CN104783175B

CN104783175B - Novel microalgae food composition

Info

Publication number: CN104783175B
Application number: CN201510071275.9A
Authority: CN
Inventors: G·布鲁克斯; S·富兰克林; J·阿维拉; S·M·戴克尔; E·巴利乌; W·莱基特斯基; J·皮朔基; D·兹达尼斯; L·M·诺里斯
Original assignee: Terraria Holdings
Current assignee: Kirbyn Biotech Corp
Priority date: 2009-04-14
Filing date: 2010-04-14
Publication date: 2017-08-22
Anticipated expiration: 2030-04-14
Also published as: CN104783175A

Abstract

The invention provides novel microalgae food composition, it includes having been processed into the microalgae biomass for thin slice, powder and powder, a kind of food ingredients composition for the egg products and algae powder for including drying is especially provided, the algae powder is the homogenate of microalgae biomass, the microalgae biomass mainly or entirely contains the dissolving cell in powder type, include by dry weight at least 16% triglyceride oil, for preparing a kind of food article by adding a kind of liquid and optional other comestible compositions.Microalgae biomass saturated fat levels of the invention are low, single unsaturated triglyceride oil content is high, and can be the good sources of fiber.

Description

Novel microalgae food composition

The application is the PCT application on April 14th, 2010 " novel microalgae food composition " submit, entitled PCT/US2010/031088 divisional application, the date that the PCT application enters National Phase in China is December 13 in 2011 Day, Application No. 201080026237.7.

The cross reference of related application

The U.S. Provisional Patent Application No. 61/ that the application on April 14th, 2009 is submitted by 35U.S.C.119 (e) requirements 169,271st, U.S. that on April 27th, 2009 submits U.S. Provisional Patent Application No. 61/173,166, September in 2009 are submitted on the 25th The U.S. Provisional Patent Application No. 61/299,250 that state's Provisional Patent Application No. 61/246,070 and on January 28th, 2010 submit Rights and interests.The part for the international application no PCT/US2009/060692 that the application or on October 14th, 2009 submit after Continuous application, U.S. Provisional Patent Application No. 61/173,166 that it on April 27th, 2009 is submitted by 35U.S.C.119 (e) requirements, The rights and interests for the U.S. Provisional Patent Application No. 61/246,070 that September in 2009 is submitted on the 25th.The application or on January 8th, 2010 The U.S. Application No. 12/684,884,12/684,885,12/684,886,12/684,887,12/684,888,12/ of submission 684,889,12/684,891,12/684,892,12/684,893 and 12/684, a 894 part continuation application, and And each of which is the part continuation Shen for the U.S. Provisional Patent Application No. 12/579,091 submitted on October 14th, 2009 Please, the U.S. Provisional Patent Application No. 61/173,166,2009 that it on April 27th, 2009 is submitted by 35U.S.C.119 (e) requirements The rights and interests for the U.S. Provisional Patent Application No. 61/246,070 that on September is submitted for 25.For all purposes by quoting these Each of application is combined herein with its full text.

Quote sequence table

The application is included in an entitled 026172-004150PC sequence table of creation on April 14th, 2010 and wrapped A sequence table in text containing 22803 bytes.The material that will be contained in hereby in this article this document by quote into Row is combined.

Technical field

The field of the invention is the nutrition of microbiology, food preparation and the mankind and animal.

Background technology

With the continuous growth of population, extra food source is had increasing need for, particularly can inexpensively be produced but nutrition Abundant food source.In addition, at least in most of developed country, many staple foods also rely on meat at present, this is greatly Promote the discharge of greenhouse gases, it is therefore desirable to which production is same tasty, nutritious but former to the less new food of environmental hazard Material.

The growth of algae only needs to " water and sunlight ", therefore it is expected to as potential food source always.Although some Algae (mainly sea grass) provides important food material for the consumption of the mankind really, but is used as using algae the hope of food material Prestige does not become a reality also.The algae powder being made with the algae of photosynthetic growth in pond out of doors or bioreactor is can business Purchase, but its color is dark green (coming from chlorophyll), and with strong, undesirable taste.Prepared when by these algae powder When among to food article or as nutritional supplement, it is visually unattractive that they produce food article or nutritional supplement Green color, and with unpleasant fish raw meat or sea grass taste.

There is several algae to be used in food material at present, majority is large-scale algae, such as sea-tangle, seaweed (Porphyra, for sea Tongue piece), dulse (red algae) and extra large lettuce (sea lettuce).Microalgae (such as spirulina (blunt item arthrospira)) is the business in open pond Change (photosynthetic) of plantation, as nutritional supplement, or marginally mix in smoothie or fruit drink (typically less than 0.5%w/ w).Prevalence is used as nutritional supplement to other microalgaes (including some species of chlorella) in Asian countries.

In addition to these products, the algae oil with high docosahexaenoic acid (DHA) content is used in infant formula Composition.DHA is height polyunsaturated oil.DHA has anti-inflammatory property, is well-known tonic and is prepared used in food material In additive.However, DHA is not appropriate for prepared food, because it thermally treated can be aoxidized.In addition, even in room temperature and depositing In the case of antioxidant, DHA is also unstable when exposed to oxygen.DHA oxidation produces fishy taste and unpleasant Smell.

Present need exist for producing food material on a large scale by algae using the method for Cheap highly effective, particularly production can Mouth and nutritious food material.The present invention can meet these and other demand.

The content of the invention

The present invention includes the composition of multiple genus and species from edible microalgae and the powder for coming from microalgae of strain.At this The single unsaturated triglyceride oil of primary of the microalgae used in invention without Algae toxins and containing varying level.Draped over one's shoulders at this The powder of dew be configured to oxidation-stabilized, homogeneous and micronizing, flow freely can blended powder, mixing food ingredients, And combination therein.Disclosed powder also forms the emulsion of homeostasis in the slurry herein, with manageable viscosity. Further disclose the inventive process prepared powder and be bonded to enter food composition.Present invention additionally comprises with significant Digestible protein and unique dietary fiber content and the powder of associated water combination, systematism and healthy oil transport attributes.Also Disclose the novel method that oil ＆ fat is substituted using the powder of the present invention.Can be from edible and inedible heterotrophism Fermentation raw material, including cornstarch, sugarcane, glycerine and depolymerizing cellulose manufacture powder of the invention.

In a first aspect, the invention provides a kind of microalgae powder, it is homogenized including microalgae biomass, the microalgae biomass Mainly or entirely contain the dissolving cell in powder type, this powder includes by dry weight at least 16% triglycerides Oil.In some embodiments, the particle mean size of the particle in powder is less than 100 μm.In some embodiments, in powder The particle mean size of particle is 1-15 μm.In one embodiment, the powder is by the way that microalgae biomass micronizing is formed A kind of emulsion and make the emulsion dry formed by.In some cases, the microalgae powder have by weight 10% or less, Or 5% or less moisture.In some cases, the biomass includes glycerine by dry weight between 45% and 70% Three ester oils.In some embodiments, 60% to 75% triglyceride oil be it is a kind of in glyceride form 18:1 lipid. In one embodiment, the triglyceride oil is less than 2%14:0th, 13% to 16%16:0th, 1% to 4%18:0th, 64% to 70%18:1st, 10% to 16%18:2nd, 0.5% to 2.5%18:3 and carbon chain lengths less than 2% be 20 or longer oil. In some embodiments, the biomass includes carbohydrate by dry weight between 25% and 40%.In certain situation Under, the carbohydrate ingredient of the biomass is dietary fiber by dry weight between 25% and 35% and in 2% and 8% Between free sugar (including sucrose).In one embodiment, the monose of the dietary fiber component of the biomass, which is constituted, is 0.1% to 4% arabinose, 5% to 15% mannose, 15% to 35% galactolipin and 50% to 70% Glucose.In some cases, biomass has the total carotinoid between 20 to 115 μ g/g, includes 20-70 μ g/g leaf Flavine.In some cases, the chlorophyll content of the biomass is less than 2ppm.In some cases, the biomass has 1- 8mg/100g total tocopherols, include 2-6mg/100g alpha tocopherol.In some cases, the biomass has 0.05- 0.30mg/g total tocotrienols, includes 0.10-0.25mg/g α tocotrienols.

In some embodiments, the microalgae powder end is in a kind of form of food ingredients composition, the wherein microalgae powder End is combined with the other comestible compositions of one or more, these comestible compositions be a kind of cereal, fruit, vegetables, protein, Herbaceous plant, spices or it is one or more be used to preparing salad-dressing, egg products, baked goods, bread, pasta, Sauce, soup, beverage, frozen dessert, butter smear food (spread).In some cases, the microalgae powder lacks visible Oil.In some cases, the microalgae powder further comprises glidant.In one embodiment, the microalgae powder further comprises Antioxidant.

In the different embodiments of the present invention, the biomass is derived from a single strain of microalgae.In some embodiment party In case, the biomass sources are in the algae of a kind of Chlorella.In one embodiment, the algae is Chlorella protothecoides (Chlorella protothecoides).In some cases, the biomass is derived from having compared with from its derivative strain A kind of algae of the reduced colour mutant of pigment formation.In some embodiments, the algal biomass is derived from Heterotrophic culture Algae.In some embodiments, the algal biomass is derived from what is cultivated and process under the conditions of Good Manufacture Practice (GMP) Algae.

In second aspect, the invention provides a kind of food ingredients composition, it include following component or by combine with Lower composition and formed：(a) at least 0.5%w/w microalgae powder, the wherein microalgae powder are mainly or entirely to contain to be in powder type Dissolving microalgae cell, this powder includes by weight at least 16% triglyceride oil, and (b) at least one other Comestible composition, wherein the food ingredients composition can be become by the way that a kind of liquid is added into the food ingredients composition Into a kind of food article of recovery.In one embodiment, the food ingredients composition is dry pasta.One In the case of a little, the food ingredients composition can be changed to a kind of food article of recovery by adding liquid after baking. In one embodiment, the food article of the recovery is liquid food finished product.In some cases, the food ingredients composition can To be changed to the food article restored by a kind of technique, the technique includes making the product of regeneration be subjected to shearing force.At some In embodiment, the particle mean size of the particle of the microalgae biomass in liquid food products is between 1 and 15 μm.At one In embodiment, the food article of the recovery is a kind of emulsion.In some embodiments, the food article of the recovery is salad Baste, soup, sauce, beverage, butter smear food (spread).

In some cases, the food article of recovery of the invention except from microalgae biomass oil in addition to do not include oil or Fat.In some embodiments, the value of microalgae powder is same with the food article of the recovery in the food article of the recovery 0.25-1 times of oily and/or fat weight in the conventional food article of sample type.In some cases, of microalgae biomass The particle mean size of grain is less than 100 μm.In some embodiments, the particle mean size of the particle of microalgae biomass is 1-15 μm. In some embodiments, the food composition has by weight 10% or less or 5% or less moisture.One In the case of a little, the microalgae biomass includes triglyceride oil by dry weight between 45% and 65%.In some embodiments In, 60% to 75% triglyceride oil be it is a kind of in glyceride form 18:1 lipid.In one embodiment, this is sweet Oily three ester oils are less than 2%14:0th, 13% to 16%16:0th, 1% to 4%18:0th, 64% to 70%18:1st, 10% to 16% 18:2nd, 0.5% to 2.5%18:3 and carbon chain lengths less than 2% be 20 or longer oil.In some cases, the biology Matter is carbohydrate by dry weight between 25% to 40%.In some cases, the carbon hydrate of the microalgae biomass Thing component is the free sugar (including sucrose) between the dietary fiber and 2% to 8% between by dry weight 25% to 35%. In one embodiment, the dietary fiber component of the microalgae biomass monose constitute be 0.1% to 4% arabinose, 5% to 15% mannose, 15% to 35% galactolipin and 50% to 70% glucose.In some cases, microalgae Biomass has the total carotinoid between 20 to 115 μ g/g, includes 20-70 μ g/g lutein.In some cases, should Microalgae biomass includes the chlorophyll less than 2ppm.In some embodiments, the microalgae biomass has 1-8mg/100g's Total tocopherols, include 2-6mg/100g alpha tocopherol.In one embodiment, the microalgae biomass includes 0.05- 0.30mg/g total tocotrienols, includes 0.10-0.25mg/g α tocotrienols.

In the third aspect, the invention provides a kind of method for manufacturing microalgae powder, it includes (a) and provided containing by dry weight The microalgae cell of at least 16% triglyceride oil, (b) makes these clasmatosises and granularity is reduced to produce aqueous homogenate, And (c) makes the homogenate drying to produce including at least microalgae powder of 16% triglyceride oil by dry weight.In an embodiment party In case, this method further comprises separating these microalgae cells from culture medium before these clasmatosises are made.In some feelings Under condition, crushed using pressure breaking instrument, French cell press or ball mill.In some cases, using freeze-drying Device, drum dryer, flash dryer, spray dryer or chamber dryer are dried.

In some embodiments, this method is to use the oily microalgae contained by dry weight between 50% and 65% thin Born of the same parents are carried out.In some cases, this method further comprises that any point during technique adds a kind of glidant.One In individual embodiment, the particle mean size of the particle in powder is less than 100 μm.In one embodiment, the particle mean size of powder It is between 1 and 15 μm.In some cases, the moisture that the powder has by weight 10% or less or 5% or less contains Amount.In some cases, 50% to 75% oil be it is a kind of in glyceride form 18:1 lipid.In an embodiment In, the oil is less than 2%14:0th, 13% to 16%16:0th, 1% to 4%18:0th, 64% to 70%18:1st, 10% to 16%18: 2nd, 0.5% to 2.5%18:3 and carbon chain lengths less than 2% be 20 or longer oil.

In some cases, this method is carried out using the microalgae cell of a single strain of microalgae.In some realities Apply in scheme, these cells are one kind of Chlorella.In one embodiment, these cells are Chlorella protothecoides.One In the case of a little, these cells belong to a kind of colour mutant, and this mutant strain has reduced color compared with from its derivative strain Element is formed.In some cases, these cells are from Heterotrophic culture.In some embodiments, these cells are made good Quite crush and dry under the conditions of production specification (GMP).

In fourth aspect, it is based on the invention provides the method that one manufactures food article from microalgae powder, including (a) normal The amount of oil, fat or egg in the food article of rule form, it is determined that the amount for the microalgae powder that will be included in the food article, wherein The microalgae powder is the homogenate of microalgae biomass, the microalgae biomass mainly or entirely contain comprising by dry weight at least 16% it is sweet The dissolving cell in powder type of oily three ester oils；And (b) is by the microalgae powder of the amount and one or more comestible compositions Combination (and less than the amount for the oil, fat or egg being present in the food article of conventionally form), to be somebody's turn to do from microalgae powder formation Food article.In some cases, in addition to the microalgae oil for belonging to the biomass, the food article contains to be less than by weight 25% oily or fat.In some cases, in addition to the microalgae oil for belonging to the biomass, the food system from microalgae powder Product include the oily or fat by weight less than 10%.In one embodiment, except the microalgae for belonging to the biomass is oily Outside, the food article from microalgae powder not oil-containing or fat.In some cases, the food article from microalgae powder does not include Egg.In some cases, the food article is free of the oil in addition to the microalgae oil for belonging to the biomass.

The present invention further comprises the beverage of novelty and for manufacturing their raw material, and this kind of beverage and raw material contain There is the microalgae not of the same race with different component.Attribute for the microalgae biomass of the present invention includes the material for providing nutrition Material, such as carrotene, dietary fiber, tocotrienols and tocopherol and different lipid compositions, special low-level Saturation lipid.Attribute for the microalgae biomass of the present invention includes structure attribute, such as with alternative dairy products (such as soya-bean milk With Rice ＆ peanut milk) compare improved mouthfeel.The novel beverage provides the delivering for high nutrition material found in microalgae System.The invention provides a kind of finished beverage of the new category based on microalgae (as freezing and shelf-stable liquid and Emulsion) together with the combination of the characteristic by being used to increase current beverage as composition comprising the novel material based on microalgae Thing.

At the 5th aspect, the invention provides a kind of beverage including microalgae biomass and edible liquid, the microalgae Biomass comprising at least 16% triglyceride oil by dry weight and/or at least 40% in full cell or mainly or entirely contain There is the dried protein of the form of the homogenate of dissolving cell.In some cases, the beverage is by scattered microalgae biomass and can Formed by edible liquid.In some cases, the microalgae biomass is the form in micronizing homogenate.In an embodiment party In case, the particle mean size of the particle in homogenate is less than 100 μm.In one embodiment, the particle mean size of the particle in homogenate It is 1-15 μm.

In some cases, the biomass by mass spectral analysis without the Algae toxins that can be detected.In one embodiment, The beverage is pasteurizing.In some embodiments, the beverage further comprises a kind of exogenous proteins source and/or breast Sugar.In one embodiment, the exogenous proteins source is lactalbumin.In some cases, the beverage is free of lactose.One In the case of a little, the edible liquid is soya-bean milk, Rice ＆ peanut milk or almond milk.

In some embodiments, the beverage is to be selected from the group, and the group is made up of the following：Milk, fruit juice, smoothie, Nutritious drink, eggnog and meal replacement beverage.In some cases, the microalgae biomass is by dry weight 45% to 75% Triglyceride oil.In one embodiment, by weight at least 50% triglyceride oil is single unsaturated oils.At one In embodiment, by weight at least 50% triglyceride oil is 18:1 lipid and in the form of glyceride and by comprising. In some cases, the triglyceride oil by weight less than 5% is docosahexaenoic acid (DHA) (22:6).In some feelings Under condition, 60% to 75% triglyceride oil be it is a kind of in glyceride form 18:1 lipid.In one embodiment, should Triglyceride oil is less than 2%14:0th, 13% to 16%16:0th, 1% to 4%18:0th, 64% to 70%18:1st, 10% to 16% 18:2nd, 0.5% to 2.5%18:3 and carbon chain lengths less than 2% be 20 or longer oil.

In some embodiments, the biomass is carbohydrate by dry weight between 25% and 40%.One In the case of a little, the carbohydrate ingredient of the biomass is dietary fiber and 2% by dry weight between 25% and 35% With the free sugar (including sucrose) between 8%.In one embodiment, the monose of the dietary fiber component of the biomass is constituted Be 0.1% to 4% arabinose, 5% to 15% mannose, 15% to 35% galactolipin and 50% to 70% Glucose.In some cases, the biomass has the total carotinoid between 20 to 115 μ g/g, including 20-70 μ g/g Lutein.In one embodiment, the chlorophyll content of the biomass is less than 2ppm.In some cases, the biomass has There are 1-8mg/100g total tocopherols, include 2-6mg/100g alpha tocopherol.In some cases, the biomass has 0.05-0.30mg/g total tocotrienols, includes 0.10-0.25mg/g α tocotrienols.

In some cases, microalgae of the biomass from heterotrophic growth.In some cases, the biomass is well being given birth to Manufactured under production normality condition.In some cases, the microalgae biomass is derived from a single strain of microalgae.In some implementations In scheme, the microalgae is a kind of Chlorella.In one embodiment, the microalgae is Chlorella protothecoides (Chlorella Protothecoides a strain).In some cases, the biomass is derived from having pigment compared with from its derivative strain Form a kind of algae of reduced colour mutant.In one embodiment, the microalgae was preserved on October 13rd, 2009 The Chlorella protothecoides 33-55 of American type culture collection is deposited under title PTA-10397.In an embodiment In, the microalgae is to be deposited in American type culture collection in the case where preserving title PTA-10396 on October 13rd, 2009 Chlorella protothecoides 25-32.

At the 6th aspect, the method for beverage is manufactured the invention provides a kind of, including full cell sheet or powder will be in Form microalgae biomass or micronizing homogenate in powder type with least 25% triglycerides oil content can with one kind Edible liquid combines to form a kind of beverage.In some cases, the microalgae biomass first with a kind of second edible liquid Combination is to form a kind of slurry, and then the slurry combines to form beverage with the edible liquid.In some cases, should Method further comprises adding a kind of exogenous proteins source and/or lactose forming the beverage.In one embodiment, this is outer Source protein matter source is lactalbumin.In some cases, this method further comprises the beverage carrying out pasteurizing.One In the case of a little, the microalgae biomass is combined to form a kind of dispersion of stabilization with edible liquid.

In different embodiments, the beverage manufactured by the method for the present invention is to be selected from the group, and the group is by following Item composition：Milk, fruit juice, smoothie, nutritious drink and meal replacement beverage.In some cases, the edible liquid is beans Slurry, Rice ＆ peanut milk or almond milk.

At the 7th aspect, by dry weight at least 16% is included the invention provides a kind of food article of fermentation, including (a) Triglyceride oil and/or at least 40% homogenate in full cell or mainly or entirely containing dissolving cell form it is dry Protein, a kind of (b) edible liquid, and (c) are a kind of suitable for the microorganism of the work used in food article.In some feelings Under condition, the microorganism of the work is a kind of bacterial cultures.In one embodiment, the edible liquid is milk.In a reality Apply in scheme, the milk comes from animal.In one embodiment, the milk comes from non-animal.In some cases, the fermentation Food article be Yoghourt.In some cases, the Yoghourt is the form in liquid beverage.

The present invention further comprises the baked goods containing microalgae, and compared to the homotype product previously existed, it has novelty Characteristic.It there is disclosed herein and prepare and manufacture these foods to deliver reduced fat, the cholesterol of reduction and increased fibre The method of dimension hplc.Different embodiments includes the favourable of egg, butter, animal tallow and the saturated oils of elimination or reduction In the oil containing microalgae biomass and oils of health, including manufacture is with the homotype product more low-calorie than previously having existed Food.Additionally provide production be used for manufacture novelty treated baked goods and intermediate product (such as cake and bead mix Compound) raw material method.

In eighth aspect, the invention provides one kind by bakeing microalgae biomass and a kind of edible liquid and at least one Food article formed by the mixture of other comestible compositions is planted, the microalgae biomass has by weight at least 16% Triglycerides oil content, the shape of the homogenate in full cell sheet or full cell powder or mainly or entirely containing dissolving cell Formula.In some cases, the microalgae biomass is in the form of microalgae powder, and it is mainly or entirely containing micro- in powdered The homogenate of the dissolving cell of algal biomass.In some cases, the microalgae powder is the homogenate of the micronizing of microalgae biomass.One In the case of a little, the microalgae biomass is the form of the slurry in homogenate.

In some embodiments, by the mass spectral analysis biomass without the Algae toxins that can be detected.In some cases, The food article has the water activity (Aw) between 0.3 and 0.95.In some cases, it is conventional with a kind of other identical Food article compare, the food article has at least 1.5 times higher fiber contents.In some cases, the food article It is to be selected from the group, the group is made up of the following：Brownie, cookies, cake and pie product, crispbread, bread and Snack chip.In some cases, the bread is pizza crust, bread stick, cream roll or biscuit.In some cases, this is micro- Algal biomass be by dry weight from 45% to 75% triglyceride oil.In some cases, by weight at least 50% it is sweet Oily three ester oils are single unsaturated oils.In one embodiment, by weight at least 50% triglyceride oil is 18:1 lipid And in the form of glyceride and by comprising.In some cases, the triglyceride oil by weight less than 5% is 22 carbon Acid (DHA) (22:6).In some cases, 60% to 75% triglyceride oil is a kind of in glyceride form 18:1 lipid.In one embodiment, the triglyceride oil is less than 2%14:0th, 13% to 16%16:0th, 1% to 4% 18:0th, 64% to 70%18:1st, 10% to 16%18:2nd, 0.5% to 2.5%18:3 and carbon chain lengths less than 2% be 20 Or longer oil.

In some cases, the biomass be by dry weight 25% and with 40% between carbohydrate.At some In the case of, the carbohydrate ingredient of the biomass is dietary fiber by dry weight between 25% and 35% and 2% With the free sugar (including sucrose) between 8%.In one embodiment, the monose of the dietary fiber component of the biomass is constituted Be 0.1% to 3% arabinose, 5% to 15% mannose, 15% to 35% galactolipin and 50% to 70% Glucose.In some cases, biomass has the total carotinoid between 20 to 115 μ g/g, includes 20-70 μ g/g leaf Flavine.In one embodiment, the chlorophyll content of the biomass is less than 2ppm.In one embodiment, the biomass Total tocopherols with 1-8mg/100g, include 2-6mg/100g alpha tocopherol.In some cases, the biomass has 0.05-0.30mg/g total tocotrienols, includes 0.10-0.25mg/g α tocotrienols.

In some cases, microalgae of the biomass from heterotrophic growth.In some cases, the biomass is well being given birth to Manufactured under production normality condition.In some embodiments, the microalgae biomass comes from the microalgae of a kind of Chlorella. In one embodiment, the microalgae is a strain of Chlorella protothecoides (Chlorella protothecoides).At some In the case of, the microalgae biomass is derived from the one kind for the colour mutant for having pigment formation reduced compared with from its derivative strain Algae.In one embodiment, the microalgae is to be deposited in U.S.'s allusion quotation in the case where preserving title PTA-10397 on October 13rd, 2009 The Chlorella protothecoides 33-55 of type culture collection.In one embodiment, the microalgae be on October 13rd, 2009 by Preserve the Chlorella protothecoides 25-32 that American type culture collection is deposited under title PTA-10396.

At the 9th aspect, the invention provides a kind of food ingredients composition, it includes microalgae biomass and at least one Other comestible compositions, the microalgae biomass is in full cell sheet or full cell powder or mainly or entirely thin containing dissolving The form of the homogenate of born of the same parents, with by weight at least 16% triglycerides oil content, the wherein food ingredients can be by adding Liquid feeding body is into the food ingredients composition and bakees and is converted into the food article of recovery.In some cases, the biology The triglycerides oil content that matter has for by dry weight 45% to 70% triglyceride oil.In some cases, the biology Matter includes by dry weight at least 40% protein, and the protein includes at least 60% digestible crude protein matter.

At the tenth aspect, the invention provides a kind of method for manufacturing baked product, including by microalgae biomass with it is a kind of Edible liquid and at least one other comestible composition combinations, and the mixture is bakeed, the microalgae biomass, which has, to be pressed The triglycerides oil content of weight meter at least 25%, the micro mist in full cell sheet or full cell powder or in powder type Change homogenate.In some cases, the baking goodses are Brownie, cookies, cake, bread, pizza crust, bread stick, wafer Dry, biscuit, pie crust or snack chip.In some cases, the microalgae biomass not with a kind of edible liquid or its His comestible composition (mainly fat, oil or egg) combination.

The tenth on the one hand, the invention provides a kind of food article, it includes microalgae biomass and a kind of edible liquid Body and a kind of powder, the microalgae biomass have by weight at least 10% triglycerides oil content, in full cell sheet Or full cell powder or the form of the homogenate mainly or entirely containing dissolving cell.In some cases, the food article enters one Step includes leavening agent.In one embodiment, the leavening agent is chemical leavening agent.In one embodiment, the leavening agent It is biological leavening agent.In some cases, the microalgae biomass includes triglycerides by dry weight between 45% and 70% Oil.In some cases, the microalgae biomass includes at least 40% protein.

The present invention further comprises the food for including the microalgae biomass with high-level lipid.The example of food includes sand Department, flavouring, smear food, mayonnaise and other contain edible materials of microalgae, wherein these edible materials are conventional It is upper associated with the delivering of saturated fat and oil.Invention further provides the food containing microalgae, these foods with it is same The conventional food of sample type compares the calorie load with reduction, and in various embodiments, these novel foods With the similar or identical organoleptic attribute to the full-cream fat pattern of these foods.Additionally provide preparation and manufacture these novelty foods The method of thing and for manufacturing for the method for the intermediate product based on microalgae that manufactures them.These novel foods are with Between product can use existing fermentation and food processor appliances and manufacture, and can be with apparatus desired structure and sense organ The more healthy food for coming from microalgae of characteristic replaces existing food article.

At the 12nd aspect, the invention provides a kind of food or food ingredients composition, it is included by weight at least The homogenate of 10% microalgae biomass, the microalgae biomass is mainly or entirely containing dissolving cell, including by dry weight at least 16% triglyceride oil emulsified in edible liquid.In some cases, said composition be baste, mayonnaise, soup, Or flavouring.In some cases, said composition is free of the oil ＆ fat in addition to the oil in the microalgae biomass.One In the case of a little, in addition to the oil for belonging to biomass, said composition contains the oily or fat by weight less than 25%.At some In the case of, in addition to the oil for belonging to biomass, said composition contains the oily or fat by weight less than 10%.In a reality Apply in scheme, said composition is oil-in-water emulsion.In one embodiment, said composition is water-in-oil emulsion.

In some cases, by the mass spectral analysis biomass without the Algae toxins of level can be detected.In some cases, The microalgae biomass be by dry weight 45% to 75% triglyceride oil.In some cases, by weight at least 50% Triglyceride oil is single unsaturated oils.In some cases, by weight at least 50% triglyceride oil is 18:1 lipid is simultaneously And in the form of glyceride and by comprising.In some cases, the triglyceride oil by weight less than 5% is 22 carbon six Olefin(e) acid (DHA) (22:6).In some cases, 60% to 75% triglyceride oil be it is a kind of in glyceride form 18:1 Lipid.In one embodiment, the triglyceride oil is less than 2%14:0th, 13% to 16%16:0th, 1% to 4%18:0、 64% to 70%18:1st, 10% to 16%18:2nd, 0.5% to 2.5%18:3 and carbon chain lengths less than 2% be 20 or more Long oil.

In some cases, the biomass includes carbohydrate by dry weight between 25% and 40%.At some In the case of, the carbohydrate ingredient of the biomass be dietary fiber between by dry weight 25% to 35% and 2% to Free sugar (including sucrose) between 8%.In one embodiment, the monose of the dietary fiber component of the biomass, which is constituted, is 0.1% to 4% arabinose, 5% to 15% mannose, 15% to 35% galactolipin and 50% to 70% Glucose.In some cases, the biomass has the total carotinoid between 20 to 115 μ g/g, including 20-70 μ g/g Lutein.In one embodiment, the chlorophyll content of the biomass is less than 2ppm.In some cases, the biomass has There are 1-8mg/100g total tocopherols, include 2-6mg/100g alpha tocopherol.In some embodiments, the biomass has There is 0.05-0.30mg/g total tocotrienols, include 0.10-0.25mg/g α tocotrienols.

In some cases, microalgae of the biomass from heterotrophic growth.In some cases, the biomass is good Manufactured under production normality condition.In some cases, the microalgae biomass is derived from Chlorella, Prototheca (genus Prototheca) or class Chlorella (genus Parachlorella) a kind microalgae.In some embodiments In, the microalgae is a kind of Chlorella.In one embodiment, the microalgae is Chlorella protothecoides (Chlorella Protothecoides a strain).In one embodiment, the microalgae is to preserve title on October 13rd, 2009 The Chlorella protothecoides 33-55 of American type culture collection is deposited under PTA-10397.In one embodiment, should Microalgae is that the protoconch of American type culture collection is deposited in the case where preserving title PTA-10396 on October 13rd, 2009 Chlorella 25-32.

At the 13rd aspect, the invention provides one kind by slurry formed by scattered algae powder, it is microalgae biomass Homogenate, the microalgae biomass is mainly or entirely containing dissolving cell, including at least 16% place in aqueous by dry weight In the triglyceride oil of powder type, the wherein algae powder constitutes by weight the 10% to 50% of the slurry.In certain situation Under, the oil content that the biomass has for by dry weight 5% to 55% triglyceride oil.In some cases, the biology Matter includes by dry weight at least 40% protein, and the protein includes at least 60% digestible crude protein matter.

In fourteenth aspect, it is based on the invention provides the method that manufacture includes microalgae biomass food article, including (a) The amount of oil, fat or egg in the food article of conventionally form, it is determined that the microalgae biomass in the food article will be included in Amount, wherein the microalgae biomass comprising by dry weight at least 16% triglyceride oil；And (b) is biological by the microalgae of the amount Matter combine with one or more comestible compositions (and less than the oil, fatty or egg being present in the food article of conventionally form Amount) to form the food article that this includes microalgae biomass.In some cases, except belonging to the microalgae oil of the biomass Outside, the food article contains the oily or fat by weight less than 10%.In some cases, except belonging to the micro- of the biomass Outside algae oil, the food article comprising microalgae powder is without the main food ingredients being made up of oil or fat.In some cases, wrap Food article containing microalgae powder is free of egg.In some embodiments, the food article is free of except belonging to the micro- of the biomass Oil beyond algae oil.In some cases, the amount of microalgae biomass be in conventional formulation by weight or stereometer 25% to The oily or fat of 100% volume.

At the 15th aspect, the invention provides a kind of method for manufacturing low-fat diet, including it will include by dry weight extremely The algal biomass of few 16% triglyceride oil is combined with one or more other comestible compositions, and wherein at least one is edible Exhausted with composition in terms of natural fat or oil.In some cases, the edible exhausted in terms of natural fat or oil Composition is a kind of albumen.In some cases, the comestible composition exhausted in terms of natural fat or oil is one kind in fat The dairy products that aspect has exhausted.In one embodiment, the dairy products are reduction or non-fat milk.

At the 16th aspect, the invention provides a kind of method for manufacturing low-fat diet, it includes will be comprising by dry weight The algal biomass of at least 16% triglyceride oil is combined to form a kind of low fat with one or more other comestible compositions Food article, wherein the low-fat diet product have the oily or fat no more than 10% in addition to microalgae oil.In some cases, The algal biomass one or more comestible compositions in combination do not include the main composition being made up of oil, fat or egg.

The present invention further comprises the composition relevant with the generation of egg based food article and method, and wherein these are produced Product contain the different raw material manufactured from various forms of microalgaes.Some forms include high-caliber single unsaturated oils, meals Fiber, carotenoid and digestible crude protein matter.It there is provided herein the extended period for being stored in the egg products of hydration The method and composition of food stabilization is improved between time limit at elevated temperatures.There is provided the different genus and species produced by heterotrophism The material for coming from microalgae being homogenized as drying or being hydrated manufactured with the microalgae of strain.Reduced in the egg products of the present invention full With fat and the w/w level of cholesterol, while adding dietary fiber.There is provided egg liquid or powdered egg and liquid algae or dry The admixture of dry algae, additionally provides manufacture and the method for preparing the admixture.Additionally provide for manufacturing very low cholesterol Egg products albumen and the unique combination of microalgae.In some embodiments, the dietary fiber comprising microalgae biomass is passed through And other humidity retention performances and by the texture characteristic change of powdered egg into the texture feature more like egg liquid.

At the 17th aspect, the invention provides a kind of food ingredients composition including drying egg products and algae powder, algae Powder is the homogenate of microalgae biomass, and the microalgae biomass mainly or entirely contains the dissolving cell in powder type, including by The triglyceride oil of dry weight meter at least 16%, for preparing food by adding liquid and optional other comestible compositions Tetramune.In one embodiment, the egg products of the drying are dry shell eggs.In one embodiment, the drying Egg products are dry albumen.In one embodiment, the egg products of the drying are dry yolk.In some cases, The food ingredients composition is powdery egg products or pancake or waffle powder.

In some cases, the algae powder is to form a kind of emulsion by the way that microalgae biomass is micronized and do the emulsion Formed by dry.In one embodiment, the particle mean size of the particle in algae powder is less than 100 μm.In one embodiment, The particle mean size of particle in algae powder is 1-15 μm.In some cases, the biomass is manufactured under the conditions of Good Manufacture Practice 's.In some cases, by the mass spectral analysis biomass without the Algae toxins that can be detected.

In some cases, the microalgae biomass be by dry weight 45% to 75% triglyceride oil.In a reality Apply in scheme, by weight at least 50% triglyceride oil is single unsaturated oils.In one embodiment, by weight extremely Few 50% triglyceride oil is 18:1 lipid and in the form of glyceride and by comprising.In one embodiment, by weight Triglyceride oil of the gauge less than 5% is docosahexaenoic acid (DHA) (22:6).In some cases, 60% to 75% Triglyceride oil be it is a kind of in glyceride form 18:1 lipid.In one embodiment, the triglyceride oil is less than 2%14:0th, 13% to 16%16:0th, 1% to 4%18:0th, 64% to 70%18:1st, 10% to 16%18:2nd, 0.5% to 2.5%18:3 and carbon chain lengths less than 2% be 20 or longer oil.

In some cases, the biomass is carbohydrate by dry weight between 25% and 40%.In some feelings Under condition, the carbohydrate ingredient of the biomass is dietary fiber and 2% to 8% by dry weight between 25% to 35% Between free sugar (including sucrose).In one embodiment, the monose of the dietary fiber component of the biomass, which is constituted, is 0.1% to 3% arabinose, 5% to 15% mannose, 15% to 35% galactolipin and 50% to 70% Portugal Grape sugar.In some cases, the biomass has the total carotinoid between 20 to 115 μ g/g, includes 20-70 μ g/g leaf Flavine.In one embodiment, the chlorophyll content of the biomass is less than 2ppm.In some cases, the biomass has 1-8mg/100g total tocopherols, include 2-6mg/100g alpha tocopherol.In some cases, the biomass has 0.05- 0.30mg/g total tocotrienols, includes 0.10-0.25mg/g α tocotrienols.

In some cases, microalgae of the biomass from heterotrophic growth.In some cases, the microalgae biomass is source In the microalgae of a kind of Chlorella.In one embodiment, the microalgae is Chlorella protothecoides (Chlorella Protothecoides a strain).In some cases, the microalgae biomass is derived from a single strain of microalgae.One In a little embodiments, the microalgae biomass is derived from the colour mutant for having pigment formation reduced compared with from its derivative strain A kind of algae.In one embodiment, the microalgae is to be deposited on October 13rd, 2009 in the case where preserving title PTA-10397 The Chlorella protothecoides 33-55 of American type culture collection.In one embodiment, the microalgae is in October, 2009 The Chlorella protothecoides 25-32 for American type culture collection being deposited in the case where preserving title PTA-10396 in 13rd.

At the 18th aspect, food ingredients group formed by egg products are combined with algae powder is passed through the invention provides one kind Compound, it is the homogenate of microalgae biomass, and the microalgae biomass mainly or entirely contains the dissolving cell in powder type, bag Include by dry weight at least 16% triglyceride oil, for being matched somebody with somebody by adding liquid and optional other comestible compositions Food article processed.In one embodiment, the food ingredients composition is a kind of pasta.

At the 19th aspect, the invention provides a kind of food ingredients combination including liquid egg products and algae slurry material Thing, wherein the algae powder are the homogenate of microalgae biomass, and the microalgae biomass mainly or entirely contains the dissolving in powder type Cell, including at least 16% triglyceride oil by dry weight.In some cases, the liquid egg products are liquid shell egg, liquid Body protein, liquid yolk or liquid egg substitute.In one embodiment, the food ingredients composition is used for scrambled egg product Prepare (in heating).

At the 20th aspect, dry egg is included the invention provides a kind of method for preparing food article, including by one kind The food ingredients and microalgae powder of product are combined and cooked with liquid and optional other comestible compositions, and the microalgae powder is The homogenate of microalgae biomass, mainly or entirely containing the dissolving cell in powder type, including by dry weight at least 16% Triglyceride oil.In some cases, the food article is powdery egg products or pancake or waffle powder.

The 20th on the one hand, the invention provides a kind of method for preparing food ingredients composition, including microalgae is provided The homogenate of biomass and liquid egg products, and make the homogenate dry together with liquid egg products to provide food ingredients combination Thing, the microalgae biomass mainly or entirely contains dissolving cell and by dry weight at least 16% triglyceride oil.In some feelings Under condition, this method further comprises making algal biomass micronizing provide the homogenate.In some cases, the food ingredients are combined Thing is the preparation for scrambled egg product (in heating).

At the 22nd aspect, the invention provides a kind of by egg products and slurry, the Yi Jizhi of microalgae powder or microalgae powder A kind of few other comestible compositions are combined and food ingredients composition formed by heating, and the wherein microalgae powder is microalgae life The homogenate of material, the microalgae biomass mainly or entirely contains the dissolving cell in powder type, including by dry weight at least 16% triglyceride oil.In one embodiment, the egg products are liquid egg products.In some cases, the liquid egg Product is liquid shell egg, liquid yolk, liquid protein or liquid egg substitute.In one embodiment, the egg products are dry Dry egg products.In some cases, the egg products of the drying are the albumen of dry shell egg, the yolk of drying or drying. In some embodiments, at least one others comestible composition includes a kind of edible liquid.In one embodiment, The food composition is scrambled egg.

At the 23rd aspect, the invention provides a kind of food ingredients composition including egg products and algae powder, algae powder It is the homogenate of microalgae biomass, the microalgae biomass mainly or entirely contains the dissolving cell in powder type, including by dry Triglyceride oil and by dry weight at least 40% protein of the restatement no more than 20%, for by add edible liquid with Optional other comestible compositions prepare food article.

The present invention further comprises uniqueness and novel microalgae strain, and the strain of these microalgaes is already subjected to non-transgenic method Mutation is enough to reduce the coloring of the biomass produced by these plants.The biomass produced from these plants can be used for manufacture to bakee food Thing, the food without seitan, beverage, high fat algae powder and other foods.Pigment such as carotenoid and chlorophyll are combining Enter food (such as mayonnaise, Yoghourt and white sauce, i.e., unconventional related color such as yellow, red, orange and green) When for consumer receive be probably undesirable.Some pigments (such as chlorophyll) can also produce undesirable taste profile. The scope for the food article with healthy lipodogramme that can be manufactured is expanded using hypopigmented microalgae biomass.Same tool The biomass of the invention containing high protein for the pigment formation being reduced is also incorporated into product (such as textured vegetable protein, nutrition bar And meal replacement beverage) in.The reduced microalgae of pigment formation also allows the biomass with reference to higher amount to enter some foods In Tetramune, it otherwise can use the microalgae biomass of high coloring and realize.It there is disclosed herein and produce the hypopigmented of novelty The method of microalgae.Strain provided by the present invention is also useful in the triglyceride oil of the extraction of the muted color of manufacture health.

In twenty-fourth aspect, the invention provides a kind of food composition, the microalgae comprising at least 0.1%w/w is biological Matter and one or more other comestible compositions, wherein microalgae biomass include by dry weight at least 16% triglycerides Oil, and be the reduced color change body of pigment formation there is provided the strain of the microalgae of the biomass compared with from its derivative strain.One In the case of a little there is provided the strain of the microalgae of the biomass have with the Chlorella protothecoides grown under comparable conditions compared with reduction Color.In one embodiment, microalgae strain is to be deposited in the U.S. in the case where preserving title PTA-10397 on October 13rd, 2009 The Chlorella protothecoides 33-55 of Type Tissue Collection.In one embodiment, microalgae strain is October 13 in 2009 Day is deposited in the Chlorella protothecoides 25-32 of American type culture collection in the case where preserving title PTA-10396.In some feelings Grow and process under the conditions of Good Manufacture Practice (GMP) there is provided the strain of the microalgae of the biomass under condition.

