CN104962589B - A kind of microbial oil and preparation method rich in phosphatide type polyunsaturated fatty acid - Google Patents
A kind of microbial oil and preparation method rich in phosphatide type polyunsaturated fatty acid Download PDFInfo
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- CN104962589B CN104962589B CN201510473476.1A CN201510473476A CN104962589B CN 104962589 B CN104962589 B CN 104962589B CN 201510473476 A CN201510473476 A CN 201510473476A CN 104962589 B CN104962589 B CN 104962589B
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- microbial oil
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- phosphatide
- obtains
- fatty acid
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- 230000000813 microbial effect Effects 0.000 title claims abstract description 82
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 title claims abstract description 44
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- 239000004519 grease Substances 0.000 claims abstract description 15
- 238000000605 extraction Methods 0.000 claims abstract description 13
- MBMBGCFOFBJSGT-KUBAVDMBSA-N all-cis-docosa-4,7,10,13,16,19-hexaenoic acid Chemical compound 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 claims description 44
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 38
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- 235000020669 docosahexaenoic acid Nutrition 0.000 claims description 22
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- 150000003904 phospholipids Chemical class 0.000 claims description 21
- JAZBEHYOTPTENJ-JLNKQSITSA-N all-cis-5,8,11,14,17-icosapentaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O JAZBEHYOTPTENJ-JLNKQSITSA-N 0.000 claims description 13
- 235000020673 eicosapentaenoic acid Nutrition 0.000 claims description 13
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- JAZBEHYOTPTENJ-UHFFFAOYSA-N eicosapentaenoic acid Natural products CCC=CCC=CCC=CCC=CCC=CCCCC(O)=O JAZBEHYOTPTENJ-UHFFFAOYSA-N 0.000 claims description 13
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- DTOSIQBPPRVQHS-PDBXOOCHSA-N alpha-linolenic acid Chemical compound CC\C=C/C\C=C/C\C=C/CCCCCCCC(O)=O DTOSIQBPPRVQHS-PDBXOOCHSA-N 0.000 claims description 6
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- KQQKGWQCNNTQJW-UHFFFAOYSA-N linolenic acid Natural products CC=CCCC=CCC=CCCCCCCCC(O)=O KQQKGWQCNNTQJW-UHFFFAOYSA-N 0.000 claims description 6
- 239000010409 thin film Substances 0.000 claims description 6
- 241000003595 Aurantiochytrium limacinum Species 0.000 claims description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 4
- 108091005804 Peptidases Proteins 0.000 claims description 4
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- YZXBAPSDXZZRGB-DOFZRALJSA-N arachidonic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O YZXBAPSDXZZRGB-DOFZRALJSA-N 0.000 description 12
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 description 10
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- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- 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
- C11B1/02—Pretreatment
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6436—Fatty acid esters
- C12P7/6445—Glycerides
- C12P7/6463—Glycerides obtained from glyceride producing microorganisms, e.g. single cell oil
-
- 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
- C11B1/02—Pretreatment
- C11B1/025—Pretreatment by enzymes or microorganisms, living or dead
-
- 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
- C11B1/106—Production of fats or fatty oils from raw materials by extracting using ultra-sounds
-
- 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
-
- 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
- C11B3/00—Refining fats or fatty oils
- C11B3/001—Refining fats or fatty oils by a combination of two or more of the means hereafter
-
- 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
- C11B3/00—Refining fats or fatty oils
- C11B3/006—Refining fats or fatty oils by extraction
-
- 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
- C11B3/00—Refining fats or fatty oils
- C11B3/008—Refining fats or fatty oils by filtration, e.g. including ultra filtration, dialysis
-
- 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
- C11B3/00—Refining fats or fatty oils
- C11B3/02—Refining fats or fatty oils by chemical reaction
-
- 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
- C11B3/00—Refining fats or fatty oils
- C11B3/10—Refining fats or fatty oils by adsorption
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6436—Fatty acid esters
- C12P7/6445—Glycerides
- C12P7/6472—Glycerides containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone
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Abstract
The present invention provides a kind of preparation method of microbial oil rich in phosphatide type polyunsaturated fatty acid, include the following steps: that (1) fermentation obtains the microorganism for being rich in polyunsaturated fatty acid, (2) extraction is carried out to microorganism, obtain microbial oil, (3) phosphatide in enriched microorganism grease obtains the microbial oil rich in phosphatide type polyunsaturated fatty acid after purification.The invention has the following advantages: 1, entire preparation method is using natural microbial oil as raw material, therefore raw material is unrestricted, 2, content of polyunsaturated fatty acid is high in the phosphatide type polyunsaturated fatty acid microbial oil prepared, 3, compared with the phosphatide type polyunsaturated fatty acid of existing enzymatic clarification, safer, application range is more extensive.
