CN104962589A - Microbial oil rich in phospholipid type polyunsaturated fatty acid and preparation method thereof - Google Patents
Microbial oil rich in phospholipid type polyunsaturated fatty acid and preparation method thereof Download PDFInfo
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Abstract
The invention provides a preparation method of microbial oil rich in phospholipid type polyunsaturated fatty acid. The preparation method comprises the following steps of: (1) fermenting to obtain a microbe rich in polyunsaturated fatty acid; (2) carrying out wall-breaking extraction to obtain the microbial oil; (3) gathering phospholipid contained in the microbial oil, and purifying to obtain the microbial oil rich in the phospholipid type polyunsaturated fatty acid. The preparation method provided by the invention has the beneficial effects of being unlimited in raw material because the whole preparation method takes the naturally sourced microbial oil as the raw material, achieving high content of the polyunsaturated fatty acid contained in the prepared phospholipid type polyunsaturated fatty acid microbial oil, and being safer and wider in application scope compared with the traditional phospholipid type polyunsaturated fatty acid synthesized through an enzymatic method.
Description
Technical field
The present invention relates to and be a kind ofly rich in microbial oil of phosphatide type polyunsaturated fatty acid and preparation method thereof.
Background technology
Phosphatide is the lipoid cpd containing phosphoric acid, is prevalent in biological cell matter and cytolemma, plays a crucial role to maintenance cell membrane function and then maintenance cellular metabolism.Phosphatide can be divided into glyceric alcohol phosphatides and the large class of sphingophospholipid two by its molecular structure.Glyceric alcohol phosphatides mainly comprises Yelkin TTS, kephalin, serine phosphatide, lipositol etc.For the disease that nervous system function lowly brings, as lather, poor appetite, tinnitus, sexual dysfunction power, nervosa diarrhea, practise perseverance constipation and neural pass on material want the dementia brought and internal memory power go down, every day supplements Yelkin TTS fully, can receive good curative effect.Studies have found that the rat raising pregnancy with Yelkin TTS, its offspring is in intellectual check (labyrinth test), and memory is significantly better than the offspring of the rat not raising Yelkin TTS.7th Yelkin TTS international conference in 1997 was once drawn a conclusion: " suggestion pregnant woman takes appropriate Yelkin TTS, and this intelligent growth for child is very important." food and agricultural organization of Yelkin TTS united state (FAO) was classified as one of large nutritive food in the world five 1998 years; U.S. foods in 1999 and the medication management council (FDA) clear stipulaties, all will supplement Yelkin TTS in all baby's recipes in right amount.
Current phosphatide is mainly derived from plant, and commercially available phosphatide mainly contains saturated fatty acid or the monounsaturated fatty acids of 16 or 18 carbon atoms containing palmitinic acid, oleic acid, linolic acid and stearic acid etc.But except these lipid acid, human body also needs supplementary polyunsaturated fatty acid, as linolenic acid, arachidonic acid, timnodonic acid, clupanodonic acid and docosahexenoic acid etc.Docosahexenoic acid in the brain dense, is considered to play vital effect in the development of vision, cognitive function and other normal brain functions.Arachidonic acid is the abundantest in mammalian body, also be most active a kind of polyunsaturated fatty acid, it not only has very important meaning to the health of human body, also most important to the normal development of infant, arachidonic acid also has preventing cardiovascular disease, immunomodulatory, auxiliary Tumor suppression, prevention canceration and regulates neural function, can the disease such as prevention and therapy hypertension, diabetes, obesity and virus infection.
Compared with methyl esters/ethyl ester type or triglyceride type polyunsaturated fatty acid, phosphatide type polyunsaturated fatty acid not only has that bioavailability is high, security is high and the advantage such as good stability, simultaneously for body provides abundant phosphatide, the effect of 1+1>2 can also be played to the growth of brain.There are some researches show, the digestibility of human body to ethyl ester type docosahexenoic acid only has 21%, and to the digestibility of phosphatide type docosahexenoic acid more than 99%.Docosahexenoic acid in yolk and the Yelkin TTS in yolk are closely linked, digested together with Yelkin TTS, are particularly suitable for pregnant and lying-in women and infants.
