Classifications

• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING OR TREATMENT THEREOF
  • A23C11/00—Milk substitutes, e.g. coffee whitener compositions
  • A23C11/02—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins
  • A23C11/10—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins
  • A23C11/103—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins containing only proteins from pulses, oilseeds or nuts, e.g. nut milk
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
  • A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
  • A23J1/14—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
  • A23J3/00—Working-up of proteins for foodstuffs
  • A23J3/14—Vegetable proteins
  • A23J3/16—Vegetable proteins from soybean
• • 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
• • 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
  • A23L11/60—Drinks from legumes, e.g. lupine drinks
  • A23L11/65—Soy drinks
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P3/00—Drugs for disorders of the metabolism
  • A61P3/06—Antihyperlipidemics
• • 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

Definitions

• the present invention relates to a fractionated soybean protein material, a processed soybean suitable for the same, and a method for producing them. More specifically, the present invention relates to a fractionation technique for proteins having various characteristics (7S globulins, 11S globulins, lipophilic proteins, etc.) contained in soybean protein.
• Soy protein is widely used for improving the physical properties of foods because of its unique gel-forming ability, and it is also highly nutritious and has been increasingly used as a health food material.
• Soybean storage protein precipitates around pH 4.5, and it is relatively easily divided into an acid-soluble protein fraction mainly composed of soluble components other than the storage protein and an acid-precipitated protein fraction mainly composed of storage protein. Can be divided.
• a product obtained by collecting the acid-precipitating protein fraction is a separated soybean protein, which is currently widely used in the food industry.
• Proteins constituting soy protein are classified into 2S, 7S, US, and 15S globulins based on the sedimentation coefficient by ultracentrifugation analysis.
• 7S globulin and 11S globulin are the main constituent protein components of the globulin fraction.
• ⁇ -conglycinin substantially corresponds to 7S globulin
• glycinin substantially corresponds to 11S globulin.
• Proteins constituting soy protein have different properties in terms of physical properties such as viscosity, coagulability, and surface activity, and nutritional management functions.
• Non-patent Document 1 7S globulin has been reported to reduce blood neutral fat (Non-patent Document 1). 11S globulin is said to dominate the hardness of the tofu gel, which has a high gelling power.
• fractionating soy protein into a fraction rich in these components makes it possible to greatly express the functions specific to each protein in terms of physiological functions and physical properties, and is a characteristic material. May lead to the creation of And this allows protein in the food industry Expansion of application fields can be expected.
• Patent Document 2 Patent Documents 1-7, etc.
• acid-precipitable soy protein contains a mixture of proteins with high affinity with polar lipids that make up cell membranes, protein bodies, and oil bodies. It has been reported in recent years (Non-patent Document 3).
• Non-patent Document 4 As a result of research by the present inventor after receiving strong reports, when sodium sulfate was added to low-denatured defatted soymilk to a concentration of 1M and the pH was adjusted to 4.5 with hydrochloric acid, 7S and It was found that 11S globulin migrated, and that other miscellaneous proteins migrated to the acid-precipitating fraction (Non-patent Document 4).
• the amount of nitrogen in this acid-precipitable fraction accounted for about 30% of the total amount of nitrogen in defatted soymilk, which was surprisingly high.
• Non-Patent Document 5 Non-Patent Document 5
• Proteins contained in the acid-precipitated fraction with a small amount of 7S globulin and 11S globulin are mainly proteins showing 34 kDa, 24 kDa and 18 kDa in molecular weight estimated by SDS-polyacrylamide electrophoresis, lipoxygenase, ⁇ -conglycinin, And many other miscellaneous proteins It is a mixture of qualities. This group of proteins exhibits affinity for polar lipids.
• Non-patent Documents 2, Patent Documents 1 to 7 show that lipid-friendly proteins account for a considerable proportion of acid-precipitated soybean protein. Since no consideration was given, it was found that it was practically impossible to fractionate 7S globulin and 11 S globulin with high purity.
• Non-Patent Document 4 shows a high ionic strength, and many reducing agents are required. Therefore, since desalting and washing are essential steps, it is effective at the experimental level, but unsuitable for industrial processes.
• the present applicant has developed a technique for fractionating into high-purity soybean 7S globulin protein and soybean 11S globulin protein with a low contamination rate of lipophilic protein (Patent Documents 8 and 9).
• This method is industrially superior in that 7S globulin is fractionated with high purity.
• 11S globulin also has room for improvement because it requires complicated operations to reduce the contamination with lipophilic protein and fractionate with high purity.
• Non-patent literature l Okita T et al, J. Nutr. Sci. Vitaminol., 27 (4), 379-388, 1981
• Non-Patent Document 2 Thahn, V.H, and Shibasaki.K., J. Agric. Food Chem., 24, 117, 1976
• Non-Patent Document 3 Herman, Planta, 172, 336-345, 1987
• Non-Patent Document 4 Samoto M et al., Biosci. Biotechnol. Biochem., 58 (11), 2123-2125, 1 994
• Non-patent literature 5 Samoto M et al., Biosci Biotechnol Biochem, 62 (5), 935-940, 1998
• Non-patent literature b T. Nagano, et.al., Relationship between rheological properties and conformational states of 7S globulin from soybeans at acidic pH, Food Hydrocolloids: Structures, Properties, and Functions, Plenum Press, New York, 1994
• Patent Document 1 Japanese Patent Application Laid-Open No. 55-124457
• Patent Document 2 JP-A-48-56843
• Patent Document 3 JP-A-49-31843
• Patent Document 4 JP-A-58-36345
• Patent Document 5 Japanese Patent Laid-Open No. 61-187755
• Patent Document 6 International Publication WO00Z58492
• Patent Document 7 US Pat. No. 6,171,640
• Patent Document 8 International Publication WO02Z28198
• Patent Document 9 International Publication WO2004Z43160
• an object of the present invention is to provide means capable of efficiently and highly fractionating not only 7S globulin but also three protein fractions of 11S globulin and lipophilic protein. . It is another object of the present invention to provide a soy milk and a separated soy protein having a reduced lipophilic protein content. It is also an issue that these methods are processes that can be implemented at the food industry level.
• the present inventors prepared a processed soybean obtained by subjecting a low-denatured soybean containing protein and okara to a specific protein modification treatment, and using this as a raw material. Soy milk can be extracted as a high-purity fraction of soybean protein and 7S globulin, 11S globulin, or lipophilic protein can be efficiently separated simply by a simple fractionation method. It reached.
