CN107460217A - A kind of method that mixed culture prepares microalgae grease - Google Patents

A kind of method that mixed culture prepares microalgae grease Download PDF

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CN107460217A
CN107460217A CN201610386731.3A CN201610386731A CN107460217A CN 107460217 A CN107460217 A CN 107460217A CN 201610386731 A CN201610386731 A CN 201610386731A CN 107460217 A CN107460217 A CN 107460217A
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algae
fsh
culture
microalgae
micro
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CN107460217B (en
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师文静
孙启梅
廖莎
王鹏翔
李晓姝
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China Petroleum and Chemical Corp
Sinopec Fushun Research Institute of Petroleum and Petrochemicals
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Sinopec Fushun Research Institute of Petroleum and Petrochemicals
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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
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6436—Fatty acid esters
    • C12P7/649—Biodiesel, i.e. fatty acid alkyl esters
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    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
    • B01D53/34—Chemical or biological purification of waste gases
    • B01D53/46—Removing components of defined structure
    • B01D53/54—Nitrogen compounds
    • B01D53/56—Nitrogen oxides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
    • B01D53/34—Chemical or biological purification of waste gases
    • B01D53/46—Removing components of defined structure
    • B01D53/62—Carbon oxides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
    • B01D53/34—Chemical or biological purification of waste gases
    • B01D53/74—General processes for purification of waste gases; Apparatus or devices specially adapted therefor
    • B01D53/84—Biological processes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D2251/00—Reactants
    • B01D2251/95—Specific microorganisms
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D2256/00—Main component in the product gas stream after treatment
    • B01D2256/22—Carbon dioxide
    • 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
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
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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
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    • 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
    • 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 method that mixed culture prepares microalgae grease, including following content：Micro-algae culture medium and mixing microalgae seed liquor are added in bioreactor, it is 8~12, preferably 9~11 to maintain cultivating system PH, and is passed through the CO that volume content is 5v%~45v%2, preferably 10v%~30v%；Wherein mixing microalgae includes kelvin plan chlorella（Parachlorella kessleri）FSH Y3 and algae fibre（Ankistrodesmus sp.）SS B7, it is preserved in " China Committee for Culture Collection of Microorganisms's common micro-organisms center " respectively on May 26th, 2014 and on April 15th, 2013, deposit number is respectively CGMCC No.9238 and CGMCC No.7478.The inventive method improves microdisk electrode system to high concentration CO2Tolerance and dissolubility, improve carbon sequestration efficiency, the harvest yield of microalgae grease significantly improves, be advantageous to carry out biodiesel production.

Description

A kind of method that mixed culture prepares microalgae grease
Technical field
The invention belongs to biotechnology and field of biological energy source, and in particular to a kind of mixed culture prepares the side of microalgae grease Method.
Background technology
Due to the increase for reducing increasingly and greenhouse effects being caused using fossil energy of fossil energy, increasing scientific research Worker focuses on sight in the development and utilization of regenerative resource.Biomass energy is as most important renewable energy on the earth Source, it includes forestry biomass, crops, water plant, agricultural wastes etc..In many biomass energies, microalgae is Important renewable resource.They have widely distributed, biomass is big, photosynthetic efficiency is high, strong environmental adaptability, growth The features such as cycle is short, biomass yield is high.Primary or secondary metabolite containing uniqueness in its cell, complex chemical composition.It is micro- The solar conversion efficiency of algae can reach 3.5%, be the potential resource for producing medicine, fine chemical product and New-type fuel, from microalgae In obtained aliphatic acid can change into Fatty acid methyl ester, i.e. biodiesel.
With the development of World Economics, substantial amounts of fossil energy using and consuming, and causes shortage and the environment of the energy Worsening, particularly 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 more than 10 times of terrestrial plant under certain condition, can not only reduce CO2Discharge, simultaneously Also reduce toxigenic capacity；Except CO2Outside, the composition such as some SOx, NOx in waste gas is cleaned place also with the metabolism of microalgae Reason, noxious gas emission is effectively reduced, therefore the biodiesel produced by the use of microalgae grease as raw material is most possible at present Meet the regenerative resource of fuel needed for world's transport.