China Patent Publication No. is that the patent of invention of CN103509047A discloses a kind of extraction process of phosphatidylcholine of antarctic krill and the preparation method of phosphatidylserine.The method is 7.6-8.1 % by collecting krill head and stirring the series of process acquisition docosahexenoic acid content such as extraction by decoction, and Content of Eicosapentaenoic Acid is the phospholipid prod of 14.6-15.6%.The drawback of this method is: the polyunsaturated fatty acid that the operations such as raw material sources has larger restriction, decoction meeting loss is a large amount of, and can damage environment without restraint fishing for of krill.
China Patent Publication No. is that the patent of invention of CN101195637B discloses and a kind ofly prepares the technique being rich in polyunsaturated fatty acid phosphatide, the method carries out transesterify catalyzed reaction mainly through enzyme to soybean lecithin and the free fatty acids being rich in timnodonic acid and docosahexenoic acid, thus obtains the phosphatide being rich in polyunsaturated fatty acid.But in the phosphatide that this method is obtained, timnodonic acid and docosahexenoic acid content lower.And this method needs to use zymin, therefore the range of application of finished product can be restricted.
To sum up, provide a kind of useful environment, Sustainable development, have wider range of application to be rich in microbial oil of phosphatide type polyunsaturated fatty acid and preparation method thereof real in necessary.
Summary of the invention
The object of this invention is to provide a kind of preparation method of microbial oil that can meet the Production requirement of industrialization, that be rich in phosphatide type polyunsaturated fatty acid.
Another object of the present invention is to provide one and has high-content phosphatide, and has the microbial oil of high-content polyunsaturated fatty acid in phosphatide.
For realizing above-mentioned main purpose, the invention provides a kind of preparation method being rich in the microbial oil of phosphatide type polyunsaturated fatty acid, comprise the steps: that (1) fermentation obtains the microorganism of 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 being rich in phosphatide type polyunsaturated fatty acid after purifying.
The preparation method being rich in the microbial oil of phosphatide type polyunsaturated fatty acid of the present invention has following beneficial effect: 1, whole preparation method with the microbial oil of natural origin for raw material, therefore raw material is unrestricted, 2, in the phosphatide type polyunsaturated fatty acid microbial oil prepared, content of polyunsaturated fatty acid is high, 3, compared with the phosphatide type polyunsaturated fatty acid of existing enzymatic clarification, safer, range of application is more extensive.
Further, in step (3), implement enrichment with stand at low temperature, solvent elution, supercritical extraction, membrane separation process, water or steam.The means of these enrichments meet the relevant regulations of food safety.
For realizing another object of the present invention, the invention provides a kind of microbial oil being rich in phosphatide type polyunsaturated fatty acid, in described microbial oil, phospholipids content is not less than 38%, and in phosphatide, the content of polyunsaturated fatty acid is not less than 30%.
The microbial oil being rich in phosphatide type polyunsaturated fatty acid of the present invention has following beneficial effect: first, and the polyunsaturated fatty acid in grease is microbe-derived, therefore, it is possible to adapt to large-scale Production requirement, and edible safer; Secondly, the phospholipids content in grease is high, and in phosphatide, the content of polyunsaturated fatty acid is also high, be more conducive to be absorbed, and nutritive ingredient is high.
Further, be above-mentionedly rich in the microbial oil of phosphatide type polyunsaturated fatty acid, in phosphatide, content of polyunsaturated fatty acid is 30%-79%.
Further, be above-mentionedly rich in the microbial oil of phosphatide type polyunsaturated fatty acid, polyunsaturated fatty acid comprises linolenic acid, arachidonic acid, timnodonic acid, clupanodonic acid and docosahexenoic acid.