In some embodiments, the food composition is to be selected from the group, and the group is made up of the following：Salad seasoning Juice, egg products, baked goods, bread, food bar, pasta, sauce, soup drink (soup drink), beverage, freezing sweet tea Product, dough/pasta, butter substitute smear food.In some cases, one or more comestible compositions are to be selected from the group, The group is made up of the following：Cereal, water fruits and vegetables, protein, herbaceous plant or spices.In one embodiment, the food Compositions further comprise a kind of food compatibility preservative.

At the 25th aspect, the invention provides a kind of food composition, the microalgae comprising at least 0.1%w/w is biological Matter and one or more other comestible compositions, the wherein microalgae biomass comprising by dry weight at least 40% protein simultaneously And be from it is a kind of be color change body microalgae strain prepare, compared with from its derivative strain, the microalgae strain have reduced color Element is formed.In one embodiment, microalgae strain is to be deposited on October 13rd, 2009 in the case where preserving title PTA-10397 The Chlorella protothecoides 33-55 of American type culture collection.In one embodiment, microalgae strain is in 2009 10 The moon 13 was deposited in the Chlorella protothecoides 25-32 of American type culture collection in the case where preserving title PTA-10396.

At the 26th aspect, the invention provides a kind of method, the one kind for being suitable for food production this method provide Microalgae strain, including a kind of microalgae strain of (a) mutagenic treatment, (b) has when identification grows under the same conditions relative to prime strain The reduced mutagenesis bacterium colony of pigment formation；And (c) is providing by dry weight at least 25% triglycerides oil content and/or by thin The mutagenesis bacterium colony is cultivated under conditions of the protein content of born of the same parents' dry weight meter at least 40%.In some embodiments, this method is entered One step includes collecting cultivated cell, and by the microalgae biomass roller drying.In some cases, the microalgae of the drying Biomass includes the total carotinoid less than 5mcg/g.In one embodiment, the microalgae biomass of the drying is included and is less than 2mcg/g total carotinoid.In one embodiment, the microalgae biomass of the drying includes total class less than 1.1mcg/g Carrotene.

In some cases, this method is carried out with the microalgae of a kind of Chlorella.In an embodiment In, microalgae strain is Chlorella protothecoides (Chlorella protothecoides).In some cases, the mutagenic fungi is heterotrophism Culture.In some embodiments, the mutagenic fungi being capable of heterotrophic growth.In one embodiment, the microalgae plant is The Chlorella protothecoides of American type culture collection are deposited in the case where preserving title PTA-10397 on October 13rd, 2009 33-55.In one embodiment, microalgae strain is to be deposited on October 13rd, 2009 in the case where preserving title PTA-10396 The Chlorella protothecoides 25-32 of American type culture collection.

The 27th aspect, the invention provides prepare food article a kind of method, including by microalgae biomass with One or more others comestible composition combinations, wherein microalgae biomass includes by dry weight at least 16% triglycerides Oil and/or at least 40% protein by dry weight, and the microalgae biomass and the protoconch bead that grows under the same conditions The biomass of algae compares the coloring with reduction.In some cases, the food article is to be selected from the group, and the group is by following Items composition：It is salad-dressing, egg products, baked goods, bread, food bar, pasta, sauce, soup drink, beverage, cold Freeze sweets, dough/pasta, butter substitute or smear food.

In twenty-eighth aspect, the invention provides a kind of food ingredients composition, it includes the homogenate of microalgae biomass, Microalgae biomass mainly or entirely contains the dissolving cell in powder type, including at least 10% triglycerides by dry weight Oil, and it is the colour mutant for having pigment formation reduced compared with from its derivative strain to provide the microalgae strain of the biomass. In some cases, microalgae strain grows and processed under the conditions of Good Manufacture Practice (GMP).In some embodiments In, the microalgae biomass is derived from the algae of Heterotrophic culture.In some cases, the microalgae biomass includes existing by dry weight Oil between 45% and 70%.In one embodiment, the microalgae biomass includes by dry weight at least 40% protein.

In some cases, food composition further comprises antioxidant.In some cases, food ingredients composition Further comprise glidant.

In some embodiments, the food composition includes the biomass from microalgae strain, and microalgae strain is chlorella One microalgae species of category.In one embodiment, microalgae strain is Chlorella protothecoides (Chlorella protothecoides).In one embodiment, microalgae strain is to preserve title PTA- on October 13rd, 2009 The Chlorella protothecoides 33-55 of American type culture collection is deposited under 10397.In one embodiment, the microalgae Strain is that the protoconch that American type culture collection is deposited in the case where preserving title PTA-10396 on October 13rd, 2009 is small Ball algae 25-32.

The present invention further comprises including the finished product food composition of seitan reduction and without seitan microalgae, together with In the food ingredients containing microalgae of manufacture seitan reduction on a large scale and without seitan food.The present invention food and into Point, while seitan is reduced or eliminated, also with the replacement via primary single unsaturated algae oil by reduce or eliminate compared with Unsound oil ＆ fat and increased health benefits.These novel food compositions and the not gluten containing food being previously present Compare, also with more desirable organoleptic attribute and shelf-life.Food and composition disclosed here are free of bran with existing The food of matter compares, containing the seitan or no seitan being reduced, and contains high dietary fiber level, the cholesterol of reduction Or eliminate cholesterol and more healthy oil content.Further disclose reduction food hypersenstivity and disease symptomses (such as stomatitis abdomen Rush down) method to address the cumulative Sensitivity rate to the product containing seitan.Further disclose preparation and manufacture contains microalgae Food without seitan and for the method for the composition for preparing such food.

At the 29th aspect, the invention provides food article formed by a kind of combination by microalgae biomass, The microalgae biomass is including at least 16% triglyceride oil by dry weight and at least one other powder for being free of seitan or not Cereal products containing seitan.In some cases, should be free of the powder of seitan or the cereal products without seitan is included below at least One of：Three-coloured amaranth powder, kudzuvine root starch, buckwheat, ground rice, olecranon bean powder, corn flour, millet powder, dehydrated potato powder, farina, Kui Nu Sub- lamb's-quarters powder, sorghum flour, soy meal, bean powder, legume flour, cassava (cassava) powder, eragrosits abyssinica powder, globe artichoke Powder, almond powder, acorn flour, coconut powder, chest nut powder, corn flour and taro meal.

In some cases, the food article is formed with microalgae biomass, the microalgae biomass be in microalgae thin slice, Algae powder or a kind of form of microalgae powder, it is the homogenate of microalgae biomass or by the way that the powder is dispersed in into a kind of edible liquid Slurry formed by body, the microalgae biomass mainly or entirely contains the dissolving cell in powder type.In certain situation Under, the microalgae biomass is mainly dissolving cell.In one embodiment, the microalgae biomass is microalgae powder.In some feelings Under condition, the microalgae powder has the particle mean size between 1 and 100 μm.In one embodiment, the triglyceride oil is less than 2%14:0th, 13% to 16%16:0th, 1% to 4%18:0th, 64% to 70%18:1st, 10% to 16%18:2nd, 0.5% to 2.5%18:3 and carbon chain lengths less than 2% be 20 or longer oil.In some cases, the microalgae biomass has and pressed Carbohydrate of the dry weight meter between 25% and 40%.In some cases, the carbohydrate ingredient of the microalgae biomass It is between 25% to 35% dietary fiber by dry weight and 2% to 8% free sugar (including sucrose).In an embodiment party In case, the monose composition of the dietary fiber component of the biomass is 0.1% to 4% arabinose, 5% to 15% sweet dew Sugar, 15% to 35% galactolipin and 50% to 70% glucose.In some cases, the microalgae biomass has 20 Total carotinoid to 115 μ g/g, includes 20-70 μ g/g lutein.In one embodiment, the microalgae is biological The chlorophyll content of matter is less than 200ppm.In one embodiment, the chlorophyll content of the microalgae biomass is less than 2ppm. Under certain situation, the biomass has 1-8mg/g total tocopherols, includes 2-6mg/g alpha tocopherol.In some cases, The biomass has 0.05-0.30mg/g total tocotrienols, includes 0.10-0.25mg/g α tocotrienols.One In the case of a little, the triglyceride oil is by weight the docosahexaenoic acid (DHA) (22 less than 5%:6).

In some embodiments, the microalgae biomass is the form in microalgae powder and the powder does not have visible oil. In some cases, the microalgae biomass is the form in microalgae powder and further comprises glidant.In some cases, The microalgae biomass is the form in microalgae powder and the powder further comprises antioxidant.

In different embodiments, the microalgae biomass is derived from the single strain for being no more than one of microalgae.At some In the case of, the microalgae biomass is derived from the microalgae of a kind of Chlorella.In one embodiment, the microalgae is former Shell chlorella (Chlorella protothecoides).In some cases, the microalgae biomass is derived from color change A kind of microalgae of strain, there is this microalgae reduced pigment to be formed compared with from its derivative strain.In some embodiments, should Microalgae biomass is derived from the microalgae of Heterotrophic culture.In some embodiments, microalgae biomass is derived from good production rule The algae cultivated and processed under the conditions of model (GMP).

In some cases, the food article is a kind of baked goods, bread, cereal preparation, crispbread or Italy Wheaten food.In some embodiments, the baked goods are to be selected from the group, and the group is made up of the following：Brownie, cake, And pie product and cookies.In one embodiment, the food article is free from seitan.In some cases, will Food compatibility preservative is added in the microalgae biomass.In some cases, the food article is free of except belonging to microalgae life Oily or fat beyond the algae oil of material.In some cases, the food article is free of yolk.In some embodiments, should Microalgae biomass has about 0.5% to 1.2%w/w algae phosphatide.In some cases, these phosphatide include phosphatidyl courage The combination of alkali, phosphatidyl-ethanolamine and phosphatidylinositols.In one embodiment, the food article is a kind of product of life. In one embodiment, the food article is a kind of ripe product.

At the 30th aspect, the invention provides a kind of compound powder without seitan, it is including a kind of microalgae powder and extremely A kind of few others in addition to microalgae powder are free of the powder of seitan, and the wherein microalgae powder includes the homogenate of microalgae biomass, and this is micro- Algal biomass mainly or entirely contains the dissolving cell in powder type, and contains by dry weight at least 16% glycerine three Ester oil.In some cases, at least one other powder without seitan are to be selected from the group, and the group is by the following group Into：Three-coloured amaranth powder, kudzuvine root starch, buckwheat, ground rice, olecranon bean powder, cornflakes, cornflour, millet powder, dehydrated potato powder, potato are formed sediment Powder, quinoa powder, sorghum flour, soy meal, bean powder, legume flour, cassava (cassava) powder, eragrosits abyssinica powder, Globe artichoke powder, almond powder, acorn flour, coconut powder, chest nut powder, corn flour and taro meal.In some embodiments, this is micro- The particle mean size of the particle of biomass in algae powder is between 1 and 100 μm.In some cases, the microalgae powder has by weight Gauge 10% or less or 5% or less moisture.In some cases, the microalgae biomass has and existed by dry weight Triglyceride oil between 45% and 70%.In one embodiment, 60% to 75% oil is a kind of in glyceride shape The 18 of formula:1 lipid.In one embodiment, the triglyceride oil is less than 2%14:0th, 13% to 16%16:0th, 1% to 4%18:0th, 64% to 70%18:1st, 10% to 16%18:2nd, 0.5% to 2.5%18:3 and the carbon chain lengths less than 2% For 20 or longer oil.

In some cases, should without seitan compound powder include microalgae biomass, it be by dry weight 25% with Carbohydrate between 40%.In some embodiments, the carbohydrate ingredient of the microalgae biomass is by dry weight The free sugar (including sucrose) between dietary fiber and 2% to 8% between meter 25% to 35%.In an embodiment In, the monose of the dietary fiber component of the biomass constitute be 0.1% to 4% arabinose, 5% to 15% mannose, 15% to 35% galactolipin and 50% to 70% glucose.In some cases, the microalgae biomass have 20 to Total carotinoid between 115 μ g/g, includes 20-70 μ g/g lutein.In one embodiment, the microalgae biomass Chlorophyll content be less than 200ppm.In some cases, the chlorophyll content of the microalgae biomass is less than 2ppm.In some feelings Under condition, the microalgae biomass has 1-8mg/100g total tocopherols, includes 2-6mg100g alpha tocopherol.In certain situation Under, the biomass has 0.05-0.30mg/g total tocotrienols, includes 0.10-0.25mg/g α tocotrienols.

In some embodiments, the microalgae powder does not have visible oil.In some cases, one should be entered without the powder of seitan Step includes glidant.In some cases, antioxidant should be further comprised without the powder of seitan.

In some cases, the microalgae biomass is derived from the single strain for being no more than one of microalgae.In some cases, The microalgae biomass is derived from the algae of a kind of Chlorella.In one embodiment, the algae is Chlorella protothecoides (Chlorella protothecoides).In some cases, the microalgae biomass is derived from compared with from its derivative strain A kind of algae with the reduced colour mutant of pigment formation.In some embodiments, the microalgae biomass is derived from heterotrophism The algae of culture.In some cases, microalgae biomass is derived from cultivating and processing under the conditions of Good Manufacture Practice (GMP) Algae.

The 30th on the one hand, the invention provides the method that mitigation seitan does not tolerate symptom, including (a) is with seitan Contain bran with the same type of food article substitution produced by combining microalgae biomass in the diet for the subject not tolerated The food article of matter, the microalgae biomass includes at least 16% triglyceride oil on dry basis and at least one others not Food ingredients containing seitan, the wherein same type of food article are free from seitan；And do not tolerated to seitan (b) Subject the same type of food article is provided, thus mitigate the disease that at least one seitan in the subject is not tolerated Shape.

At the 32nd aspect, the method for the food article without seitan is manufactured the invention provides a kind of, including pass through Microalgae biomass is set to be combined with least one other edible component without seitan to manufacture the food article, microalgae life Material includes by dry weight at least 16% triglyceride oil.In some cases, the microalgae biomass has and existed by dry weight Oil between 45% and 70%.In some cases, 60% to 75% triglyceride oil is a kind of in glyceride form 18:1 lipid.In one embodiment, the triglyceride oil is less than 2%14:0th, 13% to 16%16:0th, 1% to 4% 18:0th, 64% to 70%18:1st, 10% to 16%18:2nd, 0.5% to 2.5%18:3 and carbon chain lengths less than 2% be 20 Or longer oil.

In some cases, the microalgae biomass is derived from the single strain for being no more than one of microalgae.In some embodiment party In case, the microalgae biomass is derived from the algae of a kind of Chlorella.In one embodiment, the algae is that protoconch is small Ball algae (Chlorella protothecoides).In some cases, the microalgae biomass is derived from the algae of Heterotrophic culture. In some cases, microalgae biomass is derived from the algae cultivated and processed under the conditions of Good Manufacture Practice (GMP).

The present invention further comprises the method by providing the food based on microalgae and inducing satiety.In some embodiment party In case, microalgae biomass contains high-caliber dietary fiber and/or digestible thick protein and/or low saturation triglycerides Oil.The method that homogenizes of release free oil and fiber is disclosed, thus the sensation being satiated with food for strengthening in the mankind reduces heat Intake.The offer of this material has more benefits for the mankind：The composition based on microalgae of beneficial health of heart is provided with, together Shi Shixian is enough to reduce the level of being satiated with food of more energy intakes.

At the 33rd aspect, the invention provides a kind of method of the inducing satiety in the mankind, including one kind is given Food article including microalgae biomass, the microalgae biomass is combined with one or more other comestible compositions, wherein should Microalgae biomass includes at least 16% triglyceride oil and by dry weight at least 10% total dietary fiber by dry weight. Under certain situation, the microalgae biomass includes oil by dry weight between 45% and 70%.In some embodiments, 60% To 75% triglyceride oil be it is a kind of in glyceride form 18:1 lipid.In one embodiment, the triglycerides Oil is less than 2%14:0th, 13% to 16%16:0th, 1% to 4%18:0th, 64% to 70%18:1st, 10% to 16%18:2、 0.5% to 2.5%18:3 and carbon chain lengths less than 2% be 20 or longer oil.

In some cases, the microalgae biomass has carbohydrate by dry weight between 25% to 45%. Under certain situation, dietary fiber between by dry weight 25% to 35% of the carbohydrate ingredient of the microalgae biomass and Free sugar (including sucrose) between 2% to 8%.In one embodiment, the monose of the dietary fiber component of the biomass Composition be 0.1% to 4% arabinose, 5% to 15% mannose, 15% to 35% galactolipin and 50% to 70% glucose.In some cases, the microalgae biomass include about 20% Soluble Fiber and about 10% not Soluble fiber.In some cases, in the microalgae biomass dietary fiber and the ratio of triglyceride oil is about 3:5. In some embodiments, the microalgae biomass has 20-115 μ g/g total carotinoid, includes 20-70 μ g/g leaf Huang Element.In one embodiment, the chlorophyll content of the microalgae biomass is less than 2ppm.In some cases, the microalgae is biological Matter has 1-8mg/100g total tocopherols, includes 2-6mg/100g alpha tocopherol.In one embodiment, the microalgae Biomass has 0.05-0.3mg/g total tocotrienols, includes 0.10-0.25mg/g α tocotrienols.

In some embodiments, one or more other edible components are to be selected from the group, and the group is by the following Composition：Cereal, water fruits and vegetables, protein, herbaceous plant and spices.In some cases, food article is to be selected from the group, The group is made up of the following：Egg products, food bar, baked goods, bread, pasta, soup, beverage and dessert. In one embodiment, the food article is a kind of nutritious drink for being suitable as dietary substitutes.In some embodiments, The microalgae biomass does not have visible oil.

In some cases, the microalgae biomass is processed to microalgae powder, and it is a kind of mainly or entirely contains in powder The homogenate of the dissolving cell of last form.In one embodiment, the powder further comprises glidant.In an embodiment In, the water content of the powder is by weight 10% or lower.In some cases, the average grain of the microalgae biomass in the powder Degree is between 1 and 100 μm.In one embodiment, the powder further comprises antioxidant.

In different embodiments, used microalgae biomass is derived from not surpassing for microalgae in the method for the invention Cross the single strain of one.In some cases, the microalgae biomass is derived from the microalgae of a kind of Chlorella.In a reality Apply in scheme, the microalgae is Chlorella protothecoides (Chlorella protothecoides).In some cases, the microalgae is biological Matter is derived from a kind of algae for the colour mutant for having pigment formation reduced compared with from its derivative strain.In some cases, The microalgae biomass is derived from the algae of Heterotrophic culture.In some embodiments, microalgae biomass is derived from well producing The algae cultivated and processed under the conditions of specification (GMP).

In some cases, food article includes at least 0.5%w/w microalgae biomass.In one embodiment, should Microalgae biomass includes at least 40% protein and the triglyceride oil no more than 20% by dry weight.In an embodiment In, the ratio of dietary fiber and protein in the microalgae biomass is about 3:10.In some cases, the microalgae is biological Matter includes about 10% Soluble Fiber and about 4% insoluble fibre by dry weight.In some cases, the microalgae is given birth to Material has the chlorophyll no more than 200ppm.In one embodiment, the protein source be at least 40% digest it is thick Albumen.In one embodiment, the microalgae biomass includes 1-3g/100 total sterol.

At the 34th aspect, the invention provides a kind of method of inducing satiety, it is included in the diet of subject One or more conventional food articles are substituted containing microalgae food article with same type of one or more, wherein this is same Type containing microalgae food article contains microalgae biomass, and the microalgae biomass includes by dry weight at least 16% glycerine three Ester oil and by dry weight at least 10% total dietary fiber, wherein the calorie consumed by the subject is with substituting the identical of diet Or it is lower, and the subject has increased satietion.In some cases, the microalgae biomass has 45% to 70% Triglyceride oil.In one embodiment, in the microalgae biomass dietary fiber and the ratio of triglyceride oil is big About 3:5.In one embodiment, the microalgae biomass further comprises by dry weight at least 40% protein.At some In embodiment, the ratio of dietary fiber and protein in the microalgae biomass is about 3:10.In some cases, contain There is microalgae food article to include at least 0.5%w/w microalgae biomass.

In some cases, conventional food article is to be selected from the group, and the group is made up of the following：Egg products, food Rod, baked goods, bread, pasta, soup, beverage and dessert.In one embodiment, the beverage is a kind of is adapted to It is used as the nutritious drink of dietary substitutes.In some cases, containing microalgae food article compared with conventional food article When, oil, fat or eggs with identical or reduction.

At the 35th aspect, the invention provides a kind of method of the inducing satiety in subject, including to tested Person gives a kind of microalgae food article, and wherein the microalgae food article is comparable to conventional food article, except with by dry weight The microalgae biomass of at least 10% total dietary fiber is replaced normal including at least 16% triglyceride oil and by dry weight Outside some or all of oil, fat or egg in the food article of rule.

The present invention further comprises protein and the high microalgae biomass of fiber content, and wherein the biomass is by different Hair care ferment and manufacture.Provided herein these materials for manufacture meat analogue product and meat reinforcing agent benefit from addition together with other can The food article of digesting protein and dietary fiber is useful.The architectural characteristic of food by using such Material reinforcement (including texture and water-retaining property).The present invention high protein and fiber foodstuff can from it is edible and be unsuitable for food Heterotrophic fermentation raw material (including cornstarch, sugarcane, glycerine and depolymerizing cellulose) is manufactured.

At the 36th aspect, the invention provides a kind of microalgae powder, it is mainly or entirely to contain to be in powder type Dissolving cell microalgae biomass homogenate, the wherein algal biomass includes at least 40% protein and pressing by dry weight Dry weight meter is less than 20% triglyceride oil, also, wherein the algal biomass is derived under the conditions of Good Manufacture Practice (GMP) Heterotrophic culture and processing algae.In some cases, the particle mean size of particle is less than 100 μm.In some cases, powder The particle mean size of particle in end is 1-15 μm.In some embodiments, the powder is by the way that microalgae biomass is micronized Form a kind of emulsion and make formed by emulsion drying.In one embodiment, the microalgae powder has by dry weight 10% or lower water content.In some cases, the algal biomass includes by dry weight at least 20% carbohydrate. Under certain situation, the algal biomass includes by dry weight at least 10% dietary fiber.In one embodiment, the protein Be at least 40% digestible crude protein.

In some embodiments, microalgae biomass is derived from the algae of Heterotrophic culture.In some cases, the biomass It is derived from the algae of a kind of Chlorella.In one embodiment, the algae is Chlorella protothecoides (Chlorella protothecoides).In some cases, the algal biomass is derived from the single strain for being no more than one of microalgae.At some In embodiment, the algal biomass is without the Algae toxins that can be detected.In one embodiment, the chlorophyll content of the biomass Less than 200ppm.In some cases, the biomass includes 1-3g/100 total sterol.In some cases, the biomass bag Total tocopherols containing 0.15-0.8mg/100g, include 0.18-0.35mg/100g alpha tocopherol.In some embodiments, The biomass is derived from a kind of algae for the colour mutant for having pigment formation reduced compared with from its derivative strain.

In some cases, the microalgae powder further comprises food compatibility preservative.In one embodiment, the food Thing compatibility preservative is a kind of antioxidant.

At the 37th aspect, the invention provides a kind of food ingredients, it includes microalgae powder discussed above with being adapted to In the other protein product combinations of at least one of human consumption, the wherein food ingredients include by dry weight at least 50% egg White matter.In some embodiments, at least one other protein products are derived from vegetarian diet source.In some cases, the element It is to be selected from the group to eat source, and the group is made up of the following：Soybean, pea, bean or pea, milk, whey, rice and wheat.

In some cases, the microalgae biomass of the food ingredients is derived from the algae of a kind of Chlorella.One In individual embodiment, the algae is Chlorella protothecoides (Chlorella protothecoides).In some cases, the microalgae Biomass is derived from the algae of Heterotrophic culture.In some cases, the microalgae biomass is derived from compared with from its derivative strain A kind of algae with the reduced colour mutant of pigment formation.

At the 38th aspect, the invention provides by the way that microalgae powder discussed above and at least one others is edible With into a kind of food composition formed by subassembly.In some cases, the food composition is the meat generation of vegetarian a kind of Articles for use, protein rod or nutritious drink.

The 39th aspect, the invention provides by by including at least 40% protein by dry weight and less than by The microalgae biomass of the triglyceride oil of dry weight meter 20% combine with least one other comestible compositions formed by one kind It is that food composition, the wherein algal biomass are derived under the conditions of Good Manufacture Practice (GMP) Heterotrophic culture and processing Algae.In some cases, the microalgae biomass is to be in microalgae thin slice, algae powder, the form of algae powder, and it is microalgae biomass Homogenate or by slurry formed by the algae powder is dispersed in a kind of edible liquid, the microalgae biomass is mainly or entirely Contain the dissolving cell in powder type.In some cases, the microalgae biomass is algae powder or slurry.In some embodiment party In case, at least one others comestible composition is meat products.In some cases, the food composition is raw product. In some cases, the food composition is ripe product.

At the 40th aspect, the invention provides a kind of method for the meat analogue product for manufacturing vegetarian, this method includes：Will The microalgae biomass of triglyceride oil including at least 40% protein by dry weight and less than by dry weight 20% with least A kind of other vegetarian's protein source combinations, also, wherein the microalgae biomass is derived from Good Manufacture Practice (GMP) condition Lower Heterotrophic culture and processing microalgae.

The 40th on the one hand, the invention provides it is a kind of manufacture crush meat products method, including by meat products with The microalgae biomass combination of triglyceride oil including at least 40% protein by dry weight and less than by dry weight 20%, and And wherein the algal biomass is derived under the conditions of Good Manufacture Practice (GMP) Heterotrophic culture and processing microalgae.

At the 42nd aspect, the invention provides one kind by by microalgae biomass and at least one comestible composition group A kind of food composition formed by conjunction, the microalgae biomass includes by dry weight at least 13% total dietary fiber.At some In the case of, the algal biomass includes total dietary fiber by dry weight between 13% to 35%.In some cases, the microalgae Biomass is included in the Soluble Fiber between 10% to 25%.In some cases, microalgae biomass is included in 4% to 10% Between insoluble fibre.

At the 43rd aspect, the invention provides a kind of method for manufacturing algae protein concentrate, including (a) will be by dry weight Meter includes the microalgae biomass degreasing of at least 40% protein, and (b) removes soluble sugar from the degreasing microalgae biomass, by This produces a kind of algae protein concentrate.

At the 44th aspect, the invention provides a kind of algae protein concentrate manufactured by the following method, including (a) The microalgae biomass degreasing of at least 40% protein by dry weight will be included, and (b) can from degreasing microalgae biomass removal Dissolubility sugar, thus produces a kind of algae protein concentrate.

At the 45th aspect, the invention provides a kind of algae Protein Separation thing, wherein minimum protein content is by dry Restatement 90%, and be from including at least the microalgae biomass of 40% protein is produced by dry weight.

The present invention further comprises the triglyceride oil for human consumption of novelty.Traditionally, for example low mustard of Agricultural Materials Sauerkraut seed, soybean and olive are always the source of edible oil, and these materials are that (wherein these crops can plant by geography Training) limitation.The oil of the present invention can be (including cornstarch, sweet from edible and inedible heterotrophic fermentation raw material Sugarcane, glycerine and depolymerizing cellulose) and manufacture, these raw materials are the purpose growth either geographic areas from non-constant width The byproduct of multifarious existing agricultural procedure.Food oil disclosed here is low saturation, high monounsaturated, and can be led to Cross and manufactured using the strain of hypopigmented microalgae in hypopigmented form.Food oil disclosed here can be by using many Plant different types of oil-producing microalgae and manufacture.

At the 46th aspect, the invention provides a kind of microalgae triglyceride oil for the purifying for being suitable for human consumption, It includes at least 50% oleic oil and DHA less than 5%, and wherein microalgae oil is without the microalgae toxin that can be detected and is Prepared under good manufacturing condition.In some cases, culinary applications can be suitable for by triglyceride oil being packaged in Bottle or spray in.In some cases, the oil is packed with the volume of the oil product more than 50mL.In a reality Apply in scheme, the oil is less than 2%14:0th, 13% to 16%16:0th, 1% to 4%18:0th, 64% to 70%18:1st, 10% to 16%18:2nd, 0.5% to 2.5%18:3 and carbon chain lengths less than 2% be 20 or longer oil.

In some cases, microalgae oil is purified from the single strain no more than one of microalgae.In certain situation Under, the microalgae is a kind from Chlorella.In one embodiment, the microalgae is Chlorella protothecoides (Chlorella protothecoides)。

In some embodiments, the microalgae triglyceride oil further comprises a kind of antioxidant of addition.At some In the case of, the oil has the total carotinoid between 40-230 μ g/g, includes 40-70 μ g/g lutein.In an embodiment party In case, the oil has the chlorophyll less than 2ppm.In some cases, the oil has 2-16mg/100g total tocopherols, bag Include 4-12mg/100g alpha tocopherol.In some cases, the oil has 0.10 to total tocotrienols between 0.6mg/g Class, includes 0.2-0.5mg/g α tocotrienols.

At the 47th aspect, food, including microalgae glycerine as claimed in claim 1 are smeared the invention provides one kind Three ester oils and a kind of liquid, the wherein oil and the liquid are formed a kind of emulsion of stabilization.In one embodiment, the smearing Food further comprises emulsifying agent.In some cases, the smearing food can be smeared at ambient temperature.In certain situation Under, the smearing food can be smeared at 5 DEG C -10 DEG C.

At the 48th aspect, the invention provides a kind of margarine, by making the life under the conditions of Good Manufacture Practice The microalgae triglyceride oil of the purifying of production is subjected to a kind of chemical reaction or enzymatic reaction, thus produces the margarine.At some In the case of, the chemical reaction is hydrogenization.In some cases, the chemical reaction or enzymatic reaction be with from microalgae glycerine The glyceride of the different lipodogrammes of three ester oils carries out ester exchange.In some cases, the different fat from microalgae triglyceride oil These mass spectrographic glyceride come from the one or more oil being selected from the group, and the group is made up of the following：Soybean, rape Seed, canola (canola, Canola), palm, palm kernel, coconut, corn, olive, sunflower, cottonseed, calyx are away from flower, flower Life, False flax, mustard seeds, cashew nut, oat, lupin, mestha, pot marigold, hemp, coffee, linseed, fibert, the root of Beijing euphorbia, pumpkin Seed, coriander, camellia, sesame, safflower, paddy rice, tung tree, cocoa, copra, opium poppy, castor bean, hickory nut, Xi Mengde Wood, manioca, Queensland nut, Bertholletia excelsa and avocado.

At the 49th aspect, the invention provides a kind of triglyceride oil for the purifying for being suitable for human consumption, wherein The oil is to be purified from microalgae and be mainly liquid at 4 DEG C, and wherein the oil is without the microalgae toxin that can be detected and is Prepared under good manufacturing condition.

At the 50th aspect, the invention provides the microalgae triglyceride oil of purifying, wherein the oil is no can detect level Phosphatide and chlorophyll having less than 2ppm.In some cases, the oil further comprises at least one：(a) it is big About 12-13ppm natural TBHQ (TBHQ)；(b) 1.34% free fatty；(c) it is less than 0.1% karr Take not humidity；(d) it is less than 0.1% monoglyceride；(e) it is less than 3% diglyceride；(f) about 6mg/100g total fertility Phenol, includes about 5.58mg/100g alpha tocopherol；And (g) about 0.24mg/g total tocotrienols.

The 50th on the one hand, it is suitable for the side of the microalgae triglyceride oil of human consumption the invention provides a kind of manufacture Method, including (a) is from the microalgae biomass extract oil for the triglyceride oil for including at least 25% by dry weight, and (b) carries this The oil taken is subjected to one or more following steps：Removing free fatty, bleaching, and deodorization, the wherein microalgae biomass is Grow and process under the conditions of Good Manufacture Practice (GMP), and wherein the triglyceride oil is less than 2%14:0th, 13% To 16%16:0th, 1% to 4%18:0th, 64% to 70%18:1st, 10% to 16%18:2nd, 0.5% to 2.5%18:3 and few In 2% carbon chain lengths be 20 or longer oil.In some cases, it is no more than 180 °F from microalgae biomass extract oil At a temperature of carry out.In some embodiments, this method is carried out under good working condition (GMP).

At the 52nd aspect, the invention provides a kind of swelling agent for being suitable for human consumption, it is included in good life The degreasing microalgae biomass without the microalgae toxin that can be detected prepared under the conditions of production.In some cases, the swelling agent is tied Close and enter in a kind of baked goods.In one embodiment, the swelling agent is incorporated into a kind of beverage.

At the 53rd aspect, the invention provides by the way that swelling agent discussed above and at least one others is edible With into a kind of food article formed by subassembly.

At the 54th aspect, the invention provides a kind of degreasing microalgae biomass without the microalgae toxin that can be detected, Wherein the microalgae biomass is cultivated and processed under good working condition.

At the 55th aspect, the invention provides a kind of animal food product, it by by degreasing microalgae biomass with One or more others comestible compositions are combined and formed, and wherein the degreasing microalgae biomass constitutes the animal food product By dry weight at least the 0.1% of all the components.In some cases, one or more other comestible compositions include one kind Cereal.In one embodiment, the animal food is formulated for farm-animals.

These and other aspects of the invention and embodiment description are in accompanying drawing (Brief Description Of Drawings immediately below) Neutralize in following detailed description of the invention, and illustrated in the following example.Above and through this Shen Any or all feature please discussed can be combined in the different embodiments of the present invention.

Brief description of the drawings

Fig. 1 shows the lipodogramme of the microalgae strain of selection, is expressed as total lipid content percentage.Corresponding to each strain number Kind/strain is shown in the table 1 of example 1.

Fig. 2 shows the aminogram of the Chlorella protothecoides biomass compared to the aminogram of whole-egg protein matter.

Fig. 3, which is shown, to be had and the egg pulp without algae powder keeps the sensory evaluation scores of 60 minutes in steam table.Every 10 Minute is estimated to the outward appearance, texture and mouthfeel of egg.

Fig. 4 shows the algae powder (about 50% lipid by dry weight) under an optical microscope in aqueous dispersion.This A little arrows point to the single algae powder particles of particle mean sizes, and larger arrow point to it is agglomerating after dispersion is formed or into The algae powder particles of cluster together.

Fig. 5 A to C show and softly mixed at (5A)；(5B) homogenizes under 300 bar pressures；And (5C) is in 1000 bars The size distribution of algae powder particles is resuspended in water after being homogenized under pressure at once.

Fig. 6 shows the food article containing algae powder, full adipose versus, low fat control and one kind of fat-free control The result of sense organ team for evaluation.

Detailed description of the invention

Reader, is divided into some sections and trifle by detailed description of the invention for convenience.Section I provides as used herein The definition of different terms.Section II, in A-E parts, describes the method for preparing microalgae biomass, including：Suitable biology Body (A), produces the method (B) for lacking the microalgae strain that pigment formation or pigment formation are substantially reduced, condition of culture (C), concentration bar Part (D), and biomass produced by the invention chemical composition (E).Section iii, in A-D parts, describing will be micro- Algae thin slice (A), algae powder (B), the algae powder (C) of algal biomass processing cost invention；With the method for algae oil (D).Iv section is described The various foods of the present invention and the method for combining microalgae biomass with other food ingredients.

All processes described herein can be carried out according to GMP or equal regulations.In the U.S., on manufacture, packaging or guarantor The GMP regulations for depositing the food of people are incorporated into code 21C.F.R.110.For all purposes hereby by these regulations cited therein And supplemental provisions are combined by quoting with its full text.The GMP regulations in the U.S. and the equal regulation of other jurisdictions are fitted For determine food whether be it is adulterated (food be not suitable for food under conditions of produce), or food preparation, bag Whether dress or preservation are carried out under the conditions of antihygienic, or even it may be contaminated, or may be with other side Formula insalubrity.GMP provides to include observing following management rule：Disease control；Cleaning and staff training；Building Maintenance and hygienic operation with facility；The enough sanitary installations of offer and accommodation；Design, construction, the dimension of equipment and apparatus Shield and cleaning；Appropriate Quality Control Procedure is provided, to ensure reception, the inspection according to appropriate sanitary principle in food article All reasonable precautions are taken when testing, transport, isolate, prepare, produce, pack and storing, so as to prevent from contacting any dirt Dye source；And finished product food is stored and transported under certain conditions, described condition can protect food to prevent by physics, change Learn pollution or bad microorganism pollution and prevent the rotten deterioration of food and container.

I. define

Unless hereinafter stated otherwise, all scientific and technical terminologies as used herein all have the skill of the technical field of the invention The implication that art personnel are generally understood that.The general definition of many terms used herein is found in below with reference in document： Singleton etc., Dictionary of Microbiology and Molecular Biology (second edition, 1994)； The Cambridge Dictionary of Science and Technology (Walker is edited, 1988)；The Glossary of Genetics, the 5th edition, R.Rieger etc. (editor), Springer Verlag (1991)；And Hale And Marham, The Harper Collins Dictionary of Biology (1991).

" area percentage " refers to the area of the peak value observed using FAME GC/FID detection methods, wherein at this Each aliphatic acid is converted to fatty acid methyl ester (FAME) before testing in sample.For example, with any other aliphatic acid such as C14:1 compares, for a kind of without undersaturated 14 carbon atom (C14:0) it, observed an independent peak.For each class FAME peak areas are directly proportional to its percentage in the mixture and are based on all peaks being present in the sample Sum total and calculate (that is, [gross area at region/all measurements peak under specific peak] x 100).When refer to the present invention During the lipid profile of oil and cell, " at least 4%C8-C14 " refers in the cell or in the glyceride composition of the extraction At least 4% total aliphatic acid, which has, includes 8, the chain length of 10,12 or 14 carbon atoms.

" sterile " refers to the culture for the organism not polluted by other live organisms.

" baked goods " are referred to being commonly found in being prepared in bakery by using baking box and usually contain leavening agent Food.Baked goods include but is not limited to Brownie, cookies, pie, cake and cake.

" bioreactor " and " fermentation tank " represents locking device or the portion for cell is cultivated in suspension Divide locking device, such as fermentation tank or round.

" bread " represents the food containing powder, liquid and common leavening agent.Bread typically by drying in an oven Roasting is standby, although other cooking methods are also acceptable.The property of leavening agent can be chemical property or organic/biological Property.Typically, organic leavening agent is yeast.In the situation of the leavening agent (such as baking powder and/or sodium bicarbonate) using chemical property Under, these food articles are referred to as " quick bread ".Crispbread and other wafer dry sample products are free from leavening agent The example of bread.

" cellulosic material " represents the product of cellulose digestion, particularly glucose and xylose.Cellulose digestion is typically Other compound, such as disaccharides, oligosaccharide, lignin, furfural and other compounds can be produced.The source of cellulosic material includes (for example) but it is not limited to bagasse, beet pulp, maize straw, wood chip, sawdust and switchgrass.

" co-cultivation " and its version (such as " cultivate " and " common fermentation ") expression under culture conditions altogether, two or more The cell of type is present in same bioreactor.For the purposes of the present invention, both or more types is thin Born of the same parents are typically microorganisms, are typically all microalgaes, but can include a kind of non-microalgae cell type in some cases. Under certain situation, the condition of culture for being suitable for co-culturing includes two or more cell types can be promoted to grow and/or breed Condition, in other cases, including only promote one kind or hypotype in two or more cells growth and/or propagation and The condition of cell growth is maintained to remaining cell.

" co-factor " represent a kind of enzyme realize its enzymatic activity needed for substrate beyond molecule.

" conventional food product " is represented to be intended to for for example by the composition of human consumption, its do not have microalgae biomass or its His algae component, and containing composition generally relevant with food article, particularly vegetable oil, animal tallow and/or eggs, together with Other comestible compositions.Conventional food article includes the food article sold in shop and restaurant and the food made in family Tetramune.Conventional food article is typically recipe making routinely, and conventional recipe provides to include the oil from non-algae source Or fat and/or eggs are together with other comestible compositions.

" ripe product " represents (for example) to heat the food of a period of time in an oven.

" creaminess salad-dressing " be expressed as with high viscosity ease up slow discharge rate stabilising dispersions salad adjust Taste juice.In general, creaminess salad-dressing is opaque.

" cultivation ", " culture " and " fermentation " and their version represent using condition of culture promote intentionally it is a kind of or The growth and/or breeding of various kinds of cell (typically microalgae).The condition drafted excludes the growth of microorganism in its natural state And/or breeding (intervening without direct people).

" cell dissolving " represents dissolving of the cell in hypotonic environment.Cell dissolving is due to water penetration or is moved to cell Inside reach overly hydrated's state so that cell can not bear internal water osmotic pressure and therefore rupture.

" dietary fiber " refers to seeing the non-starch class in plant and other biological body (including microalgae) containing cell membrane Carbohydrate.Dietary fiber can be soluble (being dissolved in water) or insoluble (can not be dissolved in water).It is soluble and insoluble Property fiber constitute total dietary fiber.

" degreasing meals " represent to have undergone the microalgae biomass of oil extract processing, therefore relative to the biology before oil extract Contain less oil for matter.Cell in degreasing meals is mainly lysed.Degreasing meals include using solvent (oneself Alkane) algal biomass that is extracted.

" digestible crude protein " is that trip can be converted to using part or after with stomach enzymic digestion in protein From the part of nitrogen (amino acid).Using gastric enzyme (such as pepsin) and sample is digested, the free amino acid after measurement digestion, by This completes the in-vitro measurements of digestible crude protein.Measure feed/food sample in protein content and by sample-feed animal, The amount for the nitrogen collected in animal wastes is measured, the in-vivo measurement of digestible crude protein is thus completed.

" dry weight " and " dry cell weight " represents the weight determined under conditions of relative water shortage.For example, it is biological to mention microalgae Specific components of the matter comprising prescribed percentage by dry weight represent it is to be based on essentially all of water in biomass to be removed it The percentage that weight afterwards is calculated.

" comestible composition " represents edible any material or composition." comestible composition " includes but is not limited to paddy Thing, water fruits and vegetables, protein, herbaceous plant, spices, carbohydrate and fat.

" exogenous offer " represents to be supplied to the molecule of cell (including being supplied to the cell culture medium in culture).

" fat " expression is typically the lipid or lipid mixture of solid under usual indoor temperature and pressure." fat " Including but not limited to lard and butter.

" fiber " represents the non-starch carbohydrate of many sugar forms.Fiber can be water miscible or water-insoluble. Many microalgaes produce both soluble and insoluble fibres, and they are typically found in cell membrane.

" food finished product " and " finished product food ingredients " represent prepare packaging, using or consumption food composition.For example, " food finished product " can be curing, or constitute " food finished product " composition may be mixed with each other or otherwise that This is combined." finished product food ingredients " typically are applied in combination to form food article with other compositions.

" fixed carbon source " represents the carbon-containing molecules being present in solid or liquid form under environment temperature and pressure, typically It is organic molecule.

" food ", " food ingredients composition ", " food article " and " food material " represents to be intended to be or expected for mankind's work Any combinations thing absorbed for nutrition and/or calorie source.Food ingredients composition it is main by carbohydrate, fat, water and/or Protein is constituted, and constitutes essentially all of personal every daily caloric intake.The minimum that " food ingredients composition " has Weight can for typical tablet or capsules weight (weight of typical tablet/capsule is straight less than or equal to 100mg at least ten times To 1500mg)." food ingredients composition " is not encapsulation or tablet form.

