WO2020125742A1 - 一种混合真菌分段式固态发酵制备功能性油脂的方法 - Google Patents

一种混合真菌分段式固态发酵制备功能性油脂的方法 Download PDF

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WO2020125742A1
WO2020125742A1 PCT/CN2019/126885 CN2019126885W WO2020125742A1 WO 2020125742 A1 WO2020125742 A1 WO 2020125742A1 CN 2019126885 W CN2019126885 W CN 2019126885W WO 2020125742 A1 WO2020125742 A1 WO 2020125742A1
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fermentation
temperature
extraction
mixed
powder
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French (fr)
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刘元法
李进伟
翟颖红
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Jiangnan University
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    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
    • C11B1/00—Production of fats or fatty oils from raw materials
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
    • C11B1/00—Production of fats or fatty oils from raw materials
    • C11B1/02—Pretreatment
    • C11B1/04—Pretreatment of vegetable raw material
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23B—PRESERVATION OF FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES; CHEMICAL RIPENING OF FRUIT OR VEGETABLES
    • A23B20/00—Preservation of edible oils or fats
    • A23B20/30—Preservation of other edible oils or fats, e.g. shortenings or cooking oils
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23D—EDIBLE OILS OR FATS, e.g. MARGARINES, SHORTENINGS OR COOKING OILS
    • A23D9/00—Other edible oils or fats, e.g. shortenings or cooking oils
    • A23D9/007—Other edible oils or fats, e.g. shortenings or cooking oils characterised by ingredients other than fatty acid triglycerides
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23D—EDIBLE OILS OR FATS, e.g. MARGARINES, SHORTENINGS OR COOKING OILS
    • A23D9/00—Other edible oils or fats, e.g. shortenings or cooking oils
    • A23D9/02—Other edible oils or fats, e.g. shortenings or cooking oils characterised by the production or working-up
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23D—EDIBLE OILS OR FATS, e.g. MARGARINES, SHORTENINGS OR COOKING OILS
    • A23D9/00—Other edible oils or fats, e.g. shortenings or cooking oils
    • A23D9/02—Other edible oils or fats, e.g. shortenings or cooking oils characterised by the production or working-up
    • A23D9/04—Working-up
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
    • A23L11/05—Mashed or comminuted pulses or legumes; Products made therefrom
    • A23L11/07—Soya beans, e.g. oil-extracted soya bean flakes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
    • A23L27/70—Fixation, conservation, or encapsulation of flavouring agents
    • A23L27/72—Encapsulation
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
    • A23L31/10—Yeasts or derivatives thereof
    • A23L31/15—Extracts
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/115—Fatty acids or derivatives thereof; Fats or oils
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/14—Yeasts or derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
    • C11B1/00—Production of fats or fatty oils from raw materials
    • C11B1/02—Pretreatment
    • C11B1/025—Pretreatment by enzymes or microorganisms, living or dead
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
    • C11B1/00—Production of fats or fatty oils from raw materials
    • C11B1/10—Production of fats or fatty oils from raw materials by extracting
    • C11B1/104—Production of fats or fatty oils from raw materials by extracting using super critical gases or vapours
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
    • C11B3/00—Refining fats or fatty oils
    • C11B3/12—Refining fats or fatty oils by distillation
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
    • C11B5/00—Preserving by using additives, e.g. anti-oxidants
    • C11B5/0021—Preserving by using additives, e.g. anti-oxidants containing oxygen
    • C11B5/0035—Phenols; Their halogenated and aminated derivates, their salts, their esters with carboxylic acids
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
    • C11B5/00—Preserving by using additives, e.g. anti-oxidants
    • C11B5/0042—Preserving by using additives, e.g. anti-oxidants containing nitrogen
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids

Definitions

  • the invention relates to a method for preparing functional oil and fat by mixed fungus segmented solid state fermentation, which belongs to the field of edible oil processing.
  • Soybean oil is the most produced vegetable oil in the world.
  • the traditional methods of preparing soybean oil include squeezing and leaching.
  • the squeezing method mainly uses a screw press to squeeze, and the soybean oil obtained by squeezing can retain its original flavor and Nutrient composition, but it has strict requirements on the selection of raw materials, and there is a problem of low oil extraction rate, which will greatly increase its production cost.
  • the principle of the extraction method is to extract soybean oil through organic solvents. The process requires high-temperature deodorization and other refining processes, which will destroy the affected components in the oil and have higher requirements for later refining.
  • the prior art also discloses that the water enzyme method is used for oil extraction. Although the water enzyme method is more environmentally friendly and the raw material utilization rate is higher, it requires higher equipment. In addition, due to the difficulty of oil-water separation, the water enzyme method cannot be used on a large scale. Development and application.
  • Solid-state fermentation refers to the use of natural substrates as carbon sources and energy sources, or inert substrates as solid supports
  • the system is close to the anhydrous fermentation process.
  • the prior art discloses a combination of solid-state fermentation and water enzymatic method or cold-pressing method to ferment soybean or sesame, which can improve the oil extraction rate.
  • the research on the influence of different strains on this process also needs to be further explored and perfected. Finding the appropriate strain is the key to affecting the oil extraction effect and the quality of by-products.
  • Functional oil is a kind of oil that has nutrition and health care functions for the human body, not only has good sensory functions (flavor, appearance), but also has relatively high nutritional functions.
  • Functional oils have become more and more popular.
  • Edible blended oil is an edible oil prepared by blending two or more vegetable fats in proportion.
  • the concept of blended oil in the market is relatively vague.
  • blended oil Before the national standard for blended oil was not promulgated, the fatty acid composition and beneficial concomitant of blended oil were not There are strict restrictions. In order to make profits, many merchants make a fuss about the concept of secondment and oil, and often appear to be low-charged and high-charged.
  • the technical problem to be solved by the present invention is to provide an oil and a preparation method that can satisfy consumers' pursuit of innovative oil and fat products, oil flavor, color and functionality, and at the same time improve the comprehensive utilization rate of resources.
  • the technical solution provided by the present invention is a combination of fungal segmented solid-state fermentation technology and supercritical extraction fractionation technology to prepare different functional oils and fats required for different people.
  • the present invention provides a mixed fungus A segmented solid-state fermentation method for functional oils and fats, the method includes the following steps:
  • Powder preparation After crushing and peeling the soybeans, the billet is rolled, and then expanded using a hydraulic cone plug extruder. The expansion temperature is 150-160°C, the expansion time is 1-3min, and then cooled to 25-35°C to obtain soybeans Powder; peel or hull other oil seeds and pulverize to 50-100 mesh. The crushed powder is autoclaved at 115-121°C for 15-30min and cooled to 25-35°C to obtain other oilseed powder;
  • the fungus is a mold or yeast
  • the mold includes Aspergillus oryzae, Aspergillus niger, and Mortierella isabellina
  • the yeast includes Rhodomyces guttarum, Candida albicans, and Rhodosporidium convolvulum
  • the segment Type fermentation is: when the fungus is a mold, adjust the pH to 3.5-9, the dissolved oxygen content is 30%-40% of the total material volume, first add Aspergillus oryzae to ferment for 1-3 days, then add 0.1-1wt% Glucose, adjust the pH to 3.5-9, the dissolved oxygen content is 30%-40% of the total material volume, then add Aspergillus niger and Mortierella isabellina, and cultivate for 2-6 days; or, when the fungus is yeast, adjust the pH to 5 ⁇ 9, the dissolved oxygen content is 25% ⁇ 35% of the total material volume, first add Candida albicans or Candida albicans and Rhodotorula sphaer
  • the other oilseeds are any one or more of flaxseed, hemp seed, rapeseed, sesame, peanut, corn germ.
  • the mass ratio of the Aspergillus oryzae, Aspergillus niger and Mortierella aureus is 1 to 5:1 to 5:1 to 5.
  • the mass ratio of Rhodopseudomonas guttarum, Candida albicans and Rhodosporidium rubrum is 1 to 5:1 to 5:1 to 5.
  • the extraction pressure is 30-40 MPa
  • the CO 2 flow rate is 15-20 L/h
  • the extraction temperature is 40-50° C.
  • the extraction time is 1.5-2 hours.
  • the reduced-pressure fractionation is divided into first-stage reduced-pressure fractionation and second-stage reduced-pressure fractionation, the pressure of the first-stage reduced-pressure fractionation is 10-20 MPa, and the temperature is 40-50°C; The pressure of the second-stage decompression fractionation is 8-10 MPa, and the temperature is 35-45°C.
  • the present invention innovatively uses mixed fungal fermentation technology to metabolize or convert 2-pentylfuran, which mainly produces beany flavor, through fungus using powder growth and metabolism to produce enzymes, etc., and through other oil seeds such as: Synergy of flaxseed, hemp seed, rapeseed, peanut, etc. to improve the flavor of oils and fats, increase the content of pyrazines, alcohols and other flavoring substances in the oils and fats; Oil production will give the oil a bright red color, which can satisfy consumers' desire for innovative products and further promote the development of new oil products;
  • the method of the present invention improves the fatty acid composition in oils and fats from the source through mixed fungal fermentation to obtain high value-added fatty acids ( ⁇ -linolenic acid, arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid, etc.)
  • the content of unsaturated fatty acids in the oil prepared by the invention can be up to 84%, and the method of the invention can retain the nutritional factors in the oil to the greatest extent, and the retention rate of natural antioxidants can reach 90%;
  • the present invention uses a hydraulic conical plug puffing machine to expand soybeans.
  • This heat treatment method has almost no negative impact on soybean oil nutrients, especially on the quality of protein; and can also inactivate anti-nutritional factors and reduce Microbial pollution, reducing energy consumption during autoclaving; using supercritical extraction technology can achieve green production and reduce the generation of chemical pollution; and the meal after fermentation of mixed fungi will contain a large number of functional peptides, which can be further processed. Increase product added value;
  • the oil extraction rate is also greatly increased, the oil extraction rate can reach up to 95.0%, compared with the unfermented oil extraction rate increased by 40 to 60%, compared to single bacteria
  • the oil extraction rate of various fermentations can be increased by 18 to 27%;
  • the present invention can prepare fats with different flavors and color nutrient fortification, can meet consumers' demand for innovative oils and fats, as well as the pursuit of oil flavors, color and functionality, realize green production, and improve the comprehensive utilization rate of resources .
