CN101252844A

CN101252844A - Polyunsaturated fatty acid-containing oil product and uses and production thereof

Info

Publication number: CN101252844A
Application number: CNA2006800321627A
Authority: CN
Inventors: 贾奥德·菲希塔利; 尼尔·F·莱宁格; 杰苏斯·R·阿布里尔; 纳泽·阿梅德; S·P·贾纳卡·N·塞纳纳亚科
Original assignee: Martek Biosciences Corp
Current assignee: DSM IP Assets BV
Priority date: 2005-07-01
Filing date: 2006-06-30
Publication date: 2008-08-27

Abstract

The present invention includes a solid fat composition that includes an oil having saturated fat and a microbial oil having a long chain polyunsaturated fatty acid and an emulsifier. In particular, the solid fat composition can have high levels of long chain polyunsaturated fatty acid and low amounts of emulsifiers. In preferred embodiments, the polyunsaturated oil is an unwinterized microbial oil. The invention also relates to methods for making such compositions and food, nutritional, and pharmaceutical products comprising said compositions. The present invention also includes a microbial oil product prepared by extracting an oil-containing fraction comprising at least one LC-PUFA from a microbial biomass, and treating the fraction by a process of vacuum evaporation, wherein the oil product has not been subject to one or more of a solvent winterization step, a caustic refining process, a chill filtration process, or a bleaching process.

Description

The oil product and uses thereof and the preparation method that comprise polyunsaturated fatty acid

Technical field

The present invention relates to oil product that comprises polyunsaturated fatty acid and uses thereof, the purposes in solid-state fat composition (solid fat composition) for example, described composition comprises long-chain polyunsaturated fatty acid and the thickener that comes from microorganism.The method that the present invention also relates to prepare the method for these products and comprise food, nutriment and the pharmaceutical products of described composition.

Background technology

People expect to improve the diet absorption of multiple beneficial nutrient (nutrients).Useful especially nutrient comprises aliphatic acid, for example ω-3 and ω-6 long-chain polyunsaturated fatty acids (long chainpolyunsaturated fatty acids, LC-PUFA).ω-3PUFAs is considered to important diet compound, is used for prevention of arterial sclerosis and coronary heart disease, amelioration of inflammation state and postpones growth of tumour cell.ω-6PUFAs is not only as the structured lipid of human body, and as the multiple inflammatory factor precursor of Prostaglandins and Leukotrienes for example.The ω-3 that one class is important and ω-6PUFAs are long-chain omega-3 and ω-6PUFAs.

Based on the length and the saturation characteristics of carbochain, aliphatic acid is classified.SCFA has 2 to about 6 carbon, and is normally saturated.Medium chain fatty acid has about 6 to about 14 carbon, also is saturated usually.LCFA has 16 to 24 or more a plurality of carbon, can be saturated or undersaturated.In LCFA more, can there be one or more unsaturated points, draw term " monounsaturated " and " polyunsaturated " respectively.In the present invention, especially pay close attention to and have 20 or the long-chain PUFAs (LC-PUFAs) of more a plurality of carbon.

According to the number and the position of two keys in the aliphatic acid,, LC-PUFAs is classified according to the naming rule of generally acknowledging.Based on the position of two keys of the most close fatty acid methyl cardinal extremity, have the LC-PUFAs of two main series or family: ω-3 series has two keys at the 3rd carbon place, and ω-6 series did not have any pair of key before the 6th carbon.Thereby DHA (docosahexaenoic acid, " DHA ") has the chain length of 22 carbon, and 6 two keys originate in the 3rd carbon of methyl end, called after " 22:6n-3 ".Other important ω-3 LC-PUFAs comprises the eicosapentaenoic acid (eicosapentaenoic acid, " EPA ") of called after " 20:5 n-3 " and ω-3 clupanodonic acid (" DPA ") of called after " 22:5 n-3 ".Important ω-6LC-PUFAs comprises the arachidonic acid (arachidonic acid, " ARA ") of called after " 20:4 n-6 " and ω-6 clupanodonic acid (docosapentaenoic acid, " DPA ") of called after " 22:5 n-6 ".

In human body, can not carry out ω-3 and ω-6 essential fatty acid from the beginning or " completely newly " synthetic; Yet when obtaining these essential fatty acids in diet, body can change into LC-PUFAs with these essential fatty acids, and for example DHA and ARA are although efficient is low.ω-3 and ω-6 aliphatic acid all must be the parts that nutrition is taken in, because compare with the 7th carbon atom from molecule ω terminal number, human body can not insert two keys in the place of more close ω end.Thereby all metabolic conversion are all carried out under the situation that does not change the molecule ω end that comprises ω-3 and ω-6 pair key.Therefore, ω-3 and ω-6 acid are independently aliphatic acid families of two classes, because they can not transform in human body mutually.

In the past twenty years, fitness guru has been recommended lower and the diet that polyunsaturated fat is higher of saturated fat (saturated fats).Although many consumers have adopted this suggestion, heart disease, cancer, diabetes and make still constantly stable the increasing of the incidence of disease of multiple other disease of people's weakness.Scientist approves of, and the types and sources of polyunsaturated fat is the same vital with the total amount of fat.Most of common polyunsaturated fat come from phyteral, lack LCFA (ω-3LC-PUFAs) the most especially.In addition, polyunsaturated fat is carried out hydrogenation to obtain artificial fat, this has caused the increase of some health problem, and aggravates the scarcity of some essential fatty acids.In fact, confirmed that multiple medical conditions can benefit by replenishing ω-3.These illnesss comprise acne, allergy, Alzheimer disease, arthritis, atherosclerotic, galactoncus (breast cysts), cancer, cystic fibrosis, diabetes, eczema, hypertension, hyperfunction (hyperactivity), intestinal disease (intestinaldisorder), renal dysfunction (kidney dysfunction), leukaemia and multiple sclerosis.Noticeable ground, the World Health Organization has recommended infant formula (infant formula) should be rich in omega-fatty acid.

Many unsaturates that tradition is used are those many unsaturates that come from vegetable oil, and described vegetable oil comprises a large amount of ω-6 (C18:2 n-6), but comprises ω-3 seldom or do not comprise any ω-3.Although ω-6 and omega-fatty acid all are essential for good health, recommend and to consume them with about 4: 1 equilibrium relation.The main source of ω-3 has Linseed oil (flaxseed oil) and fish oil (fish oil).The quick growth of experiencing Linseed oil and fish oil production in 10 years in past.This oil of two types is considered to the good diet source of ω-3 polyunsaturated fat.Linseed oil does not comprise any EPA, DHA, DPA or ARA, but comprises leukotrienes (C18:3 n-3), and leukotrienes is to make body can make EPA " building block (building block) ".Yet evidence shows that the speed of metabolic conversion may be slow and unstable, and is especially true in those people that go to bits.Type and content that the aliphatic acid of fish oil is formed are quite big, and this depends on concrete species and diet thereof.For example, compare with those wild fishes, the fish of aquaculture often has the omega-fatty acid of lower content.And fish oil brings the risk of finding that comprises environmental contaminants in the fish of being everlasting.In view of the health advantages of these ω-3 and ω-6LC-PUFAs (chain length is greater than 20), people expect to replenish the food that contains these aliphatic acid.

Be known that fluid oil for example fish oil and certain micro-organisms oil (microbial oil), comprise the LC-PUFAs of high-load.Yet because its undersaturated characteristic, these oil are not solid-state in room temperature (promptly 20 ℃), but are in oil form or liquid form.Yet, expectation be in inapplicable some food applications of fluid oil, to use the solid-state form of the oil be rich in PUFA.In order to form solid-state composition, attempted multiple measure.Be used to make the common methods that unsaturated oils solidifies to comprise, these oil carried out partially or completely hydrogenation, thereby obtain semisolid oil.Yet the result of this chemical conversion is that it is saturated that oil becomes, and lost its healthy characteristic.Partially hydrogenated method also causes the formation of trans-fatty acid, and trans-fatty acid has demonstrated has some harmful characteristics.Therefore, owing to use method for hydrogenation to solidify unsaturated oils, the beneficial characteristics of unsaturated oils is replaced by harmful characteristic of extremely not expecting of the formation of saturated oils and trans-fatty acid.Other method comprises mixes unsaturated oils with " firmly " or saturated fat, be semisolid oil thereby make the gained mixture.Again, because the benefit of " health " unsaturated oils has been offset in the existence of sclerosis or saturated fat at least in part.Other method that is used to form the semisolid fat composition of the high-load polyunsaturated fat that can smear comprises, uses particular type emulsifying agent or other thickener, for example fatty alcohol of high-load.Before the present invention, do not exist in the art and have the following food that comprises solid-state or composition that semisolid is fatty or comprise high-load PUFAs: do not contain the saturated fat that external source is added, do not contain the emulsifying agent of the external source interpolation of high-load, and/or do not conform to the thickener that other type is arranged.These compositions and these method for compositions of preparation are high expectations.Further expectation is, the cost effective method of this composition of preparation is provided, and described method relates to uses harmless material, minimum processing procedure and minimum material inventory.

Typically, handle the known fluid oil that comprises high-load LC-PUFAs by a plurality of steps, microbial oil for example, for people or other animals consuming, described a plurality of steps comprise preliminary treatment, precipitation thinner or deodorization, winterization (winterization), alkali refining (caustic refining) (also being known as chemical refining), cold filtration (chill filtration) and bleaching (bleaching).These processes increase the time of preparation of product and cost, and may introduce natural in subtractive process or the unacceptable chemical substance in organic products market.Therefore, need improved oily preparation method, described method is that simplification, low cost and market can be accepted extensively, and still can prepare effectively simultaneously to have the product that can accept organoleptic attribute.

Summary of the invention

The invention provides the method for the solid-state fat composition of preparation, described method comprises: will comprise the oil and at least a emulsifier of saturated fat and microbial oil, and form mixture, described microbial oil comprises at least a LC-PUFA; Make described mixture solidified, form solid-state fat composition.The present invention also provides solid-state fat composition, and described composition comprises the not mixture of winterization microbial oil and emulsifying agent, and described microbial oil comprises LC-PUFA, and wherein said mixture is a solid-state composition in room temperature.

In some embodiments of the inventive method, described oil comprises the LC-PUFA and about 20wt.% saturated fat to about 60wt.% of about 5wt.% to about 70wt.%.

In some embodiments, solid-state fat composition comprises saturated fat.

In some embodiments, saturated fat is not that external source is added, and in other embodiments, saturated fat is that external source is added (added exogenously).In other embodiments, microbial oil be winterization or hydrogenation.

In some embodiments, microbial oil comes from and is selected from following microorganism: the microorganism of genus thraustochytrium (Thraustochytrium), schizochytrium limacinum belong to the microorganism of (Schizochytrium), the microorganism that Althornia belongs to, the microorganism that Aplanochytrium belongs to, the microorganism that Japonochytrium belongs to, the microorganism that Labyrinthula belongs to, the microorganism that Labyrinthuloides belongs to, the microorganism that Crypthecodinium belongs to, and their mixture.In other embodiments, microorganism is selected from the microorganism of genus thraustochytrium, the microorganism that schizochytrium limacinum belongs to, the microorganism that Crypthecodinium belongs to, and their mixture.

In some embodiments, microbial oil comprises that carbon chain lengths is at least 20 or at least 22 LC-PUFA, or has at least three two keys or have the LC-PUFA of at least four two keys.In some embodiments, LC-PUFA comprises DHA or clupanodonic acid or arachidonic acid or eicosapentaenoic acid.In other embodiments, oil comprises at least about the DHA of 50% weight or at least about the DHA of 60% weight.

In some embodiments, solid-state fat composition has the structure (homogeneoustexture) of homogeneous, or is shortening (shortening).

In some embodiments; emulsifying agent is monoglyceride, diglyceride, monoglyceride/diglyceride combination, lecithin, dilactic acid monoglyceride-diglyceride (lactylated mono-diglyceride), polyglycerol ester, sucrose fatty ester, stearyl dilactic acid sodium (sodium steroyl lactylate), stearyl dilactic acid calcium (calcium steroyl lactylate), and their combination.In other embodiments, the amount of emulsifying agent is extremely about 2.0% weight of about 0.01% weight, in other embodiments, is about 0.2% weight of about 0.05% weight.

In some embodiments of the inventive method, the melt temperature of solid-state fat composition is at least about 20 ℃, at least about 30 ℃ or at least about 35 ℃.

In some embodiments of the inventive method, make the step of mixture solidified control the formation of crystal in the solid-state fat composition.In the embodiment of solid-state fat composition, composition comprises crystal, and in some embodiments, crystal comprises β ' crystal (β-prime crystal).In other embodiment of the inventive method or solid-state fat composition, crystal comprises β ' crystal, fat in solid-state fat composition and/or oily be β ' crystalline form at least about 50wt.%, or the fat in solid-state fat composition and/or oily be β ' crystalline form at least about 80wt.%.

In some embodiments of the inventive method, add deep fat and/or emulsifying agent, before mixed step, add deep fat and/or emulsifying agent, or oil and/or emulsifying agent are heated at least about 40 ℃.

In some embodiments of the inventive method, mixed step comprises and stirring the mixture that in other embodiments, whipping step forms continuous mixture.

In some embodiments of the inventive method, the step of mixture cures is comprised cools off mixture, in other embodiments, cooling step comprise with mixture be cooled to about 0 ℃ to about 3 ℃ temperature, or cure step mixes mixture during also being included in cooling step, or mixture is with the speed cooling of 1 ℃/min to about 20 ℃/min.

In some embodiments of the inventive method, cure step comprises nitrogen is imported in the mixture, and can comprise the nitrogen air-blowing through mixture.

Method of the present invention can comprise also at least a other composition is added in the mixture that described composition comprises water-soluble liquid (water-soluble liquid), comprises water.Can be by the amount interpolation water-soluble liquid of about 1wt.% to about 10wt.%.

Composition also can comprise at least a other composition (additional ingredient), and described composition comprises water-soluble liquid, comprises water.The amount of water-soluble liquid can be that about 1wt.% is to about 10wt.%.

Other composition also can be antioxidant, spices, flavoring agent, sweetener, pigment, vitamin, mineral matter, preceding biologic artifact (pre-biotic compound), probio compound (pro-bioticcompound), therapeutic component, drug ingedient, functional food composition (functional foodingredients), processing composition (processing ingredient), and their combination.

In some embodiments, other composition is the salt of ascorbic acid or ascorbic acid, in some embodiments, adds described composition with about 0.5wt.% to the amount of about 5wt.%.

In some embodiments, other composition is an antioxidant, in some embodiments, other composition is ascorbyl palmitate (ascorbyl palmitate), tocopherol, citric acid, ascorbic acid, tertiary butylated hydroquinone, Rosmarinus officinalis extract, lecithin or its mixture.

In some embodiments, the OSI value of solid-state fat composition be at least about 20, at least about 40 or at least about 60.

In some embodiments of the inventive method, solid-state fat composition is selected from food, nutriment and pharmaceutical products.

In some embodiments of the inventive method, method also comprises solid-state fat composition is added in the product that is selected from food, nutriment and the pharmaceutical products.

The present invention also provides and comprises that the microbial oil of winterization (unwinterized microbial oil) and about 0.01wt.% be not to the fat composition of the emulsifying agent of about 2.0wt.%, described microbial oil comprises the LC-PUFA and about 20wt.% saturated fat to about 60wt.% of about 5wt.% to about 70wt.%, wherein said composition comprises the water that is less than about 10wt.%, and wherein said composition is a solid-state composition in room temperature.

In another embodiment, the invention provides the method for preparing the oil product that is used to consume, described method comprises: extract the oil-containing fraction from microbial biomass, wherein said oil-containing fraction comprises at least a LC-PUFA and is enough to the saturated fatty acid of the described oil-containing fraction of visual impact at least; Handle described oil-containing fraction by vacuum evaporation, comprise the oil product of at least a LC-PUFA with preparation, wherein said oil product does not experience the winterization step.The present invention also provides the oil product by described method preparation.

The present invention also is provided for the microbial oil product that consumes, described oil product prepares by following steps: extract oil-containing fraction (oil-containing fraction) from microbial biomass, wherein said oil-containing fraction comprises at least a LC-PUFA and is enough to the saturated fatty acid of the described oil-containing fraction of visual impact at least; Handle described fraction by the method for vacuum evaporation, wherein said oil product does not experience the winterization step.

In some embodiments of the inventive method, oil product does not experience alkali refining process.In other embodiments, oil product does not experience the cold filtration process, and in other embodiments, oil product does not experience bleaching process.

In some embodiments of the inventive method, the oil-containing fraction can comprise that carbon chain lengths is at least 20, at least 22, has a LC-PUFA of at least three two keys or at least four two keys.In some embodiments, LC-PUFA can comprise DHA, clupanodonic acid, arachidonic acid or eicosapentaenoic acid.

In some embodiments of the inventive method, the step of handling the oil-containing fraction comprises the precipitation thinner.In other embodiments, the precipitation thinner can comprise, at high temperature vacuum condition is put on the oil-containing fraction of being extracted, described high temperature including, but not limited to about 50 ℃ to about 70 ℃ temperature.The precipitation thinner also can comprise, to put on the oil-containing fraction of being extracted greater than the vacuum of about 100mmHg vacuum, the oil-containing fraction of being extracted will be put on greater than the vacuum of about 70mmHg vacuum, or the oil-containing fraction of being extracted will be put on greater than the vacuum of about 50mmHg vacuum.

In some embodiments of the inventive method, the step of handling the oil-containing fraction comprises the precipitation thinner.In other embodiments, the precipitation thinner comprises, at high temperature vacuum condition is put on the oil-containing fraction of being extracted, and utilizes steam to spray the oil-containing fraction of being extracted simultaneously.In one aspect, high temperature is about 190 ℃ to about 220 ℃.In this embodiment, the precipitation thinner can comprise, to put on the oil-containing fraction of being extracted greater than the vacuum of about 25mmHg vacuum, the oil-containing fraction of being extracted will be put on greater than the vacuum of about 12mmHg vacuum, or the oil-containing fraction of being extracted will be put on greater than the vacuum of about 6mmHg vacuum.

In some embodiments of the inventive method, oil product experienced blanching step before or after treatment step.In other embodiments, method of the present invention also comprises oil is separated into olein (olein) fraction and stearin (stearin) fraction.In other embodiments, oil product is used for consuming for the people.

In some embodiments, oil product does not experience alkali refining process, cold filtration process or bleaching process.

In some embodiments, microbial biomass comes from and is selected from following microorganism: the microorganism that the microorganism that the microorganism that the microorganism that the microorganism that the microorganism that the microorganism that the microorganism of genus thraustochytrium, schizochytrium limacinum belong to, Althornia belong to, Aplanochytrium belong to, Japonochytrium belong to, Labyrinthula belong to, Labyrinthuloide belong to, Crypthecodinium belong to, and their mixture.In other embodiments, microbial biomass comes from and is selected from following microorganism: the microorganism that the microorganism that schizochytrium limacinum belongs to, Crypthecodinium belong to, and their mixture.

In some embodiments, oil product has the free fatty acid content less than about 0.5wt.%, in other embodiments, has the free fatty acid content less than about 0.3wt.%.