" glyceride spectrum " represents the different carbon chain lengths and saturated level of the glyceride in specific biomass or oil sample sheet Distribution.For example, sample can have such glyceride spectrum, wherein about the 60% of glyceride is C18:1,20% is C18: 0,15% is C16:0, and 5% be C14:0.(such as " C when carbon length is generally mentioned:18 "), this formulation can include Arbitrary saturation capacity；For example, (being C containing 20% (by weight/quality meter) lipid:18) microalgae biomass can comprising equal or Different amounts of C18:0、C18:1、C18:2 etc., their summation constitutes the 20% of biomass.Mention the percentage of certain saturation type Than (the 18 of such as at least 50%:1 glyceride form it is single unsaturated) aliphatic lateral chain that refers to glyceride is the 18 of at least 50%: 1, but do not necessarily mean that at least 50% triglycerides is olein (three 18:1 chain is connected to single glycerine master Chain)；This spectrum can include having 18:1 with the glyceride of the mixtures of other side chains, condition is at least the 50% of whole side chains It is 18:1.

" Good Manufacture Practice " and " GMP " is referred to according to 21C.F.R.110 (food on people) and 111 (on food With tonic) condition set up of regulation that proposes, or the area formulation beyond the U.S. equal supervisory system.The U.S. Regulation is promulgated by U.S. Food and drug administration according to Federal Food, medicine and cosmetics bill, is used for the mankind for specification Manufacturer, processing business and the packaging business of the food article of consumption and edible tonic.

" growth " represents the increase of cell size, total cell content and/or cell quality or the weight of individual cells, including Cell weightening caused by fixed carbon source changes into intracellular oil.

" homogenate " represents the biomass crushed by physics.It is such hydrodynamics process to homogenize, its be related to by Particle is separated into smaller and evenly size, forms the dispersion that can be subjected to being processed further.It is homogenized for handling some Food and dairy produce, to improve stability, shelf-life, digestion and taste.

" increased lipid yield " refers to the raising of lipid/oil productivity ratio of culture of microorganism, and its implementation can To be for example, when increasing the dry cell weight in every liter of culture, the percentage for increasing the cell containing lipid and/or increase per unit Between lipid total amount in every liter of culture volume.

" original position " expression " on the spot " or " in its home position ".For example, culture can contain the first of secretion catalyst Second microbial cell type of microalgae cell type and secretion substrate, is being co-cultured wherein the first and second cell types are produced Component of the specified chemical reaction needed for without carrying out the further separation or processing of material occurs for situ.

" lipid " represents to dissolve in non-polar solven (such as ether and hexane) and relative or fully water insoluble any class Molecule.The reason for lipid molecular has these properties is that they are mainly what is be made up of the long hydrocarbon afterbody of hydrophobic in nature.Fat The example of matter includes aliphatic acid (saturation and undersaturated)；Glyceride or glyceride (such as monoglyceride, diglyceride, glycerine three Ester or neutral fat and phosphoglyceride or glycerophosphatide)；With nonglyceride (sphingolipid, tocopherol, tocotrienols, sterol ester, Including cholesterol and steroid hormone, iso-amylene alcohol ester, including terpenoid, fatty alcohol, wax and polyketone).

" lysate " represents the solution containing dissolving cellular content.

" dissolving " is represented the plasma membrane of microorganism and optionally cell wall rupture into being enough to discharge at least some intracellular Content, this is realized often by the machinery or infiltration mechanism that destroy its integrality.

" cracking " is represented the cell membrane of biologic artifact and optionally cell membrane or clasmatosis into being enough to discharge at least Some intracellular contents.

" microalgae " represents the eukaryotic microorganisms organism containing chloroplaset, and it can carry out photosynthesis or can not enter Row photosynthesis.Microalgae is including not energy metabolism fixed carbon source is as the obligate photoautotroph of energy and may exit off fixation The heterotrophic organism that carbon source is individually survived, including photosynthetic obligate heterotroph can not be carried out.Microalgae is included in cell division (for example, volvox, it is two kinds of differences for the unicellular microorganism (such as chlamydomonas) separated soon with sister cell afterwards and microorganism The simple many cells photosynthetic microorganism of cell type)." microalgae " also includes such as chlorella, class chlorella (Parachlorella) With the cell of Du Shi algaes.

" microalgae biomass ", " algal biomass " and " biomass " represents what is produced by the growth and/or breeding of microalgae cell Material.Biomass can contain cell and/or intracellular content and extracellular material.Extracellular material include but It is not limited to the compound secreted by cell.

" microalgae oil " and " algae oil " refer to any lipid composition produced by microalgae cell, including triacylglycerol.

" micronizing " represents the biomass of (or equivalent processing) of homogenizing under high pressure, therefore at least 50% Granularity (median particle size) is no more than 10 μm in the longest dimension or diametrically of the spheroid of their equating volume.Typical case Ground, at least 50% to 90% or more this particle is in the longest dimension or diametrically of the spheroid of their equating volume Less than 5 μm.Under any circumstance, the particle mean size of micronizing biomass is less than the particle mean size of complete microalgae cell.The grain Sub- size is related to those from the result homogenized, and is preferably surveyed after the generation that homogenizes and before the drying Amount, so that avoid may be as the distortion caused by the particle cluster occurred in drying process.Measure some technology (such as laser of granularity Diffraction) detection cluster particle rather than single particle size, and a kind of larger apparent grain can be shown after drying Spend (such as 1-100 μm of particle mean size).Because the shape of these particles is typically approximately spherical, the spheroid of equating volume Diameter and the longest dimension of particle be about identical.

" microorganism " represents any microcosmic unicellular organism.

" nutritional supplement " represents to be intended to supplement diet by providing specific nutrient without being to provide amount of heat.Battalion Foster tonic can contain any one or more of following component: vitamin, mineral matter, herbaceous plant, amino acid, required fat Fat acid and other materials.Nutritional supplement is typically to do piece agent or capsule.It is single to do piece agent or the nutrition benefit of capsule The daily intake of product is typically in no more than 15 grams of level.Nutritional supplement can be provided in can be with food composition (such as acid Milk or " smoothie ") mixing the wafer of mixed type at any time in, to supplement diet, and daily intake level does not surpass typically Cross 25 grams.

" oil " represent by organism (including microalgae, other plant and/or animal) produce any triacylglyceride (or Triglyceride oil).In order to be different from " fat ", unless otherwise indicated, " oil " refers under common room moderate pressure usually The lipid of liquid.For example, " oil " includes the vegetable oil or seed oil for coming from plant, include but is not limited to be derived from following material Oil：Soybean, rapeseed, canola, palm, palm kernel, coconut, corn, olive, sunflower, cottonseed, calyx are away from flower, peanut, Asia Numb shepherd's purse, mustard seeds, cashew nut, oat, lupin, mestha, pot marigold, hemp, coffee, linseed, fibert, the root of Beijing euphorbia, pumpkin, Yuan Sui, camellia, sesame, safflower, paddy rice, tung tree, cocoa, coconut are dry, opium poppy, castor bean, hickory nut, jojoba, leprosy Tree, Queensland nut, Bertholletia excelsa and avocado and combinations thereof.

" osmotic shock " represents that the cell in solution ruptures after osmotic pressure is reduced suddenly, available for induction by cell Cellular component be discharged into solution.

" pasteurization " represents to be intended to slow down the heating process of the microorganism growth in food article.Typically, Pasteur is gone out Bacterium is carried out in high temperature (but less than boiling point) the lower short time.As described herein, pasteurization can be not only reduced in food article The number of undesirable microorganism, but also some enzymes present in food article can be inactivated.

" polysaccharide " and " glycan " represents any carbohydrate for being joined together to form monose by glycosidic bond.Cellulose It is the example for the polysaccharide for constituting certain plants cell membrane.

" port " refers to the opening for allowing material (such as gas, liquid and cell) to flow in or out in bioreactor； Port is generally connected with pipeline.

" mainly encapsulating " represents that more than 50%, typically more than the 75% of the component (such as algae oil) being previously mentioned arrives 90% is insulated in the packing material (can include such as microalgae cell) being previously mentioned.

" mainly complete cell " and " mainly complete biomass " is represented comprising more than 50%, be typically 75%, The cell mass of 90% and more than 98% intact cell." complete " represent to surround the cell of the intracellular members of cell herein The physical continuity of film and/or cell membrane is not destroyed in any way, so that the intracellular members for discharging cell are arrived More than the infiltrative degree of the cell membrane in culture.

" mainly dissolving " represents such cell mass, wherein more than 50%, typically more than 75% to 90% it is thin Born of the same parents have been broken so that the intracellular members of cell are no longer fully enclosed in cell membrane.

" propagation " represents the combination of both growth and breeding.

" breeding " represents to pass through mitosis or the increase of other fissional cell numbers.

" approximate analysis " represents the fat, nitrogen/protein, crude fibre of analysis food material (based on cellulose and lignin Want component), moisture and ash content.Can be calculated by subtracting the given value sum of approximate analysis from 100 (carbohydrate it Difference) soluble-carbohydrate (total dietary fiber and free sugar).

" sonication " is referred to by acoustic wave energy disruption of biological material (such as cell).

It is 2 furan carboxyaldehyde and its derivative of basic structural feature that maintains like that " furfural species ", which is represented,.

" stalk " represents remaining dry stem and leaf after crop harvesting grain.

" being suitable for human consumption " represents that composition can be consumed without unsound influence as diet intake for people, and And important energy intake can be provided because of the material assimilated in the gastrointestinal tract.

" raw food " represents to be not subjected to the composition of heating, but may previously live through heating comprising one or more Component.

" V/V " or " v/v " on volume ratio represents the volume and the volume of said composition of a kind of material in composition The ratio between.For example, mentioning the composition comprising 5%v/v microalgaes oil represents that the 5% of composition volume is made up of (for example, body microalgae oil Product is 100mm³This composition contain 5mm³Microalgae oil), and said composition residual volume (for example, being in this example 95mm³) be made up of other compositions.

On weight than " W/W " or " w/w " represent composition in a kind of material weight and the weight of said composition The ratio between.For example, mentioning the composition comprising 5%w/w microalgae biomass represents that the 5% of composition weight is made up of microalgae biomass (for example, weight is 100mg this composition microalgae biomass containing 5mg), the residuals weight of composition (for example, be in this example 95mg) it is made up of other compositions.

II. the method for preparing microalgae biomass

The invention provides being suitable for the microalgae biomass of human consumption, by this microalgae biomass and comestible composition knot The method of conjunction and the food composition containing this microalgae biomass, the microalgae biomass be rich in nutrient, including lipid and/ Or protein component.The present invention arises partially from such discovery, you can with microalgae biomass be made with floorboard with high oil content and/ Or mix food article the inside, the wherein oil and/or albumen of biomass with excellent function, and by resulting biomass Matter content can completely or partially substitute oils present in conventional food product and/or fat and/or protein.Algae oil can Mainly comprising single saturated oils, with the saturation being commonly found in conventional food product, hydrogenation (trans fats) and how unsaturated Fat is compared, and it is beneficial to health.Algae oil also acts as the healthy and stable edible oil without trans fats.Remaining is micro- Algal biomass can by oil encapsulating, at least up to food article it is ripe untill, thus extend the shelf-life of oil.Kept in cell In the product of whole life, biomass and the natural seen in oil can also protect oil not oxidized, otherwise will Produce offending smell, taste and texture.Biomass also provides some beneficial micro in addition to oil and/or protein Nutrient, such as algae source dietary fiber (both soluble and insoluble carbohydrate), phosphatide, glycoprotein, phytosterol, fertility Phenol, tocotrienols and selenium.

This section summarize first be suitable for the present invention method microalgae type (part A)), chromogenesis formation lack or The method (part B) of the microalgae strain substantially reduced, followed by for breeding the condition of culture (C portion) of biomass, followed by use The concentration step (D parts) for the biomass being processed further in preparation, and the biology prepared in the method in accordance with the invention The chemical composition of matter describes to terminate (E parts).

A.For the microalgae in the method for the present invention

The method according to the invention can use the microalgae of a variety of suitable oil of energy generation and/or lipid and/or protein Species, although the microalgae of suitable oily and/or lipid and/or the protein of naturally-produced high content is preferred.Influence selection For the present invention microalgae Consideration except produce be used for produce the suitable oils, lipid or protein of food article in addition to, Also include：(1) high lipid (or protein) content represented with the percentage of cell weight；(2) it is easy to growth；(3) it is easy to numerous Grow；(4) it is easy to carry out biomass processing；(5) glyceride is composed；(6) without Algae toxins (the dry microalgae life of the following display of example 5 Material and extract from biomass oil or lipid there is no Algae toxins).

In some embodiments, the cell membrane of microalgae must be destroyed during food process (for example, culinary art), to discharge Active component or for digesting, and in these embodiments, with easily animal (such as people or other nonruminants) Intestines and stomach in digest cell membrane microalgae strain be preferred, particularly if when algal biomass will be used to eat raw Tetramune. For the microalgae strain with high-cellulose/hemicellulose level in cell membrane, in general digestibility is reduction. The pepsin digestibility test assessment digestibility of standard can be used.

In specific embodiments, microalgae includes the cell for being by dry weight at least 10% or more oil.In other realities Apply in scheme, microalgae contains by dry weight at least 25%-35% or more oil.In general, in these embodiments, microalgae In oil content it is more, biomass is more nutritious, thus can be trained by dry weight containing at least 40%, at least 50%, The microalgae of 75% or more oil is particularly preferred.For the preferred microalgae in the inventive method can heterotrophic growth (in no light In the case of by sugar) or obligate heterotroph.In food and/or nutriment, not all types of lipids are all It is desirable, because they there may be a bad taste or niff, and shows that stability is poor, or the mouth provided Sense is poor, and these Considerations can also influence the selection of the microalgae in the method for the present invention.

Microalgae from Chlorella is generally useful in the method for the invention.Chlorella is monoplast green alga category , belong to Chlorophyta.Being generally in the shape of for chlorella cells is spherical, about 2 to 10 μm of diameter, and without flagellum.Bead Some kinds of algae are natural heterotrophism.In preferred embodiments, it is protoconch bead for the microalgae in the method for the present invention Algae, chlorella ellipsoidea, atomic chlorella, Chlorella zofinienesi, Chlorella luteoviridis, Kessler Chlorella, heat resistance chlorella (Chlorella sorokiniana), Chlorella fusca var.vacuolata beads Algae, Chlorella cf.minutissima or floating chlorella.Chlorella (particularly Chlorella protothecoides) is to be used for the present invention Method in preferred microorganism because its have high lipid sense into.For the particularly preferred protoconch in the method for the present invention Chlorella species includes those illustrated in following instance.

Other chlorella species being suitable in the method for the present invention include the species being selected from the group, and the group is by following Item composition：anitrata、Antarctica、aureoviridis、candida、capsulate、desiccate、 Ellipsoidea (including 211/42 plant of CCAP), emersonii, fusca (including mutation vacuolata), Glucotropha, infusionum (including mutation actophila and mutation auxenophila), kessleri are (including any UTEX strains 397,2229,398), crawl fan algae (including 37.88 plants of SAG), luteoviridis (including 2203 plants of SAG and become Kind of aureoviridis and lutescens), miniata, (including UTEX plants of cf.minutissima, minutissima 2341), mutabilis, nocturna, ovalis, parva, photophila, pringsheimii, Chlorella protothecoides (including Any UTEX strains 1806,411,264,256,255,250,249,31,29,25 or CCAP 211/8D or CCAP 211/17 and change Kind of acidicola), regularis (including mutation minima and umbricata), reisiglii (including 11/8 plant of CCP), Saccharophila, (including CCAP 211/31,211/32 plant of CCAP and mutation chlorella ellipsoidea), marine chlorella, Simplex, heat resistance chlorella (including 211.40B plants of SAG), sp. (including UTEX strains 2068 and CCAP211/92), Sphaerica, stigmatophora, trebouxioides, vanniellii, chlorella vulgaris (including CCAP 211/11K, 211/80 plant of CCAP and f.tertia and mutation autotrophy chlorella, viridis, chlorella vulgaris, vulgaris F.tertia, vulgaris f.viridis), xanthella and zofingiensis.

The species species of category (and other microalgaes) for the chlorella of the present invention can be by by their genome The same zone of species of some target areas with identifying herein be compared and identify；It is preferred that species be show with herein The species of the homology of the uniformity of the species of identification or at least high level.For example, can be by using primer pair nucleus And/or chloroplast DNA is expanded and is sequenced and realizes special chlorella vulgaris using the method for the appropriate area of genome Class or the identification of strain, in " Identification of Chlorella spp.isolates using such as using Wu ribosomal DNA sequences”(Bot.Bull.Acad.Sin.42:115-121 (2001)) described in method.Ability Field technique personnel can use the Phylogenetic analysis method established, such as ribosomes the Internal Transcribed Spacer (ITS1 and ITS2rDNA), the amplification and sequencing of 23S RNA, 18S rRNA and other conservative genome areas, not only identifies chlorella Species, also identification are suitable for the species of other microalgaes for producing oil and lipid of method disclosed here.On algae Identification and the example of sorting technique, referring to Genetics, 170 (4):1601-10 (2005) and RNA, 11 (4):361-4 (2005)。

Therefore, genomic DNA compares the species for the microalgae that can be suitable for for identification in the present invention.Conservative gene group DNA (DNA for such as, but not limited to encoding 23S rRNA) region can be expanded by microalgae species, and the microalgae species is for example The respective regions of species that can be relevant with preferred microalgae used in this invention on taxology and preferred with those are carried out Compare.The species that selection shows high-level similitude is used in the method for the present invention.Species in Chlorella given below Between the illustrative property example that compares of this DNA sequence dna.In some cases it may be preferred to have for the microalgae in the present invention Encode at least one of sequence listed in 23S rRNA genomic dna sequence and SEQ ID NO 1-23 and 26-27 There is at least 65% nucleotide identity.In other cases, the coding 23S that the microalgae being preferred in the present invention has At least one of rRNA genomic dna sequence and sequence listed in SEQ ID NO 1-23 and 26-27 a variety of have At least 75%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more nucleotide identity.With other compositions Combination is so that before preparing food composition, the Genotyping of food composition and/or algal biomass is also for determining that algae is biological Matter whether from the single above microalgae strain reliable method.

On determining that the sequence of nucleotides or amino acid identity percentage compares, reference is typically used as using a sequence Sequence, cycle tests is compared therewith.In application sequence comparison algorithm, cycle tests and reference sequences input are calculated In machine, subsequence coordinates, and specified sequence algorithm routine parameter are specified if necessary.Then sequence comparison algorithm is according to the journey specified Order parameter calculates percentage of sequence identity of the cycle tests relative to reference sequences.Being compared for the optimal sequence that compares can be with Carried out for example, by following methods：Smith and water man local homology algorithm (Adv.Appl.Math.2:482 (1981))；Needleman and Wunsch homology alignment algorithm (J.Mol.Biol.48:443(1970))；Pearson and Lipman search similarity method (Proc.Nat ' l.Acad.Sci.USA85:2444(1988))；The computer of these algorithms Change and realize (GAP, BESTFIT, FASTA and TFASTA in wisconsin genetics software bag, Genetics Computer Group,575Science Dr.,Madison,WI)；Or visual inspection (generally referring to Ausubel etc., see on).It is adapted to determine that Another algorithm of sequence identity and sequence similarity percentage is BLAST algorithm, and it is described Altschul's etc. J.Mol.Biol.215:In 403-410 (1990).Software for performing BLAST analyses can be by National Center For Biotechnology Information (network addresswww.ncbi.nlm.nih.gov) open acquisition.

In addition to chlorella, other microalgaes category can be also used in the method for the present invention.In preferred embodiments, it is micro- The species of algae is selected from Kessler class ball algae (Parachlorella kessleri), Bai Shi class ball algaes (Parachlorella Beijerinckii), freshwater microalgae (Neochloris oleabundans), Bracteacoccus, including B.grandis, B.cinnabarinas and B.aerius, Bracteococcus sp. or Scenedesmus rebescens.Microalgae species Other non-limitative examples include those species being selected from the group and the species belonged to, and the group is made up of the following：East song shell Algae (Achnanthes orientalis)；Ah Ge Menshi algaes (Agmenellum)；Red tide system Gui algaes (Amphiprora hyaline)；Double eyebrow algaes, including A.coffeiformis, including A.c.linea, A.c.punctata, A.c.taylori, A.c.tenuis、A.c.delicatissima、A.c.delicatissima capitata；Anabena；Algae fibre, including A.falcatus；Gold production algae (Boekelovia hooglandii)；Borodinella；Botryococcus braunii (Botryococcus braunii), including B.sudeticus；Bracteoccocus, including B.aerius, B.grandis, B.cinnabarinas, B.minor and B.medionucleatus；Whip algae；Chaetoceros, including C.gracilis, C.muelleri and C.muelleri subsalsum；Chlorococcum, including C.infusionum；Green shuttle algae；Lan Yinzao；Gold goal Algae；Ball calcium plate algae；The hidden dinoflagellates of Kou Shi；Hidden algae；Small ring algae, including C.cryptica and C.meneghiniana；Du Shi algaes, including D.bardawil、D.bioculata、D.granulate、D.maritime、D.minuta、D.parva、D.peircei、 D.primolecta, D.salina, D.terricola, D.tertiolecta and D.viridis；Only ball algae, including E.viridis；Ellipsoidon；Euglena；Drape over one's shoulders thorn algae；Crisp bar algae, including F.crotonensis；Slimeball algae；Liz algae；Film Born of the same parents algae；Isochrysis galbana, including I.aff.galbana and I.galbana；Squama hole algae；Micractinium pusillum (including UTEX LB 2614)；It is single Pin algae, including M.minutum；Single needle algae；Microballoon algae；Intend Nannochloropsis oculata, including N.salina；Boat-shaped algae, including N.acceptata, N.biskanterae, N.pseudotenelloides, N.pelliculosa and N.saprophila； Neochloris oleabundans；Kidney whip algae；Kidney slit bamboo or chopped wood algae；Common rhombus algae；Rhombus algae, including N.alexandrina, N.communis、N.dissipata、N.frustulum、N.hantzschiana、N.inconspicua、N.intermedia、 N.microcephala, N.pusilla, N.pusilla elliptica, N.pusilla monoensis and N.quadrangular；Ochromonas；Egg capsule algae, including O.parva and O.pusilla；Quiver algae, including O.limnetica and O.subbrevis；Parachlorella, including P.beijerinckii (including 2046 plants of SAG) and P.kessleri (including Any SAG strains 11.80,14.82,21.11H9)；Pascheria, including P.acidophila；Ba Fuzao；Bacteriophage；Xi Zao； Flat algae；Cocolith, including P.carterae and P.dentate；Prototheca, including P.stagnora (including UTEX 327), P.portoricensis and P.moriformis (including UTEX strains 1441,1435,1436,1437,1439)； Pseudochlorella aquatica；Pyramimonas sp；Mulberries algae；Rhodococcus opacus；Sarcinoid chrysophyte；Grid are arranged Algae, including S.armatus and S.rubescens；Schizochytrium limacinum；Water is continuous；Spirulina platensis；Split an algae；Synechococcus；Corner algae； Four algaes, including T.suecica；Wei Shi hailian seaweeds；With Viridiella fridericiana.

In some embodiments, food composition and food ingredients such as algae powder are with coming from one or more selected from following Sequence have at least 90% or 95% 23S rRNA genome sequence uniformity algae：SEQ ID NO:1、SEQ ID NO:2、SEQ ID NO:3、SEQ ID NO:4、SEQ ID NO:5、SEQ ID NO:6、SEQ ID NO:7、SEQ ID NO:8、 SEQ ID NO:9、SEQ ID NO:10、SEQ ID NO:11、SEQ ID NO:12、SEQ ID NO:13、SEQ ID NO:14、 SEQ ID NO:15、SEQ ID NO:16、SEQ ID NO:17、SEQ ID NO:18、SEQ ID NO:19、SEQ ID NO: 20、SEQ ID NO:21、SEQ ID NO:22、SEQ ID NO:23、SEQ ID NO:26 and SEQ ID NO:27.

B.The method for producing the microalgae strain lacked or with the pigment formation substantially reduced

Microalgae (such as chlorella) being capable of photosynthetic growth or heterotrophic growth.When being fixed carbon source and different without light in carbon source It is usually that the microalgae of green has yellow color, marennin formation lacks or substantially reduced when being grown under the conditions of supporting.Marennin is formed Food ingredients can be advantageously acted as by reducing the microalgae of (or shortage).Marennin formation reduces an advantage of the microalgae of (or shortage) It is that the chlorophyll taste of microalgae is reduced.Another advantage that marennin formation reduces the microalgae of (or shortage) is to be used as food ingredients The microalgae added into food material will not produce green color, and green color will not produce attraction to consumer.In heterotrophism bar The reduction of the marennin formation of the microalgae grown under part is temporary transient., being capable of phototrophy and heterotrophism life when gaining phototrophy growth Long microalgae is formed marennin is recovered.In addition, even if marennin is reduced, the microalgae of heterotrophic growth is also yellow color, and this is right It is probably inappropriate for some food applications that consumer expects that food material is white colour or light colour.Therefore, have Profit be produce can heterotrophic growth (therefore marennin formation reduce or lack) and uranidin formation also reduce (therefore for Muted color for food applications) microalgae strain.

Chromogenesis formation lacks or a kind of method of microalgae strain for substantially reducing is by mutagenesis, needed for then screening Phenotype.Several method of mutagenesis are known, and implemented in the art.For example, (Urano etc., J such as Urano Bioscience Bioengineering(2000)v.90(5):Pp.567-569 the ellipse for irradiating and producing using UV) is described The yellow and albino mutant of chlorella.Kamiya(Kamiya,Plant Cell Physiol.(1989)v.30(4):513- 521) chlorella vulgaris (Chlorella vulgaris), 11h (M125) colourless strain are described.

In addition to irradiating mutagenesis by UV, in order to which chromogenesis formation reduces the microalgae of (or shortage), it would however also be possible to employ change Learn mutagenesis.Have proven to the mutagenesis of such as ethyl methane sulfonate (EMS) or N- methyl-N '-nitro-N-nitroguanidine (NTG) etc Agent is effective chemical mutagen to the multiple-microorganism including yeast, fungi, mycobacteria and microalgae.Can also carry out Many wheel mutagenesis, wherein making microalgae contact mutagens (UV or chemical mutagen have both at the same time), then screen required pigment shape Into the phenotype of reduction.Then turn out the colony with required phenotype in flat lining out and separate again, with ensure from a generation to Follow-on mutation is stable, and colony is pure rather than population mixture.

In specific example, with reference to UV mutagenesis and mutagenesis, lacked using the formation of Chlorella protothecoides chromogenesis or The strain of reduction.Chlorella protothecoides is subjected to a wheel mutagenesis with NTG and screen the colony of color change body.Then no table is made The colony for revealing color change is subjected to wheel UV irradiations, and screens color change body again.In one embodiment, separate outstanding Element forms the Chlorella protothecoides strain lacked, and it is Chlorella protothecoides 33-55, according to budapest treaty on October 13rd, 2009 It is deposited in American type culture collection (10801University Boulevard, Manassas, VA 20110- 2209), Patent Deposit number PTA-10397.In another embodiment, the reduced Chlorella protothecoides strain of pigment formation is isolated, its For Chlorella protothecoides 25-32, it is deposited according to budapest treaty on October 13rd, 2009 in American Type Tissue Culture The heart (10801University Boulevard, Manassas, VA20110-2209), Patent Deposit number PTA-10396.

C.The culture medium and condition of culture of microalgae

Microalgae is cultivated in the method according to the invention liquid medium within to breed biomass.In the method for the invention, In the case of without light microalgae species is grown in the culture medium containing fixed carbon and/or fixed nitrogen source.This growth is referred to as Heterotrophic growth.For example for some microalgae species, long-time (such as 10 to 15 days or longer) under conditions of nitrogen is limited Heterotrophic growth causes the lipid that high content is accumulated in cell.

The component that micro-algae culture medium is usually contained for example has fixed carbon source (being discussed below), fixed nitrogen source (such as protein, beans The dregs of rice, yeast extract, corn steep liquor, ammonia (pure or salt form), nitrate anion or nitrate), trace element (for example, respective Form be, for example, ZnCl₂、H₃BO₃、CoCl₂·6H₂O、CuCl₂·2H₂O、MnCl₂·4H₂O and (NH₄)₆Mo₇O₂₄·4H₂In O Zinc, boron, cobalt, copper, manganese and molybdenum), the buffer solution for keeping pH value and phosphate (phosphorus source；Other phosphate can be used).Its He includes salt, such as sodium chloride, for seawater microalgae at component.

In specific example, the culture medium for being adapted to culture Chlorella protothecoides shows culture medium including the moon.This culture medium is adapted to Aseptic culture material, the culture medium (pH value about 6.8) of 1L volumes can be by adding 1g albumen into 1 liter of Bristol culture medium It is prepared by peptone.Bristol culture mediums include 2.94mM NaNO in aqueous₃, 0.17mM CaCl₂·2H₂O, 0.3mM's MgSO₄·7H₂O, 0.43mM, 1.29mM KH₂PO₄With 1.43mM NaCl., can be with for 1.5% agar medium 15g agar is added into 1L solution.Covering solution go forward side by side horizontal high voltage sterilizing, then store at refrigerated temperatures before the use. The other method with the percentages high amounts of oil by dry weight is grown and is multiplied into by Chlorella protothecoides to have been described (referring to example Such as Miao and Wu, J.Biotechnology, 2004,11:85-93, and Miao and Wu, Biosource Technology (2006)97:841-846 (illustrating to obtain the fermentation process of 55% eleocyte dry weight)).Typically, by increase fermentation when Between, while excessive carbon source is provided under conditions of nitrogen limitation can produce high oily algae.

Solid and liquid growth media generally have the source of many acquisitions, are adapted to the defined medium of many microbial strains Preparation explanation be found in for example online http://www.utex.org/ (the nets safeguarded by The University of Texas at Austin Stand, for its algal cultures preservation (UTEX)).For example, various fresh water culture mediums include 1/2,1/3,1/5,1X, 2/3,2X CHEV diatom culture mediums；1:1DYIII/PEA+Gr+；Ag diatom culture mediums；Allen culture mediums；BG11-1 culture mediums；Bold1NV With 3N culture mediums；Grape algae culture medium；Bristol culture mediums；Chu's culture mediums；CR1, CR1-S and CR1+ diatom culture medium； Hot spring red algae culture medium；Blue-green algae culture medium；Band algae culture medium；DYIII culture mediums；Euglena culture medium；HEPES culture mediums；J is trained Support base；Malt medium；MES culture mediums；Modified Bold 3N culture mediums；Modified COMBO culture mediums；N/20 culture mediums；Ochromonas Culture medium；P49 culture mediums；Plain chlamydomonas culture medium；Proteose culture medium；The raw algae culture medium of snow；Soil extract culture medium；Soil Earth water：GR-, GR-/NH4, GR+, GBARR+/NH4, PEA, peat and VT culture mediums；Spirulina culture medium；Tap culture mediums；Altogether Ball algae culture medium；Volvocaceae culture medium；Volvocaceae -3N culture mediums；Volvox culture medium；Volvox powder dextrose culture-medium；Waris Culture medium；With Waris+ soil extract culture mediums.Various Saline Mediums include：1%th, 5% and 1X F/2 culture mediums；1/2、 1X and 2X Erdschreiber ' s culture mediums；1/2nd, 1/3,1/4,1/5,1X, 5/3 and 2X soil+sea water medium；1/4ERD； 2/3 enriching seawater culture medium；20%Allen+80%ERD；Artificial seawater culture medium；BG11-1+.36%NaCl culture mediums； BG11-1+1%NaCl culture mediums；Bold 1NV:Erdshreiber(1:1) with (4:1)；Bristol-NaCl culture mediums；Suede branch Segasso sea water culture medium；1/2 and 1X enriching seawater culture mediums, including ES/10, ES/2 and ES/4；F/2+NH4；LDM culture mediums；Change Property 1X and 2X CHEV；Modified 2X CHEV+ soil；Modifying artificial sea water medium；Unicellular red algae culture medium；With the training of SS diatoms Support base.

Other suitable culture mediums used in the inventive method can easily be given by consulting the URL identified above To identify, or consulting keeps its hetero-organization of culture of microorganism, such as SAG, CCAP or CCALA.SAG refers to brother's Dettingen The algal cultures collection (brother's Dettingen, Germany) of university, CCAP is referred to by Scotland Marine Sciences association (Soviet Union's lattice Orchid, Britain) management algae and protozoan culture collection, CCALA refer to Institute of Zoology (Czech Republic) algae laboratory cultures thing collection.

It is found in suitable for the microorganism according to the inventive method the diverse location and environment of All Around The World.Due to them Separated with other kinds of, and their resulting evolutionary divergences, thus it is likely difficult to or any tool can not possibly be predicted Body microbe species most preferably grow and produced the specific growth medium of oil and/or lipid and/or protein, but this area skill Art personnel are in view of content disclosed herein, appropriate culture medium just can be easily found by carrying out routine experiment. In some cases, certain micro-organisms strain possibly can not be grown on specific somatomedin, because there are some inhibition groups Point, or lack some basic nutritional needs needed for specific microbial strain.Be provided below the various microalgae species of culture with The example of the illustrative methods of the percentages high content lipid of dry cell weight is pressed in accumulation.

Fixed carbon source is the key component of culture medium.For purposes of the invention, suitable fixed carbon source include such as glucose, Fructose, sucrose, galactolipin, xylose, mannose rhamnose, arabinose, NAG, glycerine, floridoside, Portugal Grape uronic acid and/or acetic acid esters.Include mixture, such as glycerine and glucose according to other carbon sources that the present invention is used to cultivate microalgae Mixture, the mixture of glucose and xylose, the mixture and sucrose and the mixture of depolymerization beet pulp of fructose and glucose. Be suitable for cultivate microalgae other carbon sources include black liquor, cornstarch, depolymerizing cellulose material (come from such as maize straw, Beet pulp and switchgrass), lactose, whey, molasses, potato, paddy rice, sorghum, sucrose, beet, sugarcane and wheat.It is a kind of or many The supply concentration for planting carbon source can be at least about 50 μM, at least about 100 μM, at least about 500 μM, at least about 5mM, at least about 50mM At least about 500mM.

Therefore, in various embodiments, the fixed carbon energy source in growth medium includes glycerine and/or 5- And/or 6- carbon sugar, such as glucose, fructose and/or xylose, they may originate from sucrose and/or cellulosic material, including depolymerization fiber Plain material.Multiple strains in the multiple species and species of chlorella can be in the presence of sucrose, depolymerizing cellulose material and glycerine Growth, respectively as described in U.S. Patent Application Publication No. 20090035842,20090011480,20090148918, also joins See PCT Patent Application publication number 2008/151149, combined each of which herein by quoting.

Therefore, in one embodiment of the invention, the use of depolymerizing cellulose biomass is raw material culture microorganism.With Other raw materials of such as cornstarch or the sucrose from sugarcane or beet etc are on the contrary, cellulose biomass is (depolymerization or other Type) be not suitable for human consumption, and it is possible to be used with relatively low cost, this causes it to be for purposes of the invention Particularly advantageous.Microalgae can breed on depolymerizing cellulose material.Cellulosic material generally includes the fibre of 40%-60% dry weights Dimension element；The hemicellulose of 20%-40% dry weights；With the lignin of 10%-30% dry weights.Suitable cellulosic material includes draft Plant and woody energy crop and the residue of crops, i.e. the plant not removed with primary food or fiber product from field Thing part, mainly stem and leaf.Example includes：Agricultural wastes, such as bagasse, rice husk, zein fiber (including stem, leaf, shell and Cob), wheat straw, straw, beet pulp, citrus pulp, orange peel；Forestry waste, the hardwood and cork such as processed from timber is sparselyed fell Material and hardwood and cork residue.Culled wood, such as sawmill's waste material (wood chip, sawdust) and pulp mill's waste material；Municipal waste, such as city The scraps of paper, city culled wood and the urban green space waste material of city's solid waste, grass is cut in such as city；With Wood construction waste material.Other fiber Element includes special cellulosic crops, such as switchgrass, hybridization poplar and Chinese silvergrass, fiber stalk and fiber sorghum.Produced by this class material Raw pentose includes xylose.Example 20 is described under the conditions of heterotrophism to be spread out using the cellulose from maize straw and beet pulp The Chlorella protothecoides that raw sugar success is cultivated.

Some microorganisms can handle cellulosic material and DIRECT UTILIZATION OF CELLULOSE material is as carbon source.However, cellulose Material usually require by processing with increase can and surface area, or for making cellulose be decomposed first, preparation is used as carbon Source is utilized for microorganism.Preparation or pretreatment mode for the cellulosic material of enzymolysis are well known in the art.The side Method is divided into two major classes：(1) cellulosic material is resolved into smaller particle, so as to increase can and surface area；(2) at chemistry Reason cellulosic material can be used for the substrate of enzymolysis to be formed.

Increasing the method for accessible surface product includes steam blasting, and it is related to high-temperature steam come decomposition of cellulose material.This One crosses range request high temperature, therefore some sugar in cellulosic material might have loss, so as to reduce the carbon source for being available for digesting (see, for example, Chahal, D.S. etc., Proceedings of the 2^nd World Congress of Chemical Engineerin g(1981)；With Kaar etc., Biomass and Bioenergy (1998) 14 (3):277-87).Ammonia explosion can Allow cellulosic material to occur explosion at a lower temperature, but it is higher to implement cost, and ammonia potentially interfere with it is follow-up Enzymolysis process is (see, for example, Dale, B.E. etc., Biotechnology and Bioengineering (1982)；12:31- 43).Another blasting technique is directed to use with supercritical carbon dioxide explosion, so as to which cellulosic material is resolved into less fragment (see, for example, Zheng etc., Biotechnology Letters (1995)；17(8):845-850).

It is also as is generally known in the art cellulosic material to be chemically treated to be formed available for the method for the substrate of enzymolysis 's.U.S. Patent number 7,413,882 is described using genetically engineered microorganism, the genetically engineered microorganism secretion Beta-glucosidase into zymotic fluid, and with fermentation liquor treatment cellulosic material with the plain Substance P hydrolysis of reinforcing fiber into glucose. Cellulosic material can also be handled to help follow-up enzymolysis with strong acid and highly basic.U.S. Patent number 3,617,431 describes profit With alkaline digest decomposition of cellulose material.

Chlorella can breed on the culture medium (such as depolymerizing cellulose material) combined containing xylose with glucose, and anticipate Unimaginable to be, when being cultivated in the combination in glucose and xylose, some species are even shown than individually in grape Higher productivity ratio when on sugar or individually being cultivated on xylose.Therefore, some microalgaes can utilize on the contrary unedible raw material (such as cellulosic material (or pretreated cellulosic material) or glycerine) is as carbon source and produces edible oil.This make it possible to by Inedible cellulose and glycerine (are generally not belonging to the food chain of the mankind, this and the corn grape from sugarcane and beet Sugar is opposite with sucrose) edible oil that changes into high nutrition (as a part for the diet of people, can provide nutrient and heat Amount).Therefore, the present invention, which is provided, would prove unsuitable for edible oil, food article and combinations of foods that edible raw material changes into high nutrition The method of thing.

With expressing the organism of secreting type invertase the co-cultures or culture in the culture medium containing invertase Or the exogenous invertase gene of expression (wherein secrete invertase or organism also expresses sucrose transporter) it is micro- Algae can breed on the blackstrap from sugarcane or other sugar sources.It can be shown using this low value, the waste containing sucrose Write the production cost that edible oil is saved on ground.Therefore, microalgae and preparation diet group are cultivated on sucrose material as described herein The method of compound and nutritional supplement is provided the sucrose inversion of low nutrition into high-nutrition oil (oleic acid, DHA, ARA etc.) and containing this Plant the mode of the biomass of oil.

Patent as mentioned above disclose it is middle be described in detail, many different chlorella species and strain are not only in the reagent of purifying On level glycerine and breed very good in the acidifying from biodiesel ester exchange and non-acidizing glycerin by-product product.Expect Less than, the cell division that some chlorella strains are carried out in the presence of glycerine than the cell that is carried out in the presence of glucose Division is faster.Two stage growth course (wherein supplies cell with glycerine first, with the cell density that increases sharply, then supplied Glucose is to accumulate lipid) the seviparous efficiency of production can be improved.

Another raising lipid is directed to use with acetic acid esters as microalgae as the method for the percentage of dry cell weight Raw material.Acetic acid esters is supplied directly to trigger the metabolism point of aliphatic acid synthesis (that is, acetyl-CoA)；Therefore second is provided in culture Acid esters can increase the output of aliphatic acid.Generally, microorganism is cultivated in the presence of enough acetic acid esters, to improve microorganism fat The yield of matter and/or aliphatic acid (specifically, relative to the yield when lacking acetic acid esters).The acetic acid esters of supply is for herein The method of the generation microalgae biomass of offer is a kind of useful component, and the microalgae biomass, which has, accounts for dry cell weight at high proportion Lipid.

In another embodiment, by one or more lipid path enzymes (for example, fatty acid synthetase) it is auxiliary because Culture produces seviparous microalgae to increase lipid yield in the presence of son.Generally, the concentration of co-factor is enough microbial lipids The yield of (for example, aliphatic acid) is increased to exceed the microbial lipids yield in the case of without co-factor.Specifically implementing In scheme, by including secreting the microorganism of co-factor or by adding co-factor into culture medium come to training in culture Support thing and co-factor is provided.Alternatively, microalgae can be engineered and encode the protein for participating in co-factor synthesis to express Allogenic gene.In certain embodiments, suitable co-factor includes the vitamin needed for any lipid path enzyme, for example Biotin or pantothenate.

Compared with low fat quality of biomass, it is favourable material that the high lipids, biological matter in microalgae, which is included in food article, Because so allowing to add less microalgae biomass same amount of lipid is admixed among food composition.So doing is Favourable, because compared with low fat quality of biomass, the healthy oil in high lipid microalgae is added in food article and can not changed Become other attributes, such as texture and taste.The rich lipids, biological matter provided by the inventive method generally has by dry cell weight basis At least 25% lipid.Treatment conditions can be adjusted to improve the percentage by weight of lipid in cell.For example in some embodiment party In case, microalgae is cultivated in the presence of one or more nutriments (such as nitrogen, phosphorus or sulphur) of limiting concentration, while providing The fixation carbon source (such as glucose) of amount.Microbial lipids yield is often increased to exceed and is providing the culture of excess nitrogen by nitrogen limitation Microbial lipids yield in thing.In specific embodiments, lipid yield improve at least about 10%, 50%, 100%, 200% or 500%.Microorganism can be cultivated in the presence of the nutriment of limit amount, incubation time is complete culture period A part or whole culture period.In some embodiments, during whole culture, nutrient concentration is in limiting concentration and non-limit Circulated at least twice between concentration processed.

