WO2010092778A1 - Acid-soluble soybean protein material, and process for producing same - Google Patents

Acid-soluble soybean protein material, and process for producing same Download PDF

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Publication number
WO2010092778A1
WO2010092778A1 PCT/JP2010/000733 JP2010000733W WO2010092778A1 WO 2010092778 A1 WO2010092778 A1 WO 2010092778A1 JP 2010000733 W JP2010000733 W JP 2010000733W WO 2010092778 A1 WO2010092778 A1 WO 2010092778A1
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acid
soybean protein
acidic
protein material
soluble soybean
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PCT/JP2010/000733
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French (fr)
Japanese (ja)
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上山知樹
加藤裕之
坂田哲夫
本山貴康
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不二製油株式会社
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Priority to JP2010550444A priority Critical patent/JPWO2010092778A1/en
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23G—COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
    • A23G3/00—Sweetmeats; Confectionery; Marzipan; Coated or filled products
    • A23G3/34—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof
    • A23G3/36—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof characterised by the composition containing organic or inorganic compounds
    • A23G3/44—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof characterised by the composition containing organic or inorganic compounds containing peptides or proteins
    • 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
    • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00—Working-up of proteins for foodstuffs
    • A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
    • A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
    • A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
    • A23J3/346—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of vegetable proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
    • A23L11/30—Removing undesirable substances, e.g. bitter substances
    • A23L11/31—Removing undesirable substances, e.g. bitter substances by heating without chemical treatment, e.g. steam treatment, cooking
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
    • A23L11/30—Removing undesirable substances, e.g. bitter substances
    • A23L11/33—Removing undesirable substances, e.g. bitter substances using enzymes; Enzymatic transformation of pulses or legumes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • 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
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/52—Adding ingredients
    • A23L2/66—Proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L21/00—Marmalades, jams, jellies or the like; Products from apiculture; Preparation or treatment thereof
    • A23L21/10—Marmalades; Jams; Jellies; Other similar fruit or vegetable compositions; Simulated fruit products
    • A23L21/11—Marmalades; Jams; Jellies; Other similar fruit or vegetable compositions; Simulated fruit products obtained by enzymatic digestion of fruit or vegetable compositions
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L21/00—Marmalades, jams, jellies or the like; Products from apiculture; Preparation or treatment thereof
    • A23L21/10—Marmalades; Jams; Jellies; Other similar fruit or vegetable compositions; Simulated fruit products
    • A23L21/12—Marmalades; Jams; Jellies; Other similar fruit or vegetable compositions; Simulated fruit products derived from fruit or vegetable solids

Definitions

  • the present invention relates to an acid-soluble soybean protein material useful for, for example, acidic foods and drinks.
  • Acidic foods and drinks are characterized by their refreshing flavor, and are typically acidic beverages such as fruit juices, jelly, and yogurts.
  • whey protein which is a main component of milk whey and has high solubility under acidic conditions
  • Separated soy protein which is a soy protein material obtained by separating from soy, has a very low solubility in the pH range around pH 4.5, which is its isoelectric point from around pH 3. Therefore, various attempts have been made to increase the solubility of soy protein containing soy protein in acidic water.
  • the soy protein is heated to 250 to 320 ° F.
  • Patent Document 2 greatly improves the solubility of the soy protein material in the acidic region by a treatment such as phytase treatment of the separated soybean protein followed by high-temperature heat treatment under acidity.
  • Acidic aqueous solutions of these protein materials that dissolve at pH 3.5 have the problem of feeling astringency in the mouth when ingested (Non-patent Document 1).
  • This astringency is induced by isoelectric precipitation of the protein in the mouth, and can be avoided by reducing the molecular weight of the protein.
  • protein hydrolysis under weak acidity reduces astringency, but has a problem of bitterness. Both astringency and bitterness could not be reduced to a level used for food and drink.
  • Patent Document 3 a method of performing acid solubilization treatment by adjusting to weak acidity after performing hydrolysis under neutrality was also studied.
  • a bitterness was hardly generated and astringency was considerably reduced, but further improvement is desired.
  • An acidic soluble soybean protein material that has reduced astringency and bitterness and has reached a level that can be used for acidic foods such as beverages or jelly-like foods is desired.
  • An object of the present invention is to provide an acidic-soluble soybean protein material that is reduced in astringency and bitterness and excellent in flavor for use in acidic foods and drinks.
  • the present inventors have not so far carried out protease degradation of a soy protein-containing solution to a certain degree of degradation when preparing an acidic soluble soybean protein material. It was found that by starting the reaction at pH 3.4 or less, astringency and bitterness are greatly reduced, and this discovered that the content of fractions of 150 kDa or more in gel filtration described later was significantly low, This invention was completed. That is, the present invention (1) In gel filtration using 1 wt% SDS-containing buffer solution (pH 7.0), the content of the fraction with mass number of 150 kDa or more detected at 220 nm is 18% or less and TCA (0.22M) is solubilized.
  • Acid soluble soy protein material with a rate of 45% or less.
  • a method for producing an acidic soluble soybean protein material characterized in that protease treatment is started at pH 3.4 or less, and the TCA (0.22M) solubilization rate is 10% or more and 45% or less.
  • an acidic-soluble soybean protein material suitable for acidic foods and drinks having little astringency and bitterness and excellent flavor.
  • Comparison by gel filtration The product (C) of the present invention is a gel filtration chromatogram showing that the fraction of 150 kDa or more is very small compared to the conventional product (F).
  • Comparison by electrophoresis FIG. 7 is an SDS polyacrylamide gel electrophoresis pattern showing that the 7S globulin and 11S globulin bands do not remain in the product (C) of the present invention as well as the conventional product (F).
  • an acidic soluble soybean protein material is a protein-rich fraction extracted from soybean raw material and separated from okara, and is defined as a dilute acid NSI modified method of 90% or more.
  • one form of the present invention is an acidic soluble soybean protein material, which exhibits a molecular weight distribution described later, has a TCA (0.22M) solubilization rate of 45% or less, and 40% or less. Further preferred. When the TCA (0.22M) solubilization rate exceeds 45%, a lot of bitterness may occur. Further, when the dilute acid NSI is less than 90%, roughness and the like occur, and it is difficult to use as a protein material for acidic foods and drinks.
  • the content of a fraction having a mass number of 150 kDa or more detected at 220 nm in gel filtration using a 1 wt% SDS-containing buffer (pH 7.0) is 18% or less, preferably 14%.
  • the following acidic soluble soybean protein materials An acidic soluble soybean protein material having a mass fraction of more than 150 kDa with a content of more than 18% has a large unpleasant astringency and cannot be used for acidic foods. On the other hand, if it is 18% or less, the astringency is reduced to an unpleasant level or less, and if it is 14% or less, it is almost impossible to feel astringency. Only protein materials that meet the above conditions are the present invention and are suitable for acidic foods and drinks.
  • This gel filtration can be performed as follows. That is, a column of “TSK gel G3000SWXL” ( ⁇ 7.8mm ⁇ 300mm) (manufactured by Tosoh Corporation) was used, and a 50 mM phosphate buffer (pH 7.0) containing 1% SDS and 1.17% NaCl was used as the eluent. The flow rate is 0.8 ml / min and the column oven temperature is 40 ° C.
  • the acidic soluble soybean protein material of the present invention uses soybean-derived materials.
  • the preparation method of the acid-soluble soybean protein raw material using the raw material derived from soybean is illustrated below.
  • Soy protein material The production of the above-mentioned acidic soluble soybean protein material can be prepared by acid solubilizing the soybean protein material, for example, as follows. First, a soy material that can be extracted with soy protein, including okara (insoluble fiber content), such as defatted soybeans and concentrated soybean protein that has undergone a defatting process. Degreasing is performed by pressing or solvent extraction. Generally, defatted soybeans obtained by low-temperature extraction using n-hexane as an extraction solvent are suitable as a starting material.
  • soy material that can be extracted with soy protein including okara (insoluble fiber content), such as defatted soybeans and concentrated soybean protein that has undergone a defatting process. Degreasing is performed by pressing or solvent extraction. Generally, defatted soybeans obtained by low-temperature extraction using n-hexane as an extraction solvent are suitable as a starting material.
  • soybean protein extracting soluble components from such soybean raw materials with water or hot water to remove insoluble Okara, and adjusting the pH of the defatted soymilk to the vicinity of the isoelectric point of soy protein, protein components that are insoluble components
  • the separated soybean protein obtained by separating and recovering can be used as a soybean protein material in the subsequent steps.
  • a separated soybean protein containing a large amount of 7S globulin or 11S globulin obtained by fractionating separated soybean protein or using soybean rich in specific components can also be used.
  • “soy protein” means a protein derived from soybean or a molecule of a hydrolyzate thereof.
  • non-fat soy, concentrated soy protein and other okaras insoluble fiber content
  • protein components are extracted from the soy raw material and the defatted soy milk or defatted soy milk is isoelectric.
  • the neutralized product of point precipitation (separated soy protein) or a hydrolyzate thereof is expressed and distinguished as “soy protein material”.
  • the acidic soluble soybean protein material of the present invention is preferably a separated soybean protein having a crude protein content of 85% or more. Furthermore, if it is used for foods and drinks that do not require transparency, soybeans and processed soybean products that are not defatted, such as whole soybeans and soybean flour, are used as “soy raw materials”, and the “soy protein material” Can also be used.
  • soybean protein material described above is treated with protease under stronger acidity than before. That is, a soybean protein material aqueous solution having a pH of 3.4 or less, preferably pH 3 or less is prepared, and protease treatment is started.
  • the acid used here is an acid used in foods, and for example, mineral acids such as hydrochloric acid, sulfuric acid, and phosphoric acid are preferable, and two or more acids may be mixed and used.
  • organic acids such as citric acid, malic acid, tartaric acid, lactic acid, gluconic acid, fumaric acid, succinic acid, acetic acid, and oxalic acid can be used in combination with the mineral acid.
  • pH exceeds 3.4, the present invention cannot be achieved. If the pH is too low, it is difficult to react depending on the type of protease, and the ash content of the system increases, which is not preferable. Usually, a pH of 2.5 or higher is preferable.
  • protease used in the present invention is preferably a protein in which amino acids are bound in a chain or an endo-type protease that is an enzyme that hydrolyzes peptide bonds inside the peptide to form several peptides. It is also possible to combine one or more exo-type proteases, which are enzymes that sequentially cleave amino acids and peptides from the amino terminus and carboxy terminus present at the ends of proteins and peptides.
  • endo-type proteases and exo-type proteases can be used as long as they have activity in an environment of pH 3.4 or lower, but those derived from microorganisms such as Aspergillus genus and Rhizopus genus, and those derived from plants. It can be preferably used. Among these, those derived from microorganisms are preferable, those derived from the genus Aspergillus are particularly preferable, and those derived from Aspergillus oryzae are most preferable.
  • the reaction temperature depends on the type of enzyme used, but the reaction is generally performed at 20 ° C or higher and 70 ° C or lower. If it is less than 20 ° C, the protease activity is low, and it may be industrially unfavorable because it is necessary to increase the amount of enzyme added.
  • the degree of degradation in the above-described gel filtration containing SDS is that the content of the fraction having a mass number of 150 kDa or more is 18% or less, and the TCA (0.22M) solubilization rate is 10% or more and 45%. It is essential that: Further, it is preferably 15% or more and 40% or less.
  • the molecular weight will decrease, and the use will be limited due to the occurrence of bitterness, both in terms of physical properties and taste, and if it is less than 10%, astringency cannot be improved, both of which are for the purpose of the present invention. Does not match.
  • the pH of the system is slightly increased by the treatment with acidic protease, the pH at the start of protease treatment is important for the present invention.
  • an acid solubilization treatment In order to make the soybean protein material described above into an acidic soluble soybean protein material, an acid solubilization treatment is required.
  • a method disclosed in WO2002 / 67690 can be used.
  • phytase treatment for removing phytic acid derived from the raw material protein in the solution containing soy protein, and heating the solution containing soy protein at a temperature exceeding 100 ° C in an acidic region from the pH of the isoelectric point of the protein.
  • the method by performing the treatment is preferable because it can efficiently increase the dissolution rate of soy protein under acidity.
  • the origin of the phytase used in the present invention is not particularly limited as long as it is an enzyme or enzyme agent having phytic acid hydrolysis activity.
