CN104293473B - A kind of method that oils and fatss are extracted from oleaginous microorganism - Google Patents
A kind of method that oils and fatss are extracted from oleaginous microorganism Download PDFInfo
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- CN104293473B CN104293473B CN201310306509.4A CN201310306509A CN104293473B CN 104293473 B CN104293473 B CN 104293473B CN 201310306509 A CN201310306509 A CN 201310306509A CN 104293473 B CN104293473 B CN 104293473B
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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
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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
- C11B1/10—Production of fats or fatty oils from raw materials by extracting
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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
- 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 kind of method that oils and fatss are extracted from oleaginous microorganism, it is characterised in that methods described comprises the steps：（1）It is 15 to adjust the aqueous slurry containing oleaginous microorganism to pH value；（2）Stir under hydrothermal conditions, make the oils and fatss dissolution in oleaginous microorganism cell, obtain the molten slurry of the oleaginous microorganism containing oils and fatss；（3）Oils and fatss are isolated from the molten slurry of oleaginous microorganism.The method that oils and fatss are extracted from oleaginous microorganism that the present invention is provided, is the oils and fatss dissolution in making oleaginous microorganism cell under conditions of 15 in pH value, and the water phase and oil phase of the molten slurry of the oleaginous microorganism for obtaining is easily separated, and grease extraction efficiency is high.
Description
Technical field
The present invention relates to oleaginous microorganism field of engineering technology, in particular it relates to one kind extract oil from oleaginous microorganism
The method of fat.
Background technology
Due to microbial cell propagation soon, with short production cycle, growth of microorganism needed raw material wide material sources, low price,
And growth of microorganism is not limited by season, climate change, the continuous large-scale production of energy, low production cost, and oleaginous microorganism
Middle oil content is high, therefore, oils and fatss are produced using oleaginous microorganism have become a kind of important means for alleviating global energy crisis.
Microalgae is a kind of aquatic phototroph, and it utilizes luminous energy, with CO2And H2O carries out the life of cell as substrate
It is long, and synthesize substantial amounts of triglyceride in cell（TAGs）, polysaccharide, the material such as protein.Wherein, triglyceride（TAGs）Bag
Include unsaturated glyceride（DHA, EPA）Not only can be used as important nutrient and healthcare products, and can be used as biological liquid fuel
Important glyceride stock source.In particular with the day by day shortage of world energy sources such as oil, with animal and plant fat（Including microalgae oil
Fat）Receive significant attention for raw material production bio-fuel, and competitively carry out the research of microalgae biodiesel.
The extraction of microalgae grease is always the key technology of biological diesel oil refining.The microalgae of most of species has cellulose
Sexual cell wall, some kinds of microalgae cell wall are also difficult to the other compositions of digest and decompose in a large number, and cell wall thickness, quality are hard
It is tough, the extraction of intracellular oils and fatss and nutrient substance is highly detrimental to, and needs broken wall treatment to be carried out to microalgae.
The extraction of microalgae grease at present generally adopts hydro-thermal method.Hydro-thermal method typically passes through High Temperature High Pressure in the basic conditions
Cell wall breaking, oils and fatss dissolution are carried out, general temperature is 100-300 DEG C, the molten slurry of gained microalgae carries out oil-water separation by centrifugation, such as
CN100460513C, but as the molten slurry of the microalgae for obtaining is the emulsion containing algae-residue, it is uniform that oil phase is mixed with water conjunction, it is difficult to point
From grease extraction efficiency is relatively low.
Therefore, the efficient and economic technology that oils and fatss are extracted from oleaginous microorganism becomes the emphasis of research.
The content of the invention
The purpose of the present invention is to overcome in prior art the low defect of grease extraction efficiency is extracted from oleaginous microorganism, there is provided
A kind of efficient method that oils and fatss are extracted from oleaginous microorganism.
The present inventor has been surprisingly found that under study for action, in making oleaginous microorganism cell under conditions of pH value is 1-5
Oils and fatss dissolution, the water phase and oil phase of the molten slurry of the oleaginous microorganism for obtaining be easily separated, and grease extraction efficiency is high.
Therefore, to achieve these goals, the invention provides it is a kind of from oleaginous microorganism extract oils and fatss method, its
It is characterised by, methods described comprises the steps：
（1）It is 1-5 to adjust the aqueous slurry containing oleaginous microorganism to pH value；
（2）Stir under hydrothermal conditions, make the oils and fatss dissolution in oleaginous microorganism cell, obtain the oil-producing containing oils and fatss micro-
Biological molten slurry；
（3）Oils and fatss are isolated from the molten slurry of oleaginous microorganism.