In some embodiments, oil product has the phosphorus value (phosphorousvalue) less than about 10ppm, in other embodiments, has the phosphorus value less than about 5ppm.

In some embodiments, oil product has the peroxide number (peroxidevalue) less than about 2meq/kg, in other embodiments, has the peroxide number less than about 1meq/kg.

In some embodiments, oil product has the anisidine value (anisidine value) less than about 5, in other embodiments, has the anisidine value less than about 3.

In some embodiments, oil product has the soap content (soap content) less than about 5wt.%, in other embodiments, has the soap content less than about 2.5wt.%.

In some embodiments, oil product has the Fe concentration less than about 1ppm, in other embodiments, has the Fe concentration of about 0.5ppm.

In some embodiments, oil product has the Pb concentration less than about 1ppm, in other embodiments, has the Pb concentration of about 0.2ppm.

In some embodiments, oil product has the Hg concentration less than about 0.1ppm, in other embodiments, has the Hg concentration of about 0.04ppm.

In some embodiments, oil product has the Ni concentration less than about 0.1ppm, and in other embodiments, oil product has the Ni concentration of about 0.01ppm.

In some embodiments, oil product has the Cu concentration less than about 1ppm, in other embodiments, has the Cu concentration of about 0.2ppm.

The present invention also provides the nutriment that comprises the microbial oil product, comprises the pharmaceutical products of microbial oil product and comprise the microbial oil product and the food of food or liquid composition.In some embodiments, pharmaceutical products also comprises pharmaceutically acceptable excipient.In other embodiments, pharmaceutical products also comprises and is selected from following medical active agent: the medicine of inhibin, antihypertensive, antidiabetic, anti-dull-witted medicine, antidepressants, antiadipositas drug, anorectic and raising memory and/or cognitive function.

In some embodiments, food is selected from dough (dough), batter (batter), bake food, food liquid, semisolid food, be pressed into the food (food bar) of piece, through pretreated meat (processedmeat), ice cream, frozen confectionery, the ice yoghourt, China's husband's compound (waffle mix), the salad flavoring, for egg compound (replacement egg mix), saline taste snacks (salted snack), special snacks (specialty snack), dried fruit snacks (dried fruit snack), meat flavour snacks (meat snack), pork rind, be pressed into the health food (health food bar) of piece, rice/corn-dodger (rice/corn cake) and confectionery snacks (confectionary snack).

In some embodiments, the microbial oil product is used for consuming for the people.

The present invention also is provided for the microbial oil product that consumes, and described oil product prepares by the method that may further comprise the steps: extract the oil-containing fraction that comprises at least a LC-PUFA from microbial biomass; Handle described fraction by the method for vacuum evaporation, wherein said oil product does not experience winterization step, alkali refining process, cold filtration process or bleaching process, and wherein said oil product has and is selected from following feature: free fatty acid content is less than about 0.5wt.%, the phosphorus value is less than about 10ppm, peroxide number is less than about 2meq/kg, anisidine value is less than about 5, soap content is less than about 5wt.%, Fe concentration is less than about 1ppm, Pb concentration is less than about 1ppm, Hg concentration is less than about 0.1ppm, Ni concentration less than about 0.1ppm and Cu concentration less than about 1ppm.

The present invention also provides the food that comprises microbial oil product and food or liquid composition, comprise the nutriment of microbial oil product and comprise the pharmaceutical products of microbial oil product.

The present invention also provides the method for preparing the oil product that is used to consume, and described method comprises: extract the oil-containing fraction from microbial biomass, wherein said oil-containing fraction comprises at least a LC-PUFA; Handle described oil-containing fraction by vacuum evaporation, comprise the oil product of at least a LC-PUFA with preparation, wherein said oil product does not experience alkali refining process.The present invention also provides the oil product of preparation by this method.

In some embodiments, microorganism is the microorganism of Mortierella (Mortierella).

In some embodiments, the oil-containing fraction comprises arachidonic acid.

The present invention also provides the oil product of mixing, described oil product comprises by the oil product of method 1 preparation and passes through the oil product that method 2 prepares, described method 1 comprises: extract the oil-containing fraction from microbial biomass, wherein said oil-containing fraction comprises at least a LC-PUFA and is enough to the saturated fatty acid of the described oil-containing fraction of visual impact at least; Handle described oil-containing fraction by vacuum evaporation, comprise the oil product of at least a LC-PUFA with preparation, wherein said oil product does not experience the winterization step; Described method 2 comprises: extract the oil-containing fraction from microbial biomass, wherein said oil-containing fraction comprises at least a LC-PUFA; Handle described oil-containing fraction by vacuum evaporation, comprise the oil product of at least a LC-PUFA with preparation, wherein said oil product does not experience alkali refining process.

In some embodiments, the microbial biomass of preparation method 1 oil product comes from and is selected from following microorganism: the microorganism that the microorganism that the microorganism of the microorganism that the microorganism that the microorganism that the microorganism that the microorganism of genus thraustochytrium, schizochytrium limacinum belong to, Althornia belong to, Aplanochytrium belong to, Japonochytrium belong to, Labyrinthula, genus, Labyrinthuloides belong to, Crypthecodinium belong to, and their mixture.In other embodiments, microorganism is selected from the microorganism of schizochytrium limacinum genus, the microorganism that Crypthecodinium belongs to, and their mixture.

In another embodiment of the oil product that mixes, the microbial biomass of preparation method's 2 oil products comes from the microorganism of Mortierella.

In another embodiment, the oil product of mixing comprises DHA and arachidonic acid.

On the one hand, in any one of embodiment of the present invention, the oil product by technology of the present invention or method preparation is a solid at 20 ℃.

Description of drawings

Fig. 1 shows multiple alternative embodiment of the present invention, and described embodiment prepares the oil of the PUFA of comprising of the present invention.

Fig. 2 shows multiple alternative embodiment of the present invention, and described embodiment prepares the oil of the PUFA of comprising of the present invention.

Fig. 3 shows the comparison of the oxidative stability index of following material, and described material is: solid-state fat composition, contain interpolation ascorbic acid solid-state fat composition and contain the ascorbic acid of interpolation and the solid-state fat composition of folic acid.

The specific embodiment

Instruct as the present invention, food compositions, nutrient composition and pharmaceutical products composition and prepare their method, make nutrient (LC-PUFAs especially, especially absorption ω-3 and ω-6 LC-PUFAs) increases, and described nutrient makes those objects that consume these products benefit that secures good health.The present invention partly relates to the oil product that contains PUFA with the high-quality of minimum processing procedure preparation, and described oil product has improved functional, has improved stability, and is suitable for very large-scale application, and these application comprise natural and/or organic market part.A kind of particularly preferred purposes of these oil products is the preparations that are used for solid-state fat composition, described composition comprises LC-PUFAs, and can in nutriment, food and/or pharmaceutical products (medicine and/or treatment), use, or as nutriment, food and/or pharmaceutical products (medicine and/or treat).The oil of preparation product of the present invention is the microbial oil that comprises LC-PUFAs that comes from microbial biomass.

First embodiment of the present invention is, prepares the method for the microbial oil of minimum processing procedure, and described microbial oil is the oil product that high-quality contains PUFA.This method comprises, extracts the oil-containing fraction that comprises at least a LC-PUFA from microbial biomass, with the preparation microbial oil.Microbe-derived and be used to that the method for growth of microorganism is well known in the art (Industrial Microbiology andBiotechnology, 2 ^NdEdition, 1999, American Society for Microbiology), described microorganism comprises nutrient and/or the LC-PUFAs that will reclaim in microbial oil.Preferably, in fermentation tank, in fermentation medium, cultivate microorganism.Method and composition of the present invention is applicable to any industrial microorganism that can produce LC-PUFA.

Microbe-derivedly comprise following microorganism: for example algae, bacterium, fungi (comprising yeast) and/or protist.Preferred organism comprises and is selected from those following organisms: chrysophyceae (for example microorganism of pipe hair living nature (Stramenopiles)), green alga, diatom (diatoms), dinoflagellate (dinoflagellates) (Dinophyceae (Dinophyceae) purpose microorganism for example, comprise the member that Crypthecodinium belongs to, for example to latent dinoflagellate (Crypthecodinium cohnii)), the fungi of yeast and mucor (Mucor) and Mortierella (including, but not limited to Mortierella alpina (Mortierella alpine) and Mortierella sect.schmuckeri).The member of pipe hair living nature micropopulation comprises microalgae and algae sample microorganism, comprises following microorganism: Hamatores, Proteromonads, Opalines, Develpayella, Diplophrys, Labrinthulids, thraustochytriale (Thraustochytrids), Biosecids, oomycetes (Oomycetes), Hypochytridiomycetes, Commation, Reticulosphaera, Pelagomonas, Pelagococcus, Ollicola, Aureococcus, Parmales, diatom, xanthophyta (Xanthophytes), Phaeophytes (brown alga), yellowish green algae (Eustigmatophytes), green whip algae (Raphidophytes), Synurids, Axodines (comprises Rhizochromulinaales, Pedinellales, Dictyochales), Chrysomeridales, Sarcinochrysidales, water tree algae (Hydrurales), live in seclusion chrysophyceae (Hibberdiales) and coloured gold algae (Chromulinales).Thraustochytriale comprises that (kind comprises aggregatum to the schizochytrium limacinum genus, limnaceum, mangrovei, minutum, octosporum), (kind comprises arudimentale to genus thraustochytrium, aureum, benthicola, globosum, kinnei, motivum, multirudimentale, pachydermum, proliferum, roseum, striatum), (kind comprises amoeboidea to the Ulkenia genus, kerguelensis, minuta, profunda, radiate, sailens, sarkariana, schizochytrops, visurgensis, yorkensis), (kind comprises haliotidis to the Aplanochytrium genus, kerguelensis, profunda, stocchinoi), Japonochytrium belongs to (kind comprises marinum), (kind comprises marisalba for Althornia genus (kind comprises crouchii) and Elina genus, sinorifica).Labrinthulids comprises that (kind comprises algeriensis to the Labyrinthula genus, coenocystis, chattonii, macrocystis, macrocystis atlantica, macrocystis macrocystis, marina, minuta, roscoffensis, valkanovii, vitellina, vitellina pacifica, vitellina vitellina, zopfi), Labyrinthomyxa belongs to (kind comprises marina), (kind comprises haliotidis to the Labyrinthuloides genus, yorkensis), Diplophrys belongs to (kind comprises archeri), Pyrrhosorus belongs to ^*(kind comprises marinus), Sorodiplophrys belong to ^*(kind comprises stercorea), Chlamydomyxa belong to ^*(kind comprises labyrinthuloides, montana).( ^*=current definite taxology ownership to these genus does not have consensus).Although can use method of the present invention to prepare the nutrient (these nutrients can produce) of various ways in a variety of microorganisms, but reason for succinct, convenient and example, following microbial growth method will be discussed: can generate the microorganism of the lipid that comprises ω-3 and/or omega 6 polyunsaturated fatty acid, particularly can generate the microorganism of DHA, DPA n-3, DPA n-6, EPA or ARA in detailed description of the present invention.Other preferred microorganism is an algae, the thraustochytriale of thraustochytriales (Thraustochytriales) for example, comprise that genus thraustochytrium (comprising Ulkenia) and schizochytrium limacinum belong to, and be included in the U.S.Patent Nos.5 that transfers Barclay jointly, 340,594 and 5,340, the thraustochytriales that discloses in 742, at this with its complete being incorporated herein by reference.More preferably, No. 20892, microorganism be selected from the microorganism with following microbial identification feature: ATCC No. 20888, No. 20890, No. 20889, ATCC, ATCC, No. 20891, ATCC and ATCC.Whether although be that the genus that is independent of genus thraustochytrium exists some differences with regard to Ulkenia between the expert, for purposes of this application, genus thraustochytrium comprises Ulkenia.Other is the bacterial strain (for example comprising the microorganism with ATCC 42430 identification marks) of the bacterial strain of Mortierella sect.schmuckeri (for example comprising the microorganism with ATCC 74371 identification marks) and Mortierella alpina preferably.Other comprises the microorganism of the identification mark with following microorganism: ATCC Nos.30021,30334-30348,30541-30543,30555-30557,30571,30572,30772-30775,30812,40750,50050-50060 and 50297-50300 preferably to the bacterial strain of latent dinoflagellate.Other preferably comes from the mentioned microorganism any one mutant strain, and their mixture.Oiliness microorganism (oleaginous microorganism) also is preferred.Use as the application, " oiliness microorganism " is defined as the microorganism more than 20% that to accumulate its cell weight with lipids form.The microorganism that can produce the genetic modification of LC-PUFAs also is suitable for the present invention.These microorganisms can comprise and carry out the microorganism that genetic modification can produce LC-PUFA naturally, and can not produce LC-PUFA naturally but carried out genetic engineering to produce the microorganism of LC-PUFA.

Can obtain suitable organism from multiple obtainable source, comprise by collecting from natural environment.American type culture collection (ATCC) has been listed the obtainable microbial strains of above identifying of the multiple public at present.Use as the application, the organism of microorganism or any particular type comprises wild strain, mutant or recombinant type arbitrarily.The growth conditions of cultivating these organisms is well known in the art, and disclosed the growth conditions that is suitable in these organisms at least some in following patent: for example United States Patent (USP) 5,130, and 242, United States Patent (USP) 5,407,957, United States Patent (USP) 5,397, and 591, United States Patent (USP) 5,492,938, United States Patent (USP) 5,711,983, United States Patent (USP) 5,882, and 703, United States Patent (USP) 6,245,365 and United States Patent (USP) 6,607,900, at this with the complete introducing of all these patents, as a reference.

Can pass through the known any suitable means of those skilled in the art, the microbial oil that can use in the present invention from microbe-derived recovery.For example, can be by coming recovered oil with following solvent extraction, described solvent is chloroform, hexane, carrene, methyl alcohol etc. for example, or comes recovered oil by supercritical fluid extraction.Alternatively, for example can be used all January 19 calendar year 2001 and submit to and denomination of invention is the United States Patent (USP) 6 of " Solventless Extraction Process ", 750,048 and PCT patent application US01/01806 in the extractive technique described extract oil, with these two pieces of complete introducings of patent documentation, as a reference.Other extraction and/or purification technique are instructed in following patent documentation: PCT patent application PCT/IB01/00841 that submit to April 12 calendar year 2001, and denomination of invention is " Method for theFractionation of Oil and Polar Lipid-Containing Native Raw Materials "; PCT patent application PCT/IB01/00963 that submit to April 12 calendar year 2001, denomination of invention is " Method for theFractionation of Oil and Polar Lipid-Containing Native Raw Materials UsingWater-Soluble Organic Solvent and Centrifugation "; The U.S. Provisional Patent Application 60/291 that submit to May 14 calendar year 2001,484, denomination of invention is " Production and Use of a PolarLipid-Rich Fraction Containing Stearidonic Acid and Gamma Linolenic Acidfrom Plant Seeds and Microbes "; The U.S. Provisional Patent Application 60/290 that submit to May 14 calendar year 2001,899, denomination of invention is " Production and Use of a Polar-Lipid FractionContaining Omega-3 and/or Omega-6 Highly Unsaturated Fatty Acids fromMicrobes, Genetically Modified Plant Seeds and Marine Organisms "; The United States Patent (USP) 6 that on February 17th, 2000 submitted to and on June 4th, 2002 was announced, 399,803, denomination of invention is " Process for Separating a Triglyceride Comprising a Docosahexaenoic AcidResidue from a Mixture of Triglycerides "; With the PCT patent application US01/01010 that submit to January 11 calendar year 2001, denomination of invention is " Process for Making an Enriched Mixture ofPolyunsaturated Fatty Acid Esters "; At this with the complete introducing of all these patent documentations, as a reference.Can carry out reduction vaporization to the oil that is extracted, with the sample of preparation enriched oil raw material.In following patent, disclosed and be used for living beings are carried out the enzyme processing to reclaim the method for lipid: the U.S. Provisional Patent Application 60/377 that on May 3rd, 2002 submitted to, 550, denomination of invention is " HIGH-QUALITYLIPIDS AND METHODS FOR PRODUCING BY ENZYMATIC LIBERATIONFROM BIOMASS "; PCT patent application PCT/US03/14177 that on May 5th, 2003 submitted to, denomination of invention is " HIGH-QUALITY LIPIDS AND METHODS FOR PRODUCINGBY ENZYMATIC LIBERATION FROM BIOMASS "; Ask 10/971,723 in the common unsettled United States Patent (USP) that on October 22nd, 2004 submitted to, denomination of invention is " HIGH-QUALITY LIPIDSAND METHODS FOR PRODUCING BY LIBERATION FROM BIOMASS "; European patent text 0 776 356 and United States Patent (USP) 5,928,696, denomination of invention is " Process forextracting native products which are not water-soluble from native substancemixtures by centrifugal force "; At this with the complete introducing of all these patent documentations, as a reference.

In preferred embodiments, rough microbial oil of the present invention (microbial crude oil) is the rough microbial oil of high-quality by method for preparing.For example compare with the fish oil of the raw oil that poor quality is provided usually, these oil of the present invention have significant advantage, for example this is because reclaim from the sashimi (raw fish) material and to be usually directed to boiling and hexane-extracted, and because described oil may comprise pollutant and/or other not desired components and/or the fatty acid profile do not expected.

The microorganism oil-containing fraction of extracting from microbial biomass that comprises at least a LC-PUFA as mentioned above comprises at least a LC-PUFA (promptly have 20 or the PUFAs of more a plurality of carbon).Preferred PUFAs of the present invention comprises C20, C22 or C24 ω-3 or ω-6PUFAs.Preferably, PUFA is long-chain PUFA (LC-PUFA), comprises C20 or C22 ω-3 or C20 or C22 omega 6 polyunsaturated fatty acid.LC-PUFA of the present invention comprises at least two two keys, preferably comprises three two keys, even more preferably comprises at least four two keys.Usually also will have 4 or the PUFAs of more a plurality of unsaturated carbon-carbon bonds be called high unsaturated fatty acid or HUFAs.Particularly, LC-PUFA can comprise that DHA (is at least about 10 of all fatty acids, about 20, about 30, about 40, about 50, about 60, about 70 or about 80% weight), clupanodonic acid n-3 (is at least about 10 of all fatty acids, about 20, about 30, about 40, about 50, about 60, about 70 or about 80% weight), clupanodonic acid n-6 (is at least about 10 of all fatty acids, about 20, about 30, about 40, about 50, about 60, about 70 or about 80% weight), arachidonic acid (is at least about 10 of all fatty acids, about 20, about 30, about 40, about 50, about 60, about 70 or about 80% weight) and/or eicosapentaenoic acid (be at least about 10 of all fatty acids, about 20, about 30, about 40, about 50, about 60, about 70 or about 80% weight).PUFAs can be any common form of finding in natural lipid, including, but not limited to the aliphatic acid of triacylglycerol, diacylglycerol, monoacylglycerol, phosphatide, free fatty, esterification, maybe can be the natural or synthetic derivative form (for example calcium salt of aliphatic acid, ethyl ester etc.) of these aliphatic acid.In preferred embodiments, the fraction that contains microbial oil comprises in described fraction at least about 70wt.%, at least about 80wt.%, at least about 90wt.% with at least about the PUFAs of the triglycerides form of 95wt.%.The term " LC-PUFA " that the present invention uses can refer to comprise single ω-3LC-PUFA (for example DHA) oil, comprise the oil of single ω-6LC-PUFA (for example ARA or DPA n-6), or comprise the oil of the mixture of two or more LC-PUFAs (for example DHA, DPA n-6, ARA and EPA).In preferred embodiments, product comprises LC-PUFA, and contains at least a other nutrient.