  • the temperature, time, pH, and amount added can be appropriately set depending on the phytase used.
  • the order in which the phytase treatment and the protease treatment are performed is not particularly limited, and can be performed before the protease treatment, after the protease treatment, or simultaneously. By simultaneously reacting phytase in the protease reaction, the process is simplified and the productivity can be improved.
  • the heat treatment in the present invention is adjusted to a pH lower than the isoelectric point of soybean protein, preferably pH 2.3 to 4.3, and a temperature exceeding 100 ° C., preferably 180 ° C. or less, more preferably 105 ° C. to 160 ° C. Heat with.
  • a pH exceeding the isoelectric point or at a temperature below 100 ° C. acid solubilization does not proceed, and an acid-soluble soybean protein material that meets the present invention cannot be obtained.
  • the temperature exceeds 180 ° C. peptide bonds are partially decomposed, and the quality may be deteriorated.
  • the heating time is preferably 1 second to 1 hour. When heating for less than 1 second, acid solubilization is not sufficiently performed, and when heating for 1 hour or more, there is a possibility of affecting the quality such as flavor.
  • the concentration of the solution containing soy protein is preferably 3 to 30% by weight of solid content, more preferably 8 to 16% by weight of solid content. If the solid content is less than 3% by weight, there is no problem in quality, but the working efficiency tends to decrease. If the solid content exceeds 30% by weight, the viscosity of the solution containing the protein increases remarkably, and the subsequent workability deteriorates. It tends to be. However, when decomposition of the soy protein to be heated is advanced, the effect of thickening is small and the concentration can be increased.
  • the continuous direct heating sterilizer of a steam injection system can be illustrated as a desirable system.
  • This apparatus is a system in which steam is blown into a liquid flowing in a pipe, and can be instantaneously heated to a high temperature exceeding 100 ° C. If the protease and phytase treatments are performed in advance, it is preferable that acidic solubilization treatment and sterilization by heating can be performed simultaneously by subsequent acidic heating.
  • the solution containing the acid-soluble soy protein material that has been subjected to the acid solubilization treatment can be used as it is, but can also be pulverized in order to enhance the convenience of use.
  • the drying method is not particularly limited, and a spray drying apparatus or the like is suitable.
  • the acidic soluble soybean protein material obtained according to the present invention can provide a solution with a pH of 3 to 4.5, which is low in solubility with ordinary separated soybean protein. In this drying, it is preferable to carry out the drying treatment while maintaining the acidity without performing neutralization or the like.
  • acidic food and drink Usually, a protein under weak acidity has a pH near the isoelectric point, the protein aggregates, and it is difficult to obtain an acidic protein food or drink having a good texture without roughness.
  • acidic soluble soybean protein material obtained in the present invention an acidic food and drink having a good taste in the acidic region, a good texture without causing protein precipitation, and a good storage stability can be obtained.
  • acidic foods and drinks include acidic protein beverages. Seasoning at the time of beverage preparation is selected according to tastes such as sugars and fragrances.
  • the protein content is preferably in the range of 1 to 20% by weight, although it depends on the degree of protein intake.
  • an acidic jelly-like food can be prepared by using an acidic soluble soybean protein material obtained by the present invention in combination with an appropriate gelling agent together with sugars, flavors and the like. Moreover, since it has high solubility while being acidic and good in flavor, it is also suitable for use in swallowing foods, acidic whipped creams and the like.
  • ⁇ Crude protein content The nitrogen content was determined based on the Kjeldahl method, multiplied by a coefficient of 6.25 to obtain the crude protein content, and the result was shown in anhydrous conversion.
  • ⁇ NSI water-soluble nitrogen index: Based on the official analysis method BA-11-65 NSI of AOCS (American Oil Chemist Society), the following was performed. Weighed 3.5 g of protein material, added 100 ml of water, stirred with propeller (500 rpm) for 10 minutes, and sampled nitrogen (measured by Kjeldahl method) in the filtrate filtered with No. 5A filter paper. Expressed as a percentage of nitrogen.
  • Dilute acid NSI modified method In the above NSI measurement method, after stirring the propeller, the pH was 3.5 and the dissolved protein was quantified.
  • -TCA (0.22M) solubilization ratio An equal amount of 0.44M trichloroacetic acid (TCA) was added to a 2% by weight aqueous solution of protein material, and the proportion of soluble protein was measured by the Kjeldahl method. As protein degradation proceeds, the TCA solubilization rate increases.
  • ⁇ pH pH was measured at 25 ° C.
  • This skim soymilk is adjusted to pH 4.5 with phosphoric acid and then centrifuged at 2,000 x g using a continuous centrifuge (decanter) to obtain an insoluble fraction (acid precipitation card) and a soluble fraction (whey). It was.
  • the acid precipitation curd was watered to a solid content of 10% by weight to obtain an acid precipitation curd slurry.
  • This acid precipitation curd slurry was also used in the following examples, comparative examples, and experimental examples.
  • the acid precipitation curd slurry having a solid content of 10% by weight was adjusted to pH 3.5 with a phosphoric acid solution and then heated to 50 ° C.
  • 0.1 wt% protease derived from Aspergillus oryzae (“Sumiteam ACP” manufactured by Shin Nippon Chemical Co., Ltd., 1,000 U / g) at a solid content was added and hydrolyzed for 60 minutes. After the reaction, the enzyme was inactivated by heating at 85 ° C. for 20 minutes.
  • This hydrolyzate was adjusted to pH 3.5, phytase equivalent to 8 units per 100 g of solid content (“Phytase Novo” manufactured by Novozymes) was added and allowed to act for 30 minutes. Heated at 140 ° C for 10 seconds in a continuous direct heat sterilizer. This was spray-dried to obtain a powdery acidic soluble soybean protein material (A) (crude protein content 91%).
  • Example 1 Preparation of protease-degrading / acid-soluble soybean protein material at pH 3.3 After adjusting the acid precipitation curd slurry of Comparative Example 1 having a solid content of 10% by weight to pH 3.3 with phosphoric acid, 50 ° C It was heated to become. To this solution, 0.1% by weight of “Sumiteam ACP” per solid content was added and hydrolyzed for 60 minutes. After the reaction, the enzyme was inactivated by heating at 85 ° C. for 20 minutes. This hydrolyzate was adjusted to pH 3.5, “phytase novo” corresponding to 8 units per solid was added, and the enzyme reaction was carried out for 30 minutes. After the reaction, it was heated at 140 ° C. for 10 seconds in a continuous direct heat sterilizer. This was spray-dried to obtain a powdery acidic soluble soybean protein material (B) (crude protein content 90%).
  • B powdery acidic soluble soybean protein material
  • Example 2 Preparation of protease-degrading / acid-soluble soy protein material at pH 3.0 After adjusting the acid precipitation curd slurry of Comparative Example 1 having a solid content of 10% by weight to pH 3.0 with phosphoric acid, 50 ° C It was heated to become. To this solution, 0.1% by weight of “Sumiteam ACP” per solid content was added and hydrolyzed for 60 minutes. After the reaction, the enzyme was inactivated by heating at 85 ° C. for 20 minutes. To this hydrolyzate, “phytase novo” corresponding to 8 units per solid was added, and an enzyme reaction was carried out for 30 minutes. After the reaction, it was heated at 140 ° C. for 10 seconds in a continuous direct heat sterilizer. This was spray-dried to obtain a powdery acidic soluble soybean protein material (C) (crude protein content 90%).
  • C powdery acidic soluble soybean protein material
  • Comparative Example 2 Preparation of undegraded / acid-soluble soy protein material After adjusting the acid precipitation curd slurry having a solid content of 10% by weight of Comparative Example 1 to pH 3.5 with a phosphoric acid solution, “Phytase Novo” equivalent to 8 units was added and allowed to act for 30 minutes. Heated at 140 ° C for 10 seconds in a continuous direct heat sterilizer. These were spray-dried to obtain a powdery acidic soluble soybean protein material (D) (crude protein amount 92%).
  • D powdery acidic soluble soybean protein material
  • Comparative Example 4 Preparation of Neutral Protease Decomposition / Acid Soluble Soy Protein Material After adjusting the acid precipitation curd slurry having a solid content of 10% by weight of Comparative Example 1 to pH 6.5 with dilute sodium hydroxide solution, continuous Heated at 140 ° C for 10 seconds in a direct heat sterilizer. This solution was adjusted to 50 ° C., and 0.1 wt% of protease derived from the genus Bacillus (“Protamex” 1.5 AU (Anson unit) / g, manufactured by Novozyme) per solid content was added, followed by hydrolysis for 60 minutes. After the reaction, the enzyme was inactivated by heating at 85 ° C. for 20 minutes.
  • Example 3 Preparation of protease-degrading / acid-soluble soybean protein material at pH 3.0 2 Preparation was carried out in the same manner as in Example 2 to obtain a powdery acidic soluble soybean protein material (H) (crude protein content 90%). However, “Sumiteam ACP” was added to the solution at a concentration of 0.25 wt% per solid, and hydrolysis was performed for 60 minutes.
  • Sensory evaluation and quality measurement were carried out on the acid-soluble soybean protein materials (A) to (H) (Table 1).
  • Sensory evaluation prepared 5 weight% aqueous solution, and evaluated "astringency” and "bitterness”. For each item, based on “1 point” that you do not feel, “2 points” that you hardly feel, “3 points” that you feel but are uncomfortable, “4 points” that you feel (discomfort), "5 points” that you feel strongly, Sensory evaluation was performed by 20 panelists, and the average value was shown.
  • the sample was dissolved in the eluent so as to be 0.1% by weight as protein, and after passing through a cellulose acetate type membrane filter (DISMIC-13CP 0.20 ⁇ m, manufactured by Advantec) to remove the precipitate, 20 ⁇ l was injected.
  • a cellulose acetate type membrane filter (DISMIC-13CP 0.20 ⁇ m, manufactured by Advantec) to remove the precipitate
  • the HPLC main unit uses equipment manufactured by JASCO Corporation, “PU-2089 Plus” as the liquid feed pump, “UV-2077 Plus” as the detector, “CO-2,065 Plus” as the column oven, and “LC” as the system controller.
  • -Net II / ADC "and” ChromNAV "as analysis software.
  • the chromatogram waveform processing was ChromNAV with a slope sensitivity of 10.00 ⁇ V / sec, a slope width of 0.1 min, a minimum area of 10,000 ⁇ V ⁇ sec, a minimum height of 1000 ⁇ V, a drift of 0.000 ⁇ V / min, and a slope width doubling time of 0.0000 min.
  • Electrophoresis was in accordance with the Laemmli method [Nature, Vol. 227, 680-685 (1970)]. That is, 10 ⁇ L of a 5% by weight aqueous solution of a sample and 90 ⁇ L of Tris-SDS- ⁇ -ME sample treatment solution (manufactured by Cosmo Bio) were mixed and heated at 100 ° C. for 5 minutes, and SDS-Tris-Glycine running buffer (Cosmo Bio 10-20% gradient gel was used, and a constant flow of 25 mA was used for 1 hour. After electrophoresis, the gel was stained with Page Blue 83 (CBB R-250) staining solution (manufactured by Cosmo Bio).
  • CBB R-250 Page Blue 83
  • (F) of neutral degradation has a thinner band observed by electrophoresis than (C), and even though it has a high degree of degradation, the content of fractions of 150 kda or higher by gel filtration is as high as 21.8% ( Compared with Table 1, FIG. 1), (C), astringency was still felt, and also the amount of crude protein was lower than others. From the above examination, it was confirmed that the polymerized product due to SS bond or the like cannot be dissociated by simply performing hydrolysis under the decomposition conditions that do not cause bitterness, and it does not lead to a decrease in astringency.
  • Example 4 Preparation of acidic soluble soybean protein material by decomposition at pH 3.0 using 11S globulin as a raw material 11S globulin from isolated soybean protein was prepared by the production method described in International Publication WO2006 / 129647. To obtain 11S globulin-enriched material. That is, 1 kg of low-denatured defatted soybean was mixed while spraying 100 g of 70 wt% hydrous alcohol, and the outside of the sealed container was heated so that the product temperature of the defatted soybean was 70 ° C. and maintained for 30 minutes.
