CN101307341A - Production process of docosahexenoic acid grease by bioenzyme method wall-breaking - Google Patents
Production process of docosahexenoic acid grease by bioenzyme method wall-breaking Download PDFInfo
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- CN101307341A CN101307341A CNA2008100478592A CN200810047859A CN101307341A CN 101307341 A CN101307341 A CN 101307341A CN A2008100478592 A CNA2008100478592 A CN A2008100478592A CN 200810047859 A CN200810047859 A CN 200810047859A CN 101307341 A CN101307341 A CN 101307341A
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- broken wall
- fermented liquid
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- DVSZKTAMJJTWFG-UHFFFAOYSA-N docosa-2,4,6,8,10,12-hexaenoic acid Chemical compound CCCCCCCCCC=CC=CC=CC=CC=CC=CC(O)=O DVSZKTAMJJTWFG-UHFFFAOYSA-N 0.000 title claims abstract description 49
- 239000004519 grease Substances 0.000 title claims abstract description 34
- MBMBGCFOFBJSGT-KUBAVDMBSA-N docosahexaenoic acid Natural products CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCC(O)=O MBMBGCFOFBJSGT-KUBAVDMBSA-N 0.000 title claims description 94
- 238000004519 manufacturing process Methods 0.000 title claims description 23
- 238000000034 method Methods 0.000 title abstract description 19
- 239000007788 liquid Substances 0.000 claims abstract description 59
- 239000010779 crude oil Substances 0.000 claims abstract description 32
- 241000199912 Crypthecodinium cohnii Species 0.000 claims abstract description 25
- 239000003921 oil Substances 0.000 claims abstract description 15
- 238000007670 refining Methods 0.000 claims abstract description 15
- 102000004190 Enzymes Human genes 0.000 claims description 53
- 108090000790 Enzymes Proteins 0.000 claims description 53
- 229940088598 enzyme Drugs 0.000 claims description 53
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 26
- 229940090949 docosahexaenoic acid Drugs 0.000 claims description 23
- 235000020669 docosahexaenoic acid Nutrition 0.000 claims description 23
- 239000000284 extract Substances 0.000 claims description 13
- 108010059892 Cellulase Proteins 0.000 claims description 11
- 238000009874 alkali refining Methods 0.000 claims description 11
- 229940106157 cellulase Drugs 0.000 claims description 11
- 238000004332 deodorization Methods 0.000 claims description 11
- 108010001682 Dextranase Proteins 0.000 claims description 9
- 108090000145 Bacillolysin Proteins 0.000 claims description 7
- 102000035092 Neutral proteases Human genes 0.000 claims description 7
- 108091005507 Neutral proteases Proteins 0.000 claims description 7
- 238000000855 fermentation Methods 0.000 claims description 6
- 230000004151 fermentation Effects 0.000 claims description 6
- 239000003960 organic solvent Substances 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 3
- 210000004027 cell Anatomy 0.000 abstract description 4
- 210000002421 cell wall Anatomy 0.000 abstract description 4
- 238000001035 drying Methods 0.000 abstract description 3
- 150000002978 peroxides Chemical class 0.000 abstract description 3
- 230000008901 benefit Effects 0.000 abstract description 2
- 230000003647 oxidation Effects 0.000 abstract description 2
- 238000007254 oxidation reaction Methods 0.000 abstract description 2
- 238000012258 culturing Methods 0.000 description 9
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 8
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical group CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 7
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 238000000638 solvent extraction Methods 0.000 description 3
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000012262 fermentative production Methods 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 208000024827 Alzheimer disease Diseases 0.000 description 1
- 108090000317 Chymotrypsin Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 244000227633 Ocotea pretiosa Species 0.000 description 1
- 235000004263 Ocotea pretiosa Nutrition 0.000 description 1
- 206010039966 Senile dementia Diseases 0.000 description 1
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 1
- 239000004141 Sodium laurylsulphate Substances 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000013043 chemical agent Substances 0.000 description 1
- 238000001311 chemical methods and process Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 229960002376 chymotrypsin Drugs 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000000366 juvenile effect Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000007514 neuronal growth Effects 0.000 description 1
- 238000000053 physical method Methods 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 235000019333 sodium laurylsulphate Nutrition 0.000 description 1
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Fats And Perfumes (AREA)
Abstract
The invention relates to a method for making docosahexaenic acid (DHA) grease through adopting bio-enzyme method wall breaking, comprising the following steps that: Crypthecodinium cohnii(Seligo)Javornicky is taken as starting strain; alga cell is obtained through carrying out continuous cultivation of Crypthecodinium cohnii(Seligo)Javornicky in liquid medium; and then DHA grease is extracted from the alga cell through bio-enzyme method wall breaking. The method has the advantages that: alga cell wall breaking is complete with the wall-breaking rate reaching more than 95 percent; the method does not need a drying step, and can prevent the oxidation of DHA crude oil, thereby ensuring that the peroxide value of extracted grease is extremely low (i.e. the POV value is less than 0.5); moreover, the yield of the extracted grease is more than 90 percent of total oil, while the refining yield is more than 70 percent of crude oil.