  • Fig. 1 is a schematic diagram of a method for mixed functional fungi fermentation of functional oil and fat in stages.
  • Oil yield (%) (grease content in raw powder-fat content in powdered meal) / fat content in raw powder ⁇ 100%.
  • Calculation method of unsaturated fatty acid content refer to national standard GB 5009.168-2016.
  • Retention rate of beneficial concomitant in oils and fats 1-(content of beneficial concomitant in oils and fats obtained by cable-type extraction-content of beneficial consumables in supercritical oil extraction by fermentation)/content of beneficial congeners in oils and fats obtained by cable-type extraction ⁇ 100 %.
  • Aspergillus oryzae Inoculate Aspergillus oryzae into potato culture solution and shake culture at 180rpm and 35°C for 2d;
  • Aspergillus niger Inoculate Aspergillus niger in a seed medium containing 3wt% glucose, 0.2wt% magnesium sulfate, and 0.2wt% potassium dihydrogen phosphate, and cultivate at 180rpm and 28°C for 2 days;
  • Mortierella chrysosporium Inoculate Mortierella chrysosporium in the potato culture solution, and add 0.05wt% glucose, 0.005wt% potassium dihydrogen phosphate, 0.015wt% magnesium sulfate, and incubate at 180rpm and 28°C for 2 days.
  • Red Yeast Gummy Inoculate Red Yeast Gummy in a medium containing 4wt% glucose, 0.3wt% yeast powder, 0.2wt% ammonium sulfate, 0.7wt% potassium dihydrogen phosphate, 0.15wt% magnesium sulfate at 180rpm Shake culture for 6 days at °C;
  • Candida albicans was inoculated in a medium containing 2wt% glucose, 0.5wt% yeast powder, 0.8wt% peptone, and cultured with shaking at 180rpm and 30°C for 4 days;
  • Rhodosporidium sphaeroides inoculated with 7wt% glucose, 0.01wt% ammonium sulfate, 0.05wt% yeast powder, 0.04wt% potassium dihydrogen phosphate, 0.15wt% magnesium sulfate medium at 180rpm Shake culture at 30°C for 6 days.
  • the powder After crushing, peeling and rolling the soybeans, the powder is puffed using a hydraulic conical plug puffing machine, the puffing temperature is 150°C, the puffing time is 2min, and it is cooled to 35°C; the flaxseed is crushed to 100 mesh and extinguished by 121°C 15min, cool to 28°C;
  • Puffed soybean powder and autoclaved linseed powder were mixed according to 8:2 (W/W), transferred to a fermentor, the moisture content was adjusted to 20% by weight, glucose was added by 1% by weight, and NaCl added by 0.05% by weight , Adjust the pH to about 5.0, the amount of dissolved oxygen is 30% of the total material volume; add Aspergillus oryzae to the fermentation tank, inoculation amount is 10% (W, the same below), fermentation temperature is 35 °C, fermentation time is 2d; 2d After that, the temperature in the fermentor was reduced to 28°C, supplemented with 0.5wt% glucose, and the Aspergillus niger: Mortierella aureus was added to the fermentor according to 1:1 and the total inoculation amount was 10%. The pH was natural and dissolved oxygen The amount is 40% of the total material volume, the fermentation temperature is 28°C, and the fermentation is 5d;
  • the fermentation product was transferred to an extraction kettle with an extraction pressure of 30 MPa, a CO 2 flow rate of 15 L/h, an extraction temperature of 45° C., and an extraction time of 2 h, followed by reduced pressure fractionation, a first-stage reduced pressure fractionation pressure of 15 MPa, and a temperature of 40° C. ;
  • the secondary decompression fractionation pressure is 8 MPa, the temperature is 35 °C, and the prepared nutrient-enhanced functional oil is preserved by nitrogen filling.
  • the mixed fungi segmented solid-state fermentation method is used to prepare oil and fat.
  • the oil yield is 91.7%
  • Natural antioxidants such as phenol are retained to the greatest extent, the retention rate is 92.0%
  • the relative content of pyrazines, alcohols, phenols and other main flavor substances in the oil and fat is 30.8%, and the flavor of soybean oil and fat is improved.
  • the powder After crushing, peeling and rolling the soybeans, the powder is puffed using a hydraulic conical plug puffing machine, the puffing temperature is 160°C, the puffing time is 2min, and it is cooled to 30°C. Crush the flax seeds to 100 mesh, autoclave at 121°C for 15min, and cool to 30°C;
  • Puffed soybean powder and autoclaved linseed powder were mixed according to 8:2 (W/W), transferred to the fermentor, the moisture content was adjusted to 20wt%, the glucose addition amount was 5wt%, the NaCl addition amount was 0.1%, Adjust the pH to 6.2, the dissolved oxygen content is 30% of the total material volume, and inoculate the Candida albicans: Rhodosporium sphaeroides according to 1:1 (W/W) and the inoculation amount is 10% in the fermentation tank, and ferment.
  • the temperature is 30°C, fermentation for 3d; after 3d, the fermentation temperature is reduced to 28°C, supplemented with 2wt% glucose, the dissolved oxygen content is adjusted to 15% of the total material volume, the pH is adjusted to 6.0, inoculated with 10% red yeast, fermentation Temperature 28°C, fermentation time 4d;
  • the fermentation product was transferred to an extraction kettle with an extraction pressure of 30 MPa, a CO 2 flow rate of 15 L/h, an extraction temperature of 40° C., and an extraction time of 1.5 h.
  • the first-stage decompression fractionation pressure is 10 MPa and the temperature is 40° C.; the second-stage decompression fractionation pressure is 8 MPa and the temperature is 35° C.
  • the functional oils and fats with different flavors and colors are prepared and preserved by nitrogen filling.
  • the mixed fungi segmented solid-state fermentation method was used to prepare oil and fat.
  • the oil extraction rate was 94.6%.
  • the fatty acid composition did not change significantly.
  • the oil flavor was greatly improved with the yeast fermentation reaction.
  • the main flavor substances in the oil, pyrazines and alcohols, The relative content of phenols and other substances is 34.9%.
  • the color of the oil and fat appears reddish, the color of the oil and grease is translucent, and natural antioxidants such as vitamin E and tocopherol, which are beneficial concomitants in the oil and fat, are retained to the greatest extent, and the retention rate is 90.1%.
  • the powder After crushing, peeling and rolling the soybeans, the powder is puffed with a hydraulic conical plug puffing machine, the puffing temperature is 160°C, the puffing time is 1min, and it is cooled to 35°C. Pulverize the hemp seeds to 100 meshes, autoclave at 121°C for 15 min, and cool to 35°C;
  • Puffed soybean powder and autoclaved hemp seed powder were mixed according to 9:1 (W/W), transferred to the fermentor, the moisture content was adjusted to 25wt%, glucose was added 5wt%, NaCl was added 0.05wt%, Adjust the pH to about 5.0, the dissolved oxygen content is 40% of the total material volume; add Aspergillus oryzae to the fermentation tank, the inoculation amount is 8% (W), the fermentation temperature is 35 °C, the fermentation time is 2d; after 2d, the fermentation tank The temperature in the medium was reduced to 28°C, supplemented with 0.1wt% glucose, and the Aspergillus niger: Mortierella aureus was mixed into the fermentor according to 1:3, the total inoculation amount was 8%, the pH was natural, and the dissolved oxygen content was the total material 30% of the volume, fermentation temperature is 28°C, fermentation is 5d;
  • the fermentation product was transferred to an extraction kettle with an extraction pressure of 40 MPa, a CO 2 flow rate of 20 L/h, an extraction temperature of 50°C, and an extraction time of 2 hours.
  • the first-stage decompression fractionation pressure is 20 MPa and the temperature is 50° C.; the second-stage decompression fractionation pressure is 10 MPa and the temperature is 45° C.
  • the prepared good flavor and nutrient-enhanced functional oil and fat are preserved by nitrogen filling.
  • the mixed fungi segmented solid-state fermentation method is used to prepare oil and fat, the oil extraction rate is 89.3%, the content of linoleic acid, linolenic acid and arachidonic acid unsaturated fatty acid is 84.3%, and the beneficial concomitants in oil and fat are vitamin E and tocopherol Natural antioxidants are retained to the greatest extent with a retention rate of 90.6%. Flavor and fermentation of hemp seed oil. The relative content of pyrazines, alcohols and phenols in fermented oils and fats is 48.7%. Soybean oil The "aftertaste" disappeared completely.
  • the powder After crushing, peeling and rolling the soybeans, the powder is puffed using a hydraulic conical plug puffing machine.
  • the puffing temperature is 160 °C
  • the puffing time is 1 min, and it is cooled to 30 °C. Crush the flax seeds to 100 mesh, autoclave at 121°C for 15min, and cool to 30°C;
  • Puffed soybean powder and high-pressure sterilized hemp seed powder were mixed according to 9:1 (W/W), transferred to the fermentor, the moisture content was adjusted to 25wt%, the glucose addition amount was 4wt%, and the NaCl addition amount was 0.2wt %, adjust the pH to 6.2, the dissolved oxygen content is 30% of the total material volume, mix and inoculate Candida albicans: Rhodotorula sphaeroides according to 1:1 (W/W), inoculation amount is 8% in the fermentor , Fermentation temperature is 30°C, fermentation for 3d; after 3d, the fermentation temperature is reduced to 28°C, supplemented with 3wt% glucose, the dissolved oxygen content is adjusted to 10% of the total material volume, the pH is adjusted to 6.0, inoculated with 10% red yeast , Fermentation temperature 28°C, fermentation time 4d;
  • the fermentation product was transferred to an extraction kettle with an extraction pressure of 35 MPa, a CO 2 flow rate of 15 L/h, an extraction temperature of 45°C, and an extraction time of 2 hours.
  • the first-stage decompression fractionation pressure is 20 MPa and the temperature is 40° C.; the second-stage decompression fractionation pressure is 8 MPa and the temperature is 40° C.