At present for the more of the oil-producing microalgaes such as chlorella, grid algae research.CN20110144545.6 discloses one plant of grid algae Algae strain, the growth of the algae strain can utilize synthetic medium or appropriately processed waste water to grow, be characterized in that lipid-producing is higher than Most of at present to divide algae strain, the 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 one plant of rich oil marine microalgae it is micro- intend ball algae (Nannochloropsis gaditana ) algae strain and its application, the algae strain can in the environment of pH=4.5 normal growth, its oil Fat content is up to 35%.CN20111019480.X discloses one plant of microalgae algae strain（Mychonases sp .）And its for producing The application of biodiesel, the polyunsaturated fatty acid of high added value, including leukotrienes C18 can be produced using the algae strain:3 and god Through sour C24:1, it obtains the byproduct of high added value while biodiesel is obtained.These patents are all not directed to algae kind pair The tolerance of carbon dioxide.CN102703326A discloses a kind of high CO2The microalgae and its selection of tolerance and fixed rate, But the algae strain that the patent is provided is not directed to the fat content of the algae strain.
Or above-mentioned patent can not efficiently utilize CO2Grease is produced, otherwise fat content is not high enough in the biomass obtained. Particularly in actual applications, as CO in environment2When volume fraction is more than 5v%, the growth of most of microalgae will be suppressed, Gu Carbon efficiencies are low；General microalgae suitable growth in neutral conditions simultaneously, is unfavorable for micro- under conditions of slant acidity or meta-alkalescence The growth of algae, and microalgae utilizes CO2HCO usually to be dissolved in water3 -Existing for ionic species, carbon dioxide is in neutral ring Solubility is low under border, is unfavorable for algae and absorbs.If moreover, the SOx containing high concentration in the fossil fuel waste gas being passed through, The gases such as NOx can also suppress micro algae growth and reduce carbon sequestration efficiency.
The content of the invention
In view of the shortcomings of the prior art, the present invention provides a kind of method that mixed culture prepares microalgae grease.Present invention side Method improves microdisk electrode system to high concentration CO2Tolerance and dissolubility, improve carbon sequestration efficiency, the harvest of microalgae grease Amount significantly improves, and is advantageous to carry out the production of biodiesel.
The method that mixed culture of the present invention prepares microalgae grease, including following content：Added in bioreactor micro- Algae culture medium and mixing microalgae seed liquor, it is 8~12, preferably 9~11 to maintain cultivating system PH, and is passed through volume content and is 5v%~45v% CO2, preferably 10v%~30v%；Wherein mixing microalgae includes kelvin plan chlorella（Parachlorella kessleri）FSH-Y3 and algae fibre（Ankistrodesmus sp.）SS-B7, respectively on May 26th, 2014 and 2013 4 The moon is preserved in " China Committee for Culture Collection of Microorganisms's common micro-organisms center " on the 15th, and deposit number is respectively CGMCC No. 9238 and CGMCC No. 7478, preservation address are that the Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Chinese Academy of Sciences is micro- Biological study institute.
In the present invention, described kelvin intend chlorella FSH-Y3 algaes strains under the microscope frustule be it is spherical, oval, it is interior There is the chromatoplast of a Zhousheng, cup-shaped or sheet；Vegetative propagation, each cell can produce 2,4,8 or 16 autospores, into Mother cell ruptures when ripe, spore effusion, is new individual after growing up.The algae strain can preferably absorb two under high pH value Carbonoxide, fast-growth breeding.Described algae fibre SS-B7 algae strains are a kind of green algate of fresh water, and frustule is green under the microscope Color, crescent or arch, grow thickly, bend, tapering thin from mediad both ends, distal tip is long about 5-6 μm, and center is wide about 2 μm.Should Algae strain is resistant to the CO of high concentration2And NOx, it can utilize and contain CO2Illumination autophyting growth is carried out with NOx waste gas or flue gas to obtain Take rich grease-contained biomass, carbon sequestration efficiency high, tolerance is strong.
