to belong to (eight folded methanococcus-Methanosarcina sp. and thread methanobacteria-Methanothrix sp.) for having a liking for the acetic acid and methane producing bacterium, and hydrolysis of organic matter, the other two kinds of product-H of acidifying
2And CO
2Can be utilized by other methanogen except that above-mentioned two genus methanogens, produce methane.Therefore, should in anaerobic treatment, screen and have a liking for the producing hydrogen and methane bacterium, and make it in the anaerobic treatment process, to preponderate, can change methanogen group structure in traditional anaerobic treatment process, change the approach that methane produces, quicken the generation of methane, improve the output of methane.
Summary of the invention
The object of the present invention is to provide that a kind of working cost is low, technology is simple, easy to operate, efficient methanogen screening method that energy consumption is low.The present invention is in order to overcome the deficiency that traditional anaerobic treatment exists, to utilize the bioelectrochemistry technology, through biological chemistry and electrochemical synergy, screen efficient methanogen, the generation of acceleration methane, and the output of raising methane.
Scheme of the present invention is following:
The mud that adds fermentor tank effective volume 1/3-1 in the anaerobic fermentation tank seals after logical nitrogen is got rid of fermenter system oxygen, and the temperature of fermentor tank is controlled at 30 ℃-40 ℃, and the pH value is controlled at 5.0-9.0, and stirring velocity is at 300-700rpm.In fermentor tank, add the liquid state organics material in the fermenting process, two battery lead plates that are fixed in the fermentor tank link to each other with direct supply through lead, and during fermentation, direct supply is in opened condition, and the voltage of direct supply output is 0.1-2.0V.Distance between the battery lead plate is 2-50 centimetre.Fermentation culture 20-30 days, can obtain containing the mixed solution of efficient methanogen.
In the fermenting process organic materials disposable, in fermentor tank, add in batches or continuously.
Organic materials comprises high concentrated organic wastewater, mud, rubbish from cooking, acetic acid or acetate solution etc.;
Be added with glucose, yeast extract paste, MgCl in the said organic materials
26H
2O or NaHCO
3
The anaerobic jar device structure is following among the present invention: the electrochemistry part is made up of adjustable direct supply, lead and two electrodes, is fixed with two electrode pads in the anaerobic fermentation tank, and two electrode pads are connected with adjustable direct supply through lead.Two electrode pads can be processed by conductive material, but said electro-conductive material is that iron, copper, aluminium, titanium, platinum, silver, gold or graphite etc. are nonmetal, and the electrode pad shape can be netted, block, strip, bar-shaped.
The output voltage range of adjustable direct supply described in the present invention is 0.1-2.0V, and preferable range is 0.4-1.2V.
The distance of said two electrode pads is at 5-30 centimetre.
The fermentor cultivation temperature is preferably 35～38 ℃ among the present invention; Cultivation pH is 6.5-7.5;
Pole plate area and pole plate distance are adjusted according to the fermentor tank size.
Reinforced, the continuously stirring operation in operation employing gap of fermentor tank.
Too fast in the fermenting process for preventing the acidifying in fermentor tank of liquid state organics material, can material effectively carried out preparatory acidifying under the control.Acidization is in advance: the method material handling that adopts the anaerobic hydrolysis-acidification bacterium.
The present invention screens efficient methanogen through the bioelectrochemistry effect from residual active sludge, anaerobic sludge or river bottom mud.
The charging of anaerobic fermentation tank can be adopted continous way or batch-type, and charging is adopted and contained the organic aqueous solution.
Inoculation uses mud to be active sludge, anaerobic sludge or river bottom mud.
The efficient methanogen that the present invention's screening obtains is a kind of mixed bacterial, and this mixed bacterial is main to have a liking for the producing hydrogen and methane bacterium.
