CN109022141A - A method of extracting microbial oil - Google Patents
A method of extracting microbial oil Download PDFInfo
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- CN109022141A CN109022141A CN201811157869.1A CN201811157869A CN109022141A CN 109022141 A CN109022141 A CN 109022141A CN 201811157869 A CN201811157869 A CN 201811157869A CN 109022141 A CN109022141 A CN 109022141A
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- Prior art keywords
- microbial oil
- oil
- isopropanol
- added
- extracted
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Classifications
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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
-
- 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
- C11B1/108—Production of fats or fatty oils from raw materials by extracting after-treatment, e.g. of miscellae
Abstract
The present invention provides a kind of method for extracting microbial oil, and the method continuously extracted by low temperature avoids high temperature from consuming grease, and low energy consumption, and the adjustment of the technical parameters such as change of gradient for combining pH, realizes that the good high quality of rhodotorula glutinis is extracted.It is up to 98.4% using the recovery rate of the method for the present invention microbial oil, impurity content can be down to 0 in gained microbial oil, and for unsaponifiable matter content down to 0.2%, oil quality is good.
Description
Technical field
The present invention relates to technical field of oil extraction, in particular to extract the method for microbial oil.
Background technique
What preparation biodiesel mainly utilized at present is the grease extracted in vegetable oil or castoff of food and drink, vegetable oil raw materials
High production cost, it is inconvenient that two castoff of food and drink are collected, and centralized production has certain difficulty.Microbial reproduction and metabolism period are short
And fat content is high, compared with vegetable oil, is not take up arable land, can continuous production, not climate and seasonal effect.It is as substitution
Oil sources is environmentally friendly, and reproducibility is strong, is the research hotspot of various countries technical staff.
Oleaginous microorganism is because of the substitution energy that a series of advantages of its own are considered as most promising production biodiesel
Source.For rhodotorula glutinis, growth cycle is short, and fat content is abundant, generally can achieve the 40%~70% of dry cell weight,
The oil production of unit area can be higher by the oil crops decades of times such as soybean.
Nevertheless, higher cost is still that microbial oil industry realizes industrialized bottleneck, among these, the extraction of grease
Cost accounts for about the 50% of totle drilling cost.Currently, microbial oil extractive technique has Mechanical Method, enzyme process and organic solvent method.Mechanical Method
There is a problem of that grease extraction efficiency is relatively low, energy consumption is higher；Enzyme process is although efficient and environmental-friendly, but cost it is excessively high hinder its into
The application of one step；Organic solvent method generallys use chloroform, methanol, ether etc. and extracts grease, organic solvent from oleaginous microorganism
Dosage is big, extraction time is long, environment friendly is poor, and industrial applications are limited.
CN104531342A discloses a kind of method for extracting microbial oil, comprising the following steps: (1) will be after fermentation
Oleaginous microorganism obtains the wet thallus that water content is 65%~90% by separation system；(2) into the wet thallus of step (1)
The mixed solvent of n-hexane and ethyl alcohol is added, constant temperature extracts a period of time, is then separated by solid-liquid separation, obtains oil-containing mixed solvent
A and the bacterium dregs of rice；(3) n-hexane and certain density ethyl alcohol are added into the bacterium dregs of rice obtained by step (2), constant temperature extracts a period of time, then
Oil-containing mixed solvent b and the bacterium dregs of rice are collected after secondary separation of solid and liquid；(4) merge above-mentioned oil-containing mixed solvent a and b；Then precipitation obtains
Microbial oil.The n-hexane used in this method still has certain hypotoxicity, and product impurity content is more, and through excessive
Secondary separation of solid and liquid leads to the loss of grease, furthermore operates relatively complicated, has some limitations in popularization and application scheme.
Summary of the invention
Mirror is with this, and the present invention provides a kind of new methods for extracting microbial oil.
The technical scheme is that
A method of extracting microbial oil, comprising the following steps:
(1) wet thallus of rhodotorula glutinis is taken, the ethanol water that mass percentage concentration is 70~85% is added, adjusts pH
1~1.5h are extracted in 8.0~8.4,5~10 DEG C of concussions, and keep after improving temperature in 5~10min to 25~30 DEG C 20~
30min；
(2) isopropanol alcohol mixed solution is added, 10~30min is extracted in 15~20 DEG C of concussions；0.5~1 times of volume is added
Ethyl alcohol adjusts pH 3.0~5.5, continuously stirs 15~20min；Wherein, the additional amount of isopropanol alcohol mixed solution is step
(1) 2~3 times of gained feed liquid total volume；
(3) tetrahydrofuran is added, is separated by solid-liquid separation and is contained after 2.0~4.0, -5~0 DEG C of 1~2h of concussion extraction of adjusting pH
Oily mixed solvent；Wherein, the additional amount of tetrahydrofuran is 2~3 times of feed liquid total volume obtained by step (2)；
(4) vacuum desolvation under the conditions of being not higher than 50 DEG C of temperature, then conventional decoloration, deodorization processing, obtain microbial oil
Rouge.