  • the 11S globulin concentrated material was added with water to a solid content of 10% by weight, adjusted to pH 3.0, 0.1% by weight of “Sumiteam ACP” was added to the solid content, and hydrolysis was performed at 50 ° C. for 60 minutes. After the reaction, the enzyme was inactivated by heating at 85 ° C. for 20 minutes. This hydrolyzate was adjusted to pH 3.5, “phytase Novo” corresponding to 8 units per 100 g of solid content was added, and allowed to act for 30 minutes. Heated at 140 ° C for 10 seconds in a continuous direct heat sterilizer. This was spray-dried to obtain powdered acidic soluble soybean protein material (I) (crude protein content 90%).
  • “Sumiteam ACP” was added to the solid content, and hydrolysis was performed at 50 ° C. for 60 minutes. After the reaction, the enzyme was inactivated by heating at 85 ° C. for 20 minutes. This hydrolyzate was adjusted to pH 3.5, “phytase Novo” corresponding
  • Example 6 Preparation of undegraded acidic soluble soybean protein material using 11S globulin as a raw material
  • the 11S globulin concentrated material having a solid content of 10% by weight described in Example 4 was adjusted to pH 3.0, and 140 ° C. , Heated for 10 seconds and spray-dried to obtain an 11S globulin undegraded product (J) (crude protein amount 92%) subjected to acid solubilization treatment.
  • Example 5 Preparation of acidic soluble soy protein material by decomposition at pH 3.0 using 7S globulin as a raw material 7S globulin from separated soy protein was produced using the production method described in International Publication WO2002 / 28198 And 7S globulin-enriched material was obtained. That is, 10 parts by weight of water is added to 1 part by weight of low-denatured defatted soybeans, extracted with stirring for 1 hour at room temperature while adjusting to pH 7.0 with dilute sodium hydroxide solution, and centrifuged at 1,000 xg for okara and insoluble. Minutes were separated to obtain defatted soymilk.
  • the defatted soymilk was adjusted to pH 4.2 using hydrochloric acid, heated to 40 ° C., cooled, adjusted to pH 5.8 and centrifuged to obtain a soluble fraction.
  • the soluble fraction was adjusted to pH 4.9 with hydrochloric acid and centrifuged at 1,000 ⁇ g to obtain a 7S globulin-enriched material that was a precipitate.
  • the 7S globulin concentrate was hydrated to a solid content of 10% by weight, adjusted to pH 3.0, 0.1% “Sumiteam ACP” was added per solid content, and hydrolysis was performed at 50 ° C. for 60 minutes. After the reaction, the enzyme was inactivated by heating at 85 ° C. for 20 minutes.
  • (L) prepared from a material rich in 7S globulin has low astringency and bitterness, and can be used for acidic foods. From the above, even when using a material rich in 11S globulin and 7S globulin as a raw material, as in Example 2, by starting enzymatic degradation at pH 3.4 or lower, acidic soluble soybean protein to a level that can be used for acidic foods and drinks. The astringency of the material could be reduced.
  • Example 6 Preparation of acidic powder beverage 66.7 parts of powdered acidic soluble soybean protein material (C) prepared in Example 2, 18.6 parts of granulated sugar, 12.4 parts of grapefruit powder juice (made by Yaizu Suisan Chemical Co., Ltd.) , Grapefruit fragrance (manufactured by Takasago Fragrance Co., Ltd.) 2.1 parts and sweetener (Sucralose, Saneigen FFI Co., Ltd.) 0.2 part were mixed well to obtain a protein-containing acidic powder drink. 15 g of this powder was added to 185 g of water and mixed well with a shaker to obtain a beverage. Although this beverage was evaluated by sensory evaluation, it has an appropriate acidity, does not feel the astringent taste peculiar to conventional acidic soluble soybean protein materials, does not feel bitterness, does not feel soybean odor, and feels good over the throat It was excellent in palatability.
  • Example 7 Preparation of acidic jelly-like food 5.0 parts by weight of powdered acid-soluble soybean protein material (C) prepared in Example 2, 8.0 parts of fructose glucose liquid (manufactured by Cornstarch Japan), 5-fold concentrated pineapple Mixing fruit juice (Food Material Co., Ltd.) 2.5 parts, agar (Ina Agar Co., Ltd.) 0.3 parts, pineapple fragrance (Takasago Fragrance Co., Ltd.) 0.2 parts, water 84.0 parts. The mixture was adjusted to pH 3.6, sterilized by heating at 95 ° C., mixed with agar liquid swollen by heating at high temperature, filled in a cheer pack, and gelled by refrigeration overnight to prepare a jelly beverage. The obtained jelly beverage did not feel astringent and had a good texture.
  • Example 8 Preparation of acidic swallow food Powdered acidic soluble soybean protein material prepared in Example 2 (C) 4.3 parts by weight, granulated sugar 12.5 parts by weight, fermented lactate dolomite (Meiji Dairies) 1.25 parts by weight, Vitamin fortifier 0.125 parts by weight, tartaric acid 0.1 parts by weight, mineral yeast preparation “Mix Mineral Yeast C” 0.1 part by weight, “Mix Mineral Yeast M” 0.025 part by weight (manufactured by LS Corporation), stevia preparation 0.06 part by weight The powder was mixed and dispersed and dissolved in 46.04 parts by weight of water. A high-speed emulsification disperser was used for dissolution.
  • a jelly-like swallowed food (pH 3.90). It contains protein, lipid, and sugar, which are the three major nutrients, and can consume 1.2kcal in 1g, and other nutrients necessary for the body: calcium, magnesium, trace element minerals, vitamins, and dietary fiber.
  • Example 9 Preparation of acidic whipped cream To 2.0 parts by weight of powdered acidic soluble soybean protein material (C) prepared in Example 2, 0.2 parts by weight of sucrose fatty acid ester S-570 (manufactured by Mitsubishi Chemical Foods) Then, 52.6 parts by weight of water was added and dissolved, and an aqueous phase was prepared at a pH of 3.5. Separately, 23 parts by weight of a middle melting point fraction of palm oil, 22 parts by weight of hardened coconut oil, and 0.2 parts by weight of soybean lecithin (manufactured by Sakai Oil Co., Ltd.) were mixed to prepare an oil phase. The aqueous phase and the oil phase were mixed, stirred with a homomixer at 70 ° C.
  • the present invention it is possible to produce a protein material that has little astringency and bitterness, excellent flavor, and satisfies the physicochemical properties such as low viscosity, high solubility, and high stability required for acidic foods and drinks. .
  • this acidic soluble soybean protein material protein-containing acidic foods and drinks that have a better flavor than before can be produced.

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Abstract

Disclosed is an acid-soluble soybean protein material which can be used in an acidic food or beverage, has reduced astringency and reduced bitterness, and has a excellent flavor. The acid-soluble soybean protein material contains a fraction that is detected at 220 nm in gel filtration using a buffer solution containing 1% by weight of SDS (pH 7.0) and has a mass number of 150 kDa or more in an amount of 18% or less, and has a TCA (0.22 M) solubilization rate of 45% or less. Also disclosed is a process for producing an acid-soluble soybean protein material by carrying out a treatment with a phytase and a heating treatment at a temperature higher than 100°C under acidic conditions. In the process, a treatment with a protease is initiated at pH 3.4 or lower, whereby an acid-soluble soybean protein material having the above-mentioned properties can be produced.

Description

酸性可溶大豆たん白素材およびその製造方法Acid-soluble soybean protein material and method for producing the same
　本発明は、例えば酸性飲食物に有用な酸性可溶大豆たん白素材に関する。 The present invention relates to an acid-soluble soybean protein material useful for, for example, acidic foods and drinks.
　酸性の飲食物は、その爽やかな風味を特徴としており、果汁飲料等の酸性飲料やゼリー、並びにヨーグルト類が代表的である。従来、透明性の高いたん白飲食物を調製しようとした場合、酸性条件下で溶解性の高い、乳ホエーの主成分である乳清たん白が専ら用いられてきた。
　大豆から分離して得た大豆たん白素材である分離大豆たん白は、pH3付近よりその等電点であるpH4.5前後のpH域においては、溶解性が非常に低い。そこで、大豆蛋白質を含む分離大豆たん白の酸性水への溶解性を高めるために、種々の試みが行われている。特許文献１は、大豆蛋白質をpH2.0～4.2の酸性下で、250～320°F（121～160℃）に加熱することで、酸性での大豆蛋白質の溶解性を上げている。また、特許文献２は、分離大豆たん白をフィターゼ処理し、続けて酸性下で高温加熱処理を行う等の処理により、酸性域での大豆たん白素材の溶解性を大きく向上させている。 Acidic foods and drinks are characterized by their refreshing flavor, and are typically acidic beverages such as fruit juices, jelly, and yogurts. Conventionally, when preparing highly transparent protein foods and drinks, whey protein, which is a main component of milk whey and has high solubility under acidic conditions, has been exclusively used.
Separated soy protein, which is a soy protein material obtained by separating from soy, has a very low solubility in the pH range around pH 4.5, which is its isoelectric point from around pH 3. Therefore, various attempts have been made to increase the solubility of soy protein containing soy protein in acidic water. In Patent Document 1, the soy protein is heated to 250 to 320 ° F. (121 to 160 ° C.) under an acid pH of 2.0 to 4.2 to increase the solubility of the soy protein in the acid state. Patent Document 2 greatly improves the solubility of the soy protein material in the acidic region by a treatment such as phytase treatment of the separated soybean protein followed by high-temperature heat treatment under acidity.
　乳清たん白や酸性可溶大豆たん白素材などの、pH3.5で溶解するこれらたん白素材の酸性水溶液は、摂取時に口中で渋味を感じる問題がある（非特許文献１）。この渋みは口中で蛋白質が等電点沈殿することにより誘発され、蛋白質を低分子化することで回避が可能とされている。しかしながら、弱酸性下での蛋白質の加水分解は渋味を抑える反面、苦味が発生する問題があり、この渋味および苦味の双方を飲食物に用いられるレベルまで低下させることができなかった。 Acidic aqueous solutions of these protein materials that dissolve at pH 3.5, such as whey protein and acid-soluble soy protein materials, have the problem of feeling astringency in the mouth when ingested (Non-patent Document 1). This astringency is induced by isoelectric precipitation of the protein in the mouth, and can be avoided by reducing the molecular weight of the protein. However, protein hydrolysis under weak acidity reduces astringency, but has a problem of bitterness. Both astringency and bitterness could not be reduced to a level used for food and drink.
　そこで苦味発生を抑えつつ渋味を低減させるため、中性下で加水分解を行った後に弱酸性に調整し、酸性可溶化処理を行う方法も検討された（特許文献３）。この中性分解法では、苦味はほとんど発生せず、渋味もかなり低下したものが調製できたが、尚一層の改善が望まれている。渋味，苦味が低減し、飲料またはゼリー状食品等の酸性飲食物に利用できるレベルに達した、酸性可溶大豆たん白素材が切望されている。 Therefore, in order to reduce the astringency while suppressing the occurrence of bitterness, a method of performing acid solubilization treatment by adjusting to weak acidity after performing hydrolysis under neutrality was also studied (Patent Document 3). In this neutral decomposition method, a bitterness was hardly generated and astringency was considerably reduced, but further improvement is desired. An acidic soluble soybean protein material that has reduced astringency and bitterness and has reached a level that can be used for acidic foods such as beverages or jelly-like foods is desired.
特公昭53-19669公報Japanese Patent Publication No. 53-19669 WO02/067690パンフレットWO02 / 067690 Brochure WO2008/136326パンフレットWO2008 / 136326 Brochure
　本発明の目的は、酸性飲食物に用いるための、渋味，苦味が低減され風味に優れた、酸性可溶大豆たん白素材を提供することにある。 An object of the present invention is to provide an acidic-soluble soybean protein material that is reduced in astringency and bitterness and excellent in flavor for use in acidic foods and drinks.
