CN102329826A - Method for producing grease rich in essential fatty acid linoleic acid and alpha-linolenic acid through culturing microalgae by use of CO2 - Google Patents
Method for producing grease rich in essential fatty acid linoleic acid and alpha-linolenic acid through culturing microalgae by use of CO2 Download PDFInfo
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Abstract
The invention provides a method for producing grease rich in essential fatty acid linoleic acid and alpha-linolenic acid through culturing microalgae by use of CO2. The method comprises the following steps: (1) screening and domesticating pollution-resisting algal species by utilizing a special screening culture system and a special culture medium under the high-efficiency high-CO2-concentration-resisting high-temperature-resisting conditions; (2) culturing domesticated microalgae by use of CO2; and (3) extracting the grease rich in linoleic acid and alpha-linolenic acid by taking the microalgae as a raw material. According to the invention, CO2 emitted by waste gas in a power plant is converted into a substance which can absorbed for growth of microalgae, so that algae cells with high biomass are obtained and high-value essential fatty acid linoleic acid and alpha-linolenic acid grease can be produced; and by using the method, the emission of CO2 is reduced, the greenhouse effect is alleviated, CO2 can be converted into high-value-added linoleic acid and alpha-linolenic acid grease, high yield of grease can be obtained, and the contents of linoleic acid and alpha-linolenic acid are maintained above 60%, thereby ensuring the quality of essential fatty acid grease and reducing the cost for producing the essential fatty acid grease.
Description
Technical field
The invention belongs to the renewable biological source field, relate to and utilize CO
2Culturing microalgae is produced essential fatty acids linoleic and the greasy method of alpha-linolenic acid of being rich in.
Background technology
The global warming that the greenhouse gases excessive emissions causes has become one of focus of national governments, academia and business circles concern.According to the data statistics of climate change administration of United Nations, greenhouse gases mainly are CO at present
2, account for 76.7%.In the world the concern of climate change has been caused the appearance of UNFCCC (UNFCCC), and the Kyoto Protocol of implementing in 2005 is born particularly CO of greenhouse gases to various countries
2The obligation that reduces discharging has been carried out clearly regulation.At present, country such as America and Europe has begun to impose CO
2Discharge fee, and through Clean Development Mechanism (Clean Development Mechanism, CDM) project to developing country provide with funds with technology carry out CO
2The emission rights transaction.Implement CO
2The reduction of discharging project can obtain the benefit of two aspects at least: the one, can obtain high economic benefit through the CDM project; The 2nd, reduce CO
2Quantity discharged can be improved the local climate condition, has considerable environmental benefit and social benefit.Therefore, on the basis of the energy demand of satisfying social development, effectively control CO
2Discharging has become one of various countries' major project to be solved.
Therefore photosynthetic microorganism can utilize inorganic carbon to be used for growth can absorb CO
2Be converted into living weight.CO
2The biological fixation method also is the most important and the most effective carbon fixation method on the present earth.Because little algae great majority are obligate ptotoautotroph, utilize inorganic carbon to be converted into biomass through photosynthesis, its intensity of photosynthesis substantially exceeds the metabolism total amount of the plant of equal quality.Therefore, through little algae photosynthesis fixation of C O
2Be considered to a kind of feasible and eco-friendly technology.Little in addition algae has growth cycle weak point, fast growth, CO
2Be considered to one of the most potential raw material of essential fatty acids linoleic and alpha-linolenic acid grease production Gu the carbon rate is high, indispensable fatty acid content is high, environmental resistance is strong, cultivate characteristics such as do not occupy cultivated land.
