CN101921811A - Method for culturing microalgae - Google Patents
Method for culturing microalgae Download PDFInfo
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- CN101921811A CN101921811A CN 201010231006 CN201010231006A CN101921811A CN 101921811 A CN101921811 A CN 101921811A CN 201010231006 CN201010231006 CN 201010231006 CN 201010231006 A CN201010231006 A CN 201010231006A CN 101921811 A CN101921811 A CN 101921811A
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Abstract
The invention relates to a method for culturing microalgae, which is characterized by comprising the following steps of: (1) fermenting to produce acid, enabling the supernatant which is acid liquor obtained after fermentation to flow to a culture device, and collecting the produced gas mixture in a gas storage tank; (2) inoculating the cultured microalgae to the acid liquor contained in the culture device for heterotrophic culture; and (3) circularly introducing the gas mixture in the gas storage tank to a photobioreactor for aerobic autotrophic culture. In the invention, the acid production stage through methane fermentation and the microalgae culturing process are coupled, and heterotrophy and autotrophy are combined; thus, oil-rich microalgae with high biomass is cultured, and meanwhile, a hydrogen-rich gas with high concentration can be obtained. The method is mainly used for culturing the oil-rich microalgae.
Description
Technical field
The present invention relates to little algae culture technique.
Background technology
Along with the continuous aggravation of global energy crisis, and the continuous enhancing of people's environmental consciousness, renewable resourcess such as biofuel are subjected to people and more and more pay close attention to.Biofuel is a kind of fatty acid monoester that is obtained through transesterification reaction by animals and plants or microbial oil and low-carbon alcohol, it is renewable, biodegradable, the burning and exhausting performance is good than fossil diesel fuel, is a kind of environmentally friendly energy, becomes the substitute of fossil diesel fuel gradually.Little algae is a kind of one-celled plants, has photosynthetic efficiency height, adaptive capacity to environment is strong, growth cycle is short and biomass is high characteristics.In recent years because the development of biofuel industry; the comprehensive exploitation of the oil-rich microalgae that fat content is higher and utilization have caused that people more and more pay close attention to; microalgae grease can solve present biodiesel raw material source and lack and problem of unstable; each country and geographic scientific research institution have carried out the research work of little algae cultivation aspect in succession, make its become gradually many countries increase income energy-conservation, turn bane into boon and protect the important means of environment.
In many characteristics of little algae, its stronger adaptive faculty has a wide range of applications.At present existing research work shows, little algae can grow in sanitary sewage or organic waste water, as patent CN101525576A, a kind of method of utilizing sanitary sewage and trade effluent to produce little algae is characterized in that, this method may further comprise the steps: the initial-stage culture step, after will handling also filtering sanitary sewage adding bio-reactor through one-level, add the algae kind in bio-reactor, stir culture under the condition of illumination, blowing air and carbonic acid gas grows into stationary phase up to microalgae cell; Grease is induced step, according to the industrial effluent after 1: 200～1: 1000 the volume ratio adding sugarcane bagasse fermentation, induces the oil and fat accumulation in the microalgae cell in bio-reactor; Extraction step, oil and fat accumulation reach stable after, extract the microalgae cell in the bio-reactor.Organic sewage still contains nutritive elements such as a large amount of nitrogen, phosphorus after handling through anaerobic-aerobic, wherein feed stack gas after little algae can utilize CO
2Carry out growth and breeding and accumulate grease, this had both reduced the cultivation cost of little algae, simultaneously sewage was carried out a biological disposal upon, and made it can recycle.
