CN104982920A - Composition comprising cells and a polyunsaturated fatty acid having at least 20 carbon atoms (LC-PUFA) - Google Patents

Composition comprising cells and a polyunsaturated fatty acid having at least 20 carbon atoms (LC-PUFA) Download PDF

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Publication number
CN104982920A
CN104982920A CN201510252839.9A CN201510252839A CN104982920A CN 104982920 A CN104982920 A CN 104982920A CN 201510252839 A CN201510252839 A CN 201510252839A CN 104982920 A CN104982920 A CN 104982920A
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pufa
composition
seed
oil
preferred
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丹尼尔·韦尔科埃尔简
亨德里克·路易斯·比杰尔
克里斯蒂安·左尔
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DSM IP Assets BV
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DSM IP Assets BV
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Priority claimed from CN2010800499791A external-priority patent/CN102665431A/en
Publication of CN104982920A publication Critical patent/CN104982920A/en
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
    • A23C9/1528—Fatty acids; Mono- or diglycerides; Petroleum jelly; Paraffine; Phospholipids; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23D—EDIBLE OILS OR FATS, e.g. MARGARINES, SHORTENINGS, COOKING OILS
    • A23D9/00—Other edible oils or fats, e.g. shortenings, cooking oils
    • A23D9/007—Other edible oils or fats, e.g. shortenings, cooking oils characterised by ingredients other than fatty acid triglycerides
    • 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
    • C11B5/00—Preserving by using additives, e.g. anti-oxidants
    • C11B5/0085—Substances of natural origin of unknown constitution, f.i. plant extracts
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/12—Unicellular algae; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
    • C12N15/8243—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
    • C12N15/8247—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine involving modified lipid metabolism, e.g. seed oil composition
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Abstract

The present invention relates to a composition comprising a polyunsaturated fatty acid having at least 20 carbon atoms (LC-PUFA) and cells, which composition has a thermal induction time (T.I.T.) of >24 hours at 40 DEG C. The invention also relates to a process for drying a composition comprising cells and a LC-PUFA, the process comprising drying the composition at a temperature of below 40 DEG C.

Description

Comprise cell and there is the composition of polyunsaturated fatty acid (LC-PUFA) of at least 20 carbon atoms
The present patent application is on November 02nd, 2010 based on the applying date, application number is " 201080049979.1 " (international application no is PCT/EP2010/066598), the divisional application of the application for a patent for invention that name is called " comprise cell and have the composition of polyunsaturated fatty acid (LC-PUFA) of at least 20 carbon atoms ".
Technical field
The present invention relates to a kind of composition of polyunsaturated fatty acid (LC-PUFA) comprising cell and there are at least 20 carbon atoms, relate to a kind of method comprising the composition of cell and LC-PUFA for drying, relate to a kind of method obtaining LC-PUFA or the oil containing LC-PUFA from the composition comprising cell and LC-PUFA.
Background technology
LC-PUFA can be produced by fermentation method by microorganism.LC-PUFA also can produce in plant.Subsequently, the microorganism containing LC-PUFA or plant part can be pretreated, can isolate LC-PUFA or the oil containing LC-PUFA afterwards.
Such as, WO 2006/085672 describes a kind of method, and wherein LC-PUFA is separated from microbial biomass.Wet cell is dry in two-stage drying technology.120 DEG C or higher baking temperature can be used, obtain the dried cellular that water capacity is 1-2wt.%.
After producing the composition containing LC-PUFA, can therefrom be separated LC-PUFA and/or the oil containing LC-PUFA immediately.But the composition containing LC-PUFA in reality was usually stored and/or transported before using (being such as separated LC-PUFA and/or the oil containing LC-PUFA) further.
Have found that, the composition susceptible containing LC-PUFA is in Automatic-heating.That is, between the storage life, temperature may spontaneously increase, and finally causes less desirable blast and fire.Also find, this neurological susceptibility increases with the increase of LC-PUFA content, increases with the increase of the double bond quantity of LC-PUFA.
Target of the present invention is to provide the composition that one comprises (i) LC-PUFA and (ii) cell, and said composition is safer.
Summary of the invention
The present invention now provided with a kind of composition, it comprises: (i) has polyunsaturated fatty acid (LC-PUFA) and (ii) cell of at least 20 carbon atoms, and the thermal induction time (T.I.T.) of said composition at 40 DEG C is greater than 24 hours.
Advantage according to composition of the present invention is, its security is improved, that is, its autoignition temperature increases, and decreases less desirable blast and fire.Another advantage of composition of the present invention is, the storage of composition negatively can not affect the quality of LC-PUFA or the oil containing LC-PUFA, or is at least less to the degree of qualitative effects.