Preferably, step（1）In, it is 2-4 to adjust the aqueous slurry containing oleaginous microorganism to pH value.
Preferably, in terms of the dry weight of oleaginous microorganism, oleaginous microorganism in the aqueous slurry containing oleaginous microorganism
Content is 2-40 weight %.
Preferably, step（2）In, hydrothermal condition includes：Temperature is 120-400 DEG C, preferably 160-260 DEG C.
Preferably, step（2）In, the condition of stirring includes：Relative to per cubic metre of material being stirred, power of agitator is
0.15-1.5kW, preferably 0.2-1kW；Mixing time is 2-250 minutes, preferably 30-120 minutes.
Preferably, step（3）In, the method that oils and fatss are isolated from the molten slurry of oleaginous microorganism includes centrifugation, described
The condition of centrifugation includes：Rotating speed 3000-8000rpm, minute time 2-10.
Preferably, step（3）In, the method that oils and fatss are isolated from the molten slurry of oleaginous microorganism is included the oil-producing micro- life
The molten slurry of thing is contacted with extractant, extracts oils and fatss.
Preferably, in terms of the dry weight of oleaginous microorganism, relative to 1g oleaginous microorganisms, the consumption of the extractant is 5-
20mL。
Preferably, the extractant is in hexane, dichloromethane, chloroform, tetrachloromethane, toluene and tetrahydrofuran
One or more.
Preferably, the oleaginous microorganism is microalgae.The microalgae be preferably selected from chlorella, scenedesmus, spirulina, chrysophyceae and
At least one in Phaeodactylum.
The method that oils and fatss are extracted from oleaginous microorganism that the present invention is provided, makes oil-producing micro- under conditions of pH value is 1-5
Oils and fatss dissolution in biological cell, the water phase and oil phase of the molten slurry of the oleaginous microorganism for obtaining are easily separated, and grease extraction efficiency is high.
Other features and advantages of the present invention will be described in detail in subsequent specific embodiment part.
Specific embodiment
Hereinafter the specific embodiment of the present invention is described in detail.It should be appreciated that described herein concrete
Embodiment is merely to illustrate and explains the present invention, is not limited to the present invention.
The invention provides a kind of method that oils and fatss are extracted from oleaginous microorganism, the method comprises the steps：
（1）It is 1-5 to adjust the aqueous slurry containing oleaginous microorganism to pH value；
（2）Stir under hydrothermal conditions, make the oils and fatss dissolution in oleaginous microorganism cell, obtain the oil-producing containing oils and fatss micro-
Biological molten slurry；
（3）Oils and fatss are isolated from the molten slurry of oleaginous microorganism.
According to the present invention, the purpose of the present invention is capable of achieving although with said method, even if the molten slurry of oleaginous microorganism
Water phase and oil phase are easily separated, and grease extraction efficiency is high.But under preferable case, step（1）In, adjust the water slurry containing oleaginous microorganism
Liquid is 2-4 to pH value, can further improve grease extraction efficiency.
In the present invention, the aqueous slurry containing oleaginous microorganism is referred to collects oleaginous microorganism from oleaginous microorganism culture fluid
The liquid containing oleaginous microorganism for obtaining.In terms of the dry weight of oleaginous microorganism, in the aqueous slurry containing oleaginous microorganism, oil-producing is micro-
Biological content is preferably 2-40 weight %, more preferably 7-15 weight %.
In the present invention, for pH value condition, can be adjusted with acid adding, for acid is without particular/special requirement, ability can be adopted
The conventional acid in domain, for example, acid can be selected from sulphuric acid, hydrochloric acid, formic acid, acetic acid, citric acid, lactic acid, maleic acid, tartaric acid and oxalic acid
In one or more.
In the present invention, pH value is measured by pH meter.
In the present invention, hydrothermal condition refers to the condition for keeping liquid at a certain temperature.Step of the present invention（2）In, hydro-thermal
Condition is preferably included：Temperature is 120-400 DEG C；More preferably include：Temperature is 160-260 DEG C.
It will be understood by those skilled in the art that step（2）In, as long as the pressure of hydrothermal condition is allowed the water in above-mentioned temperature
Degree is lower to keep liquid.
Step of the present invention（2）In, for the condition of stirring, as long as the oils and fatss dissolution in making oleaginous microorganism cell.