　本発明者らは上記の問題を解決すべく鋭意研究の結果、酸性可溶大豆たん白素材の調製に際して、大豆蛋白質含有溶液を一定の分解度までプロテアーゼ分解するに際して、これまで行われていなかったpH3.4以下で反応を開始することで、渋味および苦味が大きく低下することを見出すと共に、このものは後述するゲル濾過での150kDa以上の画分の含量が有意に低いことを発見し、この発明を完成させた。
　すなわち本発明は、
（１）１重量%SDS含有緩衝液（pH7.0）を用いたゲル濾過において、220nmで検出される、質量数150kDa以上の画分の含量が18%以下で且つTCA（0.22M）可溶化率が45%以下の、酸性可溶大豆たん白素材。
（２）pH3.4以下でプロテアーゼ処理を開始し、TCA（0.22M）可溶化率を10%以上，45%以下とすることを特徴とする、酸性可溶大豆たん白素材の製造方法。
（３）酸性可溶大豆たん白素材の製造が、フィターゼ処理および100℃を超える酸性加熱処理によるものである、（２）に記載の酸性可溶大豆たん白素材の製造方法。
（４）請求項１に記載の酸性可溶大豆たん白素材を用いた、酸性飲食物。 As a result of diligent research to solve the above-mentioned problems, the present inventors have not so far carried out protease degradation of a soy protein-containing solution to a certain degree of degradation when preparing an acidic soluble soybean protein material. It was found that by starting the reaction at pH 3.4 or less, astringency and bitterness are greatly reduced, and this discovered that the content of fractions of 150 kDa or more in gel filtration described later was significantly low, This invention was completed.
That is, the present invention
(1) In gel filtration using 1 wt% SDS-containing buffer solution (pH 7.0), the content of the fraction with mass number of 150 kDa or more detected at 220 nm is 18% or less and TCA (0.22M) is solubilized. Acid soluble soy protein material with a rate of 45% or less.
(2) A method for producing an acidic soluble soybean protein material, characterized in that protease treatment is started at pH 3.4 or less, and the TCA (0.22M) solubilization rate is 10% or more and 45% or less.
(3) The method for producing an acid-soluble soybean protein material according to (2), wherein the production of the acid-soluble soybean protein material is based on a phytase treatment and an acidic heat treatment exceeding 100 ° C.
(4) Acidic food and drink using the acidic soluble soybean protein material according to claim 1.
　本発明により、渋味も苦味も少なく風味に優れた、酸性飲食物に好適な酸性可溶大豆たん白素材を得ることができる。 According to the present invention, it is possible to obtain an acidic-soluble soybean protein material suitable for acidic foods and drinks, having little astringency and bitterness and excellent flavor.
ゲル濾過による比較　本発明品（C）は、従来品（F）と比較し、150kDa以上の画分が非常に少ないことを示したゲル濾過クロマトグラムである。Comparison by gel filtration The product (C) of the present invention is a gel filtration chromatogram showing that the fraction of 150 kDa or more is very small compared to the conventional product (F). 電気泳動による比較　本発明品（C）のみならず、従来品（F）も、7Sグロブリン，11Sグロブリンのバンドが残存していないことを示した、SDSポリアクリルアミドゲル電気泳動パターンの図である。Comparison by electrophoresis FIG. 7 is an SDS polyacrylamide gel electrophoresis pattern showing that the 7S globulin and 11S globulin bands do not remain in the product (C) of the present invention as well as the conventional product (F).
（酸性可溶大豆たん白素材）
　以下、本発明を具体的に説明する。本明細書に於いては、酸性可溶大豆たん白素材を、大豆原料より抽出しオカラを分離した蛋白質に富んだ画分であって、希酸NSI変法が90%以上のものと定義する。更に本発明の１形態は、酸性可溶大豆たん白素材であって、後述する分子量分布を示すと共に、TCA（0.22M）可溶化率が45%以下のものであり、40%以下であるとさらに好ましい。TCA（0.22M）可溶化率が45%を超えると、苦みが多く発生する場合がある。また希酸NSIが90%未満では、ザラツキ等が発生し、酸性飲食物用たん白素材としては使用が困難である。 (Acid-soluble soy protein material)
The present invention will be specifically described below. In the present specification, an acidic soluble soybean protein material is a protein-rich fraction extracted from soybean raw material and separated from okara, and is defined as a dilute acid NSI modified method of 90% or more. . Furthermore, one form of the present invention is an acidic soluble soybean protein material, which exhibits a molecular weight distribution described later, has a TCA (0.22M) solubilization rate of 45% or less, and 40% or less. Further preferred. When the TCA (0.22M) solubilization rate exceeds 45%, a lot of bitterness may occur. Further, when the dilute acid NSI is less than 90%, roughness and the like occur, and it is difficult to use as a protein material for acidic foods and drinks.
（分子量分布）
　本発明の１形態は、１重量%SDS含有緩衝液（pH7.0）を用いたゲル濾過において、220nmで検出される、質量数150kDa以上の画分の含量が18%以下、好ましくは14%以下の酸性可溶大豆たん白素材である。質量数150kDa以上の画分の含量が18%を超える酸性可溶大豆たん白素材では、不快なレベルの渋味発生が大きく、酸性飲食物に用いることができない。一方、18%以下では不快なレベル以下に渋味が低減し、14%以下ではほぼ渋味が感じられなくなる。以上の条件に合致したたん白素材のみが、本発明であり、酸性飲食物に好適である。 (Molecular weight distribution)
In one embodiment of the present invention, the content of a fraction having a mass number of 150 kDa or more detected at 220 nm in gel filtration using a 1 wt% SDS-containing buffer (pH 7.0) is 18% or less, preferably 14%. The following acidic soluble soybean protein materials. An acidic soluble soybean protein material having a mass fraction of more than 150 kDa with a content of more than 18% has a large unpleasant astringency and cannot be used for acidic foods. On the other hand, if it is 18% or less, the astringency is reduced to an unpleasant level or less, and if it is 14% or less, it is almost impossible to feel astringency. Only protein materials that meet the above conditions are the present invention and are suitable for acidic foods and drinks.
　このゲル濾過は以下の様に行う事ができる。すなわち、「TSK gel G3000SWXL 」(φ7.8mm×300mm)（東ソー社製）カラムを用い、溶離液には１%SDSおよび1.17%NaClを含んだ50mMリン酸緩衝液（pH7.0）を用い、流速を0.8ml/min，カラムオーブン温度40℃とする。 This gel filtration can be performed as follows. That is, a column of “TSK gel G3000SWXL” (φ7.8mm × 300mm) (manufactured by Tosoh Corporation) was used, and a 50 mM phosphate buffer (pH 7.0) containing 1% SDS and 1.17% NaCl was used as the eluent. The flow rate is 0.8 ml / min and the column oven temperature is 40 ° C.
　測定波長220nmにて得られるゲル濾過の結果から、分子量マーカーであるγ-globlin（分子量150kDa）の溶出時間より早い溶出時間のピーク面積を測定し、サンプルのピーク面積全体に対する割合として、質量数150kDa以上の画分の含量を求める。 From the results of gel filtration obtained at a measurement wavelength of 220 nm, the peak area of the elution time earlier than the elution time of the molecular weight marker γ-globlin (molecular weight 150 kDa) is measured, and the mass number is 150 kDa as a percentage of the total peak area of the sample. Obtain the content of the above fractions.
　本発明の酸性可溶大豆たん白素材は大豆由来の原料を使用する。以下に大豆由来の原料を用いた、酸性可溶大豆たん白素材の調製法を例示する。 The acidic soluble soybean protein material of the present invention uses soybean-derived materials. The preparation method of the acid-soluble soybean protein raw material using the raw material derived from soybean is illustrated below.
（大豆たん白素材）
　上記の酸性可溶大豆たん白素材の製造は、大豆たん白素材を酸性可溶化することで調製でき、例えば以下のように行う。まず、脱脂工程を経た脱脂大豆，濃縮大豆たん白等で、オカラ（不溶性繊維分）を含んだ、大豆蛋白質が抽出可能なものを大豆原料とする。脱脂には圧搾や溶剤抽出を行うが、一般的には、n-ヘキサンを抽出溶剤として低温抽出を行った脱脂大豆が出発原料として適当である。このような大豆原料から水または湯で可溶成分を抽出し、不溶性のオカラを除去した脱脂豆乳や、脱脂豆乳を大豆蛋白質の等電点付近のpHに調整して、不溶成分である蛋白質成分を分離回収した分離大豆たん白を、大豆たん白素材として以降の工程に用いることができる。また本発明には、分離大豆たん白を分画することや、特定の成分に富んだ大豆を用いること等により得られる、７Sグロブリンまたは11Sグロブリンを多く含む分離大豆たん白も使用できる。尚、本明細書において、「大豆蛋白質」とは大豆に由来する蛋白質またはその加水分解物の分子を意味する。対して、脱脂大豆，濃縮大豆たん白等のオカラ（不溶性繊維分）を分離していないものを「大豆原料」、大豆原料から蛋白質成分を抽出しオカラを分離した脱脂豆乳若しくは脱脂豆乳の等電点沈殿の中和物（分離大豆たん白）、またはそれらの加水分解物等を「大豆たん白素材」と表現し区別している。本発明である酸性可溶大豆たん白素材は、粗蛋白分が85%以上ある分離大豆たん白であることが好ましい。
　更に、透明性を求めない飲食物への用途であれば、丸大豆，大豆粉等の、脱脂していない大豆や大豆加工品を「大豆原料」として用い、そこから得られる「大豆たん白素材」を使用することも可能である。 (Soy protein material)
The production of the above-mentioned acidic soluble soybean protein material can be prepared by acid solubilizing the soybean protein material, for example, as follows. First, a soy material that can be extracted with soy protein, including okara (insoluble fiber content), such as defatted soybeans and concentrated soybean protein that has undergone a defatting process. Degreasing is performed by pressing or solvent extraction. Generally, defatted soybeans obtained by low-temperature extraction using n-hexane as an extraction solvent are suitable as a starting material. Extracting soluble components from such soybean raw materials with water or hot water to remove insoluble Okara, and adjusting the pH of the defatted soymilk to the vicinity of the isoelectric point of soy protein, protein components that are insoluble components The separated soybean protein obtained by separating and recovering can be used as a soybean protein material in the subsequent steps. In the present invention, a separated soybean protein containing a large amount of 7S globulin or 11S globulin obtained by fractionating separated soybean protein or using soybean rich in specific components can also be used. In the present specification, “soy protein” means a protein derived from soybean or a molecule of a hydrolyzate thereof. In contrast, non-fat soy, concentrated soy protein and other okaras (insoluble fiber content) are separated from “soy raw material”, and protein components are extracted from the soy raw material and the defatted soy milk or defatted soy milk is isoelectric. The neutralized product of point precipitation (separated soy protein) or a hydrolyzate thereof is expressed and distinguished as “soy protein material”. The acidic soluble soybean protein material of the present invention is preferably a separated soybean protein having a crude protein content of 85% or more.
Furthermore, if it is used for foods and drinks that do not require transparency, soybeans and processed soybean products that are not defatted, such as whole soybeans and soybean flour, are used as “soy raw materials”, and the “soy protein material” Can also be used.
（酸添加処理）
　続けて、前述した大豆たん白素材を、従来より強い酸性下でプロテアーゼ処理することが、本発明品を調製する上で重要である。すなわち、pH3.4以下、好ましくはpH３以下の大豆たん白素材水溶液を調製し、プロテアーゼ処理を開始する。ここで用いる酸は食品に用いる酸であって、例えば、塩酸，硫酸，リン酸等の鉱酸が好ましく、２種以上の酸を混合して用いても良い。また、クエン酸，リンゴ酸，酒石酸，乳酸，グルコン酸，フマル酸，コハク酸，酢酸，シュウ酸などの有機酸を、鉱酸に併用することもできる。pHが3.4を超えると本発明は達成できず、またpHが低すぎると、プロテアーゼの種類によっては反応しにくくなる上に、系の灰分が増えて好ましくない。通常はpH2.5以上が好ましい。 (Acid addition treatment)
Subsequently, it is important in preparing the product of the present invention that the soybean protein material described above is treated with protease under stronger acidity than before. That is, a soybean protein material aqueous solution having a pH of 3.4 or less, preferably pH 3 or less is prepared, and protease treatment is started. The acid used here is an acid used in foods, and for example, mineral acids such as hydrochloric acid, sulfuric acid, and phosphoric acid are preferable, and two or more acids may be mixed and used. In addition, organic acids such as citric acid, malic acid, tartaric acid, lactic acid, gluconic acid, fumaric acid, succinic acid, acetic acid, and oxalic acid can be used in combination with the mineral acid. If the pH exceeds 3.4, the present invention cannot be achieved. If the pH is too low, it is difficult to react depending on the type of protease, and the ash content of the system increases, which is not preferable. Usually, a pH of 2.5 or higher is preferable.