Little algae can also utilize biogas fermentation waste liquid-natural pond liquid to cultivate, as patent CN 101285075A, the coupling process that a kind of biogas fermentation and autotrophic freshwater microalgae are cultivated is disclosed, biogas fermentation and the photosynthetic cultivation of autotrophic freshwater microalgae are carried out simultaneously, it is characterized in that: 1) biogas fermentation: fermentation obtains biogas and natural pond liquid; 2) processing of natural pond liquid: the natural pond liquid that obtains behind the biogas fermentation of learning from else's experience, filter and remove remaining solid particulate, be 25%～100% of original content with the water dilution, as the full nutritional medium of autotrophic freshwater microalgae; 3) introducing of the inoculation of autotrophic freshwater microalgae and biogas: the autotrophic freshwater microalgae of packing into of the natural pond liquid after will handling is cultivated the bioreactor of usefulness, inserts the fresh autotrophic freshwater microalgae algae kind that spreads cultivation, and biogas is introduced reactor from reactor bottom; 4) autotrophic freshwater microalgae is cultivated and controlled: it is 1000Lx～2000Lx that autotrophic freshwater microalgae is cultivated early stage intensity of illumination, increases light intensity to 4000Lx～8000Lx, intermittence or 24h continuous illumination to cultivating the middle and later periods; 5) results of autotrophic freshwater microalgae cell: after the frustule cessation of growth cessation, stop the illumination ventilation, behind the collection frustule, remove the moisture in the frustule.Little algae is inoculated in the dilution natural pond liquid to a certain degree, and with the CO in the biogas
2For inorganic carbon source is carried out the autotrophy process, reach 6g/L through final biomass after 9 days, the coupling of little algae and biogas fermentation had both realized the cultivation of little algae in this process, improved the purity of biogas, reduce the environmental pollution that the discharging of natural pond liquid brings again, reached the purpose that increases society and economic benefit.But shortcomings such as aforesaid method mainly is the training method that adopts autotrophy, and it is slow to exist the speed of growth, and biomass is relatively low.
Summary of the invention
Technical problem to be solved by this invention is: product acid phase and little algae culturing process of coupling biogas fermentation, heterotrophism combines with autotrophy, can obtain the higher hydrogen-rich gas of concentration in the oil-rich microalgae of cultivating high-biomass.
For achieving the above object, the technical solution used in the present invention is:
(1) acidogenic fermentation
With animal excrement, agricultural crop straw, city domestic sewage in rural areas by using, industrial and agricultural production organic waste water and waste residue etc. is fermentation substrate, be placed on the fermentation reaction device, controlled temperature is at 25 ± 1 ℃ after adding active sludge, the 12-72h that ferments, and the gas mixture of generation is collected in the gas-holder.Described active sludge is meant the microorganism species that is used for the sewage disposal anaerobic ferment process.
(2) little algae is cultivated
The freshwater microalgae that the present invention cultivates comprises the autotrophic freshwater microalgae after wild environments such as coming from fresh water or the domestication, can be in each big algae kind preservation center or commercially available acquisition, as chlorella, grid algae and spirulina etc.Described little algae algae kind is chlorella (chlorella vulgaris), once be recorded among the laid-open U.S. Patents ublic specification of application US 20090288223 November 19 in 2009, also can obtain in Texas ,Usa algae kind preservation center, its deposit number is UTEX 2714.
The cultivation of algae kind: medium component is NH
4Cl 400mg/L, KH
2PO
456mg/L, K
2HPO
4108mg/L, MgSO
47H
2O 100mg/L, Glacial acetic acid 1mg/L, micro-1ml/L, the algae kind is inoculated in the substratum of the bacterium of going out through aseptic technique, places illumination box to leave standstill cultivation then, shake 3-4 every day, culture temperature is 15-30 ℃, and intensity of illumination is 2000-4000Lux, cultivates 7 days.
Acid solution is cultivated: acidogenic fermentation finishes after the acid solution of screw extrusion press after separating flow to culture apparatus, inoculates to carry out heterotrophism behind little algae and cultivate.Culture condition is temperature 15-30 ℃, cultivates 5 days.
Aerated culture: after treating that organic carbon source exhausts, carry out autotrophy in gas mixture (main component is hydrogen and carbonic acid gas) the feeding photo-biological reactor with the fermentation generation and cultivate, culture condition is temperature 15-30 ℃, and intensity of illumination is 4000-8000Lux, cultivates 7 days.
