CN106754492A - Microorganism species preparation method and its application of biogas are prepared for fermenting enteromorpha - Google Patents
Microorganism species preparation method and its application of biogas are prepared for fermenting enteromorpha Download PDFInfo
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- CN106754492A CN106754492A CN201611123506.7A CN201611123506A CN106754492A CN 106754492 A CN106754492 A CN 106754492A CN 201611123506 A CN201611123506 A CN 201611123506A CN 106754492 A CN106754492 A CN 106754492A
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- enteromorpha
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/02—Separating microorganisms from their culture media
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P5/00—Preparation of hydrocarbons or halogenated hydrocarbons
- C12P5/02—Preparation of hydrocarbons or halogenated hydrocarbons acyclic
- C12P5/023—Methane
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/30—Fuel from waste, e.g. synthetic alcohol or diesel
Abstract
The invention discloses a kind of salt-durable microbe flora preparation method and its application that biogas is prepared for fermenting enteromorpha,, with Enteromorpha as matrix, using the silt and seawater collected on Tideland resources, through continuously fermenting, domestication obtains the microorganism species for it.The flora that the present invention is obtained can keep good active and higher gas production in salinity is for 10 ‰ ~ 20 ‰ seawater, and Enteromorpha of being degraded in the system that salinity is less than 35 ‰ forms methane, can realize under conditions of not using fresh water to clean Enteromorpha original material, realize the efficient degradation to Enteromorpha, reach the purpose of saving water resource and time, and biological flue gas can be prepared while the harm that Enteromorpha is caused to coastal ocean environment is reduced, with good economic benefit and environmental benefit.
Description
Technical field
The invention belongs to microorganism and bioenergy technical field, and in particular to a kind of to prepare biogas for fermenting enteromorpha
Salt-durable microbe flora preparation method and its application.
Background technology
Enteromorpha（Enteromorpha）It is a kind of large-scale green alga of Chlorophyceae, Ulvaceae, frequent species includes intestines Enteromorpha, flat
Enteromorpha, edge pipe Enteromorpha, bar Enteromorpha etc..Enteromorpha has indomitable vitality, faster environmental suitability higher, reproduction speed,
Therefore, its a large amount of growths can cause very strong inhibitory action to the growth of other algae in same marine site.In recent years, due to ocean
Increasing environmental pollution, the eutrophication of seawater, result in the pernicious breeding of Enteromorpha.Meanwhile, Enteromorpha be floated to seashore rot it is smelly
High risks are not only caused to environment, and has threatened the development of inshore fishing and tourist industry.The outburst of green alga Enteromorpha causes
The extensive concern of society, but green alga Enteromorpha is containing about protein 9%-24%, fatty 0.9%-2%, crude fibre 8%-18%, can be as
Produce the raw material of biogas.
Biogas belongs to secondary energy sources, is one kind of regenerative resource, is that organic matter is produced by after microbiological anaerobic fermentation
Flammable mixed gas.The main component of biogas is methane（45~70%）And carbon dioxide（25~55%）, also contain
Other a small amount of gases, such as carbon monoxide, hydrogen sulfide, hydrogen, nitrogen.Biogas fermentation can reduce containing for organic matter in raw material
Amount, reduces environmental pollution.Biogas residue after fermentation ends can be circulated for producing organic feed and fertilizer again.The low heat value of biogas
It is 20-25MJ/m3, every cubic metre of biogas can produce 0.7 kilogram of anthracitic heat after burning completely.At present, countries in the world
Have begun to for biogas to be used as fuel and for illuminating, replace the effect of gasoline, diesel engine machine also fine with biogas.
In the prior art, it is no lack of the report that biogas is produced using microbial fermentation Enteromorpha.Such as《Different pretreatments and fermentation
Influence of the condition to Enteromorpha biogas yield》、《The shadow of granular size and several preprocess methods to Enteromorpha producing methane through anaerobic fermentation
Ring》Enteromorpha anaerobic fermentation production biogas is reported Deng article, as a result shows that Enteromorpha has good production potential.Patent CN
101418315A, CN 102876727A, CN 104642530A provide various methods for producing biogas as raw material with Enteromorpha, but
These methods need to use clear water to slough the salinity in Enteromorpha or prepare zymotic fluid with fresh water, and this is due to Enteromorpha Yu Haiyang
In algae, Enteromorpha after results carries a large amount of sea salt, and the fermentative microflora used in conventional technology is then typically from lake
It is enriched with sludge, hot spring mud, animal wastes and is obtained, it is not enough to sea salt tolerance, result in and produce natural pond using fermenting enteromorpha
Gas is difficult to industrialization production.It can be seen that, for the technique for solving current fermenting enteromorpha production biogas is unable to reach ideal effect
Problem, its problem demanding prompt solution is how to obtain one to have salt tolerant, efficient Enteromorpha degradation flora.