Description
Technical field
The present invention relates to utilize Kou Shi Crypthecodinium cohnii [Crypthecodinium cohnii (Seligo) Javornicky] to produce docosahexenoic acid (DHA) grease technical field, specifically a kind of biological enzyme broken wall is used for the docosahexaenoic acid grease production method.
Background technology
In recent years, (Polyunsaturated fatty acids, PUFAs) vital role to human health more and more is subject to people's attention polyunsaturated fatty acid.Wherein, docosahexenoic acid because of its in human body and extremely people's the favor of the intravital important physiological function of animal.Studies show that DHA is the basal component of cytolemma in some tissue of human body; Infant's vision system and neural growth are played an important role; Also have the reducing cholesterol effect, anticoagulant efficacy and inhibition cancer effect prevent senile dementia etc., and it are the necessary a kind of lipid acid of juvenile growth.Oneself forms suitability for industrialized production to utilize the Kou Shi Crypthecodinium cohnii to produce docosahexenoic acid (DHA) grease, and the production technique of docosahexenoic acid is being updated.Report openly now that wherein the frustule wall-breaking method has two classes in marine microalgae production docosahexenoic acid (DHA) grease.One class is the physical method broken wall: mechanical breaking-wall method grinds sassafras as machineries such as employing clarifixator or colloidal mills and extruding makes cell wall rupture, desiccating method etc.Adopting mechanical breaking-wall method method cell wall breaking incomplete, is total oily 60-70% with the organic solvent extraction recovery rate.Adopt the desiccating method time consumption and energy consumption, and it is higher to extract the grease peroxide value after heating and the vent method drying, the crude oil inferior quality, the lyophilize cost is higher, dry back with the organic solvent extraction refining after yield only be crude oil 30-50%.Open day is that disclosed to utilize marine microalgae to produce the greasy method of docosahexenoic acid (DHA) be to extract after adopting lyophilize or heating and vent method drying for the patent application of CN1101034A for April 5 nineteen ninety-five, publication number.Another kind of is the chemical process broken wall; Adopt chemical reagent to destroy cell wall structure, obtain crude oil with organic solvent extraction then.Open day is on June 27th, 2007, publication number is that the disclosed method with Kou Shi Crypthecodinium cohnii fermentative production docosahexaenoic acid grease of the patent application of CN1986822A is to adopt to add broken wall agent such as sodium lauryl sulphate etc., and its consumption is the 0.1-0.6% of concentrated broth weight.This method obtains having in the grease chemical agent residue, the quality of influence oil.And quality of crude oil is relatively poor, and yield only is crude oil 30-50% after the refining.
Summary of the invention
The objective of the invention is to develop and a kind of enzymatic shell-broken is used for the method for Kou Shi Crypthecodinium cohnii [Crypthecodinium cohnii (Seligo) Javornicky] fermentative production docosahexaenoic acid grease, it has improved the recovery rate of docosahexenoic acid (being called for short DHA) and the quality of crude oil widely.
Biological enzyme broken wall of the present invention is used for the docosahexaenoic acid grease production method, comprise: [Crypthecodinium cohnii (Seligo) Javornicky] is starting strain with the Kou Shi Crypthecodinium cohnii, cultured continuously Kou Shi Crypthecodinium cohnii obtains alginic cell in the liquid medium within, extracts docosahexaenoic acid grease by the biological enzyme broken wall from alginic cell then.