Description
Technical field
The microbial oil and preparation method thereof rich in phosphatide type polyunsaturated fatty acid that the present invention relates to a kind of.
Background technique
Phosphatide is the lipids containing phosphoric acid, is prevalent in biological cell matter and cell membrane, thin to maintenance
After birth function maintains cell metabolism to play a crucial role in turn.Phosphatide can be divided into glyceric alcohol phosphatides and sphingomyelins two by its molecular structure
Major class.Glyceric alcohol phosphatides mainly include lecithin, cephalin, serinephosphatide, lipositol etc..It is low for nervous system function
Lower bring disease such as lather, loss of appetite, tinnitus, impotence power, nerve diarrhea, practises perseverance constipation and nerve
Dementia brought by material want and the decline of memory power are conveyed, lecithin is fully supplemented daily, will receive good curative effect.
Studies have found that in intelligence test (labyrinth test), memory is significantly better than offspring with the rat of lecithin feeding pregnancy
The offspring of the rat of lecithin is not raised.7th lecithin international conference in 1997 was once drawn a conclusion: " it is recommended that pregnant woman takes
With suitable lecithin, this is critically important for the intellectual development of child." 1998 years lecithin are by FAO (Food and Agriculture Organization of the United Nation)
(FAO) it is classified as one of big nutraceutical in the world five;U.S.'s food in 1999 and the medication management committee (FDA) are clearly stipulate that institute
Lecithin will be supplemented in right amount in some baby's recipes.
Phosphatide is mainly derived from plant at present, and commercially available phosphatide mainly contains palmitinic acid, oleic acid, linoleic acid and stearic acid etc.
Saturated fatty acid or monounsaturated fatty acids containing 16 or 18 carbon atoms.However other than these fatty acid, human body is also needed
Polyunsaturated fatty acid is supplemented, such as linolenic acid, arachidonic acid, eicosapentaenoic acid, clupanodonic acid and 22
Carbon acid etc..Docosahexaenoic acid in the brain dense is considered in vision, cognitive function and other is normal
Vital effect is played in the development of brain function.Arachidonic acid is most abundant and most active in the mammalian body
A kind of polyunsaturated fatty acid, it not only has particularly important meaning to the health of human body, also extremely to the normal development of infant
Close it is important, arachidonic acid also have prevention cardiovascular disease, immunological regulation, auxiliary inhibit tumour, prevention canceration and adjust mind
The function of warp can prevent and treat the diseases such as hypertension, diabetes, obesity and virus infection.
Compared with methyl esters/ethyl ester type or triglyceride type polyunsaturated fatty acid, phosphatide type polyunsaturated fatty acid is not only
, the advantages such as highly-safe and stability good high with bioavilability, moreover it is possible to while phosphatide abundant is provided for body, to brain
Development play the effects of 1+1 > 2.Some researches show that human body only has the digestibility of ethyl ester type docosahexaenoic acid
21%, and be more than 99% to the digestibility of phosphatide type docosahexaenoic acid.Docosahexaenoic acid and yolk in yolk
In lecithin be closely linked, be digested together with lecithin, be particularly suitable for pregnant and lying-in women and infant food
With.
China Patent Publication No. is that the patent of invention of CN103509047A discloses a kind of phosphatidyl of antarctic krill
The extraction process of choline and the preparation method of phosphatidylserine.This method is by collecting krill head and by decocting stirring
It is 7.6-8.1 %, Content of Eicosapentaenoic Acid 14.6- that the series of process such as extraction, which obtain docosahexaenoic acid content,
15.6% phospholipid prod.The drawbacks of this method is: raw material sources have larger limitation, and the processes such as decoction can be lost largely
Polyunsaturated fatty acid, and the fishing without restraint of krill can damage environment.