The thermal induction time (T.I.T.) of composition according to the present invention at 40 DEG C is greater than 24 hours.The T.I.T. of composition is by using as Recommendations on the Transport of DangerousGoods, Manual of Tests and Criteria, Section 28.4.4, Test H.4, United Nations, New York, the accumulation of heat described in 1999 stores method of testing and measures, shown in wherein adjusting and providing as follows: the glass Dewar (Dewar vessel) that use internal diameter is 57mm, height is the 0.5L of 210mm.The heat loss of Dewar bottle is 16mW/K.Sample size accounts for 75% of Dewar bottle volume.With Dewar bottle on the rubbery plug lid being highly about 50mm, loosely jam-pack is to allow to breathe.By the hole of central plug, thermocouple is inserted in Dewar bottle.The Dewar bottle of the sample containing initial temperature being 20 DEG C is put into the chamber that control is 40 DEG C, use thermocouple to monitor the temperature of sample.The thermal induction time is defined as: elapsed time between the temperature of sample reaches moment that the moment of 2 DEG C (namely 38 DEG C) lower than the temperature of chamber and the temperature of sample reach higher than the temperature of chamber 2 DEG C (namely 42 DEG C).Fig. 1 describes the mensuration of T.I.T..
Particularly, the present invention relates to every as follows:
1. a composition, it comprises: (i) has the polyunsaturated fatty acid (LC-PUFA) of at least 20 carbon atoms, (ii) cell, the thermal induction time (T.I.T.) of said composition at 40 DEG C is greater than 24 hours.
2. the composition as described in item 1, its water capacity is 1-20wt.%, is preferably 2-15wt.%, is preferably 3-12wt.%, is preferably 3.5-10wt.%, is preferably 4-9wt.%.
3. the composition as described in item 1 or 2, its oil content is at least 10wt.%, such as at least 20wt.%, such as at least 30wt.%, such as at least 40wt.%, such as, lower than 70wt.%, such as, lower than 60wt.%.
4. as the composition above as described in any one, wherein said composition comprises such oil, wherein based on the amount of aliphatic acid all in described oil, described oil contains the PUFA with at least 3 double bonds of at least 10wt.%, such as at least 20wt.%, such as at least 30wt.%, such as at least 40wt.%; Based on the amount of aliphatic acid all in described oil, described oil contain such as lower than 80wt.%, such as lower than 70wt.%, such as lower than the PUFA with at least 3 double bonds of 60wt.%.
5., as the composition above as described in any one, wherein said LC-PUFA has at least 3 double bonds.
6., as the composition above as described in any one, wherein said LC-PUFA is ω-3 or ω-6PUFA.
7. as the composition above as described in any one, wherein said LC-PUFA is selected from by dihomo-gamma-linolenic acid (DGLA, 20:3 ω-6), arachidonic acid (ARA, 20:4 ω-6), eicosapentaenoic acid (EPA, 20:5 ω-3), DHA (DHA:22:6 ω-3), clupanodonic acid (DPA, 22:5 ω-3, or DPA 22:5, ω-6) group that forms.
8., as the composition above as described in any one, wherein said LC-PUFA is ARA or DHA.
9., as the composition above as described in any one, wherein said cell is microbial cell.
10. the composition as described in item 9, wherein said microbial cell is yeast cells, bacterial cell, fungal cell or alga cells.
11. as the composition above as described in any one, and wherein said composition comprises the microorganism that Mortierella belongs to, and preferably comprise the microorganism of Mortierella alpina kind, wherein preferably, described LC-PUFA is ARA.
12. according to any one of item 1-10 composition, wherein said composition comprises Thraustochytriales object microorganism, the microorganism that such as Thraustochytrium belongs to or Schizochytrium belongs to, wherein preferably, described LC-PUFA is DHA or EPA.
13. according to any one of item 1-10 composition, wherein said composition comprise Crypthecodinium belong to microorganism, preferably comprise the microorganism of Crypthecodinium cohnii kind, wherein preferably, described LC-PUFA is DHA.
14. according to any one of item 1-8 composition, wherein said cell is plant cell.
15. compositions as described in item 14, wherein said cell is the plant cell of genetically modified plants.
16. compositions as described in item 14 or 15, wherein said cell is the plant cell of crucifer, preferably the plant cell of plant that belongs to of Brassica
17. 1 kinds of methods comprising the composition of cell and LC-PUFA for drying, described method comprises, lower than 40 DEG C, preferably lower than 35 DEG C, preferably lower than 33 DEG C, preferably lower than 30 DEG C, preferably lower than the temperature of 25 DEG C under dry described composition.
