CN105713935B - A kind of method of microalgae mixed culture production grease - Google Patents
A kind of method of microalgae mixed culture production grease Download PDFInfo
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/20—Air quality improvement or preservation, e.g. vehicle emission control or emission reduction by using catalytic converters
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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
- Y02E50/10—Biofuels, e.g. bio-diesel
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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
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/59—Biological synthesis; Biological purification
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Abstract
The invention discloses a kind of methods that microalgae is mixed production grease, including following content: (1) micro-algae culture medium and scenedesmus obliquus FSH-Y2 seed liquor being added in bioreactor, the pH value for adjusting cultivating system is 10~12, is passed through CO in gas2Content control in 5v% hereinafter, culture 2~5 days；(2) pH value for adjusting cultivating system is 8~10, and access single needle algae SS-B1 seed liquor is mixed, and is passed through CO in gas2Content is 5v%~45v%；(3) culture to growth stationary phase terminates, harvesting microalgae cell；The scenedesmus obliquus (Scenedesmus obliqnus) FSH-Y2 and single needle algae (Monoraphidium sp.) SS-B1, it respectively at September in 2012 11 days and is preserved in " China Committee for Culture Collection of Microorganisms's common micro-organisms center " on April 15th, 2013, deposit number is respectively CGMCC No.6551 and CGMCC No.7479.The method of the present invention improves microdisk electrode system to high concentration CO2Tolerance and dissolubility, improve carbon sequestration efficiency, the harvest yield of microalgae grease significantly improves, and is able to carry out the production of biodiesel.
Description
Technical field
The invention belongs to biotechnologys and field of biological energy source, and in particular to one kind is resistant to high concentration CO2And in alkali
Property environment under microalgae mixed culture production grease method.
Background technique
Due to the increase of fossil energy reduced increasingly and cause greenhouse effects using fossil energy, more and more scientific researches
Worker focuses on sight in the development and utilization of renewable energy.Biomass energy is as renewable energy most important on the earth
Source, it includes forestry biomass, crops, water plant, agricultural wastes etc..In many biomass energies, microalgae is
Important renewable resource.They are with widely distributed, biomass is big, photosynthetic efficiency is high, strong environmental adaptability, growth
The features such as period is short, biomass yield is high.Containing unique primary or secondary metabolite, complex chemical composition in its cell.It is micro-
The solar conversion efficiency of algae can reach 3.5%, be the potential resource for producing drug, fine chemical product and New-type fuel, from microalgae
Obtained in fatty acid can be converted to Fatty acid methyl ester, i.e. biodiesel.
With the development of world economy, a large amount of fossil energy using and consuming, and leads to the shortage of the energy and environment
Worsening, especially CO2Sharply increase caused by greenhouse effects it is increasingly severe, the growth cycle of microalgae is short, photosynthetic effect
Rate is high, CO2Fixed efficiency is high, up to 10 times or more of terrestrial plant under certain condition, can not only reduce CO2Discharge, simultaneously
Also reduce toxigenic capacity；Except CO2Outside, the ingredients such as some SOx, NOx in exhaust gas are cleaned place also with the metabolism of microalgae
Reason effectively reduces noxious gas emission, therefore is most possible at present using microalgae grease as the biodiesel that raw material produces
Meet the renewable energy of fuel needed for the world transports.
At present for the more of the oil-producing microalgaes such as chlorella, scenedesmus research.CN20110144545.6 discloses one plant of scenedesmus
The growth of algae strain, algae strain can be grown using artificial medium or appropriately processed waste water, its main feature is that lipid-producing is higher than
Most of at present to divide algae strain, algae strain application field includes CO2Fixation, the purification of waste water, grease, protein, pigment, shallow lake
The production of powder, polysaccharide, nucleic acid.CN20120154470.4 disclose the micro- quasi- ball algae of one plant of rich oil marine microalgae (Nannochloropsis gaditana ) algae strain and its application, the algae strain can in the environment of pH=4.5 normal growth, oil
Rouge content is up to 35%.CN20111019480.X disclose one plant of microalgae algae strain (Mychonases sp .) and its for producing
The application of biodiesel can produce the polyunsaturated fatty acid of high added value, including linolenic acid C18:3 and mind using algae strain
The byproduct of high added value is obtained while obtaining biodiesel through sour C24:1.These patents are all not directed to algae pair
The tolerance of carbon dioxide.CN102703326A discloses a kind of high CO2The microalgae and its selection of tolerance and fixed rate,
But the strain of algae provided by the patent is not directed to the fat content of algae strain.Above-mentioned patent or it cannot efficiently utilize CO2Production
Fat content is not high enough in grease or the biomass of acquisition.Especially in practical applications, as CO in environment2Volume fraction
When greater than 5v%, the growth of most of microalgae will be suppressed, carbon sequestration low efficiency；General microalgae is suitable in neutral conditions simultaneously
Growth is unfavorable for the growth of microalgae under conditions of slant acidity or meta-alkalescence, and microalgae utilizes CO2Usually to be dissolved in water
In HCO3 -Existing for ionic species, solubility is low in a neutral environment for carbon dioxide, is unfavorable for algae and is absorbed and utilized.