China Patent Publication No. is that the patent of invention of CN101195637B discloses a kind of prepare rich in polyunsaturated fat
The technique of sour phosphatide, this method mainly pass through enzyme to soybean lecithin and rich in eicosapentaenoic acid and docosahexaenoic acid
Free fatty acid carries out transesterification catalysis reaction, to obtain the phosphatide for being rich in polyunsaturated fatty acid.But this method system
In the phosphatide obtained, eicosapentaenoic acid and docosahexaenoic acid content are lower.And this method is needed using enzyme preparation, because
The application range of this finished product will receive limitation.
To sum up, a kind of useful environment is provided, sustainable development, with wider application range rich in phosphatide type mostly not
Microbial oil of saturated fatty acid and preparation method thereof is actually necessary.
Summary of the invention
The object of the present invention is to provide a kind of production requirement for being able to satisfy industrialization, be rich in phosphatide type polyunsaturated fat
The preparation method of the microbial oil of acid.
It is a further object of the present invention to provide one kind to have high-content phosphatide, and has high-content how unsaturated rouge in phosphatide
The microbial oil of fat acid.
To realize that above-mentioned main purpose, the present invention provide a kind of microbial oil rich in phosphatide type polyunsaturated fatty acid
Preparation method, include the following steps: (1) fermentation obtain be rich in polyunsaturated fatty acid microorganism, (2) to microorganism carry out
Extraction, obtains microbial oil, and the phosphatide in (3) enriched microorganism grease is obtained after purification rich in the more insatiable hungers of phosphatide type
With the microbial oil of fatty acid.
The preparation method of microbial oil rich in phosphatide type polyunsaturated fatty acid of the invention has below beneficial to effect
Fruit: 1, entire preparation method is using natural microbial oil as raw material, therefore raw material is unrestricted, 2, the phosphatide type prepared
Content of polyunsaturated fatty acid is high in polyunsaturated fatty acid microbial oil, 3, the more insatiable hungers of phosphatide type with existing enzymatic clarification
It is compared with fatty acid, safer, application range is more extensive.
Further, real with stand at low temperature, solvent elution, supercritical extract, membrane separation process, water or steam in step (3)
Apply enrichment.The means of these enrichments meet the relevant regulations of food safety.
Another object to realize the present invention, the present invention provide a kind of microorganism rich in phosphatide type polyunsaturated fatty acid
Grease, content of phospholipid is not less than 38% in the microbial oil, and the content of polyunsaturated fatty acid is not less than 30% in phosphatide.
Microbial oil rich in phosphatide type polyunsaturated fatty acid of the invention has the advantages that firstly, oil
Polyunsaturated fatty acid in rouge be it is microbe-derived, therefore, can adapt to large-scale production requirement, and edible more pacify
Entirely;Secondly, content of phospholipid in grease is high, and in phosphatide polyunsaturated fatty acid content it is also high, be more conducive to be absorbed, and seek
Form a point height.
Further, in the above-mentioned microbial oil rich in phosphatide type polyunsaturated fatty acid, how unsaturated rouge in phosphatide
Fat acid content is 30%-79%.
Further, in the above-mentioned microbial oil rich in phosphatide type polyunsaturated fatty acid, polyunsaturated fatty acid packet
Include linolenic acid, arachidonic acid, eicosapentaenoic acid, clupanodonic acid and docosahexaenoic acid.
Further, in the above-mentioned microbial oil rich in phosphatide type polyunsaturated fatty acid, polyunsaturated fatty acid is micro-
The source of bio-oil is Mortierella alpina, microballoon algae, yeast, schizochytrium limacinum, Dunaliella salina or dino flagellate.
Specific embodiment
Below with reference to specific example, the present invention is described in further detail.