Except that using microbial biomass to be used for the oil extraction, the present invention also comprises use oily seed (oilseeds) as living beings, is used for extracting or reclaiming LC-PUFAs.Disclosed as the application, can process and handle these oil that extract from the oily seed living beings, with the preparation oil product.For example, oily seed comes from any higher plant, and especially expendable plant (consumable plant) comprises the farming plant, particularly the plant of using owing to its oil-containing.For example, these plants can comprise rape, soybean, coleseed, linseed, corn, safflower, sunflower and tobacco.Other preferred plant comprises known those plants that can produce as the compound of medical substance, aromatic substance, nutriment, functional food composition or beautifying active substance, or carries out genetic engineering to produce the plant of these composition/substance.Particularly preferred plant comprises and carried out genetic modification to produce the plant of LC-PUFAs, for example the gene of polyketide synthase systems imported plant wherein.For example, the United States Patent (USP) provisional application 60/671 of in following patent documentation, disclosed open text WO 2004/087879 A2, PCT of the method for these genes and Plant Transformation: PCT open text WO 02/083870 A2, PCT open text WO 2000/42195 A2, U.S. Patent Publication text US-2005-0100995-A1, submitting on April 15th, 2005,656 and U.S. Patent Publication text US-2005-0014231-A1, all these patents are all incorporated the application as a reference into.

By conventional method, for example by cleaning, shell and grinding and handle these seeds, with recovered oil.Then, can squeeze, preparing oil, or for example after rolling embryo, seed be contacted, with solvent to extract oil to seed.Suitable solvent can comprise organic solvent, the mixable solvent of water and water.Preferred solvent is a hexane.

In a plurality of embodiments of the present invention, another feature that comprises the oil product of PUFA is that they contain the saturated fatty acid that is enough to visual impact oil-containing fraction at least.The oil product of the multiple PUFA of comprising contains the saturated fatty acid of q.s, and described saturated fatty acid is in the form of the described oil of room temperature (promptly 20 ℃) energy visual impact, for example by produce muddiness and the form of the described oil of visual impact in oil.The product that some are such even owing to the existence of saturated fatty acid is paste, this for example is because they contain the saturated fatty acid that enough forms are triglycerides.Although in conventional method, these oil products are carried out winterization, to remove saturated fatty acid, the present invention recognizes, can not carry out under the situation of winterization, more goes through as following institute, prepares the product with commercial value from these oil products.

In a preferred embodiment of the invention, oil has and is particularly suitable for preparing the lipid profile that comprises the solid-state of LC PUFAs or semisolid composition.More specifically, these oil phases are rich in saturated compounds relatively to being rich in high unsaturated compound (for example 4,5 or more a plurality of unsaturated point), and/or be not rich in relatively single-, two-and three-saturated compounds.The feature of these compositions can be, with regard to saturation degree, has the compound that bifurcation distributes, and promptly the amount of saturated compounds is very high, and the amount of high unsaturated compound is also very high, and degree of unsaturation is low between the amount of the compound of centre.For example, these oil can contain and have 4 or the high unsaturated compound of more a plurality of unsaturated points, and its amount is for greater than about 20% weight, greater than about 25% weight, greater than about 30% weight, greater than about 35% weight, greater than about 40% weight, greater than about 45% weight or greater than about 50% weight.In other embodiments, these oil can contain and have 5 or the high unsaturated compound of more a plurality of unsaturated points, and its amount is for greater than about 20% weight, greater than about 25% weight, greater than about 30% weight, greater than about 35% weight, greater than about 40% weight, greater than about 45% weight or greater than about 50% weight.Available or extraly, these oil can contain saturated compounds, its amount is for greater than about 30% weight, greater than about 35% weight, greater than about 40% weight, greater than about 45% weight or greater than about 50% weight.Available or extraly, that these oil can contain is single-, two-or three-saturated compounds, its amount is for less than about 25% weight, less than about 20% weight, less than about 15% weight, less than about 10% weight or less than about 5% weight.

The inventive method that the high-quality for preparing minimum processing procedure contains the oil product of PUFA (comprising at least a LC-PUFA) also comprises, the oil-containing fraction of the extraction of preparation is as mentioned above handled.This extra processing comprises the method for vacuum evaporation, comprises the oil product of at least a LC-PUFA with preparation.

The precipitation thinner or the dry method of being undertaken by high vacuum evaporation are being known in the art, and comprise the oil that is extracted is applied vacuum condition, preferably apply vacuum condition at high temperature (for example about 50 ℃ to about 70 ℃).For example, can apply vacuum to described oil greater than about 100mmHg vacuum, greater than the vacuum of about 70mmHg vacuum or greater than the vacuum of about 50mmHg vacuum.For example, the application's's described " greater than vacuum of about 100mmHg vacuum " the meaning is stronger vacuum, for example 90mmHg or 80mmHg vacuum.Under these conditions, can remove any solvent, water or other component that boiling point in extraction oil is lower than described oil.

The method of deodorization is being known in the art, and comprises the oil that is extracted is applied vacuum condition, to remove any lower-molecular-weight component that may exist.Typically, remove these components by under high temperature and high vacuum, utilizing steam to spray.For example, usually described oil is applied vacuum greater than above those vacuums that thinner is mentioned with regard to precipitation.Particularly, vacuum can be greater than the vacuum of about 50mmHg vacuum, greater than the vacuum of about 25mmHg vacuum, greater than the vacuum of about 12mmHg vacuum or greater than the vacuum of about 6mmHg vacuum, usually can be extremely about 6mmHg vacuum of about 12mmHg vacuum, maybe can be that about 6mmHg vacuum is to about 1mmHg vacuum.This method also can be destroyed the multiple peroxide bond that may exist, and reduces or removes smell, and help to improve the stability of oil.In addition, under these conditions, can remove solvent, water or other component that boiling point in extraction oil is lower than described oil.Usually carry out deodorization at high temperature, for example about 190 ℃ to about 220 ℃ temperature.

The oil product that obtains from this method is the oil that high-quality contains PUFA, and described oil is used for for people and inhuman animals consuming, or described oil is suitable for for people and inhuman animals consuming.The organoleptic attribute of described oil makes that the consumption of described product is acceptable for people and inhuman animal.Particularly, described oil product can have low free-fat acid concentration, phosphorus value, peroxide number, anisidine value, soap content and heavy metal value.The preparation method of this oil of the present invention makes microbial oil reach the needed downstream processes amount of commercial receptive phase and minimizes.Concrete improvement comprises have been eliminated solvent winterization step, eliminated alkali refining process, eliminated the cold filtration process and may eliminate bleaching process.In addition, available deodorising process replaces the high vacuum evaporation process.Said method is described by keeping enough saturated compounds preventing composition in room temperature (promptly about 20 ℃) liquefy, thereby makes preparation solid-state or the semisolid product become easy.The present invention can be from rough microbial oil with extra high yield (95-100%) preparation edible oil, and described oil is suitable for natural and/or organic market part.

In a plurality of embodiments, oil product of the present invention, for example the oil for preparing under the situation that does not experience one or more conventional process steps has low free-fat acid concentration, and described conventional process step has solvent winterization, alkali refining process, cold filtration process and bleaching process.The mensuration of the free-fat acid concentration of oil is well-known in the art.More specifically, oil of the present invention can have less than about 0.5wt.%, less than about 0.1wt.% or less than the free fatty acid content of about 0.05wt.%.

In a plurality of embodiments, oil product of the present invention, for example the oil for preparing under the situation that does not experience one or more conventional process steps has low phosphorus value, and described conventional process step has solvent winterization, alkali refining process, cold filtration process and bleaching process.The mensuration of the phosphorus value of oil is well-known in the art.More specifically, oil of the present invention can have following phosphorus value, and described phosphorus value is less than about 10ppm, less than about 5ppm or for about 0ppm.

In a plurality of embodiments, oil product of the present invention, for example the oil for preparing under the situation that does not experience one or more conventional process steps has low peroxide number, and described conventional process step has solvent winterization, alkali refining process, cold filtration process and bleaching process.The mensuration of the peroxide number of oil is well-known in the art.More specifically, oil of the present invention can have following peroxide number, and described peroxide number is less than about 2meq/kg, less than about 1meq/kg or for about 0meq/kg.

In a plurality of embodiments, oil product of the present invention, for example the oil for preparing under the situation that does not experience one or more conventional process steps has low anisidine value, and described conventional process step has solvent winterization, alkali refining process, cold filtration process and bleaching process.The mensuration of the anisidine value of oil is well-known in the art.More specifically, oil of the present invention can have following anisidine value, described anisidine value less than about 5, less than about 3, less than about 2, less than about 1, less than about 0.5, less than about 0.3, less than about 0.1 or be lower than detectability.

In a plurality of embodiments, oil product of the present invention, for example the oil for preparing under the situation that does not experience one or more conventional process steps has low soap content, and described conventional process step has solvent winterization, alkali refining process, cold filtration process and bleaching process.The mensuration of the soap content of oil is well-known in the art.More specifically, oil of the present invention can have following soap content, and described soap content is less than about 5wt.%, less than about 2.5wt.% or be 0wt.%.

In a plurality of embodiments, oil product of the present invention, for example the oil for preparing under the situation that does not experience one or more conventional process steps has low heavy metal value, and described conventional process step has solvent winterization, alkali refining process, cold filtration process and bleaching process.The mensuration of the heavy metal value of oil is well-known in the art.More specifically, oil of the present invention can have less than about 1ppm, less than about 0.5ppm or be preferably about the Fe concentration of 0ppm; Can have less than about 1ppm, less than about 0.2ppm or be preferably about the Pb concentration of 0ppm; Can have less than about 0.1ppm, less than about 0.04ppm or be preferably about the Hg concentration of 0ppm; Can have less than about 0.1ppm, less than about 0.01ppm or be preferably about the Ni concentration of 0ppm; Can have less than about 1ppm, less than about 0.2ppm or be preferably about the Cu concentration of 0ppm.

The inventive method that the high-quality for preparing minimum processing procedure contains the oil product of PUFA (it has at least a LC-PUFA) can be chosen wantonly and comprise, the step of before or after deodorization step or high vacuum separating step described oil product being bleached is although blanching step carries out before being more typically in the deodorization step.Bleaching to oil is well-known in the art, and available conventional method is bleached.Particularly, for example silica gel (silica) adsorbent (for example Trysil 600 (GraceChemicals)) and the bleaching clay (bleaching clay) that is used for removing residual soap can be incorporated into oil, then it be filtered out.Usually, silica gel absorber adds after bleaching clay.

The inventive method that the preparation high-quality contains the oil product (it has at least a LC-PUFA) of PUFA can comprise that preparation comprises the fluid oil fraction of LC PUFA and comprises the method for the solid-state fat products of LC PUFA.This method may further comprise the steps: disclosed as the application, high-quality rough microbial oil is separated into oil product and relevant solid-state fat products.Can prepare these raw oil products by the following method, described method is for extracting the oil-containing fraction that comprises at least a LC-PUFA and saturated fatty acid from microbial biomass.Can handle the oil-containing fraction by winterization, cold filtration, vacuum evaporation and/or other means, comprise the fluid oil product and the solid product that comprises at least a LC-PUFA of at least a LC-PUFA with preparation.These other means can comprise filtration, so that the fluid oil fraction is separated with solids fraction.

Can reclaim solid-state fraction component (may comprise adsorbent) by solid/liquid separation technique.Can solid-state fatty material be melted by adding heat-adsorbent and solid-state fatty material, any adsorbent is separated with solid-state fraction.Then, can adsorbent be separated with the solid of thawing, can the solid of thawing be solidified again by cooling then for example by filtering.

The solid-state fraction that reclaims can comprise the LC PUFA of high-load.In preferred embodiments, solid-state fraction can comprise at least about 20%, at least about 25% or at least about LC PUFA, the especially DHA of 30% weight.Every kind in the oil of clarification and the solid all can be used as for example food or food additive.

Oil product prepared in accordance with the present invention can be solid-state or semisolid.It is that solid-state or semi-solid those materials reach at room temperature those materials for liquid state that the employed term of the application " oil " can be included in room temperature.

The inventive method that the high-quality for preparing minimum processing procedure contains the oil product of PUFA (it has at least a LC-PUFA) can be chosen wantonly and may further comprise the steps: after deodorization step or high-vacuum fractionation step, oil content is heated up in a steamer (fractionate) and become olein fraction and stearin fraction.Oil is fractionated into olein fraction and stearin fraction, can be with this step application in the oil of any raw oil, bleaching or the oil of deodorization, with the olein fraction of preparation clarification and hard stearin fraction.Olein can use in different food applications owing to the different of its physical property with stearin.In conventional method, stearin is the byproduct of oil water mixture winterization and cold filtration, and its processing is caused～30% loss.Fractionating step can prepare the stearin fraction that is suitable for selling.Below in embodiment 5, show the embodiment of this fractionation.

With reference to figure 1, explain multiple alternative embodiment of the present invention.The solvent that utilization is used to extract raw oil is to initiation material, living beings for example, and for example spray-dired living beings are handled.These raw oils can comprise long-chain polyunsaturated fatty acid.Can carry out high vacuum evaporation to raw oil, this extraction solvent, water and other component that the raw oil mid-boiling point can be lower than desired oil ingredient is removed.Alternatively, the blanching step that can choose wantonly raw oil, thus for example remove carotenoid.Then, spray with steam, the raw oil of optional treatment is carried out deodorization by under high temperature and high vacuum, making oil.Then, the final oil product for preparing by high vacuum evaporation or deodorization can be chosen wantonly by the following method and handle: it is fractionated into olein fraction and stearin fraction.

With reference to figure 2, explain multiple alternative embodiment of the present invention by flow chart.Method must may further comprise the steps in its most basic form: begin with the pasteurized fermented liquid that comprises microbial biomass.Zymotic fluid is carried out preliminary treatment,, for example realize by enzyme processing or mechanical disruption so that oil disengages from cell owing to cytolysis.Then, pretreated zymotic fluid is carried out extraction step, with the preparation microbial oil.This method comprises deodorization step as described in the present application then at least.In a kind of alternative method, method comprises blanching step, before the deodorization step, the microbial oil that is extracted is bleached whereby.In other alternative embodiment, can carry out winterization step (being cold filtration) to the microbial oil that is extracted before the blanching step and/or between blanching step and deodorization step.

The method and the resulting product that prepare the oil (minimally processed oils) of minimum processing procedure of the present invention have many significant advantages.The conventional method that comprises the oil product of PUFA with preparation is compared, and the present invention has lower cost, has reduced the processing procedure necessary condition, has increased output, has improved the treatment step security, and has eliminated refuse/by-product stream.And method of the present invention is consistent with natural and/or organic market part.The conventional method of oil processing is used the downstream processes of all aspects usually, comprises chemical refining.Also do not expand physics process for purification (method that does not promptly relate to alkali refining) to fish oil and similarly comprise the oil of PUFA, this may be owing to have known difficulty in these oily processing procedures.And because the restriction of aroma and flavor, many known physical treatment methods or refining inadequately product are restricted.Surprisingly, method of the present invention uses physical method and minimal steps to prepare taste oil preferably.

Describe more fully as following institute, can in numerous food and food applications, use high-quality of the present invention to contain the oil product of PUFA.Described oil product can be used as nutriment, meal product, medicine or pharmaceutical products and directly consumes for the people.In addition, described oil product can be combined with known arbitrarily human foods or liquid for people's consumption, to improve nutrition.Described oil product also can be directly as feed or as the tonic of animal feed and feed to animal.So, arbitrarily based on the food of animal when the confession people consumes, can have the quality of raising.

In one embodiment, can use oil product of the present invention, infant formula is replenished.Infant formula for example can be supplemented with the refining oil of physics that is derived from the microorganism (for example Mortierella alpina or Mortierella sect.schmuckeri) that can produce ARA, the refining oil of described physics replenishes separately or for example (for example microbial oil comprises DHA-S for fish oil or other are rich in the oil of DHA with other oil ^TMAnd DHA-T ^TMOil (Martek Biosciences, Columbia, MD)) replenish together.These refining oil of physics that comprise ARA should not accepted chemical refining.Alternatively, infant formula for example can be supplemented with the oil of the minimum processing procedure that is derived from the microorganism that can produce DHA (for example to latent dinoflagellate), the oil of described minimum processing procedure replenish separately or with other oil that is rich in ARA, comprise ARASCO ^(Martek Biosciences, Columbia MD) replenish together.In another embodiment, infant formula can be supplemented with the oil multiple of the present invention that is derived from one or more sources, for example comprise the minimum processing procedure of DHA (for example come to latent dinoflagellate) oil and comprise the refining oil of physics of ARA (for example coming from Mortierella alpina).

In other embodiments, can be with oil product combination of the present invention, with the preparation mixture.For example, can be with the oil and the refining oil mixing of the physics that comes from Mortierella alpina that is derived from the minimum processing procedure of latent dinoflagellate, to be used for supplementing infant formula.Can use oil of the present invention, with multiple different ARA and DHA ratio, preparation comprises the oil of ARA and comprises the mixture of the oil of DHA.These mixtures can comprise about 1: 1 to about 2: 1 ARA: the DHA ratio.More specifically, can prepare ARA: the DHA ratio is the mixture of about 1: 1,1.25: 1,1.5: 1,1.75: 1 or 2: 1.

In particularly preferred embodiments, can use high-quality of the present invention to contain the oil product of PUFA, as the initiation material of the solid-state fat composition of following detailed description.Yet the purposes that it will be appreciated that the oil product of minimum processing procedure of the present invention is not limited to the initiation material as the described solid-state fat composition of the application.

The present inventor has surprisingly been found that, in the preferred embodiment of the solid-state fat composition of the present invention, can use the not winterization form of the oil that is rich in LC-PUFA, comprise the not oil that comprises DHA that comes from microorganism (DHA oil) of winterization, as the initiation material of the solid-state fat composition of the present invention.Thus, prepare these method for compositions and can no longer need following steps: oil is carried out hydrogenation, with these oil with firmly or saturated fat or other thickening type material mix.Usually, be that liquid fish oil or microbial oil are prepared into initial raw oil with refining oil, then purification step (removing phosphatide and free fatty) and blanching step (removing pigment) are applied to described raw oil.Usually, then oil is carried out winterization, to remove saturated fat.