  • the nutrient-enhanced functional oils and fats with different flavors and colors are prepared and preserved by nitrogen filling.
  • the mixed fungi segmented solid-state fermentation method is used to prepare oils and fats, the oil extraction rate is 22.1%, the fatty acid composition does not change significantly, the oil flavor is greatly improved with the yeast fermentation reaction, and the main flavor substances in the fermented oils are pyrazines and alcohols , Phenols and other substances relative content of 49.5%. Moreover, the color of the oil and fat appears deep red, and the color of the oil and grease is translucent, and the beneficial concomitant phenols such as vitamin E, tocopherol and phytosterols in the oil and fat are retained to the maximum extent, and the retention rate reaches 91.6%.
  • the powder After crushing, peeling and rolling the soybeans, the powder is puffed using a hydraulic conical plug puffing machine, the puffing temperature is 150°C, the puffing time is 3min, and it is cooled to 35°C. Peanuts are peeled and crushed to 150 mesh, autoclaved at 115°C for 30 min, and cooled to 35°C;
  • Puffed soybean powder and high-pressure sterilized peanut powder were mixed according to 8:2 (W/W), transferred to a fermentor, the moisture content was adjusted to 30wt%, glucose was added 3wt%, NaCl was added 1wt%, the pH was adjusted to About 5.0, the dissolved oxygen content is 40% of the total material volume; Aspergillus oryzae is added to the fermentor, the inoculation amount is 8% (W), the fermentation temperature is 35 °C, and the fermentation time is 2d.
  • the temperature in the fermentor was reduced to 28°C, supplemented with 0.1wt% glucose, and the Aspergillus niger: Mortierella aureus was added to the fermentor according to 1:3 and the total inoculation amount was 8%.
  • the pH was natural and dissolved Oxygen content is 30% of the total material volume, fermentation temperature is 28°C, fermentation is 5d;
  • the fermentation product was transferred to an extraction kettle with an extraction pressure of 30 MPa, a CO 2 flow rate of 20 L/h, an extraction temperature of 45°C, and an extraction time of 2 hours.
  • the first-stage decompression fractionation pressure is 15 MPa and the temperature is 40° C.; the second-stage decompression fractionation pressure is 8 MPa and the temperature is 35° C.
  • the prepared good flavor and nutrient-enhanced functional oil and fat are preserved by nitrogen filling.
  • the mixed fungi segmented solid-state fermentation method is used to prepare oil and fat, the oil extraction rate is 94.9%, the content of linoleic acid, linolenic acid and arachidonic acid unsaturated fatty acid is 80.5%, the beneficial concomitant in the oil and fat are vitamin E and tocopherol , Triterpenes, flavonoids, etc. are retained to the greatest extent, the retention rate reaches 84.5%, the flavor of peanut oil and fermentation together can improve the aftertaste of soybean oil, the main flavor substances in fermented oils are pyrazines and alcohols, phenols and other substances. The content is 20.9%.
  • the powder After crushing, peeling and rolling the soybeans, the powder is puffed by hydraulic conical plug puffing machine, the puffing temperature is 150°C, the puffing time is 3min, and it is cooled to 28°C. Peanuts are peeled and crushed to 150 mesh, autoclaved at 115°C for 30 min, and cooled to 30°C;
  • Puffed soybean powder and autoclaved peanut powder are mixed according to 8:2 (W/W) and transferred to a fermentor, adjusting the moisture content to 25wt%, glucose addition to 5wt%, and NaCl addition to 0.2wt%. Adjust the pH to 6.2, the dissolved oxygen content is 30% of the total material volume, inoculate Candida albicans in the fermentor, the inoculation amount is 8%, the fermentation temperature is 30 °C, fermentation 4d; after 4d, the fermentation temperature is reduced to 28 °C, supplemented with 3wt% glucose, the amount of dissolved oxygen is adjusted to 20% of the total material volume, the pH is adjusted to 5.8, Rhodotorula sphaeroides: Rhodotorula viridis mixed inoculated in the fermentor at a ratio of 1:1, the total inoculated amount is 10%, fermentation temperature 28°C, fermentation time 3d;
  • the fermentation product was transferred to an extraction kettle with an extraction pressure of 30 MPa, a CO 2 flow rate of 15 L/h, an extraction temperature of 40° C., and an extraction time of 1.5 h.
  • the first-stage decompression fractionation pressure is 10 MPa and the temperature is 40°C; the second-stage decompression fractionation pressure is 8 MPa and the temperature is 35°C.
  • Good flavor, color and nutrition-enhanced functional oils and fats are prepared and preserved by nitrogen filling.
  • the mixed fungi stage solid-state fermentation method was used to prepare oil and fat, and the oil extraction rate was 95.0%.
  • the beneficial concomitants in the oil such as vitamin E, tocopherol, triterpenes, and flavonoids, were retained to the greatest extent, and the retention rate reached 83.9%.
  • the flavor and fermentation of peanut oil work together.
  • the relative content of pyrazines, alcohols, phenols and other main flavor substances in fermented oils and fats is 18.7%.
  • the oils appear red and have good transparency.
  • the powder After crushing, peeling and rolling the soybeans, the powder is puffed by hydraulic conical plug puffing machine, the puffing temperature is 160°C, the puffing time is 2min, and it is cooled to 28°C. Peel and crush rapeseed to 50 mesh, autoclave at 121°C for 15min, and cool to 28°C;
  • the fermentation product was transferred to an extraction kettle with an extraction pressure of 30 MPa, a CO 2 flow rate of 20 L/h, an extraction temperature of 45°C, and an extraction time of 2 hours.
  • the first-stage decompression fractionation pressure is 15 MPa and the temperature is 40° C.; the second-stage decompression fractionation pressure is 8 MPa and the temperature is 35° C.
  • the prepared good flavor and nutrient-enhanced functional oil and fat are preserved by nitrogen filling.
  • the mixed fungus staged solid-state fermentation method is used to prepare oil and fat, the oil extraction rate is 92.3%, the content of linoleic acid, linolenic acid and arachidonic acid unsaturated fatty acid is 80.2%, and the beneficial concomitants in oil and fat are vitamin E and tocopherol , Phytosterols, etc. are retained to the greatest extent, with a retention rate of 91.0%.
  • the aftertaste of soybeans and the rapeseed oil flavor of rapeseed oil are effectively improved by fermentation.
  • the relative content of pyrazines, alcohols, phenols and other main flavor substances in fermented oils and fats is 10.8%.
  • the powder After crushing, peeling and rolling the soybeans, the powder is puffed using a hydraulic conical plug puffing machine, the puffing temperature is 160°C, the puffing time is 2min, and it is cooled to 30°C. Peel and crush rapeseed to 50 mesh, autoclave at 121°C for 15 min, and cool to 30°C;
  • Puffed soybean powder and autoclaved rapeseed powder were mixed according to 3:2 (W/W), transferred to the fermentor, the moisture content was adjusted to 30wt%, the glucose addition amount was 5wt%, the NaCl addition amount was 0.2wt% , Adjust the pH to 6.2, the dissolved oxygen content is 30% of the total material volume, inoculate Candida albicans in the fermentor, the inoculation amount is 8%, the fermentation temperature is 30 °C, fermentation 4d; after 4d, the fermentation temperature is reduced to 28°C, supplemented with 3wt% glucose, the dissolved oxygen content is adjusted to 20% of the total material volume, the pH is adjusted to 5.8, Rhodotorula sphaeroides: Rhodotorula viridis mixed and inoculated in the fermentor at a ratio of 1:1, the total inoculated amount 10%, fermentation temperature 28°C, fermentation time 3d;
  • the fermentation product was transferred to an extraction kettle with an extraction pressure of 30 MPa, a CO 2 flow rate of 15 L/h, an extraction temperature of 40° C., and an extraction time of 1.5 h.
  • the first-stage decompression fractionation pressure is 10 MPa and the temperature is 40°C; the second-stage decompression fractionation pressure is 8 MPa and the temperature is 35°C.
  • Good flavor and nutrient-enhanced functional oils and fats are prepared and preserved by nitrogen filling.
  • the mixed fungi segmented solid-state fermentation method is used to prepare oil and fat.
  • the oil extraction rate is 96.1%.
  • the beneficial concomitants in the oil such as vitamin E, tocopherol, and phytosterol, are retained to the greatest extent.
  • the retention rate reaches 84.1%. It can effectively improve the beany smell of soybean oil and rapeseed oil.
  • the relative content of pyrazines, alcohols and phenols in fermented oils and fats is 12.3%. After fermentation, the oils appear dark red.
  • the fermentation process is as follows: Aspergillus oryzae is added to the fermentor, the total inoculation amount is 20% (W), staged fermentation, pH5.0, fermentation temperature is 35 °C, ventilated fermentation, total fermentation The time is 7d. The rest of the steps and conditions are the same as the fermentation conditions in Example 1. At this time, the oil yield is 74.2%, the unsaturated fatty acid content is 64.5%, and the beneficial concomitants in the oil, such as vitamin E, tocopherol, and phytosterol, are retained. The rate is 76.3%, and the relative content of the main flavor substances pyrazines, alcohols and phenols is 7.3%.
  • the fermentation process is as follows: Aspergillus niger is added to the fermentation tank, the total inoculation amount is 20% (W), staged fermentation, natural pH, fermentation temperature is 28 °C, ventilated fermentation, total The fermentation time is 7d, and the remaining steps and conditions are the same as in Example 1.
  • the oil yield is 73.3%
  • the unsaturated fatty acid content is 62.8%
  • the beneficial concomitants in oils are vitamin E, tocopherol, and phytosterols.
  • the equivalent retention rate is 59.6%
  • the relative content of the main flavoring substances pyrazines, alcohols and phenols is 6.4%.
  • Mortierella isabellina is added to the fermentation tank, the total inoculation amount is 20% (W), the pH is natural, the fermentation temperature is 28°C, and the fermentation is ventilated , The total fermentation time is 7d, the remaining steps and conditions are the same as the fermentation conditions of Example 1, the oil yield at this time is 78.6%, the unsaturated fatty acid content is 68.1%, the beneficial concomitants in oils such as vitamin E, tocopherol, The retention rate of plant sterols and other substances was 70.2%, and the relative content of the main flavor substances, pyrazines, alcohols and phenols, was 8.1%.