Embodiment 1
Using schizochytrium limacinum as fermenting microbe, successively follow these steps to operate:
(1) fermentation obtains the microbial fermentation solution for being rich in docosahexaenoic acid;
(2) by above-mentioned fermentation liquid after UF membrane, the concentrated broth that water content is 68% is obtained, the 6L concentration is taken to ferment
Liquid, is added the alkali protease of 3ml, adjusts pH value to 8.0, is warming up to 45 DEG C of heat preservation 4h progress broken walls, then with 10L n-hexane into
Row extracts.Obtained extracting solution is filtered with 1um filter stick, and precipitation obtains total 511g microbial oil;
(3) 450g mentioned microorganism grease is taken, 450ml hexane is added, 0 DEG C of processing 2h is cooled to, then 0.2MPa's
Under environment, micro-filtration is kept the temperature with 0.2um pore size ceramic membrane, the semisolid that film retains is obtained into 11g rich in phosphorus through thin film evaporation
The microbial oil of epoxy-type docosahexaenoic acid.This step is the technique for carrying out enrichment and purification process to phosphatide simultaneously.Through
Detection, the content of phospholipid of the microbial oil are 69.7%, and in phosphatide polyunsaturated fatty acid total content (Eicosatetraenoic
Acid, eicosapentaenoic acid, docosahexaenoic acid, the sum of clupanodonic acid) it is 79%.
Embodiment 2
Using microballoon algae as fermenting microbe, successively follow these steps to operate:
(1) fermentation obtains the fermentation liquid of the microbial cells rich in eicosapentaenoic acid, and fermentation liquid is obtained after processing
Dry mycelium rich in eicosapentaenoic acid;
(2) it takes the above-mentioned dry mycelium of 1kg to carry out crushing broken wall with pulverizer, the ethyl alcohol that 1kg content is 95% is added and is used as folder
Band agent, uses supercritical CO2It is 40 DEG C in temperature, pressure extracts 2h under conditions of being 30Mpa, extraction time is 2 times.By what is obtained
Extracting solution is filtered with 1um filter stick, then at 40 DEG C, isolated 357g microbial oil under conditions of 25Mpa;
(3) 300g mentioned microorganism grease is taken, supercritical CO is used2Extraction, extracts under the conditions of temperature 50 C, pressure 35Mpa
5h is taken, so at 50 DEG C, the isolated 19g of the condition of 28MPa is rich in the arachidonic microbial oil of phosphatide type.This step is
The technique that enrichment and purification process are carried out to phosphatide simultaneously.Through detecting, content of phospholipid is 41.8% in the microbial oil, and phosphorus
The total content (the sum of arachidonic acid and eicosapentaenoic acid) of polyunsaturated fatty acid is 38.1% in rouge.
Embodiment 3
Using dino flagellate as fermenting microbe, successively follow these steps to operate:
(1) fermentation obtains the microbial fermentation solution for being rich in docosahexaenoic acid;
(2) above-mentioned fermentation liquid is centrifuged, obtains the concentrated broth that water content is 73%, takes the 50L concentrated broth, add
Enter 50ml alkali protease, adjusts pH value to 8.0, be warming up to 50 DEG C of heat preservation 3h and carry out broken wall, then mentioned with 100L n-hexane
It takes, extracting solution is filtered with 1um filter stick, and precipitation obtains total 4168g microbial oil;
(3) 3kg mentioned microorganism grease is taken, 9000ml n-hexane is added, 200g pure water is added, at 35-40 DEG C,
In the environment of 0.15MPa, with 0.2um pore size ceramic membrane micro-filtration, the semisolid that film is retained obtains 230g through thin film evaporation
Microbial oil rich in phosphatide type docosahexaenoic acid.This step is to carry out the work of enrichment and purification process to phosphatide simultaneously
Skill.Through detecting, the content of phospholipid of the microbial oil is 58%, and in phosphatide polyunsaturated fatty acid total content (22 carbon
The sum of acid and eicosapentaenoic acid) it is 43.1 %.
Embodiment 4
Using Mortierella alpina as fermenting microbe, successively follow these steps to operate:
(1) fermentation obtains the fermentation liquid for being rich in arachidonic microbial cells, after handling the fermentation liquid,
It obtains and contains arachidonic dry mycelium;
(2) 4kg dry mycelium is taken, 25L n-hexane is added, with colloid mill circulation shear extraction, extracting solution is filtered with 1um
Stick filtering, precipitation obtain total 2128g microbial oil;
(3) 4h is stood in the environment of taking 2000g mentioned microorganism grease to be placed in -10 DEG C, then normal temperature unfreezing, is poured out
Layer edible vegetable oil, obtains the thick concentrated phosphatide of 205g.This step is the process of enriched microorganism phospholipid in lipid;
(4) the thick concentrated phosphatide of above-mentioned 205g is warming up to 65 DEG C, the food-grade H that 4g concentration is 30% is added2O2Stirring is anti-
30min is answered, then heats to 85-90 DEG C, residual moisture and H are removed in the environment that vacuum degree is -0.095Mpa2O2.To moisture
After 0.5%, 50 DEG C are cooled to hereinafter, obtaining 193g rich in the arachidonic microbial oil of phosphatide type.It is detected, it should
The content of phospholipid of microbial oil is 38%, and in phosphatide polyunsaturated fatty acid total content (linolenic acid and arachidonic acid it
With) it is 59.1 %.