• the present inventors prepared processed soybeans that had been subjected to denaturation treatment under conditions such that 7S globulin and 11S globulin remained low denatured and only the lipophilic protein was selectively denatured. As a result, it was found that 7S globulin and 11S globulin were mainly extracted, while extraction of lipophilic protein was suppressed and a considerable amount remained on the okara side as an insoluble fraction.
• the pH of the obtained soy milk with low lipophilic protein was adjusted to 7S globulin and 11S globulin. It was found that a high-purity fraction of both globulins could be easily achieved simply by adjusting to a pH range where the solubility difference of Brin was large.
• soy milk and isolated soy protein obtained from processed soybeans in which lipophilic proteins are selectively denatured are superior in flavor compared to those produced by conventional methods.
• processed soybean power obtained by selectively modifying a lipophilic protein is rich in lipophilic protein.
• the method for fractionating soybean protein according to the present invention can be applied to the fractionation of two fractions of 11S globulin and lipophilic protein when 7S globulin-deficient soybean is used. I found it.
• Processed soybeans that contain protein and okara components have a PDI of 40 or more and less than 80, and the lipophilic protein is selectively water-insolubilized among the contained proteins
• soybean power as described in 1 above.
• at least one acid-precipitating soybean protein selected from the group consisting of 7S globulin, 11S globulin and lipophilic protein is concentrated.
• a method for producing fractionated soy protein characterized by collecting fractions;
• a method for producing a soy 11S globulin protein comprising adjusting the soy milk having the processed soybean power described in 1. above to pH 5.2 to 6.4 and collecting an insoluble fraction,
• Soybean 7S globulin protein obtained by fractionating soy milk prepared with processed soybean power as described in 1. above
• soybean power as described in 1. above.
• the prepared soymilk was adjusted to pH 4 to 5.5, heated at 40 to 65 ° C, then heated to pH 5.3 to 5.7, and the resulting insoluble fraction was separated and obtained.
• a method for producing soybean 7S globulin protein characterized in that the water-soluble fraction is adjusted to pH 4-5 and the insoluble fraction is recovered;
• soybean power as described in 1. above is fractionated and contains at least 7% oil extracted with a solvent having a volume ratio of black mouth form to methanol of 2: 1.
• non 7S-11S-acid-precipitated soy protein is characterized in that the processed soybean power as described in 1. above is fractionated and contains at least 7% oil extracted with a solvent having a volume ratio of black mouth form to methanol of 2: 1.
• non 7S-11S-acid-precipitated soy protein is characterized in that the processed soybean power as described in 1. above is fractionated and contains at least 7% oil extracted with a solvent having a volume ratio of black mouth form to methanol of 2: 1.
• non 7S-11S-acid-precipitated soy protein is characterized in that the processed soybean power as described in 1. above is fractionated and contains at least 7% oil extracted with a solvent having a volume ratio of black mouth form to methanol of 2: 1.
• non 7S-11S-acid-precipitated soy protein is characterized in that
• a method for producing a non-7S ⁇ 11S-acid-precipitated soy protein characterized by recovering an insoluble fraction produced by adjusting the pH to 5.3 to 5.7 as described in 14.
• a method for fractionating soy protein comprising the following steps: (1) a step of adding the soy milk to okara by adding water to the processed soybean as described in 1. above, and (2) adjusting the soy milk to pH 5.2. To obtain a soybean 11S globulin protein that is an insoluble fraction by separating the water-soluble fraction, and (3) adjusting the water-soluble fraction to pH 4 to 5.5 at 40 to 65 ° C. After heating, the pH is adjusted to 5.3 to 5.7, and the water-soluble fraction is separated to obtain a non-7S-11S-acid-precipitated soybean protein that is an insoluble fraction. (4) The pH is adjusted to 5.3 to 5.7. Adjusting the separated water-soluble fraction to pH 4-5 to obtain soybean 7S globulin protein, which is an insoluble fraction;
• Processed soybean power according to 30 Adjust the prepared soy milk to pH 5.2-6.4, A method for producing a non-7S-11S-acid-precipitating soybean protein, characterized in that the water-soluble fraction obtained by separating the fraction is adjusted to pH 4 to 5 and the insoluble fraction is recovered;
• a method for fractionating soy protein comprising the following steps: (1) a step of adding water to the processed soybean according to 30 above and separating it into soy milk and okara; (2) the soy milk having a pH of 5.2 to Adjusting to 6.4, and separating the water-soluble fraction to obtain soybean 11S globulin protein, which is an insoluble fraction. (3) Adjusting the water-soluble fraction to pH 4-5 to separate the water-soluble fraction.
• composition for lowering blood cholesterol comprising the non-7S, 11S acid-precipitating soybean protein according to 16.
• composition for lowering blood cholesterol comprising the non-7S′11S acid-precipitating soybean protein according to 20.
• soybean protein can be fractionated with high purity into three fractions of 7S globulin, 11S globulin and lipophilic protein.
• This fractionation method is different from the conventional fractionation method by adding salt, etc., and is a method that mainly adjusts the pH without adding salts, so it is necessary to recover proteins as precipitates. Dilution and desalting operations to achieve a low ion concentration environment are insoluble, and it is an excellent method that simplifies the operation.
• soy milk and a separated soy protein which are excellent in flavor and contain almost no processed soybean power lipophilic protein.
• Soy milk obtained from this processed soybean as a raw material, isolated soy protein, soy 11S protein, soy 7S protein, okara, non-7S'11S acid-precipitating soy protein is disliked compared to conventional soy protein materials It has a refreshing and extremely good flavor, so it is useful as a food material. Power S is even higher.
• the lipophilic protein was confirmed to have a serum cholesterol-reducing effect higher than that of conventional isolated soy protein, a novel material, non-7S, 11S acid-precipitated soybean protein, was used as a health functional ingredient.
• the utility value is high.
• 7S globulin is also called ⁇ -conglycinin, and is generally a glycoprotein composed of three subunits ( ⁇ , ⁇ , j8). good. These subunits are randomly combined to form a trimer.
• the isoelectric point is around pH 4.8 and the molecular weight is about 170,000.
• it may be simply abbreviated as “7S”.
• Soybean 7S protein is a soy protein material with an increased purity of 7S.
• 11S globulin is also called glycinin, and an acidic subunit and a basic subunit are linked by a disulfide bond to form a 12-mer consisting of 6 molecules.
• the molecular weight is about 360,000. Hereinafter, it may be simply abbreviated as “11S”.