  • the oil yield can be significantly increased by about 16% to 25%; in addition, the mixed fungus staged fermentation of the present invention can give full play to the synergistic conversion ability of various strains for fatty acids .
  • the use of multi-bacterial staged solid state fermentation combined with supercritical carbon dioxide extraction method can increase the retention rate of beneficial concomitants in fats and oils, which can be increased by 17 -25%;
  • the method of the present invention can significantly increase the content of pyrazines, phenols, alcohols and acids in the oil and fat, and significantly improve the aftertaste in soybean oil products.
  • Fermentation is carried out by using red yeast glucomannan separately.
  • the fermentation process is: adding red yeast glucomannan to the fermentor, the total inoculation amount is 20% (W), pH 6.2, fermentation temperature is 30 °C, ventilated fermentation, total fermentation time 7d, the remaining steps and conditions are the same as the fermentation conditions of Example 2.
  • the oil yield at this time is 78.5%
  • the retention rate of beneficial concomitants in oils such as vitamin E, tocopherol, phytosterols, etc. is 67.9%
  • the main flavor substance pyrazine The relative content of substances, alcohols and phenols is 14.5%.
  • Candida albicans alone for fermentation, the fermentation process is as follows: Candida albicans is added to the fermentor, the total inoculation amount is 20% (W), pH 6.0, fermentation temperature is 30 °C, ventilation fermentation, total fermentation time 7d, the remaining steps and conditions are the same as the fermentation conditions of Example 2, the oil yield is 80.2% at this time, the retention rate of beneficial concomitants in oils such as vitamin E, tocopherol, phytosterol, etc. is 80.4%. The relative content of azines, alcohols and phenols is 13.9%.
  • Rhodosporidium sphaeroides Fermentation of Rhodosporidium sphaeroides alone is used.
  • the fermentation process is as follows: add Rhodosporidium sphaeroides to the fermentor, the total inoculation amount is 20% (W), pH 6.0, fermentation temperature is 30 °C, ventilated fermentation, fermentation The total time is 7 days. The rest of the steps and conditions are the same as the fermentation conditions in Example 2. At this time, the oil yield is 77.9%, and the retention rate of beneficial concomitants in oils, such as vitamin E, tocopherol, and phytosterol, is 73.8%.
  • the main flavor The relative content of substances such as pyrazines, alcohols and phenols was 13.4%.
  • the method of the present invention can maintain the continuous vitality of the bacteria through the method of multi-strain mixed staged fermentation combined with supercritical carbon dioxide extraction, and can significantly increase the oil extraction rate by about 18-20%, and It can increase the content of vitamin E, sterols, triterpenes and flavonoids to a certain extent, which can be increased by 22 to 33%; and the main flavor substances in oils are pyrazines, alcohols, phenols and acids The content is greatly increased.
  • red sticky yeast Rhodosporidium rubrum yeast for solid-state fermentation, then add Candida albicans for fermentation, the remaining steps and conditions are the same as the fermentation conditions of Example 2, the oil extraction rate at this time is 87.3%, unsaturated fatty acid content It was 70.6%, and the relative content of the main flavor substances pyrazines, alcohols, phenols, etc. was 18.6%; in addition, after changing the fermentation sequence, the color of the fat obtained by fermentation was not as bright as the color of the fat prepared in Example 2.
  • soybean: other oilseeds are 2:3, solid fermentation is performed by the methods of Examples 1 and 2, and the oil extraction rate is 86.3% and 87.5%, respectively.
  • Increased use of other oilseeds will increase raw material costs to a certain extent.
  • linseed oil, hemp oil and peanut oil and other oils have strong flavors themselves. When the content is large, they can mask the aftertaste of soybean oil without fermentation. If the oil is further extracted by fermentation, the cost will be further increased; and flax Oil, hemp oil and rapeseed oil itself contain a large amount of unsaturated fatty acids, and contain flavor-producing pyrazines and other main substances. If the ratio is increased, the content of 2-pentylpyridine in the oil will increase, which is easy to cause unpleasant The smell.