After the volume of mixed solution reaches the fermentor tank effective volume in the fermentor tank, i.e. the 70%-75% of fermentor tank volume, screening process is accomplished, and obtains containing the mixture of efficient methanogen.Find that through Micro biological Tests the methanogen in this mixture is mainly has a liking for the producing hydrogen and methane bacterium, like Ka Liyasa methanogen (Methanogenium cariaci), cud methane tyrothricin (Methanobrevibacterruminantium) etc.The experiment of multiparity methane ability test finds that this mixture is than the high 0.25-0.85 of product methane ability times of common anaerobic sludge (comprising the anaerobic sludge that is used as inoculation originally).
Beneficial effect: invention characteristics of the present invention are through the lower voltage of input, adopt the bioelectrochemistry technology from from the bed mud in residual active sludge, anaerobic sludge or the river course of sewage work, filtering out the high-efficiency methanogen floras that differs from traditional anaerobic treatment advantage methanogen floras.Electrochemical adding can change bioprocess in traditional system for anaerobic treatment, mainly is that methane produces approach.Through the collaborative screening that can realize efficient methanogen of electrochemistry and biological action, the high 0.25-0.85 of product methane ability force rate tradition anaerobic treatment advantage methanogen of the methanogen floras that sieves doubly.
Description of drawings
Fig. 1 is an equipment synoptic diagram of the present invention
Embodiment
Below in conjunction with implementing the present invention is described further:
Embodiment 1
Fermentor tank is cylindrical, and its volume is that (d * h=46mm * 80mm), effectively working volume is 100L to 130L, and the length of electrode pad is 2/3 of fermentor tank virtual height, and width is 2/3 of a fermentor tank diameter.Distance between the two is 20 centimetres.Said plate material is an iron, and said two electrode shapes are netted.In the anaerobic digester of Beijing sewage work, obtain anaerobic sludge, anaerobic sludge is joined in the fermentor tank, the add-on of mud (being inoculum size) is about 1/2 of fermentor tank effective volume, and the concentration of mud is 10g/L.After the logical nitrogen of fermentor tank is got rid of oxygen wherein, sealing.For guaranteeing in the fermentor tank enough nutritive substances are arranged, timing even flow in fermentor tank adds market and buys alimentary acetic acid, and the flow acceleration of acetic acid is undertaken by table 1:
The flow acceleration of table 1, acetic acid
Through temperature controlling system the temperature of fermentor tank is remained on 37 ± 1 ℃, remain between the 6.8-7.2 with sodium hydroxide or the hydrochloric acid pH value with fermentor tank through the pH value control system, stirring velocity is 500rpm.Direct supply is imported the voltage of 1.0V continuously in the fermenting process to two electrodes.After 20 days, the volume of mixed solution reaches its effective volume in the fermentor tank, and screening process is accomplished, and obtains containing the mixture of high-efficiency methanogen floras.Find that through Micro biological Tests the methanogen in this mixture is mainly has a liking for the producing hydrogen and methane bacterium, like Ka Liyasa methanogen (Methanogenium cariaci), cud methane tyrothricin () such as (Methanobrevibacter ruminantium).The experiment of multiparity methane ability test finds that this mixture is higher 0.85 times than the product methane ability of common anaerobic sludge (comprising the anaerobic sludge that is used as inoculation originally).