The present inventor has surprisingly been found that, for example, the microbial oil of winterization does not promptly carry out the microbial oil of winterization step, provides the initiation material of the processing procedure that need not the prior art instruction, to form solid-state composition.In addition, can use the oily seed oil of aforesaid not winterization, as substituting of microbial oil as described below.Not bound by theory, the present inventor believes, the saturated fat that is present in the oil of winterization is not given more solid-state denseness (comparing with the fluid oil of winterization) to described oil.The method that the present invention prepares solid-state fat composition has also overcome the tendency that particle (because crystallization of triglycerides) appears in the oil of winterization not, the appearance of particle make these not the oil of winterization look like the thick fluid oil that contains particle.When room temperature is placed, separating does not appear in the oil of winterization, and products obtained therefrom looks like the thick fluid oil that wherein contains solid.The present invention can overcome this feature of nonwinterized oil.Method of the present invention prepares the uniform smooth product of outward appearance, and described product is when be stable (any separation not occurring) when room temperature is placed.Products obtained therefrom can have the denseness of shortening.

In another embodiment, the present invention includes the method for the solid-state fat composition of preparation.This method comprises that described oil comprises saturated fat and contains the microbial oil of at least a LC-PUFA with oil and the step of at least a emulsifier with the formation mixture.Then, make the gained mixture solidified, form solid-state fat composition.

" solid-state fat composition " refers to be solid-state or semi-solid composition in room temperature (promptly 20 ℃).The physicochemical property of fat and oil comprises its viscosity and melt temperature.Preferably, solid-state fat composition can have at least about 20 ℃, at least about 25 ℃ or at least about 30 ℃, be preferably melt temperature at least about 35 ℃.Melt temperature can change significantly, and this depends on the quantity of existing different chemical entity.Usually, and based on the fusing point of each triglycerides and comparing of expecting, the mixture of some triglycerides has lower fusing point.Compare with the melting range of each component of mixture, mixture also can have the melting range of broad.Compare with the triglycerides of similar aliphatic acid composition, monoglyceride and diglyceride have high melt point.In preferred embodiments, solid-state fat composition can be enough soft, to be applied on the food.Preferably, composition can be a thickness in room temperature, has sluggish flowing property, and/or compare the easier surface that adheres to of composition with the initiation material of preparing product.

Prepare the oil that uses in the solid-state fat composition method in the present invention and comprise the microbial oil that contains at least a LC-PUFA.Described in detail in the explanation as above oil in minimum processing procedure of the present invention, microbe-derived and be used to make the method for growth of microorganism to be well known in the art, described microorganism comprises nutrient and/or the LC-PUFAs that will reclaim in microbial oil.These microbe-derived and methods also are suitable for preparing the microbial oil as the solid-state fat composition initiation material of the present invention.In fact, the oil of aforesaid minimum processing procedure is the preferred initiation material of the solid-state fat composition of preparation.Yet, it will be appreciated that and can use multiple other microbial oil initiation material as described below, as the initiation material of the solid-state fat composition of the present invention.In an especially preferred embodiment, microbial oil is the oil according to the disclosed content preparation of following patent application, described patent application is PCT patent application PCT/IB01/00841, denomination of invention is " Method for the Fractionationof Oil and Polar Lipid-Containing Native Raw Materials ", submit publication number WO 01/76715 in April 12 calendar year 2001; PCT patent application PCT/IB01/00963, denomination of invention is " Method for the Fractionation of Oil and Polar Lipid-Containing Native RawMaterials Using Water-Soluble Organic Solvent and Centrifugation ", submit in April 12 calendar year 2001, publication number is WO 01/76385.The content description that this two PCT application discloses the microbial oil recovery method, described method can be called the Friolex method usually.

Microbial oil of the present invention comprises at least a LC-PUFA (promptly have 20 or the PUFAs of more a plurality of carbon).Preferred PUFAs of the present invention comprises C20, C22 or C24 ω-3 or ω-6PUFAs.Preferably, PUFA is long-chain PUFA (LC-PUFA), comprises C20 or C22 ω-3 or C20 or C22 omega 6 polyunsaturated fatty acid.LC-PUFA of the present invention comprises at least two two keys, preferably comprises three two keys, even more preferably comprises at least four two keys.Usually also will have 4 or the PUFAs of more a plurality of unsaturated carbon-carbon bonds be called high unsaturated fatty acid or HUFAs.Particularly, LC-PUFA can comprise that DHA (is at least about 10 of all fatty acids, about 20, about 30, about 40, about 50, about 60, about 70 or about 80% weight), clupanodonic acid n-3 (is at least about 10 of all fatty acids, about 20, about 30, about 40, about 50, about 60, about 70 or about 80% weight), clupanodonic acid n-6 (is at least about 10 of all fatty acids, about 20, about 30, about 40, about 50, about 60, about 70 or about 80% weight), arachidonic acid (is at least about 10 of all fatty acids, about 20, about 30, about 40, about 50, about 60, about 70 or about 80% weight) and/or eicosapentaenoic acid (be at least about 10 of all fatty acids, about 20, about 30, about 40, about 50, about 60, about 70 or about 80% weight).PUFAs can be any common form of finding in natural lipid, including, but not limited to the aliphatic acid of triacylglycerol, diacylglycerol, monoacylglycerol, phosphatide, free fatty, esterification, maybe can be the natural or synthetic derivative form (for example calcium salt of aliphatic acid, ethyl ester etc.) of these aliphatic acid.In preferred embodiments, microbial oil comprises in described oil at least about 70wt.%, at least about 80wt.%, at least about 90wt.% or at least about the PUFAs of the triglycerides form of 95wt.%.The term " LC-PUFA " that the present invention uses can refer to comprise single ω-3LC-PUFA (for example DHA) oil, comprise the oil of single ω-6LC-PUFA (for example ARA or DPA n-6), or comprise the oil of the mixture of two or more LC-PUFAs (for example DHA, DPA n-6, ARA and EPA).In preferred embodiments, product comprises LC-PUFA, and contains at least a other nutrient.

In a preferred embodiment of the invention, preparing the oil that uses in the solid-state fat composition method in the present invention can comprise at least about 5wt.%, at least about 10wt.%, at least about 15wt.%, at least about the LC-PUFA of 20wt.%, at least about 25wt.%, at least about 30wt.%, at least about the LC-PUFA of 35wt.%, at least about 40wt.%, at least about 45wt.% or at least about the LC-PUFA of 50wt.%.These embodiments also can have less than about 30wt.%, less than about 35wt.%, less than about 40wt.%, less than the LC-PUFA of about 45wt.%, less than about 50wt.%, less than about 55wt.%, less than about 60wt.%, less than the LC-PUFA of about 65wt.% or less than the LC-PUFA of about 70wt.%

Prepare the oil that uses in the solid-state fat composition method in the present invention, except that comprising the microbial oil that contains at least a LC-PUFA, also comprise saturated fat.Compare with the mixture of LC-PUFA or LC-PUFAs, saturated fat can have high melt point usually.This saturated fat external source can be added in the oil.The saturated fat that preferred external source is added comprises " hard fat (hard fat) ", oil, partially hydrogenated lard and the non-trans tropical oil (tropicaloil) of for example partially hydrogenated vegetable oil, hydrogenation fully.For example, can use palm oil, palm-kernel oil (palm kernel oil) and fraction (palm olein and palm kernel olein, palm stearines and palm kernel stearin) thereof.When composition comprises fatty that external source adds, can carry out winterization to LC-PUFA oil, also can not carry out winterization.Those skilled in the art can determine the preferred amounts of external source interpolation fat based on the hardness of initiation material and/or the coating denseness (spread consistency) of viscosity and the hardness of expecting and/or viscosity and/or expectation in composition.The fat that external source is added can be by about 20wt.% to extremely about 50wt.% or the extremely amount interpolation of about 45wt.% of about 35wt.% of about 60wt.%, about 30wt.%.

In preferred embodiments, saturated fat is not that external source is added, but natural being present in the microbial oil.For example, the microbial oil that comprises LC-PUFAs can be a untreated oil, and described untreated oil extracts by any means known in the art.In these oil, the amount of saturated fat can be about 20wt.% to about 60wt.%, about 30wt.% about 50wt.% or about 35wt.% about 45wt.% extremely extremely in the microbial oil.

In a preferred embodiment of the invention, microbial oil is (promptly unsegregated) of winterization not, therefore comprises saturated fat.Winterization refers to remove at low temperature the method for the deposit (being generally the solid-state saturated fat of high-melting-point) that occurs in the multiple oil (comprising vegetable oil), more typically relates to removing by filter a certain amount of crystalline material to prevent that liquid fraction is in refrigerated storage temperature (refrigerator temperature) appearance muddiness.These technology comprise two or more fractions that oil are separated into different melting points.Liquid fraction separately and solid-state fraction show very big different aspect physics and chemical property.Suitable technique is well known in the art, and generally includes following three steps: (i) fluid oil is cooled to supersaturation, forms nucleus, be used for crystallization; (ii) cooling is gradually constantly grown crystal; And liquid phase and crystal are separated.These technology for example comprise traditional winterization, detergent fractionation (detergent fractionation) and solvent winterization.The tradition winterization comprises dry classification crystallization (dry fractional crystallization), wherein the triglycerides that melting temperature is the highest during cooling, preferentially crystallization from the fat of clean (neat) liquid or thawing.The principle of dry fractionation (dry fractionation) method is based on, and under the condition of control, under the situation of not adding chemical substance, makes oil cooling but.By mechanical means, liquid phase is separated with solid phase.The principle of detergent fractionation is similar to dry fractionation, also is based under the condition of control, under the situation of not adding solvent, makes oil cooling but.Next, after adding the detergent aqueous solution,, liquid phase is separated with solid phase by centrifugal.Utilize solvent (being generally acetone) winterization to promote the formation of triglycerides crystal, this is because compare with the situation that does not have solvent, is having under the situation of solvent, and triglycerides forms more stable crystal usually at low temperature.In the auxiliary fractionation of solvent, can use polarity or non-polar solven, during filtering, to reduce the viscosity of system.Then, the gained fraction is desolvated by distilling to remove.Thereby the microbial oil of winterization is not for living through those microbial oils of winterization or separation process.

In other embodiment preferred, microbial oil be do not have hydrogenation or do not have partially hydrogenated.Hydrogenation is well known in the art, and comprises following method: existing under the situation of catalyst, the hydrogen chemistry is added to liquid fat.This method changes into singly-bound with in two keys of unrighted acid in the fat molecule at least some, improves the saturation degree of fat thus.The degree of hydrogenation promptly transforms the sum of two keys, the physics and the chemical property of decision hydrogenated fat.Partially hydrogenated oil keeps sizable degree of unsaturation usually in its aliphatic acid.Hydrogenation also makes some cis-double bonds change into anti-configuration, and wherein one or more two keys move to new position in fatty acid chain.Current research shows that trans-fatty acid can improve T-CHOL, and increases the heart disease risk, and its degree is identical with saturated fatty acid, does not therefore expect in diet.The present invention can need not to prepare solid-state or semisolid product under hydrogenation or the partially hydrogenated situation.Method of the present invention comprises mixes at least a emulsifying agent with the oil that comprises the microbial oil with at least a LC-PUFA.The preferred emulsifier that uses with the present invention comprises monoglyceride, diglyceride, monoglyceride/diglyceride combination, lecithin, dilactic acid monoglyceride-diglyceride, polyglycerol ester, sucrose fatty ester, stearyl dilactic acid sodium, stearyl dilactic acid calcium, and their combination.In preferred embodiments, emulsifying agent is monoglyceride/diglyceride combination.In preferred embodiments, emulsifying agent by with about 0.01% weight to about 2.0% weight, about 0.025% weight to about 1.0% weight or about 0.05% weight to the amount of about 0.2% weight be present in the mixture.Not bound by theory, emulsifying agent is considered to make in the mixture various components stable, to keep the composition of homogeneous.The shortage of stability can cause the separation of oil or separating of oil and water.Emulsifying agent also can provide the functional performance except that emulsification, comprises foam (aeration), makes starch and protein is compound, hydration, crystal modified, solubilising and dispersion.

Emulsifying agent is carried out with any conventional hybrid mode that is known in the art with the physical step that oil mixes.Component is mixed, realize mixing, thereby obtain the liquid solution of homogeneous.For example, may be essential be, microbial oil and/or emulsifying agent for example are heated at least about 40 ℃, be liquid fully thereby make component, miscible each other.In preferred embodiments, oil is the oil that is rich in LC-PUFA of winterization not, is heated at least about 40 ℃, so that all components of oil all dissolves.In preferred embodiments, emulsifying agent is the mixture of monoglyceride emulsifying agent and diglyceride emulsifying agent, and it in the vessel in heating of opening with oil content, is formed liquid.Then, by any known method, preferably, the oil that melts is in the same place with emulsifier, to form continuous mixture by stirring.

Method of the present invention also comprises the mixture solidified that makes oil and emulsifying agent, to form solid-state fat composition.For example, in the preferred embodiment on mixture is in room temperature, mixture can be cooled to room temperature.Alternatively, can on one's own initiative mixture be cooled to room temperature, or for example be cooled under the room temperature.For example, in order to solidify, can be with composition cools to about 0 ℃ to about 3 ℃.During cooling step, no matter initiatively still passive cooling of cooling all can mix or stirs mixture.So, can control, thereby realize cooling uniformly, and can not form the composition of layering cooling.Preferably, adjust these cooling conditions, so that make the crystal structure (being the mode that molecule self is orientated in solid phase) of fat reach the level of expectation, thus the product plasticity that obtains expecting, functional and stable.Usually, β ' crystal obtains even butyraceous denseness.Compare with β ' crystal, the β crystal normally more greatly, more coarse and be granular, therefore normally more do not expect.Therefore, in preferred embodiments, the control cooling procedure, thus make the triglycerides in the mixture form stable β ' crystal formation, the product that has even denseness with preparation.The cooling means that obtains these preferred crystal forms comprises by following speed cooling mixture, described speed be about 1 ℃/min to about 20 ℃/min or about 5 ℃/min to about 15 ℃/min, or be about 10 ℃/min.Not bound by theory, the present inventor believes, is suitable for the some emulsifier that uses with the present invention, for example monoglyceride and diglyceride, and the crystallization of triglycerides in influence and/or the control combination thing at least in part, thus obtain β ' crystal.Preferably, in solid-state fat composition, at least about 50wt.%, at least about 55wt.%, at least about 60wt.%, at least about 65wt.%, at least about 70wt.%, at least about 75wt.%, at least about 80wt.%, at least about 85wt.%, at least about 90wt.%, at least about 95wt.% or about 100wt.% fat and/or oil be β ' crystal configuration.

In other embodiments, the mixture solidified that makes oil and emulsifying agent can comprise importing nitrogen to form the step of solid-state fat composition, makes it through mixture.For example, the nitrogen air-blowing can be entered in the composition.Alternatively, can when emulsification, nitrogen be imported in the low temperature crystallization device.

The importing of nitrogen during curing can improve the oxidation stability of product, and can be by making the glossy product appearance of improving of outward appearance.

In preferred embodiments, solid-state fat composition of the present invention has the structure of homogeneous, therefore has uniform outward appearance and denseness.Another of these embodiments is characterized as, and preferably with regard to for a long time, composition is stable, can not occur separating when placing, and also can not lose the structure of its homogeneous.Thereby composition can not develop and uneven outward appearance or denseness when placing.In preferred embodiments, composition of the present invention can room temperature place at least about one day, at least about a week, at least about two weeks, at least about three weeks or at least about around, and the structure of its homogeneous can not appear separating or losing.

Composition of the present invention also can comprise multiple other functional components.For example, composition of the present invention also can comprise microencapsulation thing (microencapsulant), for example comprises protein, simple and complicated carbohydrate, solid and particle.Preferred microencapsulation thing comprises cell particle, Arabic gum, maltodextrin, hydrophobically modified starch, polysaccharide (comprising alginates, carboxymethyl cellulose and guar gum), hydrophobically modified polysaccharide (for example starch of octyl group replacement), protein (comprising whey isolate protein (wheyprotein isolate), soybean protein and casein sodium), and their combination.In addition, composition of the present invention can comprise surfactant, for example comprises anion surfactant, cationic surfactant, nonionic surfactant, amphoteric surfactant; The particle of water-insoluble emulsifying agent, fine dispersion and naturally occurring material.Anion surfactant comprises carboxylic acid, sulfuric ester, alkane sulfonic acid, alkylated aromatic sulfonic acid, various anionic hydrophilic group; Cationic surfactant comprises amine salt, ammonium compounds, other nitrogenous alkali, unazotized alkali; Nonionic surfactant comprises ehter bond, ester bond, amido link, various key (miscellaneous linkage), the multiple bond (multiple linkages) that combines with solubilizing group; Amphoteric surfactant comprises the various combinations of amino and carboxyl, amino and sulfuric ester, amino and alkane sulfonic acid, amino and aromatic hydrocarbons sulfonic acid, basic group and acidic-group; The water-insoluble emulsifying agent comprises ionic hydrophilic radical, nonionic hydrophilic radical; The particle of fine dispersion comprises the non-solubilising particle of any fine dispersion, comprises clay and carbon; Naturally occurring material comprises alginates, cellulose derivative, water-soluble glue, lipid and sterol, phosphatide, aliphatic acid, alcohol, protein, amino acid, detergent; And hydrophilic colloid.Other optional ingredients comprises thickener, and described thickener comprises polysaccharide.Thickener is the composition that is used to increase composition viscosity.In these embodiments, during blend step, add other functional components (or multiple functional components) usually.

In one embodiment, solid-state fat composition is a shortening.Shortening has the water or the aqueous components of interpolation seldom usually, or does not have the water or the aqueous components of any interpolation, and comprises the fat of high-load.Alternatively, solid-state fat composition can be following product, for example margarine, daubing product, mayonnaise or salad flavoring.By fat and/or oil are mixed with other composition, prepare these products, described other composition for example is water and/or dairy produce, suitable edible protein, salt, flavoring substance and coloring material and vitamin A and D.Margarine comprises at least 80% fat usually.Mayonnaise and salad flavoring are semisolid fatty food, and these two kinds of foods comprise respectively usually and are no less than 65% and 30% vegetable oil and dried whole-egg or yolk.Salt, sugar, spices, condiment, vinegar, lemon juice and other composition make these products perfect.

Therefore, the present invention also can comprise and has water-soluble liquid or water-soluble liquid is added in the mixture.Preferably, water-soluble liquid is a water, and by less than about 10wt.%, is about 1wt.% amount interpolation to about 6wt.% to about 10wt.%, about 2wt.% to about 8wt.%, about 4wt.%.The existence of water-soluble liquid allows to add one or more other water soluble ingredients.Water soluble ingredient all is suitable for the present invention arbitrarily.Preferred other composition comprises antioxidant, spices, flavoring agent, sweetener, pigment, vitamin, mineral matter, preceding biologic artifact, probio compound, therapeutic component, drug ingedient, functional food composition, processing composition, and their combination.