Marsh gas fermentation processes roughly can be divided into two stages, and the firstth, produce acid, being about to organic matter degradation is micromolecular VFA such as acetate, propionic acid etc., produces hydrogen simultaneously, CO
2And minor N H
3, H
2Gases such as S; The secondth, produce methane, the i.e. main component of biogas.Because the acidogenic fermentation time is short, several hrs was by several days, and condition is easier to control, and the gas that produces mainly is hydrogen and carbonic acid gas, and produced methane process condition harshness, and the cycle is longer, generally needs about two months, and misoperation causes the failure of fermentation easily.Little algae not only utilizes organic carbon source such as acetate to grow fast, can also utilize carbonic acid gas in the gas to carry out autotrophy as inorganic carbon source and cultivate.Therefore acid process and little algae two stages of cultivation are combined, compare with biogas fermentation, improved the stability of total system, fermentation time also shortens greatly.To produce acid (hydrogen) fermentation and combine, have following beneficial effect with little algae cultivation:
(1) acidogenic fermentation time weak point only needs a few hours to several days, and little algae heterotrophism process growth velocity height, and the cycle is short, and this system compares with whole biogas fermentation, and cycle and stability all are greatly improved.
(2) the autotrophy process of little algae has improved the concentration of hydrogen, for next step application such as fuel cell etc. provides good condition, and has further increased the biomass of little algae.
(3) this process turns waste into wealth, and has reduced environmental pollution, has realized the recycling of waste.
Description of drawings
Fig. 1 is a process route view of the present invention.
Embodiment
Embodiment 1: step 1: acidogenic fermentation.With 500g cow dung is fermentation substrate, adding water adjusting solids content is 6%, fill in the reaction unit, begin fermentation after inoculating 25% active sludge (taking from sewage work), temperature is controlled at 25 ± 1 ℃, the collection and confinement of gases that produces behind the fermentation 48h, carries out solid-liquid separation with tunning through screw extrusion press in gas-holder.
Step 2: the cultivation of algae kind: the algae kind of employing is chlorella vulgaris (available from Texas ,Usa algae kind preservation center, UTEX 2714).Medium component is NH
4Cl 400mg/L, KH
2PO
456mg/L, K
2HPO
4108mg/L, MgSO
47H
2O 100mg/L, Glacial acetic acid 1mg/L, micro-1ml/L, total fat content can reach more than 40% of dry cell weight in this substratum, is oil-rich microalgae.This algae experiment confirm can survive in acidogenic fermentation gained liquid.Concrete operations are: substratum is joined respectively in the Erlenmeyer flask (the ventilative film that seals of tool), sterilize after aseptic technique inserts the algae kind through 121 ℃, place illumination box to leave standstill cultivation then, shake every day 3 times, culture temperature is 28 ℃, intensity of illumination is 4000Lux (fluorescent lamp), cultivates 7 days.
Acid solution is cultivated: the supernatant liquor of getting after the solid-liquid separation is that acid solution is to the fermentation culture device, according to 10% inoculum size (algae kind OD
680Be 0.8) cultivate after inserting above-mentioned cultivation gained algae kind.Culture condition is 28 ℃ of temperature, and aeration-agitation cultivates that the sampling and measuring biomass is 15g/L after 5 days, and fat content is 38%.
Embodiment 2: present embodiment and embodiment 1 something in common repeat no more, difference is can proceed the autotrophy process after the heterotrophism cultivation, on the basis of embodiment 1, little algae after the heterotrophism cultivation is placed the pot type bioreactor, and the gas mixture that circulation feeds in the gas cylinder carries out the autotrophy cultivation, and culture condition is 28 ℃ of temperature, intensity of illumination is 6000Lux, measure micro algae biomass 18g/L after 7 days, fat content is 37%, and hydrogen purity is more than 90%.
Embodiment 3: step 1: acidogenic fermentation.With the 500g maize straw is fermentation substrate, with fill in the reaction unit after 500g cow dung is mixed, adding water adjusting solids content is 8%, begin fermentation after inoculating 25% active sludge (taking from methane-generating pit), temperature is controlled at 25 ± 1 ℃, the collection and confinement of gases that produces behind the fermentation 72h, carries out solid-liquid separation with tunning through screw extrusion press in gas-holder.All the other steps are with step 2 among the embodiment, and the sampling and measuring biomass is 18g/L after 5 days, and fat content is 39%.
Embodiment 4: step 1: acidogenic fermentation.With the 2L beer brewery water is that fermentation substrate fills in the reaction unit, begin fermentation after inoculating 20% active sludge (taking from methane-generating pit), temperature is controlled at 25 ± 1 ℃, and the collection and confinement of gases of generation is in gas-holder, behind the fermentation 12h, tunning is carried out solid-liquid separation through screw extrusion press.All the other are with the step 2 among the embodiment 1, and the sampling and measuring biomass is 10g/L after 5 days, and fat content is 35%.