Liu Pinghuai etc. (organic carbon source grows single needle frustule, oil and fat accumulation and photosynthetic influence, bioengineering,
2012,33 (18): 224-246) a kind of production method using organic carbon source culture single needle algae is described, although culture terminates
Biomass has been more than 10g/L, but this kind of mode is single needle algae Heterotrophic culture mode, utilizes the organic carbons such as glucose in incubation
Source realizes that cell is grown, and this training method do not utilize CO2Etc. inorganic carbon source economy, and the addition of organic carbon source,
Incubation is also easy to produce microbiological contamination problem, influences the growth of frustule.
Summary of the invention
It is not able to satisfy tolerance for existing algae and absorbs high concentration CO2, fixed CO2The problem of low efficiency, the present invention provide
One kind is resistant to high concentration CO2And the method that microalgae mixed culture produces grease under alkaline environment.The method of the present invention mentions
High microdisk electrode system is to high concentration CO2Tolerance and dissolubility, improve carbon sequestration efficiency, the harvest yield of microalgae grease is bright
It is aobvious to improve, it is able to carry out the production of biodiesel.
The method of microalgae mixed culture production grease of the present invention, including following content: (1) the raw grid by micro-algae culture medium and tiltedly
Algae FSH-Y2 seed liquor is added in bioreactor, and the pH value for adjusting cultivating system is 10~12, is passed through CO in gas2's
Content control is 5% hereinafter, culture 2~5 days；(2) pH value for adjusting cultivating system is 8~10, accesses single needle algae SS-B1 seed
Liquid is mixed, and CO in gas is passed through2Content is 5v%~45v%；(3) culture to growth stationary phase terminates, harvesting microalgae
Cell；The scenedesmus obliquus (Scenedesmus obliqnus) FSH-Y2 and single needle algae (Monoraphidium sp) SS-
B1 respectively at September in 2012 11 days and is preserved in that " China Committee for Culture Collection of Microorganisms is common on April 15th, 2013
Microorganism " center ", deposit number are respectively CGMCC No. 6551 and CGMCC No. 7479.
Frustule is in spindle under the microscope for FSH-Y2 algae strain of the present invention, grows thickly, there is cell envelope package, color
For bottle green；Single algae cell diameter is about 6~10 μm.Titanium dioxide can be preferably absorbed and utilized in algae strain at a high ph
Carbon, fast-growth breeding.
Single needle algae SS-B1 algae strain of the present invention is a kind of green algate of fresh water, and frustule is that length is leaf, green, and algae is a length of
It is 10~20 μm, 2~4 μm wide, pigment is included, it is S-shaped, bottle green that plate algae, which falls form,.Algae strain is resistant to the CO of high concentration2
And SO2, can use containing CO2And SO2Exhaust gas or flue gas carry out illumination autophyting growth and obtain rich grease-contained biomass, carbon sequestration
It is high-efficient.
In the present invention, micro-algae culture medium cultivates the liquid training of microalgae using BG11, SE, BBM well known in the art etc.
Support base.Preparing for scenedesmus obliquus FSH-Y2 seed liquor is as follows: the pH of culture medium being adjusted to 10~12, is 20~30 in temperature
DEG C, periodicity of illumination is that for 24 hours, brightness time ratio is 14:10, and intensity of illumination is 2000~10000Lux, and shaken cultivation is raw to logarithm
For a long time.Volume ratio 1:20~1:5 of the scenedesmus obliquus FSH-Y2 seed liquor and culture medium that are added in bioreactor, is passed through gas
Body is carbon dioxide and nitrogen, wherein CO2Content control in 3v%~5v%.