• Soybean 11S protein is a soy protein material having a higher purity of 11S.
• Both 7S and 11S are acid-precipitated soy proteins and are the main storage proteins stored in soy protein bodies.
• “acid-precipitating soybean protein” is a protein having a property of being precipitated by adjusting the pH of a solution such as defatted soymilk to the acidic side (pH 4 to 6).
• the protein strength contained in isolated soy protein S corresponds to this, and it does not acidify during the production of isolated soy protein, but does not include protein in whey.
• 7S and 1 IS may be abbreviated as “MSP”.
• Lipophilic Proteins is a group of minor acid-precipitating soy proteins other than 7S and 11S among soy acid-precipitating soy proteins, and polar lipids such as lecithin and glycolipids. Many are accompanied. Hereinafter, it may be simply abbreviated as “LP”.
• This LP mainly contains proteins with estimated molecular weights of 34 kDa, 24 kDa, and 18 kDa as determined by SDS-polyacrylamide electrophoresis, lipoxygenase, ⁇ -conglycinin, and many other miscellaneous proteins (see Figure 2, Lane 3). ).
• LP has the property that it is harder to be stained by SDS electrophoresis than 7S and 11S, so the actual situation has not been clearly recognized. Therefore, it is published as a single band of 7S and 11S in the conventional literature! In fact, a lot of LP is often mixed in the band of SDS electrophoresis.
• LP is a mixture of miscellaneous proteins, it is difficult to identify all the proteins, but it can be fractionated by the dissolution behavior shown in (Method 1) and (Method 2) below. .
• Non-7S'11S-acid-precipitating soy protein refers to a soy protein material with an increased LP purity. Hereinafter, it may be simply abbreviated as “LP-SPI”.
• rpDIJ is an abbreviation for Protein Dispersibility Index, and measures the Dispersfcle protein in soybean products described under the AOCS official method (BalO-65) under certain conditions. Is an index obtained by In contrast to the gentle agitation method that seeks “NSI” (AOCS official method Ball-65), the high agitation operation used in this method generally gives higher numerical results.
• NMI AOCS official method Ball-65
• water was added to soybean, and after stirring with a mixer, the amount of nitrogen in the centrifugal supernatant was measured to determine the ratio to the amount of nitrogen in soybean. The higher the number, the higher the protein solubility of soybean. If it becomes difficult to dissolve soy protein due to heat treatment, the PDI value decreases.
• the "selective water insolubilization index” is an index that numerically indicates how selectively the LP in the processed soybean of the present invention is water-insoluble, and in the water-soluble fraction of soybean.
• the ratio of LP nitrogen (%) and MSP nitrogen (%) to total nitrogen is expressed as “LPZMSP”.
• the first aspect of the present invention is a processed soybean containing a protein and an okara component, having a PDI of 40 or more and less than 80, and wherein a lipophilic protein among the contained proteins is selectively water-insolubilized.
• a processed soybean containing a protein and an okara component, having a PDI of 40 or more and less than 80, and wherein a lipophilic protein among the contained proteins is selectively water-insolubilized.
• the raw soybean used in the present invention contains at least a protein and an okara component, and as long as the oil body that stores lipid exists, the varieties are deficient in LP or the amount thereof is extremely reduced. Since there are no soybean varieties, any variety of soybeans can be applied to the present invention without any particular limitation.
• soybeans rich in 7S there are also soybeans rich in 7S, soybeans rich in 11S, or soybeans in which specific components such as lipoxygenase-deficient varieties have been changed by breeding and genetic recombination techniques, and these can also be used as raw materials.
• 11S-rich soybean that is, 7S globulin-deficient soybean is used as a raw material
• the acid-precipitated soybean protein is mainly composed of 11S and LP.
• the LP of the present invention is selected. Water-insoluble technology can be used.
• LP contains a lot of proteins derived from oil bodies
• soybeans may be used regardless of whether or not the hypocotyl is removed or whether or not the outer skin is removed.
• soybean protein materials such as isolated soybean protein, soybean 7S protein, and soybean 11S protein are prepared using the processed soybean of the present invention as a raw material
• the purity of the protein is affected if lipid is contained. Therefore, it is preferable to use defatted soybeans as raw soybeans.
• a product defatted with an organic solvent such as hexane or a product whose oil content has been reduced by pressing or the like can be used.
• the form of the raw material soybean is not particularly limited, but it is more preferable that the powder be pulverized and have a maximum particle size of 500 m or less, more preferably 300 m or less, and even more preferably 100 m or less. The end is appropriate.
• protein modification in the raw soybean has not progressed extremely before the processing of the present invention. It is preferable that PDI showing a desired protein extraction rate is 60 or more.
• the moisture content of this soybean is preferably 2 to 15%, more preferably 5 to 10%.
• the processed soybean in which the PDI of the present invention is 40 or more and less than 80, and LP is selectively water-insoluble among the contained proteins, in other words, 7S and 11S of acid-precipitable soybean proteins are selectively used. It is characterized by low denaturation.
• LPZMSP selective water insolubility index
• LPZMSP which can be said to have been selectively insoluble in LP, is 45% or less, it is sufficient to obtain a soy protein fraction, more preferably 35% or less, and even more preferably 3%. 0% or less is appropriate.
• the processed soybean itself cannot be identified by direct analysis, but it can be identified by analyzing the water-soluble fraction obtained by extracting water from the processed soybean, that is, LPZMSP contained in soy milk. .
• LPZMSP is obtained after obtaining a water-soluble fraction from potato soybean by the following method 1. It can be calculated by fractionating into LP fraction and MSP fraction by Method 2 and calculating the nitrogen content of each fraction by Kjeldahl method.
• Insoluble fraction C is recovered by centrifugation at 1000G for 10 minutes. Furthermore, to this insoluble fraction C, add 1M Na2S04 (containing 20 mM mercaptoethanol) solution 5 times the sample processed soybean of Method 1 and stir well. Separate into D and insoluble fraction D. Repeat the same procedure for this insoluble fraction D, separating it into water-soluble fraction E and insoluble fraction E. The insoluble fractions D and E are combined as the LP fraction, and the water-soluble fractions D and E are combined as the 7S and 11S fractions (MSP fraction). The operating temperature is 10 ° C to 25 ° C. Measure the nitrogen content of the LP fraction and MSP fraction obtained as described above by the Kelder method, and measure the ratio of both.
• the method for obtaining processed soybeans with the property that LP is selectively water-insolubilized is not particularly limited as long as the selective water-insolubility index (LPZMSP) satisfies the condition of 45% or less.