Claims (6)
1. a little algae cultured method is characterized in that it comprises the steps: (1) acidogenic fermentation, and the supernatant liquor after the fermentation ends is that acid solution flow to culture apparatus, and the gas mixture of generation is collected in the gas-holder; (2) will carry out the heterotrophism cultivation through the acid solution that cultured microalgae is inoculated in the culture apparatus; (3) gas mixture circulation in the gas-holder being passed into the autotrophy of ventilating in the photo-biological reactor cultivates.
2. little algae cultured method according to claim 1, it is characterized in that described acidogenic fermentation is meant with animal excrement, agricultural crop straw, city domestic sewage in rural areas by using, industrial and agricultural production organic waste water and waste residue etc. are fermentation substrate, it is mixed with an amount of water place the fermentation reaction device, controlled temperature is at 25 ± 1 ℃ after adding active sludge, the 6-72h that ferments, the supernatant liquor of fermentation ends is that acid solution flow to biological reaction apparatus, the collection and confinement of gases of generation is in gas-holder.
3. little algae cultured method according to claim 2 is characterized in that described active sludge is meant the microorganism species that is used for the sewage disposal anaerobic ferment process.
4. little algae cultured method according to claim 1 is characterized in that the cultivation of described little algae is meant that medium component is NH
4Cl 400mg/L, KH
2PO
456mg/L, K
2HPO
4108mg/L, MgSO
47H
2O 100mg/L, Glacial acetic acid 1mg/L, micro-1ml/L, the algae kind is inoculated in the substratum of the bacterium of going out through aseptic technique, places illumination box to leave standstill cultivation then, shake 3-4 every day, culture temperature is 15-30 ℃, and intensity of illumination is 2000-4000Lux, cultivates 7 days.
5. little algae cultured method according to claim 1, it is characterized in that carrying out behind the little algae of described inoculation heterotrophism and cultivate and be meant: culture condition is temperature 15-30 ℃, and aeration-agitation was cultivated 5 days.
6. little algae cultured method according to claim 1 is characterized in that described aerated culture: the gas mixture that feeds the fermentation generation to bioreactor carries out the autotrophy cultivation, and culture condition is temperature 15-30 ℃, and intensity of illumination is 4000-8000Lux, cultivates 7 days.
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| CN102154110A (en) * | 2011-01-27 | 2011-08-17 | 华东理工大学 | High-yield microalgae cultivating method |
| WO2012065545A1 (en) * | 2010-11-16 | 2012-05-24 | 华东理工大学 | Microalgae culturing method for oil and lutein rapid accumulation |
| CN102618585A (en) * | 2012-03-28 | 2012-08-01 | 上海师范大学 | Method for producing hydrogen according to anaerobic fermentation of bloom-forming cyanobacteria |
| CN102732425A (en) * | 2012-06-06 | 2012-10-17 | 中南民族大学 | Method for producing microalgae through utilizing livestock and poultry excrement primary wastewater |
| CN102757981A (en) * | 2012-08-08 | 2012-10-31 | 山东省科学院能源研究所 | Method for preparing methane through algae residue anaerobic digestion |
| CN103194395A (en) * | 2013-04-22 | 2013-07-10 | 山东省科学院能源研究所 | Method for culturing microalgae by using biogas residues as raw materials |
| CN103589644A (en) * | 2013-11-28 | 2014-02-19 | 黑龙江省能源环境研究院 | Method for culturing oil-producing microalgae by using brewery sewage |
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| WO2015161577A1 (en) * | 2014-04-25 | 2015-10-29 | 厦门大学 | Membrane photobioreactor for treating nitrogen and phosphorus out of limits in biogas slurry and treating method therefor |
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| CN107012072A (en) * | 2017-04-10 | 2017-08-04 | 南昌大学 | A kind of photo-biological membrane reactor and the application in sewage disposal, carbon sequestration and microalgae recovery |