18. methods comprising the composition of cell and LC-PUFA for drying as described in item 17, described method comprises, and described composition is contacted with the air through regulating preferably with dew point <15 DEG C, preferably <10 DEG C, preferred <5 DEG C.
19. methods as described in item 17 or 18, it comprises and uses the dry described composition of fluidized bed dryer.
20. by the available composition of method in item 17-19 described in any one.
21. compositions as described in item 20, its have as item 2 the water capacity that defines.
22. 1 kinds for obtaining the method for LC-PUFA or the oil containing LC-PUFA, described method comprises, from according to composition above described in any one or from obtained by the method item 17 or 21 described in any one or available drying composition in be separated LC-PUFA or the oil containing LC-PUFA.
23. 1 kinds for obtaining the method for food, particularly baby formula milk powder, described method comprises, method according to item 22 obtains LC-PUFA or the oil containing LC-PUFA, then described LC-PUFA or the oil containing described LC-PUFA is mixed in described food.
Accompanying drawing explanation
Fig. 1 shows the schematic diagram measuring the thermal induction time (T.I.T.).
Detailed Description Of The Invention
Preferably, when measuring at 40 DEG C, be at least 2 days (48 hours) according to the T.I.T. of composition of the present invention, preferably at least 3 days (72 hours), preferably at least 4 days (96 hours), preferably at least 5 days.When measuring at 40 DEG C, T.I.T can be at least 8 days, such as at least 10 days.T.I.T. the specific upper limit is not had.When measuring at 40 DEG C, T.I.T. can be less than 25 days, such as, be less than 20 days.
The composition with increase T.I.T. according to the present invention can obtain based on instruction provided by the present invention.
Composition can be dry composition.Find, if baking temperature reduces, then T.I.T. increases.Also find, if the double bond quantity of the content of the LC-PUFA in composition or LC-PUFA is higher, be conducive to reducing baking temperature.
Such as, baking temperature can lower than 40 DEG C, preferably lower than 35 DEG C, more preferably less than 33 DEG C, more preferably less than 30 DEG C, more preferably less than 25 DEG C.When using in this article, baking temperature refers to the temperature of product in drier.Such as, if drier is fluidized bed dryer, baking temperature refers to the temperature of bed.
Therefore, present invention also offers a kind of method comprising the composition of cell and LC-PUFA for drying, the method is included in lower than 40 DEG C (preferably lower than 35 DEG C, preferably lower than 33 DEG C, preferably lower than 30 DEG C, preferably lower than 25 DEG C) temperature under dry compositions.
Drying can by any suitable method.Drying can be carried out in any suitable drier.Preferably, use can prevent focus being emerged or make focus emerge minimized drier.In one preferred embodiment, fluidized bed dryer is used to carry out drying.
Find, T.I.T. increases with the minimizing of drying time.
In one preferred embodiment, use through regulate air to carry out drying.Preferably, the dew point <15 DEG C of air used, preferred <10 DEG C, preferred <5 DEG C.The benefit reducing dew point is, under preferred (low) baking temperature, can effectively realize preferred water capacity.
Therefore, present invention also offers a kind of method comprising the composition of cell and LC-PUFA for drying, the method comprises makes composition contact with the air through regulating, the preferred <15 DEG C of dew point of the described air through regulating, preferred <10 DEG C, preferred <5 DEG C.Preferably, under above-mentioned preferred baking temperature, drying is carried out.
Also find, T.I.T. increases with the increase of the water capacity of composition.Preferably, the water capacity of (such as drying) composition is at least 1wt.%, preferably at least 2wt.%, preferably at least 3wt.%, preferably at least 4wt.%.Water capacity does not have the specific upper limit.The water capacity of composition can lower than 20wt.%, such as, lower than 15wt.%, such as, lower than 12wt.%, such as, lower than 10wt.%, such as, lower than 9wt.%.Find, make water capacity be reduced to the microbial stability that can increase composition lower than preferred value.
When using in this article, water capacity calculates on the basis of weight in wet base, that is, the basis of the gross weight of composition (comprising dry, oil and moisture) calculates.It can be measured by those skilled in the art, such as, by making water evaporate at the temperature of 105 DEG C, then measures the weight of the moisture of evaporation.
In the present invention one preferred embodiment, as disclosed herein preferred water capacity is as disclosed herein obtained to the drying of composition.
Also find, in the processing procedure of composition, avoid the formation of free radical that T.I.T. value can be caused to increase.Based on this opinion, those skilled in the art can avoid the step causing forming free radical.Therefore, cell is usually preferably made may to promote that the exposure in the environment that free radical is formed (such as expose at high temperature and/or in oxygen) minimizes.