The used Kou Shi Crypthecodinium cohnii strain name of the present invention is Crypthecodinium cohnii (Seligo) Javornicky.Can cultivate mechanism from the preservation that comprises ATCC (U.S. typical case's culture collection center) obtains.
Biological enzyme is divided into main body enzyme and auxiliary enzymes;
The main body enzyme is Sumizyme MP (2709), neutral protease, papoid, can use wherein one or more, if several, its ratio can be used by arbitrary proportion.Its consumption is the 0.12-0.5% of fermented liquid weight, if concentrated broth, its consumption calculates by concentrating primary fermentation liquid weight;
Auxiliary enzymes is trypsinase, Alphachymdean Catarasce, and cellulase, dextranase, zytase can use wherein one or more, if multiple, its ratio can be used by arbitrary proportion.Its consumption is the 0-0.025% of fermented liquid weight, if concentrated broth, enzyme dosage calculates by concentrating primary fermentation liquid weight;
In use, separately with main body enzyme or main body enzyme and auxiliary enzymes and with all reaching broken wall.
Biological enzyme broken wall of the present invention is used for the docosahexaenoic acid grease production method: add the main body enzyme at fermented liquid or filtering and concentrating fermented liquid, if consumption is the 0.12-0.5% concentrated broth of fermented liquid weight, its consumption calculates by concentrating primary fermentation liquid weight; Auxiliary enzymes, consumption is the 0-0.025% of fermented liquid weight, if concentrated broth, enzyme dosage calculates by concentrating primary fermentation liquid weight, and intensification 50-70 ℃, insulated and stirred 3-9 hour, the ethanol of adding 95%, the ethanol consumption is the 30%-150% of fermented liquid concentrated broth, and adding organic solvent amount is fermented liquid or concentrated broth weight 1.5-4.5 a times, extracts and obtains the docosahexenoic acid crude oil.The docosahexenoic acid crude oil is obtained the docosahexenoic acid refining oil after aquation, alkali refining, decolouring, deodorization.
Wherein said organic solvent is normal hexane or chloroform etc.
The add-on of main body enzyme and auxiliary enzymes in the concentrated broth of the present invention is to calculate with the fermentation liquid measure before the filtering and concentrating fermented liquid.
The advantage that biological enzyme broken wall of the present invention is used for the docosahexaenoic acid grease production method is: the frustule broken wall is complete, sporoderm-broken rate is more than 95%, do not need through super-dry, can prevent the oxidation of docosahexenoic acid crude oil, make the oil and grease extracting peroxide value extremely low, the POV value is below 0.5, and recovery rate is more than 90% of total oil, and refining yield is more than 70% of crude oil.
Embodiment
Embodiment one:
A kind of biological enzyme broken wall is used for the docosahexaenoic acid grease production method: adopt Kou Shi Crypthecodinium cohnii slant strains to carry out liquid shaking bottle cultivation, liquid shaking bottle enlarged culturing, first order seed cultivation, secondary seed cultivation and three grade fermemtation and cultivate, obtain fermented liquid.Add Sumizyme MP (2709) 0.2kg in fermented liquid 100kg, intensification 50-70 ℃, insulated and stirred 3-9 hour, the ethanol 30kg of adding 95% added normal hexane 150kg, extracts and obtains docosahexenoic acid crude oil 3kg.The docosahexenoic acid crude oil is obtained docosahexenoic acid refining oil 2.34kg after aquation, alkali refining, decolouring, deodorization.
Embodiment two:
A kind of biological enzyme broken wall is used for the docosahexaenoic acid grease production method: adopt Kou Shi Crypthecodinium cohnii slant strains to carry out liquid shaking bottle cultivation, liquid shaking bottle enlarged culturing, first order seed cultivation, secondary seed cultivation and three grade fermemtation and cultivate, obtain fermented liquid.Add neutral protease 0.35kg in fermented liquid 100kg, intensification 50-70 ℃, insulated and stirred 3-9 hour, the ethanol 30kg of adding 95% added normal hexane 150kg, extracts and obtains docosahexenoic acid crude oil 3kg.The docosahexenoic acid crude oil is obtained docosahexenoic acid refining oil 2.34kg after aquation, alkali refining, decolouring, deodorization.