Under stable growth conditions, Cellular Accumulation is oily but without cell division.In one embodiment of the invention In, by keeping growth conditions to the continuous all components (in addition to fixed nitrogen source) for providing initial growth culture medium of cell.It is logical Cross and supply all nutriments for being initially provided to cell (except fixed nitrogen source) to cell to cultivate microalgae cell, such as by right Cell carries out prolonging prolonged supply, so causes to haggle over high lipid percentage by dry cell weight.

In other embodiments, by all fixed nitrogens exhausted up to long time (such as at least one or two Individual week) fixed carbon source is supplied to cell afterwards to produce high lipids, biological matter.In some embodiments, it is allowed to which cell is solid Determine the presence of carbon source and lack to accumulate oil under fixed nitrogen source more than 20 days.Using condition described here or as is generally known in the art The microalgae of other conditioned growth can include by dry weight at least about 20% lipid, generally comprise by dry weight 35%, 45%th, 55%, 65% and even up to 75% or more lipid.Therefore by the way that cell is maintained at into heterotrophic growth state (its In they consume and carbon and accumulate oil, but without cell division) under the lipid that can improve in microbial lipids production account for cell and do The percentage of weight.

High-protein biological matter in algae is to contain another favourable material in food article.This hair Bright method can also provide the biomass with the protein for accounting for its dry cell weight at least 30%.Growth conditions can adjust with Improve the percentage by weight of protein in cell.In preferred embodiments, in rich nitrogen environment and fixed carbon energy (such as Portugal Grape sugar) or any of above other carbon source excess discussed in the case of cultivate microalgae.The excessive condition of nitrogen is often by microorganism Protein yields be increased to exceed the protein yields of the microorganism cultivated under the conditions of excess nitrogen is not provided.In order that albumen The output highest of matter, cultivates microorganism up to complete culture period preferably in the presence of excess nitrogen.Suitable microalgae nitrogen source can From organic nitrogen source and/or inorganic nitrogen-sourced.

Organic nitrogen source has been used in culture of microorganism since 20th century.Organic nitrogen source (such as corn steep liquor) makes Produce very universal in penicillin used in by mould.It was discovered by researchers that microorganism can be strengthened comprising corn steep liquor in the medium Growth, and cause the raising of product (such as penicillin) yield.Analysis to corn steep liquor it is well established that it is a kind of rich nitrogen source, and And it is also enriched in vitamin (such as vitamin B compound), riboflavin pantothenic acid, nicotinic acid, inositol and nutritional mineral matter (such as calcium, iron, magnesium, phosphorus And potassium) (Ligget and Koffler, Bacteriological Reviews (1948)；12(4):297-311).Organic nitrogen source (such as corn steep liquor) is had been used in yeast, bacterium, fungi and the fermentation medium of other microorganisms.Organic nitrogen source it is non-limiting Example is yeast extract, peptone, corn steep liquor and corn starch.It is preferred that inorganic nitrogen-sourced non-limitative example include (example As) but be not limited to (NH₄)₂SO₄And NH₄OH.In one embodiment, for implement the present invention culture medium comprise only it is inorganic Nitrogen source.In another embodiment, organic nitrogen source is comprised only for implementing the culture medium of the present invention.In another embodiment again In, the culture medium for implementing the present invention is containing organic nitrogen source and inorganic nitrogen-sourced mixture.

In the method for the invention, the microalgae cell in different physiological period stages is entered using bioreactor or fermentation tank Row culture.As an example, the inoculum for producing seviparous microalgae cell is introduced into culture medium；Have before cell starts breeding late Demurrage (lag period).After lag phase, proliferative speed stably increases, into logarithmic phase or exponential phase.Exponential phase, is successively immediately Slowing down for breeding, this is due to that nutrient (such as nitrogen) is reduced, toxicant increase, and quorum sensing mechanism.This slowed down Afterwards, breeding stops, and cell enters stationary phase or reaches stable growth conditions, and this depends on the specific environment for being supplied to cell.For The biomass of rich albumen is obtained, culture is harvested soon after terminating generally during exponential phase or in exponential phase.In order to The biomass of rich lipid is obtained, harvest culture after generally preferably just terminating in exponential phase, by making nitrogen or other key Nutriment (in addition to carbon) exhausts, and forces cell that the carbon source being present in excess is changed into lipid, it is possible thereby to which termination refers to ahead of time The number phase.Condition of culture parameter can be controlled to optimize the combination and/or specific oily production of total oil production, the lipid species produced Amount.

Bioreactor, which is used in heterotrophic growth and propagation method, can provide many advantages.It is understood that ought be herein When in described heterotrophic growth and propagation method using fixed carbon source, cell arranging using light is enable in photosynthetic growing method It is unnecessary to apply.To produce the biomass for food, preferably make in liquid (as an example, such as in suspending nutrient solution) micro- Algae is fermented in large quantities.Such as steel fermentation tank (the present invention different embodiments in using 5000 liters, 10,000 liters, 40,000 Rise and higher volume of steel fermentation tank) etc bioreactor be adapted to very big culture volume.Bioreactor allusion quotation Allow control condition of culture (such as temperature, pH value, oxygen tension and carbon dioxide level) type.For example, bioreactor is typically The port of configurable (for example) connecting pipe, to allow to make gaseous component (as oxygen or nitrogen) can be by nutrient solution bubbling.

Can be by bioreactor configurations into micro-algae propagation and flowing through culture medium on the increased number of whole period Bioreactor.For example in some embodiments, culture medium can be noted before density needed for cell after inoculation reaches Enter bioreactor.In other cases, after inoculum culture, bioreactor is made when culture starts full of training injecting Base is supported, and is not injected into more culture mediums.In other words, in aqueous culture medium by microalgae biomass culture for a period of time, Micro-algae propagation and number increases during this period；However, on the whole described period, substantial amounts of aqueous culture medium without flow through Bioreactor.Therefore in some embodiments, after inoculation, aqueous culture medium is without flow through bioreactor.

It can use and be equipped with such as rotating knife and blade, agitating device, stirring rod, gas-pressurized injection device etc The bioreactor of equipment mixes microalgae culture.Mixing can be continuous either interval.For example implement at some In scheme, the turbulence state of air inlet and culture medium entrance is not kept to breed microalgae, it is described micro- needed for having been carried out The increase of algae number.

As briefly described above, bioreactor is typically equipped with various ports, for example, allow the air content for controlling the culture of microalgae. In order to illustrate this point, the partial volume of bioreactor can be gas rather than liquid, and the air inlet of bioreactor Gas can be drawn into inside bioreactor by mouth.The gas inside bioreactor, which can be advantageously drawn into, includes sky Gas, air/CO₂Mixture, inert gas (such as argon gas) and other gases.Typically by bioreactor configurations into enabling users to Enough control gas enters the speed of bioreactor.As set forth above, it is possible to improve the gas flow into bioreactor to increase The mixing of strong culture.

The turbidity of culture can also be influenceed by improving gas flow.Pass through the horizontal plane air inlet arranged below in aqueous culture medium Port, so makes the gas sparging into bioreactor to the surface of culture, it is possible thereby to realize turbulent flow.It is one or more Air outlet allows gas to enough effusions, thus prevents the accumulation of pressure in bioreactor.It is preferred that air outlet leads to " unidirectional " Valve, " unidirectional " valve prevents contaminative microorganism from entering bioreactor.

Bioreactor described herein, condition of culture and heterotrophic growth and the instantiation of propagation method can be by any Suitable mode is combined, so as to improve the efficiency and/or protein output of microorganism growth.

D. the microalgae concentration after fermenting

According to the microalgae biomass of the microalgae culture generation of method as described above formation in the fermentation medium.For system The standby biomass for making food ingredients composition, is concentrated to biomass, or the harvesting biomass from fermentation medium. From fermentation medium when harvesting microalgae biomass, biomass is mainly comprising the intact cell being suspended in aqueous culture medium. Dehydration is carried out for concentrated biomass.Dehydration or concentration refer to biomass and zymotic fluid or point of other liquid mediums From, therefore be separation of solid and liquid.Therefore during being dehydrated, culture medium is removed from biomass (for example, by biological via holding The filter discharge zymotic fluid of matter), or biomass is otherwise removed from culture medium.Common dewatering include from The heart, filter and use mechanical pressure.These methods be can be used alone, or mode in any combination is used.

Centrifugation relates to the use of centrifugal force separate mixture.In centrifugal process, the component that the density of mixture is larger is remote Centrifugal shaft is migrated, and the low-density fraction of mixture is migrated towards the axle.(that is, centrifugation speed is improved by increasing effective gravitation Degree), the larger material of density (such as solid) is separated with materials of low density (such as liquid), therefore can be separated according to density.It is biological The centrifugation of matter and meat soup or other aqueous solution forms the condensed cream for including microalgae cell.Centrifugation will not remove substantial amounts of intracellular water Point.Actually after centrifugation, still may have in biomass considerable amount of surface moisture or free moisture (for example, 70% with On), therefore it is not considered as that centrifugation is drying steps.

It can also be dehydrated using filtering.The example for being adapted to the filtering of the present invention is tangential flow filtration (TFF), Also referred to as cross-flow filtration.Tangential flow filtration is to make the isolation technics that solid is separated with liquid using membranous system and mobilization force. Geresh, Carb.Polym.50 are can be found on suitable exemplary filter method；183-189 (2002), which depict use MaxCell A/G Technologies 0.45uM hollow fiber filters.Referring also to (such as) and 100kD, 300kD, 1000kD (catalog number (Cat.No.) P2C01MC01), 0.1uM (catalog number (Cat.No.) P2VVPPV01), 0.22uM (catalog number (Cat.No.) P2GVPPV01) and The Millipore that 0.45uM films (catalog number (Cat.No.) P2HVMPV01) are used togetherEquipment.Preferred retentate will not be with notable Level be preferably not adhere to by the product in filter, and retentate on filtering material.Doughnut can also be used Filtration system carries out TFF.The filter that at least about 0.1 micron of aperture is suitable, such as aperture is about 0.12,0.14, 0.16th, 0.18,0.2,0.22,0.45 or at least about 0.65 micron.TFF preferred aperture allows solute in zymotic fluid and broken Piece flows through, but does not allow microbial cell to flow through.

Directly biomass is applied can also influence dehydration, the machinery with separation and fermentation liquid and the mechanical pressure of microbial biomass Pressure is enough to be dehydrated biomass, but does not cause significant cell to dissolve.Can be used belt filter press apply mechanical pressure so that Microbial biomass is dehydrated.Belt filter press is that slurry (for example, being directly derived from the microbial biomass of fermentation tank or bioreactor) is applied Plus the dehydration equipment of mechanical pressure, the coil pipe for the roller that the slurry reduces via diameter passes through between two tension bands.Belt pressure Filter can actually be divided into three areas：Gravity area, wherein discharging free drainage/liquid by porous belts by gravity；Tapered region, its It is middle to prepare the solid applied for pressure；And pressure area, wherein applying adjustable pressure to the solid that gravity is discharged.

After concentration, microalgae biomass can be handled as described below, to produce vacuum-packed cake, algae piece, algae homogenate, algae Powder, algae powder or algae oil.

E. the chemical composition of microalgae biomass

The microalgae biomass produced by cultural method described herein is comprising microalgae oil and/or protein and in fermentation During produced by microorganism or the other components that are mixed into by microorganism from culture medium.

The microalgae with a high proportion of oil/lipid accumulation by dry weight is generated using different cultural methods biological Matter, including use method as known in the art.Microalgae biomass with accumulation oil/lipid at high proportion is useful to the present invention 's.Have up to based on dry cell weight (DCW) in the static culture grown using high ferro (Fe) concentration under autotrophic condition The chlorella vulgaris of 56.6% lipid has been described (Li etc., Bioresource Technology 99 (11):4717-22 (2008)).Having respectively based on DCW for being grown under the conditions of nitrogen hunger in bioreactor and has 60% lipid The Nannochloropsis oculata and Chaetoceros culture of 39.8% lipid also have been described (Rodolfi etc., Biotechnology＆ Bioengineering(2008)).When to photoproduction length and in low nitrogen condition when incising with based on DCW about 30% lipid Edge green alga (Parietochloris incise) culture has been described (Solovchenko etc., Journal of Applied Phycology 20:245-251(2008)).When being grown under the conditions of some heterotrophism in nitrogen hunger, Chlorella protothecoides can be produced Raw based on DCW up to 55% lipid (Miao and Wu, Bioresource Technology 97:841-846(2006)). Through describing other chlorella species (including floating chlorella, heat resistance chlorella and atomic chlorella) when in low nitrogen culture medium Under the conditions of in tank diameter bioreactor grow when accumulated oil (Illman etc., Enzyme for having based on DCW up to 63% and Microbial Technology 27:631-635(2000)).The taller lipid of ratio has been reported based on DCW, bag Include in the Du Shi algae cultures that grow under the conditions of increase NaCl 70% lipid (Takagi etc., Journal of Bioscience and Bioengineering 101(3):223-226 (2006)) and in Botryococcus braunii 75% lipid (Banerjee etc., Critical Reviews in Biotechnology22 (3):245-279(2002)).

Heterotrophic growth cause relatively low content of chlorophyll (with such as open that pond is half congealed or closed photobioreactor system it The phototrophy system of class is compared).The reduction of chlorophyll content would generally improve the organoleptic quality of microalgae, therefore allow to food article The inside blends more microalgae biomass (or the oil thus prepared).It was found that the leaf in heterotrophic growth microalgae (such as chlorella) is green The reduction of cellulose content also reduces the green color in biomass (compared with the microalgae to photoproduction length).Therefore, chlorophyll content Reduce and avoid generally to containing the related bad green color of the microalgae food article to photoproduction length, and allowing to mix in food article Close microalgae biomass or improve the amount for blending microalgae biomass.In at least one embodiment, food article contains chlorophyll The microalgae of the heterotrophic growth of content reduction (compared with the microalgae to photoproduction length).In some embodiments, the leaf of the microalgae powder Chlorophyll contents are less than 5ppm, less than 2ppm or less than 1ppm.

The microalgae biomass rich in oil for being produced by cultural method described herein and being applied to the present invention is included based on DCW At least 10% microalgae oil.In some embodiments, microalgae biomass comprising based on DCW at least 15%, 25% to 35%, 30% to 50%, 50% to 55%, 50% to 65%, 54% to 62%, 56% to 60%, at least 75% or at least 90% it is micro- Algae oil.

The microalgae of biomass described herein oily (or extract of biomass), which can be included, has one or more differences The glyceride of fatty acid ester side chain.Glyceride is made up of the glycerol molecule being esterified with one, two or three fatty acid molecule, institute Different length can be had by stating fatty acid molecule, and can have different saturation degrees.Specific algae oil blend can be with Prepared in the single species of algae, or can be mixed by the biomass (or algae oil) from two or more microalgae species It is combined.

Accordingly it is also possible to control the group of oil by combining the biomass (or oil) from least two different microalgae species Into the i.e. characteristic and ratio of the fatty acid component of glyceride.In some embodiments, at least two of different microalgae species Composed with different glyceride.Different microalgae species together or can be cultivated separately as described herein, preferably in heterotrophism condition Lower culture, so as to produce corresponding oil.Different microalgae species can in the glyceride of cell the difference containing different proportion Fatty acid component.

In certain embodiments, microalgae oil is mainly made up of single unsaturated oils, and such as 18:1 (oleic acid) oil, particularly locates In triglycerides form.In some cases, algae oil is by weight at least 20% single unsaturated oils.In different embodiment party In case, algae oil is by weight or by volume at least 25%, 50%, 75% or more single unsaturated oils (such as 18:1).One In a little embodiments, single unsaturated oils is 18:1、16:1、14:1 or 12:1.In some cases, algae oil be 60% to 75%, The 18 of 64% to 70% or 65% to 69%:1 oil.In some embodiments, microalgae oil is comprising by weight or by volume extremely Few 10%, 20%, 25% or 50% or more esterification oleic acid or esterification alpha-linolenic acid.In at least one embodiment, algae Oil is comprising by weight or by volume less than the 10%, carbon of esterification 22 less than 5%, less than 3%, less than 2% or less than 1% Acid or substantially free of esterification docosahexaenoic acid (DHA (22:6)) (it is especially in triglycerides form).On The example of the microalgae (the hidden dinoflagellates of such as Kou Shi) of high DHA content is produced, reference can be made to United States Patent (USP) 7,252,979,6,812,009 and 6, 372,460.In some embodiments, in microalgae powder oil or the oily lipodogramme of extraction is less than 2%14:0th, 13% to 16%16:0th, 1% to 4%18:0th, 64% to 70%18:1st, 10% to 16%18:2nd, 0.5% to 2.5%18:3 and it is less than 2% carbon chain lengths are 20 or longer oil.

High protein microalgae biomass has been generated using different cultural methods.With the micro- of protein content at high proportion Algal biomass is applied to the present invention.For example, the protein content of different microalgae species have been reported (referring to Becker, Biotechnology Advances(2007)25:207-210, table 1).Have been reported that four whip piece algaes of control Turnover rate in the semicontinuous photoautotrophy culture of (Tetraselmis suecica) influences the protein of each cell to contain Protein (Fabregas etc., the Marine Biotechnology (2001) 3 of amount, up to about 22.8%:256-263).

The microalgae for being produced by cultural method described herein and being applied to the embodiment the present invention relates to high protein is biological Matter typically comprises the protein by dry cell weight basis at least 30%.In some embodiments, microalgae biomass is included by thin Born of the same parents' dry weight meter at least 40%, 50%, 75% or more protein.In some embodiments, microalgae biomass is included by thin Born of the same parents' dry weight meter 30-75% protein or the protein by dry cell weight basis 40%-60%.In some embodiments, microalgae Protein in biomass includes at least 40% digestible crude protein.In other embodiments, the egg in microalgae biomass White matter includes at least 50%, 60%, 70%, 80% or at least 90% digestible crude protein.In some embodiments, microalgae Protein in biomass comprising the digestible crude protein from 40% to 90%, the digestible crude protein from 50% to 80% or from 60% to 75% digestible crude protein.

Microalgae biomass (and the oil thus extracted) can also be comprising being produced or be admixed to life from culture medium by microalgae Other components in material.These other components can exist in different amounts, and this depends on used condition of culture and micro- The species of algae (and if applicable, for reclaiming the extracting method of microalgae oil from biomass).Generally, it is micro- in high protein Chlorophyll content in algal biomass is higher than the chlorophyll content in high lipid microalgae biomass.In some embodiments, The chlorophyll content of microalgae biomass is less than 200ppm or less than 100ppm.Other components may include but be not limited to phosphatide (for example Algae lecithin), carbohydrate, solubility and insoluble fibre, glycoprotein, phytosterol be (for example, cupreol, rape oil steroid Alcohol, stigmasterol, ergosterol and brassicasterol), tocopherol, tocotrienols, carotenoid (such as alpha-carotene, β-Hu Radish element and lycopene), xanthophyll (for example, lutein, luteole, α-kryptoxanthin and beta-cryptoxanthin), protein, polysaccharide (for example, arabinose, mannose, galactolipin, 6- methylgalactoses and glucose) and different organic or inorganic compounds (such as selenium).

In some cases, biomass includes at least 10ppm selenium.In some cases, biomass includes at least 25% W/w polysaccharides.In some cases, biomass includes at least 15%w/w algae glycoproteins.In some cases, biomass Or come from the oil of biomass and be included in total carotinoid between 0-200,0-115 or 50-115mcg/g, and specific 20-70 or 50-60mcg/g total carotinoid composition is lutein in embodiment.In some cases, biomass is included At least 0.5% algae phosphatide.In some cases, biomass or come from biomass oil contain at least 0.10,0.02-0.50, Or 0.05-0.3mg/g total tocotrienols, and 0.05-0.25mg/g is α fertility triolefins in certain embodiments Phenol.In some cases, biomass or come from the oil of biomass and contain in 0.125mg/g to total fertility three between 0.35mg/g Alkene phenol.In some cases, the oil for coming from biomass contains at least 5.0,1-8 or 2-6 or 3-5mg/100g total fertility triolefin Phenols, and 2-6mg/100g is α tocotrienols in certain embodiments.In some cases, biomass is come from Oil contains in 5.0mg/100g to the tocopherol between 10mg/100g.

In some cases, for high-protein biological matter, compared with high lipids, biological matter, other groups of microalgae biomass The composition divided is different.In specific embodiments, high-protein biological matter contains 0.18 to total between 0.79mg/100g Tocopherol, and in specific embodiments, the high-protein biological matter contains about 0.01-0.03mg/g tocotrienols. In some cases, the high-protein biological matter also contains total sterol 1 to 3g/100g, and is in specific embodiments 1.299-2.46g/100g total sterol.The detailed description that tocotrienols and tocopherol in Chlorella protothecoides are constituted is included in In following instance.

In some embodiments, microalgae biomass includes by dry weight 20% to 40% carbohydrate.At other In embodiment, biomass includes by dry weight 25% to 40% or 30% to 35% carbohydrate.Carbohydrate can To be dietary fiber and free sugar (such as sucrose and glucose).In some embodiments, the free sugar in microalgae biomass It is by dry weight 1% to 10%, 2% to 8% or 3% to 6%.In certain embodiments, free sugar components include sucrose.

In some cases, microalgae biomass includes at least 10% Soluble Fiber.In other embodiments, microalgae Biomass includes at least 20% to 25% Soluble Fiber.In some embodiments, microalgae biomass includes at least 30% Insoluble fibre.In other embodiments, microalgae biomass includes at least 50% at least 70% insoluble fibre.Always Dietary fiber is the summation of Soluble Fiber and insoluble fibre.In some embodiments, microalgae biomass is comprising at least 40% total dietary fiber.In other embodiments, microalgae biomass comprising at least 50%, 55%, 60%, 75%, 80%, 90% to 95% total dietary fiber.

In one embodiment, the contents of monosaccharides of the total fiber (total carbohydrates subtract free sugar) be 0.1% to 3% arabinose, 5% to 15% mannose, 15% to 35% galactolipin and 50% to 70% glucose. In other embodiments, the contents of monosaccharides of the total fiber (total carbohydrates subtract free sugar) is about 1% to 1.5% Ah Draw uncle's sugar, about 10% to 12% mannose, about 22% to 28% galactolipin and 55% to 65% glucose.

III. microalgae biomass is processed into finished product food ingredients

The concentration microalgae biomass sheet produced according to the inventive method can be used in food material as finished product food ingredients And without being further modified, or only carry out the modification of bottom line.For example, cake can be vacuum-packed or be freezed. Alternatively, can be by desivac dried biomass, this is a kind of " freeze-drying " process, and wherein biomass is being applied Add in the freeze drying chamber of vacuum and freezed.Cause the water sublimate (primary drying) in biomass to freeze drying chamber applying vacuum With desorption (redrying).However, the invention provides a variety of finished product food ingredients for coming from microalgae, they have by that can apply The enhancing characteristic produced in the processing method of the invention of concentration microalgae biomass.

Mainly complete or homogenate form microalgae biomass is dried be conducive to method described herein and Biomass in composition is processed further or used.Dry refer to remove from mainly complete biomass it is free or Water/the moisture on surface, or remove surface water from the slurry for homogenizing (for example, by micronizing) biomass.According to microalgae Whether biomass is dried, and if if dried, according to the situation of drying means, food article can be assigned Give different texture and flavour.The biomass produced by the microalgae of culture described herein, which is dried, can remove water, and water can It can be the undesirable component in finished product food article or food ingredients.In some cases, dried biomass can help to make The extraction process of microalgae oil is more effective.

In one embodiment, by the microalgae biomass roller drying flakiness form of concentration to produce algae piece, such as originally Described in the part A of section.In another embodiment, the microalgae biomass of concentration is spray-dried or rapid draing (i.e., It is subjected to Optimization of Gas Flow Drying) powder of intact cell is mainly contained to be formed, so that algae powder is produced, in the part B of such as this section It is described.In another embodiment, the microalgae biomass to concentration is micronized and (homogenized) mainly dissolves thin to be formed The homogenate of born of the same parents, itself and then spray-dried or rapid draing produce algae powder, as described in the C portion of this section.In another embodiment party In case, extract oil is to form algae oil from the microalgae biomass of concentration, as described in the D parts of this section.

In some embodiments, be 15% or less by weight after the powder, piece or powder are dried, 10% or less, The water content of 5% or less, 2% to 6% or 3% to 5%.

A.Algae piece

It is applied to the microalgae biomass of concentration as film on the surface of heated roller of rotation, thus from its preparation originally The algae piece of invention.Then dry solid is scraped with knife or blade, obtains small thin slice.U.S. Patent number 6,607,900, which is described, to be made Microalgae biomass is dried in the case of without being centrifuged (concentration) step in advance with drum dryer, such process can To be used in the process of the present invention.

Because biomass is likely to be exposed at high heat during drying, it can be advantageous that before the drying into biomass Add antioxidant.Addition antioxidant can not only protect biomass during drying, and it is micro- to extend the drying of storage The shelf-life of algal biomass.In preferred embodiments, it is biological in the forward direction microalgae of following process (as dried or homogenizing) Antioxidant is added in matter.In the following antioxidant that be discussed in detail and be adapted to use.

In addition, if having between production dehydration microalgae biomass and subsequent process steps for quite a long time, then may Pasteurization advantageously is carried out to biomass before the drying.If had producing between biomass and dried biomass quite The long time, then free fatty can be formed by lipase.The pasteurization of biomass inactivates these lipase, and prevents " soap " taste is formed in resulting dried biomass product.Therefore in one embodiment, the invention provides Microalgae biomass through pasteurize.In another embodiment, the microalgae biomass through pasteurize is algae piece.

B.Algae powder

Using pneumatic conveyer dryer or spray dryer (see, for example, U.S. Patent number 6,372,460), by the microalgae concentrated Biomass prepares the algae powder (or microalgae powder) of the present invention.In spray dryer, the material in liquid suspension is with fine droplets The form of dispersion is ejected into stream of hot air.The material of entrainment is dried rapidly and forms dry powder.In some cases, also may be used To realize powdery texture in final dry matter using pulse combustion dryer.In other cases, it is dry using spraying It is dry to be followed by using the combination of fluidized bed dryer to reach the optimum condition of dried microorganism matter (see, for example, U.S. Patent number 6,255,505).As an alternative, pneumatic conveyer dryer can also be used for the production of algae powder.The thing that pneumatic conveyer dryer will be dried Matter pulls in or is dragged in stream of hot air.When material is dragged into hot-air, moisture is eliminated more rapidly.Then dry is made Separated with humid air, then by humid air recycling for being further dried.

C.Algae powder

By mechanical lysis and homogenize and spray-dried or rapid draing (or use other Air Dried System Dry) turn into the concentration microalgae biomass preparation of powder type algae powder of the invention.The production requirement of algae powder by cell dissolve with Their oil is discharged, and cell membrane and intracellular members are micronized or at least reduced granularity.After homogenizing immediately Or the particle mean size of the particle hereafter measured as soon as possible, preferably more than 10, no more than 25 or no more than 100 microns.At some In embodiment, particle mean size is 1-10,1-15,10-100 or 1-40 μm.In some embodiments, particle mean size is big In 10 μm and up to 100 μm.In some embodiments, particle mean size is 0.1 to 100 μm.

As micronizing discussion in point out, and particularly if by a kind of technology (such as laser diffraction, it measure cluster and It is not individual particle) measure, the particle mean size of particle is preferably measured or hereafter surveyed as early as possible immediately after the generation that homogenizes Amount is (in such as 2 weeks), to avoid potentially causing the distortion of granulometry due to cluster or minimize it.In practice, by Homogenize generation emulsion can generally be stored in refrigerator at least two weeks without material granularity change.Some are used to measure Technology (such as laser diffraction) the measurement cluster of particle rather than the size of individual particle of granularity.Measured cluster of particle has than independent The bigger particle mean size (such as 1-100 microns) of particle.The light microscopy display for the microalgae powder being dispersed in water is single Both only particle and cluster of particle (see Fig. 4).Algae powder is dispersed in water (such as with a blender) with abundant blending but Initial when homogenizing without repeating, these clusters can be destroyed and laser diffraction generally can again be detected and is no more than 10 μm of particle mean size.Software for automating particle size analysis from electron micrograph is commercially available and gone back It can be used for measuring granularity.Here, as elsewhere, particle mean size can be related to any art-recognized average value and measure (such as mean, geometric mean, median or mode).Granularity can be measured by any art-recognized measure, including The longest dimension of particle or the particle diameter of equating volume.Because particle is typically about spherical in shape, these surveys Amount can be substantially identical.

Oil, water and the micronized particle generated after homogenizing is emulsified so that before the drying oil not with dispersion Separation.It is, for example, possible to use the slurry containing cell is aspirated through throttle orifice plate valve to dissolve cell by stress-breaker.Apply High pressure (up to 1500 bars), followed by outlet nozzle instantaneous expansion.Clasmatosis is completed by three kinds of different mechanism：Punching The valve, the high liquid shear in aperture and the unexpected pressure drop after electric discharge are hit, causes the explosion of cell.Methods described is released Put intracellular molecule.Niro (Niro Soavi GEA) homogenizers (or any other high pressure homogenisers) can be used thin Born of the same parents are processed into the particle of mainly 0.2 to 5 micrometer length.Microalgae biomass is handled under high pressure (about 1000 bar) typically molten Cell of the solution more than 90%, and granularity is decreased below 5 microns.

Alternatively, ball mill can be used.In ball mill, cell in suspension by small abrasive particle (such as Bead) agitation.Cell is ruptured based on the grinding between shearing force, bead and with the collision of bead.These beads break cell Split to discharge cellular content.In one embodiment, it is broken using Dyno-mill ECM Ultra (CB Mills) ball mill Broken microalgae biomass is simultaneously formed as stable emulsion.Such as (example can also be used as by means of blending by shearing force come smudge cells Son, as with homogenizer or waying blenden), French cell press or even centrifugation (if cell membrane is weak) come Smudge cells.Suitable ball mill including ball size and blade details is described in U.S. Patent number 5,330,913.

The direct products of homogenizing are the slurries of the size particulate smaller than the initial cell being suspended in oil and water.The particulate Represent cell debris.Oil and water are discharged by cell.Aqueous culture medium containing cell before other water may be from homogenizing. The form of particle is preferably the homogenate of micronizing.If shelving, some less particles can be coalesced.However, it is possible to logical Cross with crystallite stabilizer (such as microcrystalline cellulose) seeding to preserve short grained homogeneous dispersion.

For shape algae powder, slurry is spray-dried or rapid draing, remove water and leave containing cell debris and The dry powder-shaped material of oil.Although the oil content of powder (that is, as the smudge cells of flour) can be for dry powder by weight At least 10%, 25% or 50% of meter, the powder, which can have, to be dried rather than greasy sensation and outward appearance are (for example, without can The oil seen), it can also be flowed freely during shake.Can also add different glidants (including product derived from silica, it is such as heavy Silica, aerosil (fumed silica), calcium silicates and the lagoriolite in shallow lake).By these materials applications in Higher fatty acid, hygroscopicity or cohesive powders can prevent from luming after drying and in packaging, promote the free-flowing of dried powder, and And adhesion, accumulation and oxidation of the material in dryer surface can be reduced.Under the highest level that FDA is specified, it is all this All it is approved for food a bit.The water content or moisture of powder are typically less than 10%, 5%, 3% by weight after drying Or 1%.Other driers (such as pneumatic conveyer dryer or pulse combustion dryer) can also be used to produce algae powder.

The oil content of algae powder can be varied from according to the oily percentage of algal biomass.It can be given birth to by the algae of different oil content Material produces algae powder.In certain embodiments, algae powder is produced by the algal biomass of identical oil content.In other embodiment party In case, algae powder is produced by the algal biomass of different oil content.In the latter case, the algae life of different oil content can be combined Material, then carries out homogenization step.In other embodiments, the algae powder of different oil content is produced first, then with difference Ratio be blended into together, to obtain the algae powder product containing oil content needed for final.In a further embodiment, can be with The algal biomass of different lipodogrammes is combined, then homogenizes to produce algae powder.In another embodiment, give birth to first The algae powder of different lipodogrammes is produced, is then blended into different proportions together, to obtain the algae containing final required lipodogramme Powder product.

The algae powder of the present invention is prepared suitable for extensive food.Due to the original of oil content, fiber content and micronized particle Cause, algae powder is multi-functional food ingredients.Algae powder can be used for baked goods, speed bread processed, yeast containing dough product, egg products, tune Taste product, sauce, nutritious drink, algae milk, pasta and the product without seitan.Product without seitan can use algae powder Made with another product without seitan, such as three-coloured amaranth powder, kudzuvine root starch, buckwheat, ground rice, olecranon bean powder, cornflakes, maize Powder, millet powder, dehydrated potato powder, farina, quinoa powder, sorghum flour, soy meal, bean powder, legume flour, cassava (cassava) powder, eragrosits abyssinica powder, globe artichoke powder, almond powder, acorn flour, coconut powder, chest nut powder, corn flour and Taro meal.Algae powder without seitan in food article of the manufacture without seitan is being useful into subassembly with other, is such as dried Roast food (cake, cookies, Brownie and pie product (such as muffin)), bread, cereal preparation, crispbread, Yi Jiyi Big profit wheaten food.More use algae powder prepare the detailed description of these and other food article in the following example.

Algae powder, which can be used in baked goods, replaces normal fat source (for example, oil, butter or margarine) and eggs.Dry Roasting food and the product without seitan have first-class moisture and bits granular texture (cumb structure), the bits grain Shape structure and what no difference of the conventional baked goods being made of butter and eggs.Due to first-class moisture, These baked goods have the longer shelf-life compared with the conventional baked goods produced without algae powder, and can keep more long Their original texture.

The water activity (Aw) of food can be the indicant preserved the shelf-life in the food article of preparation.Water is lived Degree (scope is from 0 to 1) is that water in being present in food article how to be actively engaged in chemically or physically reacting one measures.Generation The water activity of some common foods of Table A w spectrums is：Fresh fruit/meat/milk (1.0-0.95)；Cheese (0.95-0.90)；It is artificial Butter (0.9-0.85)；Nut (0.75-0.65)；Honey (0.65-0.60)；Cured meat (0.85-0.80)；Jam (0.8- 7.5)；Pasta (0.5)；Cookies (0.3)；And dry vegetalbe/crispbread (0.2).It is most when water activity is less than 0.91 Number bacterium will not grow.During less than 0.80, most of moulds can not grow, and less than 0.60 when do not have microorganism growth be can Can.By measuring water activity, it is possible to the potential rotten source of prediction.Water activity can also be it is determined that enzyme and dimension in food Played an important role in the activity of raw element, they have material impact to food colors, taste and fragrance.

Algae powder can function as the fat gain agent being used in smoothie, sauce or flavouring.The uniqueness of algae powder composition It is that it can transmit the sensible quality and mouthfeel for being comparable to the food article with higher fat.This also confirms that algae powder is made For the ability of texture modifying agent.Flavouring, sauce and the beverage being made of algae powder have close to conventional more higher fatty acid recipe Rheological characteristic and opacity, although these food articles contain approximately half of fats/oils level.Algae powder is also first-class breast Agent, it is adaptable to need the food formulations (such as preparing sauce, flavouring and soup) of denseness, opacity and viscosity.Find in addition Lipodogramme in algae powder of the present invention described herein does not contain trans fats, and with butter or margarine (or other animals Fat) compare, the wholesome unsaturated fat with high level.Therefore the product being made of algae powder can have relatively low Fat content (have more healthy fat), without sacrificing using the normal fat source same food system that routinely recipe is made The mouthfeel and organoleptic quality of product.A kind of use algae powder of one sense organ team for evaluation (has identical fat content low as one kind Adipose versus) make food article.It is also tested for a kind of non-fat control and full adipose versus.Fig. 6 demonstrates the algae powder Fat gain quality.The algae powder product is pursued similar to full adipose versus, particularly in denseness, glutinous mouthfeel (mouthcoating) And in terms of its modality of sensation how to be mixed with saliva.

Algae powder can also be added in the powdery being typically used in the reception of catering service environment or liquid eggs.One The composition of kind of powder egg products and algae powder is a kind of powder in itself, it can with a kind of edible liquid or other edibles into And then subassembly, typically cooked, to form a kind of food article.In some embodiments, the algae powder can be with A kind of combination of flowing product, then the flowing product will be sprayed dry with formed a kind of powdered food composition (such as powder egg, Powdered sauce mixes, powdered soup mixture etc.).In these cases, it is advantageous that after homogenizing but The algae powder (so it is a kind of slurry or dispersion) is combined with a kind of flowing product before drying, the group is then spray-dried Compound, forms powdered food composition.Compared with the dried forms of the two components are mixed, this co-dried mistake Journey will increase the uniformity of the powdered food composition.The addition of algae powder improve the outward appearances of powdery and liquid eggs, texture and Mouthfeel, and improved outward appearance, texture and mouthfeel can also be continued over time, or even the egg made is maintained at steaming When on vapour table.Specific formula and sense organ group result are described in following instance.

By the way that algae powder is combined with one or more comestible compositions and liquid (such as water), algae powder can be used for preparing what is restored The food article of the food article recovery can be a kind of beverage, flavouring (such as salad dressing juice), sauce (such as cheese sauce), Or a kind of intermediate product (such as the dough/pasta that then can be baked).In some embodiments, the food article of the recovery is then made It is subjected to shearing force (such as pressure breaking or homogenize).This has the effect for reducing algae powder particle diameter in the final product, because should The powder of floorboard with high oil content can cause reunion during recuperation.In the food article of recovery, algae powder footpath preferably is average 1 To 15 microns.

D.Algae oil

On the one hand, the present invention relates to prepare the side of algae oil by harvesting algae oil from algal biomass under gmp conditions Method, the algal biomass includes by dry weight at least 15% oil, and wherein algae oil is 18 more than 50%:1 lipid.In certain situation Under, algal biomass includes the mixture of at least two different microalgae species.In some cases, at least two different microalgae kind Class is separated culture.In at least one embodiment, at least two different microalgae species have different glyceride spectrums. In some cases, algal biomass is derived from the algae of heterotrophic growth.In some cases, it is all or at least two different micro- Algae species is containing by dry weight at least 15% oil.

On the one hand, the present invention relates to the method for making food composition, including the algae oil and one kind that frustule will be come from Or the combination of various other comestible compositions is to form food ingredients composition, the frustule contain by dry weight at least 10% or At least 15% oil.In some cases, this method prepares algae oil under the conditions of being additionally included in GMP.

Can be from for food article (in addition to other application)) dissolving biomass in separate algae oil.After oil extract Remaining algal biomass is referred to as degreasing meals.By dry weight or stereometer, the oil that degreasing meals contain is than microalgae before extraction Contained oil is few.Typically, 50%-90% oil is extracted, so that degreasing meals are before extraction containing (for example) 10%-50% oil content.However, biomass still has height in terms of the content of protein and other compositions discussed above Nutritive value.Therefore, degreasing meals can be used in animal feed or human foods application.

In some embodiments of this method, algae oil at least 50%w/w is oleic acid, and contains the DHA less than 5%. In some embodiments of this method, algae oil at least 50%w/w is oleic acid, and contains the DHA less than 0.5%.In the party In some embodiments of method, algae oil at least 50%w/w is oleic acid, and sweet more than 18 containing the carbon chain lengths less than 5% Grease.In some cases, the frustule for producing algae oil includes the mixture of the cell from least two different microalgae species. In some cases, at least two different microalgae species are separated cultures.In at least one embodiment, at least two Different microalgae species has different glyceride spectrums.In some cases, frustule is cultivated under the conditions of heterotrophism.One In the case of a little, at least two different microalgae species are all containing by dry weight at least 10% or at least 15% oil.

On the one hand, the present invention relates to prepare under gmp conditions containing at least 50% single unsaturated oils and containing few In 1%DHA algae oil.In some cases, single unsaturated oils is 18:1 lipid.In some cases, algae oil is packaged in use In the capsule of delivering unit dose gauging.In some cases, algae oil is derived from the mixture of at least two different microalgae species. In some cases, at least two different microalgae species are separated cultures.In at least one embodiment, at least two Different microalgae species has different glyceride spectrums.In some cases, algae oil is derived from the algae cultivated under the conditions of heterotrophism Cell.In some embodiments, the algae oil contains the entitled " chemical group of microalgae biomass as discussed in previous section Into " same composition.

On the one hand, the present invention relates to include 18 more than 60%:1 and at least oil of 0.20mg/g tocotrienols.

On the one hand, the present invention relates to include 18 more than 60%:1 ester (being preferably triglycerides) and at least 0.20mg/g The fatty acid alkyl esters composition of tocotrienols.

The algae oil of the present invention is prepared by the microalgae biomass cleaned concentrated by extracting.Cell in biomass is existed Dissolved before extraction.Optionally, (drying, lyophilized etc.) microbial biomass can also be dried before dissolving (clasmatosis).As Other selection, upon completion of the fermentation, can dissolve cell, and without being separated from part or all of zymotic fluid.For example, working as When cell is dissolved, the ratio between cell and extracellular liquid can be less than 1:1v:v.

The microalgae containing lipid can be dissolved to produce lysate.As detailed herein, any convenient means can be passed through The step of completion dissolving microorganism (also referred to as cell dissolving), including thermal induction dissolve, add alkali, acid adding, use enzyme (such as albumen Enzyme) and polysaccharide degrading enzyme (such as amylase), dissolved using ultrasound, the dissolving based on mechanical pressure and using osmotic shock.It is molten Each for solving these methods of microorganism can be applied in combination by single method or simultaneously or sequentially.Can be by aobvious Degree of the micro mirror analysis and observation to clasmatosis.The cell more than 70% is generally observed using one or more above-mentioned methods Rupture.Preferably, cell rupture is more than 80%, more preferably above 90%, most preferably from about 100%.

The lipid and oil produced according to the present invention by microalgae can be collected by way of extraction.In some cases, may be used To be extracted using organic solvent or oil, or solvent-free extraction procedure can be used.

Extracted for the organic solvent of microalgae oil, organic solvent preferably is hexane.Typically, organic solvent is directly added It is added in lysate, without isolating lysate component in advance.In one embodiment, make by one or more State lysate and organic solvent exposure enough a period of times of method generation so that lipid composition forms molten with organic solvent Liquid.In some cases, then can further refined soln to collect specific required lipid composition.Then mixing can be filtered Thing simultaneously removes hexane by (such as) rotary evaporation.Hexane extraction process is well known in the art.Referring to Frenz etc., Enzyme Microb.Technol.,11:717(1989)。

Miao and Wu describe the scheme that microalgae lipid is reclaimed from the culture of Chlorella protothecoides, wherein being received by centrifuging Cell is obtained, water washing and is freeze-dried with distillation.Resulting cell powder is crushed in mortar, is then carried with n-hexane Take.Miao and Wu, Biosource Technology97:841-846(2006).