In the present invention, preparing for single needle algae SS-B1 seed liquor is as follows: the pH of culture medium being adjusted to 7~9, is in temperature
20~30 DEG C, periodicity of illumination is that for 24 hours, brightness time ratio is 14:10, and intensity of illumination is 2000~10000Lux, and shaken cultivation is extremely
Logarithmic growth phase；Single needle algae SS-B1 seed liquor and the volume ratio of culture medium are 1:10~1:4.The culture that the present invention is mixed
Temperature is 20~30 DEG C, and periodicity of illumination is that for 24 hours, brightness time ratio is 14:10, and intensity of illumination is 2000~10000Lux.It is passed through
Gas is it is preferable to use exhaust gas or flue gas, wherein CO2Content is 10v%~45v%, SO2Content is 200 × 10-6~600 × 10-6(v/
V).Mixed culture to growth stationary phase terminates, and by the modes harvesting microalgae cell such as being centrifuged, settling, measures dry cell weight and oil
Rouge content, dry cell weight can reach 6g/L, and fat content has been more than the 45% of dry cell weight.
Compared with prior art, the present invention can bring it is following the utility model has the advantages that
1, the present invention accesses different microalgae seed liquors using two-step method and is mixed, and initial reaction stage access tolerance is high
The scenedesmus obliquus FSH-Y2 algae strain of pH is grown under alkaline condition, can inhibit the life of microdisk electrode initial stage miscellaneous bacteria and pest and disease damage
It is long, so that microalgae is in growth vigor；High pH culture is conducive to the dissolution for being passed through high concentration carbon dioxide simultaneously, makes carbon dioxide more
It is easy to be absorbed and utilized by microalgae, helps to improve the fixed efficiency of carbon dioxide；
2, in the incubation for obtaining biomass using carbon dioxide and illumination, microalgae mixed culture is not microalgae list
The simple combination solely cultivated, both algaes are worked in coordination, and are had higher carbon sequestration efficiency than single algae and are obtained more
Biomass and fat content；
3, co-culture system of the invention is resistant to the CO of high concentration2And SO2, can use the CO in exhaust gas2It carries out
Autophyting growth, fixed CO2, alleviate current industrial society's bring greenhouse effects and exhaust pollution problems.
Specific embodiment
Below by embodiment, invention is further described in detail.In the present invention, wt% is mass fraction, and v% is body
Fraction.
The preparation of 1 microalgae seed liquor of embodiment
Microdisk electrode uses BG11 culture medium, and culture medium prescription is as shown in Table 1 and Table 2.
1 BG11 culture medium of table
* in 2 table 1 of table A5+Co solution composition
BG11 fluid nutrient medium is prepared first, in accordance with Tables 1 and 2, the pH tune of the culture medium of scenedesmus obliquus FSH-Y2 will be cultivated
Section is 10, the pH for cultivating the culture medium of single needle algae SS-B1 is adjusted to 7.5, then by scenedesmus obliquus FSH-Y2 and single needle algae SS-
B1 is inoculated in respectively in above-mentioned culture medium.It is cultivated in constant temperature illumination shaking table, cultivation temperature is 25 DEG C, and periodicity of illumination is light for 24 hours
Dark time ratio is 14:10, and intensity of illumination 5000Lux, 120rpm shaken cultivation to logarithmic growth phase obtains scenedesmus obliquus FSH-
Y2 seed liquor and single needle algae SS-B1 seed liquor, above-mentioned seed liquor is saved backup under 15 DEG C of dim lights.
The preparation of 2 microalgae grease of embodiment
(1) in bioreactor, FSH-Y2 seed liquor and micro-algae culture medium prepared by embodiment 1, FSH-Y2 is added
Seed liquor and the volume ratio of culture medium are 1:10, and culture medium uses BG11 culture medium, and the pH value control of culture medium is 10, culture
Intensity of illumination is 5000Lux, and periodicity of illumination is that for 24 hours, brightness time ratio is 14:10, is passed through the gaseous mixture of nitrogen and carbon dioxide
Body, wherein carbon dioxide content is 5v%.
(2) after cultivating 2 days, single needle algae SS-B1 seed liquor prepared by access embodiment 1, the volume of seed liquor and culture medium
Than controlling for the pH of 1:10, culture medium 8, it is passed through the mixed gas of nitrogen and carbon dioxide, wherein CO2Content is 30v%.
(3) enter stationary phase after cultivating 7 days, terminate culture, microalgae cell is harvested by centrifugation, measure dry cell weight and grease contains
Amount measures algae dried bean noodles weight under the conditions of -60 DEG C after vacuum freeze drying to constant weight, calculate yield of biomass, and use n-hexane:
Ethyl acetate method measures total lipid content.Being detected dry cell weight can reach 7.4g/L, and fat content is the 46.7% of dry cell weight.