（プロテアーゼ処理）
　この溶液にプロテアーゼを加え反応させる。本発明で用いるプロテアーゼとしては、アミノ酸が鎖状に結合する蛋白質やペプチド内部のペプチド結合を加水分解し、いくつかのペプチドとする酵素であるエンド型プロテアーゼが好適である。また蛋白質やペプチドの端に存在するアミノ末端及びカルボキシ末端からアミノ酸やペプチドなどを順に切断する酵素であるエキソ型プロテアーゼを1種類以上組み合わせることも可能である。これらエンド型プロテアーゼおよびエキソ型プロテアーゼの種類は、pH3.4以下の環境で活性を持つものであれば使用できるが、Aspergillus属やRhizopus属などの微生物に由来するものや、植物に由来するものが好ましく使用できる。中でも、微生物に由来するものが好ましく、Aspergillus属に由来するものが特に好ましく、Aspergillus oryzaeに由来するものが最も好ましい。Aspergillus属に由来するプロテアーゼとして、「スミチームACP」（新日本化学社製），「プロテアーゼM（アマノ）G」（天野エンザイム社製）（以上、Aspergillus oryzae由来）、「オリエンターゼ20A」（エイチビィアイ社製），「デナプシン2P」（ナガセケムテック社製）（以上、Aspergillus niger由来）等を挙げることができる。 (Protease treatment)
Protease is added to this solution and reacted. The protease used in the present invention is preferably a protein in which amino acids are bound in a chain or an endo-type protease that is an enzyme that hydrolyzes peptide bonds inside the peptide to form several peptides. It is also possible to combine one or more exo-type proteases, which are enzymes that sequentially cleave amino acids and peptides from the amino terminus and carboxy terminus present at the ends of proteins and peptides. These endo-type proteases and exo-type proteases can be used as long as they have activity in an environment of pH 3.4 or lower, but those derived from microorganisms such as Aspergillus genus and Rhizopus genus, and those derived from plants. It can be preferably used. Among these, those derived from microorganisms are preferable, those derived from the genus Aspergillus are particularly preferable, and those derived from Aspergillus oryzae are most preferable. As proteases derived from the genus Aspergillus, "Sumiteam ACP" (manufactured by Shinnippon Chemical Co., Ltd.), "Protease M (Amano) G" (manufactured by Amano Enzyme) (above, derived from Aspergillus oryzae), "Orientase 20A" (HTV Corporation) And "Denapsin 2P" (manufactured by Nagase Chemtech) (from Aspergillus niger).
　反応温度は、使用する酵素の種類にもよるが、概ね20℃以上、70℃以下で反応させる。20℃未満ではプロテアーゼ活性が低く酵素添加量を増加させる必要から工業的に好ましくない場合があり、また70℃を超える場合、プロテアーゼの種類によっては熱変性により失活する場合がある。分解の程度は、前述したSDSを含むゲル濾過に於いて、質量数150kDa以上の画分の含量が18%以下であり、且つ、TCA（0.22M）可溶化率として、10%以上，45%以下であることが必須である。また、15%以上，40%以下であることが好ましい。45%を超えると低分子化が進行し、物性的にも味的にも、特に苦みの発生により使用が制限され、また10%を下回ると渋みが改善できず、いずれも本発明の目的に合致しない。尚、酸性下のプロテアーゼ処理により、系のpHは多少上昇するが、本発明にはプロテアーゼ処理開始時のpHが重要である。 The reaction temperature depends on the type of enzyme used, but the reaction is generally performed at 20 ° C or higher and 70 ° C or lower. If it is less than 20 ° C, the protease activity is low, and it may be industrially unfavorable because it is necessary to increase the amount of enzyme added. The degree of degradation in the above-described gel filtration containing SDS is that the content of the fraction having a mass number of 150 kDa or more is 18% or less, and the TCA (0.22M) solubilization rate is 10% or more and 45%. It is essential that: Further, it is preferably 15% or more and 40% or less. If it exceeds 45%, the molecular weight will decrease, and the use will be limited due to the occurrence of bitterness, both in terms of physical properties and taste, and if it is less than 10%, astringency cannot be improved, both of which are for the purpose of the present invention. Does not match. Although the pH of the system is slightly increased by the treatment with acidic protease, the pH at the start of protease treatment is important for the present invention.
（酸性可溶化処理）
　上記の大豆たん白素材を酸性可溶大豆たん白素材とするために、酸性可溶化処理が必要となる。本発明で云う酸性可溶化処理は、例えばWO2002/67690号公報に公開されている方法を用いることができる。中でも、大豆蛋白質を含む溶液中の原料蛋白質由来のフィチン酸を除去するためのフィターゼ処理と、大豆蛋白質を含む溶液を当該蛋白質の等電点のpHより酸性域で、100℃を超える温度で加熱する処理を行うことによる方法は、大豆蛋白質の酸性下における溶解率を効率的に高めることができ好ましい。 (Acid solubilization treatment)
In order to make the soybean protein material described above into an acidic soluble soybean protein material, an acid solubilization treatment is required. For the acid solubilization treatment in the present invention, for example, a method disclosed in WO2002 / 67690 can be used. Above all, phytase treatment for removing phytic acid derived from the raw material protein in the solution containing soy protein, and heating the solution containing soy protein at a temperature exceeding 100 ° C in an acidic region from the pH of the isoelectric point of the protein. The method by performing the treatment is preferable because it can efficiently increase the dissolution rate of soy protein under acidity.
（フィターゼ処理）
　本発明に使用するフィターゼは、フィチン酸加水分解活性を有する酵素または酵素剤であれば特に起源は限定されない。温度，時間，pH，添加量は、使用するフィターゼにより適宜設定できる。フィターゼ処理と上記プロテアーゼ処理を行う順序は特に限定されず、プロテアーゼ処理前に行うことも、プロテアーゼ処理後に行うことも、同時に行うことも可能である。上記プロテアーゼの反応に於いてフィターゼを同時に反応させることで、工程が簡略となり、生産性を向上できる。 (Phytase treatment)
The origin of the phytase used in the present invention is not particularly limited as long as it is an enzyme or enzyme agent having phytic acid hydrolysis activity. The temperature, time, pH, and amount added can be appropriately set depending on the phytase used. The order in which the phytase treatment and the protease treatment are performed is not particularly limited, and can be performed before the protease treatment, after the protease treatment, or simultaneously. By simultaneously reacting phytase in the protease reaction, the process is simplified and the productivity can be improved.
（加熱処理）
　本発明における加熱処理は、大豆蛋白質の等電点未満のpH、好ましくはpH2.3～4.3に調整し、且つ、100℃を超える温度、好ましくは180℃以下、更に好ましくは105℃～160℃で加熱する。等電点を超えるpHや100℃未満の温度では、酸性可溶化が進行せず、本発明に合致する酸性可溶大豆たん白素材を得ることができない。温度が180℃を超える場合は、一部にペプチド結合の分解が生じ、品質が劣化する可能性がある。pHが低すぎる場合は、使用する酸の量が著しく増加し、得られる物の風味としても作業性としても好ましくない。加熱時間は1秒間～１時間が好ましい。１秒間未満の加熱では酸性可溶化が十分に行われず、また１時間以上の加熱を行った場合、風味等の品質への影響を及ぼす恐れがある。大豆蛋白質を含む溶液の濃度は、好ましくは固形分３重量%～30重量%、更に好ましくは固形分８重量%～16重量%である。固形分が３重量%未満の場合、品質は問題ないが作業効率が低下する傾向にあり、固形分30重量%を超える場合、蛋白質を含む溶液の粘度が著しく上昇し、その後の作業性を悪化させる傾向にある。ただし、加熱する大豆蛋白質の分解が進んでいる場合は、増粘の影響が小さく、濃度を高くすることも可能である。 (Heat treatment)
The heat treatment in the present invention is adjusted to a pH lower than the isoelectric point of soybean protein, preferably pH 2.3 to 4.3, and a temperature exceeding 100 ° C., preferably 180 ° C. or less, more preferably 105 ° C. to 160 ° C. Heat with. At a pH exceeding the isoelectric point or at a temperature below 100 ° C., acid solubilization does not proceed, and an acid-soluble soybean protein material that meets the present invention cannot be obtained. When the temperature exceeds 180 ° C., peptide bonds are partially decomposed, and the quality may be deteriorated. If the pH is too low, the amount of acid to be used is remarkably increased, which is not preferable in terms of flavor and workability of the resulting product. The heating time is preferably 1 second to 1 hour. When heating for less than 1 second, acid solubilization is not sufficiently performed, and when heating for 1 hour or more, there is a possibility of affecting the quality such as flavor. The concentration of the solution containing soy protein is preferably 3 to 30% by weight of solid content, more preferably 8 to 16% by weight of solid content. If the solid content is less than 3% by weight, there is no problem in quality, but the working efficiency tends to decrease. If the solid content exceeds 30% by weight, the viscosity of the solution containing the protein increases remarkably, and the subsequent workability deteriorates. It tends to be. However, when decomposition of the soy protein to be heated is advanced, the effect of thickening is small and the concentration can be increased.
　加熱方法は問わないが、望ましい方式としてスチームインジェクション方式の連続式直接加熱殺菌装置が例示できる。この装置は管内に流れる液体に蒸気を吹き込む方式で、瞬間的に100℃を超える高温に加熱することができる。上記のプロテアーゼ，フィターゼ処理を予め行っていれば、その後の酸性加熱により、加熱による酸性可溶化処理と殺菌を同時に行えるので好ましい。 Although a heating method is not ask | required, the continuous direct heating sterilizer of a steam injection system can be illustrated as a desirable system. This apparatus is a system in which steam is blown into a liquid flowing in a pipe, and can be instantaneously heated to a high temperature exceeding 100 ° C. If the protease and phytase treatments are performed in advance, it is preferable that acidic solubilization treatment and sterilization by heating can be performed simultaneously by subsequent acidic heating.
（乾燥）
　以上の酸性可溶化処理を行った酸性可溶大豆たん白素材を含む溶液は、溶液のままで使うことももちろん可能であるが、利用上の利便性を高めるために、粉末化することもできる。乾燥の方法は特に限定されず、噴霧乾燥装置などが好適である。本発明によって得られた酸性可溶大豆たん白素材は、通常の分離大豆たん白では溶解性の低い、pH3～4.5の溶液を得ることができる。この乾燥の際は、中和等を行うことなく、酸性のまま乾燥処理を行うことが好ましい。 (Dry)
The solution containing the acid-soluble soy protein material that has been subjected to the acid solubilization treatment can be used as it is, but can also be pulverized in order to enhance the convenience of use. . The drying method is not particularly limited, and a spray drying apparatus or the like is suitable. The acidic soluble soybean protein material obtained according to the present invention can provide a solution with a pH of 3 to 4.5, which is low in solubility with ordinary separated soybean protein. In this drying, it is preferable to carry out the drying treatment while maintaining the acidity without performing neutralization or the like.
（酸性飲食物）
　通常、弱酸性下の蛋白質は、そのpHが等電点付近となり、蛋白質は凝集し、ざらつきのない良好な食感の酸性たん白飲食物は得がたい。本発明で得られる酸性可溶大豆たん白素材を用いることにより、酸性域で風味が良く、蛋白の沈殿を起こさず食感が良好で、保存安定性の良い酸性飲食物が得られる。酸性飲食物の例としては酸性たん白飲料を挙げることができる。飲料作製時の味付けは、糖類，香料等嗜好に合わせて選ばれる。蛋白質の含有量は、蛋白質摂取の必要度にもよるが、１～20重量%の濃度が好ましい範囲である。また、本発明により得られる酸性可溶大豆たん白素材を用い、糖類，香料等と共に適当なゲル化剤と併用することにより、酸性ゼリー状食品も作成できる。
　また、酸性で風味が良いままに高い溶解性を持つことから、嚥下食や酸性ホイップクリーム等への使用も好適である。 (Acid food and drink)
Usually, a protein under weak acidity has a pH near the isoelectric point, the protein aggregates, and it is difficult to obtain an acidic protein food or drink having a good texture without roughness. By using the acidic soluble soybean protein material obtained in the present invention, an acidic food and drink having a good taste in the acidic region, a good texture without causing protein precipitation, and a good storage stability can be obtained. Examples of acidic foods and drinks include acidic protein beverages. Seasoning at the time of beverage preparation is selected according to tastes such as sugars and fragrances. The protein content is preferably in the range of 1 to 20% by weight, although it depends on the degree of protein intake. In addition, an acidic jelly-like food can be prepared by using an acidic soluble soybean protein material obtained by the present invention in combination with an appropriate gelling agent together with sugars, flavors and the like.