But linolic acid and alpha-linolenic acid are humans and animals healthy must generally the shortage, are badly in need of a kind of essential nutrients that replenishes.Wherein linolic acid has reduce fat, vessel softening, brings high blood pressure down, promotes microcirculatory effect; Can prevent or reduce the sickness rate of cardiovascular diseases; Particularly very favourable to the control of hypertension, hyperlipidemia, stenocardia, coronary heart disease, atherosclerosis, senile obesity etc.; Can play preventing the deposition of human serum SUV, the good reputation of " blood vessel street cleaner " arranged, health-care effect with the atherosclerosis of preventing and treating and cardiovascular disorder at vessel wall.Alpha-linolenic acid is the basic substance that constitutes cytolemma and enzyme, and HUMAN HEALTH is played a decisive role.In animal body through desaturation, increase two keys and prolong carbochain and be transformed into the essential composition of higher animal cell and have the timnodonic acid (EPA) and the 20 carbon acids (DHA) that prevent disease important physiological function such as cardiovascular and bring into play its important physical function.Alpha-linolenic acid has regulating blood fat, prevention myocardial infarction and cerebral infarction, reduction blood viscosity, increases the blood oxygen carrying content, to insulin resistant and mellitus, hypotensive, fat-reducing, suppress anaphylaxis, anti-inflammatory, vision protection, effect such as increase intelligence.U.S. FDA research proof: lack alpha-linolenic acid and will cause children's brain and retinal development slow, attention can not be concentrated, and nutrition is unbalanced; Can not effectively absorb, directly cause: MR, exercise not harmony; A little less than the eyesight, hyperkinetic syndrome, obesity; Apocleisis is grown slowly more than 30 kind of symptom such as hypoimmunity and disease.
Patent about essential fatty acids linoleic and alpha-linolenic acid production mainly is to concentrate on to utilize various land plants as raw material production linolic acid and alpha-linolenic acid at present.But because the basic material of these methods all is grain and oil crops, these terrestrial plant growth cycles are long, and growth is slow, and the soil is required than higher, and long-term benefit is not high.Can produce in addition with the people and strive a series of problems such as ground, crisis in food, be unfavorable for human Sustainable development.Utilize algae to produce gamma-linolenic acid to have only at present that a patent No. is 03113398.3, denomination of invention is the Chinese patent of " from the technology of algae extraction, preparation, purifying gamma-linolenic acid methyl esters ", it mainly focuses on and utilizes organic solvent and supercritical CO
2Method of extraction extracts gamma-linolenic acid from frustule.About utilizing CO
2Culturing microalgae is produced the patent or the blank out of indispensable fatty acid grease aspect.
Summary of the invention
Take all factors into consideration above each side, we have designed a kind of CO that utilizes in little algae absorbing waste gas
2Linolic acid and the greasy method of alpha-linolenic acid are rich in production as carbon source.Little algae carries out photosynthesis and absorbs CO
2After, the metabolism through self can be converted into lipid material (like lipid acid) in cell, then sophisticated microalgae cell is extracted the grease that is rich in linolic acid and alpha-linolenic acid through the organic extraction method.This not only greatly reduces carbon emission; Environment has been played good improvement effect; And can effectively reduce the cultivation cost of little algae; Also filled up simultaneously and utilized little algae to produce to be rich in essential fatty acids linoleic and the greasy technological vacancy of alpha-linolenic acid, for the full-scale development of biomass with fully utilize the new approach that provides.Therefore we can utilize and absorb CO
2Grow as carbon source, linolic acid and alpha-linolenic acid grease are rich in the frustule production of acquisition, make this research have the potentiality that reduce carbon emission and production high value and high added value useful matter simultaneously.
Therefore, the objective of the invention is to utilize CO
2The method of culturing microalgae production essential fatty acids linoleic and alpha-linolenic acid, specifically, detailed process of the present invention is included as efficient, anti-high CO
2The screening and the domestication of concentration, high temperature resistant and antipollution algae kind, and then the little algae after will taming is put into specific substratum and specific culture systems feeds different concns CO
2Cultivate, utilize the microalgae cell of cultivating acquisition to extract linolic acid and the alpha-linolenic acid grease that obtains to be rich in indispensable fatty acid, utilize response surface deathnium analysis optimization to obtain optimal culture condition at last through the organic extraction method.