If cell is microbial cell (advantageously containing the zymotic fluid of these cells), cell will heat to kill the enzyme that may exist in zymotic fluid fully.Preferred heat protocol is those described in WO 97/037032 and WO 2004/001021, and it is incorporated herein by reference.Preferably, the dissolved oxygen content of the zymotic fluid that heat is very low, such as <10ppm, such as <5ppm, such as <2ppm, such as <1ppm.Killing enzyme (particularly using scheme recited above) may cause T.I.T. value to increase.
In one preferred embodiment, be at least 10wt.% according to the oil content of composition of the present invention, such as at least 20wt.%, such as at least 30wt.%, such as at least 40wt.%.Oil content can lower than 70wt.%, such as, lower than 60wt.%.Oil content can be measured by method known to those skilled in the art.By using n-hexane to make the soxhlet extraction (Soxhlet extraction) of solvent for measuring the proper method of the oil content of composition used herein, wherein carry out the water capacity <15wt.% of the composition extracted, and wherein composition and cell are pulverized (to guarantee that all oil all discharges and can be dissolved in solvent from cell).When using in this article, oil content calculates on the basis of dry weight, that is, the basis of total dry weight (comprise dry and oil, but do not comprise moisture) of composition calculates.
In one preferred embodiment, according to the oil content of composition of the present invention as above define, wherein oil composition as below preferred embodiment as described in.
In one preferred embodiment, composition comprises such oil, wherein relative to the amount of aliphatic acid all in described oil, described oil comprises the PUFA with at least 3 double bonds of at least 10wt.%, such as at least 20wt.%, such as at least 30wt.%, such as at least 40wt.%; Relative to the amount of aliphatic acid all in described oil, described oil comprise such as lower than 80wt.%, such as lower than 70wt.%, such as lower than the PUFA with at least 3 double bonds of 60wt.%.When using in this article, the wt.% with the PUFA of at least 3 double bonds refers to the total amount of all PUFA with at least 3 double bonds.
In one preferred embodiment, composition comprises such oil, wherein relative to the amount of aliphatic acid all in described oil, described oil comprises the arachidonic acid (ARA) of at least 10wt.%, such as at least 20wt.%, such as at least 30wt.%, such as at least 40wt.%; Relative to the amount of aliphatic acid all in described oil, described oil comprise such as lower than 80wt.%, such as lower than 70wt.%, such as lower than the ARA of 60wt.%.
In one preferred embodiment, composition comprises such oil, wherein relative to the amount of aliphatic acid all in described oil, described oil comprises the DHA (DHA) of at least 10wt.%, such as at least 20wt.%, such as at least 30wt.%, such as at least 40wt.%; Relative to the amount of aliphatic acid all in described oil, described oil comprise such as lower than 80wt.%, such as lower than 70wt.%, such as lower than the DHA of 60wt.%.
Be soxhlet extraction (Soxhlet extraction) extraction oil from composition by using n-hexane to make solvent as above for measuring the proper method of oil composition used herein, and measure the aliphatic acid composition of the oil extracted.
If oil content and/or double bond quantity relatively high, preferably select lower baking temperature and the shorter time of staying in drier.
Based on instruction herein, higher T.I.T. value can be obtained, even for the high composition of oil content and/or contain high concentration the composition with the oil of the PUFA of at least 3 double bonds also like this.
When using in this article, following abbreviations is used in whole application:
PUFA refers to polyunsaturated fatty acid;
LC-PUFA (long-chain polyunsaturated fatty acid) refers to the PUFA with at least 20 carbon atoms;
HUFA (highly unsaturated fatty acid) refers to the PUFA with at least 3 double bonds;
LC-HUFA (long-chain highly unsaturated fatty acid) refers to the polyunsaturated fatty acid with at least 20 carbon atoms and at least 3 double bonds.
The invention is not restricted to specific LC-PUFA.In an embodiment of the invention, LC-PUFA has at least 3 double bonds.In yet another embodiment of the present invention, LC-PUFA has at least 4 double bonds.Benefit of the present invention concerning even more remarkable the LC-PUFA that double bond quantity increases because the neurological susceptibility of Automatic-heating increases along with the increase of double bond quantity.
LC-PUFA can be ω-3LC-PUFA or ω-6LC-PUFA.
LC-PUFA comprises such as:
Dihomo-gamma-linolenic acid (dihomo-γ-linolenic acid, DGLA, 20:3 ω-6);
Arachidonic acid (ARA, 20:4 ω-6);
Eicosapentaenoic acid (eicosapentaenoic acid, EPA, 20:5 ω-3);
Clupanodonic acid (DPA, 22:5 ω-3, or DPA 22:5, ω-6),
DHA (DHA:22:6 ω-3).