Embodiment 2
Fermentor tank is cylindrical, and its volume is that (d * h=26mm * 50mm), its effective working volume is 20L to 26.5L.The length of electrode pad is 1/2 of fermentor tank virtual height, and width is 2/3 of a fermentor tank diameter.Distance between the two is 5 centimetres.But said electro-conductive material is a copper, and said two electrode shapes are bar-shaped.Obtain bottom mud from the bottom in river course, Beijing, mud is joined in the anaerobic fermentation tank, the add-on of anaerobic sludge (being inoculum size) is about 1/4 of fermentor tank effective volume., the logical nitrogen of fermentor tank seals after getting rid of oxygen wherein.Even flow totalling COD is about the synthetic organic waste water of 2000mg/L in fermentor tank, and the composition of organic waste water is following: glucose 16.2g/L, yeast extract paste 0.42g/L, MgCl
26H
2O 0.3g/L, NaH
2CO
32g/L.The flow acceleration of synthetic wastewater is undertaken by table 2:
The flow acceleration of table 2, synthetic wastewater
Through temperature controlling system the temperature of fermentor tank is remained on 35 ± 1 ℃, remain between 6.8～7.2 with sodium hydroxide or the hydrochloric acid pH value with fermentor tank through the pH value control system, stirring velocity is 500rpm.Direct supply is imported the voltage of 0.7V continuously to two electrodes.After 30 days, after the volume of mixed solution reached its effective volume in the fermentor tank, screening process was accomplished, and obtained containing the mixture of efficient methanogen.Adopting molecular biology to carry out microbiological analysis finds; Methanogen in this mixture is mainly has a liking for the producing hydrogen and methane bacterium, like cud methane tyrothricin (Methanobrevibacter ruminantium), Wan Shi methane coccus (Methanococcus vannielii), Bu Leisi methane bag shape bacterium (Methanoculleus bourgense) etc.The analysis of multiparity methane ability test finds that this mixture is higher 0.76 times than the product methane ability of common anaerobic sludge (comprising the anaerobic sludge that is used as inoculation originally).
Embodiment 3
Fermentor tank is cylindrical, and its volume is that (d * h=26mm * 50mm), its effective working volume is 20L to 26.5L.The length of electrode pad is 1/2 of fermentor tank virtual height, and width is 2/3 of a fermentor tank diameter.Distance between the two is 50 centimetres.Electrode pad is a strip, and said electrode pad material is an aluminium, and said two electrode shapes can be netted, block, strips, bar-shaped.In the aeration tank of Beijing sewage work, obtain active sludge; And mud joined in the fermentor tank; The add-on of mud (being inoculum size) reaches the fermentor tank effective volume, and the concentration of mud is for (total solid (TS) is 4～10g/L, and volatility solid substance (V) is 3～8g/L)., the logical nitrogen of fermentor tank seals after getting rid of oxygen wherein.Through temperature controlling system the temperature of fermentor tank is remained on 35 ± 1 ℃, remain between 6.8～7.2 with sodium hydroxide or the hydrochloric acid pH value with fermentor tank through the pH value control system, stirring velocity is 500rpm.Direct supply is imported the voltage of 0.5V continuously to two electrodes.After 24 days, screening process is accomplished, and obtains containing the mixture of efficient methanogen.Find through the Micro biological Tests analysis; Methanogen in this mixture is mainly has a liking for the producing hydrogen and methane bacterium, like cud methane tyrothricin (Methanobrevibacter ruminantium), Wan Shi methane coccus (Methanococcus vannielii), Ka Liyasa methanogen (Methanogenium cariaci) etc.The analysis of multiparity methane ability test finds that this mixture is higher 0.32 times than the product methane ability of common anaerobic sludge (comprising the anaerobic sludge that is used as inoculation originally).
Embodiment 4
Fermentor tank is cylindrical, and its volume is that (d * h=60mm * 142mm), its effective working volume is 300L to 400L.The length of electrode pad is 1/2 of fermentor tank virtual height, and width is 2/3 of a fermentor tank diameter.Distance between two electrode pads is 2 centimetres.Electrode pad is netted; In the aeration tank of Beijing sewage work, obtain active sludge, and mud is joined in the fermentor tank, the add-on of mud (being inoculum size) reaches the fermentor tank effective volume; The concentration of mud is for (TS is 4～10g/L, and VS is 3～8g/L)., the logical nitrogen of fermentor tank seals after getting rid of oxygen wherein.Through temperature controlling system the temperature of fermentor tank is remained on 35 ± 1 ℃, remain between 6.8～7.2 with sodium hydroxide or the hydrochloric acid pH value with fermentor tank through the pH value control system, stirring velocity is 500rpm.Direct supply is imported the voltage of 0.2V continuously to two electrodes.In culturing process, regularly the venting port from fermentor tank carries out exhaust, and carries out methane detection in the exhaust.After 24 days, screening process is accomplished, and obtains containing the mixture of efficient methanogen.Find through the Micro biological Tests analysis; Methanogen in this mixture is mainly has a liking for the producing hydrogen and methane bacterium, like cud methane tyrothricin (Methanobrevibacterruminantium), Wan Shi methane coccus (Methanococcus vannielii), Ka Liyasa methanogen (Methanogeniumcariaci) etc.The analysis of multiparity methane ability test finds that this mixture is higher 0.32 times than the product methane ability of common anaerobic sludge (comprising the anaerobic sludge that is used as inoculation originally).