In some cases, can be using liquefaction (see, for example, Sawayama etc., Biomass and Bioenergy 17:33-39 (1999) and Inoue etc., Biomass Bioenergy 6 (4):269-274(1993))；Fluid (see, for example, Minowa etc., Fuel 74 (12):1735-1738(1995))；Or supercritical CO₂Extract (see, for example, Mendes etc., Inorganica Chimica Acta 356:328-334 (2003)) mode extract microalgae oil.Pass through supercritical CO₂Extract An example for carrying out extract oil is described as follows.Via supercritical CO₂The extraction algae oil of extraction contains all biological from the algae The sterol and carotenoid of matter, and as the extraction process effect and natively be free of phosphatide.Residual from the process Thing substantially comprises degreasing algal biomass (without oil), but still remains the albumen and carbon hydrate of the preextraction algal biomass Thing.Therefore, the algal biomass of the degreasing of the residual is the suitable raw material for producing algae protein concentrate/isolate, and also It is used as a kind of source of dietary fiber.

Oil extract includes directly adding oil into lysate without separation lysate component in advance.After addition oil, Lysate is voluntarily or because the reasons such as centrifugation are segregated into different layers.The order that the layer is reduced by density may include：Weight Solid precipitation, aqueous phase, emulsion phase and oil phase.Emulsion is mutually the emulsion of lipid and aqueous phase.According to what is added relative to lysate The percentage (w/w or v/v) of oil, centrifugal force (if any), the volume and other factors of aqueous culture medium, may have One or both of emulsion phase and oil phase.Cultivation or the place of enough a period of times is mutually carried out to cell lysates or emulsion with oil Reason so that the Lipid dissolution produced by microorganism forms heterogeneous mixture in oil.

In various embodiments, the oil for extraction process is the oil being selected from the group, and the group is made up of the following：Greatly Beans, rapeseed, canola, palm, palm kernel, coconut, corn, waste vegetable oil, Chinese tallow tree, olive, sunflower, cottonseed, chicken Fat, tallow, lard, microalgae, large-scale algae, calyx are away from flower, flax, peanut, choice white grease (lard), False flax, mustard seed, waist Really, oat, lupin, mestha, pot marigold, hemp, coffee, linseed, hazelnut, the root of Beijing euphorbia, pumpkin, coriandrum, camellia, sesame Fiber crops, safflower, rice, tung tree, cocoa, copra, opium poppy, castor bean beans, hickory nut, jojoba, manioca, Australia are hard Really, Bertholletia excelsa and avocado.The oil mass added into lysate is typically greater than (presses with the 5% of the oily lysate combined V/v and/or w/w meterings).Therefore, the oily preferred v/v or w/w exceed cell lysates 5%, 10%, 20%, 25%th, 50%, 70%, 90% or at least 95% for cell lysates.

Lipid can also be extracted from lysate by solvent-free extraction procedure, by hot-cold lysis product, substantially Without using any organic solvent.Sonication can also be used, particularly if temperature is between room temperature and 65 DEG C.It is this Lysate can be layered through centrifugation or sedimentation, wherein one layer is water:Lipid layer.Other layers can include solid precipitation, water layer And lipid layer.By freeze thawing or it otherwise can cool down emulsion and lipid is extracted from emulsion layer.In this approach, no Need to add any organic solvent or oil.If adding any solvent or oil, it can be less than the 5%v/v of lysate Or w/w.

IV. microalgae biomass or material therefrom are combined with other food ingredients

On the one hand, the present invention relates to include at least 0.1%w/w microalgae biomass and other one or more edibles The food composition of composition, wherein algal biomass include by dry weight at least 10% oil, and the oil of optional wherein at least 90% is Glyceride.In some embodiments, algal biomass contains the oil of at least 25%, 40%, 50% or 60% by dry weight.One In the case of a little, algal biomass contains the oil of 10%-90%, 25%-75%, 40%-75% or 50%-70% by dry weight, appoints The oil for selecting wherein at least 90% is glyceride.In at least one embodiment, by weight at least 50% oil is single insatiable hunger And glycerin fatty oil.In some cases, by weight at least 50% oil is the 18 of glyceride form:1 lipid.In certain situation Under, the oil by weight less than 5% is docosahexaenoic acid (DHA) (22:6).In at least one embodiment, by weight Oil of the meter less than 1% is DHA.Algae lipid content with low-level polyunsaturated fatty acid (PUFA) is preferred, to ensure The chemical stability of biomass.In preferred embodiments, algal biomass is grown under the conditions of heterotrophism, and marennin is formed Reduce.In other embodiments, microalgae is the color change body of pigment formation shortage or reduction.

On the other hand, the present invention relates to include at least 0.1%w/w algal biomass and other one or more edibles The food composition of composition, wherein algal biomass are comprising at least 30% protein by dry weight, by dry weight at least 40% Protein, by dry weight at least 45% protein, by dry weight at least 50% protein, by dry weight at least 55% egg White matter, by dry weight at least 60% protein or by dry weight at least 75% protein.In some cases, microalgae is biological Matter contains 30%-75% by dry weight or 40%-60% protein.In some embodiments, at least 40% crude protein Be digestible, at least 50% crude protein be digestible, at least 60% crude protein be it is digestible, at least 70% it is thick Albumen is that digestible, at least 80% crude protein is digestible or at least 90% crude protein is digestible.At some In the case of, algal biomass grows under the conditions of heterotrophism.In at least one embodiment, algal biomass is to be full of bar in nitrogen Grown under part.In other embodiments, microalgae is the color change body of pigment formation shortage or reduction.

In some cases, algal biomass includes mainly complete cell.In some embodiments, food composition Comprising oil, the oil is primarily or entirely wrapped in the intracellular of biomass.In some cases, food composition is comprising main The complete microalgae cell in ground.In some cases, algae oil is mainly wrapped in the cell of biomass.In other cases, it is raw Cell (for example, homogenate) of the material comprising mainly dissolving.As described above, this homogenate can be provided as slurry, thin slice, powder or Powder.

In some embodiments of food composition, algal biomass is also comprising at least 10ppm selenium.In certain situation Under, biomass is further comprising at least 15%w/w polysaccharides.In some cases, biomass further includes at least 5%w/ W algae glycoproteins.In some cases, biomass includes 0 total carotinoid for arriving 115mcg/g.In some cases, it is biological Matter includes at least 0.5%w/w algae phosphatide.In all cases, as just mentioned, these components are real cells Component rather than extracellular component.

In some cases, the algal biomass of food composition contains the component for possessing antioxidant properties.Strong anti-oxidation Agent characteristic is attributable to a variety of antioxidants present in algal biomass, including but not limited to carotenoid, essential minerals (such as zinc, copper, magnesium, calcium and manganese).Algal biomass is also shown containing other antioxidants, such as tocotrienols and tocopherol.This The member of a little vitamin E series is important antioxidant, and with other wholesome benefits, is such as caused for apoplexy The protective effect of damage, the growth for reversing obstruction of artery, suppressing breast cancer and prostate gland cancer cell, reduction cholesterol levels, Reduce the risk of type ii diabetes and the protective effect for glaucomatous damage.The natural origin of tocotrienols and tocopherol can See the oil and olive oil produced by palm, sunflower, corn and soybean, but the tocotrienols of composition provided herein Level is significantly higher than hitherto known material.

In some cases, food composition of the invention contains algae oil, the algae oil comprising at least 5mg/100g, at least 7mg/100g or at least 8mg/100g total tocopherol.In some cases, food composition of the invention contains algae oil, described Algae oil includes at least 0.15mg/g, at least 0.20mg/g or at least 0.25mg/g total tocotrienols.

In the specific embodiment of above-mentioned composition and/or method, microalgae can produce carotenoid.In some realities Apply in scheme, the carotenoid produced by microalgae can extract (that is, oil or lipid jointly with the lipid or oil produced by microalgae Tocopherols will be contained).In some embodiments, the carotenoid produced by microalgae is xanthophyll.In some embodiments In, the carotenoid produced by microalgae is carrotene.In some embodiments, the carotenoids produced by microalgae is recklessly The mixture of radish element and xanthophyll.In various embodiments, the carotenoid produced by microalgae is selected from comprising at least one The carotenoid of the following group, the group is made up of the following：Astaxanthin, lutein, luteole, alpha-carotene, trans β-Hu Radish element, Cis-β-Carotene, lycopene and their any combination.Oily carotenoid from Chlorella protothecoides The non-limitative example of spectrum is included in the following example.

In some embodiments of food composition, algal biomass is derived under the conditions of Good Manufacture Practice (GMP) Culture and the algae dried.In some cases, algal biomass is combined with other one or more comestible compositions, described one Plant or various other comestible compositions include but is not limited to cereal, water fruits and vegetables, protein, lipid, herbaceous plant and/or perfume (or spice) Expect composition.In some cases, food composition is salad-dressing, egg products, baked goods, bread, food bars, Italy Wheaten food, sauce, soup drink, beverage, frozen dessert, butter smear food.In specific embodiments, food composition is not Ball or powder.In some cases, it is at least 50g or at least 100g according to the weight of the food ingredients composition of the present invention.

Biomass can be combined to prepare food article with other one or more comestible compositions.Biomass may be from The algal biomass of single algae source (for example, strain) or a variety of sources (for example, not homophyletic).Biomass can be from single algae kind Class, but composed with different compositions.For example, the high microalgae of the oil content microalgae high with protein content can be total to by manufacturer It is mixed, until the definite oil and protein content in finished product food article are desired.It can be implemented by food production business Combination is used with making finished product for retail or food reception.As other selection, manufacturer can regard algal biomass as production Product are sold, and algal biomass (for example) can be incorporated into food article by consumer by changing conventional recipe.No matter which The situation of kind, algal biomass is generally all used for substituting all or part of oil, fat, the eggs used in many conventional food products Deng.

On the one hand, the present invention relates to include at least 0.1%w/w microalgae biomass and other one or more edibles The manner of formulation of the food composition of composition, wherein microalgae biomass is, will contain at least algae of 40% protein by dry weight Biomass is blended with the algal biomass containing 40% lipid by dry weight, obtains the protein and fat of required percentage by dry weight The blend of matter.In some embodiments, biomass is from the strain of identical algae.As other selection, it will contain and press dry weight The algal biomass of at least 40% lipid (lipid contained is DHA less than 1%) is counted with containing at least 20% lipid by dry weight The algal biomass blending of (at least the 5% of the lipid contained be DHA), obtain altogether containing at least 10% lipid by dry weight and The blend of 1%DHA dry biomass.

On the one hand, the present invention relates to the method for preparing microalgae biomass under gmp conditions, mode is trained by drying algae Thing is supported, obtains including the algal biomass of by dry weight at least 15% oil, wherein algae oil is greater than 50% single unsaturated lipids.

On the one hand, the present invention relates to the algal biomass containing by dry weight at least 15% oil being made under gmp conditions, Wherein algae oil be greater than 50% 18:1 lipid.On the one hand, the present invention relates to be made under gmp conditions containing by dry weight The algal biomass of at least 40% oil.On the one hand, the present invention relates to be made under gmp conditions containing by dry weight at least 55% The algal biomass of oil.In some cases, packed algal biomass as tablet, the biomass for delivering unit dose. Under certain situation, algal biomass is packaged into label or otherwise with label, the label provides algae is biological The guidance that matter is combined with other comestible compositions.

On the one hand, the present invention relates to by microalgae biomass as described above and/or material therefrom and following institute Other at least one finished product food ingredients stated combine the method to form food composition or food material.In different embodiment party In case, the food composition formed by the method for the present invention includes egg products (powdery or liquid), pasta, seasoning production Product, yolk jam product, cake product, bread product, energy stick, dairy produce, juice product, smearing food or smoothie.One In the case of a little, food composition is not ball or powder.In various embodiments, the weight of food composition is at least 10g, extremely Few 25g, at least 50g, at least 100g, at least 250g or at least 500g or heavier.In some embodiments, it is biological by microalgae The food composition that the combination of matter and/or product therefrom is formed is raw product.In other cases, food composition It is ripe product.

In other cases, food composition is ripe product.In some cases, except belonging to the oil of microalgae biomass Outside, food composition contains the oily or fat by weight less than 25%.When hydrogenated vegetable oil, saturation triglycerides is made The fat (TAG or trans fats) of form, implements as making smearing food (such as margarine).Included in algal biomass Fat there is no any trans fats.In some cases, in addition to the oil for belonging to biomass, food composition contains 10% oily or fat is less than by weight.In at least one embodiment, in addition to the oil for belonging to the biomass, food Composition not oil-containing or fat.In some cases, food composition is free of the oil in addition to the oil for belonging to biomass.One In the case of a little, food composition is free of egg or egg products.

On the one hand, the present invention relates to the method for making food composition, wherein with containing by weight at least 10% The algal biomass of oil completely or partially substitutes the fatty or oil in conventional food product.In one embodiment, this method Determine that the algae for replacement is biological including the oily or fatty amount in the ratio and conventional food product with algae oil in biomass The amount of matter, oil by algal biomass and other at least one comestible compositions and less than the amount included in conventional food product or Fat composition, so as to form food composition.In some cases, the amount of the algal biomass combined with least one other compositions It is 1-4 times of oil and/or fat mass in conventional food product or volume.

In some embodiments, the above method also includes providing and is used for containing other at least one comestible compositions and oil Or the recipe of the conventional food product of fat composition, and by the algal biomass of 1-4 times of quality or volume with reaching conventional food product In fatty or oily quality or volume at least one other comestible compositions combination.In some cases, this method is also wrapped Include and prepare algal biomass under gmp conditions.

In some cases, the food composition bag formed by the combination of microalgae biomass and/or product therefrom Containing the micro- of at least 0.1%, at least 0.5%, at least 1%, at least 5%, at least 10%, at least 25% or at least 50%w/w or v/v Algal biomass or microalgae oil.In some embodiments, the food composition formed as described herein comprising at least 2%, at least 5%th, at least 10%, at least 25%, at least 50%, at least 75%, at least 90% or at least 95%w/w microalgae biomass or by Its obtained product.In some cases food composition comprising by weight or by volume for 5%-50%, 10%-40% or 15%-35% algal biomass or product therefrom.

As described above, microalgae biomass can be replaced by other components, (these components otherwise will be included routinely Include in food article).In some embodiments, food composition contains by weight less than 50%, less than 40% or few In 30% oily or fat, in addition to belonging to biomass or microalgae oil from microalgae source.In some cases, combinations of foods Thing contains by weight less than 25%, less than 20%, less than 15%, the oily or fat less than 10% or less than 5%, except belonging to Biomass or from microalgae source microalgae oil outside.In at least one embodiment, except belonging to biomass or from microalgae Outside the microalgae oil in source, food composition not oil-containing or fat.In some cases, food composition without egg, butter or its His fats/oils or at least one other compositions that would generally be included in comparable conventional food product.Some food articles Without dairy products (for example, butter, cream and/or cheese).

For prepare food composition algal biomass amount depend on to be replaced in conventional food product non-algae oil, Oily percentage in the amount and algal biomass of fat, eggs etc..Therefore, at least one embodiment, side of the invention Method includes generally combining with other at least one comestible compositions according to ratio oily in biomass and in conventional food product Oily and/or fatty ratio determine the amount of algal biomass to be combined with other at least one comestible compositions.For example, such as Fruit algal biomass 50%w/w is microalgae oil, and expects to replace the oily or fat in conventional recipe completely, then can (for example) By 2:1 ratio replaced oil.The ratio can by mass, but for the purpose of practicality, be often easier using measuring cup or Measuring spoon measures volume, and replacement can be by volume.In general, by algal biomass (100/100-X) volume or Quality replaces oily or fatty volume or quality to be replaced, and wherein X is the percentage of microalgae oil in biomass.It is general next Say, oil ＆ fat to be replaced can be replaced all by algal biomass in conventional recipe, but all replace not necessarily, The oily and/or fat of any desired ratio can be retained, remaining part is replaced according to the need for taste and nutrition.Because Algal biomass has contained the protein and phosphatide of emulsifying agent function, it is possible to replaced whole or in part such as with algal biomass The foodstuff of egg etc.If all replacing egg with biomass, occasionally want to or be necessary that with extra emulsifying agent and/or volume Outer water or other liquid improve the emulsification property in food composition, compensating those components that should be provided originally by egg Loss.Because egg is not all fat, for replace egg biomass amount can than for replace it is pure oil or fat The amount of biomass is few.Egg averagely weighs about 58g, includes about 11.2% fat.Therefore, comprising the micro- of the oily about 13g of 50% microalgae Algal biomass can be for whole total fats portions for replacing egg.All or part of eggs are replaced in food article in addition to have There is the benefit of reduction cholesterol.

For simplicity can also be according to the matter of the oil, fat and/or eggs replaced with the quality or volume of biomass Amount or volume provide substitution ratio.In certain methods, with 5%-150%, 25%-100% or 25%-75% quality or body Long-pending oil, fat and/or eggs replace oil in conventional recipe, the quality or volume of fatty and/or eggs.Replacement ratio depends on The whole texture and the oil-containing of outward appearance and biomass of required trophic spectrum, food article in such as food article, food article The factors such as amount.

In ripe food, the determination of percentage (that is, weight or volume) can be carried out before cooking or afterwards.Cooking During preparing food, due to the loss of liquid, the percentage of algal biomass may increase.Because some algal biomass cells can be Dissolved in cooking process, it is possible that it is difficult to directly measuring the content of algal biomass in ripe food.However, the content can be with It is (solid based on biomass as the percentage of finished weight or volume according to the quality or volume for entering the biomass in product Body) and determined indirectly, and by analyzing component specific to algal biomass (such as genome sequence) or by algae biology The method for the compound (such as some carotenoid) that matter is individually delivered is determined.

In some cases, it is desirable to be with more than proportional to oil, fat, eggs etc. present in conventional food product The amount of amount combines algal biomass with other at least one comestible compositions.For example, 1 times, 2 times, 3 times, 4 times or more can be used The amount of times algal biomass replaces the oily and/or fatty quality or volume in conventional food product.Some of the method for the present invention Embodiment is provided for the recipe containing other at least one comestible compositions with the conventional food product of oily or fat composition, And the fatty or oily quality or at least the one of volume by the algal biomass of 1-4 times of quality or volume and in such as conventional food article Plant the combination of other comestible compositions.

By algal biomass (it is mainly complete or homogenize or micronizing) and/or algae oil with it is at least one other can Edible composition combines to form food article.In some food articles, algal biomass and/or algae oil and 1-20,2-10 or 4-8 Other comestible compositions combination.Comestible composition may be selected from all Major Foods classes, including but not limited to water fruits and vegetables, Beans, meat, fish, cereal (for example, wheat, rice, oat, cornflakes, barley), herbaceous plant, spices, water, vegetable soup, fruit Juice, wine and vinegar.In some food compositions, appearance at least 2,3,4 or 5 food qualification category and algal biomass or algae oils.

Oil, fat, eggs etc. can also be incorporated in food composition, but as having been discussed above, with conventional food Tetramune is compared, and they generally exist with the amount of reduction, for example 50%, 25% less than oil, fatty or eggs quality or volume Or 10%.Some food articles of the invention are free of the oil in addition to the oily and/or algae oil provided by algal biomass.Some Food article is free of the oil in addition to the oil provided by algal biomass.Some food articles are free of except by algal biomass or algae Fat beyond the fat that oil is provided.Some food articles are free of the fat in addition to the fat provided by algal biomass.One A little food articles are free of except by both oil ＆ fats in addition to algal biomass or the oil ＆ fat of algae oil offer.Some food systems Product are without both oil ＆ fats in addition to the oil ＆ fat provided by algal biomass.Some food articles are free of eggs. In some embodiments, can be customized by the selection of condition of culture or strain comprising specific fatty acid component or content by The oil that microalgae is produced.

In some cases, at least two different microalgae species are included for making the microalgae biomass of food composition Mixture.In some cases, at least two different microalgae species are separated cultures.In at least one embodiment, At least two different microalgae species have different glyceride spectrums.In some cases, the above method is additionally included in heterotrophism bar Algae is cultivated under part and biomass is prepared by the algae.In some cases, at least two different microalgae species all contain There is by dry weight at least 10% or at least 15% oil.In some cases, food composition contains the biomass of identical type Two different preparations blend, one of preparation contains by dry weight at least 30% oil, and the second preparation contains by dry Oil of the restatement less than 15%.In some cases, food composition contains two different preparations of the biomass of identical type Blend, one of preparation contains by dry weight at least 50% oil, and the second preparation contains oil by dry weight less than 15%, And further, wherein the species is Chlorella protothecoides.

It is same to use algal biomass the same as the oil in other conventional food, fat or eggs sub, algal biomass Available for the oil being typically free of in supplements (such as smoothie).The combination of oil and the product of mainly carbohydrate can have There is the benefit closed with oil phase, and by the combination of oil and carbohydrate, obtained by reducing the glycemic index of carbohydrate Obtain benefit.The measure that oil is wrapped up in biomass is conducive to protection oil not aoxidized, and can also improve the taste of smoothie And texture.

Can be by with using the oil extracted from algal biomass to biomass identical mode itself, that is to say, that as The sub of oil, fat, eggs in conventional recipe etc..The oil can be used for about 1:1 w/w or volume/volume On the basis of replace routine oily and/or fat.The oil can be used for replacing eggs, and mode is that each egg is entered with about 1 algae oil Row is replaced, and optionally with other water and/or emulsifier combination, (average 58g eggs have about 11.2% fat, the density of algae oil to algae oil It is about 0.915g/ml, about 5ml=1.2 algae oil/egg of the volume that teaspoon has).The oil can also be admixed to seasoning Inside product, sauce, soup, margarine, cream, shortening etc..The oil is particularly suitable for use in wherein needing oil and other foods Into subassembly to obtain required taste, texture and/or the food article of outward appearance.By weight or stereometer, oil is in food article Content can be at least 5%, 10%, 25%, 40% or 50%.

In at least one embodiment, the oil extracted from microalgae biomass can also by food production business, restaurant and/ Or consumer is used as edible oil.In this case, algae oil can replace conventional edible oil, such as safflower oil, canola Oil, olive oil, grape-kernel oil, corn oil, sunflower oil, coconut oil, palm oil or any other conventional use of edible oil.Come from The oily and other kinds of oil of algal biomass, it can be subjected to further refining to improve its adaptability (example to culinary art Such as, smoke point is improved).Can be with caustic alkali and oil is to remove free fatty.Free fatty forms removable soap stock. The color for removing oil can be bleached by using chemicals (such as carbon black and bleaching earth).Bleaching earth and chemicals can via filtering with Oil separation.Oily deodorization can also be made by using steam treatment.

Mainly complete biomass, the biomass (as slurry, thin slice, powder or powder) for homogenizing or being micronized and pure The algae oil of change can combine to form food article with other food ingredients.All these is all with favourable trophic spectrum (phase To high single unsaturated content) oil sources.The mainly complete biomass for homogenizing and being micronized additionally provides high-quality Protein (the amino acid composition of balance), carbohydrate, fiber and other nutrients as described above.Can be by absolute element The form of trencherman or vegetarian make the food for being blended with these any products.Using microalgae biomass (it is mainly complete or Homogenize (or micronizing) or have both at the same time) it is that it is vegan/vegetarian diet as another advantage of protein source The protein source of person, is not from main allergen source (such as soybean, eggs or dairy products).

Include but is not limited to paddy according to other comestible compositions that the present invention can be combined with algal biomass and/or algae oil Thing, water fruits and vegetables, protein, meat, herbaceous plant, spices, carbohydrate and fat.With algal biomass and/or algae oil group Close and depend on the food article to be produced and required food article to form other comestible compositions of food composition Taste, texture and other characteristics.

Although in general, these any algae oil sources can be used in arbitrary food article, preferred algae oil source portion Point depend on whether that the oil is mainly used in the mesh of the purpose of nutrition or heat rather than texture, outward appearance or taste for food , or whether intend with the oily taste of food, texture or the outward appearance that combine needed for providing with other food ingredients, and or Instead of improving its nutrition or heat spectrum.

Desired conventional program cooking food product can be passed through.According to cooking time and temperature conditions, cooking process Some cell membranes can be decomposed, are discharged fuel-displaced so that it is combined with the other compositions in mixture.However, often at least some Frustule survives culinary art and keeps complete.Alternatively, food article can be consumed without culinary art.In this feelings Under condition, algae wall keeps complete, and protection oil is not oxidized.

Provided that form to be that cell is mainly complete, or provided as homogenate powder, then microalgae biomass is not It is same as being in place of oil, fat or eggs, it can be provided as dry composition, contribute to the composition (such as powder) with other dryings Mixing.In one embodiment, algal biomass is provided as drying homogenate containing by dry weight 25% to 40% oil.Biomass Homogenate can also be provided as slurry.It can be added after combination drying composition (and biomass juice slurry, if you are using) Liquid (such as water).In some food articles, required amount of liquid is slightly above conventional food article, and this is due to biomass The reason for non-oil ingredient, and/or because water can not be provided by other compositions (such as eggs).However, it is possible to such as in conventional cooking As be readily determined the amount of water.

On the one hand, the present invention relates to (contain by dry weight at least 10% comprising at least 0.5%w/w algal biomass Algae oil) and other at least one comestible compositions food ingredients composition, wherein by the way that liquid is added into food ingredients group Compound can change into food ingredients the food article of recovery.In one embodiment, the liquid is water.

The oily biomass of height for homogenizing or being micronized is in liquid and/or emulsification food article (Water-In-Oil and oil-in-water emulsion) In be it is particularly advantageous, such as sauce, soup, drink, salad-dressing, butter, smear food in, wherein being provided by biomass Oil with other liquid formation emulsion.Benefit from rheological property improved products (such as flavouring, sauce and smearing food) description In the following example., can be relatively low using the biomass homogenized compared with using the situation of the conventional products of conventional oil Being formed under oil content (based on the weight or volume of total product) has required texture (for example, mouthfeel), taste and outward appearance (example Such as, opacity) emulsion, therefore it may be used as fat gain agent.So it is applied to (that is, meals) product low in calories.It is pure The algae oil of change is also advantageous for such liquid and/or emulsifying product.The oily biomass of height that homogenizes or be micronized and Both algae oils of purifying can well be combined with other comestible compositions in baked goods, with conventional oil, fat and/ Or other similar products for being made of eggs are compared to obtaining similar or more preferable taste, outward appearance and texture, but with improved battalion Support spectrum (for example, higher single saturation oil content, and/or higher protein content or quality, and/or higher fiber and/or The content of other nutriments).

Mainly complete biomass be particularly suitable for use in wherein it is desirable to change or improve food trophic spectrum situation (example Such as, higher oil content, different oil contents (for example, more list unsaturated oils), higher protein content, higher heat The content of amount, other higher nutrients).Such food is applicable to (such as) sportsman or with deeline The patient of (wasting disorder).Mainly complete biomass can be used as swelling agent.Swelling agent can be used for (for example) increasing Plus the amount of more expensive food (for example, meat adjuvant etc.), or (the meat generation of such as vegetarian in simulation or imitated food For thing).Simulation or imitated food are different from natural food part and are, local flavor and volume are typically to be provided by different sources 's.For example, the local flavor of natural food (such as meat) can be imparted to inside the swelling agent for keeping local flavor.Mainly complete life Material can be used as swelling agent in this food.Mainly complete biomass is also especially suitable for dried foodstuff, such as Italy Therefore wheaten food, because it has good water binding characteristic, and can promote the rehydration of this food.Mainly complete life Material is also applied for (such as) in baked goods as preservative.Mainly complete biomass can improve water-retaining property, and Therefore extend the shelf life.

Broken or micronizing algal biomass is as a kind of adhesive, swelling agent or change or increases a kind of food article Trophic spectrum can also be useful.Broken algal biomass can be combined with another protein source, such as meat, soybean egg White matter, lactalbumin, wheat gluten, legumin, rice protein, pea protein, lactoprotein etc., wherein the algal biomass conduct A kind of adhesive and/or swelling agent work.The microalgae biomass for having crushed or being micronized can also improve water-retaining property and because This extends the shelf life.Moisture retention is improved to particularly hope in the product (baked goods as be free of seitan) without seitan. The detailed description and follow-up shelf-life research for preparing the cookies without seitan using broken algal biomass are described following In example.

In some cases, algal biomass can be used in egg preparation.In some embodiments, it is added to conventional dry powder egg Have in preparation with the algal biomass (for example, algae powder) for forming creamy scrambled egg compared with the dried egg powder prepared without algal biomass There are more moisture and more preferable texture.In other embodiments, algal biomass is added in egg pulp, to improve The overall texture and humidity of egg that is preparing and being then maintained in steam table.The instantiation of above-mentioned preparation is described in following reality In example.

Can algal biomass (it is mainly complete and/or homogenize or micronizing) and/or algae oil be admixed to reality On border in any food ingredients composition.Some examples include baked goods (such as cake), Brownie, sponge cake, bread (including Cream roll), cookies (including sugared cookies), biscuit and pie.Other examples include the product generally provided with dry form, comply with one's wishes Big profit wheaten food or powdery flavouring, dry milk essence, meat mincing and meat sub.Due to the water combination energy of mainly complete biomass The reason for power, mainly complete biomass being admixed to inside this product can improve as adhesive and/or swelling agent Water, which is combined, improves yield.The rehydrated food (such as scrambled egg) being made up of dried egg powder can also have improved texture and trophic spectrum.Its He includes liquid food products, such as sauce, soup, flavouring (instant), cream, milk drink, fruit drink, smoothie, milk by example Essence.Other liquid food products include serving as the nutritious drink of meals sub or algae milk.Other food articles include butter or Cheese etc., including shortening, margarine/smearing food, nut butter and cheese product (such as Nacho sauce).Other food systems Product include energy stick, chocolate dessert-lecithin sub, meals and replace rod, granola type product.Another food system Category type is pastel and smears food.The oil reservoir of food is surrounded by providing, mainly complete biomass or homogenate is repelled The other oil in medium is cooked, prevents it from penetrating into food.Therefore, food can keep the good of high content list unsaturated oils coating Place is without absorbing undesirable oily (for example, byproduct of trans fats, saturated fat and edible oil).Food is absorbed due to less With oil and its reason for byproduct, biomass be coated can also be provided for food desired texture (for example, crisp) and Pure local flavor.

In raw food, most of frustules in biomass keep complete.This is conducive to protection algae oil not oxidized, Long shelf-life can so be made, and the unfavorable interaction with other compositions is minimized.According to the property of food article, The need for freezing, vacuum packaging etc. can be reduced or avoided in the protective effect assigned by cell.Cell is kept completely can also to prevent oil Directly contacting between consumer's mouth, so reduces undesirable greasy feeling or greasy feel.Make using more oil In the food article of nutritional supplement, can be so conducive to the organoleptic attribute for improving product.Therefore, mainly complete biomass Suitable for this product.However, with oil assign needed for mouthfeel raw food (such as salad-dressing) in (for example, with such as The emulsion of the aqueous solution formation of vinegar etc), the use of the algae oil of purifying or the biomass of micronizing is preferred.In ripe food In, some frustules of originally complete bio matter may dissolve, but other frustules can keep complete.Dissolve cell and complete The ratio of cell depends on temperature and the duration of cooking process.Wishing oil with equal for taste, texture and/or outward appearance In the prepared food thing (for example, baked goods) that even mode is disperseed with other compositions, biomass or the algae of purifying using micronizing Oil is preferred.In the ripe food of oil and/or protein and other nutriments is provided using algal biomass, main mesh Be nutrition or calorific value rather than texture for them.

For the other similar conventional products made without algal biomass, algal biomass is equally applicable to carry The satiety index of high food article (for example, meals replace beverage or smoothie).Satiety index is used to weigh identical calory count Different foods meet the degree of appetite.The metering system of this index can tested food be supplied, hereafter by fixation Time interval measurement to the appetite of other foods.Hereafter the appetite to other foods is smaller, and satiety index is higher.It is full to refer to The expression scale of numerical value can be such, wherein the value of white bread is appointed as into 100.Food with higher satiety index is fitted For going on a diet.Although independent of the understanding to mechanism, algal biomass is considered as the food by improving given heat Protein and/or fiber content and the satiety index for improving food.

Nutritional supplement can also be made in algal biomass (mainly complete and homogenizing or be micronized) and/or algae oil Or edible tonic.For example, can be algae oil parcel into digestible capsule, mode is similar to fish oil.Can be this capsule It is packaged in bottle and day clothes (for example, 1-4 or piece daily).Capsule can the algal biomass containing unit dose or algae oil.Together Sample, optionally can be pressed into tablet by biomass together with medicine or other excipient.The tablet can be packaged in (such as) In bottle or blister package, and by the dosage day clothes of for example daily 1-4 pieces.In some cases, tablet or other formulations are comprising single The biomass or algae oil of position dosage.The production of capsule and tablet and other tonics is preferably being suitable for the GMP of nutritional supplement Under the conditions of carry out, the code being such as organized in 21C.F.R.111 or the similar regulations formulated by foreign jurisdiction.Can be by algae Biomass mixed with other powder and be put into sachet as mixed type material immediately (for example, with water, fruit juice, milk or other Liquid is mixed).Algal biomass can also be mixed into the product of such as yoghourt etc.

Although algal biomass and/or algae oil can be admixed in nutritional supplement, functional food discussed above The typical nutritional supplement of product and ball, capsule or powder type is had any different.No matter the heat side being also to provide in terms of weight For face, the deal of typically this food article is all more much bigger than nutritional supplement.For example, when packaging or one-time-consumption, food Tetramune often has more than 100g weight, and/or provides at least 100 card heats.Typically, food article contains at least one Kind of composition, protein, carbohydrate either liquid, and usually contain two or three it is such other into Point.At least 30%, 50% or 60% of the commonly provided food article heat of protein or carbohydrate in food article.

As discussed above, algal biomass can be made by manufacturer and is sold to consumer (such as restaurant or individual), For commercially receiving in environment or family.This algal biomass is preferably under the conditions of the Good Manufacture Practice (GMP) of food article Production and packaging.Generally in gas-tight container (such as hermetic bag) dry state pack main complete form or homogenized as powder or The algal biomass of Micronised form.In addition to other containers, the biomass for homogenizing or being micronized in slurry form can To be advantageously packaged in pipe.It is optionally possible to pack algal biomass under vacuo to extend the shelf life.Packaged algae life Material need not be freezed.Packaged algal biomass can including the use of specification, including on using how many algal biomass come The oil of specified rate in the conventional recipe of replacement, the explanation of fatty or eggs, as previously described.For the sake of simplicity, specification It can specify that oil or fat by the 2 of biomass quality or volume:1 ratio is replaced, and the ratio that egg is replaced is biological with 11g for each egg Matter or 1 algae oil.As previously described, other ratios are also possible, for example, using the quality or volume pair of biomass The ratio of oily and/or fatty and/or eggs quality or volume in conventional recipe is 10%-175%.Opening is packed Afterwards, specification can instruct user by algal biomass be maintained at gas-tight container (such as can commercially available gas-tight container extensively, for example Glad in), optionally freezed.

Algal biomass can also be packed by the form combined with other dry ingredients (for example, sugar, powder, dry fruit) (mainly Powder that is complete or homogenizing or be micronized) and portion package is carried out, to ensure the uniformity of final products.Then can be by Mixture is transformed into food article by consumer or catering service company, and mode is simply adding liquid (such as water or milk) and to appoint Select and mixed and/or cooked in the case of without addition oil or fat.In some cases, adding liquid is to restore Dry algal biomass composition.Culinary art is optionally using progress on micro-wave oven, convection oven, conventional oven or cooking stove.This Mixture is planted to can be used for making cake, bread, pancake, wafer, beverage, sauce etc..This mixture, which has, conveniently to disappear The person of expense and long shelf-life and the advantage without freezing.This mixture is packaged in the sealing container with specification, For adding liquid so that mixture is transformed into food article.

Equally it is also preferred that producing and packing the algae oil as food ingredients under the conditions of the GMP of food.Generally by algae oil bag In bottle or other containers, mode is similar to routine oil.Container can include the attachment label with specification, specification Have and use oil to replace the conventional oil in food article, fat or eggs and the explanation as edible oil.Hold when being packaged in sealing When in device, oil substantially constant matter with the long shelf-life (at least 1 year).After opening, mainly it is made up of single unsaturated oils Algae oil to oxidation be not very sensitive.If however, keep cold conditions and/or avoid direct sunlight (such as in closing space, As in kas), the oil of unused portion can keep longer time and oxidation is less.The specification included with oil can contain This preferred storage information.

Optionally, preservative/antioxidant that algal biomass and/or algae oil can permit containing food is with to greatest extent Extend the shelf life, the preservative/antioxidant includes but is not limited to carotenoid (for example, astaxanthin, lutein, maize Element, alpha-carotene, beta carotene and lycopene), phosphatide is (for example, N- acylphosphatidyl ethanolamines, phosphatidic acid, phosphatidyl Monoethanolamine, phosphatidyl choline, phosphatidylinositols and lysophosphatidyl choline), tocopherols are (for example, alpha-tocopherol, β-fertility Phenol, Gamma-Tocopherol and Delta-Tocopherol), tocotrienols is (for example, alpha-tocotrienol, β-tocotrienols, γ-fertility triolefin Phenol and δ-tocotrienols), it is Butylated Hydroxytoluene, Butylated Hydroxyanisole, polyphenol, Rosmarinic acid, propylgallate, ascorbic acid, anti-bad Hematic acid sodium, sorbic acid, benzoic acid, methyl p-hydroxybenzoate, levulic acid, anisic acid, acetic acid, citric acid and biological species are yellow Ketone.

In general, by mainly complete biomass, the biomass for homogenizing or being micronized (slurry, thin slice, powder or Powder) or algae oil be admixed to the food article that inhuman animal is can be also suitably used for inside human nutrition food.

Biomass this food is given it is high-quality oil protein or both.The content of algae oil is preferably by weight Meter at least 10% or 20%, as the content of algae albumen.Obtained from mainly complete biomass at least some algae oils and/ Or protein is favourable to the food for high speed animal (such as dog race or horse) sometimes.Mainly complete biomass It is also suitable for preservative.Algal biomass or oil are combined with other compositions and typically see animal food (for example, meat, meat Savory essence, aliphatic acid, veterinary antibiotics, starch, vitamin, mineral matter, antioxidant, probiotic) and their any group Close.This group food is also applied for companion animals, the particularly companion animals with active life style.Recommend to include for cat food Taurine.As conventional animal food, the food in a bite size particles can be provided to appropriate specified animal.

Degreasing meals be as a kind of raw material for being used to producing algae protein concentrate and/or isolate it is useful, especially Degreasing meals from the algal biomass containing high protein.The mark for producing soybean protein concentrate/isolate can be used Quasi- technique produces algae protein concentrate and/or isolate.Made by removing soluble sugar from degreasing algal biomass or meals Standby algae protein concentrate.Remaining ingredient will mainly protein and insoluble polysaccharide.It is soluble by being removed from degreasing meals Sugar, protein content is increased, and thus produces algae protein concentrate.Algae protein concentrate will contain by dry weight at least 45% Protein.Preferably, algae protein concentrate will contain by dry weight at least 50% to 75% protein.Use can also be used Algae Protein Separation thing is prepared in the standard method of production soy protein isolate.This method is usually directed to temperature and used NaOH alkaline pH extraction step.After the extraction step, these liquid and solid are separated and make these eggs using HCl White matter is precipitated out from liquid portion.Solid portion can be extracted again, and can collect what is generated before being precipitated with HCl Liquid portion.Then neutralize the protein and be spray-dried to produce a kind of protein isolate.Algae Protein Separation thing typical case Ground will contain by dry weight at least 90% protein.

Degreasing meals, which are suitable as animal feed, is used for farm-animals, such as ruminant, poultry, pig and aquaculture. Degreasing meals are the byproducts for preparing food or the purifying algae oil of other purposes.Although gained meals oil content is reduced, but still So contain high-quality protein, carbohydrate, fiber, ash content and other nutrients suitable for animal feed.Because cell Mainly dissolve, so this animal is easy to digest degreasing meals., optionally can be by degreasing meals in animal feed Combined with other compositions (such as cereal).Because degreasing meals have powder consistency, it is possible to use extruder or expander will It is pressed into commercially available pill.

Present invention additionally comprises embodiments below：

1. a kind of include the food ingredients composition of dry egg products and algae powder, the algae powder is the even of microalgae biomass Slurry, the microalgae biomass mainly or entirely contain in powder type dissolving cell, including by dry weight at least 16% it is sweet Oily three ester oils, for preparing a kind of food article by adding a kind of liquid and optional other comestible compositions.

2. the food ingredients composition as described in embodiment 1, the egg products of the wherein drying are dry shell eggs.

3. the food ingredients composition as described in embodiment 1, the egg products of the wherein drying are dry albumen.

The 4 food ingredients composition as described in embodiment 1, the egg products of the wherein drying are dry yolk.

5. the food ingredients composition as described in embodiment 1 be a kind of powdered egg products or a kind of pancake or Waffle powder.

6. the food ingredients composition as described in embodiment 1, wherein the algae powder are by the way that microalgae biomass is micronized Form a kind of emulsion and dry formed by the emulsion.

7. the food ingredients composition as described in embodiment 1, wherein the algae powder have the particle mean size less than 100 μm.

8. the food ingredients composition as described in embodiment 7, wherein the particle mean size of the particle in the algae powder is 1- 15μm。

9. the food ingredients composition as described in embodiment 1, the wherein microalgae biomass are 45%-75% by dry weight Triglyceride oil.

10. the food ingredients composition as described in embodiment 1, wherein by weight at least 50% triglyceride oil is 18:1 lipid and in the form of glyceride and by comprising.

11. the food ingredients composition as described in embodiment 1, wherein the triglyceride oil by weight less than 5% is Docosahexaenoic acid (DHA) (22:6).

12. the food ingredients composition as described in embodiment 1, the wherein triglyceride oil are：

A. 2%14 are less than:0；

B.13% to 16%16:0；

C.1% to 4%18:0；

D.64% to 70%18:1；

E.10% to 16%18:2；

F.0.5% to 2.5%18:3；And

G. the carbon chain lengths less than 2% are 20 or longer oil.

13. the food ingredients composition as described in embodiment 1, the wherein microalgae biomass be by dry weight 25% to Carbohydrate between 40%.

14. the food ingredients composition as described in embodiment 1, the wherein microalgae biomass include a kind of carbohydrate Component, the carbohydrate ingredient contains by dry weight between the dietary fiber and 2% to 8% between 25% to 35% Free sugar including sucrose.

15. the food ingredients composition as described in embodiment 1, the wherein microalgae biomass include dietary fiber, the meals There is food fiber following monose to constitute：

A.0.1% to 3% arabinose；

B.5% to 15% mannose；

C.15% to 35% galactolipin；And

D.50% to 70% glucose.

16. the food ingredients composition as described in embodiment 1, the wherein microalgae biomass have in 20 to 115 μ g/g Between total carotinoid, include 20-70 μ g/g lutein.