• LZMSP selective water-insolubility index
• soybean heating may be performed by a dry heating method using a roasting device, a hot air heating device, a microwave heating device, a humidifying heating device, a steaming device, or steam heating.
• a wet heating method using an apparatus or the like can be employed without any particular limitation. However, it is better to avoid heating when water is soaked in soybeans, as protein will be extracted.
• soybean is enclosed in a sealed tank and the inside of the jacket covering the outside of the sealed tank is heated so that the product temperature is about 70 to 95 ° C in an atmosphere with a relative humidity of 90% or more can be adopted.
• the heating temperature and time conditions are not particularly limited as long as LP insolubles are selective, but the temperature is usually set to 60 to 95 ° C, and the time is 1 minute to 10 hours. It is appropriate to perform between.
• alcohol modification When alcohol modification is used as another embodiment of the modification method, it is equal to or less than 1 part by weight, preferably 2 to 100 parts by weight, more preferably 8 to 20 parts by weight with respect to the raw soybean containing the protein and the sugar component. More preferably, a method of adding and impregnating 10 to 15 parts by weight of a polar alcohol solution is preferred. According to this method, LPZMSP can be easily reduced to 30% or less, and only LP can be selectively insoluble in water more efficiently.
• a polar solvent suitable for promoting selective water insolubilization of LP a polar alcohol solution (methanol, ethanol, propanol, isopropanol, etc.) can be used.
• a polar alcohol solution methanol, ethanol, propanol, isopropanol, etc.
• Water may be pure water, or an acid aqueous solution (hydrochloric acid aqueous solution, carbonic acid aqueous solution, citrate aqueous solution, etc.), alkaline aqueous solution (hydroxy sodium hydroxide solution, sodium bicarbonate, etc.), etc. may be used.
• the concentration of the polar solvent solution is 5 to: LOO% is preferred, and 50 to 80% is more preferred. If the concentration of the polar solvent solution is too low or too high, water insolubility caused by LP modification will be insufficient.
• the method for adding the polar solvent solution can be carried out by, for example, a method of spraying the powder by spraying or a method of dropping, but is not particularly limited.
• a stirrer such as a single or a high-speed stirrer can be used.
• the heating temperature is preferably 30 to 95 ° C, more preferably 40 to 90 ° C, as the product temperature of soybean.
• the calorie temperature time is preferably 5 to 100 minutes, more preferably 10 to 60 minutes.
• the order of these steps is not particularly limited. However, after adding and mixing the polar solvent, the heating treatment is performed or the polar solvent is added. It is preferable to perform heat treatment while mixing.
• the amount of the polar solvent added can be reduced, the removal process of the polar solvent after the treatment is extremely easy compared with the conventional alcohol cleaning method, etc., which is advantageous for establishing an efficient manufacturing process. It is.
• Most of the polar solvent remaining in the processed soybean can be volatilized by heating treatment, and this can be directly subjected to the extraction process. If it is treated at a temperature of 40-60 ° C and reduced pressure (about 1 lOmmHg) for 10-60 minutes, it can be completely volatilized and returned to the weight of soybean before addition.
• volatilized polar solvent can be reused if recovered by distillation, This is advantageous in the manufacturing process.
• the following invention is used as a raw material for fractionation of one or more acid-precipitating soy proteins selected from 7S, 11S and LP, which are selectively water-insoluble in LP. This is a common technical feature.
• a fraction enriched with at least one acid-precipitating soy protein selected from the group consisting of 7S globulin, 11S globulin, or lipophilic protein strength using the soymilk or okara prepared with the above-described processed soybean strength as a raw material. It is an invention for collecting and obtaining fractionated soybean protein.
• the soy milk of the present invention is a soy milk made from the processed soybean.
• it is soy milk made from baked soybeans obtained by the above alcohol modification treatment.
• the soy milk of the present invention is not particularly limited as long as it uses the above-described processed soybean as a raw material, but it is extracted with an aqueous solvent such as water or an aqueous alkaline solution, and separated into soy milk and okara by centrifugation. It is obtained by collecting the soluble fraction.
• an aqueous solvent such as water or an aqueous alkaline solution
• the addition amount of the aqueous solvent is preferably 6 to 12 times by weight, more preferably 7 to 9 times by weight with respect to the processed soybean. If the amount of the aqueous solvent added is too small, the viscosity will increase, and if it is too large, the solution will become a dilute solution and the recovery efficiency will deteriorate.
• the temperature during extraction is preferably about 4 to 50 ° C, more preferably about 10 to 30 ° C. If the temperature is too high, the LP will be easily dissolved. Conversely, if the temperature is too low, the extraction efficiency will deteriorate. By removing the okara, which is insoluble at a pH near pH 9 to 9, by centrifugation, etc. To do. The operation of adding 4 to 6 times the amount of water to the resulting okara and further extracting it to increase the amount of recovered soy milk may be repeated.
• soy milk made from the processed soybean of the present invention may be commercialized as it is, or may be further processed into concentrated soy milk or powdered soy milk, or prepared by adding an appropriate raw material. It can also be processed into soy milk.
• the protein composition of the obtained soymilk has a very characteristic composition unlike ordinary soymilk, and LPZMSP with a low LP content is 45% or less, preferably 35% or less, more preferably It is preferably 30% or less, more preferably 28% or less, and most preferably 23% or less.
• the soy milk can be used for the production of the following fractionated soy protein.
• the soy milk extracted from soy milk by a normal manufacturing method or soy that has been subjected to non-selective protein insolubilization treatment has an LPZMSP ratio of more than 45% (see Table 1).
• the fractionation ability to soybean 7S globulin protein and soybean 11S globulin protein can be achieved by simple means only by adjusting H.
• the isolated soy protein of the present invention is characterized by using the soy milk obtained from the above-described processed soybean strength as a raw material, and other than that, it can also be produced by a known production method that is usually performed.
• an acid hydrochloric acid, sulfuric acid, etc.
• an acid is added to the soymilk of the present invention to acidify the pH of the soymilk.
• the pH should be adjusted to around the isoelectric point of soy protein. It is preferable to adjust the pH to 4.2 to 5.2.
• acid-precipitating soy protein becomes insoluble in this pH range and becomes a precipitate. This is recovered by centrifugation and neutralized by adding alkaline water such as caustic soda to prepare a neutralized solution of soybean protein to obtain separated soybean protein.