The content of the invention
It is an object of the invention to provide a kind of preparation method of the efficient, microorganism species of salt tolerant, and by the flora
Realize Enteromorpha degraded efficiently, stable and biogas production.
To achieve the above object, the present invention is adopted the following technical scheme that：
A kind of microorganism species preparation method that biogas is prepared for fermenting enteromorpha, comprises the following steps：
（1）Collect the black silt and seawater at Tideland resources earth's surface depth 5-10 cm；
（2）By collect silt and seawater press mass volume ratio 1-5 g/100 mL add anaerobic fermentation tank in, add do not clean
The Enteromorpha of salinity, Enteromorpha addition be 0.5-2 g/100 mL, then regulation system pH value be 7.0-7.5, salinity be 10 ‰-
20 ‰, sealing carries out flora domestication in 15-30 days after fermentation at 25 ~ 40 DEG C, and layer 2-4 sterile gauze mistake is used after fermentation ends
Filter, separation and fermentation liquid and biogas residue；
（3）To not clean in the Enteromorpha addition gained zymotic fluid of salinity, total solids content is 0.5-5% in making fermentation tank, and is pressed
Biogas residue is 1-3 g/100 mL addition gained biogas residues with the mass volume ratio of zymotic fluid, and regulation system pH value is 6.8-7.8, salinity
It is 10-20 ‰, in being fermented 15-30 days at 25 ~ 40 DEG C, is filtered using layer 2-4 sterile gauze after fermentation ends, again separation and fermentation
Liquid and biogas residue；
（4）Repeat step（3）After 3-5 times, gained filtrate is the bacterium solution of the ST.EP of flora containing salt-durable microbe.
Ancient bacterium in gained flora ST.EP is identified, as a result show, it includes following major microorganisms：Methane phase
Bag Pseudomonas（Methanofollis）, mane methane Pseudomonas（Methanosaeta）, methane Peziza（Methanoplanus）, first
Alkane capsule Pseudomonas（Methanoculleus）, Methanosarcina（Methanosarcina）, methanospirillum category
（Methanospirillum）, methane decomposite leaf Pseudomonas（Methanolacinia）, methane rope Pseudomonas（Methanolinea）, it is hot naked
Zygosaccharomyces（Thermogymnomonas）, methane food Methylobacillus（Methanomethylovorans）, methane thermal Bacillus
（Methanothermobacter）, methane phase Pseudomonas（Methanogenium）, methane germ category（Methanosphaerula）；
Bacterium in gained flora ST.EP is identified, as a result show, it includes following major microorganisms：Hot-bulb Pseudomonas
（Thermococcoides）, ocean robe Pseudomonas（Oceanotoga）, spiral Pseudomonas（Sphaerochaeta）,KosmotogaCategory,
Amino Bacillus（Aminobacterium）, amino zygosaccharomyces（Aminomonas）, syntrophism zygosaccharomyces
（Syntrophomonas）, sugared Zymobacterium（Saccharofermentans）, anaerobism rod Pseudomonas（Anaerobaculum）, shuttle
Pseudomonas（Clostridium sensu strict）, Desulfovibrio（Desulfovibrio）, mangrove Bacillus
（Mangroviflexus）And the unfiled bacterium of Bacteroidetes（Bacteroidetes）.
Gained microorganism species can be used for fermenting enteromorpha and prepare biogas；Its operating procedure is：Gained bacterium solution is added and is fermented
In tank, its addition is the 40-80% of fermenter volume, then to adding the Enteromorpha that does not clean salinity, its addition in fermentation tank
It is 1-10g/100mL, while regulation system salinity is between 10-35 ‰, system initial pH value is between 6.8-7.8；Will fermentation
Air discharge in tank carries out anaerobic fermentation, and fermentation temperature is 25-40 DEG C, and the batch fermentation time is 15-30 days, continous way fermentation
Fed again when should be less than the 20% of average daily gas production tolerance is produced daily；Maintenance pH value is between 6.5-8.0 in fermentation process.