Claims (7)
1. the method for bioelectrochemistry screening high-efficiency methanogen floras comprises the steps: in anaerobic fermentation tank, to add the mud of fermentor tank effective volume 1/3-1, and fermentation culture was carried out in sealing after logical nitrogen was got rid of fermenter system oxygen; In fermentor tank, add the liquid state organics material in the fermenting process; Two battery lead plates that are fixed in the fermentor tank link to each other with direct supply through lead, and during fermentation, direct supply is in opened condition; The direct supply output voltage is 0.1-2.0V; Distance between the battery lead plate is 2-50 centimetre, cultivates the mixed solution that promptly obtained containing efficient methanogen in 20-30 days, and said mud is active sludge.
2. according to the method for the said bioelectrochemistry screening of claim 1 high-efficiency methanogen floras, it is characterized in that said fermentation culture temperature is controlled at 30 ℃～40 ℃, the pH value is controlled at 5-9, and stirring velocity is at 300-700rpm.
3. according to the method for claim 1 or 2 said bioelectrochemistry screening high-efficiency methanogen floras, it is characterized in that said fermentation culture temperature is controlled at 35 ℃～38 ℃, the pH value is controlled at 6-8.
4. according to the method for the said bioelectrochemistry screening of claim 1 high-efficiency methanogen floras, it is characterized in that said organic materials comprises high concentrated organic wastewater, mud, rubbish from cooking, acetic acid or acetate solution.
5. according to the method for claim 1 or 4 said bioelectrochemistry screening high-efficiency methanogen floras, it is characterized in that being added with in the said organic materials glucose, yeast extract paste, MgCl
26H
2O or NaHCO
3
6. according to the method for claim 1 or 2 said bioelectrochemistry screening high-efficiency methanogen floras, it is characterized in that said adjustable direct supply is 0.4-1.2V to the voltage of two electrodes input.
7. according to the method for claim 1 or 2 said bioelectrochemistry screening high-efficiency methanogen floras, it is characterized in that said high-efficiency methanogen floras is a kind of mixed bacterial, this mixed bacterial is main to have a liking for the producing hydrogen and methane bacterium.
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| CN104404122B (en) * | 2014-11-04 | 2016-04-20 | 中国矿业大学 | Coal seam methanogen floras current active and colony balance degree feature method for rapidly judging |
| CN108265081A (en) * | 2018-01-24 | 2018-07-10 | 中国石油大学(北京) | A kind of simple method for producing high quality biological methane |
| CN109355314B (en) * | 2018-11-14 | 2022-02-01 | 郑鲁生 | Method for producing combustible gas by using kitchen garbage and sludge |
| CN110498587A (en) * | 2019-08-23 | 2019-11-26 | 昆明理工大学 | A kind of method of electrochemical couple anaerobic fermentation processing milk cattle cultivating waste |
| CN111458261A (en) * | 2020-04-28 | 2020-07-28 | 常州工程职业技术学院 | Construction method of system for determining methane production potential of straws |
| CN114891676A (en) * | 2022-05-13 | 2022-08-12 | 淮阴工学院 | High-performance microorganism preparation method for promoting municipal organic waste co-treatment |
| CN114958696A (en) * | 2022-07-19 | 2022-08-30 | 中国科学院生态环境研究中心 | Methanogen culture medium and culture method |
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