In particularly preferred embodiments, other composition is an antioxidant.Antioxidant is well known in the art, and can add in the arbitrfary point in the following process: prepare the process of microbial oil, the processing procedure or the processing procedure of the present invention of lipid by fermentation.Antioxidant can help to prevent the products obtained therefrom oxidation deterioration.The available suitable antioxidant of technical staff.Preferred anti-oxidants comprises ascorbyl palmitate, tocopherol, citric acid, ascorbic acid, tertiary butylated hydroquinone (TBHQ), Rosmarinus officinalis extract, lecithin, and their mixture.Antioxidant can be included in the product by the traditional amount in this area.Particularly preferred antioxidant comprises the salt of ascorbic acid or ascorbic acid.In preferred embodiments, when antioxidant was the salt of ascorbic acid or ascorbic acid, oxidant can be by about 5wt.% at the most, comprised that about 0.5wt.% is to about 5wt.%, about 1.5wt.% to about 5wt.% with the extremely amount interpolation of about 5wt.% of about 3wt.%.It should be noted that when adding water soluble antioxidant (for example ascorbic acid, citric acid or its salt), described antioxidant must add with water, thereby it is dispersed in the composition well.Have surprisingly been found that the increase level of the oxidation stability of product of the present invention is higher than the level of expecting with regard to the amount of used antioxidant, especially when antioxidant is the salt of ascorbic acid or ascorbic acid.For example, add ascorbic acid or its salt of about 5wt.%, make the OSI (oxidative stability index, oxidative stability index) of the present composition increase by three times.

The state of oxidation and the stability that comprise the composition of lipid can be measured by the several different methods that is known in the art, and can obtain from American Oil Chemist ' s Society and other source the description of multiple these technology.It is by measuring oxidative stability index (OSI) that the product oxidation stability is carried out quantitative a kind of method, for example, measure the amount of the volatile decomposition products (conductive species) of when sample being carried out the thermal decomposition operation, from sample, disengaging by using the Rancimat instrument.In preferred embodiments, composition of the present invention has at least about 10, at least about 20, at least about 30, at least about 40, at least about 50, and at least about 60 OSI value.

In preferred embodiments, product of the present invention (comprising that high-quality contains oil product and the solid-state fat composition of PUFA) is stored under the appropriate condition, so that oxidative degradation minimizes.The several different methods that realizes these storage requirements is well known in the art, and is suitable for using with the present invention, for example replaces surrounding air with inert gas atmosphere.Making oxidative degradation minimizing or minimized method for optimizing is that product is stored in nitrogen (N ₂) under the nitrogen and carbon dioxide atmosphere of atmosphere or mixing.Preferably, need packaged products under nitrogen, to pack.The method that is used for nitrogen atmosphere is imported to the container that comprises product is well known in the art.In other embodiment preferred, also can be when the cooling mixture, the nitrogen air-blowing to mixture, providing oxidation resistant Additional Protection, thereby is improved the oxidation and/or the chemical stability of this product.

In another preferred embodiment, product of the present invention can comprise pharmaceutically useful excipient and/or interpolation the medical active agent (promptly the treatment or active constituents of medicine, and their combination).For the medical active agent that has low solubility in water, this embodiment is particularly advantageous.These pharmaceutical products have the following advantages: the therapeutic activity composition is provided with useful nutrient (for example LC-PUFA).The example of pharmaceutically acceptable excipient is including, but not limited to the aqueous solution, oil, ester and the glycol of the salt solution of water, phosphate buffered, Lin Ge (Ringer ' s) solution, glucose solution, the solution that comprises serum, Hank ' s solution, other physiological equilibrium.Medical active agent of the present invention includes, but not limited to the medicine of inhibin, antihypertensive, antidiabetic, anti-dull-witted medicine, antidepressants, antiadipositas drug, anorectic and raising memory and/or cognitive function.In another preferred embodiment, product of the present invention can comprise COF, for example functional food composition, food additive or other composition.

Product of the present invention can be used alone as food, nutriment or pharmaceutical products, also product of the present invention can be mixed or adds in food, nutriment or the pharmaceutical products.In first embodiment, product of the present invention is the food that comprises oil product of the present invention and food component.Can be with described product directly as COF, for example at beverage, baste (sauces), based on food (for example milk, sour milk, cheese and ice cream) and the oil in the bakery product and/or shortening and/or daubing product and/or other fatty composition of dairy products; Or alternatively as nutriment, for example as nutritional supplement (with the form of capsule or tablet); Meat or product are for the feed or the feed tonic of any animal of people's consumption; The feed or the feed tonic of any companion animals (companionanimal), described companion animals includes but not limited to dog, cat and horse; Food supplementation comprises baby food (baby food) and infant formula.The meaning of term " animal " is any organism that belongs to the animal kingdom, includes but not limited to obtain from it any animal of poultry meat, marine products, beef, pork or lamb.Marine products come from, but are not limited to fish, shrimp and shellfish.Term " product " also comprises any products that comes from these animals except that comprising the meat that comes from these animals, include but not limited to egg, milk or other products.When feeding nutrient (for example LC-PUFAs) to these animals, described nutrient can enter in meat, milk, egg or the other products of these animals, and the content of the described nutrient of these products is improved.In addition, when feeding nutrient (for example LC-PUFAs) to these animals, described nutrient can improve the holistic health of these animals.

Composition of the present invention can be added to multiple product in a plurality of stages of preparation process, for example in bakery product, vitamin tonic, dietary supplements, the beverage powder (powdered drink) etc.Can use composition of the present invention to prepare multiple finished product or half-finished powdered food product.

The part tabulation that comprises the food of the present composition comprises dough, batter, bakes the food class, for example comprises following: cake, cheesecake (cheesecake), group, cup cake, cookies, strip food (bar), bread, volume, biscuit, muffin (muffin), strudel (pastry), flapjack (scone) and a sop in the pan fourth (crouton); Liquid food, for example beverage, energy drink (energy drink), infant formula, liquid meal (liquid meal), fruit juice, contain multivitamin syrup, meal substitute (mealreplacer), medicine food (medicinal food) and syrup; Semisolid food, for example baby food, sour milk, cheese, cereal preparation, pancake mixes material (pancake mix); Be pressed into the food of piece, comprise energy stick (energy bar); Frozen confectionery; The ice yoghourt; China husband compound; The salad flavoring; And for the egg compound.Also comprise bakery product, for example cookies, crispbread (cracker), sweet food (sweetgood), snacks cake (snack cake), group, mixed sweetmeats cake bar/snacks cake bar (granola/snack bar) and baking fermentation food (toaster pastry); Become the flavor snacks, for example potato block (potato chip), cornflakes (corn chip), corn flour sheet (tortilla chip), extruding snacks (extruded snack), puffed rice, pretzels (pretzel), chips (potato crisp) and nut; Special snacks, for example baste (dip), dried fruit snacks, meat flavour snacks, pork rind, the health food that is pressed into piece and rice/corn-dodger; And confectionery snacks, for example candy.

Another product embodiment of the present invention is medical food.The food that medical food comprises is arranged in the preparation that will consume, perhaps under doctor's supervision in the external preparation that gives, and it is intended to be used for the special diet management of disease or illness, based on the principles of science of recognizing, by medical assessment, for described disease or illness are set up distinctive nutritional need.

In this complete introducing U.S. Provisional Patent Application 60/695,996 and U.S. Provisional Patent Application 60/738,304, as a reference.

Although disclose with regard to concrete method, product and organism, but the invention is intended to comprise all such methods, product and the organism that can obtain and use according to the disclosed technology of the application, all that comprise that those skilled in the art can carry out substitute, improve and optimize.For exemplary purposes, provide following examples and result of the test, these embodiment and result of the test are not intended to limit the scope of the invention.

Embodiment

Embodiment 1: prepare high-quality raw oil

In fermentation tank, make the schizochytrium limacinum microorganism belonging to genus growth of being rich in DHA oil, obtain zymotic fluid.Collect zymotic fluid, it is contacted with Alcalase  2.4, Alcalase  2.4 belongs to cytolytic enzyme for making schizochytrium limacinum.The cell mixture of gained dissolving is an emulsion, and it is contacted with 27% aqueous solution of isopropyl alcohol.Mix described mixture by stirring, then that it is centrifugal, obtain the two-phase product of non-emulsification basically.Heavy phase comprises the component of useless zymotic fluid, gently comprises the oil and some isopropyl alcohols and the water that are rich in DHA mutually.Make gently dryly mutually, obtain high-quality raw oil.

Embodiment 2: algae oil is carried out minimum processing

The present embodiment example, the oil of minimum processing procedure produced according to the present invention.

The oil that has prepared minimum processing procedure on a large scale.Under nitrogen, 200 kilograms of high-quality raw oils (preparing as described in embodiment 1) are heated to 65 ℃ to 70 ℃, described high-quality raw oil prepares by the schizochytrium limacinum microorganism belonging to genus, comprises DHA.Then, 50% citric acid soln of about 0.2% ratio of weight of oil (its weight with) is added in the oil, under nitrogen, mixed 30 to 45 minutes.Next, the filter aid of the ratio of weight of oil (its weight with) adds in the oil with 0.2 to 0.5%, filters, so that remove any impurity that is present in the oil.Then, at 210 ℃,, oil is carried out deodorization with 180kg/ hour delivery rate (feed rate).Then, with the additional oil of tocopherol, ascorbyl palmitate and Rosmarinus officinalis extract to deodorization.In table 1, provide the feature of the oil of each treatment step.The meaning of term " PV " is a peroxide number; The meaning of term " FFA " is a free fatty; The meaning of term " p-AV " is the P-anisidine value.The yield of this processing procedure is greater than 98%.

Table 1

Treatment step	PV (meq/kg)	FFA (％)	p-AV	Phosphorus value (ppm)	DHA (％w/w)
Treatment step	PV (meq/kg)	FFA (％)	p-AV	Phosphorus value (ppm)	DHA (％w/w)	Raw oil	0.15	0.22	3.7	3.32	34.0
The acid-treated oil of citron	0.26	0.21	3.6	Be lower than detectability	Do not analyze	Raw oil	0.15	0.22	3.7	3.32	34.0
The acid-treated oil of citron	0.26	0.21	3.6	Be lower than detectability	Do not analyze	The oil of deodorization when not having antioxidant	0.28	0.13	4.9	Be lower than detectability	Do not analyze
The oil of deodorization when having antioxidant	0.0	0.15	4.0	Be lower than detectability	33.2	The oil of deodorization when not having antioxidant	0.28	0.13	4.9	Be lower than detectability	Do not analyze

Embodiment 3a: physics is refining

Get the high-quality raw oil of about 600kg (as described in embodiment 1, preparing) (FFA＜0.3%, phosphorus value＜10ppm, PV＜2meq/kg), under nitrogen and/or vacuum, be heated to 50-55 ℃.Add 50% citric acid of about 0.2% (w/w), under nitrogen and/or vacuum, oil was kept 15 minutes at 50-55 ℃.Add Trisyl 600 (0.1%-3%w/w is generally 0.25%), under nitrogen and/or vacuum, temperature was kept 15 minutes between 50-55 ℃.Add Tonsil Supreme FF bleaching clay (0.1%-4%w/w is less than 0.5% usually), oil is heated to 90-95 ℃, kept 30 minutes in vacuum (＞24 " Hg).Then, add diatomite (0.1-0.5%w/w is generally 0.2%), make oil filter filter through Sparkler.Then, at 210-225 ℃,, oil is carried out deodorization with the flow velocity of 180-225kg/hr.After deodorization, add antioxidant.It is semi-solid oil that this processing procedure obtains in room temperature.

The oily yield of this processing procedure is～92-97%.Table 2 has shown the qualitative data of these steps when antioxidant exists.

Table 2

Test number	Initial FFA (%)	Final FFA (%)	Initial p V (meq/kg)	Final PV (meq/kg)	Initial phosphorus value (ppm)	Final phosphorus value (ppm)
Test number	Initial FFA (%)	Final FFA (%)	Initial p V (meq/kg)	Final PV (meq/kg)	Initial phosphorus value (ppm)	Final phosphorus value (ppm)	Test #1	＜0.1	0.11	1.15	0	9.2	1.9
Test #2	＜0.1	0.09	0.15	0	5.6	0	Test #1	＜0.1	0.11	1.15	0	9.2	1.9
Test #2	＜0.1	0.09	0.15	0	5.6	0	Test #3	0.28	0.19	0.25	＜0.1	2.6	3.4
Test #4	0.23	0.21	0.26	0	3.3	0	Test #3	0.28	0.19	0.25	＜0.1	2.6	3.4

Before adding antioxidant and afterwards, measure the FFAs of the oil of deodorization.After adding antioxidant, observing FFAs increases (about 2 times) significantly.

Embodiment 3b: physics is made with extra care (oil of clarification)

The present embodiment example, the fluid oil of minimum processing procedure produced according to the present invention and relevant solid-state fat products.

Get the high-quality raw oil of about 1200kg (as described in embodiment 1, preparing) (FFA＜0.3%, phosphorus value＜12ppm, PV＜2meq/kg), under nitrogen and/or vacuum, be heated to 50-55 ℃.Add the 50wt% citric acid of about 0.2% (w/w), under nitrogen and/or vacuum, oil was kept 15 minutes at 50-55 ℃.Then, under nitrogen and/or vacuum, utilize different duration (0-12 hour) and agitator speed (4-16rpm), with oil from～55 ℃ be cooled to～35 ℃.Then, add diatomite (0.1-0.5%w/w is generally 0.2%), make oil filter filter through Sparkler.Oil through following processing is repeated described cold filtration step, described being treated to: under nitrogen and/or vacuum, add deep fat, utilize different duration (0-12 hour) and agitator speed (4-16rpm) then, with oil from～50 ℃ be cooled to～30 ℃.Add diatomite (0.1-0.5%w/w is generally 0.2%) once more, make oil filter filter through Sparkler.Next, add Trisyl 600 (0.1%-3%w/w is generally 0.25%), under nitrogen and/or vacuum, temperature was kept 15 minutes between 50-55 ℃.Add TonsilSupreme FF bleaching clay (0.1%-4%w/w, be generally 0.5% or still less), oil is heated to 90-95 ℃, kept 30 minutes in vacuum (＞24 " Hg).Add diatomite (0.1-0.5%w/w is generally 0.2%), make oil filter filter through Sparkler.Then, under nitrogen and/or vacuum, utilize different duration (0-12 hour) and agitator speed (4-16rpm) once more, with oil from～40 ℃ be cooled to～20 ℃.Add diatomite (0.1-0.5%w/w is generally 0.2%), make oil filter filter through Sparkler.Then, at 210-225 ℃,, oil is carried out deodorization with the flow velocity of 180-225kg/hr.After deodorization, add antioxidant.This processing procedure obtains the oil in the room temperature clarification.The oily yield of this processing procedure is～55-60%.Table 3 has shown the qualitative data of these steps when antioxidant exists.

Table 3

Test number	Initial FFA (%)	Final FFA (%)	Initial p V (meq/kg)	Final PV (meq/kg)	Initial phosphorus value (ppm)	Final phosphorus value (ppm)
Test number	Initial FFA (%)	Final FFA (%)	Initial p V (meq/kg)	Final PV (meq/kg)	Initial phosphorus value (ppm)	Final phosphorus value (ppm)	Test #1	0.21	0.1	0.32	0.5	＜5	2.6
Test #2	0.19	0.17	＜0.1	0.07	11	3.1	Test #1	0.21	0.1	0.32	0.5	＜5	2.6
Test #2	0.19	0.17	＜0.1	0.07	11	3.1	Test #3	0.12	0.17	0.53	0.07	3	6.5
Test #4	0.18	0.08	0.26	0	3.3	0.5	Test #3	0.12	0.17	0.53	0.07	3	6.5

Can handle the material that filter is held back, for example by the heating with filtration treatment so that solid matter is separated with bleaching clay.The material that hot filtration apparatus is held back melts solid.Then, can the solid that melt be separated with described clay, solidify again by cooling then for example by filtering.The solid that reclaims can comprise the PUFA of about 20-30%, and wherein major part is DHA.For example the oil of described clarification and solid can be used as food or food additive.

Embodiment 3c: physics is refining/and silica gel is refining

Get the high-quality raw oil of about 100g (as described in embodiment 1, preparing) (FFA＜0.8%, phosphorus value＜10ppm, PV＜2meq/kg), under nitrogen, be heated to 50-55 ℃.Add the 50wt% citric acid of about 0.2% (w/w), under nitrogen and/or vacuum, oil was kept 15 minutes at 50-55 ℃.Next, add the silica gel (Brightsorb F100) of 0.5%-1.25%w/w, under vacuum, oil is heated to 85 ℃.Kept 30 minutes, and added Tonsil Supreme FF bleaching clay (0.5%w/w) afterwards, oil is heated to 90-95 ℃, kept 30 minutes in vacuum (＞24 " Hg).Then, add diatomite (0.1-0.5%w/w is generally 0.2%), be cooled to after 60-65 ℃, use Buchner funnel, oil is carried out vacuum filtration.The yield of these tests is 95-96%.In table 4, show the quality results of these tests.Final products are semisolid oil.Also can carry out deodorization and/or bleaching to this product, it still can be semisolid oil.

Table 4

Test number	% silica gel	Initial FFA (%)	Final FFA (%)	Initial p V (meq/kg)	Final PV (meq/kg)	Initial AV	Final AV
Test number	% silica gel	Initial FFA (%)	Final FFA (%)	Initial p V (meq/kg)	Final PV (meq/kg)	Initial AV	Final AV	Test #1	0.5％	0.64	0.43	1.51	1.40	6.1	n/a
Test #2	0.8％	0.64	0.34	1.51	1.33	6.1	n/a	Test #1	0.5％	0.64	0.43	1.51	1.40	6.1	n/a
Test #2	0.8％	0.64	0.34	1.51	1.33	6.1	n/a	Test #3	1.2％	0.64	0.17	1.51	1.33	6.1	6.3

Embodiment 3d: the alkali refining of improvement

Get about 600kg and contain the high-quality raw oil of 0.8%FFA (as described in embodiment 1, preparing) (phosphorus value＜12ppm, PV＜2meq/kg), under nitrogen and/or vacuum, be heated to 50-55 ℃ at the most.Add the 50wt% citric acid of about 0.2% (w/w), under nitrogen and/or vacuum, oil was kept 15 minutes at 50-55 ℃.After this, 50% caustic alkali (caustic) of 0.1%-0.5%w/w is added in the oil, keep 15-30 minute (the amount ratio standard consumption of caustic alkali few～2-10 doubly) at 60-65 ℃.Then, oil is centrifugal, remove soap-like matter from oil.Add Trisyl 600 (0.1%-3%w/w is generally 0.25%), under nitrogen and/or vacuum, temperature was kept 15 minutes at 50-55 ℃.Add TonsilSupreme FF bleaching clay (0.1%-4%w/w, be generally 0.5% or still less), oil is heated to 90-95 ℃, kept 30 minutes in vacuum (＞24 " Hg).Add diatomite (0.1-0.5%w/w is generally 0.2%), make oil filter filter through Sparkler.Then, at 210-225 ℃,, oil is carried out deodorization with the flow velocity of 180-225kg/hr.After deodorization, add antioxidant.This processing procedure obtains semisolid oil.

The oily yield of this processing procedure is～81-91%.Table 5 has shown the qualitative data of these steps when antioxidant exists.