The CO that utilizes of the present invention
2Culturing microalgae is produced and is rich in linolic acid and the greasy method of alpha-linolenic acid, may further comprise the steps:
(1) adopts particular screen culture apparatus, defined medium, efficient, anti-high CO
2Carry out the screening and the domestication of antipollution algae kind under concentration, the high temperature resistant condition;
(2) utilize CO
2Cultivate the little algae after taming;
(3) be that raw material extracts the grease that is rich in linolic acid and alpha-linolenic acid with the microalgae cell.
Particular screen culture systems in said (1) comprises bioreactor and gas distributing system; Said bioreactor is the ventilation bottle; The quantity of said bioreactor is 1-12, and said ventilation bottle connects said gas distributing system, and said gas distributing system provides different CO
2The device of concentration, it utilizes regulates air and CO
2Flow different CO recently are provided
2Concentration.
The volume of said ventilation bottle is 500mL-1L.
Said gas distributing system provides the different CO from 0.03%-50% respectively to said bioreactor simultaneously
2Concentration is screened little algae, the screening CO that provides
2Concentration is different because of the algae kind.
Specific substratum mainly is that carbon source Na is removed in the improvement back in said step (1)
2CO
3Improvement BG11 substratum.The prescription (every liter) of improvement BG11 substratum is as follows: 1mg Na
2EDTA, 36mg CaCl
22H
2O, 75mgMgSO
47H
2O, 40mg K
2HPO
43H
2O, 2.86mg H
3BO
3, 1.81mg MnCl
24H
2O, 0.222mgZnSO
47H
2O, 0.079mg CuSO
45H
2O, 0.05mg CoCl
26H
2O, 0.391mg NaMoO
42H
2O and 1500mgNaNO
3
Screening in said step (1) and acclimation method are for utilizing high concentration CO
2The screening and tame little algae, improve the high concentration CO of little algae
2Tolerance level and high temperature tolerance degree.CO
2Concentration range is 0.03%-50%, preferred 10%; The acclimation temperature scope is 20-40 ℃, preferred 30 ℃; The intensity of illumination scope is 4000lux-10000lux, preferred 8000lux; The ventilation flow rate scope is 0.1-1vvm, preferred 0.5vvm; The medium pH scope is 6.0-7.5, preferred 7.0.Concrete preferred parameter is different because of the algae kind.
Said antipollution algae kind is selected from a kind of in Chlorella pyrenoidesa (Chlorella pyrenoidosa), crescent moon algae (Selenastrum capricornutum), husky angle chlamydomonas (Chlamydomonas sajao), scenedesmus obliquus (Scenedesmus obliquus), chain silk algae (Hormidium sp.), aphanizomenon flos aquae (Aphanizo menon flos-aquae) and the collection ball algae (Palmellococcus chodat).
In said step (2), in particular B G11 substratum, feed the CO of different concns
2Carry out culturing microalgae.The inoculum density scope is 0.02-0.1g/L, preferred 0.05g/L; The intensity of illumination scope is 7000lux-9000lux, preferred 8000lux; CO
2Concentration range is 0.03%-50%, preferred 10%; The ventilation flow rate scope is 0.25-1vvm, preferred 0.5vvm; The culture temperature scope is 20-30 ℃, preferred 25 ℃; The medium pH scope is 6.0-7.5, preferred 7.0.Concrete preferred parameter is different because of the algae kind.
In said step (3), to adopt the organic extraction method be raw material with the microalgae cell through repeatedly extracting to such an extent that be rich in the grease of linolic acid and alpha-linolenic acid; Wherein organic extraction solvent is a chloroform: methyl alcohol (2: 1) or normal hexane; Consumption is every 1g frustule 20-80mL chloroform: the organic extraction agent of methyl alcohol (2: 1), preferred 60mL chloroform: the organic extraction agent of methyl alcohol (2: 1); The normal hexane consumption is every 1g frustule 20-100mL normal hexane, preferred 80mL normal hexane; Temperature of reaction 20-40 ℃, preferred 25 ℃; Reaction times 5-60 minute, preferred 30 minutes; Extraction times is 1-6 time, preferred 3 times.Concentrate (promptly revolve to steam and reclaim organic extraction agent) then and obtain grease; Said grease further utilizes GC-MS to analyze; Mainly contain 3 kinds of lipid acid: linolic acid, alpha-linolenic acid and palmitinic acid, wherein the content of two kinds of essential fatty acids linoleic and alpha-linolenic acid is respectively 13-20.7% and 40-48.4%.
