CN104293473A - Method for extracting oil from oil production microbes - Google Patents
Method for extracting oil from oil production microbes Download PDFInfo
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- CN104293473A CN104293473A CN201310306509.4A CN201310306509A CN104293473A CN 104293473 A CN104293473 A CN 104293473A CN 201310306509 A CN201310306509 A CN 201310306509A CN 104293473 A CN104293473 A CN 104293473A
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- oleaginous microorganism
- grease
- slurry
- oil
- microorganism
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/10—Production of fats or fatty oils from raw materials by extracting
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/12—Production of fats or fatty oils from raw materials by melting out
- C11B1/14—Production of fats or fatty oils from raw materials by melting out with hot water or aqueous solutions
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Fats And Perfumes (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a method for extracting oil from oil production microbes. The method is characterized in that the method comprises the following steps: 1, adjusting the pH value of a water slurry containing the oil production microbes to 1-5; 2, stirring under hydrothermal conditions to dissolve out oil in the cells of the oil production microbes in order to obtain an oil production microbes dissolved slurry containing oil; and 3, separating the oil from the oil production microbes dissolved slurry. The method for extracting oil from oil production microbes allows oil in the cells of the oil production microbes to be dissolved out when the pH value is 1-5, and the water phase and the oil phase of the obtained oil production microbes dissolved slurry are easy to separate, so the oil extraction rate is high.
Description
Technical field
The present invention relates to oleaginous microorganism field of engineering technology, particularly, relate to a kind of method extracting grease from oleaginous microorganism.
Background technology
Because microorganism cells propagation is fast; with short production cycle; microorganism growth desired raw material wide material sources; low price, and microorganism growth does not limit by season, climate change, energy scale operation continuously; production cost is low; and oil length is high in oleaginous microorganism, therefore, utilize oleaginous microorganism to produce grease and become a kind of important means alleviating global energy crisis.
Micro-algae is a kind of aquatic phototroph, and it utilizes luminous energy, with CO
2and H
2o carries out the growth of cell as substrate, and in cell, synthesize a large amount of materials such as triglyceride level (TAGs), polysaccharide, protein.Wherein, triglyceride level (TAGs) comprises unsaturated glyceride (DHA, EPA) not only can as important dietary supplements, and can be used as the important glyceride stock source of biological liquid fuel.Particularly along with world energy sources is as the day by day shortage of oil, with animal-plant oil (comprising microalgae grease) for raw material production biofuel is subject to extensive concern, and competitively carry out the research of microalgae biodiesel.
The extraction of microalgae grease is the gordian technique of biological diesel oil refining always.Micro-algae of most of kind has cellulosic cell walls, the microalgae cell wall of some kind is difficult to other compositions of digest and decompose in addition in a large number, cell wall thickness, strong but pliable in texture, be unfavorable for very much the extraction of grease and nutritive substance in cell, need to carry out broken wall treatment to micro-algae.
The extraction of current microalgae grease adopts hydrothermal method usually.Hydrothermal method normally carries out cell wall breaking, grease stripping by High Temperature High Pressure in the basic conditions, general temperature is 100-300 DEG C, the molten slurry of the micro-algae of gained carries out oily water separation by centrifugal, as CN100460513C, but due to the milk sap that the molten slurry of micro-algae obtained is containing algae-residue, oil phase and aqueous phase mix, and are difficult to be separated, and grease extraction efficiency is lower.
Therefore, the efficient and technology extracting grease from oleaginous microorganism of economy becomes the emphasis of research.
Summary of the invention
The object of the invention is to overcome in prior art from oleaginous microorganism, extract the low defect of grease extraction efficiency, a kind of method extracting grease efficiently from oleaginous microorganism is provided.
The present inventor surprisingly finds under study for action, and be the grease stripping made under the condition of 1-5 in oleaginous microorganism cell in pH value, aqueous phase and the oil phase of the molten slurry of the oleaginous microorganism obtained are easily separated, and grease extraction efficiency is high.