The preparation of 3 microalgae grease of embodiment
(1) in bioreactor, FSH-Y2 seed liquor and micro-algae culture medium prepared by embodiment 1, FSH-Y2 is added
Seed liquor and the volume ratio of culture medium are 1:5, and culture medium uses BG11 culture medium, and the pH value control of culture medium is 12, culture
Intensity of illumination is 5000Lux, and periodicity of illumination is that for 24 hours, brightness time ratio is 14:10, is passed through the gaseous mixture of nitrogen and carbon dioxide
Body, wherein carbon dioxide content is 5v%.
(2) after cultivating 3 days, single needle algae SS-B1 seed liquor prepared by access embodiment 1, the volume of seed liquor and culture medium
Than controlling for the pH of 1:10, culture medium 10, it is passed through the mixed gas of nitrogen and carbon dioxide, wherein CO2Content is 25v%.
(3) enter stationary phase after cultivating 7 days, terminate culture, microalgae cell is harvested by centrifugation, measure dry cell weight and grease contains
Amount measures algae dried bean noodles weight under the conditions of -60 DEG C after vacuum freeze drying to constant weight, calculate yield of biomass, and use n-hexane:
Ethyl acetate method measures total lipid content.Being detected dry cell weight can reach 7.1g/L, and fat content is the 46.1% of dry cell weight.
The preparation of 4 microalgae grease of embodiment
(1) in bioreactor, FSH-Y2 seed liquor and micro-algae culture medium prepared by embodiment 1, FSH-Y2 is added
Seed liquor and the volume ratio of culture medium are 1:20, and culture medium uses BG11 culture medium, and the pH value control of culture medium is 11, culture
Intensity of illumination is 5000Lux, and periodicity of illumination is that for 24 hours, brightness time ratio is 14:10, is passed through the gaseous mixture of nitrogen and carbon dioxide
Body, wherein carbon dioxide content is 4v%.
(2) after cultivating 4 days, single needle algae SS-B1 seed liquor prepared by access embodiment 1, the volume of seed liquor and culture medium
Than controlling for the pH of 1:5, culture medium 9, it is passed through the mixed gas of nitrogen and carbon dioxide, wherein CO2Content is 40v%.
(3) enter stationary phase after cultivating 8 days, terminate culture, microalgae cell is harvested by centrifugation, measure dry cell weight and grease contains
Amount measures algae dried bean noodles weight under the conditions of -60 DEG C after vacuum freeze drying to constant weight, calculate yield of biomass, and use n-hexane:
Ethyl acetate method measures total lipid content.Being detected dry cell weight can reach 7.2g/L, and fat content is the 46.5% of dry cell weight.
Embodiment 5 prepares microalgae grease using flue gas
Preparation condition is same as Example 2, the difference is that step (2) is passed through gas and is changed to containing SO2And CO2Cigarette
Gas, CO in flue gas2Content be 10v%~45v%, SO2Content is 200 × 10-6~600 × 10-6(v/v).
Inoculum concentration when scenedesmus obliquus FSH-Y2 seed liquor, single needle algae SS-B1 seed liquor are individually cultivated is 20%；Mixing training
When supporting, the inoculum concentration of scenedesmus obliquus FSH-Y2 seed liquor and single needle algae SS-B1 seed liquor is respectively 10%.Independent condition of culture with
The condition of mixed culture is identical, starts to maintain pH to be 10, after culture 3 days, be passed through containing SO2And CO2Flue gas, CO in flue gas2Contain
Amount is 10v%~45v%, SO2Content is 200 × 10-6~600 × 10-6(v/v), maintaining the pH of reaction system is 9.
Enter stationary phase after culture 8 days, terminates culture, frustule is collected by centrifugation, vacuum freeze drying under the conditions of -60 DEG C
Algae dried bean noodles weight is measured after to constant weight, calculate yield of biomass, and use n-hexane: ethyl acetate method measures total lipid content, as a result
As shown in table 3.
The culture effect of table 3 mixed culture and single algae
Seen from table 3, relative to single algae, two-step method mixed culture not only can be with the CO of enduring high-concentration2, and can be with
It is resistant to certain density SO2, obtain higher biomass and fat content.It can be seen that can use two-step method mixing training
It supports and flue gas prepares microalgae grease, that is, realize the production of grease, while exhaust gas can be purified.
The preparation of 1 microalgae grease of comparative example
Preparation condition is same as Example 2, the difference is that FSH-Y2 seed liquor and single needle algae SS-B1 seed liquor are being trained
Starting is supported to be added in reactor.Enter stationary phase after culture 7 days, terminate culture, microalgae cell is harvested by centrifugation, measures dry cell weight
And fat content, dry cell weight can reach 6.1g/L, fat content is the 45.6% of dry cell weight.