Said method also further comprises utilizes response surface analysis to optimize culture condition.The main Minitab15 software that uses of response surface analysis experiment adopts the design that experimentizes of central composite design method, and independent variable is gas concentration lwevel and intensity of illumination, and response variable is living weight.The inoculum density scope is 0.02-0.1g/L, preferred 0.05g/L; The intensity of illumination scope is 2000lux-10000lux, preferred 7500lux; CO
2Concentration range is 0.03%-40%, preferred 10%; The ventilation flow rate scope is 0.25-1vvm, preferred 0.5vvm; The culture temperature scope is 20-30 ℃, preferred 25 ℃; The medium pH scope is 6.0-7.5, preferred 7.0.
Adopt the inventive method, not only can utilize the CO of power plant's exhaust gas emission
2Carbon source as little algae is cultivated has reached energy-saving and emission-reduction, alleviates the effect of Greenhouse effect, and can reduce the cost of production of biodiesel again, produces the essential fatty acids linoleic and the alpha-linolenic acid of high added value in addition.On the other hand; Method through organic extraction; Can obtain higher grease productive rate and keep linolic acid and the content of alpha-linolenic acid in the level more than 60%, not only this guarantees that the quality of indispensable fatty acid grease has also reduced the cost of producing the indispensable fatty acid grease.The waste gas CO of power plant
2Concentration be generally 10%, through the enforcement of this case, with this understanding, total fat content of little algae is 19.3%, therefore wherein the content of linolic acid and alpha-linolenic acid is respectively 19% and 41.54%, utilizes the waste gas CO of power plant
2The yield that culturing microalgae is produced linolic acid and alpha-linolenic acid can reach 3.66% and 8%.This is for the waste gas CO of power plant
2Comprehensive utilization and the production of high value essential fatty acids linoleic and alpha-linolenic acid all have positive effect.
Description of drawings
Fig. 1 representes to feed the CO of different concns
2Cultivate, the living weight of scenedesmus obliquus is with the variation of incubation time.
Wherein, X-coordinate is the time, and ordinate zou is the living weight of scenedesmus obliquus, and representes CO
2Concentration is 0.03%, ■ representes CO
2Concentration is 5%, zero expression CO
2Concentration is 10%, ● expression CO
2Concentration is 20%, △ representes CO
2Concentration is 30%, ▲ expression CO
2Concentration is 50%.Data are the MV of 3 parallel samplings.
Fig. 2 is illustrated in different concns CO
2Down, the solid carbon speed of scenedesmus obliquus is with the variation of incubation time.
Wherein, X-coordinate is the time, and ordinate zou is the solid carbon speed of scenedesmus obliquus, and representes CO
2Concentration is 0.03%, ■ representes CO
2Concentration is 5%, zero expression CO
2Concentration is 10%, ● expression CO
2Concentration is 20%, △ representes CO
2Concentration is 30%, ▲ expression CO
2Concentration is 50%.Data are the MV of 3 parallel samplings.
Fig. 3 representes different CO
2Concentration is to the influence of scenedesmus obliquus fat content.
Wherein, X-coordinate is CO
2Concentration, ordinate zou are the fat content of scenedesmus obliquus, and data are the MV of 3 parallel samplings.
Fig. 4 representes different CO
2Concentration is to the influence of the various fatty acid content of scenedesmus obliquus.
Wherein, X-coordinate is CO
2Concentration, ordinate zou are the various fatty acid content of scenedesmus obliquus, and data are the MV of 3 parallel samplings.
Fig. 5 representes the response surface figure of carbonic acid gas and light intensity reciprocal effect scenedesmus obliquus living weight.
Wherein, ordinate zou is the living weight of scenedesmus obliquus, and 2 coordinates of level are respectively CO
2Concentration and light intensity.