In the present invention, micro-algae culture medium cultivates the liquid of microalgae using BG11, SE, BBM well known to those skilled in the art etc. Body culture medium.
In the present invention, it is as follows that kelvin intends preparing for chlorella FSH-Y3 seed liquors：The PH of culture medium is adjusted to 10~12, It it is 20~30 DEG C, periodicity of illumination 24h in temperature, brightness time ratio is 14:10, intensity of illumination is 2000~20000Lux, is shaken Culture is swung to exponential phase.The kelvin that is added in bioreactor intends chlorella FSH-Y3 seed liquors and micro-algae culture medium Volume ratio 1:25~1:5.
In the present invention, preparing for algae fibre SS-B7 seed liquors is as follows：The PH of culture medium is adjusted to 7~9, is in temperature 20~30 DEG C, periodicity of illumination 24h, brightness time ratio is 14:10, intensity of illumination is 2000~10000Lux, and shaken cultivation is extremely Exponential phase.The algae fibre SS-B7 seed liquors and the volume ratio of micro-algae culture medium added in bioreactor is 1:25~ 1:5。
In the present invention, the scenedesmus obliquus described in a certain amount of CN201310537896.2 is preferably added in co-culture system （Scenedesmus obliqnus）FSH-Y2, the algae strain were preserved in " Chinese microorganism strain preservation pipe on 11st in September in 2012 Reason committee common micro-organisms center ", deposit number are CGMCC No. 6551, and preservation address is Chaoyang District, Beijing City North Star west No. 3 Institute of Microorganism, Academia Sinica of institute of road 1.The preparation method of scenedesmus obliquus FSH-Y2 seed liquors intends bead with kelvin Algae FSH-Y3, addition are scenedesmus obliquus FSH-Y2 seed liquors and the volume ratio 1 of micro-algae culture medium:40~1:10.
In the present invention, the temperature of mixed culture is 20~30 DEG C, periodicity of illumination 24h, and brightness time ratio is 14:10, light It is 2000~10000Lux according to intensity.It is passed through gas and preferably uses and contains CO2Waste gas or flue gas, wherein CO2Content be 10v%~ 45v%, NOx content are not higher than 500 × 10-6（v/v）.Mixed culture to growth stationary phase terminates, by the mode such as centrifuging, settling Harvesting microalgae cell, determines dry cell weight and fat content, dry cell weight can reach more than 12g/L, and fat content can reach carefully More than the 45% of born of the same parents' dry weight.
Compared with prior art, the present invention can bring following beneficial effect：
1st, kelvin is intended chlorella FSH-Y3 and algae fibre SS-B7 mixed culture by the present invention, improves microdisk electrode system to height Concentration C O2Tolerance and carbon sequestration efficiency, the harvest yield of microalgae grease significantly improves.
2nd, the present invention carries out the culture of microalgae under high PH environment, by increasing capacitance it is possible to increase CO in cultivating system2Solubility, help In microalgae to CO2Absorb, improve carbon sequestration efficiency.Meanwhile in high pH value culture microalgae, it can effectively suppress microalgae The growth of miscellaneous bacteria in growth course, improve microalgae oil yield.
3rd, co-culture system of the invention is resistant to the CO of high concentration2And NOx, the CO in waste gas can be utilized2Carry out Autophyting growth, fixed CO2, alleviate greenhouse effects and exhaust pollution problems that current industrial society brings.
Embodiment
The present invention is described in further detail below by embodiment.In the present invention, wt% is mass fraction, and v% is body Fraction.
The preparation of the microalgae seed liquor of embodiment 1
Microdisk electrode uses BG11 culture mediums, and culture medium prescription is as shown in Table 1 and Table 2.