The Enteromorpha includes fresh Enteromorpha and dries Enteromorpha, and kind includes intestines Enteromorpha（Enteromorpha intestinalis）, flat Enteromorpha（E. compressa）, edge pipe Enteromorpha（E. linza）, bar Enteromorpha（E. clathrat）In
One or more；Using it is preceding Enteromorpha is carried out into historrhexis with plug-in strip, pulverizer etc. after be more beneficial for fermentation carrying out.
Remarkable advantage of the invention is：
（1）The salt tolerance of gained flora of the invention is high, can simplify the pre-treatment step of fermenting enteromorpha, can reach the effect saved water and energy
Really；
（2）Flora preparation method of the present invention is simple, high through the salt-tolerant cultures stability and degradation efficiency after domestication；
（3）Show through prolonged application practice result, using microflora degradation Enteromorpha of the present invention, its average degradation rate is more than 75%, 15d
Accumulation produce methane quantity reach 250-300 mL/g, methane concentration is up to 55-65%.
Specific embodiment
In order that content of the present invention easily facilitates understanding, with reference to specific embodiment to of the present invention
Technical scheme is described further, but the present invention is not limited only to this.
A kind of microorganism species preparation method that biogas is prepared for fermenting enteromorpha, comprises the following steps：
（1）Collect Fujian Province Pingtan County dragon and phoenix head bathing beach intertidal zone（119.80 ° of E, 25.50 ° of N）Beach earth's surface depth 5-10
Black silt and seawater at cm；
（2）The silt collected and seawater are pressed into mass volume ratio 1-5:100（g/mL）In addition anaerobic fermentation tank, and add unclear
Used as matrix, Enteromorpha addition is 0.5-2 g/100 mL to the Enteromorpha of the desalinization of soil by flooding or leaching point；Then regulation system pH value is 7.0-7.5, salt
It is 10 ‰ -20 ‰ to spend, sealing after at 25 ~ 40 DEG C fermentation carry out flora domestication within 15-30 days, after fermentation ends use layer 2-4 without
Bacterium filtered through gauze obtains zymotic fluid and biogas residue；
（3）To not clean in the Enteromorpha addition gained zymotic fluid of salinity, total solids content is 0.5-5% in making fermentation tank, and is pressed
Biogas residue is 1-3 with the mass volume ratio of zymotic fluid:100（g/mL）Gained biogas residue is added, regulation system pH value is 6.8-7.8, salt
It is 10-20 ‰ to spend, and in being fermented 15-30 days at 25 ~ 40 DEG C, is filtered using layer 2-4 sterile gauze after fermentation ends, and hair is separated again
Zymotic fluid and biogas residue；
（4）Repeat step（3）After 3-5 times, gained filtrate is the bacterium solution of flora ST.EP.
Ancient bacterium in gained flora ST.EP is identified, is as a result shown, its major microorganisms for including and abundance are such as
Under：Methane phase bag Pseudomonas（Methanofollis）（50.15%）, mane methane Pseudomonas（Methanosaeta）（24.58%）, first
Alkane Peziza（Methanoplanus）（17.59%）, methane capsule Pseudomonas（Methanoculleus）（2.19%）, methane eight folds ball
Pseudomonas（Methanosarcina）（1.35%）, methanospirillum category（Methanospirillum）（1.14%）, methane decomposite leaf Pseudomonas
（Methanolacinia）（0.55%）, methane rope Pseudomonas（Methanolinea）（0.44%）, hot naked zygosaccharomyces
（Thermogymnomonas）（0.42%）, methane food Methylobacillus（Methanomethylovorans）（0.24%）, methane thermal
Bacillus（Methanothermobacter）（0.07%）, methane phase Pseudomonas（Methanogenium）（0.03%）, methane germ
Category（Methanosphaerula）（0.01%）And 1.23% unfiled ancient bacterium；
Bacterium in gained flora ST.EP is identified, is as a result shown, its major microorganisms for including and abundance are as follows：Heat
Coccus（Thermococcoides）（9.59%）, ocean robe Pseudomonas（Oceanotoga）（8.42%）, spiral Pseudomonas
（Sphaerochaeta）（5.85%）,KosmotogaCategory（4.43%）, amino Bacillus（Aminobacterium）（3.49%）,
Amino zygosaccharomyces（Aminomonas）（3.12%）, syntrophism zygosaccharomyces（Syntrophomonas）（3.01%）, sugared zymophyte
Category（Saccharofermentans）（2.35%）, anaerobism rod Pseudomonas（Anaerobaculum）（2.31%）, fusobacterium
（Clostridium sensu strict）（2.08%）, Desulfovibrio（Desulfovibrio）（1.79%）, mangrove bacillus
Category（Mangroviflexus）（1.43%）And 32.91% unfiled bacterium, these unfiled bacteriums belong to Bacteroidetes
（Bacteroidetes）.