Fig. 6 representes the isogram of carbonic acid gas and light intensity reciprocal effect scenedesmus obliquus living weight.
Wherein, X-coordinate is CO
2Concentration, ordinate zou are light intensity.
Embodiment
Below describing is the mode description of a preferred embodiment thereof with instance, but not intention limits the invention.
Embodiment 1
(1) structure of particular screen culture systems: buy 250L illumination box, 6 air flowmeter, 6 CO from market
2Under meter, CO
2Gas measurement instrument, pneumatic pump, CO
2Steel cylinder, ventpipe, filtering membrane, special distribution support, special aerobic culture bottle, assembling constitute a cover can provide 6 kinds of different CO simultaneously
2The algae kind screening and culturing of concentration and domestication device.
(2) preparation of defined medium: owing to be CO mainly
2The crop carbon source is in order to tame little algae to CO
2Preference property, the defined medium of our configuration mainly is to remove carbon source Na
2CO
3The BG11 substratum.And by every liter of 1mg Na
2MGEDTA, 36mg CaCl
22H
2O, 75mg MgSO
47H
2O, 40mg K
2HPO
43H
2O, 2.86mg H
3BO
3, 1.81mg MnCl
24H
2O, 0.222mg ZnSO
47H
2O, 0.079mg CuSO
45H
2O, 0.05mg CoCl
26H
2O, 0.391mgNaMoO
42H
2O and 1500mg NaNO
3. consumption configuration defined medium, regulate pH value to 7.Substratum is sub-packed in the 1L aerobic culture bottle, every bottle of 800mL, 121 ℃ of high pressure steam sterilizations 20 minutes.
(3) efficient, anti-high CO
2The screening and the domestication of concentration, high temperature resistant and antipollution algae kind: respectively Chlorella pyrenoidesa (Chlorella pyrenoidosa), crescent moon algae (Selenastrum capricornutum), husky angle chlamydomonas (Chlamydomonas sajao), scenedesmus obliquus (Scenedesmus obliquus), chain silk algae (Hormidium sp.), aphanizomenon flos aquae (Aphanizo menon flos-aquae) and collection ball algae (Palmellococcus chodat) are inoculated in respectively in the sterilized defined medium; Place above-mentioned screening and domestication device to screen and cultivate then, feed 6 kinds of different CO
2Concentration: 0.03%, 5%, 10%, 20%, 30%, 50%, through screening and culturing, selecting scenedesmus obliquus (Scenedesmus obliquus) can be at high CO
2Growth fast under concentration and the higher culture temperature, living weight is maximum, and then scenedesmus obliquus is carried out the domestication of for some time.Through the 3-5 domestication in generation, scenedesmus obliquus has adapted to the high concentration CO that in no carbon source substratum, utilizes feeding
2The fast and stable growth.
(4) utilize CO
2Cultivate the little algae after the domestication: the scenedesmus obliquus after will taming (Scenedesmus obliquus) is cultivated, and chlorella is inoculated in the new special no carbon source BG11 substratum, keeps initial inoculum density about 0.5g/L; Because the waste gas CO of power plant
2Concentration range is 5%-15%, so we select 10%CO
2Culture temperature is 30 ± 1 ℃; Intensity of illumination is about 8000lux; Ventilation flow rate is about 0.25vvm; The medium pH value is about 7.0.Every at a distance from 1 day parallel sampling 3 times simultaneously with the living weight of spectrophotometric determination and dry weight method mensuration scenedesmus obliquus.Cultivate after 14 days when living weight and reach 1.84g/L, then with nutrient solution under 8000rpm centrifugal 5 minutes, collect the frustule lyophilize after, place crushing gently in the mortar, obtain exsiccant algae powder.