For example, the condition of stirring can include：Relative to per cubic metre of material being stirred, power of agitator is 0.15-1.5kW；Stirring
Time is 2-250 minutes；Preferably, the condition of stirring can include：Relative to per cubic metre of material being stirred, power of agitator
For 0.2-1kW；Mixing time is 30-120 minutes.Under above-mentioned stirring condition, the oils and fatss in oleaginous microorganism cell can be made
Dissolution.
In the present invention, the molten slurry of oleaginous microorganism refers to oleaginous microorganism cell wall in the aqueous slurry containing oleaginous microorganism
Destruction, intracellular structure are disintegrated, the mixture that oleaginous microorganism oils and fatss dissolution is obtained.
Step of the present invention（3）In, oils and fatss are isolated from the molten slurry of oleaginous microorganism, can be extracted in oils and fatss using hydro-thermal method
Conventional method, such as centrifugation, but the water phase and oil phase of the molten slurry of oleaginous microorganism obtained due to the inventive method easily divided
From, accordingly, with respect to traditional hydro-thermal method, the energy consumption of centrifugation can be reduced, the condition of centrifugation is preferably included：Rotating speed
3000-8000rpm, minute time 2-10；More preferably include：Rotating speed 4000-6000rpm, minute time 4-6.
The water phase and oil phase of the molten slurry of oleaginous microorganism obtained due to the inventive method is easily separated, therefore, except traditional water
Outside centrifugal separation method used in full-boiled process, the method that oils and fatss are isolated from the molten slurry of oleaginous microorganism can also be by oil-producing micro- life
The molten slurry of thing is contacted with extractant, extracts oils and fatss.
In the present invention, oleaginous microorganism molten slurry is contacted with extractant, the method for extracting oils and fatss is by oil-producing micro- life
The molten slurry of thing is contacted with extractant, is sufficiently mixed, then stratification, is collected extract layer, using extractant boiling point less than oils and fatss
Property, is evaporated extractant, so as to obtain oils and fatss.
In the present invention, in terms of the dry weight of oleaginous microorganism, relative to 1g oleaginous microorganisms, the consumption of extractant is preferably 5-
20mL；More preferably 5-10mL.
In the present invention, for extractant is without particular/special requirement, the extractant that oil and grease extracting can be adopted to commonly use, for example, extract
Agent can be selected from one or more in hexane, dichloromethane, chloroform, tetrachloromethane, toluene and tetrahydrofuran.
Using the method for extractant oil and grease extracting relative to centrifugation, can further reducing energy consumption.
In the present invention, oleaginous microorganism can be heterotrophic microorganism, or autotrophic microbe, including but not limited to carefully
Bacterium, funguses, actinomycetes and microalgae, for example, can be schizochytrium limacinum, also known as splitting kettle algae.In the present invention, oleaginous microorganism is preferably
Microalgae.For the species of microalgae has no particular limits, preferably oil-producing microalgae, more preferably with larger industry exploitation value
The oil-producing engineering microalgae of value, for example, microalgae can be in chlorella, scenedesmus, spirulina, chrysophyceae or Phaeodactylum extremely
Few one kind.
Embodiment
The present invention is further illustrated for below example, but and is not so limited the present invention.
In following embodiments and comparative example：
With the dry algae powder and deionized water of chlorella be configured to 9 weight %, 11 weight % and 14 weight % containing chlorella
Aqueous slurry.
The weight of the weight/dry algae powder of the oils and fatss of grease extraction efficiency=extraction.
PH value passes through pH meter（Beijing Technew SA of Ge Le regular senior middle schools, PH-9701）Measure.
Embodiment 1
The embodiment is used for the method that oils and fatss are extracted from oleaginous microorganism for illustrating the present invention.
（1）Aqueous slurry 45g containing chlorella of 11 weight % is weighed, and the aqueous slurry containing chlorella is adjusted with sulphuric acid
PH value is 3；
（2）Aqueous slurry containing chlorella is placed in mechanic whirl-nett reaction kettle, in the hydrothermal condition that temperature is 200 DEG C
Under, relative to per cubic metre of material being stirred, power of agitator is 0.6kW（Using Shandong Province Juancheng Yongxing instrument plant YXMD types
Simulation reinforcement motor stirrer is stirred, similarly hereinafter）, stir 60 minutes, obtain the molten slurry of chlorella；
（3）By chlorella molten slurry centrifugation 5 minutes under 5000rpm, collect upper oil phase and obtain oils and fatss 1.58g, count
It is 31.9% to calculate grease extraction efficiency.