Embodiment 5
To produce linolenic saccharomycete as fermenting microbe, successively follow these steps to operate:
(1) fermentation obtains the fermentation liquid for being rich in linolenic microbial cells, and fermentation liquid is obtained after processing rich in Asia
The dry mycelium of numb acid;
(2) 9kg dry mycelium is taken, with Ultrasonic Pulverization, 90L n-hexane is added, extracting solution is filtered with 1um filter stick, and precipitation obtains
Total 3978g microorganism;
(3) take 3600g mentioned microorganism grease, be passed through vapor to 105 DEG C of processing 5min, be then cooled to 80 DEG C from
Gains in depth of comprehension are to the thick concentrated phosphatide of 918g.This step is the phosphatide in enriched microorganism grease.
(4) by the thick concentrated phosphatide of 850g in 85-90 DEG C, vacuum degree -0.095Mpa removes the moisture in raw phospholipid.To thick
After moisture is less than 1% in phosphatide, it is cooled to 65 DEG C.Then 1800ml n-hexane, 25g active carbon, 50 DEG C of temperature control decolorations are added
30min.Obtained filtrate thin film evaporator precipitation is filtered, obtains 396g rich in the linolenic microbial oil of phosphatide type.Through
It detects, content of phospholipid is 63.7% in the microbial oil.And the content of polyunsaturated fatty acid (i.e. linolenic acid) is in phosphatide
30%。
Embodiment 6
Using Dunaliella salina as fermenting microbe, successively follow these steps to operate:
(1) fermentation obtains the fermentation liquid for being rich in linolenic microbial cells, after handling the fermentation liquid, obtains
Containing linolenic dry mycelium;
(2) 50kg dry mycelium is taken, 250L n-hexane is added, shears extraction with hand-held cutter, extracting solution is used
The filtering of 1um filter stick, precipitation obtain total 12.9kg microbial oil;
(3) 10kg mentioned microorganism grease is taken, is warming up to 80 DEG C, 1kg pure water is added and carries out hydration reaction 30min, then
Centrifugation obtains the thick concentrated phosphatide of 2.81kg.This step is the technique of the phosphatide in enriched microorganism grease.
(4) it by the thick concentrated phosphatide of 2.75kg in 85-90 DEG C, is removed in raw phospholipid in the environment of vacuum degree -0.095Mpa
Moisture.After moisture in raw phospholipid is less than 1%, it is cooled to 65-70 DEG C, the food-grade H that 10ml concentration is 30% is added2O2Stirring
30min is reacted, then heats to 85-90 DEG C, residual moisture and H are removed in the environment of vacuum degree -0.095Mpa2O2,To moisture
After 0.5%, 50 DEG C are cooled to hereinafter, obtaining 1.97kg rich in the linolenic microbial oil of phosphatide type.Through detecting, this is micro-
Content of phospholipid is 61.1% in bio-oil, and the total content (linolenic acid) of polyunsaturated fatty acid is 39.4% in phosphatide.