(5) be that raw material extracts the grease that is rich in linolic acid and alpha-linolenic acid with the microalgae cell: adopting organic extraction solvent is chloroform: methyl alcohol (2: 1) or normal hexane, and chloroform: it is every 1g frustule 60mL that the organic extraction agent of methyl alcohol (2: 1) gets consumption; The normal hexane consumption is every 1g frustule 80mL normal hexane; Temperature of reaction is 25 ℃; Reaction times is 30 minutes; Extracted several times is revolved then to steam and is reclaimed organic extraction agent, and the grease that obtains utilizes GC-MS to analyze again, mainly contains 3 kinds of lipid acid: linolic acid, alpha-linolenic acid and palmitinic acid.Wherein the content of two kinds of essential fatty acids linoleic and alpha-linolenic acid reaches 19% and 41.54% respectively.
Embodiment 2-6
To make an experiment, except the feeding CO of the scenedesmus obliquus after cultivating domestication with instance 1 similar methods
2Concentration is respectively 0.03%, 5%, 10%, 20%, 30%, 50%, other parameter constants.
Fig. 1 is illustrated in the CO of 5% and 10% concentration
2The living weight of following scenedesmus obliquus is maximum.Fig. 2 is illustrated in the CO of 5% and 10% concentration
2Following scenedesmus obliquus to CO
2Solid carbon speed maximum.Fig. 3 is illustrated in the CO of 50% concentration
2The fat content of following scenedesmus obliquus is the highest.Fig. 4 is illustrated in 0.03%, 5%, 10%, 20%, 30%, 50%CO
2Under the concentration, linoleic content is respectively 13%, 20.65%, 19%, 19.14%, 13.43%, 13.34%; And with the content of alpha-linolenic acid be respectively 43.35%, 40%, 41.54%, 40.67%, 48.44%, 48.23%, therefore at the CO of 5% above concentration
2The essential fatty acids linoleic of scenedesmus obliquus and the total content of alpha-linolenic acid are higher, all greater than 60%.
Embodiment 7
Adopt response surface analysis to optimize culture condition; Main Minitab 15 softwares that use of response surface analysis experiment; Design experimentizes to adopt central composite design method (being the analytical procedure in the Minitab software); Table 1 is meant the factor and the level code value of response surface experiment, and independent variable is gas concentration lwevel and intensity of illumination, and response variable is living weight.Central Composite test design and result are as shown in table 2.Inoculum density is 0.05g/L; Independent variable intensity of illumination scope is 2000lux-10000lux; Independent variable gas concentration lwevel scope is 0.03%-40%; Ventilation flow rate is 0.5vvm; Culture temperature is 30 ± 1 ℃; Medium pH is 7.0 ± 0.1.Fig. 5 and Fig. 6 are respectively response surface figure and the isograms that utilizes response surface carbonic acid gas and light intensity reciprocal effect scenedesmus obliquus living weight; The living weight that shows scenedesmus obliquus is interval at the waste gas gas concentration lwevel 6%-25% of power plant; Reach the highest in the interval 7.3-9.8klux of light intensity, wherein top condition is in 10% carbonic acid gas and 7500lux light intensity.
Table 1
Table 2
In sum, little algae absorbs CO
2Essential fatty acids linoleic and the greasy method of alpha-linolenic acid are rich in production as carbon source, not only can utilize the CO of power plant's exhaust gas emission
2Carbon source as little algae is cultivated has reached energy-saving and emission-reduction, alleviates the effect of Greenhouse effect, and can reduce the cost of production of biodiesel again, produces essential fatty acids linoleic and alpha-linolenic acid that content is higher than 60% high added value in addition.Also filled up and utilized CO
2Culturing microalgae is produced and is rich in essential fatty acids linoleic and the greasy technological vacancy of alpha-linolenic acid, for the full-scale development of biomass and comprehensive utilization provide a new approach this for the waste gas CO of power plant
2Comprehensive utilization and high value essential fatty acids linoleic and the greasy production of alpha-linolenic acid all have positive effect.
Patent related here and reference is all incorporated into here through reference with open, comprises all figure and table.