Embodiment 2
The embodiment is used for the method that oils and fatss are extracted from oleaginous microorganism for illustrating the present invention.
（1）Aqueous slurry 35g containing chlorella of 14 weight % is weighed, and the aqueous slurry containing chlorella is adjusted with sulphuric acid
PH value is 2；
（2）Aqueous slurry containing chlorella is placed in mechanic whirl-nett reaction kettle, in the hydrothermal condition that temperature is 260 DEG C
Under, relative to per cubic metre of material being stirred, power of agitator is 0.2kW, stirs 120 minutes, obtains the molten slurry of chlorella；
（3）By chlorella molten slurry centrifugation 6 minutes under 4000rpm, collect upper oil phase and obtain oils and fatss 1.72g, count
It is 35.1% to calculate grease extraction efficiency.
Embodiment 3
The embodiment is used for the method that oils and fatss are extracted from oleaginous microorganism for illustrating the present invention.
（1）Aqueous slurry 55g containing chlorella of 9 weight % is weighed, and the pH of the aqueous slurry containing chlorella is adjusted with sulphuric acid
It is worth for 4；
（2）Aqueous slurry containing chlorella is placed in mechanic whirl-nett reaction kettle, in the hydrothermal condition that temperature is 160 DEG C
Under, relative to per cubic metre of material being stirred, power of agitator is 0.3kW, stirs 30 minutes, obtains the molten slurry of chlorella；
（3）By chlorella molten slurry centrifugation 4 minutes under 6000rpm, collect upper oil phase and obtain oils and fatss 1.38g, count
It is 27.9% to calculate grease extraction efficiency.
Embodiment 4
The embodiment is used for the method that oils and fatss are extracted from oleaginous microorganism for illustrating the present invention.
Oils and fatss are extracted from the aqueous slurry containing chlorella according to the method for embodiment 1, except for the difference that, in step（3）In,
30mL chloroform is added in the molten slurry of chlorella, the molten slurry of chlorella is sufficiently mixed with chloroform under agitation, then
Stratification, collects extract layer, is evaporated chloroform at 70 DEG C, obtains oils and fatss 1.6g, and it is 32.3% to calculate grease extraction efficiency.
Embodiment 5
The embodiment is used for the method that oils and fatss are extracted from oleaginous microorganism for illustrating the present invention.
Oils and fatss are extracted from the aqueous slurry containing chlorella according to the method for embodiment 2, except for the difference that, in step（3）In,
49mL tetrachloromethanes are added in the molten slurry of chlorella, the molten slurry of chlorella is sufficiently mixed with tetrachloromethane under agitation, then
Stratification, collects extract layer, is evaporated tetrachloromethane at 100 DEG C, obtains oils and fatss 1.73g, calculates grease extraction efficiency and is
35.3%。
Embodiment 6
The embodiment is used for the method that oils and fatss are extracted from oleaginous microorganism for illustrating the present invention.
Oils and fatss are extracted from the aqueous slurry containing chlorella according to the method for embodiment 3, except for the difference that, in step（3）In,
40mL toluene is added in the molten slurry of chlorella, the molten slurry of chlorella is sufficiently mixed with toluene under agitation, then stratification,
Extract layer being collected, toluene being evaporated at 120 DEG C, obtain oils and fatss 1.39g, it is 28.1% to calculate grease extraction efficiency.
Embodiment 7
The embodiment is used for the method that oils and fatss are extracted from oleaginous microorganism for illustrating the present invention.
Oils and fatss, except for the difference that, step are extracted from the aqueous slurry containing chlorella according to the method for embodiment 1（1）In, use
It is 1 that sulphuric acid adjusts the pH value of the aqueous slurry containing chlorella.Oils and fatss 1.24g is obtained, it is 25% to calculate grease extraction efficiency.
Embodiment 8
The embodiment is used for the method that oils and fatss are extracted from oleaginous microorganism for illustrating the present invention.
Oils and fatss, except for the difference that, step are extracted from the aqueous slurry containing chlorella according to the method for embodiment 1（1）In, use
It is 5 that sulphuric acid adjusts the pH value of the aqueous slurry containing chlorella.Oils and fatss 1.27g is obtained, it is 26% to calculate grease extraction efficiency.