Embodiment three:
A kind of biological enzyme broken wall is used for the docosahexaenoic acid grease production method: adopt Kou Shi Crypthecodinium cohnii slant strains to carry out liquid shaking bottle cultivation, liquid shaking bottle enlarged culturing, first order seed cultivation, secondary seed cultivation and three grade fermemtation and cultivate, obtain fermented liquid.Add papoid 0.45kg in fermented liquid 100kg, intensification 50-70 ℃, insulated and stirred 3-9 hour, the ethanol 30kg of adding 95% added normal hexane 150kg, extracts and obtains docosahexenoic acid crude oil 3kg.The docosahexenoic acid crude oil is obtained docosahexenoic acid refining oil 2.34kg after aquation, alkali refining, decolouring, deodorization.
Embodiment four:
A kind of biological enzyme broken wall is used for the docosahexaenoic acid grease production method: adopt Kou Shi Crypthecodinium cohnii slant strains to carry out liquid shaking bottle cultivation, liquid shaking bottle enlarged culturing, first order seed cultivation, secondary seed cultivation and three grade fermemtation and cultivate, obtain fermented liquid.In fermented liquid 100kg, add Sumizyme MP (2709) 0.12kg, add Chymotrypsin 0.001kg, intensification 50-70 ℃, insulated and stirred 3-9 hour, the ethanol 30kg of adding 95% adds normal hexane 150kg, extracts and obtains docosahexenoic acid crude oil 3kg.The docosahexenoic acid crude oil is obtained docosahexenoic acid refining oil 2.34kg after aquation, alkali refining, decolouring, deodorization.
Embodiment five:
A kind of biological enzyme broken wall is used for the docosahexaenoic acid grease production method: adopt Kou Shi Crypthecodinium cohnii slant strains to carry out liquid shaking bottle cultivation, liquid shaking bottle enlarged culturing, first order seed cultivation, secondary seed cultivation and three grade fermemtation and cultivate, obtain fermented liquid.In fermented liquid 100kg, add neutral protease 0.18kg, add trypsinase 0.05kg, Alphachymdean Catarasce 0.01kg altogether, intensification 50-70 ℃, insulated and stirred 3-9 hour, the ethanol 60kg of adding 95% adds chloroform 200kg, extracts and obtains docosahexenoic acid crude oil 3.8kg.The docosahexenoic acid crude oil is obtained docosahexenoic acid refining oil 3.04kg after aquation, alkali refining, decolouring, deodorization.
Embodiment six:
A kind of biological enzyme broken wall is used for the docosahexaenoic acid grease production method: adopt Kou Shi Crypthecodinium cohnii slant strains to carry out liquid shaking bottle and cultivate, the liquid shaking bottle enlarged culturing, first order seed is cultivated, secondary seed is cultivated and three grade fermemtation is cultivated, obtain fermented liquid, in centrifugal concentrated broth 100kg (being 400kg before fermented liquid is centrifugal), add Sumizyme MP (2709) 0.8kg, in cellulase: dextranase: zytase is that 1: 3: 1 ratio adds cellulase, dextranase, zytase is 0.015kg altogether, intensification 50-70 ℃, insulated and stirred 3-9 hour, the ethanol 90kg of adding 95%, add normal hexane 250kg, extract and obtain docosahexenoic acid crude oil 16.0kg.The docosahexenoic acid crude oil is obtained docosahexenoic acid refining oil 12kg after aquation, alkali refining, decolouring, deodorization.
Embodiment seven:
A kind of biological enzyme broken wall is used for the docosahexaenoic acid grease production method: adopt Kou Shi Crypthecodinium cohnii slant strains to carry out liquid shaking bottle and cultivate, the liquid shaking bottle enlarged culturing, first order seed is cultivated, secondary seed is cultivated and three grade fermemtation is cultivated, obtain fermented liquid, in centrifugal concentrated broth 100kg (being 400kg before fermented liquid is centrifugal), add neutral protease and papoid 0.6kg altogether equably, in trypsinase: Alphachymdean Catarasce: cellulase is that 2: 3: 5 ratio adds trypsinase, Alphachymdean Catarasce, cellulase is 0.018kg altogether, intensification 50-70 ℃, insulated and stirred 3-9 hour, the ethanol 120kg of adding 95%, add chloroform 300kg, extract and obtain docosahexenoic acid crude oil 12.8kg.The docosahexenoic acid crude oil is obtained docosahexenoic acid refining oil 9.2kg after aquation, alkali refining, decolouring, deodorization.