Claims (10)
1. utilize CO
2Culturing microalgae is produced and is rich in essential fatty acids linoleic and the greasy method of alpha-linolenic acid, it is characterized in that may further comprise the steps:
(1) adopts particular screen culture systems, defined medium, efficient, anti-high CO
2Carry out the screening and the domestication of antipollution algae kind under concentration, the high temperature resistant condition;
(2) utilize CO
2Cultivate the little algae after taming;
(3) be that raw material extracts the grease that is rich in linolic acid and alpha-linolenic acid with the microalgae cell.
2. the described CO that utilizes of claim 1
2Culturing microalgae is produced essential fatty acids linoleic and the greasy method of alpha-linolenic acid of being rich in; It is characterized in that the particular screen culture systems in said (1) comprises bioreactor and gas distributing system; Said bioreactor is the ventilation bottle; The quantity of said bioreactor is 1-12, and said ventilation bottle connects said gas distributing system, and said gas distributing system provides different CO
2The device of concentration, it utilizes regulates air and CO
2Flow different CO recently are provided
2Concentration.
3. the CO that utilizes according to claim 2
2Culturing microalgae is produced and is rich in essential fatty acids linoleic and the greasy method of alpha-linolenic acid, it is characterized in that said CO
2Range of concentrations is 0.03%-50%, when the quantity of said bioreactor greater than 1, said gas distributing system provides different CO respectively to said bioreactor simultaneously
2Concentration is screened little algae, the screening CO that provides
2Concentration is different because of the algae kind.
4. the CO that utilizes according to claim 1
2Culturing microalgae is produced and is rich in essential fatty acids linoleic and the greasy method of alpha-linolenic acid, it is characterized in that defined medium is for removing Na in said (1)
2CO
3The improvement BG11 substratum of carbon source.
5. the described CO that utilizes of claim 1
2Culturing microalgae is produced and is rich in essential fatty acids linoleic and the greasy method of alpha-linolenic acid, it is characterized in that said (1) middle and high concentration CO
2, high temperature screening and tame screening and the domestication CO of little algae
2Concentration range is 0.03%-50%; The acclimation temperature scope is 20-40 ℃; The intensity of illumination scope is 4000lux-10000lux; The ventilation flow rate scope is 0.1-1vvm; The medium pH scope is 6.0-7.5.
6. the described CO that utilizes of claim 5
2Culturing microalgae is produced and is rich in essential fatty acids linoleic and the greasy method of alpha-linolenic acid, it is characterized in that said CO
2Concentration range is 10%, and said acclimation temperature scope is 30 ℃, and said intensity of illumination scope is 8000lux, and said ventilation flow rate scope is 0.5vvm, and said medium pH scope is 7.0.
7. the described CO that utilizes of claim 1
2Culturing microalgae is produced and to be rich in essential fatty acids linoleic and the greasy method of alpha-linolenic acid, it is characterized in that said antipollution algae kind is selected from a kind of in Chlorella pyrenoidesa, crescent moon algae, husky angle chlamydomonas, scenedesmus obliquus, chain silk algae, aphanizomenon flos aquae and the collection ball algae.
8. the described CO that utilizes of claim 1
2Culturing microalgae is produced and is rich in essential fatty acids linoleic and the greasy method of alpha-linolenic acid, it is characterized in that cultured microalgae is a scenedesmus obliquus in said (2), and initial inoculum density is 0.5g/L, CO
2Concentration is 10%, and culture temperature is 30 ± 1 ℃, and intensity of illumination is 8000lux; Ventilation flow rate is 0.25vvm, and the medium pH value is 6.9-7.2, every at a distance from 1 day parallel sampling measure the living weight of scenedesmus obliquus for 3 times simultaneously; Cultivate after 14 days when living weight and reach 1.84g/L, then with nutrient solution under 8000rpm centrifugal 5 minutes, collect the frustule lyophilize after; Place crushing in the mortar, obtain exsiccant algae powder.