Therefore, to achieve these goals, the invention provides a kind of method extracting grease from oleaginous microorganism, it is characterized in that, described method comprises the steps:
(1) aqueous slurry to the pH value containing oleaginous microorganism is regulated to be 1-5;
(2) stir under hydrothermal conditions, make the grease stripping in oleaginous microorganism cell, obtain the molten slurry of oleaginous microorganism containing grease;
(3) from the molten slurry of oleaginous microorganism, grease is isolated.
Preferably, in step (1), aqueous slurry to the pH value containing oleaginous microorganism is regulated to be 2-4.
Preferably, with the dry weight basis of oleaginous microorganism, described is 2-40 % by weight containing the content of oleaginous microorganism in the aqueous slurry of oleaginous microorganism.
Preferably, in step (2), hydrothermal condition comprises: temperature is 120-400 DEG C, is preferably 160-260 DEG C.
Preferably, in step (2), the condition of stirring comprises: the material be stirred relative to every cubic metre, and power of agitator is 0.15-1.5kW, is preferably 0.2-1kW; Churning time is 2-250 minute, is preferably 30-120 minute.
Preferably, in step (3), the method isolating grease from the molten slurry of oleaginous microorganism comprises centrifugation, and the condition of described centrifugation comprises: rotating speed 3000-8000rpm, time 2-10 minute.
Preferably, in step (3), the method isolating grease from the molten slurry of oleaginous microorganism comprises and being contacted with extraction agent by molten for described oleaginous microorganism slurry, extracts grease.
Preferably, with the dry weight basis of oleaginous microorganism, relative to 1g oleaginous microorganism, the consumption of described extraction agent is 5-20mL.
Preferably, described extraction agent is selected from one or more in hexane, methylene dichloride, trichloromethane, tetrachloromethane, toluene and tetrahydrofuran (THF).
Preferably, described oleaginous microorganism is micro-algae.Described micro-algae is preferably selected from least one in chlorella, grid algae, spirulina, chrysophyceae and Phaeodactylum tricornutum.
The method extracting grease from oleaginous microorganism provided by the invention, be the grease stripping made under the condition of 1-5 in oleaginous microorganism cell in pH value, aqueous phase and the oil phase of the molten slurry of the oleaginous microorganism obtained are easily separated, and grease extraction efficiency is high.
Other features and advantages of the present invention are described in detail in embodiment part subsequently.
Embodiment
Below the specific embodiment of the present invention is described in detail.Should be understood that, embodiment described herein, only for instruction and explanation of the present invention, is not limited to the present invention.
The invention provides a kind of method extracting grease from oleaginous microorganism, the method comprises the steps:
(1) aqueous slurry to the pH value containing oleaginous microorganism is regulated to be 1-5;
(2) stir under hydrothermal conditions, make the grease stripping in oleaginous microorganism cell, obtain the molten slurry of oleaginous microorganism containing grease;
(3) from the molten slurry of oleaginous microorganism, grease is isolated.
According to the present invention, although adopt aforesaid method can realize object of the present invention, even if the aqueous phase of the molten slurry of oleaginous microorganism and oil phase easily separated, grease extraction efficiency is high.But under preferable case, in step (1), regulate the aqueous slurry containing oleaginous microorganism to be 2-4 to pH value, can further improve grease extraction efficiency.
In the present invention, the aqueous slurry containing oleaginous microorganism refers to the liquid containing oleaginous microorganism collected oleaginous microorganism and obtain from oleaginous microorganism nutrient solution.With the dry weight basis of oleaginous microorganism, be preferably 2-40 % by weight containing the content of oleaginous microorganism in the aqueous slurry of oleaginous microorganism, be more preferably 7-15 % by weight.
In the present invention, for pH value condition, can acid adding regulate, for acid without particular requirement, the acid that this area is conventional can be adopted, such as, acid can be selected from sulfuric acid, hydrochloric acid, formic acid, acetic acid, citric acid, lactic acid, toxilic acid, tartrate and oxalic acid one or more.