Moreover, since it has high solubility while being acidic and good in flavor, it is also suitable for use in swallowing foods, acidic whipped creams and the like.
　本発明に用いた分析方法を以下に記載する。
●粗蛋白量：ケルダール法に基づき窒素含量を求め、係数6.25を乗じて粗蛋白量とし、無水換算して示した。
●NSI（水溶性窒素指数）：AOCS（American Oil Chemistユs Society）の公式分析法BA-11-65 NSIに基づき、以下のように行った。たん白素材3.5gを秤量し、100mlの水を加えてプロペラ撹拌（500rpm）を10分間して、No.5A濾紙で濾過した濾液中の窒素（ケルダール法による測定）をサンプリングしたたん白素材中の窒素に対する百分率で表した。
●希酸NSI変法：上記NSIの測定方法に於いて、プロペラ撹拌後にpHを3.5とし、溶解した蛋白質を定量した。
●TCA（0.22M）可溶化率：たん白素材の２重量%水溶液に、0.44Mトリクロロ酢酸（TCA）を等量加え、可溶性蛋白質の割合をケルダール法により測定した。蛋白質の分解が進行すると、TCA可溶化率は上昇する。
●pH：pHは25℃にて測定した。 The analysis method used in the present invention is described below.
● Crude protein content: The nitrogen content was determined based on the Kjeldahl method, multiplied by a coefficient of 6.25 to obtain the crude protein content, and the result was shown in anhydrous conversion.
● NSI (water-soluble nitrogen index): Based on the official analysis method BA-11-65 NSI of AOCS (American Oil Chemist Society), the following was performed. Weighed 3.5 g of protein material, added 100 ml of water, stirred with propeller (500 rpm) for 10 minutes, and sampled nitrogen (measured by Kjeldahl method) in the filtrate filtered with No. 5A filter paper. Expressed as a percentage of nitrogen.
Dilute acid NSI modified method: In the above NSI measurement method, after stirring the propeller, the pH was 3.5 and the dissolved protein was quantified.
-TCA (0.22M) solubilization ratio: An equal amount of 0.44M trichloroacetic acid (TCA) was added to a 2% by weight aqueous solution of protein material, and the proportion of soluble protein was measured by the Kjeldahl method. As protein degradation proceeds, the TCA solubilization rate increases.
● pH: pH was measured at 25 ° C.
　以下に実施例を記載する。尚、特に記載のない限り、以下の部や%は全て重量部，重量%とした。 Examples are described below. Unless otherwise specified, the following parts and% are all parts by weight and% by weight.
（比較例１）pH3.5でのプロテアーゼ分解・酸性可溶大豆たん白素材の調製
　大豆を圧扁し、n-ヘキサンを抽出溶媒として油を抽出分離除去して得られた低変性脱脂大豆（NSI:91）１重量部に７重量部の水を加え、希水酸化ナトリウム溶液でpH７に調整しつつ室温で１時間撹拌しながら抽出後、4,000×gで遠心分離しオカラおよび不溶分を分離し、脱脂豆乳を得た。この脱脂豆乳をリン酸にてpH4.5に調整後、連続式遠心分離機（デカンター）を用い2,000×gで遠心分離し、不溶性画分（酸沈殿カード）および可溶性画分（ホエー）を得た。酸沈殿カードを固形分10重量%になるように加水し酸沈殿カードスラリーを得た。この酸沈殿カードスラリーは以降の実施例，比較例，実験例にも用いた。 (Comparative Example 1) Preparation of protease-degraded / acid-soluble soybean protein material at pH 3.5 Low-denatured defatted soybean obtained by pressing soybean, extracting and removing oil using n-hexane as an extraction solvent ( NSI: 91) Add 7 parts by weight of water to 1 part by weight, extract with stirring at room temperature for 1 hour while adjusting to pH 7 with dilute sodium hydroxide solution, then centrifuge at 4,000 xg to separate okara and insoluble matter. And defatted soy milk was obtained. This skim soymilk is adjusted to pH 4.5 with phosphoric acid and then centrifuged at 2,000 x g using a continuous centrifuge (decanter) to obtain an insoluble fraction (acid precipitation card) and a soluble fraction (whey). It was. The acid precipitation curd was watered to a solid content of 10% by weight to obtain an acid precipitation curd slurry. This acid precipitation curd slurry was also used in the following examples, comparative examples, and experimental examples.
　固形分10重量%とした酸沈殿カードスラリーに、リン酸溶液でpH3.5に調整した後、50℃になるように加温した。この溶液に固形分あたり0.1重量%のAspergillus oryzae由来のプロテアーゼ（新日本化学社製「スミチームACP」1,000U/g）を加え、60分間加水分解を行った。反応後、85℃，20分間加熱し、酵素を失活させた。この加水分解物をpH3.5に調整し、固形分100gあたり８units相当のフィターゼ（ノボザイムズ社製「フィターゼノボ」）を加え、30分間作用させた。連続式直接加熱殺菌装置にて140℃，10秒間加熱した。これを噴霧乾燥し粉末状酸性可溶大豆たん白素材（A）（粗蛋白量91%）を得た。 The acid precipitation curd slurry having a solid content of 10% by weight was adjusted to pH 3.5 with a phosphoric acid solution and then heated to 50 ° C. To this solution, 0.1 wt% protease derived from Aspergillus oryzae (“Sumiteam ACP” manufactured by Shin Nippon Chemical Co., Ltd., 1,000 U / g) at a solid content was added and hydrolyzed for 60 minutes. After the reaction, the enzyme was inactivated by heating at 85 ° C. for 20 minutes. This hydrolyzate was adjusted to pH 3.5, phytase equivalent to 8 units per 100 g of solid content (“Phytase Novo” manufactured by Novozymes) was added and allowed to act for 30 minutes. Heated at 140 ° C for 10 seconds in a continuous direct heat sterilizer. This was spray-dried to obtain a powdery acidic soluble soybean protein material (A) (crude protein content 91%).
（実施例１）pH3.3でのプロテアーゼ分解・酸性可溶大豆たん白素材の調製
　比較例１の固形分10重量%の酸沈殿カードスラリーをリン酸でpH3.3に調整した後、50℃になるように加温した。この溶液に固形分あたり0.1重量%の「スミチームACP」を加え、60分間加水分解を行った。反応後、85℃，20分間加熱し、酵素を失活させた。この加水分解物をpH3.5に調整し、固形物あたり８unit相当の「フィターゼノボ」を加え、30分間酵素反応を行った。反応後、連続直接加熱殺菌装置にて140℃，10秒間加熱した。これを噴霧乾燥し粉末状酸性可溶大豆たん白素材（B）（粗蛋白量90%）を得た。 (Example 1) Preparation of protease-degrading / acid-soluble soybean protein material at pH 3.3 After adjusting the acid precipitation curd slurry of Comparative Example 1 having a solid content of 10% by weight to pH 3.3 with phosphoric acid, 50 ° C It was heated to become. To this solution, 0.1% by weight of “Sumiteam ACP” per solid content was added and hydrolyzed for 60 minutes. After the reaction, the enzyme was inactivated by heating at 85 ° C. for 20 minutes. This hydrolyzate was adjusted to pH 3.5, “phytase novo” corresponding to 8 units per solid was added, and the enzyme reaction was carried out for 30 minutes. After the reaction, it was heated at 140 ° C. for 10 seconds in a continuous direct heat sterilizer. This was spray-dried to obtain a powdery acidic soluble soybean protein material (B) (crude protein content 90%).
（実施例２）pH3.0でのプロテアーゼ分解・酸性可溶大豆たん白素材の調製
　比較例１の固形分10重量%の酸沈殿カードスラリーをリン酸でpH3.0に調整した後、50℃になるように加温した。この溶液に固形分あたり0.1重量%の「スミチームACP」を加え、60分間加水分解を行った。反応後、85℃，20分間加熱し、酵素を失活させた。この加水分解物に、固形物あたり８units相当の「フィターゼノボ」を加え、30分間酵素反応を行った。反応後、連続直接加熱殺菌装置にて140℃，10秒間加熱した。これを噴霧乾燥し粉末状酸性可溶大豆たん白素材（C）（粗蛋白量90%）を得た。 (Example 2) Preparation of protease-degrading / acid-soluble soy protein material at pH 3.0 After adjusting the acid precipitation curd slurry of Comparative Example 1 having a solid content of 10% by weight to pH 3.0 with phosphoric acid, 50 ° C It was heated to become. To this solution, 0.1% by weight of “Sumiteam ACP” per solid content was added and hydrolyzed for 60 minutes. After the reaction, the enzyme was inactivated by heating at 85 ° C. for 20 minutes. To this hydrolyzate, “phytase novo” corresponding to 8 units per solid was added, and an enzyme reaction was carried out for 30 minutes. After the reaction, it was heated at 140 ° C. for 10 seconds in a continuous direct heat sterilizer. This was spray-dried to obtain a powdery acidic soluble soybean protein material (C) (crude protein content 90%).
（比較例２）未分解・酸性可溶大豆たん白素材の調製
　比較例１の固形分10重量%の酸沈殿カードスラリーをリン酸溶液でpH3.5に調整した後、50℃で固形物あたり８unit相当の「フィターゼノボ」を加え、30分間作用させた。連続式直接加熱殺菌装置にて140℃，10秒加熱した。これらを噴霧乾燥し粉末状酸性可溶大豆たん白素材（D）（粗蛋白量92%）を得た。 (Comparative Example 2) Preparation of undegraded / acid-soluble soy protein material After adjusting the acid precipitation curd slurry having a solid content of 10% by weight of Comparative Example 1 to pH 3.5 with a phosphoric acid solution, “Phytase Novo” equivalent to 8 units was added and allowed to act for 30 minutes. Heated at 140 ° C for 10 seconds in a continuous direct heat sterilizer. These were spray-dried to obtain a powdery acidic soluble soybean protein material (D) (crude protein amount 92%).
（比較例３）pH3.5での植物プロテアーゼ分解・酸性可溶大豆たん白素材の調製
　比較例１の固形分10重量%の酸沈殿カードスラリーをリン酸でpH3.5に調整した後、50℃になるように加温した。この溶液に固形分あたり0.1重量%の「パパイン W-40」（天野エンザイム社製 400,000U/g）を加え、60分間加水分解を行った。反応後、85℃，20分間加熱し、酵素を失活させた。この加水分解物をpH3.5に調整し、固形物あたり８unit相当の「フィターゼノボ」を加え、pH3.5で30分間酵素反応を行った。反応後、連続直接加熱殺菌装置にて140℃，10秒間加熱した。これを噴霧乾燥し粉末状酸性可溶大豆たん白素材（E）（粗蛋白量91%）を得た。 (Comparative Example 3) Preparation of plant protease degradation / acid-soluble soybean protein material at pH 3.5 After adjusting the acid precipitation curd slurry having a solid content of 10% by weight of Comparative Example 1 to pH 3.5 with phosphoric acid, 50 Heated to 0 ° C. To this solution, 0.1% by weight of “papain W-40” (400,000 U / g, manufactured by Amano Enzyme) per solid content was added, and hydrolysis was performed for 60 minutes. After the reaction, the enzyme was inactivated by heating at 85 ° C. for 20 minutes. The hydrolyzate was adjusted to pH 3.5, “phytase novo” corresponding to 8 units per solid was added, and the enzyme reaction was carried out at pH 3.5 for 30 minutes. After the reaction, it was heated at 140 ° C. for 10 seconds in a continuous direct heat sterilizer. This was spray-dried to obtain a powdery acidic soluble soybean protein material (E) (crude protein content 91%).