Preferably, in step (2), pH 3.5.
Preferably, in step (2), the volume ratio of isopropanol and ethyl alcohol is 1:(7~10 in isopropanol alcohol mixed solution).
It is furthermore preferred that the volume ratio of isopropanol and ethyl alcohol is 1:8 in isopropanol alcohol mixed solution in step (2).
Preferably, the solid-liquid ratio g/mL of wet thallus and ethanol water is 1:(3~5 in step (1)).
It is furthermore preferred that the solid-liquid ratio g/mL of wet thallus and ethanol water is 1:3 in step (1).
Preferably, the water content of wet thallus is 50% weight ratio.
Preferably, step (3) are as follows: tetrahydrofuran is added, adjusts pH 2.2, is separated by solid-liquid separation and obtains after -5 DEG C of concussions extraction 1h
Oil-containing mixed solvent；Wherein, the additional amount of tetrahydrofuran is 2 times of feed liquid total volume obtained by step (2).
Compared with prior art, the beneficial effects of the present invention are:
A. the method that the present invention is continuously extracted by low temperature avoids high temperature from consuming grease, and low energy consumption, and combines the gradient of pH
The adjustment of the technical parameters such as variation realizes that the good high quality of rhodotorula glutinis is extracted.
B. the present invention belongs to the organic solvent of micro- poison using ethyl alcohol, petroleum ether, tetrahydrofuran and isopropanol, avoids to people
The harm of body, and it is conducive to industrial application.
C. rhodotorula glutinis thallus individual is big, and metabolin is more, itself protein rich in and a large amount of pigments are red from gluing
Extract high quality, the health of the high yield pulp1 difficulty certain with the presence of grease in yeast, the method for the present invention for rhodotorula glutinis and
Design, is up to 98.4% using the recovery rate of the method for the present invention microbial oil, impurity content in gained microbial oil
Can be down to 0, for unsaponifiable matter content down to 0.2%, oil quality is good.
Specific embodiment
In order to be best understood from the technology of the present invention content, specific embodiment is provided below, the present invention is described further.
Embodiment 1
A method of extracting microbial oil, comprising the following steps:
(1) it takes the wet thallus of rhodotorula glutinis, the ethanol water that mass percentage concentration is 85% is added, adjust pH8.4,10
1.5h is extracted in DEG C concussion, and keeps 30min after improving temperature in 5~10min to 30 DEG C；
(2) isopropanol alcohol mixed solution is added, 30min is extracted in 20 DEG C of concussions；1 times of volume ethanol is added, adjusts pH
5.5, continuously stir 20min；Wherein, the additional amount of isopropanol alcohol mixed solution is 3 of feed liquid total volume obtained by step (1)
Times；
(3) tetrahydrofuran is added, adjusts pH4.0, separation of solid and liquid obtains oil-containing mixed solvent after 2h is extracted in 0 DEG C of concussion；Its
In, the additional amount of tetrahydrofuran is 3 times of feed liquid total volume obtained by step (2)；
(4) vacuum desolvation under the conditions of being not higher than 50 DEG C of temperature, then conventional decoloration, deodorization processing, obtain microbial oil
Rouge.
In step (2), the volume ratio of isopropanol and ethyl alcohol is 1:6 in isopropanol alcohol mixed solution.
The solid-liquid ratio g/mL of wet thallus and ethanol water is 1:2 in step (1).
The water content of wet thallus is 70% weight ratio.