Table 5

Test number	Initial FFA (%)	Final FFA (%)	Initial p V (meq/kg)	Final PV (meq/kg)	Initial phosphorus value (ppm)	Final phosphorus value (ppm)
Test number	Initial FFA (%)	Final FFA (%)	Initial p V (meq/kg)	Final PV (meq/kg)	Initial phosphorus value (ppm)	Final phosphorus value (ppm)	Test #1	0.26	＜0.1	1.37	0	11.6	4.0
Test #2	0.54	＜0.1	1.84	0	9.8	4.5	Test #1	0.26	＜0.1	1.37	0	11.6	4.0
Test #2	0.54	＜0.1	1.84	0	9.8	4.5	Test #3	0.75	0.1	0.17	＜0.1	8.0	5.0
Test #4	0.40	0.13	0	＜0.1	7.0	0.6	Test #3	0.75	0.1	0.17	＜0.1	8.0	5.0
Test #4	0.40	0.13	0	＜0.1	7.0	0.6	Test #5	0.23	0.08	0.31	0	3.3	0.9

Embodiment 3e: the alkali refining of improvement/do not carry out any centrifugal

Get the high-quality raw oil of about 100g (as described in embodiment 1, preparing) (FFA＜0.3%, phosphorus value＜10ppm, PV＜2meq/kg), under nitrogen and/or vacuum, be heated to 50-55 ℃.Add the 50wt% citric acid of about 0.2% (w/w), under nitrogen and/or vacuum, oil was kept 15 minutes at 50-55 ℃.After this, 50% caustic alkali of 0.4%w/w is added in the oil, keep 15-30 minute (the amount ratio standard consumption of caustic alkali few～2-10 doubly) at 60-65 ℃.Next, add Trisyl600 (1.5%w/w), under nitrogen and/or vacuum, temperature was kept 15 minutes at 50-55 ℃.(0.2%w/w) adds in the oil with diatomite, uses Buchner funnel, and it is carried out vacuum filtration.TonsilSupreme FF bleaching clay (1.0%w/w) is added in the oil of filtration, be heated to 90-95 ℃, kept 30 minutes in vacuum (＞24 " Hg).Add diatomite (0.2%w/w), use Buchner funnel, oil is carried out vacuum filtration.In table 6, show the quality results of this test.Final products are semisolid oil.Also can carry out deodorization and/or bleaching to this product, it still can be semisolid oil.

Table 6

Test number	Initial FFA (%)	Final FFA (%)	Initial p V (meq/kg)	Final PV (meq/kg)	Initial AV	Final AV
Test number	Initial FFA (%)	Final FFA (%)	Initial p V (meq/kg)	Final PV (meq/kg)	Initial AV	Final AV	Test #1	0.64	0.14	1.51	1.21	6.1	5.6

Embodiment 4: the dry fractionation of rough algae oil

The present embodiment example according to the present invention, will become olein fraction and stearin fraction by rough algae oil (algal oil) dry fractionation that comprises DHA that the schizochytrium limacinum microorganism belonging to genus produces.

The 350kg raw oil is carried out dry fractionation of the present invention, so that prepare liquid olein fraction and solid-state stearin fraction.By in container, under stirring is followed, rough algae oil being heated to 60-70 ℃, guarantee that all crystalline phases are all melted in the rough algae oil.Then, during the pre-cooled stage, make material be quickly cooled to 20-30 ℃, agitator speed increases to 40 rpms simultaneously.In order in this stage, to obtain the heat transfer coefficient of maximum possible, use liquid coolant, in the present embodiment, liquid coolant is a water.The temperature of cooling agent does not allow significantly to drop under the nucleation temperature (nucleation temperature).

In stirred vessel, carry out nucleation stage subsequently, by agitator speed being reduced to 20 rpms, make nucleation stage begin to carry out.Be present in the temperature difference between cooling agent and the oil by adjusting, make oil further be reduced to about 12-14 ℃ crystallization temperature from 20-30 ℃ initial oil temperature cooling.In case reached crystallization temperature, just agitator speed be reduced to 15 rpms.For the remaining oil that still is present between the gained crystal, promptly so-called olein fraction after reaching the cloud point of expectation, is transferred to filter element with the gained suspension immediately, stops crystallization process thus.In order to monitor the cloud point of olein fraction, during crystallization stage, the suspension sample is carried out test-filtration.

After crystal suspension being transferred to filter element, liquid phase is extruded, make it through filter cloth.The compression pressure that slowly increases is imposed on the filter chamber, and described compression pressure produces by mechanically reducing filter chamber's volume, and increases lentamente.Final filter pressure reaches 10bar.After filtering, the fraction of separating is weighed.Olein output is the weight of filtrate.Stearin output is the weight that is retained in the crystalline solid on the filter.In table 7, provide the olein fraction of measurement and the yield of stearin fraction.Table 8 has provided the parameter of raw material, olein fraction and stearin fraction.

Table 7

Parameter	The result
Parameter	The result	Cooling curve (h)	13
The final temperature of slurries (℃)	142	Cooling curve (h)	13
The final temperature of slurries (℃)	142	The solid-state fat content (%) of slurries	7.3
The solid-state fat content (%) of stearin	39.6	The solid-state fat content (%) of slurries	7.3
The solid-state fat content (%) of stearin	39.6	Olein yield (%)	83.4
Stearin yield (%)	14.4	Olein yield (%)	83.4

Table 8

Parameter	Raw material	Olein	Stearin
Parameter	Raw material	Olein	Stearin	Water content (ppm)	564-660	-	-
Cloud point (℃)	11.5-17.4	-4.8 to-5.5	-	Water content (ppm)	564-660	-	-
Cloud point (℃)	11.5-17.4	-4.8 to-5.5	-	Iodine number	235.8-265	2604-278.7	184.2-210.8
Fatty acid component (%w/w):				Iodine number	235.8-265	2604-278.7	184.2-210.8
Fatty acid component (%w/w):				12:0	0.2-0.4	0.3-0.4	0.3-0.6
14:0	10.0-12.6	8.6-8.8	14.9-16.1	12:0	0.2-0.4	0.3-0.4	0.3-0.6

Parameter	Raw material	Olein	Stearin
Parameter	Raw material	Olein	Stearin	14:1	0.4-0.5	0.0-0.4	0.5-0.6
16:0	25.3-27.1	22.5-23.1	36.1-39.1	14:1	0.4-0.5	0.0-0.4	0.5-0.6
16:0	25.3-27.1	22.5-23.1	36.1-39.1	16:1	0.7-0.8	0.0	0.0
18:1n-9	0.3-1.9	0.3-0.5	0.0-0.4	16:1	0.7-0.8	0.0	0.0
18:1n-9	0.3-1.9	0.3-0.5	0.0-0.4	22:1	0.9-1.0	1.0-1.1	0.7-0.8
20:5n-3	1.4-1.6	1.7-1.8	1.0-1.5	22:1	0.9-1.0	1.0-1.1	0.7-0.8
20:5n-3	1.4-1.6	1.7-1.8	1.0-1.5	22:5n-6	14.6-17.1	18.0-18.3	11.9-12.9
22:6n-3	39.8-43.4	45.8-46.0	29.1-31.8	22:5n-6	14.6-17.1	18.0-18.3	11.9-12.9
22:6n-3	39.8-43.4	45.8-46.0	29.1-31.8	Solid-state fat content (%):
0℃	8.7	0.0	36.3-44.1	Solid-state fat content (%):
0℃	8.7	0.0	36.3-44.1	10℃	7.5	-	34.8-41.2
15℃	6.8	-	33.2-38.5	10℃	7.5	-	34.8-41.2
15℃	6.8	-	33.2-38.5	20℃	6.1	-	30.5-35.9
25℃	5.4	-	28.9-34.0	20℃	6.1	-	30.5-35.9
25℃	5.4	-	28.9-34.0	30℃	3.1	-	26.3-31.1
35℃	2.4	-	21.0-25.4	30℃	3.1	-	26.3-31.1
35℃	2.4	-	21.0-25.4	40℃	0.8	-	12.9-17.2
45℃	0.0	-	4.5-5.2	40℃	0.8	-	12.9-17.2
45℃	0.0	-	4.5-5.2	50℃	0.0	-	1.5-2.0
55℃	0.0	-	0.0	50℃	0.0	-	1.5-2.0

Can by the application in above embodiment, describe and the minimum processing method of example in any one, or any means by being known in the art, further handle olein (liquid state) fraction and stearin (solid-state or semisolid) fraction, with the oil of preparation deodorization.

Embodiment 5

Following examples show the laboratory scale processing procedure of the solid-state fat products of preparation the present invention.

The oil of the not winterization that will extract from schizochytrium limacinum microorganism belonging to genus living beings with hexane is heated to 40 ℃, all melts until all solid matters, forms the liquid of homogeneous.In another container,, monoglyceride and diglyceride emulsifying agent (DIMODAN PTK A is available from DANISCO) are melted, until becoming liquid fully at identical temperature (40 ℃).Then, the emulsifying agent that melts is added in the oil of thawing, it is mixed.Then, oil/emulsifier mixture is transferred to ice bath, constantly mixes simultaneously in cooling.The product of cooling is solid-state, has the denseness of shortening.Then, with transferred product to container and storage.

Embodiment 6

Following examples are presented at the laboratory scale processing procedure of introducing hydrogen in the curing schedule of product and preparing solid-state fat products.

Described process and above in embodiment 5, discuss identical, different is, when composition cools, with nitrogen with about 10 to about 50ml/min speed air-blowing to composition, this provides extra stability for product, and changed the color/physical property of product, made it to become the yellow appearance of gloss from the outward appearance of dimness.The product of cooling is solid-state, has the denseness of shortening.After cooling, transferred product is to container and storage.

Embodiment 7

Following examples show the pilot-scale or the extensive processing procedure of the solid-state fat products of preparation.

The oil of the not winterization that will extract from schizochytrium limacinum microorganism belonging to genus living beings with hexane is heated to 40 ℃, all melts until all solid matters, forms the liquid of homogeneous.In another container,, the emulsifying agent of embodiment 5 is melted, until becoming liquid fully at identical temperature (40 ℃).Then, the emulsifying agent that melts is added in the oil (HM) of thawing, it is mixed.The liquid emulsion of heat is imported to beaker in ice, stir simultaneously, simulation scraped surface heat exchanger (scrape surface heat exchanger) cools off.Make composition be cooled to about 0 ℃ from about 40 ℃, last about four minutes.During this process, nitrogen is imported in the composition with about speed of 10 to about 50ml/min.After cooling, the gained crystallised fat is transferred to container and storage.

Embodiment 8

Following examples show the pilot-scale or the extensive processing procedure of the solid-state fat products of preparation, the nutrient that described product contains oil and adds.

Described at embodiment 7, the mixture of preparation emulsifying agent and oil.When mixture cools off, constantly mixture is mixed, add water by 5% weight, obtain the slightly different product of denseness.Then, by 5% weight, be that the ascorbic acid of free acid adds in the mixture with form.Then, by 0.0008% weight, folic acid is added in the mixture.Then, the gained crystallised fat is transferred to container and storage.

Embodiment 9

Following examples show, comprise ascorbic acid and comprise ascorbic acid and the oxidation stability of the present composition of folic acid increases.

According to embodiment 5, prepared solid-state fat composition of the present invention.According to embodiment 1, by during cooling step, introducing other composition, prepared the solid-state fat composition that additionally comprises ascorbic acid (5wt.%) and water (5wt.%) and comprise ascorbic acid (5wt.%), folic acid (0.0008wt.%) and water (5wt.%).Estimate the oxidation stability of resulting composition, the result is presented among Fig. 3.As shown in the figure, the interpolation of ascorbic acid and water makes the OSI value of base composition (base composition) increase more than three times.In addition, folic acid is added in the composition that contains ascorbic acid and water, the oxidation stability of composition is improved.

Embodiment 10: the blend composition that comprises the oil of minimum processing procedure

Use ARASCO ^(Martek Biosciences, Columbia MD), with 2: 1 ratio, bleach the oil that is rich in DHA that comes from above embodiment 3a and 3d, and preparation comprises the oil mixture of ARA and DHA.

DHA and ARASCO from minimum processing procedure ^The qualitative character of the blend compositions of preparation

Parameter	ARASCO ^/ DHA oil (embodiment 3a), 2: 1	ARASCO ^/ DHA oil (embodiment 3d), 2: 1
Parameter	ARASCO ^/ DHA oil (embodiment 3a), 2: 1	ARASCO ^/ DHA oil (embodiment 3d), 2: 1	Physics is described:
Color (perusal)	Faint yellow	Orange	Physics is described:
Color (perusal)	Faint yellow	Orange	Color (Lovibond colour (Lovibond))	1.9R/70.0Y	3.2R/70.0Y
Breakdown of oil degree at 25 ℃	The opaque liquid of thickness	The opaque liquid of thickness	Color (Lovibond colour (Lovibond))	1.9R/70.0Y	3.2R/70.0Y
Breakdown of oil degree at 25 ℃	The opaque liquid of thickness	The opaque liquid of thickness	Breakdown of oil degree at 40 ℃	The liquid of clarification	The liquid of clarification
Chemical analysis:			Breakdown of oil degree at 40 ℃	The liquid of clarification	The liquid of clarification
Chemical analysis:			DHA(mg/g)	123	127
DHA (area percentage, %)	12.9	13.3	DHA(mg/g)	123	127
DHA (area percentage, %)	12.9	13.3	ARA(mg/g)	247.3	255.6
ARA (area percentage, %)	27.3	28.1	ARA(mg/g)	247.3	255.6
ARA (area percentage, %)	27.3	28.1	PV(meq/kg)	0.45	0.45
p-AV	5.9	7.2	PV(meq/kg)	0.45	0.45
p-AV	5.9	7.2	FFA(％)	0.07	0.07
Moisture and volatile matter (%)	Be lower than detectability	Be lower than detectability	FFA(％)	0.07	0.07
Moisture and volatile matter (%)	Be lower than detectability	Be lower than detectability	Saponifiable matter (%) not	1.9	1.9
Trans-fatty acid (%)	Be lower than detectability	Be lower than detectability	Saponifiable matter (%) not	1.9	1.9
Trans-fatty acid (%)	Be lower than detectability	Be lower than detectability	Elementary analysis:
As	Be lower than detectability	Be lower than detectability	Elementary analysis:
As	Be lower than detectability	Be lower than detectability	Cu	Be lower than detectability	Be lower than detectability
Fe	Be lower than detectability	Be lower than detectability	Cu	Be lower than detectability	Be lower than detectability
Fe	Be lower than detectability	Be lower than detectability	Pb	Be lower than detectability	Be lower than detectability
Hg	Be lower than detectability	Be lower than detectability	Pb	Be lower than detectability	Be lower than detectability
Hg	Be lower than detectability	Be lower than detectability	Fatty acid profile (main aliphatic acid):
14:0	5.2	4.0	Fatty acid profile (main aliphatic acid):
14:0	5.2	4.0	16:0	17.4	16.7
18:0	5.7	5.8	16:0	17.4	16.7

18:1n-9	10.9	10.9
18:1n-9	10.9	10.9	18:2n-6	4.6	4.8
18:3n-6	1.9	2.0	18:2n-6	4.6	4.8
18:3n-6	1.9	2.0	22:0	1.0	1.0
20:3n-6	2.3	2.3	22:0	1.0	1.0
20:3n-6	2.3	2.3	20:4n-6	27.3	28.1
24:0	1.1	1.1	20:4n-6	27.3	28.1
24:0	1.1	1.1	22:5n-6	4.8	5.0
22:6n-3	12.9	13.3	22:5n-6	4.8	5.0

Embodiment 11

Following examples show the laboratory scale processing procedure of the solid-state fat products of preparation, and described product contains oil and the palm stearines that comes from the schizochytrium limacinum genus.

Under nitrogen, will be heated to 40-50 ℃ from the refining fully oil of the not winterization of schizochytrium limacinum microorganism belonging to genus living beings preparations, all melt until all solid matters, form the liquid of homogeneous.In another container, at identical temperature (40-50 ℃), (available from Ciranda Inc., Hudson WI) melts, until becoming liquid fully to make palm stearines.The oil of used not winterization and the ratio of palm stearines be 75 to 25 (%, w/w).Next, the palm stearines of thawing is mixed with the oil of the not winterization that comes from the schizochytrium limacinum genus.In another container, (Dimodan 930-KA or Grindsted PS 219/B K-A available from Danisco, Denmark) are heated to 70-75 ℃, until the liquid that forms homogeneous with monoglyceride and diglyceride emulsifying agent.Then, the oil mixture (the not oil of winterization and palm stearines) that melts is added in the emulsifying agent of thawing, it is mixed.In cooler (chiller batch) in batches,, make the liquid formulation cooling of heat be reduced to 15 ℃ at nitrogen with under stirring.In case reach crystallization temperature, formulation was kept 1 hour at 15 ℃, stir simultaneously.Then, gained crystallised fat formulation is transferred to container and storage.The results are shown in the table 9.

Table 9

Physics and chemical property	The result
Physics and chemical property	The result	Peroxide number (meq/kg)	4.1-8.6
Free fatty (%)	0.15-0.17	Peroxide number (meq/kg)	4.1-8.6
Free fatty (%)	0.15-0.17	The p-anisidine value	Be lower than detectability
Rancimat(hr)	10.9-13.5	The p-anisidine value	Be lower than detectability
Rancimat(hr)	10.9-13.5	DHA content (mg/g)	215.4-239.2
Solid-state fat content (%):		DHA content (mg/g)	215.4-239.2
Solid-state fat content (%):		10.0℃	24.4-26.9
21.1℃	18.3-19.8	10.0℃	24.4-26.9
21.1℃	18.3-19.8	26.7℃	14.6-16.6
33.3℃	9.2-10.6	26.7℃	14.6-16.6

37.8℃

7.8-8.1

Embodiment 12

Following examples show the laboratory scale processing procedure of the solid-state fat products of preparation, and described product contains oil and the palm kernel stearin that comes from the schizochytrium limacinum genus.

Under nitrogen, will be heated to 40-50 ℃ from the refining fully oil of the not winterization of schizochytrium limacinum microorganism belonging to genus living beings preparations, all melt until all solid matters, form the liquid of homogeneous.In another container, at identical temperature (40-50 ℃), (available from Ciranda Inc., Hudson WI) melts, until becoming liquid fully to make the palm kernel stearin.Not the ratio of the oil of winterization and palm kernel stearin be 75: 25 to 80: 20 (%, w/w).Next, the palm kernel stearin of thawing is mixed with the oil of the not winterization that comes from the schizochytrium limacinum genus.In another container, (Dimodan930-KA or Grindsted PS 219/B K-A available from Danisco, Denmark) are heated to 70-75 ℃, until the liquid that forms homogeneous with monoglyceride and diglyceride emulsifying agent.Then, the oil mixture (the not oil of winterization and palm kernel stearin) that melts is added in the emulsifying agent of thawing, it is mixed.In cooler in batches,, make the liquid formulation cooling of heat be reduced to 15 ℃ at nitrogen with under stirring.In case reached crystallization temperature, formulation was kept 1 hour at 15 ℃, stir simultaneously.Then, gained crystallised fat formulation is transferred to container and storage.The results are shown in the table 10.