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Abstract

本发明公开了一种混合真菌分段式固态发酵制备功能性油脂的方法，属于食用油加工领域。本发明利用大豆作为主要发酵底物，通过对不同功能营养的需求，搭配其他油籽，利用真菌分段式固态发酵，并结合超临界萃取分馏技术，以达到改善大豆油的回味和产生不同风味的目的，同时提高油脂的营养价值。本发明的真菌分段式固态发酵可以提高提油率，改善油脂脂肪酸组成，增加主要风味物质吡嗪类、醇类和酚类等物质的含量，其风味及色泽等感官功能更加符合消费者对创新产品的需求。发酵后的粕可以进一步加工，生产功能性多肽等副产物，实现可持续绿色发展。

Description

一种混合真菌分段式固态发酵制备功能性油脂的方法 技术领域
本发明涉及一种混合真菌分段式固态发酵制备功能性油脂的方法，属于食用油加工领域。
背景技术
大豆油是世界上产量最多的植物油，传统的制备大豆油的方法有压榨法和浸提法，压榨法主要是利用螺旋榨油机进行压榨，压榨得到的大豆油能够保留其原有的风味和营养成分，但是其对原料的选料要求严格，且存在提油率偏低的问题，会大大的增加其生产成本。浸提法的原理是通过有机溶剂萃取大豆油，其过程需要经过高温脱臭等精炼过程，会破坏油中的影响成分，且对后期的精炼要求较高。现有技术中也公开了利用水酶法进行提油，尽管水酶法更加环保，原料利用率较高，但是其对设备要求较高，此外，由于油水分离困难，导致水酶法无法大规模发展与应用。
随着科学技术的发展，固态发酵以其独特的优点(如无“三废”排放)获得了人们的关注，固态发酵是指利用自然底物做碳源和能源，或是惰性底物做固体支持物，体系接近无水的发酵过程。现有技术中公开了利用固态发酵和水酶法或者冷榨结合的方式相结合的技术，对大豆或者芝麻进行发酵，能够提高提油率，但是，目前针对固态发酵的研究较少，尤其是对于不同菌株对此种工艺的影响研究也有待进一步探索完善，找到适当的菌株是影响提油效果与副产物品质的关键所在。
功能性油脂是一种对人体具有营养保健功能的油脂，不仅具有良好的感官功能(风味、外观)，还具有比较高的营养功能。随着社会的发展和人们生活水平的提高，越来越多的人们追求健康生活，功能性油脂成为了越来越多的人的青睐。市场上为了满足消费者对功能性油脂的需求，衍生出一系列调和油和在一级油里面添加脂溶性维生素、酚类、甾醇类等物质。食用调和油是将两种或两种以上的植物油脂按比例调和配制的食用油，目前市场上调和油概念比较模糊，在调和油国家标准未颁布之前，未对调和油脂肪酸组成及有益伴随物有严格限制。很多商家为了赚取利润，并且假借调和油的概念大做文章，经常出现以低充高、以次充好等现象。
因此，急需寻找一种绿色、高效的生产功能性油脂的方法，以满足市场上对功能性油脂的需求。
发明内容
[技术问题]
本发明要解决的技术问题是提供一种可以满足消费者对创新油脂产品、油脂风味、色泽及功能性的追求的油脂及其制备方法，同时提高资源综合利用率。
为了实现上述目的，本发明提供的技术方案是综合真菌分段式固态发酵技术、超临界萃取分馏技术，制备针对不同人群所需的不同功能性油脂，具体地，本发明提供了一种混合真菌分段式固态发酵功能性油脂的方法，所述方法包括以下步骤：
(1)粉料准备：大豆破碎去皮后，轧坯，之后利用液压锥形塞膨化机进行膨化，膨化温度150～160℃，膨化时间1～3min，再冷却至25～35℃，得到大豆粉料；将其他油料种子剥壳或脱皮，粉碎至50～100目，粉碎后的粉料在115～121℃高压灭菌15～30min，冷却至25～35℃，得到其他油籽粉料；
(2)真菌发酵：将膨化后的大豆粉料与高压灭菌的其他油籽粉料配比为6～9：1～4(W/W)混合放入发酵罐，调节发酵罐中物料的水分含量为20～30wt％，葡萄糖添加量0.5～5wt％，NaCl添加量0.05～0.5wt％，调节pH为3.5～9，接种10～20wt％的真菌菌种，进行分段式发酵，发酵温度25～35℃，发酵时间为3～7d；
(3)超临界CO 2萃取分馏：将发酵产物转入萃取釜，进行超临界CO 2萃取，之后进行减压分馏，即可制备不同风味和色泽的营养强化的油脂，向油脂中充氮保藏；
其中，所述真菌为霉菌或酵母，所述霉菌包括米曲霉、黑曲霉和深黄被孢霉，所述酵母包括胶粘红酵母、白色假丝酵母和圆红冬孢酵母；所述分段式发酵为：当真菌为霉菌时，调节pH为3.5～9，溶氧量为总物料体积的30％～40％，先加入米曲菌进行发酵1～3d，之后补加0.1～1wt％的葡萄糖，调节pH为3.5～9，溶氧量为总物料体积的30％～40％，再添加黑曲霉和深黄被孢霉，培养2～6d；或，当真菌为酵母时，调节pH为5～9，溶氧量为总物料体积的25％～35％，先添加白色假丝酵母或白色假丝酵母和圆红冬孢酵母，发酵2～5d，之后补加1％～3％葡萄糖，调节pH为5～9，溶氧量为总物料体积的10％～20％再添加胶粘红酵母和圆红冬孢酵母，或胶粘红酵母，发酵1～4d。
在本发明的一种实施方式中，所述其他油料种子为亚麻籽、火麻籽、菜籽、芝麻、花生、玉米胚芽中的任一种或几种。
在本发明的一种实施方式中，所述米曲霉、黑曲霉和深黄被孢霉的质量比为1～5：1～5：1～5。
在本发明的一种实施方式中，所述胶粘红酵母、白色假丝酵母和圆红冬孢酵母的质量比为1～5：1～5：1～5。
在本发明的一种实施方式中，所述萃取压力30～40MPa，CO 2流速为15～20L/h，萃取温 度为40～50℃，萃取时间为1.5～2h。
在本发明的一种实施方式中，所述减压分馏中分为一级减压分馏和二级减压分馏，所述一级减压分馏的压力为10～20MPa，温度40～50℃；所述二级减压分馏的压力为8～10MPa，温度为35～45℃。
本发明取得的有益效果：
(1)本发明创新性的利用混合真菌发酵技术，通过真菌利用粉料生长代谢产酶等，将最主要产生豆腥味的2-戊基呋喃代谢或转化，并通过与其他油料种子如：亚麻籽、火麻籽、菜籽、花生等的协同作用来改善油脂风味，增加油脂中吡嗪类、醇类等风味物质的含量；本发明采用的胶粘红酵母、圆红冬孢酵母发酵制油，会赋予油脂明亮的红色，可以满足消费者对创新产品的渴望，更加推动油脂新产品开发；
(2)本发明方法通过混合真菌发酵从源头改善油脂中脂肪酸组成，获取高附加值脂肪酸(γ-亚麻酸、花生四烯酸、二十碳五烯酸、二十二碳六烯酸等)，本发明制备得到的油脂中的不饱和脂肪酸含量最高可达84％，且本发明方法能够最大程度保留油脂中的营养因子，天然抗氧化剂的保留率可达90％；
(3)本发明利用液压锥形塞膨化机对大豆进行膨化，这种热处理方式对豆油营养物质几乎没有任何负面影响，特别是对蛋白质的质量无影响；并且还可以灭活抗营养因子，减少微生物污染，降低高压灭菌时的能耗；利用超临界萃取技术可以实现绿色环保生产，减少化学污染的产生；并且混合真菌发酵后的粕中，将含有大量功能性多肽，可以进一步深加工处理，提高产物附加值；
(4)本发明方法通过混合真菌分段式固态发酵，其出油率也大大增加，提油率最高可达95.0％，相比与未经发酵提油率增加40～60％，比单一菌种发酵的提油率可提高18～27％；
(5)本发明能够制备不同风味和色泽营养强化的油脂，能够满足消费者对创新油脂产品的需求，以及对油脂风味、色泽及功能性的追求，实现绿色环保生产，同时提高资源综合利用率。
附图说明
图1是一种混合真菌分段式固态发酵功能性油脂的方法示意图。
具体实施方式
出油率(％)＝(原粉料中油脂含量-粉料粕中油脂含量)/原粉料中油脂含量×100％。
不饱和脂肪酸含量的计算方法：参考国标GB 5009.168-2016。
油脂中的有益伴随物的保留率：1-(索式提取得到油脂中有益伴随物的含量-发酵超临界 提油有益伴随物的含量)/索式提取得到油脂中有益伴随物的含量×100％。
吡嗪类和醇类、酚类物质的相对含量的计算方法：主要风味物质含量/全部风味物质含量×100％。
霉菌活化：
米曲霉：将米曲霉接种于马铃薯培养液中，于180rpm 35℃条件下振荡培养2d；
黑曲霉：将黑曲霉接种于含3wt％葡萄糖、0.2wt％硫酸镁、0.2wt％磷酸二氢钾的种子培养基中，于180rpm 28℃培养2d；
深黄被孢霉：将深黄被孢霉接种于马铃薯培养液中，并加入0.05wt％葡萄糖，0.005wt％的磷酸二氢钾，0.015wt％硫酸镁，于180rpm 28℃培养2d。
酵母菌活化：
胶粘红酵母：将胶粘红酵母接种于含4wt％葡萄糖，0.3wt％酵母粉，0.2wt％硫酸铵，0.7wt％磷酸二氢钾，0.15wt％硫酸镁的培养基中，于180rpm 30℃条件下振荡培养6d；
白色假丝酵母：将白色假丝酵母接种于含2wt％葡萄糖，0.5wt％酵母粉，0.8wt％蛋白胨培养基中，于180rpm 30℃条件下振荡培养4d；
圆红冬孢酵母：将圆红冬孢酵母接种于含7wt％葡萄糖，0.01wt％硫酸铵，0.05wt％酵母粉，0.04wt％磷酸二氢钾，0.15wt％硫酸镁培养基中，于180rpm 30℃条件下振荡培养6d。
实施例1
(1)粉料准备
将大豆破碎、去皮、轧坯后，利用液压锥形塞膨化机对粉料膨化，膨化温度150℃，膨化时间2min，冷却至35℃；将亚麻籽粉碎至100目，利用121℃高压灭菌15min，冷却至28℃；
(2)混合真菌发酵
将膨化后的大豆粉料和高压灭菌亚麻籽粉料按照8：2(W/W)混合，转入发酵罐中，调节水分含量为20wt％，添加葡萄糖1wt％，NaCl添加量0.05wt％，调节pH为5.0左右，溶解氧量为总物料体积的30％；将米曲霉加入发酵罐中，接种量为10％(W，下同)，发酵温度为35℃，发酵时间为2d；2d后将发酵罐中的温度降低至28℃，补充0.5wt％的葡萄糖，将黑曲霉：深黄被孢霉按照1：1，总接种量为10％混合加入发酵罐中，pH自然，溶解氧量为总物料体积的40％，发酵温度为28℃，发酵5d；