17. the leaf of the food ingredients composition as described in embodiment 1, the wherein microalgae biomass having less than 10 μ g/g Flavine.

18. the food ingredients composition as described in embodiment 1, the wherein microalgae biomass are included less than 2ppm contents Chlorophyll.

19. the food ingredients composition as described in embodiment 1, the wherein microalgae biomass have 1-8mg/100g's total Tocopherols, include 2-6mg/100g alpha tocopherol.

20. the food ingredients composition as described in embodiment 1, the wherein microalgae biomass have 0.05-0.30mg/g Total tocotrienols, include 0.10-0.25mg/g α tocotrienols.

21. the food ingredients composition as described in embodiment 1, the wherein microalgae biomass are derived from the kind of Chlorella The microalgae of class.

22. the food ingredients composition according to embodiment 21, the wherein microalgae are a strains of Chlorella protothecoides.

23. the food ingredients composition as described in embodiment 1, the wherein microalgae biomass are derived from and from derived from it A kind of algae with the reduced colour mutant of pigment formation is compared in strain.

24. the food ingredients composition as described according to embodiment 23, the wherein algae do not have visible green or yellow.

25. one kind is by combining food ingredients composition formed by egg products and algae powder, the algae powder is microalgae biomass Homogenate, the microalgae biomass mainly or entirely contains the dissolving cell in powder type, including by dry weight at least 16% Triglyceride oil, for preparing a kind of food system by adding a kind of liquid and optional other comestible compositions Product.

26. the food ingredients composition as described in embodiment 25 is a kind of pasta.

27. a kind of food ingredients composition including liquid egg products and algae slurry material, wherein the algae powder are that microalgae is biological The homogenate of matter, the microalgae biomass mainly or entirely contains the dissolving cell in powder type, including by dry weight at least 16% triglyceride oil.

28. the food ingredients composition as described in embodiment 27, wherein the liquid egg products are liquid shell egg, liquid egg In vain, liquid yolk or liquid egg substitute.

29. the food ingredients composition as described in embodiment 27, the preparation for the scrambled egg product in heating.

30. a kind of method for preparing food article, including include the food ingredients and microalgae powder of drying egg products by a kind of Combine and cook with a kind of liquid and optional other comestible compositions, the microalgae powder is the homogenate of microalgae biomass, Mainly or entirely contain the dissolving cell in powder type, including at least 16% triglyceride oil by dry weight.

31. the method as described in embodiment 30, the wherein food article are powdered egg products or a kind of pancake Or waffle powder.

32. a kind of slurry by combining egg products and microalgae powder or microalgae powder and at least one other edibles into Point and food composition formed by heating, the wherein microalgae powder is the homogenate of microalgae biomass, and the microalgae biomass is main Or contain the dissolving cell in powder type, including at least 16% triglyceride oil by dry weight completely.

33. the food composition as described in embodiment 32, the wherein egg products are a kind of liquid egg products.

34. the food composition as described in embodiment 33, wherein the liquid egg products be liquid shell egg, liquid yolk, Liquid protein or liquid egg substitute.

35. the food composition as described in embodiment 32, the wherein egg products are a kind of dry egg products.

36. the food composition according to embodiment 35, wherein the drying egg products are dry shell egg, drying Yolk or the albumen of drying.

37. a kind of food ingredients composition including egg products and algae powder, the algae powder is the homogenate of microalgae biomass, and this is micro- Algal biomass mainly or entirely contains the dissolving cell in powder type, including is no more than 20% triglycerides by dry weight Oil and by dry weight at least 40% protein, for by adding a kind of edible liquid and optional other edibles Composition prepares a kind of food article.

38. the food article of seitan is free of formed by a kind of combination by microalgae biomass, including by dry weight extremely Few 16% triglyceride oil and at least one other powder without seitan or the cereal products without seitan.

39. the food article as described in embodiment 38, wherein should the powder without seitan or the cereal products without seitan Including at least one：Three-coloured amaranth powder, kudzuvine root starch, buckwheat, ground rice, olecranon bean powder, cornflakes, cornflour, millet powder, Ma Ling Potato powder, farina, quinoa powder, sorghum flour, soy meal, bean powder, legume flour, cassava (cassava) powder, Ethiopia Russia Than sub- thrush grass meal, globe artichoke powder, almond powder, acorn flour, coconut powder, chest nut powder, corn flour or taro meal.

40. the food article as described in embodiment 38, the wherein microalgae biomass are a kind of microalgae powders.

41. the food article as described in embodiment 40, the wherein microalgae powder have the average grain between 1 and 100 μm Degree.

42. the food article as described in embodiment 38, the wherein triglyceride oil are：

A. 2%14 are less than:0；

B.13% to 16%16:0；

C.1% to 4%18:0；

D.64% to 70%18:1；

E.10% to 16%18:2；

F.0.5% to 2.5%18:3；And

G. the carbon chain lengths less than 2% are 20 or longer oil.

43. the food article as described in embodiment 38, the wherein microalgae biomass have by dry weight 25% to Carbohydrate between 40%.

44. the food article as described in embodiment 38, the wherein microalgae biomass include dietary fiber, the dietary fiber Following monose is constituted：

A.0.1% to 4% arabinose；

B.5% to 15% mannose；

C.15% to 35% galactolipin；And

D.50% to 70% glucose.

45. the food article as described in embodiment 38, the wherein microalgae biomass have between 20 to 115 μ g/g Total carotinoid, includes 20-70 μ g/g lutein.

46. the lutein of the food article as described in embodiment 38, the wherein microalgae biomass having less than 10 μ g/g.

47. the food article as described in embodiment 38, the wherein microalgae biomass include green less than the leaf of 2ppm contents Element.

48. the food article as described in embodiment 38, the wherein microalgae biomass have 1-8mg/100g total fertility Phenols, includes 2-6mg/100g alpha tocopherol.

49. the food article as described in embodiment 38, the wherein microalgae biomass have 0.05-0.30mg/100g's Total tocotrienols, includes 0.10-0.25mg/g α tocotrienols.

50. the food article according to embodiment 40, the wherein microalgae powder do not have visible oil.

51. the food article as described in embodiment 38, the wherein microalgae biomass are derived from the species of Chlorella A kind of microalgae.

52. the food article according to embodiment 51, the wherein microalgae are Chlorella protothecoides.

53. the food article as described in embodiment 38, the wherein microalgae biomass are derived from a kind of microalgae, with spreading out from it Raw strain is with the reduced colour mutant of pigment formation compared to the microalgae.

54. the food article as described in embodiment 38, the wherein triglyceride oil are less than 5% two dodecahexaenes Acid (DHA) (22:6).

55. the food article as described in embodiment 38, the wherein microalgae biomass are mainly dissolving cell.

56. the food article as described in embodiment 38, be a kind of baked goods, bread, cereal preparation, crispbread or Pasta.

57. the food article as described in embodiment 56, the wherein food article are a kind of baked goods being selected from the group, The group is made up of the following：Brownie, cake and pie product and cookies.

58. the food article as described in embodiment 38, without the oil in addition to the algae oil for belonging to the microalgae biomass Or fat.

59. the food article as described in embodiment 38, without yolk.

60. the food article as described in embodiment 38, is a kind of product of life.

61. the food article as described in embodiment 38, is a kind of ripe product.

62. a kind of compound powder without seitan, including a kind of microalgae powder and at least one other in addition to microalgae powder The powder without seitan, the wherein microalgae powder includes the homogenate of microalgae biomass, and the microalgae biomass is mainly or entirely containing place Dissolving cell in powder type and the triglyceride oil containing by dry weight at least 16%.

63. the compound powder without seitan as described in embodiment 62, wherein at least one others are free of seitan Powder is to be selected from the group, and the group is made up of the following：Three-coloured amaranth powder, kudzuvine root starch, buckwheat, ground rice, olecranon bean powder, cornflakes, jade Chinese sorghum powder, millet powder, dehydrated potato powder, farina, quinoa powder, sorghum flour, soy meal, bean powder, legume flour, wood Potato (cassava) powder, eragrosits abyssinica powder, globe artichoke powder, almond powder, acorn flour, coconut powder, chest nut powder, corn flour and Taro meal.

64. the compound powder without seitan as described in embodiment 62, the wherein microalgae powder have 1 and 100 μm it Between particle mean size.

65. the compound powder without seitan as described in embodiment 62, the wherein microalgae powder have by weight 10% Or less or 5% or less moisture.

66. the compound powder without seitan as described in embodiment 62, the wherein microalgae biomass have by dry weight Triglyceride oil between 45% to 70%.

67. the compound powder without seitan as described in embodiment 62, wherein 60% to 75% triglyceride oil is 18 in glyceride form:1 lipid.

68. the compound powder without seitan as described in embodiment 62, the wherein triglyceride oil are：

A. 2%14 are less than:0；

B.13% to 16%16:0；

C.1% to 4%18:0；

D.64% to 70%18:1；

E.10% to 16%18:2；

F.0.5% to 2.5%18:3；And

G. the carbon chain lengths less than 2% are 20 or longer oil.

69. the compound powder without seitan as described in embodiment 62, the wherein microalgae biomass include being less than 2ppm The chlorophyll of content.

70. the compound powder without seitan as described in embodiment 62, wherein said composition do not have visible oil.

71. the compound powder without seitan as described in embodiment 62, further comprises glidant.

72. the compound powder without seitan as described in embodiment 62, the wherein microalgae biomass are derived from microalgae Single strain no more than one.

73. the compound powder without seitan as described in embodiment 62, the wherein microalgae biomass are derived from chlorella The algae of the species of category.

74. the compound powder without seitan according to embodiment 73, the wherein algae are Chlorella protothecoides.

75. the compound powder without seitan as described in embodiment 62, the wherein microalgae biomass be derived from from its A kind of algae with the reduced colour mutant of pigment formation is compared in derivative strain.

76. a kind of method that mitigation seitan does not tolerate symptom, including：

A. with by combining the same-type that microalgae biomass is produced in the diet for the subject not tolerated with seitan Food article substitution the food article containing seitan, the microalgae biomass include on dry basis at least 16% triglycerides Oily and at least one other food ingredients for being free of seitan, wherein same type of food article is free from seitan；And And

B. the same type of food article is provided to the subject that seitan is not tolerated；

Thus the symptom that at least one seitan in the subject is not tolerated is reduced.

77. it is a kind of by bakeing microalgae powder and a kind of edible liquid and at least one other comestible composition Food article formed by mixture, the microalgae powder is the homogenate of microalgae biomass, and the microalgae biomass mainly or entirely contains Dissolving cell in powder type, with by weight at least 16% triglycerides oil content, mainly or entirely containing molten Solve cell.

78. the food article as described in embodiment 77, it has the water activity (Aw) between 0.3 and 0.95.

79. the food article as described in embodiment 77, compared with a kind of other identical conventional food product, it has There are at least 1.5 times higher fiber contents.

80. the food article as described in embodiment 77, is to be selected from the group, the group is made up of the following：Brownie, song Very, cake and pie product, crispbread, bread and snack chip.

81. the food article as described in embodiment 80, wherein bread are pizza crust, bread stick, cream roll or cake It is dry.

82. the food article as described in embodiment 77, the wherein microalgae biomass are the sweet of 45%-75% by dry weight Oily three ester oils.

83. the food article as described in embodiment 82, wherein by weight at least 50% triglyceride oil is a kind of 18:1 lipid and in the form of glyceride and by comprising.

84. the food article as described in embodiment 77, wherein the algae oil less than 5% is docosahexaenoic acid (DHA) (22:6)。

85. the food article as described in embodiment 77, wherein 60% to 75% triglyceride oil is to be in glyceride The 18 of form:1 lipid.

86. the food article as described in embodiment 77, the wherein triglyceride oil are：

A. 2%14 are less than:0；

B.13% to 16%16:0；

C.1% to 4%18:0；

D.64% to 70%18:1；

E.10% to 16%18:2；

F.0.5% to 2.5%18:3；And

G. the carbon chain lengths less than 2% are 20 or longer oil.

87. according to the food article described in embodiment 77, the wherein microalgae biomass is by dry weight 25% to 40% Between carbohydrate.

88. the food article as described in embodiment 77, the wherein microalgae biomass have between 20 to 115 μ g/g Total carotinoid, includes 20-70 μ g/g lutein.

89. the lutein of the food article as described in embodiment 77, the wherein microalgae biomass having less than 10 μ g/g.

90. the food article as described in embodiment 77, the wherein microalgae biomass include green less than the leaf of 2ppm contents Element.

91. the food article according to embodiment 90, the wherein algae do not have visible green or yellow.

92. the food article as described in embodiment 77, the wherein microalgae biomass have 1-8mg/100g total fertility Phenols, includes 2-6mg/100g alpha tocopherol.

93. the food article as described in embodiment 77, the wherein microalgae biomass have 0.05-0.30mg/g total life Triolefin phenols is educated, includes 0.10-0.25mg/g α tocotrienols.

94. the food article as described in embodiment 77, the wherein microalgae biomass are derived from the species of Chlorella Microalgae.

95. the food article as described in embodiment 94, the wherein microalgae are a strains of Chlorella protothecoides.

96. the food article as described in embodiment 77, the wherein microalgae biomass be derived from from its derivative strain phase Than a kind of algae with the reduced colour mutant of pigment formation.

97. the food article as described in embodiment 77, further a kind of to include selected from chemical leavening agent or biological swollen hair The leavening agent of agent.

98. the food article as described in embodiment 77, the wherein microalgae biomass include at least 40% protein.

99. a kind of method for manufacturing baking goodses, including:

Including microalgae biomass is combined with a kind of edible liquid and at least one other comestible compositions, the microalgae Biomass has by weight at least 25% triglycerides oil content, in full cell sheet or full cell powder or in powder The micronizing homogenate of last form, and

Bakee the mixture.

100. the method as described in embodiment 99, the wherein baking goodses are Brownie, cookies, cake, bread, Piza Cake skin, bread stick, crispbread, biscuit, pie crust or snack chip.

101. the method as described in embodiment 99, the wherein microalgae biomass not with a kind of edible liquid or other Comestible composition combination, other comestible compositions be mainly fat, oil or egg.

102. a kind of beverage including microalgae biomass and edible liquid, the microalgae biomass contains by dry weight at least 16% triglyceride oil and/or at least 40% in full cell or mainly or entirely containing dissolving cell homogenates form Dried protein.

103. the beverage as described in embodiment 102, the wherein microalgae biomass are the forms of the homogenate in micronizing.

104. the homogenate of the beverage as described in embodiment 103, the wherein micronizing is with the average grain less than 100 μm Degree.

105. the beverage as described in embodiment 104, the wherein particle mean size are 1-15 μm.

106. the beverage as described in embodiment 102, the wherein microalgae biomass are derived from the microalgae of heterotrophic growth.

107. the beverage as described in embodiment 102, the wherein biomass are by mass spectral analysis without the algae poison that can be detected Element.

108. the beverage as described in embodiment 102, is pasteurizing.

109. the beverage as described in embodiment 102, further comprises a kind of exogenous proteins source and/or lactose.

110. the beverage as described in embodiment 109, wherein the exogenous proteins source is lactalbumin.

111. the beverage as described in embodiment 102, the wherein beverage are free of lactose.

112. the beverage as described in embodiment 102, the wherein beverage are to be selected from the group, the group is made up of the following： Milk, fruit juice, smoothie, nutritious drink, eggnog and meal replacement beverage.

113. the beverage as described in embodiment 102, the wherein microalgae biomass be by dry weight 45% to 75% it is sweet Oily three ester oils.

114. the beverage as described in embodiment 102, wherein by weight at least 50% triglyceride oil is one kind 18: 1 lipid and in the form of glyceride and by comprising.

115. the beverage as described in embodiment 102, wherein 60% to 75% triglyceride oil, which is one kind, is in glycerine The 18 of ester-formin:1 lipid.

116. the beverage as described in embodiment 102, the wherein triglyceride oil are：

A. 2%14 are less than:0；

B.13% to 16%16:0；

C.1% to 4%18:0；

D.64% to 70%18:1；

E.10% to 16%18:2；

F.0.5% to 2.5%18:3；And

G. the carbon chain lengths less than 2% are 20 or longer oil.

117. the beverage as described in embodiment 102, the wherein microalgae biomass be by dry weight 25% to 40% it Between carbohydrate.

118. the beverage as described in embodiment 102, the wherein microalgae biomass have total between 20 to 115 μ g/g Carotenoids, includes 20-70 μ g/g lutein.

119. the lutein of the beverage as described in embodiment 102, the wherein microalgae biomass having less than 10 μ g/g.

120. the beverage as described in embodiment 102, the wherein microalgae biomass include the chlorophyll less than 2ppm contents.

121. the beverage as described in embodiment 102, the wherein microalgae biomass are derived from a single strain of microalgae.

122. the beverage as described in embodiment 121, the wherein microalgae are a strains of Chlorella protothecoides.

123. the beverage as described in embodiment 102, the wherein microalgae biomass are derived from compared with from its derivative strain A kind of algae with the reduced colour mutant of pigment formation.

124. the beverage as described in embodiment 123, the wherein algae do not have visible green or yellow.

125. the beverage as described in embodiment 102, the wherein edible liquid are soya-bean milk, Rice ＆ peanut milk or almond milk.

126. a kind of method for manufacturing beverage, including in full cell sheet or powder type microalgae biomass or will have The homogenate of the micronizing in powder type of at least 25% triglycerides oil content combines to form one with a kind of edible liquid Plant beverage.

127. a kind of food article of fermentation, including：

A. containing at least 16% triglyceride oil by dry weight and/or by dry weight at least 40% in full cell Or the microalgae biomass of the protein of the form of the homogenate mainly or entirely containing dissolving cell；

B. a kind of edible liquid；And

C. it is a kind of suitable for the microorganism of the work used in food article.

128. the food article of the fermentation as described in embodiment 127, it is Yoghourt.

129. the food article of the fermentation as described in embodiment 128, the wherein Yoghourt are the forms in liquid beverage.

130. a kind of microalgae powder, it is the microalgae biomass for mainly or entirely containing the dissolving cell in powder type Homogenate, the wherein microalgae biomass include at least 40% protein by dry weight and are less than 20% glycerine three by dry weight Ester oil, and wherein the microalgae biomass be derived under the conditions of Good Manufacture Practice (GMP) Heterotrophic culture and processing Algae, the wherein microalgae biomass are derived from the one of the colour mutant for having pigment formation reduced compared with from its derivative strain Plant algae.

131. the microalgae powder as described in embodiment 130, the wherein powder have 1-15 μm of particle mean size.

132. the microalgae powder as described in embodiment 130, the wherein microalgae biomass include by weight at least 10% Dietary fiber.

133. the microalgae powder as described in embodiment 130, the wherein protein are at least 40% digestible crude protein matter.

134. the microalgae powder as described in embodiment 130, the wherein microalgae biomass are derived from Chlorella protothecoides.

135. the microalgae powder as described in embodiment 130, the wherein microalgae biomass be derived from microalgae be no more than one Single strain.

136. the microalgae powder as described in embodiment 130, the wherein microalgae biomass include green less than the leaf of 2ppm contents Element.

137. the microalgae powder as described in embodiment 136, the wherein algae do not have visible green or yellow.

138. a kind of food composition, by the way that other are edible by the microalgae powder as described in embodiment 130 and at least one Formed with into subassembly.

139. the food composition as described in embodiment 138, is meat analogue product, protein rod or the nutrition drink of vegetarian Material.

140. a kind of method for manufacturing vegetarian's meat analogue product, this method includes：By including by dry weight at least 40% The microalgae biomass of protein and triglyceride oil less than by dry weight 20% and at least one other vegetarian's protein sources Combination, also, wherein the microalgae biomass be derived under the conditions of Good Manufacture Practice (GMP) Heterotrophic culture and processing Microalgae.

A kind of 141. foods or food ingredients composition, it contains the even of by weight at least 10% microalgae biomass Slurry, the microalgae biomass is mainly or entirely containing dissolving cell, including at least 16% breast in edible liquid by dry weight The triglyceride oil of change.

142. composition as described in embodiment 141, it is sauce, mayonnaise, soup or flavouring.

143. composition as described in embodiment 141, it is free of the oil in addition to the oil in the microalgae biomass And fat.

144. composition as described in embodiment 141, containing except belong to the microalgae biomass oil in addition to by weight Gauge is less than 25% oily or fat.

145. composition as described in embodiment 144, containing except belong to the microalgae biomass oil in addition to by weight Gauge is less than 10% oily or fat.

146. composition as described in embodiment 141, it is oil-in-water emulsion.

147. composition as described in embodiment 141, it is water-in-oil emulsion.

148. composition as described in embodiment 141, the wherein microalgae biomass are by dry weight 45% to 75% Triglyceride oil.

149. composition as described in embodiment 141, wherein by weight at least 50% triglyceride oil is a kind of 18:1 lipid and in the form of glyceride and by comprising.

150. composition as described in embodiment 149, wherein 60% to 75% triglyceride oil is a kind of in sweet The 18 of grease form:1 lipid.

151. composition as described in embodiment 141, the wherein triglyceride oil are：

A. 2%14 are less than:0；

B.13% to 16%16:0；

C.1% to 4%18:0；

D.64% to 70%18:1；

E.10% to 16%18:2；

F.0.5% to 2.5%18:3；And

G. the carbon chain lengths less than 2% are 20 or longer oil.

152. be by dry weight 25% to 40% according to the composition described in embodiment 141, the wherein microalgae biomass Between carbohydrate.

153. composition as described in embodiment 141, the wherein microalgae biomass include green less than the leaf of 2ppm contents Element.

154. composition as described in embodiment 153, the wherein microalgae biomass do not have visible green or yellow.

155. composition as described in embodiment 141, the wherein microalgae biomass are derived from the microalgae of Chlorella.

156. composition as described in embodiment 155, the wherein microalgae are a strains of Chlorella protothecoides.

157. composition as described in embodiment 156, the wherein microalgae are to preserve title on October 13rd, 2009 The Chlorella protothecoides 33-55 of American type culture collection is deposited under PTA-10397.

158. composition as described in embodiment 156, the wherein microalgae are to preserve title on October 13rd, 2009 The Chlorella protothecoides 25-32 of American type culture collection is deposited under PTA-10396.

159. by slurry formed by scattered algae powder, and it is the homogenate of microalgae biomass, the microalgae biomass it is main or Completely containing dissolving cell, including at least 16% triglyceride oil for being in powder type in aqueous by dry weight, its In the algae powder constitute by weight the 10% to 50% of the slurry.

160. slurry as described in embodiment 159, the wherein microalgae biomass have 5% to 55% glycerine by dry weight The oil content of three ester oils.

161. slurry as described in embodiment 159, the wherein microalgae biomass include by weight at least 40% egg White matter, and the protein includes at least 60% digestible crude protein matter.

The method that a kind of 162. manufactures include microalgae biomass food article, including：

Based on the amount of oil, fat or egg in the food article of conventionally form, it is determined that will be included in the food article Microalgae biomass amount, wherein the microalgae biomass comprising by dry weight at least 16% triglyceride oil；And

The microalgae biomass of the amount combine with one or more comestible compositions (and be less than and be present in conventionally form The amount of oil in food article, fat or egg) to form the food article that this includes microalgae biomass.

A kind of 163. microalgae triglyceride oils for the purifying for being suitable for human consumption, it include at least 50% oleic oil with And the DHA less than 5%, wherein microalgae oil is without the microalgae toxin that can be detected and is prepared under good manufacturing condition 's.

The 164. microalgae triglyceride oil as described in embodiment 163, the wherein oil is：

A. 2%14 are less than:0；

B.13% to 16%16:0；

C.1% to 4%18:0；

D.64% to 70%18:1；

E.10% to 16%18:2；

F.0.5% to 2.5%18:3；And

G. the carbon chain lengths less than 2% are 20 or longer oil.

The 165. microalgae triglyceride oil as described in embodiment 163, it is derived from Chlorella protothecoides.

The 166. microalgae triglyceride oil as described in embodiment 163, further comprises a kind of antioxidant of addition.

The 167. microalgae triglyceride oil as described in embodiment 163, the wherein oil have between 40 to 230 μ g/g Total carotinoid, includes 40-70 μ g/g lutein.

The 168. microalgae triglyceride oil as described in embodiment 163, wherein chlorophyll of the oil having less than 2ppm.

The 169. microalgae triglyceride oil as described in embodiment 163, the wherein oil have 2 between 16mg/100g Total tocopherols, include 4-12mg/100g alpha tocopherol.

The 170. microalgae triglyceride oil as described in embodiment 163, the wherein oil have 0.10 between 0.6mg/g Total tocotrienols, include 0.2-0.5mg/g α tocotrienols.

A kind of 171. smearing foods, including microalgae triglyceride oil and a kind of liquid as described in embodiment 163, wherein The oil and the liquid are formed a kind of emulsion of stabilization.

The 172. smearing food as described in embodiment 171, further comprises a kind of emulsifying agent.

The 173. smearing food as described in embodiment 171, wherein the smearing food can be smeared at ambient temperature.

The 174. smearing food as described in embodiment 171, wherein the smearing food are to apply at a temperature of 5 DEG C -10 DEG C Smear.

175. a kind of margarines, by the microalgae triglyceride oil for the purifying for making to produce under the conditions of Good Manufacture Practice A kind of chemical reaction or enzymatic reaction are subjected to, the margarine is thus produced.

176. margarine as described in embodiment 175, the wherein chemical reaction are hydrogenizations.

177. margarine as described in embodiment 175, the wherein chemical reaction or enzymatic reaction be with from microalgae The glyceride of the different lipodogrammes of triglyceride oil carries out ester exchange.

178. margarine as described in embodiment 177, wherein the different lipodogrammes from microalgae triglyceride oil Glyceride comes from the one or more oil being selected from the group, and the group is made up of the following：Soybean, rapeseed, low erucic acid dish Seed, palm, palm kernel, coconut, corn, olive, sunflower, cottonseed, calyx away from flower, peanut, False flax, mustard seeds, cashew nut, oat, Lupin, mestha, pot marigold, hemp, coffee, linseed, fibert, the root of Beijing euphorbia, pumpkin, coriander, camellia, sesame, safflower, water Rice, tung tree, cocoa, copra, opium poppy, castor bean, hickory nut, jojoba, manioca, Queensland nut, Bertholletia excelsa And avocado.

A kind of 179. animal food products, it by by degreasing microalgae biomass and one or more other edibles into Subassembly and formed, wherein the degreasing microalgae biomass constitute all the components of the animal food product by dry weight at least 0.1%.

The 180. animal food product as described in embodiment 179, the wherein one or more others comestible composition Including cereal.

The 181. animal food product as described in embodiment 179, it is prepared for farm-animals.

Following instance is provided to illustrate but do not limit claimed invention.

V. embodiment

Example 1

Microalgae is cultivated to obtain floorboard with high oil content

Microalgae strain is cultivated in shaking flask, target is to obtain the oil by dry cell weight basis more than 20%.In shaking flask used Culture medium is as follows: K₂HPO₄:4.2g/L, NaH₂PO₄:3.1g/L, MgSO₄·7H₂O:0.24g/L, monohydrate potassium:0.25g/ L, CaCl₂2H₂O:0.025g/L, yeast extract:2g/L and 2% glucose.Freeze-stored cell is set to thaw at room temperature, to 500ul cells are added in 4.5ml culture mediums, are grown 7 days in 28 DEG C on 6 orifice plates, while stirring (200rpm).By advance Centrifuge 1ml culture 5 minutes to determine dry cell weight with 14,000rpm in the Eppendorf tube weighed.Discard culture supernatant Liquid, gained cell precipitation 1ml deionized water rinsings.Culture is centrifuged again, and cell precipitation is placed in -80 by abandoning supernatant DEG C until freeze.Then freeze drying example 24 hours and dry cell weight is calculated.To determine the total lipid in culture, take out 3ml cultures are simultaneously analyzed using Ankom systems (Ankom Inc., Macedon, NY) according to the scheme of producer.Use Amkom XT10 extractors carry out solvent extraction by producer's scheme to sample.Total lipid is defined as acid-hydrolyzed drying sample with being carried through solvent It is of poor quality between the drying sample taken.It is shown in Table 1 by the measurement result of the oily percentage of dry cell weight basis.

1. table presses the oily percentage of dry cell weight basis

Also the other strain for making Chlorella protothecoides using above-mentioned condition grows, and uses what is briefly described in example 2 The lipodogramme of standard gas chromatography (GC/FID) step measurements these Chlorella protothecoides strain.The general introduction of lipodogramme includes as follows. Value is expressed as the area percentage of total lipid.There is UTEX preserving number to come comfortable University of Texas at Austin for these The algae of the UTEX algae collections of (1University Station A6700, Austin, Texas 78712-0183) Strain.There is CCAP preserving number to be from algae and the algae of protozoan culture collection strain (SAMS research services for these Scotland institute of oceanography of Co., Ltd, OBAN, Argull PA371QA, Scotland, Britain).These have SAG preserving number It is the algal cultures collection (the brother's Dettingens of Nikolausberger Weg 18,37073, Germany) from brother's Dettingen university Algae strain.

Although these data show that all strains of the above are all Chlorella protothecoides, the lipodogramme between some plants is present Difference.

Example 2

In order to generate the algal biomass with floorboard with high oil content, three are carried out with three kinds of different culture medium prescriptions and had fermented Journey.First pharmaceutical formulation (culture medium 1) is based on culture medium (the 1994Science in China, volume 37, described in Wu etc. 3 phases, the 326-335 pages), every liter of composition is：KH₂PO₄, 0.7g；K₂HPO₄, 0.3g；MgSO₄-7H₂O, 0.3g；FeSO₄- 7H₂O,3mg；Thiamine hydrochloride, 10 μ g；Glucose, 20g；Glycine, 0.1g；H₃BO₃, 2.9mg；MnCl₂-4H₂O, 1.8mg； ZnSO₄-7H₂O, 220 μ g；CuSO₄-5H₂O, 80 μ g；And NaMoO₄-2H₂O, 22.9mg.Second culture medium (culture medium 2) is source The culture medium in shaking flask in example 1, every liter of composition is：K₂HPO₄, 4.2g；NaH₂PO₄, 3.1g；MgSO₄-7H₂O, 0.24g；Monohydrate potassium, 0.25g；Chloride dehydrate calcium, 25mg；Glucose, 20g；Yeast extract, 2g.3rd culture medium (culture medium 3) is mixed culture medium, and every liter of composition is：K₂HPO₄, 4.2g；NaH₂PO₄,3.1g；MgSO₄-7H₂O, 0.24g；One Citric acid monohydrate, 0.25g；Chloride dehydrate calcium, 25mg；Glucose, 20g；Yeast extract, 2g；H₃BO₃, 2.9mg；MnCl₂- 4H₂O, 1.8mg；ZnSO₄-7H₂O, 220 μ g；CuSO₄-5H₂O, 80 μ g；And NaMoO₄-2H₂O, 22.9mg.Prepare it is all this three Kind of culture medium and in the fermentor vessel of laboratory scale in 121 DEG C of autoclavings 30 minutes.Autoclaving each holds backward Sterile dextrose is added in device, is then cooled down.

Inoculum for each fermentation tank is Chlorella protothecoides (UTEX 250), and its preparation is entered by two flask stages OK, using inoculation fermentation tank culture medium and temperature conditionss.Each fermentation tank is entered with 10% (v/v) mid-log phase culture Row inoculation.The fermentation tank of three laboratory scales is maintained at 28 DEG C in duration of experiment.Also assessed at a temperature of 23 DEG C Microalgae cell growth in culture medium 1.Assessed for all fermentation tanks, pH value is maintained at 6.6-6.8, stirred with 500rpm, Air mass flow is 1vvm.Culture to fermentation culture medium progress 11 days.Measure biological by 750nm optical density and dry cell weight Matter is accumulated.

By the gas chromatography of standard, lipid/oil concentration is determined using direct ester exchange.Briefly, with life The fermentation broth sample of material is absorbed on blotting paper, is transferred in centrifuge tube and that 1 is dried in 65 DEG C of -70 DEG C of vacuum drying ovens is small When.When drying sample, 5%H in methyl alcohol is added into pipe₂SO₄2ml.Then institute is heated on 65 DEG C of -70 DEG C of heat blocks Pipe is stated, while being intermittently vortexed and sonication.Then add 2ml heptane and firmly shake pipe.Add 2Ml 6%K₂CO₃ And firmly shake pipe is mixed, and is then centrifuged 2 minutes with 800rpm.Then supernatant is transferred to containing Na₂SO₄Drier In GC bottles, tested using the gas chromatography of standard.Oil/lipid percentage is to be based on dry cell weight.For growth Cell the following dry cell weight of use：Culture medium 1 is 9.4g/L at 23 DEG C；Culture medium 1 is 1.0g/L, culture at 28 DEG C Base 2 is 21.2g/L at 28 DEG C；And culture medium 3 is 21.5g/L at 28 DEG C.For the following fat of use of the cell of growth Matter/oil concentration：Culture medium 1 is 3g/L at 23 DEG C；Culture medium 1 is 0.4g/L at 28 DEG C；Culture medium 2 is 18g/ at 28 DEG C L；Culture medium 3 is 19g/L at 28 DEG C.The oily percentage below based on dry cell weight is used for the cell of growth：Culture medium 1 It is 32% at 23 DEG C；Culture medium 1 is 40% at 28 DEG C, and culture medium 2 is 85% at 28 DEG C；And culture medium 3 is at 28 DEG C It is 88% down.Using three kinds of different culture medium prescriptions 28 DEG C generation algal biomass lipodogramme (area %, internal standard normalizing After change) it is summarized in table 2 below.

The lipodogramme for the Chlorella protothecoides that table 2. grows under the conditions of different culture media

Example 3

Prepare the biomass for food article

As described in any one of example 1-2, microalgae biomass is produced by cultivating microalgae.From fermentation tank, flask or its Harvesting microalgae biomass in allogene reactor.

In accordance with GMP programs.Whoso, if suffering from or doubtful suffering from by medical inspection or by monitoring observation proof Disease, open lesion (including furuncle, sore or wound of infection) or with any other abnormal microorganism pollution source, thereby It there is food, food-contacting surface or the contaminated obvious possibility of food packaging materials, then he will be prohibited to be expected Any operation of this pollution can be caused, until this situation is corrected.Instruction personnel will report to their person in charge This kind of health status.All staff directly contacted with microalgae biomass, biomass contact surface and biomass packaging material To prevent from observing hygienic habit in the degree needed for pollution in protection microalgae biomass when being on duty.The method for keeping cleaning Including but not limited to：(l) dress is suitable for protecting biomass, biomass contact surface or biomass packaging material to prevent the side of pollution The coat of formula operation.(2) appropriate personal cleanliness is kept.(3) left before start-up operation, every time after workbench and When any other time that hand is possible to be dirty or polluted, thoroughly washed one's hands in appropriate hand-washing facilities (if any must Protected to prevent the pollution of undesirable microorganism, then also to be sterilized).(4) remove and be possible to fall into biomass, equipment or appearance All unstable jewellery and other articles in device, and remove with hand operate biomass during can not adequately disinfected hand Decorations.If this kind of jewelry can not be removed, can use certain material covering jewelry, the material can keep completely, clean and defend Raw condition, and can effectively protect biomass, biomass contact surface or biomass packaging material not to be polluted by these articles. (5) gloves (if they are used in biomass processes) are kept in complete, cleaning and hygienic state.Gloves should be not permeable Saturating material.(6) in the appropriate case, hairnet, hair filler, cap, beard set are put in an efficient way or other are effective Beam hair care product.(7) clothes or other personal belongings are stored in beyond biomass exposure or washing facility or utensil place Place.(8) place in addition to following behavior being limited in the place of exposed except biomass or washing facility or utensil：Eat Biomass, chew gum, have a drink or smoking.(9) any other necessary precaution measure is taken to protect biomass, biomass Contact surface or biomass packaging material not by microorganism or foreign substance (include but is not limited to sweat, hair, cosmetics, tobacco, Chemicals and the medicine being applied on skin) pollution.Clasmatosis program optionally is carried out to microalgae biomass molten to produce Solve product and/or optionally be dried to form microalgae biomass composition.

Example 4

Chlorella protothecoides are cultivated to produce high oily algae piece

Using the process described in example 2 and 3, Chlorella protothecoides (UTEX 250) are produced with 5,000L fermentation tanks biological Matter.Glucose (corn syrup) concentration is monitored during whole experiment.When concentration of glucose is low, added into fermentation tank More glucose.After all nitrogen exhausts, cell starts to accumulate lipid.During whole experiment, take biomass samples to supervise Lipid content is surveyed, experiment is stopped when biomass reaches desired lipid content (by dry cell weight basis more than 40% lipid). In this case, harvested when biomass, which reaches, contains about 50% lipid by dry cell weight basis.

For microalgae biomass is processed into algae piece, the Chlorella protothecoides biomass of harvest and culture medium are divided using centrifugation From, and be dried using standard method on about 150 DEG C -170 DEG C of drum dryer.Roller drying obtained by packaging Chlorella protothecoides biomass (there is the lipid by dry cell weight basis about 50%, high lipid) and store and be provided as algae piece and make With.

Example 5

There is no Algae toxins in dry Chlorella protothecoides biomass

Grown using the method described in example 4 and be prepared for Chlorella protothecoides (UTEX 250) biomass samples.Using Liquid chromatography-mass spectrography/mass spectral analysis (LC-MS/MS) analyzes the algae and bluish-green Algae toxins in dry biomass with the presence or absence of pollution. These analyses cover all algaes mentioned in food regulations announced in document and international and bluish-green Algae toxins.As a result show, The biomass samples can detect that horizontal any tested algae or bluish-green Algae toxins without any.As a result it is summarized in table 3 In.

The algae of table 3. and the LC-MS/MS analysis results of bluish-green Algae toxins

Example 6

Fiber content in Chlorella protothecoides biomass

According to ACOC International official method (AOAC methods 991.43), to using example 4 and example 17 Described in method growth and prepare dry Chlorella protothecoides (UTEX 250) biomass sample carry out approximate analysis.To two Sample carries out the sour water solution of total lipid content (lipid/oil), and the fat content of high lipid algal biomass is about 50%, high protein The fat content of algal biomass is about 15%.The crude fiber content of both high lipid and high protein algal biomass is 2%.It is high The moisture (analysis is determined by weight) of both lipid and high protein algal biomass is 5%.By crucible calcination and inorganic Ash analysis determines content of ashes, and high lipid algal biomass is 2%, high-protein biological matter is 4%.By by each biology of calcination The nitrogen quantity of matter release determines crude protein, and high lipids, biological matter is 5%, and high-protein biological matter is 50%.Take fat, crude fibre, water Point, the above-mentioned given value of ash content and crude protein, its summation is subtracted from 100, by mathematic interpolation carbohydrate content.High lipid It is 36% that the carbohydrate of biomass, which calculates content, and the carbohydrate content of high-protein biological matter is 24%.

Further the carbohydrate content of two algal biomass of analysis shows there is about 4-8% (w/w) trip in sample From sugar (mainly sucrose).The multiple batches of algal biomass containing high lipid of test free sugar (detection fructose, glucose, sucrose, Maltose and lactose), sucrose amount is in 2.83%-5.77%；Germinatus sugar amount is from can't detect to 0.6%；Glucose amount is from detection Less than to 0.6%.Other sugared (that is, fructose, maltose and lactose) are not found in the batch of any measure.It is also tested for many The free sugar of the algal biomass containing high protein of batch, has only detected the sugarcane of 6.93% to 7.95% scope in any batch Sugar.

Two algal biomass are analyzed according to ACOC International official method (AOAC methods 991.43) Total dietary fiber (in the carbohydrate portions of algal biomass).The solubility that high lipids, biological matter contains 19.58% is fine The insoluble fibre of peacekeeping 9.86%, total dietary fiber is 29.44%.High-protein biological matter contains 10.31% Soluble Fiber With 4.28% insoluble fibre, total dietary fiber is 14.59%.

The Monosaccharide analysis of algal biomass

Use the monose methyl Portugal produced by sour methanol decomposition reaction (acidid methanologyis) from the sample The gas chromatography of the combination of complete-O- trimethylsilyls (per-O-trimethylsilyl, the TMS) derivative of polyglycoside/ Mass spectrography (GC/MS), has about 50% fat to what is grown and prepare using the method described in example 4 by dry cell weight basis The monose (glycosyl) of dry Chlorella protothecoides (UTEX 250) biomass samples of matter is constituted and analyzed.In short, leading to first Cross progress methanolysis in the 1M HCl at 80 DEG C in methyl alcohol and prepare methyl from dry Chlorella protothecoides in 18 to 22 minutes Glucoside, then with N acetylations (detection for being used for amino sugar) again of the pyridine and acetic anhydride in methanol.Then lead at 80 DEG C Cross to be handled with Tri-Sil (Pierce) and make the complete-O- trimethylsilylations of sample 30 minutes.These steps it is the previously described in Merkle and Poppe(1994)Methods Enzymol.230:1-15 and York et al. (1985) Methods Enzymol.118:In 3-40.It is being connected to using All Tech EC-1 fused silica capillary tubes post (30m x 0.25mm ID) The GC/MS analyses of TMS methyl glucosides are carried out on 5975b MSD HP 6890GC.By by their retention time and mark Quasi- condition ratio and identify these monose, and these carbohydrate characters are confirmed by their mass spectrum.Chlorella protothecoides Monose (glycosyl) composition be：1.2 moles of % arabinose, 11.9 moles of % mannose, 25.2 moles of % gala The glucose of sugar and 61.7 moles of %.These results are expressed as the molar percentage of total carbohydrates.

Example 7

The aminogram of algal biomass

According to AOAC International official method (Tryptophan analysis：AOAC methods 988.15；Methionine and Guang Propylhomoserin is analyzed：AOAC methods 985.28；Other amino acid：AOAC methods 994.12) to using the growth of method described in example 4 and Prepare, by dry cell weight basis have about 50% lipid dry Chlorella protothecoides (UTEX 250) biomass sample amino acid Content is analyzed.By the amino of the aminogram (being represented with the percentage of gross protein) of dry algal biomass and dry shell egg Acid spectrum (product specification table of the spectrum from shell egg, Protein Factory Inc., New Jersey) is compared, and is as a result shown Both sources have comparable proteinaceous nutrient value.The relative aminogram of the sample of Chlorella protothecoides is (relative to total egg Result in vain) shows that the biomass contains methionine (2.25%), cysteine (1.69%), lysine (4.87%), phenylpropyl alcohol Propylhomoserin (4.31%), leucine (8.43%), isoleucine (3.93%), threonine (5.62%), valine (6.37%), group Propylhomoserin (2.06%), arginine (6.74%), glycine (5.99%), asparatate (9.55%), serine (6.18%), Glutamic acid (12.73%), proline (4.49%), hydroxyproline (1.69%), alanine (10.11%), tyrosine And tryptophan (1.12%) (1.87%).The comparison of algal biomass and shell egg aminogram is shown in Figure 2.