Table 10

Physics and chemical property	The result
Physics and chemical property	The result	Peroxide number (meq/kg)	1.1
Free fatty (%)	0.11	Peroxide number (meq/kg)	1.1
Free fatty (%)	0.11	The p-anisidine value	Be lower than detectability
Rancimat(hr)	7.3	The p-anisidine value	Be lower than detectability
Rancimat(hr)	7.3	DHA content (mg/g)	225.8
Solid-state fat content (%):		DHA content (mg/g)	225.8
Solid-state fat content (%):		10.0℃	32.6
21.1℃	18.3	10.0℃	32.6
21.1℃	18.3	26.7℃	11.2
33.3℃	4.1	26.7℃	11.2
33.3℃	4.1	37.8℃	2.0

Embodiment 13

Following examples show the laboratory scale processing procedure of the solid-state fat products of preparation, and described product comprises the antioxidant that schizochytrium limacinum belongs to oil and palm kernel stearin and interpolation.

To mix from refining fully oil and 0.2% (w/w) antioxidant (comprising 10% tocopherol and 10% ascorbyl palmitate) of the not winterization of schizochytrium limacinum microorganism belonging to genus living beings preparations, under nitrogen, the gained mixture is heated to 40-50 ℃, all melt until all solid matters, form the liquid of homogeneous.In another container, at identical temperature (40-50 ℃), (available from Ciranda Inc., Hudson WI) melts, until becoming liquid fully to make the palm kernel stearin.The ratio of the oil of winterization and palm kernel stearin was not by (%, w/w) use in 75: 25.Next, the palm kernel stearin of thawing is mixed with the oil of the not winterization that comes from the schizochytrium limacinum genus.In another container, (Dimodan 930-KA or Grindsted PS 219/B K-A available from Danisco, Denmark) are heated to 70-75 ℃, until the liquid that forms homogeneous with monoglyceride and diglyceride emulsifying agent.Then, the oil mixture (the not oil of winterization and palm kernel stearin) that melts is added in the emulsifying agent of thawing, it is mixed.In cooler in batches,, make the liquid formulation cooling of heat be reduced to 15 ℃ at nitrogen with under stirring.In case reached crystallization temperature, just made formulation follow maintenance down 1 hour with stirring at 15 ℃.Then, gained crystallised fat formulation is transferred to container and storage.The results are shown in the table 11.

Table 11

Physics and chemical property	The result
Physics and chemical property	The result	Peroxide number (meq/kg)	0.3-0.5
Free fatty (%)	0.15-0.20	Peroxide number (meq/kg)	0.3-0.5
Free fatty (%)	0.15-0.20	The p-anisidine value	Be lower than detectability
Rancimat(hr)	18.9-25.0	The p-anisidine value	Be lower than detectability
Rancimat(hr)	18.9-25.0	DHA content (mg/g)	232-243
Solid-state fat content (%):		DHA content (mg/g)	232-243
Solid-state fat content (%):		10.0℃	30.6-35.6
21.1℃	16.6-21.9	10.0℃	30.6-35.6
21.1℃	16.6-21.9	26.7℃	9.7-15.2
33.3℃	2.3-4.2	26.7℃	9.7-15.2
33.3℃	2.3-4.2	37.8℃	1.5-3.2

Embodiment 14

Following examples show the laboratory scale processing procedure of the solid-state fat products of preparation, and described product contains schizochytrium limacinum and belongs to oil and palm kernel stearin, comprises 10-20% (w/w) DHA.

To mix from refining fully oil and 0.2% (w/w) antioxidant (comprising 10% tocopherol and 10% ascorbyl palmitate) of the not winterization of schizochytrium limacinum microorganism belonging to genus living beings preparations, under nitrogen, the gained mixture is heated to 40-50 ℃, all melt until all solid matters, form the liquid of homogeneous.In another container, at identical temperature (40-50 ℃), (available from Ciranda Inc., Hudson WI) melts, until becoming liquid fully to make the palm kernel stearin.The ratio of the oil of winterization and palm kernel stearin was not by (%, w/w) use in 60: 40.Next, the palm kernel stearin of thawing is mixed with the oil of the not winterization that comes from the schizochytrium limacinum genus.In another container, (Dimodan 930-KA or Grindsted PS 219/B K-A available from Danisco, Denmark) are heated to 70-75 ℃, until the liquid that forms homogeneous with monoglyceride and diglyceride emulsifying agent.Then, the oil mixture (the not oil of winterization and palm kernel stearin) that melts is added in the emulsifying agent of thawing, it is mixed.In a cooler batch,, make the liquid formulation cooling of heat be reduced to 15 ℃ at nitrogen with under stirring.In case reached crystallization temperature, just made formulation follow maintenance down 1 hour with stirring at 15 ℃.Then, gained crystallised fat formulation is transferred to container and storage.The results are shown in the table 12.

Table 12

Physics and chemical property	The result
Physics and chemical property	The result	Peroxide number (meq/kg)	0.5-0.7
Free fatty (%)	0.23-0.25	Peroxide number (meq/kg)	0.5-0.7
Free fatty (%)	0.23-0.25	The p-anisidine value	Be lower than detectability
Rancimat(hr)	20-26	The p-anisidine value	Be lower than detectability
Rancimat(hr)	20-26	DHA content (mg/g)	192-199
Solid-state fat content (%):		DHA content (mg/g)	192-199
Solid-state fat content (%):		10.0℃	39.8-42.6
21.1℃	25.0-27.1	10.0℃	39.8-42.6
21.1℃	25.0-27.1	26.7℃	13.6-12.2
33.3℃	2.8-2.9	26.7℃	13.6-12.2
33.3℃	2.8-2.9	37.8℃	2.5-3.0

Embodiment 15

Following examples show the laboratory scale processing procedure of the solid-state fat products of preparation, and described product contains schizochytrium limacinum and belongs to oil and palm kernel stearin, comprises 20-30% (w/w) DHA.

To mix from refining fully oil and 0.2% (w/w) antioxidant (comprising 10% tocopherol and 10% ascorbyl palmitate) of the not winterization of schizochytrium limacinum microorganism belonging to genus living beings preparations, under nitrogen, the gained mixture is heated to 40-50 ℃, all melt until all solid matters, form the liquid of homogeneous.In another container, at identical temperature (40-50 ℃), (available from Ciranda Inc., Hudson WI) melts, until becoming liquid fully to make the palm kernel stearin.Not the ratio of the oil of winterization and palm kernel stearin be 75: 25 to 85: 15 (%, w/w).Next, the palm kernel stearin of thawing is mixed with the oil of the not winterization that comes from the schizochytrium limacinum genus.In another container, (Dimodan 930-KA or Grindsted PS 219/B K-A available from Danisco, Denmark) are heated to 70-75 ℃, until the liquid that forms homogeneous with monoglyceride and diglyceride emulsifying agent.Then, the oil mixture (the not oil of winterization and palm kernel stearin) that melts is added in the emulsifying agent of thawing, it is mixed.In a cooler batch,, make the liquid formulation cooling of heat be reduced to 15 ℃ at nitrogen with under stirring.In case reached crystallization temperature, just made formulation follow maintenance down 1 hour with stirring at 15 ℃.Then, gained crystallised fat formulation is transferred to container and storage.The results are shown in the table 13.

Table 13

Physics and chemical property	The result
Physics and chemical property	The result	Peroxide number (meq/kg)	0.0-0.6
Free fatty (%)	0.16-0.24	Peroxide number (meq/kg)	0.0-0.6
Free fatty (%)	0.16-0.24	The p-anisidine value	0.0-5.0
Rancimat(hr)	19.6-22.4	The p-anisidine value	0.0-5.0
Rancimat(hr)	19.6-22.4	DHA content (mg/g)	236-283
Solid-state fat content (%):		DHA content (mg/g)	236-283
Solid-state fat content (%):		10.0℃	27.7-32.1
21.1℃	15.2-18.1	10.0℃	27.7-32.1
21.1℃	15.2-18.1	26.7℃	9.8-11.6
33.3℃	4.5-4.9	26.7℃	9.8-11.6
33.3℃	4.5-4.9	37.8℃	2.7-2.9

Embodiment 16

Following examples show the laboratory scale processing procedure of the solid-state fat products of preparation, and described product comprises schizochytrium limacinum and belongs to oil and palm kernel stearin, contains＞30% (w/w) DHA.

To mix from refining fully oil and 0.2% (w/w) antioxidant (10% tocopherol and 10% ascorbyl palmitate) of the not winterization of schizochytrium limacinum microorganism belonging to genus living beings preparations, under nitrogen, the gained mixture is heated to 40-50 ℃, all melt until all solid matters, form the liquid of homogeneous.In another container, at identical temperature (40-50 ℃), (available from CirandaInc., Hudson WI) melts, until becoming liquid fully to make the palm kernel stearin.The oil of used not winterization and the ratio of palm kernel stearin be 80: 20 (%, w/w).Next, the palm kernel stearin of thawing is mixed with the oil of the not winterization that comes from the schizochytrium limacinum genus.In another container, (Dimodan 930-KA or Grindsted PS 219/B K-A available from Danisco, Denmark) are heated to 70-75 ℃, until the liquid that forms homogeneous with monoglyceride and diglyceride emulsifying agent.Then, the oil mixture (the not oil of winterization and palm kernel stearin) that melts is added in the emulsifying agent of thawing, it is mixed.In a cooler batch,, make the liquid formulation cooling of heat be reduced to 15 ℃ at nitrogen with under stirring.In case reached crystallization temperature, just made formulation follow maintenance down 1 hour with stirring at 15 ℃.Then, gained crystallised fat formulation is transferred to container and storage.The results are shown in the table 14.

Table 14

Physics and chemical property	The result
Physics and chemical property	The result	Peroxide number (meq/kg)	0.2-0.4
Free fatty (%)	0.12-0.17	Peroxide number (meq/kg)	0.2-0.4
Free fatty (%)	0.12-0.17	The p-anisidine value	Be lower than detectability
Rancimat(hr)	18.9-19.9	The p-anisidine value	Be lower than detectability
Rancimat(hr)	18.9-19.9	DHA content (mg/g)	310-319
Solid-state fat content (%):		DHA content (mg/g)	310-319
Solid-state fat content (%):		10.0℃	23.1-28.5
21.1℃	11.4-16.5	10.0℃	23.1-28.5
21.1℃	11.4-16.5	26.7℃	6.3-10.7
33.3℃	4.3-5.0	26.7℃	6.3-10.7
33.3℃	4.3-5.0	37.8℃	2.9-3.1

Embodiment 17

Following examples show the laboratory scale processing procedure of the solid-state fat products of preparation, described product comprises schizochytrium limacinum and belongs to oil and palm kernel stearin, contains the natural butter aroma (NaturalButter Flavor) and the bitter taste smoke agent for shielding (Bitterness Masker) of interpolation.

To mix from refining fully oil and 0.2% (w/w) antioxidant (10% tocopherol and 10% ascorbyl palmitate), the natural butter aroma of 0.1-0.15% (w/w) (available from Danisco) and 0.03-0.05% (w/w) the natural bitterness smoke agent for shielding (available from Firmenich Inc.) of the not winterization of schizochytrium limacinum microorganism belonging to genus living beings preparations.Then, under nitrogen, the gained mixture is heated to 40-50 ℃, all melts, form the liquid of homogeneous until all solid matters.In another container, at identical temperature (40-50 ℃), (available from Ciranda Inc., Hudson WI) melts, until becoming liquid fully to make the palm kernel stearin.The oil of used not winterization and the ratio of palm kernel stearin be 80: 20 (%, w/w).Next, the palm kernel stearin of thawing is mixed with the oil of the not winterization that comes from the schizochytrium limacinum genus.In another container, (Dimodan 930-KA or GrindstedPS 219/B K-A available from Danisco, Denmark) are heated to 70-75 ℃, until the liquid that forms homogeneous with monoglyceride and diglyceride emulsifying agent.Then, the oil mixture (the not oil of winterization and palm kernel stearin) that melts is added in the emulsifying agent of thawing, it is mixed.In a cooler batch,, make the liquid formulation cooling of heat be reduced to 15 ℃ at nitrogen with under stirring.In case reached crystallization temperature, just made formulation follow maintenance down 1 hour with stirring at 15 ℃.Then, gained crystallised fat formulation is transferred to container and storage.The results are shown in the table 15.

Table 15

Physics and chemical property	The result
Physics and chemical property	The result	Peroxide number (meq/kg)	Be lower than detectability
Free fatty (%)	0.19-0.2	Peroxide number (meq/kg)	Be lower than detectability
Free fatty (%)	0.19-0.2	The p-anisidine value	Be lower than detectability
Rancimat(hr)	28.3-28.5	The p-anisidine value	Be lower than detectability
Rancimat(hr)	28.3-28.5	DHA content (mg/g)	230-283
Solid-state fat content (%):		DHA content (mg/g)	230-283
Solid-state fat content (%):		10.0℃	21.8-27.9
21.1℃	14.0-14.8	10.0℃	21.8-27.9
21.1℃	14.0-14.8	26.7℃	7.2-8.6
33.3℃	2.6-4.0	26.7℃	7.2-8.6
33.3℃	2.6-4.0	37.8℃	1.9-2.2

Principle of the present invention, embodiment preferred and mode of operation have been described in the above-mentioned specification.Yet, should not be interpreted as being limited to disclosed concrete form in the invention of this intention protection, because these concrete forms are considered to illustrative, and not restrictive.Those skilled in the art can be in the case without departing from the gist of the present invention, and the present invention is changed and changes.Therefore, the above-mentioned specific embodiment of the present invention should be regarded as having exemplary in nature, and should not be regarded as limiting the scope of the invention and the purport that appended claims is set forth.

Claims

1. prepare the method for solid-state fat composition, comprising:

A) will comprise the oil and at least a emulsifier of saturated fat and microbial oil, and form mixture, described microbial oil comprises at least a LC-PUFA; And

B) make described mixture solidified, form solid-state fat composition.

2. the process of claim 1 wherein that described oil comprises the LC-PUFA and about 20wt.% saturated fat to about 60wt.% of about 5wt.% to about 70wt.%.

3. the method for claim 2, wherein said saturated fat are not that external source is added.

4. the method for claim 2, wherein said saturated fat are that external source is added.

5. the process of claim 1 wherein that described microbial oil is a winterization not.

6. the process of claim 1 wherein that described oil is unhydrided.

7. the method for claim 1, wherein said microbial oil comes from and is selected from following microorganism: the microorganism that the microorganism that the microorganism that the microorganism that the microorganism that the microorganism that the microorganism of genus thraustochytrium, schizochytrium limacinum belong to, Althornia belong to, Aplanochytrium belong to, Japonochytrium belong to, Elina belong to, Crypthecodinium belong to, and their mixture.

8. the method for claim 7, wherein said microorganism are selected from the microorganism of genus thraustochytrium, the microorganism that schizochytrium limacinum belongs to, the microorganism that Crypthecodinium belongs to, and their mixture.

9. the process of claim 1 wherein that described microbial oil comprises that carbon chain lengths is at least 20 LC-PUFA.

10. the method for claim 9, the carbon chain lengths of wherein said LC-PUFA is at least 22.

11. the method for claim 9, wherein said LC-PUFA has at least three two keys.

12. the method for claim 9, wherein said LC-PUFA has at least four two keys.

13. the method for claim 9, wherein said LC-PUFA comprises DHA.

14. the method for claim 13, wherein said oil comprises the DHA at least about 50% weight.

15. the method for claim 13, wherein said oil comprises the DHA at least about 60% weight.

16. the method for claim 9, wherein said LC-PUFA comprises clupanodonic acid.

17. the method for claim 9, wherein said LC-PUFA comprises arachidonic acid.

18. the method for claim 9, wherein said LC-PUFA comprises eicosapentaenoic acid.

19. the process of claim 1 wherein that described solid-state fat composition has the structure of homogeneous.

20. the process of claim 1 wherein that described solid-state fat composition is a shortening.

21. the method for claim 1; wherein said emulsifying agent is selected from monoglyceride, diglyceride, monoglyceride/diglyceride combination, lecithin, dilactic acid monoglyceride-diglyceride, polyglycerol ester, sucrose fatty ester, stearyl dilactic acid sodium, stearyl dilactic acid calcium, and their combination.

22. the method for claim 21, wherein said emulsifying agent are monoglyceride/diglyceride combination.

23. the process of claim 1 wherein that the amount of described emulsifying agent is that about 0.01% weight is to about 2.0% weight.

24. the process of claim 1 wherein that the amount of described emulsifying agent is that about 0.05% weight is to about 0.2% weight.

25. the process of claim 1 wherein that described solid-state fat composition has the melt temperature at least about 20 ℃.

26. the process of claim 1 wherein that described solid-state fat composition has the melt temperature at least about 30 ℃.

27. the process of claim 1 wherein that described solid-state fat composition has the melt temperature at least about 35 ℃.

28. the process of claim 1 wherein that the described step that makes described mixture solidified controls the formation of crystal in the described solid-state fat composition.

29. the method for claim 28, wherein said crystal comprises β ' crystal.

30. the method for claim 29, wherein described fat in described solid-state fat composition and/or oil is β ' crystalline form at least about 50wt.%.

31. the method for claim 29, wherein described fat in described solid-state fat composition and/or oil is β ' crystalline form at least about 80wt.%.

32. the process of claim 1 wherein and heat described oil.

33. the method for claim 32 wherein before described blend step, heats described oil.

34. the method for claim 32 wherein is heated to described oil at least about 40 ℃.

35. the process of claim 1 wherein and heat described emulsifying agent.

36. the method for claim 35 wherein before described blend step, heats described emulsifying agent.

37. the method for claim 35 wherein is heated to described emulsifying agent at least about 40 ℃.

38. the process of claim 1 wherein that described mixed step comprises the described mixture of stirring.

39. the method for claim 38, wherein said whipping step forms continuous mixture.

40. the process of claim 1 wherein that the described step that makes described mixture solidified comprises makes described mixture cooling.

41. the method for claim 40, wherein said cooling step comprise make described mixture be cooled to about 0 ℃ to about 3 ℃ temperature.

42. the method for claim 40, wherein said curing schedule are mixed described mixture during also being included in described cooling step.

43. the method for claim 40 is wherein cooled off described mixture with about 1 ℃/min to the speed of about 20 ℃/min.

44. the process of claim 1 wherein that described curing schedule comprises imports to nitrogen in the described mixture.

45. the method for claim 44, wherein said importing step comprise the nitrogen air-blowing through described mixture.

46. the method for claim 1 also comprises at least a other composition is added in the described mixture.

47. the method for claim 46, wherein said other composition is a water-soluble liquid.

48. the method for claim 47, wherein said water-soluble liquid are water.

49. the method for claim 47, wherein said water-soluble liquid is by the amount interpolation of about 1wt.% to about 10wt.%.

50. the method for claim 46, wherein said other composition is selected from antioxidant, spices, flavoring agent, sweetener, pigment, vitamin, mineral matter, preceding biologic artifact, probio compound, therapeutic component, drug ingedient, functional food composition, processing composition, and their combination.

51. the method for claim 46, wherein said other composition are the salt of ascorbic acid or ascorbic acid.

52. the method for claim 51, the salt of wherein said ascorbic acid or ascorbic acid is by the amount interpolation of about 0.5wt.% to about 5wt.%.