• alkaline water such as caustic soda
• This can be sterilized and dried if desired and used as it is in the form of a powder, or it can be used as a separated soy protein for various conventional food applications by adding a suitable raw material for preparation. .
• the defatted soybean of the present invention can be acid-washed and then extracted with water to obtain isolated soybean protein.
• the obtained isolated soy protein is characterized by having a crude protein content of 90% by weight or more and a flavor superior to that of conventional isolated soy protein due to its low LP content.
• the isolated soy protein provided as a soy protein material is generally heat sterilized in the final production process, so 7S and 11S are heat-denatured together with LP. Therefore, it is difficult to measure the LP content by separating LP into 7S and 11S by the methods 1 and 2 described above.
• SDS-polyacrylamide gel electrophoresis which is a general method for measuring protein composition, has the property that LP is difficult to be stained with CBB, which is also an accurate measurement. It is difficult to determine.
• This method can be applied not only to isolated soybean protein but also to various fractionated soybean proteins such as soybean 7S protein, soybean 11S protein, and LP-SP I.
• LCI Lipophilic Proteins Content Index
• P34 LP major component, 34kDa protein
• Lx LP main component, lipoxygenase
• the isolated soybean protein obtained by the present invention has an LCI of 38% or less, preferably 35% or less, more preferably 30% or less, and further preferably 25% or less. If the LCI exceeds 38%, it will approach the LCI value of the soy protein isolated by the conventional method, and the quality will not change. On the other hand, the smaller the LCI strength, S, the better the flavor.
• soybean protein of the present invention As described in the prior art, various methods have been used to fractionate acid-precipitable soybean protein into 11S and 7S. Soybean 11S protein of the present invention and this soybean 7S protein are both of the present invention.
• the processed soybean power is obtained by fractionating the prepared soy milk.
• the soybean 11S protein of the present invention is prepared by adjusting the soy milk of the present invention to a specific pH and recovering the insoluble fraction produced, and if necessary, neutralizing, sterilizing, drying, and using as it is in the form of a powder.
• a fractionated soy protein material can be obtained with high purity and efficiency by adding a suitable preparation material to prepare a preparation.
• the electrophoresis pattern of the obtained fraction has a low LP content (Fig. 4) and an excellent flavor, similar to the electrophoresis pattern of the soymilk of the present invention.
• a reducing agent such as sodium sulfite may be added to soy milk and then adjusted to the above pH range. This has the advantage that the separability is further improved. In the case of this soymilk, good separation is possible at about ImM.
• Non-Patent Documents 2 and 6 the property of 11 S that cools and settles, which is a conventional 11S separation method, may be used. That is, by adding a reducing agent to the soymilk of the present invention, adjusting the pH of the soymilk to 6.1-6.5, cooling to 4-6 ° C and leaving it for half a day, and collecting the produced precipitate, Is recovered.
• the reducing agent is typically 10mM. In the case of this soymilk, good separation is possible at about ImM.
• the obtained soybean 11S protein has a high purity of 11S of 75% by weight or more, further 85% by weight or more, and further 90% by weight or more, it can be utilized by utilizing the characteristics unique to 11S.
• 11S has low viscosity and high gel strength due to heating, so it can be used, for example, as a gelling agent, and as a substitute for egg white, and tofu can be hardened. It can be used.
• the LCI value is 30% or less, more preferably 25% or less, and further preferably 20% or less, and the flavor is excellent with a very low LP content.
• the soybean 7S protein of the present invention is an insoluble fraction produced by adjusting the pH of the water-soluble fraction solution after fractionation of the above 11S to pH 4 to 5.5, preferably 4.3 to 4.8 with an acid. It can be obtained by recovering the fraction, sterilizing and drying without force to neutralize it as desired, and using it in the form of powder as it is or by adding an appropriate drug substance . In this case, the purity of 7S in soybean 7S protein is at least 38% or more, further 40% or more, further 50% or more, and further 60% or more. [0085] To further increase the degree of purification of 7S, LP can be removed in advance as an insoluble fraction prior to the above step.
• the pH of the water-soluble fraction obtained at the time of preparation of the soybean 11S protein of the present invention is adjusted to pH 4 to 5.5, preferably pH 4.8 to 5.2 [after adjustment and heating at 40 to 65 ° C.]
• proteins other than 7S LP-based
• proteins other than 7S become insoluble and can be removed as an insoluble fraction. If it is not necessary to simultaneously collect 7S and 11S from soymilk, only 7S is fractionated directly using the soymilk of the present invention instead of the above water-soluble fraction, and 11S and LP as insoluble fractions. It can also be excluded.
• the pH of the water-soluble fraction after removal of the insoluble fraction is adjusted to pH 4-5, preferably 4.3-4.8 with acid, and the insoluble fraction produced is recovered, so that the soybean with higher purity can be obtained. It is reasonable to obtain 7S protein.
• the method for preparing soybean 7S protein is not limited to this method, and a conventional 7S fractionation method may be used if desired.
• a conventional 7S fractionation method may be used if desired.
• remove 11S globulins! Add soymilk with NaCl to a concentration of 0.25M, adjust the pH to 5.0 to remove insoluble fractions, There is a method to recover the precipitate that is formed by adding 3 volumes of water to bring the pH to 4.5.
• Non-Patent Document 5 sulfuric acid is added to soy milk excluding 11S globulin, the pH is adjusted to 2.8 to 3.5, and the resulting precipitate is removed, and then twice the volume of water is added thereto. In addition, there is a method of recovering the precipitate formed at pH 4.5.
• the 7S purity of the high-purity soybean 7S protein obtained as described above is at least 80% or higher, so that it is possible to utilize the characteristics unique to 7S.
• it can be used for nutritional functional agents such as blood neutral fat reducing agents and body fat reducing agents, and highly viscous materials.
• the LCI is 30% or less, more preferably 25% or less, still more preferably 20% or less, and the flavor is excellent with a very low LP content.
• the okara of the present invention is an okara obtained by using the processed soybean of the present invention as a raw material, extracting this with water, and collecting the insoluble fraction. Since this potato soy is an LP that is selectively water-insoluble, extraction with water as described above will extract 7S and 11S to the soy milk side, and LP will be mainly distributed to the okara side. Drawn.
• this Okara is characterized by its richness in LP.
• LP content is about 10 to 20% by weight in the dry solid content of ordinary Okara, but this Okara is 35 to 60% by weight.
• LP is particularly excellent in blood cholesterol lowering effect among soy acid-precipitating soybean proteins. Therefore, it is possible to give high added value to the okara that is generally disposed of as a by-product of soymilk.