(3)超临界CO 2萃取分馏
将发酵产物转入萃取釜中，萃取压力30MPa，CO 2流速为15L/h，萃取温度为45℃，萃 取时间为2h，之后进行减压分馏，一级减压分馏压力为15MPa，温度40℃；二级减压分馏压力为8MPa，温度为35℃，制备得到营养强化功能性油脂通过充氮保藏。
通过该混合真菌分段式固态发酵方法制备油脂，出油率为91.7％，亚油酸、亚麻酸和花生四烯酸等不饱和脂肪酸含量为79.5％，油脂中的有益伴随物维生素E、生育酚等天然抗氧化剂得到最大程度保留，保留率为92.0％，油脂中主要风味物质吡嗪类和醇类、酚类等物质相对含量为30.8％，大豆油脂风味得到改善。
实施例2
(1)粉料准备
将大豆破碎、去皮、轧坯后，利用液压锥形塞膨化机对粉料膨化，膨化温度160℃，膨化时间2min，冷却至30℃。将亚麻籽粉碎至100目，利用121℃高压灭菌15min，冷却至30℃；
(2)混合真菌发酵
将膨化大豆粉料和高压灭菌亚麻籽粉料按照8：2(W/W)混合，转入发酵罐中，调节水分含量为20wt％，葡萄糖添加量为5wt％，NaCl添加量0.1％，调节pH为6.2，溶解氧量为总物料体积的30％，将白色假丝酵母：圆红冬孢酵母按照1：1(W/W)，接种量为10％混合接种于发酵罐中，发酵温度为30℃，发酵3d；3d后将发酵温度降至28℃，补充2wt％葡萄糖，溶解氧量调节为总物料体积的15％，pH为调节为6.0，接种10％的粘红酵母，发酵温度28℃，发酵时间4d；
(3)超临界CO 2萃取分馏
将发酵产物转入萃取釜中，萃取压力30MPa，CO 2流速为15L/h，萃取温度为40℃，萃取时间为1.5h。一级减压分馏压力为10MPa，温度40℃；二级减压分馏压力为8MPa，温度为35℃，制备得到不同风味和色泽的功能性油脂通过充氮保藏。
通过该混合真菌分段式固态发酵方法制备油脂，提油率为94.6％，脂肪酸组成变化不显著，油脂风味随着酵母发酵反应得到极大改善，油脂中主要风味物质吡嗪类和醇类、酚类等物质相对含量为34.9％。且油脂色泽呈现微红，油脂色泽透亮，油脂中的有益伴随物维生素E、生育酚等天然抗氧化剂得到最大程度保留，保留率为90.1％。
实施例3
(1)粉料准备
将大豆破碎、去皮、轧坯后，利用液压锥形塞膨化机对粉料膨化，膨化温度160℃，膨化时间1min，冷却至35℃。将火麻籽破壳粉碎至100目，利用121℃高压灭菌15min，冷 却至35℃；
(2)混合真菌发酵
将膨化大豆粉料和高压灭菌火麻籽粉料按照9：1(W/W)混合，转入发酵罐中，调节水分含量为25wt％，添加葡萄糖5wt％，NaCl添加量0.05wt％，调节pH为5.0左右，溶解氧量为总物料体积的40％；将米曲霉加入发酵罐中，接种量为8％(W)，发酵温度为35℃，发酵时间为2d；2d后将发酵罐中的温度降低至28℃，补充0.1wt％的葡萄糖，将黑曲霉：深黄被孢霉按照1：3，总接种量为8％混合加入发酵罐中，pH自然，溶解氧量为总物料体积的30％，发酵温度为28℃，发酵5d；
(3)超临界CO 2萃取分馏
将发酵产物转入萃取釜中，萃取压力40MPa，CO 2流速为20L/h，萃取温度为50℃，萃取时间为2h。一级减压分馏压力为20MPa，温度50℃；二级减压分馏压力为10MPa，温度为45℃，制备得到良好风味及营养强化功能性油脂通过充氮保藏。
通过该混合真菌分段式固态发酵方法制备油脂，提油率为89.3％，亚油酸、亚麻酸和花生四烯酸不饱和脂肪酸含量为84.3％，油脂中的有益伴随物维生素E、生育酚等天然抗氧化剂得到最大程度保留，保留率为90.6％，火麻籽油的风味以及发酵作用，发酵油脂中主要风味物质吡嗪类和醇类、酚类等物质相对含量为48.7％，大豆油的“回味”完全消失。
实施例4
(1)粉料准备
将大豆破碎、去皮、轧坯后，利用液压锥形塞膨化机对粉料膨化，膨化温度160℃，膨化时间1min，冷却至30℃。将亚麻籽粉碎至100目，利用121℃高压灭菌15min，冷却至30℃；
(2)混合真菌发酵
将膨化大豆粉料和高压灭菌火麻籽粉料按照9：1(W/W)混合，转入发酵罐中，调节水分含量为25wt％，葡萄糖添加量为4wt％，NaCl添加量0.2wt％，调节pH为6.2，溶解氧量为总物料体积的30％，将白色假丝酵母：圆红冬孢酵母按照1：1(W/W)，接种量为8％混合接种于发酵罐中，发酵温度为30℃，发酵3d；3d后将发酵温度降至28℃，补充3wt％葡萄糖，溶解氧量调节为总物料体积的10％，pH为调节为6.0，接种10％的粘红酵母，发酵温度28℃，发酵时间4d；
(3)超临界CO 2萃取分馏
将发酵产物转入萃取釜中，萃取压力35MPa，CO 2流速为15L/h，萃取温度为45℃，萃 取时间为2h。一级减压分馏压力为20MPa，温度40℃；二级减压分馏压力为8MPa，温度为40℃，制备得到不同风味和色泽的营养强化功能性油脂通过充氮保藏。
通过该混合真菌分段式固态发酵方法制备油脂，提油率为22.1％，脂肪酸组成变化不显著，油脂风味随着酵母发酵反应得到极大改善，发酵油脂中主要风味物质吡嗪类和醇类、酚类等物质相对含量为49.5％。且油脂色泽呈现深红，油脂色泽透亮，油脂中的有益伴随物维生素E、生育酚等酚类物质、植物甾醇等得到最大程度保留，保留率达到91.6％。
实施例5
(1)将大豆破碎、去皮、轧坯后，利用液压锥形塞膨化机对粉料膨化，膨化温度150℃，膨化时间3min，冷却至35℃。将花生脱皮粉碎至150目，利用115℃高压灭菌30min，冷却至35℃；
(2)混合真菌发酵：
将膨化大豆粉料和高压灭菌花生粉料按照8：2(W/W)混合，转入发酵罐中，调节水分含量为30wt％，添加葡萄糖3wt％，NaCl添加量1wt％，调节pH为5.0左右，溶解氧量为总物料体积的40％；将米曲霉加入发酵罐中，接种量为8％(W)，发酵温度为35℃，发酵时间为2d。2d后将发酵罐中的温度降低至28℃，补充0.1wt％的葡萄糖，将黑曲霉：深黄被孢霉按照1：3，总接种量为8％混合加入发酵罐中，pH自然，溶解氧量为总物料体积的30％，发酵温度为28℃，发酵5d；
(3)超临界CO 2萃取分馏
将发酵产物转入萃取釜中，萃取压力30MPa，CO 2流速为20L/h，萃取温度为45℃，萃取时间为2h。一级减压分馏压力为15MPa，温度40℃；二级减压分馏压力为8MPa，温度为35℃，制备得到良好风味及营养强化功能性油脂通过充氮保藏。
通过该混合真菌分段式固态发酵方法制备油脂，提油率为94.9％，亚油酸、亚麻酸和花生四烯酸不饱和脂肪酸含量为80.5％，油脂中的有益伴随物维生素E、生育酚、三萜类、黄酮类等得到最大程度保留，保留率达到84.5％，花生油的风味以及发酵共同作用，可以改善豆油回味，发酵油脂中主要风味物质吡嗪类和醇类、酚类等物质相对含量为20.9％。
实施例6
(1)粉料准备
将大豆破碎、去皮、轧坯后，利用液压锥形塞膨化机对粉料膨化，膨化温度150℃，膨化时间3min，冷却至28℃。将花生脱皮粉碎至150目，利用115℃高压灭菌30min，冷却至30℃；
(2)混合真菌发酵：
将膨化大豆粉料和高压灭菌花生粉料按照8：2(W/W)混合，转入发酵罐中，调节水分含量为25wt％，葡萄糖添加量为5wt％，NaCl添加量0.2wt％，调节pH为6.2，溶解氧量为总物料体积的30％，将白色假丝酵母接种于发酵罐中，接种量为8％，发酵温度为30℃，发酵4d；4d后将发酵温度降至28℃，补充3wt％葡萄糖，溶解氧量调节为总物料体积的20％，pH为调节为5.8，圆红冬孢酵母：粘红酵母按1：1比例混合接种于发酵罐中，总接种量为10％，发酵温度28℃，发酵时间3d；
(3)超临界CO 2萃取分馏
将发酵产物转入萃取釜中，萃取压力30MPa，CO 2流速为15L/h，萃取温度为40℃，萃取时间为1.5h。一级减压分馏压力为10MPa，温度40℃；二级减压分馏压力为8MPa，温度为35℃，制备得到良好风味、色泽及营养强化功能性油脂通过充氮保藏。
通过该混合真菌分段式固态发酵方法制备油脂，提油率为95.0％，油脂中的有益伴随物维生素E、生育酚、三萜类、黄酮类等得到最大程度保留，保留率达到83.9％。花生油的风味以及发酵共同作用，发酵油脂中主要风味物质吡嗪类和醇类、酚类等物质相对含量为18.7％，油脂呈现红色，且具有良好的透亮度。
实施例7
(1)粉料准备
将大豆破碎、去皮、轧坯后，利用液压锥形塞膨化机对粉料膨化，膨化温度160℃，膨化时间2min，冷却至28℃。将菜籽脱皮粉碎至50目，利用121℃高压灭菌15min，冷却至28℃；
(2)混合真菌发酵：
将膨化大豆粉料和高压灭菌菜籽粉料按照3：2(W/W)混合，转入发酵罐中，调节水分含量为25wt％，添加葡萄糖1wt％，NaCl添加量0.05wt％，调节pH为5.0左右，溶解氧量为总物料体积的30％；将米曲霉加入发酵罐中，接种量为10％(W)，发酵温度为35℃，发酵时间为2d；2d后将发酵罐中的温度降低至28℃，补充0.5wt％的葡萄糖，将黑曲霉：深黄被孢霉按照1：1，总接种量为10％混合加入发酵罐中，pH自然，溶解氧量为总物料体积的40％，发酵温度为28℃，发酵5d。
(3)超临界CO 2萃取分馏
将发酵产物转入萃取釜中，萃取压力30MPa，CO 2流速为20L/h，萃取温度为45℃，萃取时间为2h。一级减压分馏压力为15MPa，温度40℃；二级减压分馏压力为8MPa，温度 为35℃，制备得到良好风味及营养强化功能性油脂通过充氮保藏。
通过该混合真菌分段式固态发酵方法制备油脂，提油率为92.3％，亚油酸、亚麻酸和花生四烯酸不饱和脂肪酸含量为80.2％，油脂中的有益伴随物维生素E、生育酚、植物甾醇等得到最大程度保留，保留率达到91.0％。大豆的回味和菜籽油的菜油味通过发酵得到有效改善，发酵油脂中主要风味物质吡嗪类和醇类、酚类等物质相对含量为10.8％。
实施例8
(1)粉料准备
将大豆破碎、去皮、轧坯后，利用液压锥形塞膨化机对粉料膨化，膨化温度160℃，膨化时间2min，冷却至30℃。将菜籽脱皮粉碎至50目，利用121℃高压灭菌15min，冷却至30℃；
(2)混合真菌发酵：