Example 8

Carotenoid, phosphatide, tocotrienols and the tocopherol composition of the biomass of Chlorella protothecoides UTEX 250

The algal biomass that method described in use example 4 is produced is analyzed using positive HPLC, AOCS method Ce 8-89 The tocotrienols and tocopherol content of sample.Contain fertility triolefin using hexane or other non-polar solvent extract biomass The part of phenol and tocopherol.Complete tocotrienols and the result of the composition of tocopherol are summarized in table 4.

Tocotrienols and tocopherol content in the algal biomass of table 4.

Carotenoid is simultaneously analyzed using HPLC methods in the part containing carotenoid of separating biomass.In aluminium mortar The algal biomass (using the method production described in example 3) of freeze-drying is mixed with carborundum, with mortar and pestle Grinding four times, 1 minute every time, thus prepares the part containing carotenoid.Then grinding is rinsed with tetrahydrofuran (THF) The mixture of biomass and silicon simultaneously collects supernatant.Repeat to extract biomass, until supernatant is colourless, collect all extracts THF supernatants, and using standard HPLC methods analyze carotenoid content.Rolling is also used using above method analysis The carotenoid content for the algal biomass that cylinder drier is dried.

The carotenoid content of the algal biomass of freeze-drying is：Total lutein (66.9-68.9mcg/g:Wherein cis- Lutein scope is from 12.4 to 12.7mcg/g, and trans-lutein scope is from 54.5 to 56.2mcg/g)；Trans-luteole (31.427-33.451mcg/g)；Cis-luteole (1.201-1.315mcg/g)；T- α-kryptoxanthin (3.092- 3.773mcg/g)；T- beta-cryptoxanthins (1.061-1.354mcg/g)；15- cis-beta-carotenes (0.625-.0675mcg/g)； 13- cis-beta-carotenes (.0269-.0376mcg/g)；T- alpha-carotenes (0.269-.0376mcg/g)；C- alpha-carotenes (0.043-.010mcg/g)；T- beta carotenes (0.664-0.741mcg/g)；With 9- cis-beta-carotenes (0.241- 0.263mcg/g).The scope of the total carotinoid of record is from 105.819mcg/g to 110.815mcg/g.

The carotenoid content of the algal biomass of roller drying is significant lower:Total lutein (0.709mcg/g:It is wherein anti- Formula-lutein is 0.091mcg/g, and cis-lutein is 0.618mcg/g)；Trans-zeaxanthin (0.252mcg/g)；It is suitable Formula-zeaxanthin (0.037mcg/g)；α-kryptoxanthin (0.010mcg/g)；Beta-cryptoxanthin (0.010mcg/g) and t- β-carrot Plain (0.008mcg/g).The total carotinoid of record is 1.03mcg/g.These as shown by data, the method for drying algal biomass can Significantly affect the content of carotenoid.

Phosphatide analysis also has been carried out to algal biomass.Extracted using Folch extracting methods (chloroform, methanol and aqueous mixtures) Part containing phosphatide, oil samples are analyzed using AOCS official method Ja 7b-91, and HPLC determines the lecithin of hydrolysis (International Lecithin and Phopholipid Society1999), HPLC analysis phosphatide, scattering measuring (International Lecithin and Phospholipid Society 1995) method analyzes content of phospholipid.Total phospholipids It is 1.18% by w/w percentages.The Contents of Phospholipids of algae oil be phosphatidyl choline (62.7%), phosphatidyl-ethanolamine (24.5%), Lysophosphatidyl choline (1.7%) and phosphatidylinositols (11%).Contain phosphatide for what is extracted using hexane from algal biomass Part, has also carried out similar analysis.Total phospholipids is 0.5% by w/w percentages.Contents of Phospholipids be phosphatidyl-ethanolamine (44%), Phosphatidyl choline (42%) and phosphatidylinositols (14%).

Example 9

The food article of piece containing algae (high oil)

The heart/metabolic health rod

The composition of the heart/metabolic health rod is constituted：Instant oat (30.725%), crisp rice (9.855%), the sugared (sugarcane of particulate Sugar) (14.590%), light brown sugared (6.080%), salt (0.550%), Canola Oil (10.940%), corn syrup It is 42DE (7.700%), honey (3.650%), water (7.700%), lecithin (0.180%), sodium bicarbonate (0.180%), dry Algal biomass (lipids of Chlorella protothecoides UTEX 250,48%) (1.540%), corowise phytosterols (1.060%), inulin (Soluble Fiber) (4.280%) and psyllium (insoluble fibre) (0.970%).

Specification: (1) is in 325 degrees Fahrenheit convection current preheating ovens.(2) the one 5 kind of composition is weighed up in alms bowl.(3) in bar suddenly Mixing water, lecithin and sodium bicarbonate in special blender.(4) honey, corn syrup and Canola Oil are mixed；It is micro- Wave heating 30-40 seconds.Spatula hand mix is used, mixture is poured into Hobart's blender.(5) standard chow needed for adding Flavor enhancement.(6) dry nutriment (algal biomass, phytosterol, fiber) is added into Hobart's blender.(7) add surplus Under dry ingredients.(8) shape and bakeed 20-25 minutes in 325 degrees Fahrenheits to flowing down.

Cardio Daily Shot (liquid food containing the complete oily algal biomass of height)

The cardio shot of orange taste into being grouped into：Distilled water (869.858g), sodium benzoate (0.100g), The powder of Ticaloid 5415 (1.000g), dehydration sugarcane juice sugared (88.500g), the algal biomass (more than 40% oil) dried (16.930g), Fibersol-2ADM (47.000g), corowise ES-200 phytosterols (18.300g), granular citric acid (1.312g), orange extract (WONF, Flavor 884.0062U) (1.000g).Merge the composition and be blended until adjusting It is even.

Weight management smoothie (liquid food containing the complete oily algal biomass of height)

Smoothie based on fruit into being grouped into：Distilled water (815.365g), stabilizer (4.5g), cider concentration Thing (58g), orange juice concentrate (46.376g), lemon juice concentrate (1.913g), mango mud concentrate, banana puree (40.656g), passionfruit juice concentrate (8.4g), ascorbic acid (0.320g), algae piece (46.41g), orange flavoring extract (1g), pineapple flavoring (0.4g) and mango flavoring (0.16g).Merge the composition and be blended until mixing well.

The heart/metabolism piece (encapsulating/the oily algal biomass of complete height of tablet form)

Metabolic health piece into being grouped into：The dry microalgae biomass of Chlorella protothecoides (UTEX250, by dry cell weight basis More than 40% lipid) (1000mg/ pieces), Betatene beta carotenes (beta carotene 20% of Dunaliella) (15mg/ Piece), the vitamin C (100m g/ pieces) as ascorbic acid and Bioperine (pepper bioavilability reinforcing agent) (2.5mg/ Piece).

Algae snack chip

Algae snack chip into being grouped into：Bread flour (1 glass), mealy potato (1/2 glass), algal biomass are not bleached (by cell The lipid of dry weight meter more than 40%) (3 soupspoon), salt (3/4, to taste adjust), pearling cone meal (2 soupspoon), water (1/3-1 glasss) With flavoring (for example, fennel seeds, curry, pasture flavouring) (to taste).

Preparation process: mixing dry ingredients, 1/3 glass of water is added into dry ingredients.Add other water (at most 1 glass altogether) with Form dough.Dough is kneaded into uniform product, then allows to stand 30 minutes at room temperature.Cut the dough stood and formation Flakelet, bakees 20-30 minutes or is baked onto crisp at 275 °F.

Algae raisin cookie

Algae raisin cookie into being grouped into：Butter or margarine (1/2 glass, conventional 3/4 glass of recipe requirement), barley Piece or oatmeal (13/4 glass), nutmeg (1/4), water or milk (2-3 soupspoons), brown sugar (1 glass), salt (1/2), baking powder (1/2), vanilla (1), Chinese cassia tree (1), raisins (being optionally immersed in advance in brandy or orange juice) (3/4 glass) and Dry algal biomass (30% to oil) (1/3 glass).About 2 dozens of cookies are made in this recipe.

Conventional recipe requires 2 eggs and 3/4 glass of butter or margarine.Sleep with using dry algal biomass, pass through Substituted and avoided using 1/4 glass of butter or margarine and egg with the algal biomass of oil-containing.

Preparation process: butter and sugar is turned into creaminess.Stirring is untill quite soft.Add vanilla.Merge powder and Barley flake and algae.Merge butter mixture and powder-flake mixture.Add raisins.Flatten with teaspoon drip and somewhat. 375 degrees Fahrenheits are roasting about 9-10 minutes.

Algae barley pasta

Barley pasta with algae into being grouped into：Pearling cone meal (3/4), with by dry cell weight basis at least Algal biomass (2 soupspoon), big egg (1) and the salt (1/2) of the drying of 20% lipid.

Preparation process：Powder is put into alms bowl and algae and salt is added.Stir together.Centre plus egg (well shape is made) are simultaneously progressively mixed Enter powder.If it were difficult to admix, plus 1 soupspoon water would spray around.Start to knead dough when all powder have been mixed, be allowed to evenly. This process should be completed in 5-8 minutes.When dough is uniform, it is divided into two beads, olive oil is wiped in outside.Covering is simultaneously It is allowed to stand about 30 minutes.Dough is flattened, about 1/8th inches of thickness is then rolled to rolling pin (for as the wide face of Italy The pasta of sample).Pasta is cut into thin strips.It is put into the salt solution of boiling.About boil 8 to 10 minutes.It can use A small amount of broken Pa Masen cheese and some pepper corns, which are placed on the top of pasta, to be eaten.

Pasta

This example is compared (to be had by the high lipids, biological matter of full cell of the egg in conventional recipe and the conventional recipe of replacement The pasta being made by the Chlorella protothecoides (250 plants of UTEX) of dry cell weight basis 48%.

Table 5. eats the recipe of control for conventional pasta

Table 6. replaces the recipe of shell egg for full cell algal biomass

In each case, cooking step is:

1. hooked together in the precious alms bowl (kitchen aid bowl) in kitchen with dough and flour and salt.

2. gently beat eggs.Low speed (speed #2) adds the egg slightly broken up, until forming hard dough.

3. if desired, plus 1-2 soupspoons water stirring.

4. mixing 3-4 minutes, if dough is too viscous, plus a little extra flour.

5. a dough be divided into can tabletting several pieces.Allow to stand dough before tabletting 1 hour.

6. dough is pressed into desired thickness using pasta tablet press machine.

7. a pasta is cut into inch strips.

8. water flask is put on stove until boiling.

9. boiling pasta, stirred to prevent adhesion with oil/butter.Add sauce edible.

Whole-cell biological matter pasta has the texture and outward appearance for being similar to conventional recipe.Prominent algae taste is not Obviously.Full cell microalgae biomass improves the yield of dry pasta, it is likely that be due to water binding function The reason for.These observations can serve as the idea of good swelling agent with full cell algal biomass in dried foodstuff or processed food Unanimously.

Algae milk

Algae milk contain about 8% solid, its constitute have 4% beneficial heart lipid, 2.5% be rich in essential amino acid Protein, 1.5% carbohydrate and 0.5% fiber, and strengthened with vitamin A and D.Algae milk is very useful to be good for Health；It is vegetarianism, can be for instead of milk and soya-bean milk.From unlike milk, its saturated fat is very low, no As soya-bean milk, mainly monounsaturated (more than 50% C18 of fat:1).Algae milk taste is gentle；Unlike soya-bean milk " having fishy smell ".Can Add condiment, such as strawberry or raspberry.

Algae milk into being grouped into the dry full frustule (8%) containing about 40% lipid, vitamin D (200 unit) vitamin A (200 unit), xanthans (0.2%) and water (adding to 100%).By water heating with scattered xanthans.Then in the xanthans of temperature Disperse dry full frustule in solution and add vitamin.Then solution is homogenized with high pressure homogenisers and carries out Pasteur and gone out Bacterium.Here is the other preparation using algae powder.

Example 10

The production of algae homogenate (high lipid)

High lipid algae will be processed into using the method and the Chlorella protothecoides containing high lipid of conditioned growth described in example 4 Homogenate.For microalgae biomass is processed into algae homogenate, first by the Chlorella protothecoides biomass processing of harvest into algae piece (referring to reality Example 4).Then by dry algae piece with about 40% solid concentration rehydration in deionized water.Then used in 1000-1200 bars Algae piece suspension obtained by high pressure homogenisers (the GEA type NS1001) micronizing operated under stress level, until biomass Particle mean size is less than 10 μm.Resulting algae homogenate is packed and stored untill when using.

Example 11

Functional food product：It is homogenized in food as the high lipid algae piece and algae of fatty sub

Following instance description uses high lipid (more than 40 weight %) algae piece or algae to be homogenized as routine and low fat recipe In fatty sub.High lipid algae piece is prepared using the method described in example 4.Prepared using the method described in example 8 High lipid algae homogenate.

Chocolate Brownie

The chocolate Brownie that this example will be made of conventional recipe, low fat control recipe with high lipid algae piece (having the Chlorella protothecoides (250 plants of UTEX) by the lipid of dry cell weight basis 48%) is compared, to replace in conventional recipe Some fat.

Table 7. is directed to the recipe that conventional chocolate Brownie is compareed

Table 8. is directed to the recipe that low fat is compareed

Table 9. replaces the recipe of butter and egg for full algal biomass Brownie

In each case, cooking step is:

1. preheating oven is to 350 °F.Fat liquoring and powder is put on the baking trays of 8x 8.

2. melt butter and cocoa power in small pan.Shelve cooling.

3. beaten eggs in the precious alms bowl in attached vaned kitchen to foaming.It is gradually added sugar.

4. add room temperature/sl temperature butter/cocoa powder mixture into egg mixture.

5. flour is mixed with baking powder.Slowly plus 1/2 mixture is into pastel.

6. hickory nut is added in the remainder of powder.Plus mixture is into pastel.Mixed on low speed (speed #2), until Fully blending.Plus vanilla extract and mix.

7. a pastel is spread out in disk.It is roasting 20-25 minutes.

8. if desired, cool down Brownie and be allowed to freeze.

Compared with the Brownie made with algae piece or the Brownie of routine, low-fat control Brownie (save butter and Eggs) do not have same bits granular texture.Algae piece Brownie has good-looking visible bits granular texture, but than full-cream fat Blang Buddhist nun is somewhat finer and close and more more tacky.Generally speaking, compared with conventional Brownie, the fat content for the Brownie made with algae piece Reduce about 64%.

Sponge cake

This example is compared to be replaced in conventional recipe by conventional recipe, low fat recipe, high lipid algae homogenate (HL-AH) Eggs and butter and high lipid algae piece replace the sponge cake that the eggs in conventional recipe are made.Algae is homogenized and algae piece both of which is come From Chlorella protothecoides (250 plants of UTEX), with the lipid by dry cell weight basis 48%.

The conventional sponge cake recipe of table 10.

Table 11. is directed to the recipe of low fat negative control

Table 12. replaces the recipe of egg and butter for being micronized high lipid algal biomass

Table 13. replaces the recipe of egg for high lipid algae piece

In each case, cooking step is:

1. preheating oven is to 350 °F.Fat liquoring and powder is put on two disks of 9x 13.

2. powder, baking powder and sodium bicarbonate are sieved together.It is shelved on one side.

3. in the precious alms bowl in kitchen, butter and sugar are broken into creaminess until shinny together.Once buy 1 piece of egg.

4. add vanilla extract.

5. powder mixture and buttermilk are alternately added into pastel.It is mixed into and just combines.

6. a pastel is poured into the disk got ready.

7. roasted egg cake 35-40 minutes, or bake untill the toothpick of taking-up is clean.

8. cooling.

The sponge cake being made of high lipid algae piece (being used as the substitute of egg) is very fine and close, does not almost consider granular texture to be worth doing. However, compared with very fine and close and dry low fat negative control, the sponge cake being made of high lipid algae piece is moistening.With height The cake that lipid algae homogenate (HL-AH) (replacing butter and egg all in full-cream fat cake) is made is very wet and slippery in texture, And with the good bits granular texture for being similar to conventional recipe cake.During trial test, the cake being made of HL-AH is without conventional Butter taste in the presence of cake.Generally speaking, HL-AH is the good alternative of butter in conventional sponge cake recipe and egg. Cake with HL-AH contain than conventional sponge cake lean about 75%, but production cake there is the granular knot of good bits Structure, texture and humidity.

Biscuit

This example compares the eggs and shortening and high fat in routinely recipe, the conventional recipe of high lipid algae piece replacement The biscuit that eggs and shortening in conventional recipe are made is replaced in matter algae homogenate (HL-AH).Both algae piece and algae homogenizing biological matter Chlorella protothecoides (250 plants of UTEX) are all from, with the lipid by dry cell weight basis 48%.

The conventional biscuit recipe of table 14.

Table 15. replaces the recipe of egg and shortening for high lipid algae piece

Table 16. is homogenized the biscuit recipe of (HL-AH) using high lipid algae

In each case, cooking step is:

1. preheating oven is to 450 °F.

2. in the precious alms bowl in kitchen, merge powder, baking powder, sugar and salt.

3. shortening is added into mixture, until forming thick bits grain.(speed #2).

4. beat eggs and milk.Dry ingredients are added with moisture and is mixed, untill just dry ingredients are got wet.

5. mixing is until form dough (speed #2,15 seconds).

6. it is rolled to 3/4 " thick (or tabletting, if desired).With 2 1/2 of powder have been spread, " biscuit cutting knife is cut.

7. it is placed on the slightly baking tray of grease.Bake 8-10 minutes or bake to golden yellow.

8. eat while hot.

In terms of texture and outward appearance, the sample being made of HL-AH appears similar to full adipose versus.Generally speaking, HL-AH Biscuit maintains the texture of conventional recipe biscuit closest to the biscuit of conventional recipe, the biscuit lean 65% of generation With fermentation (rise).

Creaminess salad-dressing

This example is compared (has 20% fat using conventional recipe (control with 40% fat), low fat recipe Control) and be homogenized with high lipid algae (HL-AH) (have by weight about 20% fat) recipe mayonnaise/salad Baste, the high lipid algae homogenate comes from Chlorella protothecoides (250 plants of UTEX), with the lipid by dry cell weight basis 48%.

Table 17. is directed to the recipe of 40% adipose versus thing

Table 18. is directed to the recipe of 20% adipose versus

Table 19. is directed to the recipe of HL-AH creaminess salad-dressings

In each case, cooking step is:

1. merge yolk, acid, water and salt using food processor.

2. slow oiling, the emulsion until forming difficult demulsification.

If 3. the too difficult demulsification liquid of emulsion, adds some extra water.

4. scrape the material on lower edge and shear 10 seconds again to be incorporated to any oil droplet.

20% adipose versus flavouring (being made of Canola Oil) does not have any viscosity, and fails to form breast Liquid.Surface blistering foam, and form after flavouring is stood oil droplet.The flavouring being made of HL-AH has algal biomass Local flavor, good opacity and sticky and satiny mouthfeel.Generally speaking, compared with the flavouring of 20% and 40% fat, HL-AH imparts more preferable opacity and viscosity to flavouring.HL-AH plays important emulsifying agent effect, and with half Fat content produce the flavouring of the characteristic with 40% fatty flavouring (there is appropriate mouthfeel).With the HL-AH of micronizing (19% fat content) has obtained similar result in Dutch sauce recipe (fat that conventional recipe control is 80%).With The Dutch sauce of HL-AH productions is smooth and taste is abundant, with satiny mouthfeel and good viscosity.The color of sauce is than full-cream Fat control is somewhat dark yellow.Generally speaking, full-cream fat is comparable to by the product of the Dutch sauce production with micronizing HL-AH Tester, and lean 75%.

Model chocolate

This example compared with conventional recipe, (HL-AH) is homogenized with high lipid algae replace the milk and oil in conventional recipe with And the model chocolate nutritious drink that milk and oil in conventional recipe are made is replaced with high lipid algae piece biomass.Algae piece biomass Chlorella protothecoides (UTEX250 plants) are come from HL-AH both of which, with the lipid by dry cell weight basis 48%.

Table 20. is directed to the recipe of conventional chocolate drink control

Table 21. is directed to the recipe for the chocolate that milk and oil are replaced using HL-AH

Table 22. is directed to the recipe for the chocolate that milk and oil are replaced using algae piece biomass

In each case, cooking step is:

1) dry ingredients are blended.

2) added water point (in addition to a flavor enhancement) to kettle.

3) it is stirred in dry ingredients.

4) sheared 1 minute with rod agitator.

5) 200 °F are heated on furnace roof.

6) homogenized under 2500/500psi.

7) it is cooled to<40 °F and refrigerate.

Chocolate containing HL-AH has the outward appearance thicker, more rich than the chocolate of the piece containing algae, and outward appearance The upper chocolate closer to routine.Generally speaking, the HL-AH samples of micronizing are more closely similar to conventional chocolate beverage pair According to, good viscosity is imparted, and opacity is slightly above conventional chocolate drink control.

Example 12

The production of algae powder (high lipid)

Fermentation process and the Chlorella protothecoides containing high lipid of conditioned growth described in use example 4 is processed into high fat Matter algae powder.For microalgae biomass is processed into algae powder, the Chlorella protothecoides biomass of harvest is separated from culture medium, then Carry out centrifugal concentrating and be dried according to standard method using spray dryer.Algae powder obtained by packaging (has been sprayed dry The dry full frustule into powder type) and store untill when using.

Example 13

The production of algae powder (high lipid)

Fermentation process and the Chlorella protothecoides containing high lipid of conditioned growth described in use example 4 is processed into high fat Matter algae powder.In order to which microalgae biomass is processed algae powder, the Chlorella protothecoides biomass of harvest is centrifuged from culture medium.Make With high pressure homogenisers, ((GEA type NS1001) is operable to containing obtained by being micronized under the stress level of 1000-1200 bars The concentrated biomass of more than 40% moisture, until the particle mean size of biomass is less than 10 μm.Then it is even to algae using standard method Slurry is spray-dried.Algae powder (the micronizing frustule for being spray dried to powder form) and storage obtained by packaging are directly Untill when using.

Analyze the particle diameter of the sample of high lipid powders.A kind of algae powder in aqueous dispersion is produced, and is using Hydro 2000S annexesThe particle diameter of the algae powder is determined on the machines of Mastersizer 2000 using laser diffraction.Pass through Soft mixing produces a kind of control dispersion, and produces other using the pressure of 100 bars, 300 bars, 600 Palestine and Israels and 1000 bars Dispersion.The particle mean size of these results display algae powder is less under the conditions of with elevated pressures (in soft mixing condition Under be 3.039 μm, be 2.484 μm under the conditions of 1000 bars).Size distribution changes under the conditions of elevated pressures, wherein larger chi Very little (more than 10 μm) reductions of particle, and less particle (being less than 1 μm) increase.Distribution curve (the figure of soft mixing condition 5A), 300 bar conditions (Fig. 5 B) and 1000 bar conditions (Fig. 5 C) are as shown in Figure 5.Fig. 4 shows algae powder under an optical microscope Picture after being homogenized in aqueous dispersion soon.Arrow points to single algae powder particles (being less than 10 μm) and arrow is pointed to Agglomerating or cluster algae powder particles (being more than 10 μm).

Example 14

The food article of powder containing algae (high oil)

Following instance describe using high lipid (by weight at least 20%, typically 25-60% fat by weight Matter) algae powder is used as the fatty alternative in conventional recipe.Other example also demonstrates algae powder for prepared food (such as scrambled egg Powder) when improve moisture retention and improve texture unique function.Prepared using the method described in example 13 in following instance High lipid algae powder.

Chocolate Brownie

, the chocolate Blang that will of conventional recipe be made different using the function and taste spectral difference of high lipid algae powder in order to assess Buddhist nun is compared with the Brownie and the lipopenic Brownie of routine that are made of algae powder.Using high lipid (by dry weight About 53% lipid) algae powder replace butter and eggs.

The conventional Brownie recipe of table 23.

The lipopenic Brownie recipe of table 24.

The algae powder Brownie recipe of table 25.

In each case, baking step is：

1. preheating oven is to 350 °F.Fat liquoring and powder is put on 8 " x 8 " baking trays.

2. melt butter and cocoa power in small pan.It is shelved on while being cooled down.

3. together with an egg is beaten with vanilla, until somewhat spuming.It is gradually added sugar and remaining wet composition.

4. add butter/cocoa powder mixture to egg mixture.Merge remaining dry ingredients and be added slowly to wet mixture, directly To blending.

5. a pastel spreads out into disk the inside and bakeed 20-25 minutes, or bakees until sizing.

For the Brownie with algae powder, merge dry ingredients, algae powder is then added into dry ingredients.Then slowly wet Composition (water and vanilla) is blended into inside dry ingredients.Pastel is spread out into disk the inside and bakeed 27-28 minutes.

There is dry texture by the Brownie of conventional lipopenic recipe production, compared with for Brownie texture more like It is cake.The Brownie being made of algae powder (has the percent fat similar with the Brownie of lipopenic recipe, is about 8% fat) it is very wet and have with Brownie texture, but compared with conventional Brownie recipe (about 19% fat) More fragile bits granular texture.When with description in example 11 compared with the Brownie that algae piece is made when, be made of algae powder Brownie is less fine and close, with softer bits granular texture.Generally speaking, algae powder is butter and eggs in baked goods recipe Effective alternative, the product of production is in texture, taste and the product for being similar to conventional recipe in appearance.Algae powder, which is shown, to be made Unexistent unique function during with algae piece (for example, thinner bits granular texture, less gluing, texture is light).

The chocolate cake without seitan of individual size

Replace yolk and butter to prepare one kind not using algae powder (the 8% algae powder in water, so that a kind of slurry is made) The chocolate cake without flour containing seitan.The following component of the amount in bracket is used：Granulated sugar (130 grams)；Half sweet tea chalk Power (150 grams)；Water (20 grams)；8% algae slurry material (100 grams)；Salt (2.45 grams)；Baking powder (4.5 grams)；Vanilla extract (4 Gram)；And albumen (91.5 grams).By chocolate with hydration and and in the slow fusing waterborne of only micro-boiling.Then in room temperature It is lower that algae slurry agitation is added into chocolate mixtures.Then sugar (retaining 5 grams of sugar is used for albumen) and vanilla are added to the chalk In power mixture, then add baking powder and salt (retaining 0.15 gram of salt is used for albumen).Beaten eggs with medium speed white until spuming Untill, then add the salt of reservation.Then beat eggs white untill when forming soft bubble, then add the sugar of reservation.Then beat eggs Untill when being hard bubbled in vain until being formed.Then albumen is mixed into chocolate mixtures untill blending completely.Then by pasty state Thing injects the small mould of individual size and bakes 14 to 15 minutes (being rotated at 8 minutes) at 375 f)..It is this without seitan Chocolate cake without flour has the conventional texture and outward appearance without flour chocolate cake made of butter and yolk.At this Algae powder successfully instead of butter and yolk in individual preparation, for the chocolate cake without flour without seitan.

Mayonnaise

In order to assess the emulsifying capacity of algae powder, by the mayonnaise being made of algae powder with the mayonnaise that is made of routine recipe with And lipopenic mayonnaise is compared, the mayonnaise being made of algae powder is answered in water (40%w/v) It is former and homogenized under low pressure (100-200 bars) to produce slurry.Algae slurry material, which is used, has about 53% lipid by dry weight High lipid algae powder be made, and replace oil and yolk in conventional recipe completely.

The conventional mayonnaise recipe of table 26.

The conventional lipopenic mayonnaise recipe of table 27.

Table 28. is directed to the recipe for the mayonnaise being made of algae slurry material

In each case, step is：

1. merge acid, water and dry ingredients using food processor.

2. add yolk and be slowly injected into oil or algae slurry material.The emulsion of difficult demulsification should be formed.If emulsion is too difficult broken Breast, adds other water, until emulsion reaches desired denseness.

Dripped 3. scraping the material on lower edge and shearing again 10 seconds with being incorporated to any oil/slurry.

The viscosity for the mayonnaise being made of algae slurry material is in the viscosity of conventional mayonnaises and gluing for lipopenic mayonnaise Between degree.The mouthfeel of algae slurry material mayonnaise is comparable to conventional mayonnaise (but containing total fat less than 50%).In order to combine More water and (tighten) product that compacts are allowed to " can more spread out ", in lipopenic mayonnaise and algae slurry material yolk Instant food starch is required in both sauce.Generally speaking, all fat in conventional mayonnaises recipe are replaced using algae slurry material The mayonnaise that fat source (for example, oil and yolk) is produced has good viscosity and mouthfeel, is had no difference with conventional mayonnaise. Algae slurry material plays effective emulsifying agent effect, successfully instead of the function of the oil and yolk that see in conventional mayonnaises.

In other application, high lipid algae slurry material, which is used to make lipopenic honey mustard, dips in sauce/flavouring. The addition honey, mustard, white into the mayonnaise (slightly change and dip in the d spiss of sauce/flavouring to reach) prepared as described above Vinegar, lemon juice flavor enhancement and sea salt.Merge all compositions in food processor and be mixed into uniform smooth.Final products contain There is about by weight 14% algae powder, with about 8% total fat.The honey mustard of the powder containing algae, which dips in sauce/flavouring, to be had It is comparable to the satiny mouthfeel that conventional (full-cream fat) honey mustard dips in sauce.

Miso salad-dressing

In order to assess application of the algae powder in creaminess salad-dressing, using conventional recipe and being restored to slurry (40% Solid) the recipe containing high lipid algae powder be prepared for miso salad-dressing, be using institute in yolk sauce formulation above What the method stated was produced.

Table 29. is directed to the recipe of conventional miso salad-dressing

Table 30. is directed to the recipe for the miso salad-dressing being made of algae slurry material

In each case, dry ingredients are blended together, be shelved on one side.Water, vinegar and acid are blended together and put Put on one side.Respectively measure out miso.For conventional recipe, oils is merged and shelved.For containing The recipe of algae powder, respectively weighs up algae slurry material, oil and titanium dioxide and merges.Then water/vinegar is blended with high-shear mixer Mixture.After blending, dry ingredients are added in the water/vinegar mixture.Then with high-shear mixer be blended water/vinegar and Oil mixture is slowly injected into while dry ingredients.Then flavouring is heated to 190 °F and continues 2 minutes, then make flavouring It is moved through the colloid mill most closely set.Then bottled finished product flavouring and refrigerate untill when using.

Both conventional recipe and recipe containing algae powder produce thickness without transparent creaminess salad-dressing.In vision On, both flavouring are comparable in color and texture.Beans sauce salad-dressing contains about 30% fat, and uses algae powder The miso salad-dressing that slurry is made contains about 12.65% fat.Generally speaking, the Japan being made of algae slurry material The fat that beans sauce flavouring contains is less than the fatty half contained by the miso flavouring being made of conventional recipe, and Also maintain satiny mouthfeel and opacity.

Pizza dough/bread stick

Using conventional pizza dough/bread stick recipe and pizza dough/bread containing 5% or 10% algae powder by weight Rod recipe tests the ability that algae powder works in yeast containing dough application.The high lipid of pizza dough/bread stick of the powder containing algae Algae slurry material (40% solid) is made, and is produced using the method described in yolk sauce formulation above.

In each case, 7.3 grams of yeast are merged with 9.3 grams of Plain flours, and mixed with 58 grams of warm water.Yeast is mixed Compound stands at least 10 minutes at room temperature.In the sample of the material of slurry containing algae, in a mixer mix slurry and 167 grams of water Close, and merge with 217 grams of Plain flours and 4.9 grams of salt.In conventional recipe, only water and flour and salt are closed in a mixer And.After merging, yeast mixt is added into dough, 90 grams of Plain flours are added in addition.Then kneaded dough with hand, if dough It is overly wet, if desired, adding other flour again.Cover dough and be allowed to send out 1 hour in warm position.After fermentation, by face Group's portioning, or rolling is pizza dough, or it is configured to bread stick.Then dough 8-12 is bakeed in 450 °F of baking ovens Minute or bakee untill ripe.

The pizza dough and bread stick of conventional recipe are difficult to chew, with traditional crust.With the ratio of conventional recipe Sa dough is compared, and the pizza dough containing 5% algae slurry material has more like the same texture of crispbread, and more crisp.Contain The pizza dough of 10% algae slurry material is more crisp than the pizza dough containing 5% algae slurry material.In the bread being made of algae slurry material In rod, compared with the bread stick of conventional recipe, the bread stick for having 5% algae has the Chewy core of moistening.Containing 10% algae slurry material Bread stick is more moistened than the bread stick of 5% algae.The baking time of the bread stick of two kinds of powder containing algae all lengthens.Equally, powder containing algae There is atomic algae flavor enhancement in the bread stick of slurry, overall taste is not disturbed.Generally speaking, with the bread stick of conventional recipe Compare, algae slurry material improves the crispness of pizza dough, and give more like the same texture of crispbread, and add The humidity of bread stick.In another application, high lipid algae slurry material (40% solid) is used in a kind of corn-dodger recipe simultaneously And with being compared by the corn-dodger of conventional recipe making.Similar to pizza dough result, the corn-dodger containing algae slurry material exists More like crispbread in terms of texture, and it is more crisp than the corn-dodger of conventional recipe.

Cream roll

Yolk and butter are replaced using algae powder and are prepared for cream egg using the following component with the amount in bracket Roll up, these compositions are：Warm water, about 110 °F (54.77 grams)；Speed fermentation is female (3.5 grams)；The whole milk (58.47 boiled Gram)；Algae powder (45.5 grams)；Granulated sugar (10 grams)；Plain flour (237 grams)；Active wheat gluten (15 grams)；Salt (3.5 grams)；And egg In vain (42 grams).Yeast is sprinkling upon on warm water and 5 minutes are stood.The milk boiled is added when milk temperature reaches 110 °F to 115 °F Add in yeast soln and mix to be allowed to combine addition sugar and mix to be allowed to dissolve.Then plus algae powder and mix until It is completely combined.Remaining dry ingredients are merged, and yeast/milk mixture is added into these remaining dry ingredients.Then Albumen is added immediately in the mixture and mixed using food processors (10 times, handled per subpulse dough/pasta 1 to 2 times).Then the dough/pasta pulse is handled five times or more times, continues 3 to 5 seconds every time, if it is desired, the more water of addition. Finished product dough/pasta is soft and has slight viscosity.The dough/pasta is covered with cloth and small in warm place standing one When, and make its size doubles (being about 2 to 3 times of original size).Then with food processors pulse processing 2 to 3 again It is secondary, continue to expand it in 1 to 2 second every time, and allow to stand until its size doubles again.Then dough/pasta is turned to one On individual surface and flatten to remove air.Then it is the dough/pasta rolling is rectangular and roll, by its border seal.Then The dough/pasta is placed in a disk, and allows to leaven dough again until its size doubles, it is placed in a pre-add Heat is into 400 °F of baking oven and roasting about 35 minutes.The cream roll has the outward appearance and texture of conventional cream egg roll, and Represent using algae powder without butter or the successful formulation product of the cream roll recipe of yolk.

Bread without seitan

Algae powder is tested in the yeast doughs bar without seitan containing the bread without seitan of algae powder by preparing The ability worked under part.Because without seitan and no wheat, algae powder is suitable for being incorporated into seitan and/or wheat mistake The diet of the crowd for quick disease/do not tolerate.Following component has the amount in bracket：Middle muscle is free of the powder mixture (3 glasss) of seitan, its It is configured to：2 glasss of sorghum flours, 2 glasss of brown rice ground rice, 1.5 glasss of farinas, 0.5 glass of glutinous rice powder, 0.5 glass of glutinous rice flour, 0.5 glass of wood Potato powder, 0.5 glass of three-coloured amaranth powder and 0.5 glass of quinoa powder；Dried milk (1/3 glass)；Guar gum (2)；Xanthans (1¹/₄Tea Spoon)；Tasteless gelatin or agar powder (1¹/₂Teaspoon)；Sugared (3)；Salt (1)；Egg substitute (1¹/₂Teaspoon)；Bread ferment Female (1 bag or 2¹/₂Teaspoon)；Shell egg (2 pieces)；Butter (5 soupspoons, be cut into small pieces)；Water or common soda water (1¹/₂Cup)；Honey (1 Soupspoon)；And apple vinegar (1).By bread ovenware somewhat grease and spread glutinous rice powder.By these dry ingredients in mixing Mixed in alms bowl untill thoroughly blending.Egg, butter, vinegar and honey are blended in one big alms bowl, then by 1 glass of water or soda water Add in the egg mixture.These dry ingredients mixed are slowly merged with egg mixture.Remaining water is slowly added to, and And then add remaining dry ingredients, then mix untill the pastel reaches the denseness of thick cake pastel.So Afterwards by this pastel with mixed at high speed about 5 minutes.Then the pastel is injected in the whole piece bread ovenware and covered, Then sent out 1 hour in warm position.Then the dough/pasta is baked 55 to 60 points in a baking oven for being pre-heated to 375 °F Clock, after 15 minutes with paper tinsel cover to prevent the excessive brown stain of crust.Then bread is removed from baking oven immediately, and in cutting It is preceding to be cooled down completely on grill frame.The bread without seitan has the outward appearance and texture of conventional strip bread.This demonstrate that algae Success of the powder in the yeast doughs application without seitan is used.

Soft baking chocolate dragee cookies

Use conventional soft baking chocolate dragee cookies recipe, lipopenic soft baking chocolate dragee cookies recipe and use The chocolate dragee cookies of high lipid algae slurry material (using what is produced with above identical method described in yolk sauce formulation) making Test the ability that algae powder works in cookies application.In both conventional cookies recipe and lipopenic cookies recipe, Algae slurry material also substituted for all butter and eggs.

Table 31. is directed to the recipe of conventional soft baking chocolate dragee cookies

Table 32. is directed to the recipe of lipopenic soft baking chocolate dragee cookies

Table 33. is directed to the recipe of the soft baking chocolate dragee cookies with algae slurry material

In each case, step is：

1. preheating oven is to 350 °F.Merge powder, sodium bicarbonate, baking powder and salt in alms bowl.It is shelved on one side.

2. butter/algae slurry material is broken into creaminess untill smooth with sugar and corn syrup.Squeeze into egg (if Words) and vanilla.

3. it is gradually added dry ingredients and mixes, until just forms dough.Admix chocolate dragee.

4. take a few soupspoon doughs；It is allowed in cookies baking sheet or rolls into ball and be placed in cookies baking sheet.

5. bakeing 16-18 minutes or being baked onto golden brown, cookies baking sheet is rotated in the whole midway that bakees.

Conventional recipe cookies are spread out well during bakeing, and soft when from baking oven out.For lipopenic Cookies, first dough/pasta is spread out not open, and also so, dough is driven plain before baking for later batch.Lipopenic cookies It is soft when from baking oven out, through the cookies for being cooled into densification.Lipopenic cookies also have early stage corn syrup Obvious flavour.Algae powder cookies have spreading out the property for being similar to conventional recipe cookies during bakeing, and texture is better than fat and subtracted Few cookies.After lower three days of environment temperature, algae powder cookies are all more moistened than conventional recipe cookies and lipopenic cookies.Always For, algal biomass slurry can effectively replace the butter and eggs in cookies application.Functionally, algal biomass slurry extends The shelf-life of cookies, this is that cookies maintain more moisture at ambient temperature after three days.

The shelf-life research of oatmeal raisin cookie without seitan

Due to the result of the extension of shelf-life tested from above chocolate dragee cookies, using high lipid algae powder (by dry The lipid of restatement about 53%) the oatmeal raisin cookie without seitan is made, carried out using the method described in example 13 Production.Cookies are bakeed, are then kept for seven days at ambient temperature.Initial sensory testing is carried out to cookies immediately after bakeing and cooling down With water activity test.Other sensory testing and water activity test the 1st, carry out within 3 and 7 days.On the day of each test, one Cookies are cut into small pieces, and raisins and oat is uniformly distributed in the sample.Each cookies at least wants two in water activity test Individual sample, the accuracy to ensure measurement.Aqua Lab, 3TE ranges of models (Decagon is used according to the scheme of producer Devices, Inc) instrument come carry out water activity (Aw) test.Briefly, water activity measurement is vapour pressure, its basis weight products In useful non-chemical combination water；Aw values are higher, then product is more moistened.In the application of this cookies, higher Aw values and compared with The long shelf-life is associated.0.65 Aw levels are desired targets.

Table 34. is directed to the recipe for the oatmeal raisin cookie without seitan being made of algae slurry material

Step is：

1. preheating oven is to 375 °F.

2. the dry ingredients in addition to oat and algae powder are blended together.Make oat aquation in 1/4 water.In 3/4 water In make algae powder aquation, and be fully blended using hand-held blender.Allow oat and algae powder aquation 10 minutes.

3. adding the algae powder of aquation into dry ingredients, it is sufficiently mixed.Add vanilla simultaneously be sufficiently mixed, until blending and it is smooth Untill.

4. it is just uniform to add oat and grape dry doubling to be mixed into.

5. cookies portioning is each gently pushed in cookies baking sheet.

6. cookies are bakeed in an oven 20 minutes, it is whole to bakee midway rotation cookies baking sheet.

Sense organ and water activity test result are summarized in table 5 below.The assessment of sensory testing's sample uses ten point system：1-2= It can not receive；3-4=is poor；5-6=is fine；7-8=is good；9-10=is excellent.Generally speaking, when at ambient temperature keep seven days When, the dessert prepared with algae powder maintains good humidity level, and taste and texture are rotten seldom.

Table 35. sensory evaluation scores of oatmeal raisin cookie and water activity result at ambient temperature.

Scrambled egg (comes from powdered egg)

Algae powder water conservation is tested in powdered egg application is restored and the ability that texture improves is provided.Egg is prepared using conventional recipe Powder, with the alternative of the high lipid algae powder of different content (5%, 10% and 20%) as the powdered egg of corresponding percentage (w/w).With The algae powder used in lower formula is prepared using the method described in example 13, and contains by dry weight about 53% lipid.

Table 36. is directed to the conventional recipe of the scrambled egg from powdered egg

Table 37. is directed to the scrambled egg recipe from the powdered egg with 5% algae powder

Table 38. is directed to the scrambled egg recipe from the powdered egg with 10% algae powder

Table 39. is directed to the scrambled egg recipe from the powdered egg with 5% algae powder

In all cases, egg prepare it is as follows:

1. algae powder (if any) is mixed with powdered egg.Egg is mixed with water.Stirring is until smooth.If desired Words, any agglomerate is sheared using hand mixer.

2. pour into egg mixture in the non-stick cooker of preheating.

3. boiled egg mixture is until shape, and be seasoned as desired.