The Enteromorpha includes fresh Enteromorpha and dries Enteromorpha, and kind includes intestines Enteromorpha（E. intestinalis）, flat waterside
Tongue（E. compressa）, edge pipe Enteromorpha（E. linza）, bar Enteromorpha（E. clathrat）In one or more；Using preceding
The carrying out of fermentation is more beneficial for after Enteromorpha is carried out into historrhexis with plug-in strip, pulverizer etc..
In above-mentioned steps, the reagent of pH value is adjusted including in hydrochloric acid, calcium hydroxide, NaOH, sodium acid carbonate, sodium carbonate
One or more, detecting instrument uses acidometer.
In above-mentioned steps, the reagent for adjusting salinity includes fresh water, seawater, sodium chloride, potassium chloride, magnesium chloride, magnesium sulfate, sulphur
One kind or several in sour ammonium, sodium acid carbonate, calcium chloride, potassium chloride, potassium dihydrogen phosphate, sodium bromide, strontium chloride, ferric citrate
Kind, detecting instrument uses salinometer.
Embodiment 1
During gained bacterium solution added into fermentation tank, its addition is the 40% of fermenter volume, is not then cleaned to being added in fermentation tank
The Enteromorpha of salinity, its addition is 1g/100mL, while regulation system salinity is 10 ‰, system initial pH value is 6.8；Will fermentation
Air discharge in tank carries out anaerobic fermentation, and fermentation temperature is 25 DEG C, batch fermentation 30 days, maintains pH value to exist in fermentation process
Between 6.5-7.0.After fermentation ends, the degradation rate of Enteromorpha is 79%, and gas production is 263mL/g, and methane concentration is 59%.
Embodiment 2
During gained bacterium solution added into fermentation tank, its addition is the 80% of fermenter volume, is not then cleaned to being added in fermentation tank
The Enteromorpha of salinity, its addition is 10g/100mL, while regulation system salinity is 35 ‰, system initial pH value is 7.8；Will hair
Air discharge in fermentation tank carries out anaerobic fermentation, and fermentation temperature is 40 DEG C, batch fermentation 15 days, maintains pH value to exist in fermentation process
Between 7.5-8.0.After fermentation ends, the degradation rate of Enteromorpha is 75%, and gas production is 258 mL/g, and methane concentration is 56.5%.
Embodiment 3
During gained bacterium solution added into fermentation tank, its addition is the 60% of fermenter volume, is not then cleaned to being added in fermentation tank
The Enteromorpha of salinity, its addition is 5g/100mL, while regulation system salinity is 20 ‰, system initial pH value is 7.2；Will fermentation
Air discharge in tank carries out anaerobic fermentation, and fermentation temperature is 30 DEG C, and continous way fermentation should be less than average daily aerogenesis in daily output tolerance
Amount 20% when feed again；Maintenance pH value is between 6.8-7.5 in fermentation process.After fermentation ends, the degradation rate of Enteromorpha is
78%, gas production is 277 mL/g, and methane concentration is 62.5%.
The foregoing is only presently preferred embodiments of the present invention, all impartial changes done according to scope of the present invention patent with
Modification, should all belong to covering scope of the invention.
Claims (6)
1. a kind of microorganism species preparation method that biogas is prepared for fermenting enteromorpha, it is characterised in that comprise the following steps：
（1）Collect the black silt and seawater at Tideland resources earth's surface depth 5-10 cm；
（2）The silt and seawater that will be collected are added in anaerobic fermentation tank, and the Enteromorpha of salinity is not cleaned in addition, then regulation system pH
It is worth for 7.0-7.5, salinity are 10 ‰ -20 ‰, sealing carries out flora domestication, fermentation ends in 15-30 days after fermentation at 25 ~ 40 DEG C
Filtered using layer 2-4 sterile gauze afterwards, separation and fermentation liquid and biogas residue；
（3）To not clean in the Enteromorpha addition gained zymotic fluid of salinity, total solids content is 0.5-5% in making fermentation tank, and is added
Enter gained biogas residue, regulation system pH value is 6.8-7.8, salinity is 10-20 ‰, in being fermented 15-30 days at 25 ~ 40 DEG C, ferment knot
Filtered using layer 2-4 sterile gauze after beam, again separation and fermentation liquid and biogas residue；
（4）Repeat step（3）After 3-5 times, gained filtrate is the bacterium solution containing salt-durable microbe flora.