将膨化大豆粉料和高压灭菌菜籽粉料按照3：2(W/W)混合，转入发酵罐中，调节水分含量为30wt％，葡萄糖添加量为5wt％，NaCl添加量0.2wt％，调节pH为6.2，溶解氧量为总物料体积的30％，将白色假丝酵母接种于发酵罐中，接种量为8％，发酵温度为30℃，发酵4d；4d后将发酵温度降至28℃，补充3wt％葡萄糖，溶解氧量调节为总物料体积的20％，pH为调节为5.8，圆红冬孢酵母：粘红酵母按1：1比例混合接种于发酵罐中，总接种量为10％，发酵温度28℃，发酵时间3d；
(3)超临界CO 2萃取分馏
将发酵产物转入萃取釜中，萃取压力30MPa，CO 2流速为15L/h，萃取温度为40℃，萃取时间为1.5h。一级减压分馏压力为10MPa，温度40℃；二级减压分馏压力为8MPa，温度为35℃，制备得到良好风味及营养强化功能性油脂通过充氮保藏。
通过该混合真菌分段式固态发酵方法制备油脂，提油率为96.1％，油脂中的有益伴随物维生素E、生育酚、植物甾醇等得到最大程度保留，保留率达到84.1％，通过酵母发酵可以有效改善豆油豆腥味及菜籽油的菜油味，发酵油脂中主要风味物质吡嗪类和醇类、酚类等物质相对含量为12.3％，发酵后油脂呈现暗红色。
对比例1
单独利用米曲霉进行发酵，发酵过程为：将米曲霉加入发酵罐中，总的接种量为20％(W)，分段发酵，pH5.0，发酵温度为35℃，通风发酵，总的发酵时间为7d，其余步骤和条件与实施例1发酵条件相同，此时的出油率为74.2％，不饱和脂肪酸含量为64.5％，油脂中的有益伴随物维生素E、生育酚、植物甾醇等保留率为76.3％，主要风味物质吡嗪类、醇类及酚类 等物质相对含量为7.3％。
同样的，单独利用黑曲霉进行发酵，发酵过程为：将黑曲霉加入发酵罐中，总的接种量为20％(W)，分段发酵，pH自然，发酵温度为28℃，通风发酵，总的发酵时间为7d，其余步骤和条件与实施例1发酵条件相同，此时的出油率为73.3％，不饱和脂肪酸含量为62.8％，油脂中的有益伴随物维生素E、生育酚、植物甾醇等保留率为59.6％，主要风味物质吡嗪类、醇类及酚类等物质相对含量为6.4％。
同样的，单独利用深黄被孢霉进行发酵，发酵过程为：将深黄被孢霉加入发酵罐中，总的接种量为20％(W)，pH自然，发酵温度为28℃，通风发酵，总的发酵时间为7d，其余步骤和条件与实施例1发酵条件相同，此时的出油率为78.6％，不饱和脂肪酸含量为68.1％，油脂中的有益伴随物维生素E、生育酚、植物甾醇等保留率为70.2％，主要风味物质吡嗪类、醇类及酚类等物质相对含量为8.1％。
利用单独的米曲霉、黑曲霉或深黄被孢霉进行发酵，在发酵3～4d后菌种活力大幅度下降，发酵底物得不到充分利用。如果不及时补充碳源和菌种，会降低发酵提油的效率。通过多菌种混合分段式发酵，能够显著的提高出油率，提高了约16％～25％；此外，本发明的混合真菌分段发酵能够充分发挥各菌种的对脂肪酸的协同转化能力，提高油脂中不饱和脂肪酸的含量，提高了约17％～25％；利用多菌种分段式固态发酵结合超临界二氧化碳萃取的方法，可以提高油脂中有益伴随物的保留率，可增加17～25％；此外，本发明方法能够显著的增加油脂中吡嗪类、酚类、醇类及酸类等主要风味物质的含量，明显的改善大豆油产品中的回味。
对比例2
单独利用胶粘红酵母进行发酵，发酵过程为：将胶粘红酵母加入发酵罐中，总的接种量为20％(W)，pH6.2，发酵温度为30℃，通风发酵，发酵总时间为7d，其余步骤和条件与实例2发酵条件相同，此时的出油率为78.5％，油脂中的有益伴随物维生素E、生育酚、植物甾醇等保留率为67.9％，主要风味物质吡嗪类、醇类及酚类等物质相对含量为14.5％。
单独利用白色假丝酵母进行发酵，发酵过程为：将白色假丝酵母加入发酵罐中，总的接种量为20％(W)，pH6.0，发酵温度为30℃，通风发酵，发酵总时间为7d，其余步骤和条件与实施例2发酵条件相同，此时的出油率为80.2％，油脂中的有益伴随物维生素E、生育酚、植物甾醇等保留率为80.4％，主要风味物质吡嗪类、醇类及酚类等物质相对含量为13.9％。
单独利用圆红冬孢酵母进行发酵，发酵过程为：将圆红冬孢酵母加入发酵罐中，总的接种量为20％(W)，pH6.0，发酵温度为30℃，通风发酵，发酵总时间为7d，其余步骤和条件与实施例2发酵条件相同，此时的出油率为77.9％，油脂中的有益伴随物维生素E、生育酚、 植物甾醇等保留率为73.8％，主要风味物质吡嗪类、醇类及酚类等物质相对含量为13.4％。
可见，利用单一酵母发酵无法达到完全发酵，菌种在发酵一段时间会失去活力。相比而言，本发明方法通过多菌种混合分段式发酵结合超临界二氧化碳萃取的方法，能够保持菌种的持续活力，能够明显地提高提油率，提高了约18～20％，并能够在一定程度上提高维生素E、甾醇、三萜及黄酮类等有益伴随物的含量，可增加22～33％；且油脂中主要风味物质吡嗪类、醇类、酚类及酸类的物质含量大大增加。
对比例3
先添加黑曲霉：深黄被孢霉进行固态发酵，再添加米曲霉进行发酵，其余步骤和条件与实施例1发酵条件相同，此时的提油率为86.4％，不饱和脂肪酸含量为72.3％，主要风味物质吡嗪类和醇类、酚类等的相对含量为16.3％。改变三种霉菌的添加顺序，则导致其提油率下降，尤其的，主要风味物质的含量明显降低，仅有实施例1中的50％左右；此外，改变发酵顺序后，在接种米曲霉时，需要重新调整发酵pH值，增加人力成本。
对比例4
先添加粘红酵母：圆红冬孢酵母进行固态发酵，再添加白色假丝酵母进行发酵，其余步骤和条件与实施例2发酵条件相同，此时的提油率为87.3％，不饱和脂肪酸含量为70.6％，主要风味物质吡嗪类和醇类、酚类等的相对含量为18.6％；此外，改变发酵顺序后，发酵得到油脂颜色没有实施例2制备得到的油脂颜色鲜亮。
对比例5
若大豆：其他油籽为2：3时，利用实施例1和2的方法进行固态发酵，此时提油率分别为86.3％和87.5％。其他油籽用量增多，会在一定程度上增加原料成本。此外，亚麻油、火麻油及花生油等油脂自身具有很强的风味，较大的含量时，无需发酵即能够很好掩盖大豆油脂的回味，若再用发酵提油又会进一步增加成本；而且亚麻油、火麻油及菜籽油等本身就含有大量不饱和脂肪酸，且含有产生风味的吡嗪类等主要物质，若增大其比例，会增加油脂中2-戊基吡啶的含量，容易产生不愉快的气味。
虽然本发明已以较佳实施例公开如上，但其并非用以限定本发明，任何熟悉此技术的人，在不脱离本发明的精神和范围内，都可做各种的改动与修饰，因此本发明的保护范围应该以权利要求书所界定的为准。

Claims (18)

  1. 一种混合真菌分段式固态发酵制备功能性油脂的方法，其特征在于，所述方法包括以下步骤：
    (1)粉料准备：大豆破碎去皮后，轧坯，之后利用液压锥形塞膨化机进行膨化，膨化温度150～160℃，膨化时间1～3min，再冷却至25～35℃，得到大豆粉料；将其他油料种子剥壳或脱皮，粉碎至50～100目，粉碎后的粉料在115～121℃高压灭菌15～30min，冷却至25～35℃，得到其他油籽粉料；
    (2)真菌发酵：将膨化后的大豆粉料与高压灭菌的其他油籽粉料按照质量配比为6～9：1～4混合放入发酵罐，调节发酵罐中物料的水分含量为20～30wt％，葡萄糖添加量0.5～5wt％，NaCl添加量0.05～0.5wt％，调节pH为3.5～9，接种10～20wt％的真菌菌种，进行分段式发酵，发酵温度25～35℃，发酵时间为3～7d；
    (3)超临界CO 2萃取分馏：将发酵产物转入萃取釜，进行超临界CO 2萃取，之后进行减压分馏，即可制备不同风味和色泽的营养强化的油脂，向油脂中充氮保藏；
    其中，所述真菌为霉菌或酵母，所述霉菌包括米曲霉、黑曲霉和深黄被孢霉，所述酵母包括胶粘红酵母、白色假丝酵母和圆红冬孢酵母；所述分段式发酵为：当真菌为霉菌时，调节pH为3.5～9，溶氧量为总物料体积的30％～40％，先加入米曲菌进行发酵1～3d，之后补加0.1～1wt％的葡萄糖，调节pH为3.5～9，溶氧量为总物料体积的30％～40％，再添加黑曲霉和深黄被孢霉，培养2～6d；或，当真菌为酵母时，调节pH为5～9，溶氧量为总物料体积的25％～35％，先添加白色假丝酵母，或白色假丝酵母和圆红冬孢酵母，发酵2～5d，之后补加1％～3wt％葡萄糖，调节pH为5～9，溶氧量为总物料体积的10％～20％再添加胶粘红酵母和圆红冬孢酵母，或胶粘红酵母，发酵1～4d。
  2. 根据权利要求1所述的一种混合真菌分段式固态发酵制备功能性油脂的方法，其特征在于，所述其他油料种子为亚麻籽、火麻籽、菜籽、芝麻、花生、玉米胚芽中的任一种或几种。
  3. 根据权利要求1或2所述的一种混合真菌分段式固态发酵制备功能性油脂的方法，其特征在于，所述大豆粉料与其他油籽粉料的质量比为6～9：1～4。
  4. 根据权利要求1～3任一所述的一种混合真菌分段式固态发酵制备功能性油脂的方法，其特征在于，所述米曲霉、黑曲霉和深黄被孢霉的质量比为1～5：1～5：1～5。
  5. 根据权利要求1～4任一所述的一种混合真菌分段式固态发酵制备功能性油脂的方法，其特征在于，所述胶粘红酵母、白色假丝酵母和圆红冬孢酵母的质量比为1～5：1～5：1～5。
  6. 根据权利要求1～5任一所述的一种混合真菌分段式固态发酵制备功能性油脂的方法，所述萃取压力30～40MPa，CO 2流速为15～20L/h，萃取温度为40～50℃，萃取时间为1.5～2h。
  7. 根据权利要求1～6任一所述的一种混合真菌分段式固态发酵制备功能性油脂的方法，其特征在于，所述减压分馏中分为一级减压分馏和二级减压分馏，所述一级减压分馏的压力为10～20MPa，温度40～50℃；所述二级减压分馏的压力为8～10MPa，温度为35～45℃。
  8. 根据权利要求1所述的一种混合真菌分段式固态发酵制备功能性油脂的方法，其特征在于，所述方法包括以下步骤：
    (1)粉料准备：将大豆破碎、去皮、轧坯后，利用液压锥形塞膨化机对粉料膨化，膨化温度150℃，膨化时间2min，冷却至35℃；将亚麻籽粉碎至100目，利用121℃高压灭菌15min，冷却至28℃；