All formulations color is similar, does not have significant color distortion between the egg of conventional recipe and the egg of the powder containing algae.It is conventional The egg of recipe be in texture it is dry, excessively inflate, it is soft, and lose satiny mouthfeel.With 5% microalgae biomass system Standby egg is more moistened and more solid in texture than the egg of conventional recipe.Mouthfeel is more satiny than conventional recipe egg.Use 10% algae Egg prepared by powder will also be moistened more than conventional recipe egg, and with the texture and mouthfeel of the scrambled egg prepared by fresh hen egg.With The overly wet profit of egg prepared by 20% algae powder, the texture with half-cooked semi-liquid-like egg.Generally speaking, with the conventional powdered egg phase prepared Than including for algae powder improves mouthfeel, texture and the humidity of prepared powdered egg.In 5% and 10%, algae powder is applied in egg Well, fat content is not dramatically increased.At 20%, algae powder gives too many moisture, makes the texture of powdered egg of preparation not It can receive.

Powdered egg keeps test

Because algae powder can increase significant moisture and improve the texture of powdered egg, ripe egg is maintained at steam table in order to assess How is performance when upper, has carried out following holding test.The scrambled egg being made of conventional recipe before furnace roof boiling is placed in water Change 10-15 minutes, the conventional recipe is using powdered egg, 5% algae powder and 10% algae powder (being all made of method as described above). After cooked, sample is immediately transferred in steam table, there covered them 30 minutes, temperature is at 160 to 200 °F.Every 10 Minute makes fresh sample with keeping sample to be compared.Sample evaluating uses ten point system：1-2=can not receive；3-4=is poor；5-6= Still；7-8=is good；9-10=is excellent.Test result is summarized in table 4 below 0.

The powdered egg of table 40. keeps the sensory results of test

UseTest the ability that algae powder improves the white texture of scrambled egg and mouthfeel.100 grams are fried with small non-sticky frying panAbout 1-2 minutes, untill egg is shaped.Without using butter or flavouring.Sample has the height that 10%w/w is replaced Lipid algae slurry material (is prepared, algae powder is containing by dry weight about 53% fat using the method described in as above being applied in mayonnaise Matter).With algae powderPreparation method it is identical with reference examples.

Control sample has the denseness more like water, the same in orally-dissolvable, the relatively little or no texture of texture more like water. The sample containing 10% algae slurry material quickly made is more like the scrambled egg being made of fresh egg.The sample of 10% algae slurry material also has More scrambled egg texture, and with plentiful mouthfeel, similar to the mouthfeel for the scrambled egg being made of fresh egg.Generally speaking, add Algae slurry material improves the white texture and mouthfeel of scrambled egg very successfully, albumen is tasted more like the stir-fry being made of fresh shell egg Egg.

Egg pulp

Algae powder is tested in research is kept improves the texture and the ability of moisture of the scrambled egg using egg pulp, and using sense Official group.Prepare egg pulp as control according to manufacturers instructions, and with 10% algae slurry material (2.5% algae powder and 7.5% Water) prepare egg pulp compare.The egg of control and 10% algae powder is all quickly made scrambled egg, totally 60 points are kept in steam table Clock.Take the sample of each scrambled egg product and every 10 minutes in sense organ group are tested.Sense organ group is judged by 1 to 9 point of system Overall appearance, humidity level, texture and the mouthfeel of scrambled egg product, 1 is can not receive, and 3 can not receive for appropriateness, 5 be it is fine, 7 It is acceptable, 9 be excellent.

Generally speaking, 10% algae slurry material of addition (2.5% algae powder solid) improves the texture of prepared egg, humidity water Gentle mouthfeel.In steam table after 60 minutes, compared with compareing scrambled egg, the scrambled egg product with 10% algae slurry material is still subjected to (sensory evaluation scores are 5), control scrambled egg can not be then being received in the unacceptable scope (sensory evaluation scores are 2.7) of appropriateness.It is all The result at time point is summarized in Fig. 3.

Pancake with powdered egg

It was found that the pancake of retail shop/waffle powder contains shell egg meal component.As shown in above powdered egg formula, high fat is added Matter algae powder improves the texture and mouthfeel of prepared egg products.Testing high lipid algae powder improves with i.e. mixed type pancake powder system Into pancake texture and the ability of mouthfeel.

Table 41. compares the recipe of pancake

Table 42. is directed to the recipe of the pancake containing high lipid algae powder

In both cases, rehydration powdered egg, algae powder and non-fat solid are come using water.Then remaining composition is added simultaneously It is stirred to pastel smooth.In each several part for pastel being poured into the pancake size inside the non thermal bonding pot of non-fat liquoring.Do thin Flapjack, until top bubbles burst, then twists over, until cooking.

Two kinds of pastel outward appearances are similar, do two kinds of pancakes and all take about the same time quantum.The thin of the powder containing algae is baked Cake than control pancake in texture it is lighter, more satiny and more fluffy and elastic smaller.Generally speaking, 50 weights are replaced with algae powder Amount % dried whole-egg produces the more preferable pancake of texture, and with more preferable mouthfeel.

Algae milk/frozen dessert

The other formula of algae milk has used high lipid algae powder.The algae milk contains following component (by weight): 88.4% Water, 6.0% algae powder, 3.0% whey protein concentrate, 1.7% sugar, 0.6% vanilla extract, 0.2% salt With 0.1% stabilizer.Merge each composition and homogenized under low pressure using hand-held homogenizer.Obtained by being cooled down before edible Algae milk.Mouthfeel is comparable to full milk, and with good opacity.Algae powder used contains about 50% lipid, therefore gained The algae milk arrived contains about 3% fat.When compared with vanilla flavored soymilk (Silk), algae milk has similar mouthfeel and opaque Property, without the beany flavor of soymilk.

Algae milk and other sugar and vanilla extract are then combined with, and 2-4 minutes are mixed until uniform in agitator. Mixture is put into the ice cream producing machine of precooling (Cuisinart) 1-2 hours, until reaching desired denseness.Prepare and use 220 gram 2% of milk and 1 yolk half and half the totally 325 conventional recipe ice creams restrained as comparing.Conventional recipe ice cream The denseness having is comparable to soft diet ice cream, is the ice cream that taste is abundant, texture is smooth.Although the ice cream being made up of algae milk Lack the careful sense (creaminess) of entirety and the mouthfeel of conventional recipe ice cream, but denseness and mouthfeel are comparable to what taste enriched Milk freezes.Generally speaking, the use of algae milk is successful in frozen dessert application：The frozen dessert algae milk of generation is regular ice cream Low fat substitute.

Orange algae beverage

A kind of orange taste algae beverage will use following component (being used in the amount in bracket) to prepare：Distilled water (879.51 grams)； Granulated sugar (30 grams)；Salt (1.9 grams)；Algae powder (50 grams)；Carrageenan (0.14 gram)；The stabilizers (0.75 of FMC Viscarin 359 Gram)；Vanilla extract (6 grams)；Lactalbumin (Eggstend) (30 grams)；And flavoring orange essence (1.7 grams).By these into division And and homogenized with a collection of homogenizer under 300 bars 1 time.The orange algae beverage is let cool and then edible.The drinks taste It is similar to dreamcicle, and very smoothly and with satiny mouthfeel, similar to whole milk, although it, which only contains, presses The fat of weight in wet base meter 2.5%.

Without egg eggnog

It is a kind of without egg eggnog following component (being used in the amount in bracket) to be used to prepare：Distilled water (842.5 grams)；Granulated sugar (50 grams)；Salt (2.3 grams)；Algae powder (50 grams)；Carrageenan (0.2 gram)；The stabilizers of FMC Viscarin 359 (1.0 grams)；It is fragrant Careless extract (3 grams)；Lactalbumin (Eggstend) (50 grams)；And nutmeg (1 gram).These compositions are merged, and used A collection of homogenizer homogenizes 1 time under 300 bars.The eggnog is let cool and then cooling is eaten.The eggnog has the outer of conventional eggnog See and mouthfeel, but due to lacking yolk and thick cream in recipe, its fat content (based on weight in wet base 2.5% fat) has shown Write reduction.

Cheese flavourings juice

A kind of cheese sauce will use following component (being used in the total weight percent in bracket) to prepare：40% algae slurry material (65.9%)；Xanthans (0.22%)；Pure starch slurry (Pure flow starch) (0.81%)；Water (26.6%)；Sugar (0.25%)；Salt (0.54%)；50% acetic acid (0.5%)；The cheese powder (5%) of enzyme modification.These compositions are mixed Untill smooth.This is the successful proof that algae powder is used for during tasty cheese sauce is applied.

Alginic acid milk

A kind of Yoghourt will use following component (being used in gross weight (500 grams) percentage in bracket) to prepare：Algae powder (1.25%)；Skimmed milk (50%)；Sugared (1%)；Salt (0.1%)；Deionized water (47.15%)；And starter culture (0.5%).Used starter culture is Euro culinary art Yoghourt starter cultures (Euro Cuisine Yogurt Starter Culture), it is (lactic acid (L.bulcaricus), thermophilic that it contains skimmed milk power, sucrose, ascorbic acid, lactic acid bacteria Hot streptococcus (S.thermophilus) and lactobacillus acidophilus (L.acidophilus)).Will except the starter culture it Outer all the components, which merge and are heated to 185 °F, continues 5 to 10 minutes, is then cooled to 105 °F to 110 °F using ice bath. Then the starter culture is added in the cooling yoghourt mixture, and in the domestic machine for making sour milk of Waring Pro YM 350 It is middle to be incubated about 8 hours.It is sour that the Yoghourt, which is tasted, shows that using the fermentation process of the starter culture of the work be success 's.The denseness of Yoghourt is soft and than boruga slightly stiff.

Extra experiment has been carried out in tasteless fat-free Yoghourt, and algae powder is combined to determine it to no fatty acids The contribution of the mouthfeel of milk.5 percent (by weight) algae powder is blended and enters a kind of tasteless fat-free Yoghourt (Pavel) In until smooth and untill preferably combining.The Yoghourt is let cool and then edible again.This contains the fat-free Yoghourt of 5% algae powder (it contains about 2.5% fat now) is with such as full-cream tasteless Yoghourt (Pavel) control, (it has 3.5% fat Content) equally plentiful and satiny mouthfeel.

Example 15

Algae oil

The solvent extraction oil from biomass

Algae oil is extracted from microalgae biomass, the preparation of the microalgae biomass is as described in example 1-4, using draping over one's shoulders herein The method dried biomass of dew, biomass is crushed using method disclosed here, makes broken biomass and organic solvent (for example Hexane) contact time enough section, to allow oil and hexane formation solution.Then filtering solution and oneself is removed by rotary evaporation Alkane is to reclaim the oil of extraction.

The solvent-free extract oil from biomass

Algae oil is extracted from microalgae biomass, the microalgae biomass is prepared as described in example 1-4, dried biomass, Physics crushes biomass in oily seed squeezer, and wherein algae oil is discharged from biomass.Then reclaim so from broken Biomass in the oil separated.

From algal biomass supercritical fluid extraction oil

Extracted using such as the supercritical fluid extraction (SFE) described in example 1-4 from Chlorella protothecoides (UTEX 250) Microalgae oil.The sample of the microalgae biomass (25.88 grams) is charged in extraction vessel, and make CO₂Gas is (in the pressure of selection Under power and temperature conditionss) continued for some time by the container until the gas of desired gross mass has been by the container Only.Then the high-pressure gas flow and the material of the extraction is made to enter by a pressure-reducing valve comprising these extractables (algae oil) In collector.After desired amount of gas has flowed through the extraction vessel, the collector is removed.Receive after extraction Collection retains the material (or residue) retained in this embodiment.15.68 grams of algae oil is extracted, and the residue weighs 10.2 Gram.The residue includes the algal biomass of degreasing, and with a kind of white powdered outward appearance.

Method using standard HPLC methods and described in example 8, to being analyzed using the SFE algae oils produced, Analyze antioxidant (12.7ppm TBHQ (TBHQ)), chlorophyll (1ppm), free fatty (1.34%), Karl Fischer humidity (0.05), monoglyceride (0.04%), diglyceride (2.52%), phosphatide (below nothing-detection level), life Educate phenol and sterol and tocotrienols.The algae oil includes following tocopherol and sterol：Methyltocol (0.13mg/100g)；γ gives birth to Educate phenol (0.20mg/g), alpha tocopherol (5.58mg/100mg)；Ergosterol (164mg/100g)；Campesterol (6.97mg/ 100g), stigmasterol (6.97mg/100g)；Cupreol (5.98mg/100g)；And 176mg/100g other sterols.The algae Oil also includes 0.24mg/g α tocotrienols.

The diversity of lipid chain in algae species

The lipodogramme of the lipid samples of the hypotype of the strain grown in example 1 is analyzed using HPLC.As a result it is shown in Fig. 1 In.

Example 16

Dietetic product and food article containing algae oil

Algae oil capsule (the bag that (a) is extracted by way of solvent extraction or (b) are extracted by non-solvent from algae The oil of envelope)

Tocotrienols, tocopherols, carotenoid, Ω -3 and the sterol that comprehensive protection system-offer naturally occurs Algae oil.It provides the use that the inanimacy substitute based on plant substitutes fish oil.

The composition of the Exemplary nutritive item compositions of table 43.

Algae oil (oil extracted solvent extraction or by way of being extracted by non-solvent from algae)

The composition of the Exemplary nutritive item compositions of table 44.

Brownie and spice cake containing algae oil

Used in baked goods application from the Chlorella protothecoides (UTEX grown by the fermentation process described in example 4 250) oil extracted in.Sponge cake (Moist Deluxe, Duncan Hines) and Brownie are made using 1/3 glass of oil (Chocolate Chunk, Pillsbury), the oil is that the instruction advised according to producer is extracted from Chlorella protothecoides.Chicken The result of both cake and Brownie and the sponge cake and cloth made using vegetable oil and identical boxlike mixture (box mix) Lang Ni is without what difference.

Example 17

The production of high protein algal biomass

The Heterotrophic culture of microalgae with high protein content

The Chlorella protothecoides (UTEX 250) that different oxygen is produced are grown under the conditions of rich nitrogen, the condition is by following one kind Or many kinds of substance offer：Yeast extract (organic nitrogen source), NH₄OH and (NH₄)₂SO₄, the culture described in supplement example 2-4 Base.In addition to culture medium, fermentation condition is identical with the condition described in example 2.After the exponential increase of about 3-5 days, when it reaches To harvest high protein algal biomass during desired culture density.Any of above processing method (the algae piece in example 4, reality Algae powder in algae homogenate in example 10, example 12, and the algae powder in example 13) it can apply to high protein microalgae biology Matter.

The approximate analysis of microalgae biomass

High-protein biological matter is processed into using the method described in example 4 by algae piece.To dry biomass, high lipid (example 4) and high protein all analyzes moisture, fat, fiber, ash content, crude protein and protein digestibility, using according to ACOC International official method.As a result it is summarized in table 4 below 5.

Table 45. has the approximate analysis of the microalgae of high protein content

ND=is not done

Calculate the difference of total carbohydrates：100% subtracts the known percentage in approximate analysis.High lipid by weight The total carbohydrates of biomass are about 36%, and the total carbohydrates of high-protein biological matter by weight are about 24%.

Crude fibre above represents the amount of the cellulose and lignin (in addition to other components) in biomass samples.Using According to ACOC International official method (AOAC methods 991.43), Soluble Fiber is carried out to biomass and insoluble The measurement of both property fibers (being exactly total dietary fiber altogether), both belongs to the carbohydrate ingredient of biomass.For High lipids, biological matter, Soluble Fiber is 19.58%, and insoluble fibre is 9.86% (total dietary fiber is 29.44%).For High-protein biological matter, Soluble Fiber is 10.31%, and insoluble fibre is 4.28% (total dietary fiber is 14.59%).

Using the method stated in example 8, it (is two batches grown as described above also to analyze high-protein biological matter Biomass) two samples (sample A and sample B) chlorophyll, sterol, tocopherol and tocotrienols.For sample A's Result is：Chlorophyll (93.1ppm)；Total sterol (1.299g/100g) it include：Cholesterol (1.05mg/100g), brassicasterol (301mg/100g), ergosterol (699mg/100g), campesterol (13.8mg/100g), stigmasterol (15.7mg/100g), And cupreol (3.72mg/100g), other sterols (265mg/100g)；Alpha tocopherol (0.18mg/g)；And α fertilities three Alkene phenol (0.03mg/g).Result for sample B is：Chlorophyll (152ppm)；Total sterol (2.460g/100g) it include：Courage Sterol (1.01mg/100g)；Brassicasterol (549mg/100g)；Ergosterol (1.39g/100g)；Campesterol (22.6mg/ 100g)；Stigmasterol (26.1mg/100g)；Cupreol (2.52mg/100g)；And other sterols (466mg/100g)；It is total raw Educating phenol (0.79mg/g) includes：Alpha tocopherol (0.35mg/g), gama tocopherol (0.35mg/g) and methyltocol (0.09mg/ g)；And α tocotrienols (0.01mg/g).

Protein digestibility in microalgae biomass

(0.2% Pepsin digestibility is tested, AOAC methods 971.09) is tested using Vitro Digestibility, to multiple batches of High protein and high lipids, biological matter (using the method production described in example 4) and high-protein biological matter analysis digestibility.It is right In high lipids, biological matter, total crude protein percentage range is 5.4% to 10.3%, wherein the percentage of total digestible protein Than scope 46.4% to 58.6%.For high-protein biological matter, total crude protein percentage range 40.8% to 53.3%, The percentage range of wherein total digestible protein is 71.6% to 85.3%.The biological meals also extracted to hexane (biomeal) (the high lipid algal biomass after the extraction of algae oil hexane) carries out same digestibility experiment.All batches of test In, total crude protein percentage is about 11-12%, wherein total digestible protein percentage range 76.72% to 80.2%.

When (total crude protein percentage is about 40.9%, and total digestible protein is 95.35%) phase with full soy meal Than when, total digestible protein percentage of high protein algal biomass is slightly less than full soy meal.Algae is thin to be caused to machined The high protein microalgae biomass that born of the same parents are mainly dissolved has carried out other experiment.These experiment as a result, total digests egg White matter percentage is comparable to full soy meal (about 95% total digestible protein).Generally speaking, high protein is biological The total crude protein percentage of matter and total digestible protein level of percent are comparable to full soy meal.

The digestibility result of the test for the biological meals that hexane is extracted shows that biological meals are as the additive of animal feed Feasible.Biological meals have remaining protein and oil concurrently, and with the percentage of about 80% total digestible protein Level.

Example 18

The food article of the algal biomass containing high protein

Use the food ingredients composition of high protein algal biomass (algae piece and algae are homogenized)

The high protein algal biomass used in the recipes below was produced with the methods described in Example 17above._<}95{>Given birth to the method described in above example 17 Produce high protein algal biomass used in following recipe.Algal biomass in following recipe comes from Chlorella protothecoides UTEX 250, it contains by weight about 51% protein, is referred to as high protein algal biomass, is appointed as algae piece or algae homogenate.

Vegetarian diet hamburger patti

This example compares vegetarian diet hamburger patti that routinely recipe is made, wherein with high protein microalgae biomass (algae Piece or algae homogenate (AH)) replace vegetable protein source (textured soybean protein (TSP), gluten and/or soy protein isolate (SPI))。

The conventional vegetarian diet hamburger patti recipe of table 46..

Table 47. is directed to be made of high protein algae piece replacement soy protein isolate (SPI), methylcellulose and gluten Vegetarian diet hamburger patti recipe

Table 48. is directed to replaces textured soybean protein concentrate (TSP) and soy protein isolate system with high protein algae piece Into vegetarian diet hamburger patti recipe

Table 49. is directed to replaces soy protein isolate (SPI), methylcellulose and wheat bran with high protein algae homogenate (AH) The recipe for vegetarian diet hamburger patti that matter is made

Table 50. is directed to be separated with high protein algae homogenate replacement textured soybean protein matter concentrate (TSP) with soybean protein The recipe for vegetarian diet hamburger patti that thing is made

In each case, cooking step is:

1. two kinds of textured soybean proteins (as being applicable) are weighed together.

2. adding Part I water (2.5-3 times of TSP weight) in vertical mixer alms bowl, mix 10 minutes.

3. weigh soybean protein concentrate, methylcellulose, gluten and microalgae biomass and be dry mixed.

4. add dry ingredients into vertical mixer.Plus remaining water and mix 5-10 minutes.

5.5. salt and flavor enhancement are weighed.Weigh oil.Added in blender and mix 5 minutes.

6. using mould formation patti (each patti 65-75g), cover and freeze.

In algal biomass (algae piece and algae homogenate) replaces TSP sample, patti is very viscous, does not comparatively have when cooking Structure.Texture is solid when the patti of other adhesive (such as oat, the oat bran and coarse rice powder) productions of addition is cooked.By algae piece The patti for replacing the recipe production of soy protein isolate is more soft than compareing, and more sticky, texture is worse.Contain replacement soybean The patti of the algae homogenate of Protein Separation thing has the robustness and texture for being comparable to control.Generally speaking, replaced with algae homogenate Change most successful in the patti recipe that vegetarian diet hamburger patti that soy protein isolate is made is test and production, be comparable to The patti of vegetarian diet control, but there is the dietary fiber more than almost twice.

Protein rod

Following instance compares conventional protein rod, wherein being replaced with high protein algal biomass (algae piece or algae homogenate (AH)) Change conventional protein sources (soy protein isolate (SPI) and milk protein concentrate (MPC)).

The recipe of the conventional protein rod of table 51.

Table 52. is directed to the recipe that the protein rod that SPI and MPC is made is replaced with high protein algae piece

Table 53., which is directed to, is homogenized the recipe that (AH) replaces the protein rod that SPI and MPC is made with high protein algae

In each case, cooking step is:

1. all slurry compositions of blending.

2. being heated to 190 °F on furnace roof and being kept for 10 minutes in the case of with lid, stir occasionally.

3. leave heating 10 minutes.It is cooled to about 140 °F.

4. merge with dry ingredients.

5. being divided into many parts of sections, shelve overnight.

6. being cut into rod, compound coating is coated as desired and is packed.

Generally speaking, the protein rod being made of the homogenate of high protein algae shows somewhat more preferable than the protein rod being made of algae piece Bonding.Moreover, these compositions are combined together into corn syrup with what algae was homogenized that the protein rod that is made needs minimum.With it is normal The protein rod of rule is compared, and it is more successful composition that the protein rod being made is homogenized with high protein algae：For the egg of similar quantity For white matter and fat, it contains the dietary fiber more than about three times.

Chocolate nutritious drink (meals sub)

Following instance compares conventional chocolate flavouring nutritious drink, wherein being homogenized with high protein algae piece or high protein algae (AH) the chocolate nutritious drink being made replaces conventional protein sources (soy protein isolate (SPI) and milk protein concentrate (MPC))。

Table 54. is directed to the recipe of conventional chocolate nutritious drink

Table 55. is directed to the food that SPI, the chocolate nutritious drink that maltodextrin and milk protein isolate are made are replaced with algae piece Spectrum

Table 56. is directed to is homogenized the chocolate that (AH) replaces SPI, maltodextrin and milk protein isolate are made with high protein algae The recipe of nutritious drink

Compared with the beverage of conventional recipe, the nutritious drink that the homogenate of high protein algae is produced is thicker in vivo.High protein algae piece The nutritious drink of generation is diluter than control beverage.Generally speaking, containing high protein algae be homogenized beverage in this applications more into Work(, produce the thick nutritious drink with high opacity.The nutritious drink being made is homogenized with algae in sugar, fat and protein Conventional beverage is comparable in terms of level, simultaneously containing fiber almost more than ten times.

Example 19

Genotyping with identify other be suitable as the microalgae of food strain

The Genotyping of algae

The following isolated genes group DNA from algal biomass.In 14,000 × g, centrifuge cell is (about from liquid culture 200mg) 5 minutes.Then cell is resuspended in sterile distilled water, 14,000 × g is centrifuged 5 minutes, abandoning supernatant.Will A diameter of about mm single bead is added in biomass, and pipe is placed at least 15 minutes at -80 DEG C.Sample is removed, plus Enter 150 μ l grinding buffer solutions (1% flesh aminoacyl, 0.25M sucrose, 50mM NaCl, 20mM EDTA, 100mM Tris-HCl, pH 8.0, RNaseA 0.5ug/ul).Precipitation settling flux is made by of short duration vortex, 40ul 5M NaCl are then added.Sample is of short duration It is vortexed, then adds 66 μ l 5%CTAB (cetyl trimethylammonium bromide), and the of short duration vortex of last time.Next by sample Product are incubated 10 minutes at 65 DEG C, afterwards centrifuge them 10 minutes in 14,000x g.Supernatant is transferred in new pipe, with 300 μ l phenol:Chloroform:Isoamyl alcohol 12:12:1 extracts once, is then centrifuged 5 minutes in 14,000x g.Gained aqueous phase is transferred to and contained Have in the new pipe of isopropanol (about 190 μ l) of 0.7 volume, mixed by being inverted, is incubated 30 minutes at room temperature or at 4 DEG C It is lower to be incubated overnight.DNA is reclaimed by being centrifuged 10 minutes in 14,000x g.Then the precipitation generated is washed with 70% ethanol Wash twice, then finally washed once with 100% ethanol.Precipitation is air-dried at room temperature 20 to 30 minutes, be then resuspended in In 50 μ l 10mM TrisCl, 1mM EDTA (pH 8.0).

By the total algae DNA of 5 μ l prepared as described above in 10mM Tris (pH 8.0) with 1:50 dilutions.Following setting PCR Reaction, final volume is 20 μ l.10 μ l 2x iProof HF main mixtures (BIO-RAD) are added to 0.4 μ l primers SZ02613 (5 '-TGTTGAAGAATGAGCCGGCGAC-3 ' (the SEQ ID NO of 10mM stock concentrations:24) in).This primer sequence exists Position in Gen Bank accession number L43357 is 567-588, and highly conserved in higher plant and algite genome. Then 0.4 μ l primers SZ02615 (5 '-CAGTGAGCTATTACGCACTC-3 ' (the SEQ ID NO of 10mM stock concentrations are added: 25)).The primer sequence and the position 1112-1093 complementations in Gen Bank accession number L43357, and in higher plant and algae It is highly conserved in plastid genome.Then, the 5 diluted STb genes of μ l and 3.2 μ l dH are added₂O.Following operation PCR reactions：98 DEG C, 45 "；98 DEG C, 8 "；53 DEG C, 12 "；72 DEG C, 20 ", 35 circulations are carried out, is then run 1 minute at 72 DEG C, is maintained at 25 DEG C. For purified pcr product, 20 μ l 10mM Tris (pH 8.0) are added in each reactant, then with 40 μ l phenol:Chlorine It is imitative:Isoamyl alcohol 12:12:1 extracts, and 14,000 × g is vortexed and persistently centrifuged 5 minutes.PCR reactants are applied to S-400 posts (GE Healthcare), 3,000 × g is centrifuged 2 minutes.Then the PCR primer TOPO of purifying is cloned into PCR8/GW/TOPO, Positive colony is selected to be used for LB/Spec plates.The DNA of purifying is entered in the two directions using M13 forward and reverses primer Row sequencing.Sequence alignment and unrooted tree are produced using Geneious DNA analyses software.From 1-23 plants of (institutes in example 1, table 1 Specify) sequence in appended sequence table with SEQ ID NOs:1-23 is listed.

To the 23S rRNA of 9 strains from Chlorella protothecoides genomic DNA analysis

By from Chlorella protothecoides 8 strains (UTEX 25, UTEX 249, UTEX 250, UTEX 256, UTEX 264, UTEX 411, SAG 21110d, CCAP 211/17 and CCAP211/8d) genomic DNA separation, side in accordance with the above Method carries out 23S rRNA genomic DNA analysis.In addition to UTEX 25, all plants of sequence of the Chlorella protothecoides tested It is identical.The sequence of all 8 strains is in appended sequence table with SEQ ID NO:26 and NO:27 list.

The Genotyping analysis of the chlorella sample of purchase

Genotyping is carried out to the chlorella sample commercially available from following 3 using the above method：Chlorella vulgaris (Chlorella regularis) (New Chapter, 390mg/ soft capsule), Whole Foods Broken Cell Wall (NutriBiotic, 500mg/ are pressed by Chlorella (whole blood, 500mg/ compressed tablets) and NutriBiotic CGF Chlorella Film-making).About 200mg every kind of commercially available chlorella sample is resuspended in sterile distilled water for genomic DNA point From.

Gained PCR primer is separated and is cloned into carrier, is sequenced using M13 forward and reverses primer.Searched using BLAST Rope compares sequence with known array.

The comparison of 23s rRNA DNA sequence dnas, which discloses 2 kinds in the chlorella sample commercially available from 3 kinds, has matching existing River mouth sheath silk algae (Lyngbya aestuarii) DNA sequence dna (Whole Foods Broken Wall Chlorella and NutriBiotic CGF).River mouth sheath silk algae is the cyanobacteria (cynobacteria) of ocean species.These results show some Commercially available chlorella contains the species of other contaminating microorganisms, including from being known to toxigenic category (such as sheath Ulothrix) Biology (see, for example, Teneva etc., Environmental Toxicology, 18 (1) 1, the 9-20 pages (2003)； Matthew etc., J Nat Prod., 71 (6)：The 1113-6 pages (2008)；With Carmichael etc., Appl Environ Microbiol, 63 (8)：Page 3104-3110 (1997)).

Example 20

It is suitable as the color change body of the microalgae biomass of food

Produce the mutagenesis of color change body

Method and conditioned growth Chlorella protothecoides (UTEX 250) according to example 1.Use N- methyl-N '-nitre Base-N- nitroguanidines (NTG) carry out mutagenesis to algae strain.Algae culture is set to be subjected to mutagens (NTG), then in 2.0% grape Separated and selected again by number wheel on sugared agar plate.These colonies are screened for color change body.It is former when heterotrophic growth Shell chlorella (wild type) shows gold.Color development produced by being screened on agar plate goes out one plant of white.This face Color mutant is named as 33-55 and (is deposited in American Type culture on October 13rd, 2009 according to budapest treaty to protect Tibetan center (10801Boulevard universities, Manassas, VA 20110-2209), Patent Deposit number is PTA-10397).Also divide Another colony is separated out, it is stable to separate this mutation of confirmation again by three-wheel.This mutant color is on agar plate Be revealed as it is light yellow, be named as 25-32 (according to budapest treaty be deposited on October 13rd, 2009 U.S. typical case culture Thing collection (10801Boulevard universities, Manassas, VA 20110-2209), Patent Deposit number is PTA-10396).

Chlorella protothecoides 33-55 lipodogramme

Method and conditioned growth Chlorella protothecoides 33-55 and parent's Chlorella protothecoides (UTEX according to example 1 250).Determine the lipid percentage of two kinds of strains (with dry cell weight basis)：Chlorella protothecoides 33-55 is 68% lipid, parent plant For 62% lipid.Determine the lipodogramme of two kinds of strains, (being expressed as area %) as follows：Chlorella protothecoides 33-55, C14:0 (0.81)；C16:0(10.35)；C16:1(0.20)；C18:0(4.09)；C18:1(72.16)；C18:2(10.60)；C18:3 (0.10)；And other (1.69)；For parent plant, C14:0(0.77)；C16:0(9.67)；C16:1(0.22)；C18:0 (4.73)；C18:1(71.45)；C18:2(10.99)；C18:3(0.14)；And other (2.05).

Example 21

Cellulosic material for cultivating the microalgae biomass for being suitable as food

In order to assess whether Chlorella protothecoides (UTEX 250) can utilize non-food carbon source, it is prepared for being used as protoconch bead The cellulosic material (pop corn stalk) of the carbon source of the Heterotrophic culture of algae, the Chlorella protothecoides are suitable for use in previous examples In described any food applications.

Wet pop corn stalk material is by National Renewable Energy laboratory (National Renewable Energy Laboratory) (Golden, CO) pass through the boiling and obtained by making in 1.4% sulfuric acid solution by maize straw material De-watering of slurries and prepare.Using Mettler Toledo moisture analysers, it is 24% to determine the dry solid in wet corn stalk.Will The wet samples of 100g are resuspended in deionized water, and it is 420ml to make final volume, and pH then is adjusted into 4.8 using 10N NaOH.Plus Enter Celluclast^TM(Novozymes) (cellulase) makes ultimate density be 4%, and gained slurry is shaken into incubation at 50 DEG C 72 hours.Then the pH of the material is adjusted to 7.5 (insignificant Volume Changes) with NaOH, filtered by 0.22 μm of filter Sterilizing, and stored at -20 DEG C.As described below, keeping sample with using the kit based on hexokinase from Sigma come Determine concentration of glucose.

Concentration of glucose is determined using Sigma glucose assays reagents #G3293.The sample handled as described above is diluted 400 Times, 40 μ l are added in reactant.Corn stalk fiber element preparation is determined the glucose containing about 23g/L.

After carrying out ferment treatment to cellulose and being saccharified into glucose, xylose and other monose, the training described in example 1 is used The material for supporting the above-mentioned preparation of base assessment is used as the raw material grown for Chlorella protothecoides (UTEX250).Pair mixed with pure glucose The cellulose sugars of various concentrations tested (0,12.5,25,50 and 100% cellulose sugar).At 28 DEG C and shake Under (300rpm), in dark place by cell incubation in the cellulose sugar of various concentrations.By in UV spectrophotometers in 750nm Place measures absorbance to evaluate growth.The Chlorella protothecoides culture Celluclast systems grown on maize straw material It is standby, including wherein 100% fermentable sugars is the culture medium condition of cellulose origin.Also use the sweet tea handled with Accellerase Puree carries out similar experiment as cellulosic material.It is similar with the result obtained with maize straw material, all Chlorella protothecoides Culture can be used as carbon source by the use of the sugar of cellulose origin.

PCT Patent application No.:For all purposes, it is entitled by what is submitted on January 19, in 2007 “Nutraceutical Compositions from Microalgae and Related Methods of Production And Administration " PCT Patent Application PCT/US2007/001319 full text is combined hereby.For institute Purposefully, entitled " the Nutraceutical Compositions from that will be submitted on January 19th, 2007 Microalgae and Related Methods of Production and Administration " PCT Patent Application Number PCT/US2007/001653 full text is combined hereby.For all purposes, the title on January 19th, 2007 submitted For " Microalgae-Derived Composition for Improving Health and Appearance of Skin " PCT Patent Application PCT/US2007/001653 full text is combined hereby.For all purposes, will be in 2008 Submit on April 8, entitled " the Fractionation of Oil-Bearing Microbial Biomass " U.S. faces When number of patent application 61/043,620 and entitled " the Direct Chemical that are submitted on April 9th, 2008 Modification of Microbial Biomass " U.S. Provisional Patent Application No. 61/043,620, and in 2010 Entitled " the Methods of Microbial Oil Extraction and Separation " PCT submitted April 14 Number of patent application _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ (attorney docket 026172-004910PC) be each combined with its full text.

No matter previously whether clearly combining, all bibliography cited herein (including patent, patent application and publication Thing) it is combined by reference with its full text.The purpose for being hereby incorporated the publication mentioned is description and openly may be used Reagent, method and the concept being employed in conjunction with the invention.It is in no way to be construed as recognizing that these bibliography are in this institute herein The prior art for the invention stated.

Although invention has been described for combined specific embodiments of the present invention, it should be understood that can To its further modification.Although that the application is intended to generally to follow the principle of the invention including to be not belonging to institute of the invention public Open content category but belong to known technology of the art or conventional technological means and can apply to above carry Any change, purposes or reorganization in the essential feature gone out.

Claims

1. it is a kind of including dry food ingredients composition egg products and microalgae powder, suitable for human consumption, wherein microalgae powder It is the dry homogenate of the microalgae biomass of pasteurization, the microalgae biomass is thin for the Chlorella protothecoides of the dissolving of powder type Born of the same parents, and the microalgae biomass includes triglyceride oil by dry weight between 45% and 65%, wherein the food ingredients group Compound comprise mean for adding liquid with optional other comestible compositions prepare the food of conventional food product into Point, and further wherein described microalgae powder is not green, due to Heterotrophic culture in the dark with the leaf less than 200ppm Chlorophyll contents, and wherein described triglyceride oil is having less than 5% docosahexaenoic acid.

2. a kind of food ingredients composition by combining formed by egg products and microalgae powder, suitable for human consumption, wherein institute The homogenate of drying for the microalgae biomass that microalgae powder is pasteurization is stated, the microalgae biomass is small for the protoconch of the dissolving of powder type Ball frustule, and the microalgae biomass includes triglyceride oil by dry weight between 45% and 65%, wherein the food Component composition comprises mean for adding liquid and optional other comestible compositions to prepare conventional food product Food ingredients, and further wherein described microalgae powder is not green, due to Heterotrophic culture in the dark with being less than 200ppm chlorophyll content, and wherein described triglyceride oil is having less than 5% docosahexaenoic acid.

3. a kind of method for preparing conventional food product, the food article includes combination drying egg products and microalgae powder, wherein The microalgae powder is the homogenate of drying of the microalgae biomass of pasteurization, and the microalgae biomass is small for the protoconch of the dissolving of powder type Ball frustule, and the microalgae biomass includes triglyceride oil by dry weight between 45% and 65%, and further Wherein described algae powder is not green, due to Heterotrophic culture in the dark with the chlorophyll content less than 200ppm, and its Described in oil having less than 5% docosahexaenoic acid.

4. it is a kind of by combining the other comestible compositions of egg products and the slurry and at least one of microalgae powder or microalgae powder simultaneously And conventional food composition formed by heating, the wherein microalgae powder are that the microalgae biomass of pasteurization dries homogenate, The microalgae biomass is the Chlorella protothecoides cell of the dissolving of powder type, and the microalgae biomass includes by dry weight 45% And the triglyceride oil between 65%, and be not green, it is green with the leaf less than 200ppm due to Heterotrophic culture in the dark Cellulose content, and wherein described triglyceride oil is having less than 5% docosahexaenoic acid.

5. a kind of food ingredients composition including egg products and microalgae powder, the wherein microalgae powder are that the microalgae of pasteurization is biological Matter dries homogenate, and the microalgae biomass is the Chlorella protothecoides cell of the dissolving of powder type, and the microalgae biomass bag The triglyceride oil by dry weight between 45% and 65% is included, and wherein 60%-75% triglyceride oil is glyceride shape The 18 of formula:1 lipid, wherein the food ingredients composition comprise mean for addition edible liquid and it is optional other Comestible composition prepare the food ingredients of conventional food product, wherein the microalgae powder is not green, due in the dark Heterotrophic culture and with the chlorophyll content less than 200ppm, and wherein described triglyceride oil is having less than the 20 of 5% Two carbon acids.

6. a kind of microalgae biomass by including triglyceride oil by dry weight between 45% and 65% and it is at least one its The conventional food product of seitan is free of formed by the combination of his powder without seitan or the cereal products without seitan, wherein Microalgae biomass is microalgae powder, and the microalgae powder is the Chlorella protothecoides cell of the dissolving of powder type, and the microalgae biomass is not It is green, due to Heterotrophic culture in the dark with the chlorophyll content less than 200ppm, and wherein described triglycerides Oil is having less than 5% docosahexaenoic acid.

7. a kind of compound powder without seitan, it includes a kind of microalgae powder and at least one others in addition to microalgae powder not Powder containing seitan, the wherein microalgae powder dry homogenate, the wherein microalgae for the Chlorella protothecoides cell of the dissolving of pasteurization Powder is powder type, 22 carbon six containing triglyceride oil by dry weight between 45% and 65% and less than 5% Olefin(e) acid, is not green, and due to Heterotrophic culture in the dark with the chlorophyll content less than 200ppm.

8. formed by a kind of mixture by bakeing microalgae powder, edible liquid and at least one other comestible compositions Conventional food article, the wherein microalgae powder are that drying for the microalgae biomass of pasteurization is homogenized, and the microalgae biomass is The Chlorella protothecoides cell of the dissolving of powder type, and the microalgae biomass is with sweet between 45% and 65% by dry weight The triglycerides oil content of oily three ester oils, having less than 5% docosahexaenoic acid, and it is small due to Heterotrophic culture in the dark In 200ppm chlorophyll content.

9. a kind of method for manufacturing conventional baking goodses, including:

The microalgae biomass of dry, pasteurization Chlorella protothecoides, edible liquid and at least one others are edible With into subassembly and the mixture is bakeed, wherein the microalgae biomass is not green, and due to Heterotrophic culture in the dark and With the chlorophyll content less than 200ppm, and the triglyceride oil between 45% and 65% by dry weight, two less than 5% Dodecahexaene acid, and wherein the microalgae biomass is the full cell sheet or full cell powder form of dissolving.

10. a kind of conventional beverage of microalgae biomass and edible liquid including pasteurization, the wherein microalgae biomass It is not green, comprising the chlorophyll content due to Heterotrophic culture in the dark less than 200ppm, by dry weight 45% and 65% Between triglyceride oil, having less than 5% docosahexaenoic acid, and be dissolving Chlorella protothecoides cell.

11. the method for conventional beverage is manufactured a kind of, including microalgae biomass is combined with edible liquid, wherein the microalgae is given birth to Material is not green, comprising the chlorophyll content due to Heterotrophic culture in the dark less than 200ppm, by dry weight 45% He Triglyceride oil between 65%, having less than 5% docosahexaenoic acid, and is the Chlorella protothecoides cell of dissolving.

12. a kind of food article of conventional fermentation, including：

A. microalgae biomass dry, pasteurization, the microalgae biomass is not green, and comprising due to different in the dark Support and cultivate and the chlorophyll content less than 200ppm, by dry weight the triglyceride oil between 45% and 65%, having less than 5% Docosahexaenoic acid, and be dissolving Chlorella protothecoides cell；

B. a kind of edible liquid；And

C. it is a kind of suitable for the microorganism of the work used in the food article of conventional fermentation.

13. a kind of microalgae powder, it is that drying for the microalgae biomass of pasteurization is homogenized, and the microalgae biomass is powder type Dissolving Chlorella protothecoides cell, wherein the microalgae biomass be not it is green, with due to Heterotrophic culture in the dark and it is small In 200ppm chlorophyll content, and containing triglyceride oil by dry weight between 45% and 65%, having less than the two of 5% Dodecahexaene acid, and processed under the conditions of Good Manufacture Practice (GMP), wherein the microalgae biomass is derived from spreading out from it A kind of algae with the reduced colour mutant of pigment formation is compared in raw strain, and wherein microalgae powder is used to prepare conventional food system Product.

14. a kind of conventional food or food ingredients composition, the microalgae that it contains at least pasteurization of 10% weight are biological The homogenate of matter, the microalgae biomass is not green, and the microalgae biomass is the Chlorella protothecoides cell of dissolving, and containing by dry Triglyceride oil between restatement 45% and 65%, having less than 5% docosahexaenoic acid, and due to heterotrophism in the dark Cultivate and the chlorophyll content less than 200ppm.

15. by slurry formed by scattered microalgae powder, wherein the microalgae powder is the microalgae biomass of dry pasteurization, And be not green, the microalgae biomass is the Chlorella protothecoides cell of dissolving, and the microalgae biomass is included less than 200ppm Chlorophyll content and the by dry weight triglyceride oil between 45% and 65%, having less than 5% docosahexaenoic acid, its In the microalgae powder constitute by weight the 10% to 50% of the slurry, wherein the slurry be used for prepare conventional food product.