Preferred LC-PUFA comprises arachidonic acid (ARA) and DHA (DHA).Especially, ARA is preferred.
Composition according to the present invention comprises cell.This cell can be any containing LC-PUFA and/or the cell producing LC-PUFA.
In an embodiment of the invention, cell is microbial cell (microorganism).
The example of microbial cell is yeast cells, bacterial cell, fungal cell and alga cells.Preferred fungi, preferred Mucorales object fungi.The example of described fungi is Mortierella, Phycomyces, Blakeslea, Aspergillus, Thraustochytrium, Pythium or Entomophthora.Preferred arachidonic acid (ARA) source is from Mortierella alpina.Algae can be dinoflagellate (dinoflagellate) and/or comprise Porphyridium, Nitschia or Crypthecodinium (such as Crypthecodinium cohnii).Yeast comprises Pichia and belongs to or Saccharomyces genus, such as Pichia ciferrii.Bacterium can be that Propionibacterium belongs to.
In an embodiment of the invention, composition comprises the fungi that Mortierella belongs to, and preferably comprises the fungi of Mortierella alpina kind, and wherein preferably LC-PUFA is ARA or DGLA.
In an embodiment of the invention, composition comprises Thraustochytriales object fungi, and such as Thraustochytrium belongs to or Schizochytrium belongs to, and wherein preferably LC-PUFA is DHA and/or EPA.
In an embodiment of the invention, composition comprises the algae that Crypthecodinium belongs to, and preferably comprises the algae of Crypthecodinium cohnii kind, and wherein preferably LC-PUFA is DHA.
In yet another embodiment of the present invention, cell is plant cell.Cell can be the plant cell of genetically modified plants.
Described by suitable Plants and Seeds have in such as WO 2005/083093, WO 2008/009600 and WO2009/130291, the content of these documents is incorporated herein by reference.In the present invention, operable other plant and seed are open in such as WO 2008/100545, WO 2008/124806, WO2008/124048, WO 2008/128240, WO 2004/071467, WO 2005/059130, and the content of these documents is incorporated herein by reference.Seed can be (transgenosis) soya seeds or (transgenosis) rape seed.Plant can be (transgenosis) bean plant or (transgenosis) rapeseed plant.
In one preferred embodiment, plant is cruciate flower (Brassicaceae) section (transgenosis) plant, such as Brassica belongs to, Camelina belongs to, Melanosinapis belongs to, Sinapis belongs to, Arabidopsis belongs to, such as following genus and kind: Brassica alba, Brassica carinata, Brassica hirta, Brassica napus, Brassicaa rapa ssp., Sinapis arvensis, Brassica juncea, Brassica juncea var.juncea, Brassica juncea var.crispifolla, Brassica juncea var.foliosa, Brassica nigra, Brassicasinapioides, Camelina sativa, Melanosinapis communis, Brassica oleracea or Arabidopsis thaliana.
Composition can be any living beings comprising LC-PUFA.Preferably, composition is obtained by drying means disclosed herein or available (drying) composition.
Composition can be the microbial biomass comprising microorganism and LC-PUFA.Preferred microorganism and LC-PUFA mention above being those.
In the embodiment that the present invention one is possible, comprise and obtained by following method according to the composition of microorganism of the present invention (microbial cell), the method comprises the zymotic fluid (be also referred to as pasteurization or sterilizing) of heating containing microbial cell, microbial cell is dewatered (such as by filter), then dried microorganism cell in the above-mentioned methods.In one preferred embodiment, the microbial cell through dehydration carries out granulation (preferably by extruding pelletization) before the drying.Preferred granulation (such as extruding pelletization) is carried out at lower than the temperature of 25 DEG C.Described by a kind of preferred technique has in WO97/037032, it is incorporated herein by reference.
In an embodiment of the invention, composition comprises the form that seed containing LC-PUFA and/or composition can be seeds, and the thermal induction time (T.I.T.) of described seed at 40 DEG C is greater than 24 hours.Preferably, the seed of plant mentioned above being of seed.
Find, keep the percentage of low damage seed, T.I.T. will be made to increase.
Preferably, the seed being less than 12% is the seed damaged completely, be preferably less than 8%, be preferably less than 5%, be preferably less than 3% seed be the seed damaged completely.
Preferably, the seed being less than 6% is significantly raw seed (distinctly green seed), be preferably less than 4%, be preferably less than 2%, be preferably less than 1% seed be significantly raw seed.