• This okara is obtained by collecting the insoluble fraction separated during the production of the above-mentioned soymilk by centrifugation or the like. If desired, it can be made into various forms of products by processing such as sterilization, freezing, crushing, and drying.
• LP is a force that was thought to be a component that contributes to the flavor deterioration of conventional soy protein materials. By separating this into high purity, non-7S'11S-acid-precipitated soy protein It can be used for applications that make use of the unique characteristics of LP.
• the LP-SPI of the present invention can be obtained by fractionation by two methods: using okara prepared from the above-mentioned processed soybean as a raw material and using soymilk as a raw material.
• the first LP is obtained by fractionating the above-prepared okara that has been prepared from the above-mentioned potato soybean power, and the oil content extracted with a solvent having a 2: 1 volume ratio of black mouth form and methanol is 7% or more, The content is preferably 8% or more. Preparation examples are shown below.
• LP is selectively contained in the processed soybean of the present invention in an insoluble state, it can be fractionated from the above-mentioned okara, which is a residue obtained by extracting the soy milk.
• the fraction can be obtained by adding water to okara and extracting by heating and collecting the extract.
• the amount of water is preferably 50 to 500 parts by weight of water per 100 parts by weight of Okara.
• the heating temperature is preferably 100 to 150 ° C.
• the heating time is preferably between a few seconds and a few minutes.
• the extract obtained by the above method can be provided as LP-SPI having an LCI of 50% by weight or more, preferably 60% by weight or more. If desired, add acid to the extract and adjust the pH to 4-5. Preferably, it is adjusted to 4.3 to 4.8, and the precipitate formed can be recovered to obtain LP-SPI with higher purity. This is neutralized with a soda solution to prepare a neutralized solution, sterilized and heated.
• the LP-SPI obtained by the above method can be provided as a high-purity product having at least LCI of 60% by weight or more, preferably 65% by weight or more.
• the second LP is obtained by fractionating the soymilk prepared from the above-mentioned soybean soy strength, and the oil content extracted with a solvent having a volume ratio of black mouth form and methanol of 2: 1 is 7% or more.
• the fractionation method is characterized by containing 8% or more. Preparation examples are shown below.
• processed soybean power Adjusted soy milk to pH 5.2 to 6.4 and separated the insoluble fraction to obtain a water-soluble fraction.
• the pH is adjusted to 4 to 5.5, heated at 40 to 65 ° C, and then the insoluble fraction produced when the pH is adjusted to 5.3 to 5.7 is recovered, whereby LP can be fractionated with high purity.
• the fraction obtained by the above method is neutralized with a soda as needed to prepare a neutralized solution, sterilized and heated and dried.
• the LP-SPI obtained by the above method can be provided as a high purity product having at least LCI of 60% by weight or more.
• the oil content (hereinafter referred to as “chroma oil content”) extracted with a 2: 1 solvent of chloroform-form: methanol in the protein is 7% by weight or more, preferably 8-15% by weight, more preferably 9%. This can be done with a force of ⁇ 15% by weight.
• the LP-SPI ether extract oil content is 2% or more, the above-mentioned numerical force must also be deducted from the ether extract oil content.
• the polar lipids to be released are mainly composed of lecithin and glycolipid.
• the chromed oil content of conventional unisolated soybean protein is about 4-5% by weight, and high-purity soybean 7S protein and soybean 11S protein are only 3% or less.
• LP-SPI has blood cholesterol lowering activity even if it is fractionated by any of the above methods.
• LP-SPI is isolated soy protein, soy 7S protein, soy 11S protein. It has been confirmed that it has a remarkably strong blood cholesterol-reducing action compared to the above. It has also been confirmed that 7S having a high purity, that is, low LP, as used in the present study hardly shows a blood cholesterol lowering effect.
• LP-SPI exhibits a stronger cholesterol-lowering action by coexistence of LP and clometa extract and their presence as a complex.
• the content of LP-SPI added to the composition for lowering blood cholesterol of the present invention varies depending on the amount of the composition, and can be appropriately set. In general, a person skilled in the art may set the content in the composition in consideration of the intake amount of the composition per day so that the intake amount of the active ingredient per day can be taken. For example, if the daily intake of LP-SPI is set to 4.5 g, and the daily intake of the composition is 10 g, the active ingredient content in the composition is 45% by weight. What should I do?
• the daily intake of LP-SPI of the present invention is not particularly limited, but can be 4 to 10 g.
• the composition for lowering blood cholesterol of the present invention can be used in combination with a material that is said to have a blood cholesterol lowering action.
• a material that is said to have a blood cholesterol lowering action for example, isoflavones, soy milk, separated soy protein, concentrated soy protein, lecithin, lactic acid bacteria, polyphenols, polysaccharides and the like can be used in combination.
• the form of the composition for lowering blood cholesterol of the present invention can be an agent or a food.
• an agent in the case of an agent, it can be prepared in various dosage forms. That is, in the case of oral administration, it can be administered in the form of solid preparations such as tablets, hard capsules, soft capsules, granules or pills, or liquid preparations such as solutions, emulsions or suspensions. In the case of parenteral administration, it is administered in the form of an injection solution or a suppository.
• additives that are acceptable for formulation, such as excipients, stabilizers, preservatives, wetting agents, emulsifiers, lubricants, sweeteners, coloring agents, flavoring agents, tonicity adjusting agents.
• a buffer, an antioxidant, a pH adjuster, etc. can be used in combination.
• soft drinks, dairy products, soy milk, fermented soy milk, soy protein drinks, tofu, natto, fried chicken, deep fried, gandou, hamburger, meatballs, fried chicken, nuggets It can be blended into various foods such as various prepared foods, baked goods, nutrition bars, cereals, rice cakes, gums, jelly and other sweets, tablets, breads, cooked rice.
• LP-SPI is included as an active ingredient in advertising media such as food packaging and pamphlets, and that this has the effect of reducing blood cholesterol. It can also be used as health foods such as Japanese specific health foods.
• the advantages of the present invention are as follows. First, by selectively subjecting LP to water insolubilization, the fractionation of 7S, 11S, and LP, which conventionally required complicated operations, is increased. Purity, efficiency and simplicity. By using the processed soybean of the present invention as a raw material, it becomes possible to fractionate the mixture of 7 S and 11 S with high purity by simply precipitating at a specific isoelectric point. Moreover, it becomes possible to fractionate LP which has not been recognized so far with high purity. LP-SPI containing LP in a high degree has a blood cholesterol lowering action stronger than isolated soy protein, and can be provided as a novel soy protein material.