Embodiment eight:
A kind of biological enzyme broken wall is used for the docosahexaenoic acid grease production method: adopt Kou Shi Crypthecodinium cohnii slant strains to carry out liquid shaking bottle and cultivate, the liquid shaking bottle enlarged culturing, first order seed is cultivated, secondary seed is cultivated and three grade fermemtation is cultivated, obtain fermented liquid, in centrifugal concentrated broth 100kg (being 450kg before fermented liquid is centrifugal), add Sumizyme MP (2709) and papoid 0.9kg altogether equably, in trypsinase: Alphachymdean Catarasce: cellulase: dextranase is that 1: 4: 1 ratio adds trypsinase, Alphachymdean Catarasce, cellulase, dextranase is 0.02kg altogether, intensification 50-70 ℃, insulated and stirred 3-9 hour, the ethanol 135kg of adding 95%, add normal hexane 350kg, extract and obtain docosahexenoic acid crude oil 14kg.The docosahexenoic acid crude oil is obtained docosahexenoic acid refining oil 10kg after aquation, alkali refining, decolouring, deodorization.
Embodiment nine:
A kind of biological enzyme broken wall is used for the docosahexaenoic acid grease production method: adopt Kou Shi Crypthecodinium cohnii slant strains to carry out liquid shaking bottle cultivation, liquid shaking bottle enlarged culturing, first order seed cultivation, secondary seed cultivation and three grade fermemtation and cultivate, obtain fermented liquid.In fermented liquid 100kg, add neutral protease 0.5kg, in trypsinase: Alphachymdean Catarasce: cellulase: dextranase: zytase is 1: 2: 1: 1: 1 ratio adds trypsinase, Alphachymdean Catarasce, cellulase, dextranase, zytase be 0.025kg altogether, intensification 50-70 ℃, insulated and stirred 3-9 hour, the ethanol 150kg of adding 95% adds chloroform 450kg, extracts and obtains docosahexenoic acid crude oil 3.5kg.The docosahexenoic acid crude oil is obtained docosahexenoic acid refining oil 2.6kg after aquation, alkali refining, decolouring, deodorization.
Claims (5)
1, a kind of biological enzyme broken wall is used for the docosahexaenoic acid grease production method, it is characterized in that: comprising: [Crypthecodinium cohnii (Seligo) Javornicky] is starting strain with the Kou Shi Crypthecodinium cohnii, cultured continuously Kou Shi Crypthecodinium cohnii obtains alginic cell in the liquid medium within, extracts docosahexaenoic acid grease by the biological enzyme broken wall from alginic cell then.
2, biological enzyme broken wall according to claim 1 is used for the docosahexaenoic acid grease production method, it is characterized in that: in fermented liquid or filtering and concentrating fermented liquid, add main body enzyme, auxiliary enzymes, intensification 50-70 ℃, insulated and stirred 3-9 hour, the ethanol of adding 95%, the ethanol consumption is the 30%-150% of fermented liquid, and the adding organic solvent extracts and obtains the docosahexenoic acid crude oil; The docosahexenoic acid crude oil is obtained the docosahexenoic acid refining oil after aquation, alkali refining, decolouring, deodorization.
3, biological enzyme broken wall according to claim 1 is used for the docosahexaenoic acid grease production method, it is characterized in that: biological enzyme is divided into main body enzyme and auxiliary enzymes; The main body enzyme is Sumizyme MP (2709), neutral protease, papoid; Auxiliary enzymes is trypsinase, Alphachymdean Catarasce, cellulase, dextranase, zytase; In use, separately with main body enzyme or main body enzyme and auxiliary enzymes and with all reaching broken wall.