Preferably, the seed being less than 0.5% is the seed through heating, be preferably less than 0.3%, be preferably less than 0.1%, be preferably less than 0.05% seed be seed through heating.
In one preferred embodiment, the seed being less than 8% is the seed damaged completely, and the seed being less than 4% is significantly raw seed, and the seed being less than 0.3% is the seed through heating.Another preferred embodiment in, the seed being less than 5% is the seed damaged completely, and the seed being less than 2% is significantly raw seed, and the seed being less than 0.1% is the seed through heating.Another preferred embodiment in, the seed being less than 3% is the seed damaged completely, and the seed being less than 1% is significantly raw seed, and the seed being less than 0.05% is the seed through heating.
When using in this article, the seed damaged completely, significantly raw seed and the percentage through the seed of heating measure according to the Official Grain Grading Guide.2001of the Canadian GrainCommission (for rape and rapeseed).
There is the seed damaged completely of preferred percent, significantly raw seed and the seed through the seed of heating can be obtained by the suitable selection of seed after harvesting.
In another aspect of this invention, the invention provides the seed comprising LC-PUFA, the percentage of the seed wherein damaged completely, significantly raw seed and/or the seed through heating is as disclosed above.
Preferably, relative to the total amount of aliphatic acid in seed, this seed comprises the LC-PUFA (such as LC-PUFA as described herein) of at least 5wt.%, preferred at least 10wt.%, preferred at least 15wt.%, preferred at least 20wt.%.
Preferably, relative to the total amount of aliphatic acid in seed, this seed comprises the ω-6LC-PUFA of at least 5wt.%, preferred at least 10wt.%, preferred at least 15wt.%, preferred at least 20wt.%.
Preferably, relative to the total amount of aliphatic acid in seed, this seed comprises the ARA of at least 5wt.%, preferred at least 10wt.%, preferred at least 15wt.%, preferred at least 20wt.%.
Preferably, relative to the total amount of aliphatic acid in seed, this seed comprises the ω-3LC-PUFA of at least 5wt.%, preferred at least 10wt.%, preferred at least 15wt.%, preferred at least 20wt.%.
Preferably, relative to the total amount of aliphatic acid in seed, this seed comprises the DHA of at least 5wt.%, preferred at least 10wt.%, preferred at least 15wt.%, preferred at least 20wt.%.
Preferably, based on the total amount of aliphatic acid in seed, this seed comprises the erucic acid being less than 2wt.%, is preferably less than 1wt.%, is preferably less than 0.5wt.%.
According to composition of the present invention before further use and/or processing, can suitably store.
Advantageously, composition lower than 10 DEG C, preferably lower than 5 DEG C, preferably lower than 0 DEG C, preferably lower than subzero 5 DEG C, preferably lower than the temperature of subzero 10 DEG C under store.Storage temperature does not have specific lower limit.Usually, composition stores at higher than the temperature of subzero 30 DEG C.
If composition comprises the form that seed or composition are seeds, preferably, the water capacity of this seed is less than 15wt.%, such as be less than 12wt.%, such as, be less than 10wt.%, such as, be less than 9.5wt.%, such as higher than 6wt.%, such as, higher than 7wt.%, such as, higher than 8wt.%.Water capacity can such as between 6-15wt.%, such as, between 7-12wt.%, such as, between 8-10wt.%.Preferred water capacity can by making seed drying and obtaining as above.
Composition can time period of stored for any suitable.Composition can such as store at least one sky, such as at least 1 week, such as at least 2 weeks, such as at least 1 month, such as at least 3 months.Storage time does not have the specific upper limit.Said composition such as can store and be less than 12 months, such as, be less than 6 months.
The invention still further relates to the method for obtaining LC-PUFA or the oil containing LC-PUFA, the method comprises from composition according to the present invention or obtains or be separated available composition described LC-PU FA or the oil containing LC-PUFA from method according to the present invention.
Lipid LC-PUFA or the oil containing LC-PUFA can by extracting LC-PUFA or containing the oil of LC-PUFA and obtain, preferably by solvent extraction from composition.Any suitable solvent can be used, such as C 1-10arrcostab (such as ethyl acetate or butyl acetate), toluene, C 1-3alcohol (such as methyl alcohol, propyl alcohol), C 3-6alkane (such as hexane) or supercritical fluid (such as liquid CO 2or supercritical propane).Preferably, this solvent is non-polar solven, such as C 3-C 8alkane (preferred hexane) or supercritical fluid (preferred supercritical CO 2or supercritical propane).Described by preferred extraction process has in WO 97/037032.
If composition comprises the form that seed or composition are seeds, so LC-PUFA and/or the oil containing LC-PUFA can be separated by following method.