• the second advantage of the present invention is that the flavor of existing soy protein materials such as soy milk and isolated soy protein can be improved. That is, the processing of soybean of the present invention can selectively insolubilize LP and lead to a state in which lipids associated with LP are difficult to be extracted. As a result, the flavor of the extracted soy milk and the various soy protein ingredients prepared from the soy milk is rated. Improved to the stage.
• the processed soybean of the present invention is one in which the number of bacteria in the soybean is reduced by the processing treatment. This suppresses the growth of bacteria in the water-based cache process, and has an advantage not only in flavor but also in satellite.
• the obtained soymilk has a good flavor, it can be provided as a high-quality soymilk material. Furthermore, the separated soy protein, soy 7S protein, and soy 11S protein produced by fractionation are all very good in color and flavor even when subjected to heat treatment such as retort sterilization. Will not darken or taste bad.
• the deactivation and sterilization of the enzymes involved in the oxidative degradation of the present invention using polar alcohol can be applied to whole-fat soybeans as well as defatted soybeans. It is also very effective in improving the flavor of the soy milk prepared.
• Hydrated ethanol (10%, 50%, 60%, 70%, and 80%) was mixed with 1 kg of low-denatured defatted soybean (PDI: 83, moisture 7.0%) filled in a sealed container while spraying 100 g each.
• the outside of the sealed container was heated so that the temperature of the defatted soybean reached 70 ° C and maintained for 30 minutes.
• the defatted soybean was taken out from the container and allowed to cool to prepare a processed defatted soybean.
• the PDIs were 71, 67, 64, 65, and 64, respectively.
• Calorie defatted soybean was prepared in the same manner as in Example 1 except that the spray amount of hydrous ethanol (70%) was increased to 150 g. This PDI was 72.
• Calorie defatted soybean was prepared in the same manner as in Example 1 except that the spray amount of hydrous ethanol (70%) was increased to 200 g. This PDI was 45.
• the outside of the sealed container was heated to maintain the temperature of the defatted soybean (PDI: 83, moisture 7.0%) 1 kg in an atmosphere with a relative humidity of 90% or more so that the product temperature of the defatted soybean was 75 ° C and maintained for 30 minutes.
• Degreased soybeans were taken out from the container, and processed defatted soybeans were prepared. This PDI was 73.
• a processed defatted soybean was prepared in the same manner as in Comparative Example 1 except that the temperature of the defatted soybean was heated to 85 ° C and maintained for 60 minutes. This PDI was 66.
• Caloe defatted soybean was prepared in the same manner as in Example 1 except that the spray amount of hydrous ethanol (70%) was reduced to 30 g. This PDI was 79.
• Processed defatted soybeans were prepared in the same manner as in Example 1 except that the spray amount of hydrous ethanol (80%) was increased to 1.5 kg and the heating maintenance time was extended to 60 minutes. This PDI was 32.
• the nitrogen content of the obtained whey fraction, okara fraction, LP fraction and MSP fraction was analyzed by the Kjeldahl method, respectively, and the total nitrogen content in defatted soybean was 100%.
• the nitrogen transfer rate (%) to each fraction was calculated.
• the nitrogen ratio of LP fraction and MSP fraction (LPZ The selective water insolubility index which is MSP) was calculated.
• soy milk was also extracted and analyzed for low-denatured defatted soybeans that were not processed. The results are shown in Table 2.
• Example 3 the rate of nitrogen transfer to MSP decreased as in Example 3, and some of the forces decreased in yield.
• the nitrogen transfer rate was close to 48%. In other words, it was confirmed that 7S and 11S were extracted to the soy milk side with a high yield.
• Example 6 and Comparative Examples 1 and 2 From the results of Examples 1 to 3, Example 6, and Comparative Examples 1 and 2, the higher the ethanol concentration and the greater the added amount, the lower the PDI, and the higher the degree of modification of defatted soybeans.
• the amount of ethanol added increased to some extent, the tendency of the nitrogen transfer rate to the LP fraction decreased, and conversely the nitrogen transfer rate of MSP tended to decrease.
• LPZMSP increased and Comparative Example 2 showed the same value as the control.
• PDI had a value similar to that of ethanol addition, and LP was selectively insoluble in water.
• Example 6 which was heated strongly, the rate of nitrogen transfer to the MSP fraction was decreasing, and the LPZMSP ratio was slightly higher than in the case of ethanol treatment. In comparison between wet heat and ethanol addition, the ethanol addition force LPZMSP becomes smaller and LP tends to be more selectively water-insoluble.
• the processing conditions for defatted soybeans that selectively insolubilize LP with water were such that PDI was 40 or more and less than 80 and LPZMSP was 45% or less.
• control by wet heating and a method of adding 5 to 100% LOO wt% of water-containing ethanol having a concentration of 5 to 100% to soybeans were suitable.
• LPZMSP could be reduced to 35% or less.
• Each soymilk was adjusted to pH 4.5 with hydrochloric acid, and the precipitate was collected by centrifugation to remove the whey fraction. Further, the collected precipitate was neutralized with sodium hydroxide and watered to a concentration of 3%. The neutralized solution was boiled for 10 minutes and then cooled to room temperature to prepare a separated soy protein solution for flavor test.
• the separated soy protein solution is analyzed by SDS polyacrylamide gel electrophoresis under the conditions shown in Table 1 according to the LP content estimation method described above, and the LCI value (see Equation 1), which is the estimated LP content, is obtained. It was.
• the flavor of each soymilk and separated soy protein solution obtained was tested by 10 panelists. The score was 10 out of 10 points, and the higher the score, the less bad flavor. As a standard, 5 untreated preparations were scored. The average score is calculated by dividing the total score by the number of panelists.
• Example 2 70% ethanol 8.9 9.4 23 Clean Example 3 70% ethanol 8.5 9.3 26
• Example 5 Moist heat 6.0 6.8 36 Roast odor Example 6 70% ethanol 5.9 6.0 34
• soybean 11S protein The soymilk prepared from each processed defatted soybean in Comparative Experimental Example 1 was adjusted to pH 5.8 with hydrochloric acid, and the resulting precipitate was collected by centrifugation at 1000 G for 10 minutes. This insoluble fraction was designated as soybean 11S protein.
• the purity of soybean 11S protein was 75% or higher, and in Examples 1 to 3, the purity was 90% or higher, and the recovery rate was equivalent or higher.