Embodiment 2
A method of extracting microbial oil, comprising the following steps:
(1) it takes the wet thallus of rhodotorula glutinis, the ethanol water that mass percentage concentration is 70% is added, adjust pH8.0,5
1h is extracted in DEG C concussion, and keeps 20min after improving temperature in 5~10min to 25 DEG C；
(2) isopropanol alcohol mixed solution is added, 10min is extracted in 15 DEG C of concussions；0.5 times of volume ethanol is added, adjusts pH
3.0, continuously stir 15min；Wherein, the additional amount of isopropanol alcohol mixed solution is 2 of feed liquid total volume obtained by step (1)
Times；
(3) tetrahydrofuran is added, adjusts pH 2.0, separation of solid and liquid obtains oil-containing mixed solvent after 1h is extracted in -5 DEG C of concussions；
Wherein, the additional amount of tetrahydrofuran is 2 times of feed liquid total volume obtained by step (2)；
(4) vacuum desolvation under the conditions of being not higher than 50 DEG C of temperature, then conventional decoloration, deodorization processing, obtain microbial oil
Rouge.
In step (2), the volume ratio of isopropanol and ethyl alcohol is 1:6 in isopropanol alcohol mixed solution.
The solid-liquid ratio g/mL of wet thallus and ethanol water is 1:2 in step (1).
The water content of wet thallus is 40% weight ratio.
Embodiment 3
A method of extracting microbial oil, comprising the following steps:
(1) it takes the wet thallus of rhodotorula glutinis, the ethanol water that mass percentage concentration is 70% is added, adjust pH8.0,6
1h is extracted in DEG C concussion, and keeps 25min after improving temperature in 5~10min to 25 DEG C；
(2) the isopropanol alcohol mixed solution of 2 times of volumes is added, 30min is extracted in 15 DEG C of concussions；0.5 times of volume second is added
Alcohol adjusts pH 3.5, continuously stirs 20min；Wherein, the additional amount of isopropanol alcohol mixed solution is feed liquid obtained by step (1)
2 times of total volume；
(3) tetrahydrofuran is added, adjusts pH 2.2, separation of solid and liquid obtains oil-containing mixed solvent after 1h is extracted in -5 DEG C of concussions；
Wherein, the additional amount of tetrahydrofuran is 2 times of feed liquid total volume obtained by step (2)；
(4) vacuum desolvation under the conditions of being not higher than 50 DEG C of temperature, then conventional decoloration, deodorization processing, obtain microbial oil
Rouge.
In step (2), the volume ratio of isopropanol and ethyl alcohol is 1:8 in isopropanol alcohol mixed solution.
The solid-liquid ratio g/mL of wet thallus and ethanol water is 1:3 in step (1).
The water content of wet thallus is 50% weight ratio.
Embodiment 4
A method of microbial oil is extracted, the difference with embodiment 3 is,
In step (2), the volume ratio of isopropanol and ethyl alcohol is 1:7 in isopropanol alcohol mixed solution.
Embodiment 5
A method of microbial oil is extracted, the difference with embodiment 3 is,
In step (2), the volume ratio of isopropanol and ethyl alcohol is 1:10 in isopropanol alcohol mixed solution
Embodiment 6
A method of microbial oil is extracted, the difference with embodiment 3 is,
The solid-liquid ratio g/mL of wet thallus and ethanol water is 1:5 in step (1).
Comparative example 1
A method of microbial oil is extracted, the difference from embodiment 1 is that, rhodotorula glutinis is replaced with into chlorella.
Comparative example 2
A method of extracting microbial oil, comprising the following steps:
The wet thallus for weighing the rhodotorula glutinis that weight in wet base is 2.0g (water content 80.0%) is added the n-hexane of 8mL and divides
It analyses straight alcohol mixed liquor (volume ratio 1:2.2), after 30 DEG C, 220rpm mechanical shaking extraction 3 hours, is centrifuged 10min under 8000rpm,
Collect oil-containing mixed solvent.75% ethyl alcohol and n-hexane mixed solvent (volume of 10mL are added into the bacterium dregs of rice after primary extract
Than after 30 DEG C, 220rpm mechanical shaking extraction 2 hours, 10min being centrifuged under 8000rpm, collects oil-containing mixed solvent for 3:1).To
75% ethyl alcohol and n-hexane mixed solvent (volume ratio 3:1) of 10mL, 30 DEG C, 220rpm are added in the bacterium dregs of rice after second extraction
After mechanical shaking extraction 1 hour, it is centrifuged 10min under 8000rpm, collects oil-containing mixed solvent.It is taken off after merging oil-containing mixed solvent three times
Molten, then conventional decoloration, deodorization processing, obtain microbial oil.
Microbial oil is extracted according to the method for embodiment and comparative example respectively, detects grease extraction efficiency and impurity content,
It the results are shown in Table 1.
Table 1
As known from Table 1, it is up to 98.4% using the recovery rate of the method for the present invention microbial oil, gained microbial oil
Impurity content can be down to 0 in rouge, and for unsaponifiable matter content down to 0.2%, oil quality is good.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Within mind and principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.