9. the described CO that utilizes of claim 1
2Culturing microalgae is produced essential fatty acids linoleic and the greasy method of alpha-linolenic acid of being rich in; It is characterized in that adopting in said (3) organic extraction solvent to extract; Said organic extraction solvent is a chloroform: methyl alcohol or normal hexane; Chloroform: the volume ratio of methyl alcohol is 2: 1, and this consumption that mixes organic extraction agent is the organic extraction agent of the dry algae powder 60mL of every 1g; The normal hexane consumption is every 1g frustule 80mL normal hexane; Extraction concentrates then and obtains grease, and said grease further utilizes GC-MS to analyze, and wherein the content of two kinds of essential fatty acids linoleic and alpha-linolenic acid is respectively 13-20.7% and 40-48.4%.
10. the described CO that utilizes of claim 1
2Culturing microalgae is produced essential fatty acids linoleic and the greasy method of alpha-linolenic acid of being rich in; It is characterized in that said method also further comprises utilizes response surface Central Composite analysis optimization culture condition; Said culture condition optimization utilizes gas concentration lwevel and intensity of illumination as independent variable; Living weight carries out the experimental design of response surface Central Composite as response variable, and wherein inoculum density is 0.05g/L; The intensity of illumination scope is 2000lux-10000lux; CO
2Concentration range is 0.03%-40%; Ventilation flow rate is 0.5vvm; Culture temperature is 30 ℃; Medium pH is 7.0.
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| CN201110299976A Pending CN102329826A (en) | 2011-09-28 | 2011-09-28 | Method for producing grease rich in essential fatty acid linoleic acid and alpha-linolenic acid through culturing microalgae by use of CO2 |
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| CN102703325A (en) * | 2012-06-28 | 2012-10-03 | 广西科学院 | Method for producing chlamydomonas at low cost |
| CN105713951A (en) * | 2014-12-05 | 2016-06-29 | 中国石油化工股份有限公司 | Method for preparing microalgae oil |
| CN108587913A (en) * | 2017-06-08 | 2018-09-28 | 中国科学院水生生物研究所 | A kind of scenedesmus, its cultural method and its application with high ALA contents |
| KR20190136190A (en) | 2018-05-30 | 2019-12-10 | 한국생명공학연구원 | Novel microalgae of Lobosphaera incisa K-1 and its use |
| CN112973434A (en) * | 2021-03-08 | 2021-06-18 | 青岛大学 | Phase change solvent reinforced microalgae fixed coal-fired flue gas CO2And resource conversion method |
| CN116076405A (en) * | 2022-12-30 | 2023-05-09 | 中国科学院青岛生物能源与过程研究所 | Method for cultivating fish, shrimp and shellfish and artemia by utilizing nervonic acid microalgae |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102703325A (en) * | 2012-06-28 | 2012-10-03 | 广西科学院 | Method for producing chlamydomonas at low cost |
| CN105713951A (en) * | 2014-12-05 | 2016-06-29 | 中国石油化工股份有限公司 | Method for preparing microalgae oil |
| CN105713951B (en) * | 2014-12-05 | 2020-04-14 | 中国石油化工股份有限公司 | Method for preparing microalgae grease |
| CN108587913A (en) * | 2017-06-08 | 2018-09-28 | 中国科学院水生生物研究所 | A kind of scenedesmus, its cultural method and its application with high ALA contents |
| CN108587913B (en) * | 2017-06-08 | 2021-09-03 | 中国科学院水生生物研究所 | Scenedesmus with high alpha-linolenic acid content, and culture method and application thereof |
| KR20190136190A (en) | 2018-05-30 | 2019-12-10 | 한국생명공학연구원 | Novel microalgae of Lobosphaera incisa K-1 and its use |
| CN112973434A (en) * | 2021-03-08 | 2021-06-18 | 青岛大学 | Phase change solvent reinforced microalgae fixed coal-fired flue gas CO2And resource conversion method |
| CN112973434B (en) * | 2021-03-08 | 2023-08-11 | 青岛大学 | Phase-change solvent-reinforced microalgae-immobilized coal-fired flue gas CO 2 And a method of converting into resources |
| CN116076405A (en) * | 2022-12-30 | 2023-05-09 | 中国科学院青岛生物能源与过程研究所 | Method for cultivating fish, shrimp and shellfish and artemia by utilizing nervonic acid microalgae |
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