The BG11 culture mediums of table 1
* in the table 1 of table 2 A5+Co solution composition
It is first according to Tables 1 and 2 and prepares BG11 fluid nutrient mediums, culture kelvin is intended into chlorella FSH-Y3, scenedesmus obliquus FSH- The PH of Y2 culture medium is adjusted to 10, and the PH for the culture medium for cultivating algae fibre SS-B7 is adjusted into 8.0, then intended kelvin small Ball algae FSH-Y3, scenedesmus obliquus FSH-Y2, algae fibre SS-B7 are inoculated in above-mentioned culture medium respectively.In constant temperature illumination shaking table Culture, cultivation temperature are 25 DEG C, periodicity of illumination 24h, and brightness time ratio is 14:10, intensity of illumination 5000Lux, 120rpm Shaken cultivation obtains kelvin and intends chlorella FSH-Y3 seed liquors, scenedesmus obliquus FSH-Y2 seed liquors, algae fibre to exponential phase SS-B7 seed liquors, above-mentioned seed liquor is saved backup under 15 DEG C of dim lights.
The preparation of the microalgae grease of embodiment 2
Microalgae seed liquor prepared by embodiment 1 is seeded in the production of BG11 culture mediums progress microalgae grease respectively.In 20L gas lifts Carried out in formula bioreactor, kelvin intends chlorella FSH-Y3 seed liquors, algae fibre SS-B7 seed liquors according to volume ratio 1:1 Ratio inoculation, be inoculated into according to 20% total inoculum concentration in BG11 culture mediums, pH value control between 8~11, be passed through nitrogen and The mixed gas of carbon dioxide, wherein carbon dioxide content are 10v%~45v%, intensity of illumination 5000Lux, and cultivation temperature is 28 DEG C, periodicity of illumination 24h, brightness time ratio is 14:10.Culture is in stationary phase after 7 days, terminate culture, algae is collected by centrifugation Liquid, algae dried bean noodles weight is measured after vacuum freeze drying to constant weight under the conditions of -60 DEG C, calculate yield of biomass, and use n-hexane: Ethyl acetate method measures total lipid content, as a result as shown in table 3.
The mixed algae of table 3 is compared with the culture effect of single algae
From table 3, relative to single algae kind, mixed algae kind is in high PH and high CO2Under concentration, obtain higher biomass and Fat content.
The preparation of the microalgae grease of embodiment 3
Scenedesmus obliquus FSH-Y2 seed liquors are added in cultivating system, wherein kelvin intends chlorella FSH-Y3 seed liquors, tiltedly gives birth to grid Algae FSH-Y2 seed liquors, algae fibre SS-B7 seed liquors three is inoculated into BG11 culture mediums according to total inoculum concentration 20%.It is other With embodiment 2, effect is as shown in table 4 for process and condition.
The mixed algae culture effect of table 4 compares
From table 4, scenedesmus obliquus FSH-Y2 seed liquors are added in mixed system, the effect of cultivating system, which has, further to be carried It is high.
Embodiment 4 prepares microalgae grease using flue gas
Preparation condition is same as Example 2, and difference is to be passed through containing NO, NO2And CO2Flue gas, CO in flue gas2Content For 10v%~40v%, NO and NO2Content is 100 × 10-6~500 × 10-6（v/v）, the PH of maintenance reaction system is 10.After 10 days Terminate culture, frustule is collected by centrifugation, algae dried bean noodles weight is measured after vacuum freeze drying to constant weight under the conditions of -60 DEG C, calculate life Substance production, and use n-hexane:Ethyl acetate method measures total lipid content, as a result as shown in table 5.
The mixed algae of table 5 is compared with the culture effect of single algae
From table 5, relative to single algae kind, mixed algae kind not only can be with the CO of enduring high-concentration2, and can be resistant to certain The NOx of concentration, obtain higher biomass and fat content.As can be seen here, mixed algae and flue gas can be utilized to prepare microalgae Grease, that is, the production of grease is realized, while waste gas can be purified.