Claims (8)
1. a kind of method for extracting microbial oil, which comprises the following steps:
(1) take the wet thallus of rhodotorula glutinis, the ethanol water that mass percentage concentration is 70~85% be added, adjust pH 8.0~
1~1.5h is extracted in 8.4,5~10 DEG C of concussions, and 20~30min is kept after improving temperature in 5~10min to 25~30 DEG C；
(2) isopropanol alcohol mixed solution is added, 10~30min is extracted in 15~20 DEG C of concussions；0.5~1 times of volume ethanol is added,
PH 3.0~5.5 is adjusted, 15~20min is continuously stirred；Wherein, the additional amount of isopropanol alcohol mixed solution is step (1) institute
Obtain 2~3 times of feed liquid total volume；
(3) tetrahydrofuran is added, it is mixed to obtain oil-containing for separation of solid and liquid after 2.0~4.0, -5~0 DEG C of 1~2h of concussion extraction of adjusting pH
Bonding solvent；Wherein, the additional amount of tetrahydrofuran is 2~3 times of feed liquid total volume obtained by step (2)；
(4) vacuum desolvation under the conditions of being not higher than 50 DEG C of temperature, then conventional decoloration, deodorization processing, obtain microbial oil.
2. the method according to claim 1 for extracting microbial oil, which is characterized in that in step (2), pH 3.5.
3. the method according to claim 1 for extracting microbial oil, which is characterized in that in step (2), isopropanol ethyl alcohol
The volume ratio of isopropanol and ethyl alcohol is 1:(7~10 in mixed solution).
4. the method according to claim 3 for extracting microbial oil, which is characterized in that in step (2), isopropanol ethyl alcohol
The volume ratio of isopropanol and ethyl alcohol is 1:8 in mixed solution.
5. the method according to claim 1 for extracting microbial oil, which is characterized in that wet thallus and second in step (1)
The solid-liquid ratio g/mL of alcohol solution is 1:(3~5).
6. the method according to claim 5 for extracting microbial oil, which is characterized in that wet thallus and second in step (1)
The solid-liquid ratio g/mL of alcohol solution is 1:3.
7. the method according to claim 1 for extracting microbial oil, which is characterized in that the water content of wet thallus is
50%.
8. the method according to claim 1 for extracting microbial oil, which is characterized in that step (3) are as follows: tetrahydro furan is added
It mutters, adjusts pH 2.2, separation of solid and liquid obtains oil-containing mixed solvent after 1h is extracted in -5 DEG C of concussions；Wherein, the additional amount of tetrahydrofuran
It is 2 times of feed liquid total volume obtained by step (2).
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| CN201811157869.1A CN109022141A (en) | 2018-09-30 | 2018-09-30 | A method of extracting microbial oil |
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Citations (5)
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| CN102724887A (en) * | 2010-01-29 | 2012-10-10 | 科学发展实验室 | Solid/liquid extraction |
| CN103789083A (en) * | 2014-02-17 | 2014-05-14 | 南京工业大学大丰海洋产业研究院 | Method for extracting fungal oil |
| CN103937604A (en) * | 2014-04-08 | 2014-07-23 | 北京化工大学 | Method for extracting oil in microalgae |
-
2018
- 2018-09-30 CN CN201811157869.1A patent/CN109022141A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010138620A1 (en) * | 2009-05-26 | 2010-12-02 | Solazyme, Inc. | Fractionation of oil-bearing microbial biomass |
| CN102724958A (en) * | 2010-01-29 | 2012-10-10 | 科学发展实验室 | Liquid/liquid extraction |
| CN102724887A (en) * | 2010-01-29 | 2012-10-10 | 科学发展实验室 | Solid/liquid extraction |
| CN103789083A (en) * | 2014-02-17 | 2014-05-14 | 南京工业大学大丰海洋产业研究院 | Method for extracting fungal oil |
| CN103937604A (en) * | 2014-04-08 | 2014-07-23 | 北京化工大学 | Method for extracting oil in microalgae |
Non-Patent Citations (2)
| Title |
|---|
| RUI HUANG, JUN CHENG, YI QIU, TAO LI, JUNHU ZHOU, KEFA CEN: "Using renewable ethanol and isopropanol for lipid transesterification in wet microalgae cells to produce biodiesel with low crystallization temperature", 《ENERGY CONVERSION AND MANAGEMENT》 * |
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