In the present invention, pH value is recorded by pH meter.
In the present invention, hydrothermal condition refers to and keeps liquid condition at a certain temperature.In step of the present invention (2), hydrothermal condition preferably includes: temperature is 120-400 DEG C; More preferably comprise: temperature is 160-260 DEG C.
It will be understood by those skilled in the art that in step (2), as long as the pressure of hydrothermal condition makes water can keep liquid at the temperature disclosed above.
In step of the present invention (2), for the condition stirred, as long as make the grease stripping in oleaginous microorganism cell.Such as, the condition of stirring can comprise: the material be stirred relative to every cubic metre, and power of agitator is 0.15-1.5kW; Churning time is 2-250 minute; Preferably, the condition of stirring can comprise: the material be stirred relative to every cubic metre, and power of agitator is 0.2-1kW; Churning time is 30-120 minute.Under above-mentioned agitation condition, the grease stripping in oleaginous microorganism cell can be made.
In the present invention, namely the molten slurry of oleaginous microorganism refers to oleaginous microorganism cell wall damage in the aqueous slurry containing oleaginous microorganism, and born of the same parents' inner structure is disintegrated, the mixture that the stripping of oleaginous microorganism grease obtains.
In step of the present invention (3), grease is isolated from the molten slurry of oleaginous microorganism, hydrothermal method can be adopted to extract method conventional in grease, such as centrifugation, but the aqueous phase of the molten slurry of oleaginous microorganism obtained due to the inventive method and oil phase easily separated, therefore, relative to traditional hydrothermal method, can reduce the energy consumption of centrifugation, the condition optimization of centrifugation comprises: rotating speed 3000-8000rpm, time 2-10 minute; More preferably comprise: rotating speed 4000-6000rpm, time 4-6 minute.
Aqueous phase and the oil phase of the molten slurry of oleaginous microorganism obtained due to the inventive method are easily separated, therefore, except the centrifugal separation method used in conventional hydrothermal method, molten for oleaginous microorganism slurry can also contact with extraction agent by the method isolating grease from the molten slurry of oleaginous microorganism, extracts grease.
In the present invention, contacted by molten for oleaginous microorganism slurry with extraction agent, namely the method extracting grease is contacted with extraction agent by molten for oleaginous microorganism slurry, abundant mixing, then stratification, collect extract layer, utilize extraction agent boiling point lower than the character of grease, evaporate to dryness extraction agent, thus obtain grease.
In the present invention, with the dry weight basis of oleaginous microorganism, relative to 1g oleaginous microorganism, the consumption of extraction agent is preferably 5-20mL; Be more preferably 5-10mL.
In the present invention, for extraction agent without particular requirement, the extraction agent that oil and grease extracting is conventional can be adopted, such as, extraction agent can be selected from hexane, methylene dichloride, trichloromethane, tetrachloromethane, toluene and tetrahydrofuran (THF) one or more.
Adopt the method for extraction agent oil and grease extracting relative to centrifugation, energy consumption can be reduced further.
In the present invention, oleaginous microorganism can be heterotrophic microorganism, can be also autotrophic microorganism, include but not limited to bacterium, fungi, actinomycetes and micro-algae, such as, can be schizochytrium limacinum, also known as splitting kettle algae.In the present invention, oleaginous microorganism is preferably micro-algae.Kind for micro-algae has no particular limits, be preferably oil-producing microalgae, be more preferably the produce oil engineering microalgae with larger industrial utilization, such as, micro-algae can be selected from least one in chlorella, grid algae, spirulina, chrysophyceae or Phaeodactylum tricornutum.
Embodiment
The present invention is further illustrated for following embodiment, but therefore do not limit the present invention.
In following embodiment and comparative example:
The aqueous slurry containing chlorella of 9 % by weight, 11 % by weight and 14 % by weight is mixed with the dry algae powder of chlorella and deionized water.