Claims (10)

1. a kind of method that mixed culture prepares microalgae grease, it is characterised in that including following content：In bioreactor Add micro-algae culture medium and mixing microalgae seed liquor, it is 8~12 to maintain cultivating system PH, and be passed through volume content for 5v%~ 45v% CO2, wherein mixing microalgae, which includes kelvin, intends chlorella（Parachlorella kessleri）FSH-Y3 and algae fibre （Ankistrodesmus sp.）SS-B7, " China Microbiological is preserved in respectively on May 26th, 2014 and on April 15th, 2013 Culture presevation administration committee common micro-organisms center ", deposit number are respectively CGMCC No. 9238 and CGMCC No. 7478。
2. according to the method for claim 1, it is characterised in that：Described kelvin intends chlorella FSH-Y3 algae strains in microscope Lower frustule is spherical, oval, inside there is the chromatoplast of a Zhousheng, cup-shaped or sheet；Vegetative propagation, each cell can produce Raw 2,4,8 or 16 autospores, mother cell ruptures when ripe, spore effusion, is new individual after growing up；Described algae fibre SS-B7 algae strains are a kind of green algate of fresh water, and frustule is green under the microscope, crescent or arch, grows thickly, bends, from mediad Both ends are tapering thin, and distal tip is long about 5-6 μm, and center is wide about 2 μm.
3. according to the method for claim 1, it is characterised in that：Micro-algae culture medium is using BG11, SE or BBM culture microalgae Fluid nutrient medium.
4. according to the method for claim 1, it is characterised in that：It is as follows that kelvin intends preparing for chlorella FSH-Y3 seed liquors： The PH of culture medium is adjusted to 10~12, temperature is 20~30 DEG C, periodicity of illumination 24h, and brightness time ratio is 14:10, illumination Intensity is 2000~20000Lux, shaken cultivation to exponential phase.
5. the method according to claim 1 or 4, it is characterised in that：The kelvin added in bioreactor intends chlorella FSH-Y3 seed liquors and the volume ratio of micro-algae culture medium 1:25~1:5.
6. according to the method for claim 1, it is characterised in that：Preparing for algae fibre SS-B7 seed liquors is as follows：By culture medium PH be adjusted to 7~9, temperature be 20~30 DEG C, periodicity of illumination 24h, brightness time ratio be 14:10, intensity of illumination is 2000~10000Lux, shaken cultivation to exponential phase.
7. the method according to claim 1 or 6, it is characterised in that：The algae fibre SS-B7 kinds added in bioreactor The volume ratio of sub- liquid and micro-algae culture medium is 1:25~1:5.
8. according to the method for claim 1, it is characterised in that：CN201310537896.2 is added in co-culture system Described scenedesmus obliquus（Scenedesmus obliqnus）FSH-Y2；The preparation method of scenedesmus obliquus FSH-Y2 seed liquors is the same as triumphant Family name intends chlorella FSH-Y3, and addition is scenedesmus obliquus FSH-Y2 seed liquors and the volume ratio 1 of micro-algae culture medium:40~1:10.
9. according to the method for claim 1, it is characterised in that：It is 20~30 DEG C to be mixed temperature, and periodicity of illumination is 24h, brightness time ratio are 14:10, intensity of illumination is 2000~10000Lux, after cultivating system enters stationary phase, terminates culture.
10. the method according to claim 1 or 9, it is characterised in that：Mixed culture, which is passed through gas and used, contains CO2Waste gas or Flue gas, wherein CO2Content is 10v%~45v%, and NOx content is not higher than 500 × 10-6（v/v）.
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CN111100885A (en) * 2018-10-26 2020-05-05 中国石油化工股份有限公司 Method for improving oil production of microalgae
CN111100883A (en) * 2018-10-26 2020-05-05 中国石油化工股份有限公司 Method for producing microalgae grease by using flue gas
CN111100883B (en) * 2018-10-26 2022-08-12 中国石油化工股份有限公司 Method for producing microalgae grease by using flue gas

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