（比較例４）中性プロテアーゼ分解・酸性可溶大豆たん白素材の調製
　比較例１の固形分10重量%の酸沈殿カードスラリーを希水酸化ナトリウム溶液でpH6.5に調整した後、連続式直接加熱殺菌装置にて140℃，10秒加熱した。この溶液を50℃として固形分あたり0.1重量%のBacillus属由来のプロテアーゼ（ノボザイム社製「プロタメックス」1.5AU（アンソン単位）/g）を加え、60分間加水分解を行った。反応後、85℃，20分間加熱し、酵素を失活させた。この加水分解物にリン酸を添加しpH3.5に調整した後、固形物あたり８unit相当の「フィターゼノボ」を加え、30分間作用させた。連続式直接加熱殺菌装置にて140℃，10秒加熱した。これらを噴霧乾燥し粉末状酸性可溶大豆たん白素材（F）（粗蛋白量80%）を得た。 Comparative Example 4 Preparation of Neutral Protease Decomposition / Acid Soluble Soy Protein Material After adjusting the acid precipitation curd slurry having a solid content of 10% by weight of Comparative Example 1 to pH 6.5 with dilute sodium hydroxide solution, continuous Heated at 140 ° C for 10 seconds in a direct heat sterilizer. This solution was adjusted to 50 ° C., and 0.1 wt% of protease derived from the genus Bacillus (“Protamex” 1.5 AU (Anson unit) / g, manufactured by Novozyme) per solid content was added, followed by hydrolysis for 60 minutes. After the reaction, the enzyme was inactivated by heating at 85 ° C. for 20 minutes. After adding phosphoric acid to this hydrolyzate and adjusting to pH 3.5, "phytase Novo" corresponding to 8 units per solid was added and allowed to act for 30 minutes. Heated at 140 ° C for 10 seconds in a continuous direct heat sterilizer. These were spray-dried to obtain a powdery acidic soluble soybean protein material (F) (crude protein amount 80%).
（比較例５）分解度の高い酸性可溶大豆たん白素材の調製
　比較例１の固形分10重量%の酸沈殿カードスラリーをリン酸溶液でpH3.0に調整した後、50℃になるように加温した。この溶液に固形分あたり0.5重量%の「スミチームACP」を加え、60分間加水分解を行った。反応後、85℃，20分間加熱し、酵素を失活させた。この加水分解物に、固形分100gあたり８units相当の「フィターゼノボ」を加え、30分間作用させた。連続式直接加熱殺菌装置にて140℃，10秒間加熱した。これを噴霧乾燥し粉末状酸性可溶大豆たん白素材（G）（粗蛋白量90%）を得た。 (Comparative Example 5) Preparation of acid-soluble soybean protein material having a high degree of decomposition After adjusting the acid precipitation curd slurry having a solid content of 10% by weight of Comparative Example 1 to pH 3.0 with a phosphoric acid solution, the temperature becomes 50 ° C. Warmed to. To this solution, 0.5% by weight of “Sumiteam ACP” per solid content was added and hydrolyzed for 60 minutes. After the reaction, the enzyme was inactivated by heating at 85 ° C. for 20 minutes. To this hydrolyzate, “phytase novo” corresponding to 8 units per 100 g of solid content was added and allowed to act for 30 minutes. Heated at 140 ° C for 10 seconds in a continuous direct heat sterilizer. This was spray-dried to obtain a powdery acidic soluble soybean protein material (G) (crude protein content 90%).
（実施例３）pH3.0でのプロテアーゼ分解・酸性可溶大豆たん白素材の調製２
　実施例２と同様に調製し、粉末状酸性可溶大豆たん白素材（H）（粗蛋白量90%）を得た。ただし、「スミチームACP」を溶液に固形分あたり0.25重量%とし、60分間加水分解を行った。 (Example 3) Preparation of protease-degrading / acid-soluble soybean protein material at pH 3.0 2
Preparation was carried out in the same manner as in Example 2 to obtain a powdery acidic soluble soybean protein material (H) (crude protein content 90%). However, “Sumiteam ACP” was added to the solution at a concentration of 0.25 wt% per solid, and hydrolysis was performed for 60 minutes.
（官能評価および品質の比較）
　上記、酸性可溶大豆たん白素材（A）～（H）について官能評価、品質測定を実施した（表１）。官能評価は、５重量%水溶液を調製し、「渋味」「苦味」を評価した。それぞれの項目について、感じない「１点」、ほとんど感じない「２点」、感じるが不快ではない「３点」、感じる（不快である）「４点」、強く感じる「５点」に基づき、20人のパネラーによって官能評価を行い、その平均値を示した。 (Sensory evaluation and quality comparison)
Sensory evaluation and quality measurement were carried out on the acid-soluble soybean protein materials (A) to (H) (Table 1). Sensory evaluation prepared 5 weight% aqueous solution, and evaluated "astringency" and "bitterness". For each item, based on "1 point" that you do not feel, "2 points" that you hardly feel, "3 points" that you feel but are uncomfortable, "4 points" that you feel (discomfort), "5 points" that you feel strongly, Sensory evaluation was performed by 20 panelists, and the average value was shown.
　pH3.4以下での分解を開始した（B）（C）では、渋味，苦味とも低減しており酸性飲食物に用いるに好適であった。未分解の（D）は渋味が非常に強く感じられた。pH3.4を超えたpHで分解を開始した（A）（E）は、（D）に対して渋味が低下したが不快ではないレベルまで低下させることはできず、苦味が不快に感じられるレベルで発生していた。中性分解の（F）は（E）より渋味が低下するものの、やや問題があり、大豆臭が感じられた。よって、(A）（D）（E）（F）は酸性飲食物に好ましく使用できるレベルには達しなかった。酵素量を増やし分解程度を上げた（G）および（H）について、（G）ではTCA可溶化率が45%を超えており、苦味が不快に感じられるレベルまで上昇しており、本発明の目的に合致しないものだったが、TCA可溶化率が45%以下である（H）は、良好な風味を有していた。 In (B) and (C) where decomposition was started at pH 3.4 or lower, both astringency and bitterness were reduced, which was suitable for use in acidic foods and drinks. Undegraded (D) felt a very strong astringency. (A) (E), which started degrading at a pH exceeding 3.4, has a bitter taste compared to (D) but cannot be reduced to a level that is not unpleasant, and bitterness is felt uncomfortable. Had occurred at the level. Neutral degradation (F) was slightly less astringent than (E), but had some problems and felt a soy odor. Therefore, (A), (D), (E), and (F) did not reach the level that can be preferably used for acidic foods and drinks. For (G) and (H), which increased the amount of enzyme and increased the degree of degradation, in (G), the TCA solubilization rate exceeded 45% and increased to a level at which bitterness was felt uncomfortable. Although it did not meet the purpose, a TCA solubilization rate of 45% or less (H) had a good flavor.
（表１）官能評価および各品質の比較
Figure JPOXMLDOC01-appb-I000001
(Table 1) Sensory evaluation and comparison of each quality
Figure JPOXMLDOC01-appb-I000001
（実験例１）ゲル濾過による分子量分布
　また、（B）～（F）について、ゲル濾過による比較を行った。その結果を表１および図１に示した。渋みが認められない（B）（C）（H）で、質量数150kda以上の画分の含量がいずれも18%以下と低い値だったことより、高い質量数の画分と渋みは関係を有することが判った。尚、ゲル濾過測定は前述した系を用い分子量マーカーであるγ-globlin（MP Biomedicals, Inc.製 分子量150kDa）の溶出時間8.1830分より早い溶出時間のピーク面積の割合を測定し、質量数150kDa以上の画分の含量とした。試料は蛋白質として0.1重量%になる様に溶離液中に溶解させた後、セルロースアセテートタイプメンブレンフィルター（DISMIC-13CP 0.20μm , Advantec社製）を通し沈殿を除去した後、20μlをインジェクションした。 (Experimental example 1) Molecular weight distribution by gel filtration Further, (B) to (F) were compared by gel filtration. The results are shown in Table 1 and FIG. No astringency was observed (B), (C), and (H), and the fractions with a mass number of 150 kda or more were all low values of 18% or less. It was found to have. For gel filtration measurement, the ratio of the peak area of elution time earlier than 8.1830 minutes was measured for the molecular weight marker γ-globlin (MP Biomedicals, Inc. molecular weight 150 kDa) using the system described above, and the mass number was 150 kDa or more. It was set as the content of the fraction. The sample was dissolved in the eluent so as to be 0.1% by weight as protein, and after passing through a cellulose acetate type membrane filter (DISMIC-13CP 0.20 μm, manufactured by Advantec) to remove the precipitate, 20 μl was injected.
　尚、HPLC本体は日本分光社製の機器を用い、送液ポンプとして「PU-2089 Plus」、検出器として「UV-2077 Plus」、カラムオーブンとして「CO‐2,065 Plus」、システムコントローラーとして「LC-Net II/ADC」、分析ソフトとして「ChromNAV」を用いた。また、クロマトグラムの波形処理は、ChromNAVでスロープ感度10.00μV/sec，スロープ幅0.1min，最小面積10000μV・sec，最小高さ1000μV，ドリフト0.000μV/min，スロープ幅倍化時間0.0000minとした。 The HPLC main unit uses equipment manufactured by JASCO Corporation, “PU-2089 Plus” as the liquid feed pump, “UV-2077 Plus” as the detector, “CO-2,065 Plus” as the column oven, and “LC” as the system controller. -Net II / ADC "and" ChromNAV "as analysis software. The chromatogram waveform processing was ChromNAV with a slope sensitivity of 10.00 μV / sec, a slope width of 0.1 min, a minimum area of 10,000 μV · sec, a minimum height of 1000 μV, a drift of 0.000 μV / min, and a slope width doubling time of 0.0000 min.
（実験例２）SDS電気泳動による比較
　（C）～（F）について、SDS-ポリアクリルアミド電気泳動に供し、蛋白質の挙動を調べた。電気泳動は、Laemmliの方法［Nature，227巻，680-685（1970）］に準じた。すなわち、試料の５重量%水溶液10μLと、トリス-SDS-β-MEサンプル処理液（コスモバイオ社製）90μLを混合後100℃で5分間加熱し、SDS-トリス-グリシン泳動バッファー（コスモバイオ社製）中10～20%グラジエントゲルを使用し、25mAの定流で１時間実施した。泳動後、ゲルをページブルー83（CBB R-250）染色液（コスモバイオ社製）にて染色した。 (Experimental Example 2) Comparison by SDS Electrophoresis (C) to (F) were subjected to SDS-polyacrylamide electrophoresis to examine the behavior of proteins. Electrophoresis was in accordance with the Laemmli method [Nature, Vol. 227, 680-685 (1970)]. That is, 10 μL of a 5% by weight aqueous solution of a sample and 90 μL of Tris-SDS-β-ME sample treatment solution (manufactured by Cosmo Bio) were mixed and heated at 100 ° C. for 5 minutes, and SDS-Tris-Glycine running buffer (Cosmo Bio 10-20% gradient gel was used, and a constant flow of 25 mA was used for 1 hour. After electrophoresis, the gel was stained with Page Blue 83 (CBB R-250) staining solution (manufactured by Cosmo Bio).
　図２に示すように、渋味が強く感じられた、未分解の（D）およびpH3.5で分解を開始した（E）では、pH3.0で分解を開始した（C）と比較して7Sグロブリン，11Sグロブリン等の成分が分解されずバンドが認められた。また、ゲル濾過による150kda以上の画分の含量も高いことから（表１）、（D）（E）では、これら未分解の7S，11Sの蛋白質が渋みに関与していると想定されるとともに、（E）のpH3.5で分解を開始した条件では、TCA可溶化率は高いにも関わらず、7Sグロブリン，11Sグロブリンについて、適切な分解が行われないことが判った。一方で中性分解の（F）は（C）に比べ電気泳動で認められるバンドが更に薄く、分解度が高いにもかかわらず、ゲル濾過による150kda以上の画分の含量が21.8%と高く（表１，図１）、（C）に比べて渋味がまだ感じられ、更に粗蛋白量も他より低かった。以上の検討から、苦みの出ない分解条件の中で、単純に加水分解を行うだけでは、SS結合等による高分子化物を解離させることはできず、渋味低下につながらないことが確認された。 As shown in FIG. 2, the undegraded (D) and the decomposition started at pH 3.5 (E), where astringency was felt strongly, compared to the decomposition started at pH 3.0 (C). Components such as 7S globulin and 11S globulin were not degraded and bands were observed. In addition, since the content of fractions of 150 kda or more by gel filtration is high (Table 1), (D) and (E), it is assumed that these undegraded 7S and 11S proteins are involved in astringency. (E) Under the conditions where the decomposition was started at pH 3.5, it was found that the 7S globulin and 11S globulin were not properly decomposed even though the TCA solubilization rate was high. On the other hand, (F) of neutral degradation has a thinner band observed by electrophoresis than (C), and even though it has a high degree of degradation, the content of fractions of 150 kda or higher by gel filtration is as high as 21.8% ( Compared with Table 1, FIG. 1), (C), astringency was still felt, and also the amount of crude protein was lower than others. From the above examination, it was confirmed that the polymerized product due to SS bond or the like cannot be dissociated by simply performing hydrolysis under the decomposition conditions that do not cause bitterness, and it does not lead to a decrease in astringency.