53. the method for claim 46, wherein said other composition is an antioxidant.

54. the method for claim 53, wherein said antioxidant is selected from ascorbyl palmitate, tocopherol, citric acid, ascorbic acid, tertiary butylated hydroquinone, Rosmarinus officinalis extract, lecithin, and their mixture.

55. the process of claim 1 wherein that described solid-state fat composition has the OSI value at least about 20.

56. the process of claim 1 wherein that described solid-state fat composition has the OSI value at least about 40.

57. the process of claim 1 wherein that described solid-state fat composition has the OSI value at least about 60.

58. the process of claim 1 wherein that described solid-state fat composition is selected from food, nutriment and pharmaceutical products.

59. the method for claim 1 also comprises described solid-state fat composition is added in the product that is selected from food, nutriment and the pharmaceutical products.

60. a solid-state fat composition comprises the not winterization microbial oil that contains LC-PUFA and the mixture of emulsifying agent, wherein said mixture is a solid-state composition in room temperature.

61. the solid-state fat composition of claim 60, wherein said oil comprises saturated fat.

62. not being external sources, the solid-state fat composition of claim 60, wherein said saturated fat do not add.

63. being external sources, the solid-state fat composition of claim 60, wherein said saturated fat add.

64. the solid-state fat composition of claim 60, wherein said oil is unhydrided.

65. the solid-state fat composition of claim 60, wherein said microbial oil comes from and is selected from following microorganism: the microorganism that the microorganism that the microorganism that the microorganism that the microorganism that the microorganism that the microorganism of genus thraustochytrium, schizochytrium limacinum belong to, Althornia belong to, Aplanochytrium belong to, Japonochytrium belong to, Elina belong to, Crypthecodinium belong to, and their mixture.

66. the solid-state fat composition of claim 65, wherein said microorganism are selected from the microorganism of genus thraustochytrium, the microorganism that schizochytrium limacinum belongs to, the microorganism that Crypthecodinium belongs to, and their mixture.

67. the solid-state fat composition of claim 60, wherein said microbial oil comprise that carbon chain lengths is at least 20 LC-PUFA.

68. the solid-state fat composition of claim 60, the carbon chain lengths of wherein said LC-PUFA is at least 22.

69. the solid-state fat composition of claim 60, wherein said LC-PUFA have at least three two keys.

70. the solid-state fat composition of claim 60, wherein said LC-PUFA have at least four two keys.

71. the solid-state fat composition of claim 60, wherein said LC-PUFA comprises DHA.

72. the solid-state fat composition of claim 60, wherein said oil comprises the DHA at least about 50% weight.

73. the solid-state fat composition of claim 60, wherein said oil comprises the DHA at least about 60% weight.

74. the solid-state fat composition of claim 60, wherein said LC-PUFA comprises clupanodonic acid.

75. the solid-state fat composition of claim 60, wherein said LC-PUFA comprises arachidonic acid.

76. the solid-state fat composition of claim 60, wherein said LC-PUFA comprises eicosapentaenoic acid.

77. the solid-state fat composition of claim 60, wherein said solid-state fat composition is a shortening.

78. the solid-state fat composition of claim 60; wherein said emulsifying agent is selected from monoglyceride, diglyceride, monoglyceride/diglyceride combination, lecithin, dilactic acid monoglyceride-diglyceride, polyglycerol ester, sucrose fatty ester, stearyl dilactic acid sodium, stearyl dilactic acid calcium, and their combination.

79. the solid-state fat composition of claim 60, wherein said emulsifying agent are monoglyceride/diglyceride combination.

80. the solid-state fat composition of claim 60, the amount of wherein said emulsifying agent are that about 0.01% weight is to about 2.0% weight.

81. the solid-state fat composition of claim 60, the amount of wherein said emulsifying agent are that about 0.05% weight is to about 0.2% weight.

82. the solid-state fat composition of claim 60, wherein said composition comprises crystal.

83. the solid-state fat composition of claim 82, wherein said crystal comprises β ' crystal.

84. the solid-state fat composition of claim 82, wherein described fat in described solid-state fat composition and/or oil is β ' crystalline form at least about 50wt.%.

85. the solid-state fat composition of claim 82, wherein described fat in described solid-state fat composition and/or oil is β ' crystalline form at least about 80wt.%.

86. the solid-state fat composition of claim 60 also comprises at least a other composition.

87. the solid-state fat composition of claim 86, wherein said other composition is a water-soluble liquid.

88. the solid-state fat composition of claim 87, wherein said water-soluble liquid are water.

89. the solid-state fat composition of claim 87, the amount of wherein said water-soluble liquid are that about 1wt.% is to about 10wt.%.

90. the solid-state fat composition of claim 86, wherein said other composition is selected from antioxidant, spices, flavoring agent, sweetener, pigment, vitamin, mineral matter, preceding biologic artifact, probio compound, therapeutic component, drug ingedient, functional food composition, processing composition, and their combination.

91. the solid-state fat composition of claim 86, wherein said other composition are the salt of ascorbic acid or ascorbic acid.

92. the solid-state fat composition of claim 91, the salt of wherein said ascorbic acid or ascorbic acid is by the amount interpolation of about 0.5wt.% to about 5wt.%.

93. the solid-state fat composition of claim 86, wherein said other composition is an antioxidant.

94. the solid-state fat composition of claim 93, wherein said antioxidant is selected from ascorbyl palmitate, tocopherol, citric acid, ascorbic acid, tertiary butylated hydroquinone, Rosmarinus officinalis extract, lecithin, and their mixture.

95. the solid-state fat composition of claim 60, wherein said composition have the OSI value at least about 20.

96. the solid-state fat composition of claim 60, wherein said composition have the OSI value at least about 40.

97. the solid-state fat composition of claim 60, wherein said composition have the OSI value at least about 60.

98. a fat composition comprises

A) microbial oil of winterization not, described microbial oil comprise that about 5wt.% is to the LC-PUFA of about 70wt.% and the about 20wt.% saturated fat of about 60wt.% extremely; And

B) about 0.01wt.% is to the emulsifying agent of about 2.0wt.%;

Wherein said composition comprises the water that is less than about 10wt.%, and wherein said composition is a solid-state composition in room temperature.

99. the method for the oil product that preparation is used to consume comprises:

A) extract the oil-containing fraction from microbial biomass, wherein said oil-containing fraction comprises at least a LC-PUFA and is enough to the saturated fatty acid of the described oil-containing fraction of visual impact at least;

B) handle described oil-containing fraction by vacuum evaporation, comprise the oil product of at least a LC-PUFA with preparation;

Wherein said oil product does not experience the winterization step.

100. the method for claim 99, wherein said oil product does not experience alkali refining process.

101. the method for claim 99, wherein said oil product does not experience the cold filtration process.

102. the method for claim 99, wherein said oil product does not experience bleaching process.

103. the method for claim 99, wherein said oil product does not experience alkali refining process, cold filtration process or bleaching process.

104. the method for claim 99, wherein said microbial biomass comes from and is selected from following microorganism: the microorganism that the microorganism that the microorganism that the microorganism that the microorganism that the microorganism that the microorganism of genus thraustochytrium, schizochytrium limacinum belong to, Althornia belong to, Aplanochytrium belong to, Japonochytrium belong to, Elina belong to, Crypthecodinium belong to, and their mixture.

105. the method for claim 104, wherein said microorganism are selected from the microorganism of schizochytrium limacinum genus, the microorganism that Crypthecodinium belongs to, and their mixture.

106. the method for claim 99, wherein said oil-containing fraction comprise that carbon chain lengths is at least 20 LC-PUFA.

107. the method for claim 106, the carbon chain lengths of wherein said LC-PUFA is at least 22.

108. the method for claim 106, wherein said LC-PUFA has at least three two keys.

109. the method for claim 106, wherein said LC-PUFA has at least four two keys.

110. the method for claim 106, wherein said LC-PUFA comprises DHA.

111. the method for claim 106, wherein said LC-PUFA comprises clupanodonic acid.

112. the method for claim 106, wherein said LC-PUFA comprises arachidonic acid.

113. the method for claim 106, wherein said LC-PUFA comprises eicosapentaenoic acid.

114. the method for claim 99, the step of the described oil-containing fraction of wherein said processing comprises the precipitation thinner.

115. the method for claim 114, wherein said precipitation thinner are included in the oil-containing fraction that high temperature puts on vacuum condition described extraction.

116. the method for claim 114, wherein said high temperature are about 50 ℃ to about 70 ℃.

117. the method for claim 115, wherein said precipitation thinner comprises the oil-containing fraction that the vacuum greater than about 100mmHg vacuum is put on described extraction.

118. the method for claim 115, wherein said precipitation thinner comprises the oil-containing fraction that the vacuum greater than about 70mmHg vacuum is put on described extraction.

119. the method for claim 115, wherein said precipitation thinner comprises the oil-containing fraction that the vacuum greater than about 50mmHg vacuum is put on described extraction.

120. the method for claim 99, the step of the described oil-containing fraction of wherein said processing comprises deodorization.

121. the method for claim 120, wherein said deodorization are included in high temperature vacuum condition are put on the oil-containing fraction of described extraction, utilize steam to spray the oil-containing fraction of described extraction simultaneously.

122. the method for claim 121, wherein said high temperature are about 190 ℃ to about 220 ℃.

123. the method for claim 121, wherein said deodorization comprise the oil-containing fraction that the vacuum greater than about 25mmHg vacuum is put on described extraction.

124. the method for claim 121, wherein said deodorization comprise the oil-containing fraction that the vacuum greater than about 12mmHg vacuum is put on described extraction.

125. the method for claim 121, wherein said deodorization comprise the oil-containing fraction that the vacuum greater than about 6mmHg vacuum is put on described extraction.

126. the method for claim 99, wherein said oil product has the free fatty acid content less than about 0.5wt.%.

127. the method for claim 99, wherein said oil product has the free fatty acid content less than about 0.3wt.%.

128. the method for claim 99, wherein said oil product have the phosphorus value less than about 10ppm.

129. the method for claim 99, wherein said oil product have the phosphorus value less than about 5ppm.

130. the method for claim 99, wherein said oil product has the peroxide number less than about 2meq/kg.

131. the method for claim 99, wherein said oil product has the peroxide number less than about 1meq/kg.

132. the method for claim 99, wherein said oil product have the anisidine value less than about 5.

133. the method for claim 99, wherein said oil product have the anisidine value less than about 3.

134. the method for claim 99, wherein said oil product has the soap content less than about 5wt.%.

135. the method for claim 99, wherein said oil product has the soap content less than about 2.5wt.%.

136. the method for claim 99, wherein said oil product have the Fe concentration less than about 1ppm.

137. the method for claim 99, wherein said oil product have the Fe concentration of about 0.5ppm.

138. the method for claim 99, wherein said oil product have the Pb concentration less than about 1ppm.

139. the method for claim 99, wherein said oil product have the Pb concentration of about 0.2ppm.

140. the method for claim 99, wherein said oil product have the Hg concentration less than about 0.1ppm.

141. the method for claim 99, wherein said oil product have the Hg concentration of about 0.04ppm.

142. the method for claim 99, wherein said oil product have the Ni concentration less than about 0.1ppm.

143. the method for claim 99, wherein said oil product have the Ni concentration of about 0.01ppm.

144. the method for claim 99, wherein said oil product have the Cu concentration less than about 1ppm.

145. the method for claim 99, wherein said oil product have the Cu concentration of about 0.2ppm.

146. the method for claim 99, wherein said oil product experienced described blanching step before or after described treatment step.

147. the method for claim 99 also comprises described oil is separated into olein fraction and stearin fraction.

148. the method for claim 99, wherein said oil product are used for consuming for the people.

149. the method for claim 99, wherein said oil product is a solid at 20 ℃.

150. an oil product, the method by claim 99 prepares.

151. the microbial oil product that is used to consume, described oil product prepares by following steps: extract the oil-containing fraction from microbial biomass, wherein said oil-containing fraction comprises at least a LC-PUFA and is enough to the saturated fatty acid of the described oil-containing fraction of visual impact at least; Handle described fraction by the method for vacuum evaporation; Wherein said oil product does not experience the winterization step.

152. the microbial oil product of claim 151, wherein said oil product does not experience alkali refining process, cold filtration process or bleaching process.

153. the microbial oil product of claim 151, wherein said microbial biomass comes from and is selected from following microorganism: the microorganism that the microorganism that the microorganism that the microorganism that the microorganism that the microorganism that the microorganism of genus thraustochytrium, schizochytrium limacinum belong to, Althornia belong to, Aplanochytrium belong to, Japonochytrium belong to, Elina belong to, Crypthecodinium belong to, and their mixture.

154. coming from, the microbial oil product of claim 151, wherein said microbial biomass be selected from following microorganism: the microorganism that the microorganism that schizochytrium limacinum belongs to, Crypthecodinium belong to, and their mixture.

155. the microbial oil product of claim 151, wherein said oil product has the free fatty acid content less than about 0.5wt.%.

156. the microbial oil product of claim 151, wherein said oil product has the free fatty acid content less than about 0.3wt.%.

157. the microbial oil product of claim 151, wherein said oil product have the phosphorus value less than about 10ppm.

158. the microbial oil product of claim 151, wherein said oil product have the phosphorus value less than about 5ppm.

159. the microbial oil product of claim 151, wherein said oil product has the peroxide number less than about 2meq/kg.

160. the microbial oil product of claim 151, wherein said oil product has the peroxide number less than about 1meq/kg.

161. the microbial oil product of claim 151, wherein said oil product have the anisidine value less than about 5.

162. the microbial oil product of claim 151, wherein said oil product have the anisidine value less than about 3.

163. the microbial oil product of claim 151, wherein said oil product has the soap content less than about 5wt.%.

164. the microbial oil product of claim 151, wherein said oil product has the soap content less than about 2.5wt.%.

165. the microbial oil product of claim 151, wherein said oil product have the Fe concentration less than about 1ppm.

166. the microbial oil product of claim 151, wherein said oil product have the Fe concentration of about 0.5ppm.

167. the microbial oil product of claim 151, wherein said oil product have the Pb concentration less than about 1ppm.

168. the microbial oil product of claim 151, wherein said oil product have the Pb concentration of about 0.2ppm.

169. the microbial oil product of claim 151, wherein said oil product have the Hg concentration less than about 0.1ppm.

170. the microbial oil product of claim 151, wherein said oil product have the Hg concentration of about 0.04ppm.

171. the microbial oil product of claim 151, wherein said oil product have the Ni concentration less than about 0.1ppm.

172. the microbial oil product of claim 151, wherein said oil product have the Ni concentration of about 0.01ppm.

173. the microbial oil product of claim 151, wherein said oil product have the Cu concentration less than about 1ppm.

174. the microbial oil product of claim 151, wherein said oil product have the Cu concentration of about 0.2ppm.

175. the microbial oil product of claim 151, wherein said oil product is a solid at 20 ℃.

176. the microbial oil product of claim 151, wherein said oil product are used for consuming for the people.

177. a nutriment comprises the microbial oil product of claim 151.

178. a pharmaceutical products comprises the microbial oil product of claim 151.

179. the pharmaceutical products of claim 178, wherein said pharmaceutical products also comprises pharmaceutically acceptable excipient.

180. also comprising, the pharmaceutical products of claim 178, wherein said pharmaceutical products be selected from following medical active agent: the medicine of inhibin, antihypertensive, antidiabetic, anti-dull-witted medicine, antidepressants, antiadipositas drug, anorectic and raising memory and/or cognitive function.

181. a food comprises microbial oil product and the food or the liquid composition of claim 151.

182. the food of claim 182, wherein said food are selected from dough, batter, bake food, food liquid, semisolid food, be pressed into the food of piece, through pretreated meat, ice cream, frozen confectionery, ice yoghourt, magnificent husband's compound, salad flavoring, for the egg compound, become flavor snacks, special snacks, dried fruit snacks, meat flavour snacks, pork rind, the health food that is pressed into piece, rice/corn-dodger and confectionery snacks.

183. the microbial oil product that is used to consume, described oil product prepares by the method that may further comprise the steps:

A) extract the oil-containing fraction that comprises at least a LC-PUFA from microbial biomass; And

B) handle described fraction by the method for vacuum evaporation;

Wherein said oil product does not experience winterization step, alkali refining process, cold filtration process or bleaching process; And

Wherein said oil product has and is selected from following feature: free fatty acid content is less than about 0.5wt.%, the phosphorus value is less than about 10ppm, peroxide number is less than about 2meq/kg, anisidine value is less than about 5, and soap content is less than about 5wt.%, and Fe concentration is less than about 1ppm, Pb concentration is less than about 1ppm, Hg concentration is less than about 0.1ppm, Ni concentration less than about 0.1ppm and Cu concentration less than about 1ppm.

184. the microbial oil product of claim 183, wherein said oil product is a solid at 20 ℃.

185. a food comprises microbial oil product and the food or the liquid composition of claim 183.

186. a nutriment comprises the microbial oil product of claim 183.

187. a pharmaceutical products comprises the microbial oil product of claim 183.

188. the microbial oil product of claim 183, wherein said oil product are used for consuming for the people.

189. the method for the oil product that preparation is used to consume comprises:

A) extract the oil-containing fraction from microbial biomass, wherein said oil-containing fraction comprises at least a LC-PUFA; And

Wherein said oil product does not experience alkali refining process.

190. the method for claim 189, wherein said microorganism are the microorganism of Mortierella.

191. the method for claim 189, wherein said oil-containing fraction comprises arachidonic acid.

192. the method for claim 189, wherein said oil product is a solid at 20 ℃.

193. an oil product, the method by claim 189 prepares.

194. the oil product of a mixing comprises the oil product of claim 151 and the oil product of claim 193.

195. the oil product of the mixing of claim 194, the microbial biomass that wherein prepares the oil product of claim 151 comes from and is selected from following microorganism: the microorganism that the microorganism that the microorganism that the microorganism that the microorganism that the microorganism that the microorganism of genus thraustochytrium, schizochytrium limacinum belong to, Althornia belong to, Aplanochytrium belong to, Japonochytrium belong to, Elina belong to, Crypthecodinium belong to, and their mixture.

196. the oil product of the mixing of claim 194, wherein said microorganism are selected from the microorganism of schizochytrium limacinum genus, the microorganism that Crypthecodinium belongs to, and their mixture.

197. the oil product of the mixing of claim 194, the microbial biomass that wherein prepares the oil product of claim 193 comes from the microorganism of Mortierella.

198. the oil product of the mixing of claim 194, wherein said product comprises DHA and arachidonic acid.

199. preparation comprises the fluid oil fraction of LC PUFA and comprises the method for the solid-state fat products of LC PUFA, comprise: rough microbial oil is separated into oil product and solid-state fat products, and wherein said solid-state fat products has the LC PUFA content at least about 20% weight.

200. the method for claim 199, wherein said solid-state fat products has the DHA content at least about 20% weight.

201. the method for claim 199, wherein said separating step comprise described rough microbial oil is contacted with adsorbent, also comprises:

A) isolate the material of described adsorbent and absorption from described rough microbial oil;

B) material from described adsorbent and absorption reclaims described solid-state fat products.