Seed and the/composition that comprises seed can be crushed or be pressed into thin slice.This contributes to reclaiming LC-PUFA or the oil containing LC-PUFA.Subsequently, the composition crushing and/or press sheet seed and/or comprise this seed can be heated, such as, higher than at the temperature of 60 DEG C.Heating can be carried out at relatively low temperatures.Seed and/or the composition comprising this seed can such as heat at the temperature between 50-90 DEG C, such as, between 60-80 DEG C, preferably heat the time of 2-60 minute, preferred 5-30 minute.If select the temperature raised, so preferably reduce the duration of heating.
Seed and the/composition that comprises seed can heat with two-forty.The method can such as comprise the composition adding heating seed in the following manner or comprise seed, and wherein temperature is being less than 1 minute, being preferably less than 30 seconds, rising to 70 DEG C from 40 DEG C in time of being preferably less than 20 seconds.The method can such as comprise the composition adding heating seed in the following manner or comprise seed, and wherein temperature is being less than 1 minute, is preferably being less than 30 seconds, rises to 100 DEG C in the time being preferably less than 20 seconds from 40 DEG C.
Can comprise according to method of the present invention, by use superheated steam add heating seed and/comprise the composition of seed.Such as can comprise according to method of the present invention, seed is contacted with superheated steam with the/composition that comprises seed.
Preferably, add under method according to the present invention is included in relatively high temperature (such as between 120-160 DEG C) and in the relatively short time heating seed and/comprise the composition of seed.The method can such as comprise, and is less than 8 minutes, such as, is less than 5 minutes, such as, is less than 3 minutes, is such as less than time period of 2 minutes at the temperature making seed and the/composition that comprises seed remain on higher than 120 DEG C (such as lower than 160 DEG C).Make seed and the/composition that comprises seed remain between 120 DEG C and such as (lower than) time period of at least 5 seconds, such as at least 10 seconds at temperature between 160 DEG C.
Preferably, with relatively high speed cool seed and/comprise the composition of seed.Preferably, make seed and the/temperature that comprises the composition of seed being less than 60 minutes, being preferably less than 30 minutes, being reduced to the temperature of 40 DEG C in time period of being preferably less than 15 minutes from maximum temperature.
These schemes can be used alone or combinationally use.Such as, two-forty heating can with seed and the/composition that comprises seed are remained on preferred temperature under relative short ageing and/or with cool combination fast.
Heating is not limited to a certain moment of method.Heating can be carried out before or after pulverizing seed (such as crush or be pressed into thin slice).On the other hand, the invention provides the method for heating the seed containing LC-PUFA, wherein heating this seed by disclosed above.
The fraction of oil can be obtained by squeezing seed or the composition comprising seed.For discharge oil fraction and squeeze seed can by use methods known in the art carry out.Screw press can be used.In one preferred embodiment, the present invention includes use the squeezer (such as screw press) of cooling to squeeze seed or comprise seed composition to discharge oil.
Solvent extraction in the fraction the also had filter press cake that can be obtained by squeezing from above of oil and obtaining.
The purification of oil can comprise come unstuck, refining, decolouring (bleaching) and/or deodorization.These are known steps, can be implemented by those skilled in the art.In one preferred embodiment, deodorizing carries out at lower than the temperature of 200 DEG C, preferably lower than 190 DEG C, preferably lower than 185 DEG C.Deodorization temperature is reduced to and will improves the quality of oil lower than preferred value.
Present invention also offers a kind of for obtaining the method for food (particularly baby formula milk powder), it comprises from composition according to the present invention, to obtain LC-PUFA or the oil containing LC-PUFA, and described LC-PUFA or the oil containing described LC-PUFA is mixed in described food.
Other preferred aspect, embodiment and features are disclosed in claims.
The preferred feature of an embodiment of the invention and/or aspect and characteristic are equally applicable to another embodiment after carrying out necessary correction.When using in this article, the preferred characteristic sum characteristic of LC-PUFA is applicable to all LC-PUFA in all aspects of the present invention and embodiment.
The present invention is further illustrated by reference to following non-limiting examples.
Embodiment
embodiment 1
The zymotic fluid of Mortierella alpina (ferment and obtain after 8 days) pasteurization 1 hour at 70 DEG C.Filter the zymotic fluid through pasteurization, obtain the filter cake that water capacity is 50wt.%.Filter cake crushed at lower than the temperature of 15 DEG C and extrude.It is 7% that extrudate (diameter is 3mm) is dried to water capacity in the continuous fluid bed dryer with three districts.In the firstth district, bed tempertaure is 32 DEG C, and air themperature is 50 DEG C of (T dew point=15 DEG C).