The weight of the weight/dry algae powder of the grease of grease extraction efficiency=extraction.
PH value is recorded by pH meter (Beijing Technew SA of Ge Le regular senior middle school, PH-9701).
Embodiment 1
This embodiment is for illustration of the method extracting grease from oleaginous microorganism of the present invention.
(1) take the aqueous slurry 45g containing chlorella of 11 % by weight, the pH value of the aqueous slurry regulated containing chlorella with sulfuric acid is 3;
(2) aqueous slurry containing chlorella is placed in mechanic whirl-nett reaction still, be under the hydrothermal condition of 200 DEG C in temperature, the material be stirred relative to every cubic metre, power of agitator is that 0.6kW(adopts Juancheng, Shandong Province Yongxing instrument plant YXMD type simulated reinforcement electric mixer to stir, lower same), stir 60 minutes, obtain the molten slurry of chlorella;
(3) by the centrifugation 5 minutes under 5000rpm of molten for chlorella slurry, collect upper oil phase and obtain grease 1.58g, calculating grease extraction efficiency is 31.9%.
Embodiment 2
This embodiment is for illustration of the method extracting grease from oleaginous microorganism of the present invention.
(1) take the aqueous slurry 35g containing chlorella of 14 % by weight, the pH value of the aqueous slurry regulated containing chlorella with sulfuric acid is 2;
(2) aqueous slurry containing chlorella being placed in mechanic whirl-nett reaction still, is under the hydrothermal condition of 260 DEG C in temperature, the material be stirred relative to every cubic metre, and power of agitator is 0.2kW, stirs 120 minutes, obtains the molten slurry of chlorella;
(3) by the centrifugation 6 minutes under 4000rpm of molten for chlorella slurry, collect upper oil phase and obtain grease 1.72g, calculating grease extraction efficiency is 35.1%.
Embodiment 3
This embodiment is for illustration of the method extracting grease from oleaginous microorganism of the present invention.
(1) take the aqueous slurry 55g containing chlorella of 9 % by weight, the pH value of the aqueous slurry regulated containing chlorella with sulfuric acid is 4;
(2) aqueous slurry containing chlorella being placed in mechanic whirl-nett reaction still, is under the hydrothermal condition of 160 DEG C in temperature, the material be stirred relative to every cubic metre, and power of agitator is 0.3kW, stirs 30 minutes, obtains the molten slurry of chlorella;
(3) by the centrifugation 4 minutes under 6000rpm of molten for chlorella slurry, collect upper oil phase and obtain grease 1.38g, calculating grease extraction efficiency is 27.9%.
Embodiment 4
This embodiment is for illustration of the method extracting grease from oleaginous microorganism of the present invention.
Grease is extracted from containing the aqueous slurry of chlorella according to the method for embodiment 1, unlike, in step (3), 30mL trichloromethane is joined in the molten slurry of chlorella, under agitation make the molten slurry of chlorella fully mix with trichloromethane, then stratification, collect extract layer, evaporate to dryness trichloromethane at 70 DEG C, obtains grease 1.6g, and calculating grease extraction efficiency is 32.3%.
Embodiment 5
This embodiment is for illustration of the method extracting grease from oleaginous microorganism of the present invention.
Grease is extracted from containing the aqueous slurry of chlorella according to the method for embodiment 2, unlike, in step (3), 49mL tetrachloromethane is joined in the molten slurry of chlorella, under agitation make the molten slurry of chlorella fully mix with tetrachloromethane, then stratification, collect extract layer, evaporate to dryness tetrachloromethane at 100 DEG C, obtains grease 1.73g, and calculating grease extraction efficiency is 35.3%.
Embodiment 6
This embodiment is for illustration of the method extracting grease from oleaginous microorganism of the present invention.