Further, be above-mentionedly rich in the microbial oil of phosphatide type polyunsaturated fatty acid, the source of polyunsaturated fatty acid microbial oil is Mortierella alpina, microballoon algae, yeast, schizochytrium limacinum, Dunaliella salina or dino flagellate.
Embodiment
Below in conjunction with specific examples, the present invention is described in further detail.
Embodiment 1
Using schizochytrium limacinum as fermented bacterium, follow these steps to operation successively:
(1) fermentation obtains the microbial fermentation solution being rich in docosahexenoic acid;
(2) by after above-mentioned fermentation liquor membrane sepn, obtain the concentrated broth that water content is 68%, get this concentrated broth of 6L, add the Sumizyme MP of 3ml, adjust ph to 8.0, is warming up to 45 DEG C of insulation 4h and carries out broken wall, then extract with 10L normal hexane.The extracting solution 1um filter stick obtained filters, and precipitation obtains 511g microbial oil altogether;
(3) 450g mentioned microorganism grease is got, add 450ml hexane, be cooled to 0 DEG C of process 2h, then under the environment of 0.2MPa, with 0.2um pore size ceramic membrane insulation micro-filtration, film is retained the semisolid obtained and obtain through thin film evaporation the microbial oil that 11g is rich in phosphatide type docosahexenoic acid.This step is the technique of simultaneously phosphatide being carried out to enrichment and purification process.After testing, the phospholipids content of this microbial oil is 69.7%, and in phosphatide, the total content (eicosatetraenoic acid, timnodonic acid, docosahexenoic acid, clupanodonic acid sum) of polyunsaturated fatty acid is 79%.
Embodiment 2
Using microballoon algae as fermented bacterium, follow these steps to operation successively:
(1) fermentation obtains the fermented liquid being rich in the microbial cells of timnodonic acid, fermented liquid is obtained after treatment the dry mycelium being rich in timnodonic acid;
(2) get the above-mentioned dry mycelium pulverizer of 1kg and carry out pulverize and break cellular wall, add 1kg content be the ethanol of 95% as entrainment agent, use supercritical CO
2be 40 DEG C in temperature, pressure is extract 2h under the condition of 30Mpa, and extraction time is 2 times.The extracting solution 1um filter stick obtained is filtered, then at 40 DEG C, is separated under the condition of 25Mpa and obtains 357g microbial oil;
(3) get 300g mentioned microorganism grease, use supercritical CO
2extraction, at temperature 50 C, extracts 5h under pressure 35Mpa condition, and so at 50 DEG C, the condition separation of 28MPa obtains 19g and is rich in the arachidonic microbial oil of phosphatide type.This step is the technique of simultaneously phosphatide being carried out to enrichment and purification process.After testing, in this microbial oil, phospholipids content is 41.8%, and in phosphatide, the total content (arachidonic acid and timnodonic acid sum) of polyunsaturated fatty acid is 38.1%.
Embodiment 3
Using dino flagellate as fermented bacterium, follow these steps to operation successively:
(1) fermentation obtains the microbial fermentation solution being rich in docosahexenoic acid;
(2) by centrifugal for above-mentioned fermented liquid; obtain the concentrated broth that water content is 73%; get this concentrated broth of 50L; add 50ml Sumizyme MP; adjust ph to 8.0, is warming up to 50 DEG C of insulation 3h and carries out broken wall, then extract with 100L normal hexane; extracting solution 1um filter stick filters, and precipitation obtains 4168g microbial oil altogether;
(3) 3kg mentioned microorganism grease is got, add 9000ml normal hexane, add 200g pure water, at 35-40 DEG C, under the environment of 0.15MPa, with 0.2um pore size ceramic membrane micro-filtration, film is retained the semisolid obtained and obtain through thin film evaporation the microbial oil that 230g is rich in phosphatide type docosahexenoic acid.This step is the technique of simultaneously phosphatide being carried out to enrichment and purification process.After testing, the phospholipids content of this microbial oil is 58%, and in phosphatide, the total content (docosahexenoic acid and timnodonic acid sum) of polyunsaturated fatty acid is 43.1 %.