2. the microorganism species preparation method of biogas is prepared for fermenting enteromorpha according to claim 1, it is characterised in that
Step（2）Middle silt is 1-5 g/100 mL with the mass volume ratio of seawater；The amount for adding Enteromorpha is 0.5-2 g/100 mL.
3. the microorganism species preparation method of biogas is prepared for fermenting enteromorpha according to claim 1, it is characterised in that
Step（3）Middle addition biogas residue is 1-3 g/100 mL with the mass volume ratio of zymotic fluid.
4. the microorganism species preparation method of biogas is prepared for fermenting enteromorpha according to claim 1, it is characterised in that institute
Stating salt-durable microbe flora includes methane phase bag Pseudomonas, mane methane Pseudomonas, methane Peziza, methane capsule Pseudomonas, methane eight
Folded Coccus, methanospirillum category, methane decomposite leaf Pseudomonas, methane rope Pseudomonas, hot naked zygosaccharomyces, methane food Methylobacillus, methane
Hot rod Pseudomonas, methane phase Pseudomonas, the ancient bacterium of methane germ category.
5. the microorganism species preparation method of biogas is prepared for fermenting enteromorpha according to claim 1, it is characterised in that institute
Stating salt-durable microbe flora includes hot-bulb Pseudomonas, ocean robe Pseudomonas, spiral Pseudomonas, amino Bacillus, amino zygosaccharomyces, mutually
Battalion's zygosaccharomyces, sugared Zymobacterium, anaerobism rod Pseudomonas, fusobacterium, Desulfovibrio, mangrove Bacillus bacteria.
6. the microorganism species that a kind of method as claimed in claim 1 is obtained, it is characterised in that prepare natural pond for fermenting enteromorpha
Gas；
Its operating procedure is：By in gained bacterium solution addition fermentation tank, its addition is the 40-80% of fermenter volume, then to hair
The Enteromorpha for not cleaning salinity is added in fermentation tank, its addition be 1-10g/100mL, while regulation system salinity 10-35 ‰ it
Between, system initial pH value is between 6.8-7.8；Air discharge in fermentation tank is carried out into anaerobic fermentation, fermentation temperature is 25-40
DEG C, the batch fermentation time is 15-30 days, and continous way fermentation feeds again when should be less than the 20% of average daily gas production tolerance is produced daily；
Maintenance pH value is between 6.5-8.0 in fermentation process.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2019081149A (en) * | 2017-10-31 | 2019-05-30 | 国立大学法人東京工業大学 | Anaerobic treatment method of wastewater and microorganism preparation |
| CN114410542A (en) * | 2022-02-15 | 2022-04-29 | 青岛中尚奇生物科技有限公司 | Vibrio sky blue and application thereof in enteromorpha degradation |
| CN114480201A (en) * | 2022-02-15 | 2022-05-13 | 青岛中尚奇生物科技有限公司 | Vibrio natriegens capable of strongly degrading enteromorpha protein and application of vibrio natriegens in preparation of organic fertilizer |
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2016
- 2016-12-08 CN CN201611123506.7A patent/CN106754492A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2019081149A (en) * | 2017-10-31 | 2019-05-30 | 国立大学法人東京工業大学 | Anaerobic treatment method of wastewater and microorganism preparation |
| CN114410542A (en) * | 2022-02-15 | 2022-04-29 | 青岛中尚奇生物科技有限公司 | Vibrio sky blue and application thereof in enteromorpha degradation |
| CN114480201A (en) * | 2022-02-15 | 2022-05-13 | 青岛中尚奇生物科技有限公司 | Vibrio natriegens capable of strongly degrading enteromorpha protein and application of vibrio natriegens in preparation of organic fertilizer |
| CN114410542B (en) * | 2022-02-15 | 2023-08-11 | 青岛中尚奇生物科技有限公司 | Vibrio paradise and application thereof in degradation of enteromorpha |
| CN114480201B (en) * | 2022-02-15 | 2023-08-11 | 青岛中尚奇生物科技有限公司 | Vibrio natriurus capable of strongly degrading enteromorpha protein and application of vibrio natriurus in preparation of organic fertilizer |
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Application publication date: 20170531 |