Grease is extracted from containing the aqueous slurry of chlorella according to the method for embodiment 3, unlike, in step (3), 40mL toluene is joined in the molten slurry of chlorella, under agitation make the molten slurry of chlorella fully mix with toluene, then stratification, collect extract layer, evaporate to dryness toluene at 120 DEG C, obtains grease 1.39g, and calculating grease extraction efficiency is 28.1%.
Embodiment 7
This embodiment is for illustration of the method extracting grease from oleaginous microorganism of the present invention.
Grease is extracted from containing the aqueous slurry of chlorella according to the method for embodiment 1, unlike, in step (1), the pH value of the aqueous slurry regulated containing chlorella with sulfuric acid is 1.Obtain grease 1.24g, calculating grease extraction efficiency is 25%.
Embodiment 8
This embodiment is for illustration of the method extracting grease from oleaginous microorganism of the present invention.
Grease is extracted from containing the aqueous slurry of chlorella according to the method for embodiment 1, unlike, in step (1), the pH value of the aqueous slurry regulated containing chlorella with sulfuric acid is 5.Obtain grease 1.27g, calculating grease extraction efficiency is 26%.
Comparative example 1
(1) take the aqueous slurry 45g containing chlorella of 11 % by weight, the pH value of the aqueous slurry regulated containing chlorella with NaOH is 8;
(2) aqueous slurry containing chlorella being placed in mechanic whirl-nett reaction still, is under the hydrothermal condition of 200 DEG C in temperature, the material be stirred relative to every cubic metre, and power of agitator is 0.6kW, stirs 60 minutes, obtains the molten slurry of chlorella;
(3) by the centrifugation 10 minutes under 5000rpm of molten for chlorella slurry, collect upper oil phase and obtain grease 0.05g, calculating grease extraction efficiency is 1%.
Embodiment 1 and comparative example 1 are compared and can be found out, the inventive method is compared with traditional hydrothermal method, and grease extraction efficiency is high.
Embodiment 1 and embodiment 4 are compared, embodiment 2 and embodiment 5 are compared, embodiment 3 and embodiment 6 are compared, can find out, adopt extraction agent oil and grease extracting, compared with centrifugation, grease extraction efficiency is more or less the same, and can reduce energy consumption further.
Embodiment 1 is compared can find out with embodiment 7 and embodiment 8 respectively, regulates aqueous slurry to the pH value containing oleaginous microorganism to be 2-4, be more conducive to improving grease extraction efficiency.
The method extracting grease from oleaginous microorganism provided by the invention, be the grease stripping made under the condition of 1-5 in oleaginous microorganism cell in pH value, aqueous phase and the oil phase of the molten slurry of the oleaginous microorganism obtained are easily separated, and grease extraction efficiency is high.
More than describe the preferred embodiment of the present invention in detail; but the present invention is not limited to the detail in above-mentioned embodiment, within the scope of technical conceive of the present invention; can carry out multiple simple variant to technical scheme of the present invention, these simple variant all belong to protection scope of the present invention.
It should be noted that in addition, each concrete technical characteristic described in above-mentioned embodiment, in reconcilable situation, can be combined by any suitable mode, in order to avoid unnecessary repetition, the present invention illustrates no longer separately to various possible array mode.
In addition, also can carry out arbitrary combination between various different embodiment of the present invention, as long as it is without prejudice to thought of the present invention, it should be considered as content disclosed in this invention equally.
Claims (13)
1. from oleaginous microorganism, extract a method for grease, it is characterized in that, described method comprises the steps:
(1) aqueous slurry to the pH value containing oleaginous microorganism is regulated to be 1-5;
(2) stir under hydrothermal conditions, make the grease stripping in oleaginous microorganism cell, obtain the molten slurry of oleaginous microorganism containing grease;
(3) from the molten slurry of oleaginous microorganism, grease is isolated.
2. method according to claim 1, wherein, in step (1), regulates aqueous slurry to the pH value containing oleaginous microorganism to be 2-4.
3. method according to claim 1 and 2, wherein, with the dry weight basis of oleaginous microorganism, described is 2-40 % by weight containing the content of oleaginous microorganism in the aqueous slurry of oleaginous microorganism.