Embodiment 4
Using Mortierella alpina as fermented bacterium, follow these steps to operation successively:
(1) fermentation obtains the fermented liquid being rich in arachidonic microbial cells, after processing, obtains containing arachidonic dry mycelium to this fermented liquid;
(2) get 4kg dry mycelium, add 25L normal hexane, with colloidal mill circulation shear extraction, extracting solution 1um filter stick filters, and precipitation obtains 2128g microbial oil altogether;
(3) get under 2000g mentioned microorganism grease is placed in the environment of-10 DEG C and leave standstill 4h, then normal temperature unfreezing, pour out upper strata edible vegetable oil, obtain the thick concentrated phosphatide of 205g.This step is the process of enriched microorganism phospholipid in lipid;
(4) thick for above-mentioned 205g concentrated phosphatide is warming up to 65 DEG C, adds the food grade H that 4g concentration is 30%
2o
2stirring reaction 30min, is then warming up to 85-90 DEG C, is to remove residual moisture and H in the environment of-0.095Mpa in vacuum tightness
2o
2.After moisture is less than 0.5%, be cooled to less than 50 DEG C, obtain 193g and be rich in the arachidonic microbial oil of phosphatide type.After testing, the phospholipids content of this microbial oil is 38%, and in phosphatide, the total content (linolenic acid and arachidonic acid sum) of polyunsaturated fatty acid is 59.1 %.
Embodiment 5
To produce linolenic yeast as fermented bacterium, follow these steps to operation successively:
(1) fermentation obtains the fermented liquid being rich in linolenic microbial cells, is obtained after treatment by fermented liquid and is rich in linolenic dry mycelium;
(2) get 9kg dry mycelium, by ultrasonication, add 90L normal hexane, extracting solution 1um filter stick filters, and precipitation obtains 3978g microorganism altogether;
(3) get 3600g mentioned microorganism grease, pass into water vapour to 105 DEG C process 5min, be then cooled to 80 DEG C and centrifugally obtain the thick concentrated phosphatide of 918g.This step is the phosphatide in enriched microorganism grease.
(4) by thick for 850g concentrated phosphatide in 85-90 DEG C, vacuum tightness-0.095Mpa removes the moisture in raw phospholipid.After moisture is less than 1% in raw phospholipid, be cooled to 65 DEG C.Then 1800ml normal hexane is added, 25g gac, temperature control 50 DEG C decolouring 30min.The filtrate that suction filtration obtains uses thin-film evaporator precipitation, obtains 396g and is rich in the linolenic microbial oil of phosphatide type.After testing, in this microbial oil, phospholipids content is 63.7%.And the content of polyunsaturated fatty acid (i.e. linolenic acid) is 30% in phosphatide.
Embodiment 6
Using Dunaliella salina as fermented bacterium, follow these steps to operation successively:
(1) fermentation obtains the fermented liquid being rich in linolenic microbial cells, after processing, obtains containing linolenic dry mycelium to this fermented liquid;
(2) get 50kg dry mycelium, add 250L normal hexane, shear extraction with hand-held shears, extracting solution 1um filter stick filters, and precipitation obtains 12.9kg microbial oil altogether;
(3) get 10kg mentioned microorganism grease, be warming up to 80 DEG C, add 1kg pure water and carry out hydration reaction 30min, then centrifugally obtain the thick concentrated phosphatide of 2.81kg.This step is the technique of the phosphatide in enriched microorganism grease.