Comparative example 1
（1）Aqueous slurry 45g containing chlorella of 11 weight % is weighed, and the aqueous slurry containing chlorella is adjusted with NaOH
PH value is 8；
（2）Aqueous slurry containing chlorella is placed in mechanic whirl-nett reaction kettle, in the hydrothermal condition that temperature is 200 DEG C
Under, relative to per cubic metre of material being stirred, power of agitator is 0.6kW, stirs 60 minutes, obtains the molten slurry of chlorella；
（3）By chlorella molten slurry centrifugation 10 minutes under 5000rpm, collect upper oil phase and obtain oils and fatss 0.05g, count
It is 1% to calculate grease extraction efficiency.
By embodiment 1 with comparative example 1 be compared as can be seen that the inventive method compared with traditional hydro-thermal method, oils and fatss
Extraction ratio is high.
Embodiment 1 is compared with embodiment 4, embodiment 2 is compared with embodiment 5, by embodiment 3 and enforcement
Example 6 is compared, it can be seen that using extractant oil and grease extracting, compared with centrifugation, grease extraction efficiency is more or less the same, and
Can further reducing energy consumption.
Embodiment 1 is compared with embodiment 7 and embodiment 8 respectively as can be seen that adjusting containing oleaginous microorganism
Aqueous slurry is 2-4 to pH value, is more beneficial for improving grease extraction efficiency.
The method that oils and fatss are extracted from oleaginous microorganism that the present invention is provided, makes oil-producing micro- under conditions of pH value is 1-5
Oils and fatss dissolution in biological cell, the water phase and oil phase of the molten slurry of the oleaginous microorganism for obtaining are easily separated, and grease extraction efficiency is high.
The preferred embodiment of the present invention described in detail above, but, the present invention is not limited in above-mentioned embodiment
Detail, the present invention range of the technology design in, various simple variants can be carried out to technical scheme, this
A little simple variants belong to protection scope of the present invention.
It is further to note that each particular technique feature described in above-mentioned specific embodiment, in not lance
In the case of shield, can be combined by any suitable means, in order to avoid unnecessary repetition, the present invention to it is various can
The compound mode of energy is no longer separately illustrated.
Additionally, combination in any between a variety of embodiments of the present invention, can also be carried out, as long as which is without prejudice to this
The thought of invention, which should equally be considered as content disclosed in this invention.
Claims (10)
1. it is a kind of from oleaginous microorganism extract oils and fatss method, it is characterised in that methods described comprises the steps：
(1) it is 1-5 to adjust the aqueous slurry containing oleaginous microorganism to pH value；
(2) stir under hydrothermal conditions, make the oils and fatss dissolution in oleaginous microorganism cell, obtain the oleaginous microorganism containing oils and fatss
Molten slurry；
(3) oils and fatss are isolated from the molten slurry of oleaginous microorganism；
In step (2), hydrothermal condition includes：Temperature is 120-400 DEG C；
At least one of the oleaginous microorganism in chlorella, scenedesmus, spirulina, chrysophyceae and Phaeodactylum.
2. method according to claim 1, wherein, in step (1), the aqueous slurry containing oleaginous microorganism is adjusted to pH value
For 2-4.
3. method according to claim 1 and 2, wherein, it is in terms of the dry weight of oleaginous microorganism, described to contain oleaginous microorganism
Aqueous slurry in oleaginous microorganism content be 2-40 weight %.
4. method according to claim 1, wherein, in step (2), hydrothermal condition includes：Temperature is 160-260 DEG C.
5. method according to claim 1, wherein, in step (2), the condition of stirring includes：Relative to per cubic metre of quilt
The material of stirring, power of agitator are 0.15-1.5kW；Mixing time is 2-250 minutes.
6. method according to claim 5, wherein, in step (2), the condition of stirring includes：Relative to per cubic metre of quilt
The material of stirring, power of agitator are 0.2-1kW；Mixing time is 30-120 minutes.
7. method according to claim 1 and 2, wherein, in step (3), oils and fatss are isolated from the molten slurry of oleaginous microorganism
Method include centrifugation, the condition of the centrifugation includes：Rotating speed 3000-8000rpm, minute time 2-10.
8. method according to claim 1 and 2, wherein, in step (3), oils and fatss are isolated from the molten slurry of oleaginous microorganism
Method include by it is the oleaginous microorganism molten slurry contact with extractant, extract oils and fatss.
9. method according to claim 8, wherein, in terms of the dry weight of oleaginous microorganism, relative to 1g oleaginous microorganisms, institute
The consumption for stating extractant is 5-20mL.
10. method according to claim 8, wherein, the extractant is selected from hexane, dichloromethane, chloroform, tetrachloro
One or more in methane, toluene and tetrahydrofuran.
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| 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 |
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