4. method according to claim 1 and 2, wherein, in step (2), hydrothermal condition comprises: temperature is 120-400 DEG C.
5. method according to claim 4, wherein, in step (2), hydrothermal condition comprises: temperature is 160-260 DEG C.
6. method according to claim 1 and 2, wherein, in step (2), the condition of stirring comprises: the material be stirred relative to every cubic metre, and power of agitator is 0.15-1.5kW; Churning time is 2-250 minute.
7. method according to claim 6, wherein, in step (2), the condition of stirring comprises: the material be stirred relative to every cubic metre, and power of agitator is 0.2-1kW; Churning time is 30-120 minute.
8. method according to claim 1 and 2, wherein, in step (3), the method isolating grease from the molten slurry of oleaginous microorganism comprises centrifugation, and the condition of described centrifugation comprises: rotating speed 3000-8000rpm, time 2-10 minute.
9. method according to claim 1 and 2, wherein, in step (3), the method isolating grease from the molten slurry of oleaginous microorganism comprises and being contacted with extraction agent by molten for described oleaginous microorganism slurry, extracts grease.
10. method according to claim 9, wherein, with the dry weight basis of oleaginous microorganism, relative to 1g oleaginous microorganism, the consumption of described extraction agent is 5-20mL.
11. methods according to claim 9, wherein, described extraction agent be selected from hexane, methylene dichloride, trichloromethane, tetrachloromethane, toluene and tetrahydrofuran (THF) one or more.
12. methods according to claim 1 and 2, wherein, described oleaginous microorganism is micro-algae.
13. methods according to claim 12, wherein, described micro-algae is selected from least one in chlorella, grid algae, spirulina, chrysophyceae and Phaeodactylum tricornutum.
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| CN201310306509.4A CN104293473B (en) | 2013-07-19 | 2013-07-19 | A kind of method that oils and fatss are extracted from oleaginous microorganism |
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| CN201310306509.4A CN104293473B (en) | 2013-07-19 | 2013-07-19 | A kind of method that oils and fatss are extracted from oleaginous microorganism |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105080939A (en) * | 2015-09-07 | 2015-11-25 | 中国科学院烟台海岸带研究所 | Emergency rapid reduction method of ashore green tide algae |
| CN105505555A (en) * | 2016-01-28 | 2016-04-20 | 润科生物工程（福建）有限公司 | Method for extracting grease from fermentation liquor containing oil producing microbial cells |
| CN106221820A (en) * | 2016-07-21 | 2016-12-14 | 南京信息工程大学 | A kind of environment-friendly fuel and preparation method thereof |
| CN106701311A (en) * | 2016-12-28 | 2017-05-24 | 中国科学院南海海洋研究所 | Method for preparing microalgae oil and protein polysaccharide by adopting wet algae and one-step method |
-
2013
- 2013-07-19 CN CN201310306509.4A patent/CN104293473B/en active Active
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105080939A (en) * | 2015-09-07 | 2015-11-25 | 中国科学院烟台海岸带研究所 | Emergency rapid reduction method of ashore green tide algae |
| CN105505555A (en) * | 2016-01-28 | 2016-04-20 | 润科生物工程（福建）有限公司 | Method for extracting grease from fermentation liquor containing oil producing microbial cells |
| CN105505555B (en) * | 2016-01-28 | 2020-01-03 | 润科生物工程（福建）有限公司 | Method for extracting grease from fermentation liquor containing oil-producing microbial cells |
| CN106221820A (en) * | 2016-07-21 | 2016-12-14 | 南京信息工程大学 | A kind of environment-friendly fuel and preparation method thereof |
| CN106701311A (en) * | 2016-12-28 | 2017-05-24 | 中国科学院南海海洋研究所 | Method for preparing microalgae oil and protein polysaccharide by adopting wet algae and one-step method |
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| CN104293473B (en) | 2017-03-29 |
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