Embodiment 7
Using schizochytrium limacinum as fermenting microbe, successively follow these steps to operate:
(1) fermentation obtains the fermentation liquid of the microbial cells rich in docosahexaenoic acid, and fermentation liquid is obtained after processing
Obtain the dry mycelium rich in docosahexaenoic acid;
(2) it takes 500kg dry mycelium to carry out crushing broken wall with pulverizer, 2500L n-hexane is added and extracts, extracting solution 1um
Then filter stick filtering obtains total 247.3kg microbial oil through vacuum desolvation, the cold acetone removing grease of 800L, weight is added
It is 2 times multiple.Obtained wet solid is vacuum dried, removes acetone, obtains 7.7kg powder phospholipid, the powder phospholipid of the present embodiment is
A kind of specific form of microbial oil rich in phosphatide type polyunsaturated fatty acid.This step is purification step, main purpose
It is the impurity such as removal grease, solvent.Through detecting, content of phospholipid is 96.7% in the microbial oil, and how unsaturated rouge in phosphatide
The total content (eicosatetraenoic acid, eicosapentaenoic acid, docosahexaenoic acid, the sum of clupanodonic acid) of fat acid is
59.9%。
Claims (7)
1. a kind of preparation method of the microbial oil rich in phosphatide type polyunsaturated fatty acid, includes the following steps: with fragmentation
Chytrid successively follows these steps to operate as fermenting microbe:
(1) fermentation obtains the microbial fermentation solution for being rich in docosahexaenoic acid;
(2) by the microbial fermentation solution after UF membrane, the concentrated broth that water content is 68% is obtained, the 6L concentration is taken to send out
Zymotic fluid, is added the alkali protease of 3ml, adjusts pH value to 8.0, is warming up to 45 DEG C of heat preservation 4h progress broken walls, then with 10L n-hexane
It extracts, obtained extracting solution is filtered with 1um filter stick, and precipitation obtains total 511g microbial oil;
(3) microbial oil described in 450g is taken, 450ml hexane is added, 0 DEG C of processing 2h is cooled to, then in the environment of 0.2MPa
Under, micro-filtration is kept the temperature with 0.2um pore size ceramic membrane, the semisolid that film retains is obtained into 11g rich in phosphatide type through thin film evaporation
The microbial oil of docosahexaenoic acid.
2. a kind of preparation method of the microbial oil rich in phosphatide type polyunsaturated fatty acid, includes the following steps:
Using microballoon algae as fermenting microbe, successively follow these steps to operate:
(1) fermentation obtains the fermentation liquid of the microbial cells rich in eicosapentaenoic acid, and fermentation liquid is rich in after processing
The dry mycelium of eicosapentaenoic acid;
(2) it takes dry mycelium described in 1kg to carry out crushing broken wall with pulverizer, ethyl alcohol that 1kg content is 95% is added as entrainer,
Use supercritical CO2It is 40 DEG C in temperature, pressure extracts 2h under conditions of being 30Mpa, extraction time is 2 times, the extraction that will be obtained
Liquid is filtered with 1um filter stick, then at 40 DEG C, isolated 357g microbial oil under conditions of 25Mpa;
(3) microbial oil described in 300g is taken, supercritical CO is used2Extraction, extracts 5h under the conditions of temperature 50 C, pressure 35Mpa,
So at 50 DEG C, the isolated 19g of the condition of 28MPa is rich in the arachidonic microbial oil of phosphatide type.
3. a kind of preparation method of the microbial oil rich in phosphatide type polyunsaturated fatty acid, includes the following steps:
Using dino flagellate as fermenting microbe, successively follow these steps to operate:
(1) fermentation obtains the microbial fermentation solution for being rich in docosahexaenoic acid;
(2) above-mentioned fermentation liquid is centrifuged, obtains the concentrated broth that water content is 73%, take the 50L concentrated broth, is added
50ml alkali protease, adjusting pH value to 8.0 are warming up to 50 DEG C of heat preservation 3h and carry out broken walls, then extracted with 100L n-hexane,
Extracting solution is filtered with 1um filter stick, and precipitation obtains total 4168g microbial oil;
(3) microbial oil described in 3kg is taken, 9000ml n-hexane is added, 200g pure water, at 35-40 DEG C, 0.15MPa's is added
Under environment, with 0.2um pore size ceramic membrane micro-filtration, the semisolid that film retains is obtained into 230g rich in phosphatide type through thin film evaporation
The microbial oil of docosahexaenoic acid.
4. a kind of preparation method of the microbial oil rich in phosphatide type polyunsaturated fatty acid, includes the following steps:
Using Mortierella alpina as fermenting microbe, successively follow these steps to operate:
(1) fermentation obtains the fermentation liquid rich in arachidonic microbial cells and is contained after handling the fermentation liquid
Arachidonic dry mycelium;
(2) dry mycelium described in 4kg is taken, 25L n-hexane is added, with colloid mill circulation shear extraction, extracting solution 1um filter stick
Filtering, precipitation obtain total 2128g microbial oil;
(3) 4h is stood in the environment of taking microbial oil described in 2000g to be placed in -10 DEG C, then normal temperature unfreezing, it is clear to pour out upper layer
Oil obtains the thick concentrated phosphatide of 205g;
(4) thick concentrated phosphatide described in above-mentioned 205g is warming up to 65 DEG C, the food-grade H that 4g concentration is 30% is added2O2It is stirred to react
30min then heats to 85-90 DEG C, and residual moisture and H are removed in the environment that vacuum degree is -0.095Mpa2O2, small to moisture
After 0.5%, 50 DEG C are cooled to hereinafter, obtaining 193g rich in the arachidonic microbial oil of phosphatide type.