| CN108611276A (en) * | 2018-04-16 | 2018-10-02 | 仲恺农业工程学院 | A method of recycling treatment being carried out to distillery waste water using microalgae |
| CN111979155A (en) * | 2020-08-31 | 2020-11-24 | 江南大学 | Method for converting methane into single-cell protein by two-step method |
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-
2010
- 2010-07-20 CN CN 201010231006 patent/CN101921811B/en not_active Expired - Fee Related
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| WO2012065545A1 (en) * | 2010-11-16 | 2012-05-24 | 华东理工大学 | Microalgae culturing method for oil and lutein rapid accumulation |
| CN102154110B (en) * | 2011-01-27 | 2016-03-02 | 华东理工大学 | A kind of microalgae culture method of high yield |
| WO2012100583A1 (en) * | 2011-01-27 | 2012-08-02 | 华东理工大学 | Culturing method for microalgae with high yields |
| CN102154110A (en) * | 2011-01-27 | 2011-08-17 | 华东理工大学 | High-yield microalgae cultivating method |
| CN102618585A (en) * | 2012-03-28 | 2012-08-01 | 上海师范大学 | Method for producing hydrogen according to anaerobic fermentation of bloom-forming cyanobacteria |
| CN102732425B (en) * | 2012-06-06 | 2014-08-13 | 中南民族大学 | Method for producing microalgae through utilizing livestock and poultry excrement primary wastewater |
| CN102732425A (en) * | 2012-06-06 | 2012-10-17 | 中南民族大学 | Method for producing microalgae through utilizing livestock and poultry excrement primary wastewater |
| CN102757981A (en) * | 2012-08-08 | 2012-10-31 | 山东省科学院能源研究所 | Method for preparing methane through algae residue anaerobic digestion |
| CN103194395A (en) * | 2013-04-22 | 2013-07-10 | 山东省科学院能源研究所 | Method for culturing microalgae by using biogas residues as raw materials |
| CN103194395B (en) * | 2013-04-22 | 2014-06-04 | 山东省科学院能源研究所 | Method for culturing microalgae by using biogas residues as raw materials |
| CN103589644A (en) * | 2013-11-28 | 2014-02-19 | 黑龙江省能源环境研究院 | Method for culturing oil-producing microalgae by using brewery sewage |
| US10533148B2 (en) | 2014-04-25 | 2020-01-14 | Xiamen University | Membrane photobioreactor for treating nitrogen and phosphorus that are out of limits in biogas slurry and treating method thereof |
| WO2015161577A1 (en) * | 2014-04-25 | 2015-10-29 | 厦门大学 | Membrane photobioreactor for treating nitrogen and phosphorus out of limits in biogas slurry and treating method therefor |
| CN104789603A (en) * | 2015-05-19 | 2015-07-22 | 哈尔滨工业大学 | Method for treating starch wastewater and simultaneously generating renewable energy source |
| CN104789602A (en) * | 2015-05-19 | 2015-07-22 | 哈尔滨工业大学 | Method for treatment of starch wastewater and synchronous energy production by coculture of bacteria and microalgaes |
| CN105671095A (en) * | 2016-02-27 | 2016-06-15 | 昆明理工大学 | Cultivation method for promoting rapid accumulation of oil producing microalgae cells and grease |
| CN105802853A (en) * | 2016-05-20 | 2016-07-27 | 佛山市聚成生化技术研发有限公司 | Method for promoting porphyridium growth by virtue of aquatic product processing wastewater |
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| CN107012072B (en) * | 2017-04-10 | 2023-10-31 | 南昌大学 | Optical biomembrane reactor and application thereof in sewage treatment, carbon fixation and microalgae collection |
| CN108611276A (en) * | 2018-04-16 | 2018-10-02 | 仲恺农业工程学院 | A method of recycling treatment being carried out to distillery waste water using microalgae |
| CN111979155A (en) * | 2020-08-31 | 2020-11-24 | 江南大学 | Method for converting methane into single-cell protein by two-step method |
| CN113072177A (en) * | 2021-03-10 | 2021-07-06 | 南京理工大学 | Device and method for synchronously treating sludge digestive fluid and biogas by using in-situ algae |
| CN113072177B (en) * | 2021-03-10 | 2022-12-09 | 南京理工大学 | Device and method for synchronously treating sludge digestive fluid and biogas by using in-situ algae |
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