4, biological enzyme broken wall according to claim 3 is used for the docosahexaenoic acid grease production method, it is characterized in that: biological enzyme is divided into main body enzyme and auxiliary enzymes; The main body enzyme dosage is the 0.12-0.5% of fermented liquid weight; The auxiliary enzymes consumption is the 0-0.025% of fermented liquid weight, if concentrated broth, enzyme dosage calculates by concentrating primary fermentation liquid weight.
5, biological enzyme broken wall according to claim 3 is used for the docosahexaenoic acid grease production method, it is characterized in that: the main body enzyme is Sumizyme MP (2709), neutral protease, papoid, should be wherein one or more during use, if several, its ratio is used by arbitrary proportion; Auxiliary enzymes is trypsinase, Alphachymdean Catarasce, cellulase, dextranase, zytase; Should be wherein one or more during use, if multiple, its ratio is used by arbitrary proportion.
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Cited By (14)
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| CN101824363A (en) * | 2010-05-25 | 2010-09-08 | 南京工业大学 | Method for extracting docosahexaenoic acid grease |
| CN101985637A (en) * | 2010-11-02 | 2011-03-16 | 嘉吉烯王生物工程（武汉）有限公司 | Method for extracting microbial oil |
| CN102199482A (en) * | 2011-04-15 | 2011-09-28 | 北京化工大学 | Method for extracting grease from oleaginous microorganisms |
| CN102199485A (en) * | 2011-04-29 | 2011-09-28 | 国家海洋局第三海洋研究所 | Method for extracting schizochytrium aggregatum oil |
| CN101585759B (en) * | 2009-07-08 | 2012-05-23 | 内蒙古金达威药业有限公司 | Method of extracting DHA unsaturated fatty acid from dino flagellate fermentation liquor |
| CN102492544A (en) * | 2011-12-07 | 2012-06-13 | 湖北福星生物科技有限公司 | Method for preparing microalgal docosahexaenoic acid (DHA) oil by dry method |
| CN102533879A (en) * | 2010-12-17 | 2012-07-04 | 中国科学院大连化学物理研究所 | Microbial oil extraction method |
| CN102887821A (en) * | 2012-09-29 | 2013-01-23 | 青岛琅琊台集团股份有限公司 | Method for extracting DHA (docosahexaenoic acid) through extraction and separation of marine microalgae fermentation liquid |
| CN102976936A (en) * | 2012-11-16 | 2013-03-20 | 成都圆大生物科技有限公司 | Method of reducing peroxide value and anisidine value in unsaturated fatty acid ethyl ester |
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| WO2006046943A2 (en) * | 2004-10-22 | 2006-05-04 | Martek Biosciences Corporation | Methods for producing lipids by liberation from biomass |
| CN1986822A (en) * | 2006-12-15 | 2007-06-27 | 湖北友芝友生物科技有限公司 | Crypthecodinium connii fermenting process for producing docosahexaenoic acid grease |
| CN101168501B (en) * | 2007-07-11 | 2010-04-07 | 南京工业大学 | A process for extracting and refining fatty acid rich in DHA from Cryptidinium |
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- 2008-05-29 CN CN2008100478592A patent/CN101307341B/en not_active Expired - Fee Related
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| CN101585759B (en) * | 2009-07-08 | 2012-05-23 | 内蒙古金达威药业有限公司 | Method of extracting DHA unsaturated fatty acid from dino flagellate fermentation liquor |
| CN101824363A (en) * | 2010-05-25 | 2010-09-08 | 南京工业大学 | Method for extracting docosahexaenoic acid grease |
| CN101824363B (en) * | 2010-05-25 | 2012-04-25 | 南京工业大学 | Method for extracting docosahexaenoic acid grease |
| CN101985637B (en) * | 2010-11-02 | 2014-05-07 | 嘉必优生物工程（武汉）有限公司 | Method for extracting microbial oil |
| CN101985637A (en) * | 2010-11-02 | 2011-03-16 | 嘉吉烯王生物工程（武汉）有限公司 | Method for extracting microbial oil |
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| CN102533879B (en) * | 2010-12-17 | 2013-12-04 | 中国科学院大连化学物理研究所 | Microbial oil extraction method |
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| CN109181842A (en) * | 2018-08-20 | 2019-01-11 | 梁云 | Microbial oil and its extracting method |
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