The preparation of 2 microalgae grease of comparative example
Preparation condition is same as Example 3, the difference is that FSH-Y2 seed liquor and single needle algae SS-B1 seed liquor are being trained
Starting is supported to be added in reactor.Enter stationary phase after culture 7 days, terminate culture, microalgae cell is harvested by centrifugation, measures dry cell weight
And fat content, dry cell weight can reach 5.9g/L, fat content is the 45.1% of dry cell weight.
Claims (8)
1. a kind of method of microalgae mixed culture production grease, it is characterised in that including following content: (1) by micro-algae culture medium with
Scenedesmus obliquus FSH-Y2 seed liquor is added in bioreactor, and the pH value for adjusting cultivating system is 10~12, is passed through gas
Middle CO2Content control in 5v% hereinafter, culture 2~5 days；(2) pH value for adjusting cultivating system is 8~10, accesses single needle algae
SS-B1 seed liquor is mixed, and CO in gas is passed through2Content is 5v%~45v%, and wherein v% is volume fraction；(3) it cultivates
Terminate to growth stationary phase, harvesting microalgae cell；The scenedesmus obliquus (Scenedesmus obliqnus) FSH-Y2 and single needle
Algae (Monoraphidium sp.) SS-B1, respectively at September in 2012 11 days and it is preserved in " the micro- life of China on April 15th, 2013
Object culture presevation administration committee common micro-organisms center ", deposit number is respectively CGMCC No. 6551 and CGMCC No.
7479。
2. according to the method for claim 1, it is characterised in that: the preparation side of step (1) scenedesmus obliquus FSH-Y2 seed liquor
Method is as follows: the pH of culture medium being adjusted to 10~12, is 20~30 DEG C in temperature, periodicity of illumination is for 24 hours that brightness time ratio is
14:10, intensity of illumination are 2000~10000Lux, shaken cultivation to logarithmic growth phase.
3. method according to claim 1 or 2, it is characterised in that: the oblique raw grid being added in step (1) bioreactor
Algae FSH-Y2 seed liquor and the volume ratio of culture medium are 1:20~1:5.
4. according to the method for claim 1, it is characterised in that: micro-algae culture medium uses BG11, SE or BBM Liquid Culture
Base.
5. according to the method for claim 1, it is characterised in that: step (1) is passed through gas as carbon dioxide and nitrogen, wherein
CO2Content control in 3v%~5v%, wherein v% is volume fraction.
6. according to the method for claim 1, it is characterised in that: the preparation method of step (2) single needle algae SS-B1 seed liquor is such as
Under: the pH of culture medium is adjusted to 7~9, is 20~30 DEG C in temperature, periodicity of illumination is that for 24 hours, brightness time ratio is 14:10, light
It is 2000~10000Lux, shaken cultivation to logarithmic growth phase according to intensity；The volume ratio of single needle algae SS-B1 seed liquor and culture medium
For 1:10~1:4.
7. according to the method for claim 1, it is characterised in that: the temperature of step (2) mixed culture is 20~30 DEG C, illumination
Period is that for 24 hours, brightness time ratio is 14:10, and intensity of illumination is 2000~10000Lux.
8. according to the method for claim 1, it is characterised in that: the gas that step (2) is passed through uses exhaust gas or flue gas, wherein
CO2Content is 10v%~45v%, and wherein v% is volume fraction；SO2Content is 200 × 10-6~600 × 10-6(v/v).
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| CN108384826B (en) * | 2018-05-25 | 2020-08-21 | 杭州富阳高博信息技术服务有限公司 | Process for preparing biodiesel by using single needle algae cells |
| CN117625397B (en) * | 2023-11-30 | 2024-06-14 | 山东省农业科学院 | Vermicelli wastewater treatment process based on microalgae |
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| CN103103130A (en) * | 2011-11-10 | 2013-05-15 | 中国石油化工股份有限公司 | Production method for lipid through mixed culture of microbes |
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| CN102250773A (en) * | 2011-05-31 | 2011-11-23 | 中国科学院青岛生物能源与过程研究所 | Scenedesmus as well as culturing method and application thereof |
| CN103103130A (en) * | 2011-11-10 | 2013-05-15 | 中国石油化工股份有限公司 | Production method for lipid through mixed culture of microbes |
| CN103184156A (en) * | 2011-12-29 | 2013-07-03 | 新奥科技发展有限公司 | Scenedesmus algal strain and its use |
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