5. a kind of preparation method of the microbial oil rich in phosphatide type polyunsaturated fatty acid, includes the following steps:
To produce linolenic saccharomycete as fermenting microbe, successively follow these steps to operate:
(1) fermentation obtains the fermentation liquid for being rich in linolenic microbial cells, and fermentation liquid is obtained after processing rich in linolenic acid
Dry mycelium;
(2) dry mycelium described in 9kg is taken, with Ultrasonic Pulverization, 90L n-hexane is added, extracting solution is filtered with 1um filter stick, and precipitation obtains
Total 3978g microbial oil;
(3) microbial oil described in 3600g is taken, vapor is passed through to 105 DEG C of processing 5min, is then cooled to 80 DEG C of centrifugations and obtains
The thick concentrated phosphatide of 918g;
(4) by the thick concentrated phosphatide of 850g in 85-90 DEG C, vacuum degree -0.095Mpa removes the moisture in raw phospholipid, in raw phospholipid
After moisture is less than 1%, 65 DEG C are cooled to, 1800ml n-hexane, 25g active carbon is then added, 50 DEG C of decoloration 30min of temperature control take out
Obtained filtrate thin film evaporator precipitation is filtered, obtains 396g rich in the linolenic microbial oil of phosphatide type.
6. a kind of preparation method of the microbial oil rich in phosphatide type polyunsaturated fatty acid, includes the following steps:
Using Dunaliella salina as fermenting microbe, successively follow these steps to operate:
(1) fermentation obtains the fermentation liquid for being rich in linolenic microbial cells, after handling the fermentation liquid, obtains and contains flax
The dry mycelium of acid;
(2) dry mycelium described in 50kg is taken, 250L n-hexane is added, shears extraction, extracting solution 1um with hand-held cutter
Filter stick filtering, precipitation obtain total 12.9kg microbial oil;
(3) microbial oil described in 10kg is taken, is warming up to 80 DEG C, 1kg pure water is added and carries out hydration reaction 30min, is then centrifuged for
Obtain the thick concentrated phosphatide of 2.81kg;
(4) it by thick concentrated phosphatide described in 2.75kg in 85-90 DEG C, is removed in raw phospholipid in the environment of vacuum degree -0.095Mpa
Moisture be cooled to 65-70 DEG C after moisture in raw phospholipid is less than 1%, the food-grade H that 10ml concentration is 30% is added2O2It stirs
Reaction 30min is mixed, then heats to 85-90 DEG C, residual moisture and H are removed in the environment of vacuum degree -0.095Mpa2O2, to water
After dividing less than 0.5%, 50 DEG C are cooled to hereinafter, obtaining 1.97kg rich in the linolenic microbial oil of phosphatide type.
7. a kind of preparation method of the microbial oil rich in phosphatide type polyunsaturated fatty acid, includes the following steps:
Using schizochytrium limacinum as fermenting microbe, successively follow these steps to operate:
(1) fermentation obtains the fermentation liquid of the microbial cells rich in docosahexaenoic acid, and fermentation liquid is obtained richness after processing
Dry mycelium containing docosahexaenoic acid;
(2) it takes dry mycelium described in 500kg to carry out crushing broken wall with pulverizer, 2500L n-hexane is added and extracts, extracting solution 1um
Then filter stick filtering obtains total 247.3kg microbial oil through vacuum desolvation, the cold acetone removing grease of 800L, weight is added
2 times multiple, obtained wet solid is vacuum dried, removes acetone, obtains 7.7kg powder phospholipid.
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CN110074211A (en) * | 2019-06-05 | 2019-08-02 | 济南极源生物科技有限公司 | A kind of preparation method of the product containing antarctic krill oil and the product being prepared |
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