Firstth district: bed tempertaure 32 DEG C, air themperature 50 DEG C of (T dew point=15 DEG C): 45 minutes
Secondth district: bed tempertaure 32 DEG C, air themperature 35 DEG C of (T dew point=1 DEG C): 45 minutes
3rd district: bed tempertaure 15 DEG C, air themperature 15 DEG C of (T dew point=1 DEG C): 30 minutes
The oil content of dry living beings is 39%.Relative to the amount of aliphatic acid all in oil, the content of ARA is 46%.
The thermal induction time (T.I.T.) of surveying at 40 DEG C: 9 days.
comparative experiment A
Repeat above-mentioned fermentation and pasteurization.Use continuous dehydrating machine reclaim wet cell and make it cracked, carry out drying by heated-air drying (hot blast temperature is 120 DEG C) subsequently, making it be dried to water capacity with vibrated fluidized bed is 1wt.%.Dry cell cooling is made by providing the air of room temperature in fluid bed.ARA is the same with embodiment with the content of oil.
The thermal induction time (T.I.T.) of surveying at 40 DEG C: <12 hour.
embodiment 2
From genetically modified Brassica plant, obtain the seed of the arachidonic acid (total amount relative to aliphatic acid) containing 19%, described genetically modified Brassica plant is by using the method described in WO2008009600 to change.
The specification (measuring according to the Official Grain Grading Guide, 2001of the CanadianGrain Commission) of seed is as follows: significantly raw <2%, damages <5% completely.
Use the seed that the dry water capacity of fluidized bed dryer is 17wt.%.Bed tempertaure is 28 DEG C.Use dew point is the air through regulating of 10 DEG C.The water capacity of the seed of drying is 8.5wt.%.Oil content is 35wt.%.
The thermal induction time (T.I.T.) of surveying at 40 DEG C: 14 days.

Claims (10)

1. a composition, it comprises: (i) has the polyunsaturated fatty acid (LC-PUFA) of at least 20 carbon atoms, (ii) cell, said composition is greater than 24 hours the thermal induction time (T.I.T.) of 40 DEG C.
2. composition as claimed in claim 1, its water capacity is 1-20wt.%, is preferably 2-15wt.%, is preferably 3-12wt.%, is preferably 3.5-10wt.%, is preferably 4-9wt.%.
3. composition as claimed in claim 1 or 2, its oil content is at least 10wt.%, such as at least 20wt.%, such as at least 30wt.%, such as at least 40wt.%, such as, lower than 70wt.%, such as, lower than 60wt.%.
4. composition as claimed in any preceding claim, wherein said composition comprises such oil, wherein based on the amount of aliphatic acid all in described oil, described oil contains the PUFA with at least 3 double bonds of at least 10wt.%, such as at least 20wt.%, such as at least 30wt.%, such as at least 40wt.%; Based on the amount of aliphatic acid all in described oil, described oil contain such as lower than 80wt.%, such as lower than 70wt.%, such as lower than the PUFA with at least 3 double bonds of 60wt.%.
5. composition as claimed in any preceding claim, wherein said LC-PUFA has at least 3 double bonds.
6. composition as claimed in any preceding claim, wherein said LC-PUFA is ω-3 or ω-6PUFA.
7. composition as claimed in any preceding claim, wherein said LC-PUFA is selected from by dihomo-gamma-linolenic acid (DGLA, 20:3 ω-6), arachidonic acid (ARA, 20:4 ω-6), eicosapentaenoic acid (EPA, 20:5 ω-3), DHA (DHA:22:6 ω-3), clupanodonic acid (DPA, 22:5 ω-3, or DPA 22:5, ω-6) group that forms.
8. comprise a method for the composition of cell and LC-PUFA for drying, described method comprises, lower than 40 DEG C, preferably lower than 35 DEG C, preferably lower than 33 DEG C, preferably lower than 30 DEG C, preferably lower than the temperature of 25 DEG C under dry described composition.
9. one kind for obtaining LC-PUFA or the method for oil containing LC-PUFA, described method comprises, from according to composition above described in any one claim or from obtained by method according to claim 8 or available drying composition be separated LC-PUFA or the oil containing LC-PUFA.
10. one kind for obtaining the method for food, particularly baby formula milk powder, described method comprises, method according to claim 9 obtains LC-PUFA or the oil containing LC-PUFA, then described LC-PUFA or the oil containing described LC-PUFA is mixed in described food.
CN201510252839.9A 2009-11-03 2010-11-02 Composition comprising cells and a polyunsaturated fatty acid having at least 20 carbon atoms (LC-PUFA) Pending CN104982920A (en)

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