(4) by thick for 2.75kg concentrated phosphatide in 85-90 DEG C, under the environment of vacuum tightness-0.095Mpa, remove the moisture in raw phospholipid.After moisture is less than 1% in raw phospholipid, be cooled to 65-70 DEG C, add the food grade H that 10ml concentration is 30%
2o
2stirring reaction 30min, is then warming up to 85-90 DEG C, under the environment of vacuum tightness-0.095Mpa, remove residual moisture and H
2o
2,after moisture is less than 0.5%, be cooled to less than 50 DEG C, obtain 1.97kg and be rich in the linolenic microbial oil of phosphatide type.After testing, in this microbial oil, phospholipids content is 61.1%, and in phosphatide, the total content (linolenic acid) of polyunsaturated fatty acid is 39.4%.
Embodiment 7
Using schizochytrium limacinum as fermented bacterium, follow these steps to operation successively:
(1) fermentation obtains the fermented liquid being rich in the microbial cells of docosahexenoic acid, fermented liquid is obtained after treatment the dry mycelium being rich in docosahexenoic acid;
(2) get 500kg dry mycelium pulverizer and carry out pulverize and break cellular wall, add 2500L normal hexane extraction, extracting solution 1um filter stick filters, and then obtain 247.3kg microbial oil altogether through vacuum desolvation, the cold acetone adding 800L removes grease, repeats 2 times.The wet solid obtained is through vacuum-drying, and removing acetone, obtain 7.7kg powder lecithin, the powder lecithin of the present embodiment is a kind of specific form of the microbial oil being rich in phosphatide type polyunsaturated fatty acid.This step is purification step, and main purpose removes the impurity such as grease, solvent.After testing, in this microbial oil, phospholipids content is 96.7%, and in phosphatide, the total content (eicosatetraenoic acid, timnodonic acid, docosahexenoic acid, clupanodonic acid sum) of polyunsaturated fatty acid is 59.9%.
Claims (8)
1. be rich in a preparation method for the microbial oil of phosphatide type polyunsaturated fatty acid, comprise the steps:
(1) fermentation obtains the microorganism of being rich in polyunsaturated fatty acid,
(2) extraction is carried out to microorganism, obtains microbial oil,
(3) phosphatide in enriched microorganism grease, obtains the microbial oil being rich in phosphatide type polyunsaturated fatty acid after purifying.
2. a kind of preparation method being rich in the microbial oil of phosphatide type polyunsaturated fatty acid according to claim 1, is characterized in that: in step (3), implements enrichment with stand at low temperature, solvent elution, supercritical extraction, membrane separation process, water or steam.
3. a kind of preparation method being rich in the microbial oil of phosphatide type polyunsaturated fatty acid according to claim 1, is characterized in that: described polyunsaturated fatty acid comprises linolenic acid, arachidonic acid, timnodonic acid, clupanodonic acid and docosahexenoic acid.
4. be rich in a microbial oil for phosphatide type polyunsaturated fatty acid, it is characterized in that: in described microbial oil, phospholipids content is not less than 38%, and in phosphatide, the content of polyunsaturated fatty acid is not less than 30%.
5. a kind of microbial oil being rich in phosphatide type polyunsaturated fatty acid according to claim 4, is characterized in that: in phosphatide, content of polyunsaturated fatty acid is 30%-79%.
6. a kind of microbial oil being rich in phosphatide type polyunsaturated fatty acid according to claim 4, is characterized in that: described polyunsaturated fatty acid comprises linolenic acid, arachidonic acid, timnodonic acid, clupanodonic acid and docosahexenoic acid.
7. a kind of microbial oil being rich in phosphatide type polyunsaturated fatty acid according to claim 4, is characterized in that: the source of described polyunsaturated fatty acid microbial oil is Mortierella alpina, microballoon algae, yeast, schizochytrium limacinum, Dunaliella salina or dino flagellate.
8. a kind of microbial oil being rich in phosphatide type polyunsaturated fatty acid according to claim 4, is characterized in that: described in be rich in phosphatide type polyunsaturated fatty acid microbial oil by obtained by the arbitrary described preparation method of claim 1-3.
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