• the purity of soybean 7S protein was 38% or more in all Examples, 50% or more in Examples 1 to 3, and some Examples exceeded 60%.
• the flavor of these proteins was very good, and the taste was very good!
• Comparative Example 1 the purity of soybean 11S protein was similar to that of the Example, but the recovery rate was 6%, which was considerably low. This is thought to be because the selective water insolubility of LP with a large amount of 7S and 11S transferred to Okara was insufficient.
• the flavor of this protein was excellent, and it was evaluated as ⁇ with less disgusting taste.
• the oil contained in the solid content of this protein is 1% of the oil extracted with ether, 11% of the oil extracted with a mixed solvent with a 2: 1 ratio of formaldehyde to methanol, and polar It was shown that many LPs showing affinity for lipid were contained.
• the resulting LP-SPI contained a high amount of LP, which was previously thought to be a causative component of the off-flavor of soy protein isolate, and was expected to have a bad taste. It had a good flavor with no bad flavor.
• the LCI value at this time was 72%.
• Insoluble fraction A obtained by the same method as in Example 7 was collected, and this fraction was designated LP-SPI.
• the oil contained in the solid content of this protein is 1% of the oil extracted with ether, and 9% of the oil is extracted with a mixed solvent with a 2: 1 ratio of formaldehyde to methanol. It was shown that many LPs having affinity for polar lipids were contained.
• the obtained LP-SPI like the LP-SPI of Example 7, had a surprisingly good taste with no bad taste.
• the LCI value at this time was 71%.
• vitamin-free casein made by Oriental Yeast Co., Ltd., hereinafter referred to as “casein”.
• the rats were fasted for 6 hours from 8:00 am and then opened under Nembutal anesthesia, and blood was collected from the abdominal aorta. The blood was centrifuged at 3000 RPM for 15 minutes after heparin treatment, and the resulting plasma was used as a blood sample to measure blood total cholesterol (TC) and fecal steroid excretion.
• TC total cholesterol
• TC was measured using Fuji Dry Chem 5500 (Fuji Film Co., Ltd.).
• fecal steroid excretion fecal samples were collected, freeze-dried and crushed for 3 days immediately before slaughter.
• Excreted neutral and acidic steroids by Mathtinen et al. (Miettinen, TA; Ahrens, EH Jr .; Grundy, SM Quantitative isolation and gas—liquid chromatogra phic analysis of total dietary and fecal neutral steroids. J. Lipid Res., 6, 411-424, 19 65.), Grundy et al.
• Table 6 shows the results of changes in cholesterol levels and fecal total steroid excretion in rats fed the 2-week test diet.
• LP-SPI intake was found to have a significantly lower blood cholesterol lowering effect than isolated soy protein and soy 7S protein, and also tended to decrease with respect to soy 11S protein. That is, it was confirmed that LP-SPI fractionated by the method of the present invention has a stronger blood cholesterol lowering effect than the conventional soybean protein material.
• LP-SPI was washed with ethanol, and the effect of removing the clometa extract on blood cholesterol reduction was examined.
• LP-SPI produced in the same manner as in Example 8 was washed once with 10 volumes of 70% ethanol, then once with 3 volumes of 70% ethanol, and then with 2 volumes of 99.5% ethanol. Washed with washing. After drying overnight at room temperature, drying at 60 ° C for 1 hour, ethanol washing LP-S Obtained PI (LP-EW). The content of Crometa extract in LP-EW was 1.4%.
• lipid was dissolved in soybean oil shown in Table 5, and a sample (LP-EW + Lipid) mixed with LP-EW in the test meal again was obtained.
• a test diet (Table 5) was prepared by substituting 10 wt% of the protein source with LP-SPI, LP-EW, LP-EW + Lipid, respectively, using a diet containing 20 wt% casein as a control. Animals were ingested 2g daily as protein in the following manner. Each protein intake group was a casein group (control group), LP-SPI group, LP-EW group, LP-EW + Lipid group, and model animals were 6-week-old WISTAR male rats (sold by Japan SLC Co., Ltd.). 24) were used. After one week of preliminary breeding, each group was divided into 6 animals so that the average body weight between groups was almost the same, and the test food was raised for 2 weeks. Table 7 shows the results of changes in blood cholesterol levels in rats fed with test samples for 2 weeks after the end of the test period.
• LP-SPI showed a tendency to lose its powerful blood cholesterol-reducing effect even if it was mixed again when LP-SPI was removed with ethanol. It was. Therefore, the LP-SPI of the present invention has a stronger blood flow due to coexistence and complexation of a chromate extract such as lecithin, which has a higher affinity for LP, than the presence of LP alone. It is thought to show an effect of reducing medium cholesterol.
• soybean protein By using the processed soybean of the present invention as a raw material, it is possible to fractionate soybean protein into 7S globulin, 11S globulin, and lipophilic protein, respectively, with high purity and easily, and the conventional fractionation method This can greatly improve the complicated manufacturing process.
• soybean 7S protein, soybean 11S protein, and non-7S'11S-acid-precipitating soybean protein it becomes possible to produce foods with more vigorous physical properties and nutritional physiological functions.
• 7S-11S-acid-precipitated soy protein has been commercialized so far, and it is a novel soy protein material that has a higher cholesterol lowering effect than conventional soy protein isolates. Use for nutrition improvement is expected.
• soy milk, isolated soybean protein, soybean 7S protein, soybean 11S protein, okara, non-7S ⁇ 11S-acid-precipitated soy protein obtained in the present invention has a very flavor compared to conventional soybean protein materials. Since it is good, it is highly useful in improving the quality of conventional foods that use these.
• Fig. 1 is a graph showing the dissolution behavior of 7S globulin and 11S globulin at each pH.
• FIG. 2 is a drawing-substituting photograph showing migration patterns of SDS-polyacrylamide gel electrophoresis of 7S globulin fraction, 11S globulin fraction, and lipophilic protein fraction.
• FIG. 3 is a drawing-substituting photograph showing the migration pattern of the prepared soybean 11S globulin protein and okara in SDS-polyacrylamide gel electrophoresis in Example 2 and Comparative Example 1;
• FIG. 4 SDS-polyacrylamide gel electrophoresis of each fraction (okara, defatted soymilk, 11S globulin, 7S impurities, 7S globulin, whey, lipophilic protein) prepared with processed defatted soybean power of Example 2 It is the drawing substitute photograph which showed the electrophoresis pattern.

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