    (2)混合真菌发酵：将膨化后的大豆粉料和高压灭菌亚麻籽粉料按照8：2混合，转入发酵罐中，调节水分含量为20wt％，添加葡萄糖1wt％，NaCl添加量0.05wt％，调节pH为5.0左右，溶解氧量为总物料体积的30％；将米曲霉加入发酵罐中，接种量为10％，发酵温度为35℃，发酵时间为2d；2d后将发酵罐中的温度降低至28℃，补充0.5wt％的葡萄糖，将黑曲霉：深黄被孢霉按照1：1，总接种量为10％混合加入发酵罐中，溶解氧量为总物料体积的40％，发酵温度为28℃，发酵5d；
    (3)超临界CO 2萃取分馏：将发酵产物转入萃取釜中，萃取压力30MPa，CO 2流速为15L/h，萃取温度为45℃，萃取时间为2h，之后进行减压分馏，一级减压分馏压力为15MPa，温度40℃；二级减压分馏压力为8MPa，温度为35℃，制备得到营养强化功能性油脂通过充氮保藏。
  9. 根据权利要求1所述的一种混合真菌分段式固态发酵制备功能性油脂的方法，其特征在于，所述方法包括以下步骤：
    (1)粉料准备：将大豆破碎、去皮、轧坯后，利用液压锥形塞膨化机对粉料膨化，膨化温度160℃，膨化时间2min，冷却至30℃；将亚麻籽粉碎至100目，利用121℃高压灭菌15min，冷却至30℃；
    (2)混合真菌发酵：将膨化大豆粉料和高压灭菌亚麻籽粉料按照8：2混合，转入发酵罐中，调节水分含量为20wt％，葡萄糖添加量为5wt％，NaCl添加量0.1％，调节pH为6.2，溶解氧量为总物料体积的30％，将白色假丝酵母：圆红冬孢酵母按照1：1，接种量为10％混合接种于发酵罐中，发酵温度为30℃，发酵3d；3d后将发酵温度降至28℃，补充2wt％葡萄糖，溶解氧量调节为总物料体积的15％，pH为调节为6.0，接种10％的粘红酵母，发酵温度28℃，发酵时间4d；
    (3)超临界CO 2萃取分馏：将发酵产物转入萃取釜中，萃取压力30MPa，CO 2流速为15L/h，萃取温度为40℃，萃取时间为1.5h；一级减压分馏压力为10MPa，温度40℃；二 级减压分馏压力为8MPa，温度为35℃，制备得到不同风味和色泽的功能性油脂通过充氮保藏。
  10. 根据权利要求1所述的一种混合真菌分段式固态发酵制备功能性油脂的方法，其特征在于，所述方法包括以下步骤：
    (1)粉料准备：将大豆破碎、去皮、轧坯后，利用液压锥形塞膨化机对粉料膨化，膨化温度160℃，膨化时间1min，冷却至35℃；将火麻籽破壳粉碎至100目，利用121℃高压灭菌15min，冷却至35℃；
    (2)混合真菌发酵：将膨化大豆粉料和高压灭菌火麻籽粉料按照9：1混合，转入发酵罐中，调节水分含量为25wt％，添加葡萄糖5wt％，NaCl添加量0.05wt％，调节pH为5.0左右，溶解氧量为总物料体积的40％；将米曲霉加入发酵罐中，接种量为8％，发酵温度为35℃，发酵时间为2d；2d后将发酵罐中的温度降低至28℃，补充0.1wt％的葡萄糖，将黑曲霉：深黄被孢霉按照1：3，总接种量为8％混合加入发酵罐中，溶解氧量为总物料体积的30％，发酵温度为28℃，发酵5d；
    (3)超临界CO 2萃取分馏：将发酵产物转入萃取釜中，萃取压力40MPa，CO 2流速为20L/h，萃取温度为50℃，萃取时间为2h；一级减压分馏压力为20MPa，温度50℃；二级减压分馏压力为10MPa，温度为45℃，制备得到良好风味及营养强化功能性油脂通过充氮保藏。
  11. 根据权利要求1所述的一种混合真菌分段式固态发酵制备功能性油脂的方法，其特征在于，所述方法包括以下步骤：
    (1)粉料准备：将大豆破碎、去皮、轧坯后，利用液压锥形塞膨化机对粉料膨化，膨化温度160℃，膨化时间1min，冷却至30℃；将亚麻籽粉碎至100目，利用121℃高压灭菌15min，冷却至30℃；
    (2)混合真菌发酵：将膨化大豆粉料和高压灭菌火麻籽粉料按照9：1混合，转入发酵罐中，调节水分含量为25wt％，葡萄糖添加量为4wt％，NaCl添加量0.2wt％，调节pH为6.2，溶解氧量为总物料体积的30％，将白色假丝酵母：圆红冬孢酵母按照1：1，接种量为8％混合接种于发酵罐中，发酵温度为30℃，发酵3d；3d后将发酵温度降至28℃，补充3wt％葡萄糖，溶解氧量调节为总物料体积的10％，pH调节为6.0，接种10％的粘红酵母，发酵温度28℃，发酵时间4d；
    (3)超临界CO 2萃取分馏：将发酵产物转入萃取釜中，萃取压力35MPa，CO 2流速为15L/h，萃取温度为45℃，萃取时间为2h；一级减压分馏压力为20MPa，温度40℃；二级减压分馏压力为8MPa，温度为40℃，制备得到不同风味和色泽的营养强化功能性油脂通过 充氮保藏。
  12. 根据权利要求1所述的一种混合真菌分段式固态发酵制备功能性油脂的方法，其特征在于，所述方法包括以下步骤：
    (1)粉料准备：将大豆破碎、去皮、轧坯后，利用液压锥形塞膨化机对粉料膨化，膨化温度150℃，膨化时间3min，冷却至35℃；将花生脱皮粉碎至150目，利用115℃高压灭菌30min，冷却至35℃；
    (2)混合真菌发酵：将膨化大豆粉料和高压灭菌花生粉料按照8：2混合，转入发酵罐中，调节水分含量为30wt％，添加葡萄糖3wt％，NaCl添加量1wt％，调节pH为5.0左右，溶解氧量为总物料体积的40％；将米曲霉加入发酵罐中，接种量为8％，发酵温度为35℃，发酵时间为2d；2d后将发酵罐中的温度降低至28℃，补充0.1wt％的葡萄糖，将黑曲霉：深黄被孢霉按照1：3，总接种量为8％混合加入发酵罐中，溶解氧量为总物料体积的30％，发酵温度为28℃，发酵5d；
    (3)超临界CO 2萃取分馏：将发酵产物转入萃取釜中，萃取压力30MPa，CO 2流速为20L/h，萃取温度为45℃，萃取时间为2h；一级减压分馏压力为15MPa，温度40℃；二级减压分馏压力为8MPa，温度为35℃，制备得到良好风味及营养强化功能性油脂通过充氮保藏。
  13. 根据权利要求1所述的一种混合真菌分段式固态发酵制备功能性油脂的方法，其特征在于，所述方法包括以下步骤：
    (1)粉料准备：将大豆破碎、去皮、轧坯后，利用液压锥形塞膨化机对粉料膨化，膨化温度150℃，膨化时间3min，冷却至28℃；将花生脱皮粉碎至150目，利用115℃高压灭菌30min，冷却至30℃；
    (2)混合真菌发酵：将膨化大豆粉料和高压灭菌花生粉料按照8：2混合，转入发酵罐中，调节水分含量为25wt％，葡萄糖添加量为5wt％，NaCl添加量0.2wt％，调节pH为6.2，溶解氧量为总物料体积的30％，将白色假丝酵母接种于发酵罐中，接种量为8％，发酵温度为30℃，发酵4d；4d后将发酵温度降至28℃，补充3wt％葡萄糖，溶解氧量调节为总物料体积的20％，pH为调节为5.8，圆红冬孢酵母：粘红酵母按1：1比例混合接种于发酵罐中，总接种量为10％，发酵温度28℃，发酵时间3d；
    (3)超临界CO 2萃取分馏：将发酵产物转入萃取釜中，萃取压力30MPa，CO 2流速为15L/h，萃取温度为40℃，萃取时间为1.5h；一级减压分馏压力为10MPa，温度40℃；二级减压分馏压力为8MPa，温度为35℃，制备得到良好风味、色泽及营养强化功能性油脂通过充氮保藏。
  14. 根据权利要求1所述的一种混合真菌分段式固态发酵制备功能性油脂的方法，其特征在于，所述方法包括以下步骤：
    (1)粉料准备：将大豆破碎、去皮、轧坯后，利用液压锥形塞膨化机对粉料膨化，膨化温度160℃，膨化时间2min，冷却至28℃；将菜籽脱皮粉碎至50目，利用121℃高压灭菌15min，冷却至28℃；
    (2)混合真菌发酵：将膨化大豆粉料和高压灭菌菜籽粉料按照3：2混合，转入发酵罐中，调节水分含量为25wt％，添加葡萄糖1wt％，NaCl添加量0.05wt％，调节pH为5.0左右，溶解氧量为总物料体积的30％；将米曲霉加入发酵罐中，接种量为10％，发酵温度为35℃，发酵时间为2d；2d后将发酵罐中的温度降低至28℃，补充0.5wt％的葡萄糖，将黑曲霉：深黄被孢霉按照1：1，总接种量为10％混合加入发酵罐中，溶解氧量为总物料体积的40％，发酵温度为28℃，发酵5d；
    (3)超临界CO 2萃取分馏
    将发酵产物转入萃取釜中，萃取压力30MPa，CO 2流速为20L/h，萃取温度为45℃，萃取时间为2h；一级减压分馏压力为15MPa，温度40℃；二级减压分馏压力为8MPa，温度为35℃，制备得到良好风味及营养强化功能性油脂通过充氮保藏。
  15. 根据权利要求1所述的一种混合真菌分段式固态发酵制备功能性油脂的方法，其特征在于，所述方法包括以下步骤：
    (1)粉料准备：将大豆破碎、去皮、轧坯后，利用液压锥形塞膨化机对粉料膨化，膨化温度160℃，膨化时间2min，冷却至30℃；将菜籽脱皮粉碎至50目，利用121℃高压灭菌15min，冷却至30℃；
    (2)混合真菌发酵：
    将膨化大豆粉料和高压灭菌菜籽粉料按照3：2混合，转入发酵罐中，调节水分含量为30wt％，葡萄糖添加量为5wt％，NaCl添加量0.2wt％，调节pH为6.2，溶解氧量为总物料体积的30％，将白色假丝酵母接种于发酵罐中，接种量为8％，发酵温度为30℃，发酵4d；4d后将发酵温度降至28℃，补充3wt％葡萄糖，溶解氧量调节为总物料体积的20％，pH为调节为5.8，圆红冬孢酵母：粘红酵母按1：1比例混合接种于发酵罐中，总接种量为10％，发酵温度28℃，发酵时间3d；
    (3)超临界CO 2萃取分馏：将发酵产物转入萃取釜中，萃取压力30MPa，CO 2流速为15L/h，萃取温度为40℃，萃取时间为1.5h；一级减压分馏压力为10MPa，温度40℃；二级减压分馏压力为8MPa，温度为35℃，制备得到良好风味及营养强化功能性油脂通过充氮保藏。
  16. 权利要求1～15任一所述的一种混合真菌分段式固态发酵制备功能性油脂的方法制备得到的油脂。
  17. 包含权利要求16所述的油脂的食品或食用油。
  18. 权利要求1～15任一所述的一种混合真菌分段式固态发酵制备功能性油脂的方法在食品加工领域的应用。
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