（実施例４）11Sグロブリンを原料とした、pH3.0での分解による酸性可溶大豆たん白素材の調製
　分離大豆たん白より11Sグロブリンを、国際公開WO2006/129647号公報に記載の製造方法を用いて分離し、11Sグロブリン濃縮素材を得た。すなわち、低変性脱脂大豆1kgに70重量%含水アルコールを100g噴霧しながら混合し、脱脂大豆の品温が70℃になるように密閉容器の外側を加熱し、30分維持した。得られた加工脱脂大豆１重量部に水７重量部を加え、希水酸化ナトリウム溶液でpH7.5に調整しつつ室温で30分撹拌しながら抽出後、1,000×gで遠心分離しオカラおよび不溶分を分離し、脱脂豆乳Ａを得た。オカラおよび不溶物に水５重量部を加え、室温で30分撹拌しながら抽出後、1,000×gで遠心分離しオカラおよび不溶分を分離し、脱脂豆乳Ｂを得た。脱脂豆乳ＡとBを混合し、塩酸を用いてpH5.8に調整し、1,000×gで遠心分離し、不溶性画分である11Sグロブリン濃縮素材を得た。この11Sグロブリン濃縮素材に固形分10重量%になるように加水しpH3.0に調整し、固形分あたり0.1重量%の「スミチームACP」を添加し50℃で60分間加水分解を行った。反応後、85℃，20分間加熱し、酵素を失活させた。この加水分解物をpH3.5に調整し、固形分100gあたり８units相当の「フィターゼノボ」を加え、30分間作用させた。連続式直接加熱殺菌装置にて140℃，10秒間加熱した。これを噴霧乾燥し、粉末状酸性可溶大豆たん白素材（I）（粗蛋白量90%）を得た。 (Example 4) Preparation of acidic soluble soybean protein material by decomposition at pH 3.0 using 11S globulin as a raw material 11S globulin from isolated soybean protein was prepared by the production method described in International Publication WO2006 / 129647. To obtain 11S globulin-enriched material. That is, 1 kg of low-denatured defatted soybean was mixed while spraying 100 g of 70 wt% hydrous alcohol, and the outside of the sealed container was heated so that the product temperature of the defatted soybean was 70 ° C. and maintained for 30 minutes. 7 parts by weight of water was added to 1 part by weight of the processed defatted soybeans, extracted with stirring at room temperature for 30 minutes while adjusting to pH 7.5 with dilute sodium hydroxide solution, centrifuged at 1,000 xg, and okara and insoluble. Minutes were separated to obtain defatted soymilk A. 5 parts by weight of water was added to okara and insoluble matter, extracted while stirring at room temperature for 30 minutes, and centrifuged at 1,000 × g to separate okara and insoluble matter to obtain defatted soymilk B. Skim soymilk A and B were mixed, adjusted to pH 5.8 using hydrochloric acid, and centrifuged at 1,000 × g to obtain an 11S globulin-enriched material that was an insoluble fraction. The 11S globulin concentrated material was added with water to a solid content of 10% by weight, adjusted to pH 3.0, 0.1% by weight of “Sumiteam ACP” was added to the solid content, and hydrolysis was performed at 50 ° C. for 60 minutes. After the reaction, the enzyme was inactivated by heating at 85 ° C. for 20 minutes. This hydrolyzate was adjusted to pH 3.5, “phytase Novo” corresponding to 8 units per 100 g of solid content was added, and allowed to act for 30 minutes. Heated at 140 ° C for 10 seconds in a continuous direct heat sterilizer. This was spray-dried to obtain powdered acidic soluble soybean protein material (I) (crude protein content 90%).
（比較例６）11Sグロブリンを原料とした、未分解の酸性可溶大豆たん白素材の調製
　実施例４に記載の固形分10重量%の11Sグロブリン濃縮素材をpH3.0に調整し、140℃，10秒間加熱後噴霧乾燥し、酸性可溶化処理を行った11Sグロブリン未分解物（J）（粗蛋白量92%）を得た。 (Comparative Example 6) Preparation of undegraded acidic soluble soybean protein material using 11S globulin as a raw material The 11S globulin concentrated material having a solid content of 10% by weight described in Example 4 was adjusted to pH 3.0, and 140 ° C. , Heated for 10 seconds and spray-dried to obtain an 11S globulin undegraded product (J) (crude protein amount 92%) subjected to acid solubilization treatment.
（比較例７）11Sグロブリンを原料とした、中性分解による酸性可溶大豆たん白素材の調製
　実施例４に記載の固形分10重量%の11Sグロブリン濃縮素材をpH7.0に調整し、この溶液を50℃として固形分あたり0.1%の「プロタメックス」を加え、60分間加水分解を行った。反応後、85℃，20分間加熱し、酵素を失活させた。この加水分解物にリン酸を添加しpH3.5に調整した後、固形物あたり８unit相当の「フィターゼノボ」を加え、30分間作用させた。連続式直接加熱殺菌装置にて140℃，10秒加熱後噴霧乾燥し、酸性可溶大豆たん白素材（K）（粗蛋白量80%）を得た。 (Comparative Example 7) Preparation of acid-soluble soybean protein material by neutral decomposition using 11S globulin as a raw material The 11S globulin concentrated material having a solid content of 10% by weight described in Example 4 was adjusted to pH 7.0. The solution was adjusted to 50 ° C. and 0.1% “Protamex” per solid content was added, followed by hydrolysis for 60 minutes. After the reaction, the enzyme was inactivated by heating at 85 ° C. for 20 minutes. After adding phosphoric acid to this hydrolyzate and adjusting to pH 3.5, "phytase Novo" corresponding to 8 units per solid was added and allowed to act for 30 minutes. The mixture was heated at 140 ° C. for 10 seconds in a continuous direct heat sterilizer and then spray-dried to obtain an acidic soluble soybean protein material (K) (crude protein content 80%).
（実施例５）7Sグロブリンを原料とした、pH3.0での分解による酸性可溶大豆たん白素材の調製
　分離大豆たん白より7Sグロブリンを国際公開WO2002/28198号公報に記載の製造方法を用いて分離し、7Sグロブリン濃縮素材を得た。すなわち、低変性脱脂大豆１重量部に水10重量部を加え、希水酸化ナトリウム溶液でpH7.0に調整しつつ室温で１時間撹拌しながら抽出後、1,000×gで遠心分離しオカラおよび不溶分を分離し、脱脂豆乳を得た。この脱脂豆乳を塩酸を用いてpH4.2に調整し、40℃に加温後冷却し、pH5.8に調整後遠心分離して可溶性画分を得た。可溶性画分を塩酸を用いてpH4.9に調整し、1,000×gで遠心分離して沈殿である7Sグロブリン濃縮素材を得た。7Sグロブリン濃縮素材に固形分10重量%になるように加水しpH3.0に調整し、固形分あたり0.1%の「スミチームACP」を添加し50℃で60分間加水分解を行った。反応後、85℃，20分間加熱し、酵素を失活させた。この加水分解物に固形分100gあたり８units相当の「フィターゼノボ」を加え、30分間作用させた。連続式直接加熱殺菌装置にて140℃，10秒間加熱した。これを噴霧乾燥し粉末状酸性可溶大豆たん白素材（L）（粗蛋白量90%）を得た。 (Example 5) Preparation of acidic soluble soy protein material by decomposition at pH 3.0 using 7S globulin as a raw material 7S globulin from separated soy protein was produced using the production method described in International Publication WO2002 / 28198 And 7S globulin-enriched material was obtained. That is, 10 parts by weight of water is added to 1 part by weight of low-denatured defatted soybeans, extracted with stirring for 1 hour at room temperature while adjusting to pH 7.0 with dilute sodium hydroxide solution, and centrifuged at 1,000 xg for okara and insoluble. Minutes were separated to obtain defatted soymilk. The defatted soymilk was adjusted to pH 4.2 using hydrochloric acid, heated to 40 ° C., cooled, adjusted to pH 5.8 and centrifuged to obtain a soluble fraction. The soluble fraction was adjusted to pH 4.9 with hydrochloric acid and centrifuged at 1,000 × g to obtain a 7S globulin-enriched material that was a precipitate. The 7S globulin concentrate was hydrated to a solid content of 10% by weight, adjusted to pH 3.0, 0.1% “Sumiteam ACP” was added per solid content, and hydrolysis was performed at 50 ° C. for 60 minutes. After the reaction, the enzyme was inactivated by heating at 85 ° C. for 20 minutes. “Phytase Novo” corresponding to 8 units per 100 g of solid content was added to this hydrolyzate and allowed to act for 30 minutes. Heated at 140 ° C for 10 seconds in a continuous direct heat sterilizer. This was spray-dried to obtain powdered acidic soluble soybean protein material (L) (crude protein content 90%).
（官能評価および品質の比較）
　上記、酸性可溶大豆たん白素材（C）および（I）～（L）について、官能評価、品質測定を実施した（表２）。尚、11Sグロブリンに富む素材についてpH3.4以下で分解を開始した（I）は、150kda以上の画分の含量が15.5%とやや高いが、（C）に比べ若干渋味が強いものの、不快に感じる程ではなかった。また、苦味が低減しており、酸性飲食物に使用するに好適であった。未分解の（J）は渋味が強く感じられた。中性分解の（K）は（J）に比べ渋味が低減したが不快に感じられるレベルであった。また、7Sグロブリンに富む素材から調製した（L）は渋味，苦味が低く、酸性飲食物に使用できるレベルとなった。以上から、11Sグロブリンおよび7Sグロブリンに富む素材を原料とする場合でも実施例２同様、pH3.4以下で酵素分解を開始することにより、酸性飲食物に使用できるレベルまで、酸性可溶大豆たん白素材の渋味を低下させることができた。 (Sensory evaluation and quality comparison)
Sensory evaluation and quality measurement were performed on the above-mentioned acidic soluble soybean protein materials (C) and (I) to (L) (Table 2). The 11S globulin-rich material started to decompose at pH 3.4 or lower (I), which has a slightly higher content of 15.5% of the fraction of 150 kda or higher, but slightly astringent compared to (C), but uncomfortable. It wasn't enough to feel. Moreover, the bitterness was reduced and it was suitable for using for acidic food and drink. Undecomposed (J) felt strong astringency. Neutral degradation (K) was less astringent than (J), but felt uncomfortable. In addition, (L) prepared from a material rich in 7S globulin has low astringency and bitterness, and can be used for acidic foods. From the above, even when using a material rich in 11S globulin and 7S globulin as a raw material, as in Example 2, by starting enzymatic degradation at pH 3.4 or lower, acidic soluble soybean protein to a level that can be used for acidic foods and drinks. The astringency of the material could be reduced.
（表２）官能評価および各品質の比較
Figure JPOXMLDOC01-appb-I000002
(Table 2) Sensory evaluation and comparison of each quality
Figure JPOXMLDOC01-appb-I000002
（実施例６）酸性粉末飲料の調製
　実施例２で調製した粉末状酸性可溶大豆たん白素材（C）を66.7部、グラニュー糖を18.6部，グレープフルーツ粉末果汁（焼津水産化学社製）12.4部，グレープフルーツ香料（高砂香料社製）2.1部，甘味料（スクラロース・三栄源エフエフアイ社製）0.2部をよく混合させ、蛋白質含有酸性粉末飲料を得た。この粉末15gを水185gに加え、シェーカーにてよく混合させ飲料とした。本飲料を官能にて評価したが、適当な酸味を有し、従来の酸性可溶大豆たん白素材特有の渋味を感じず、かつ苦味も感じず、大豆臭も感じず、喉越しも良好で嗜好性に優れるものであった。 (Example 6) Preparation of acidic powder beverage 66.7 parts of powdered acidic soluble soybean protein material (C) prepared in Example 2, 18.6 parts of granulated sugar, 12.4 parts of grapefruit powder juice (made by Yaizu Suisan Chemical Co., Ltd.) , Grapefruit fragrance (manufactured by Takasago Fragrance Co., Ltd.) 2.1 parts and sweetener (Sucralose, Saneigen FFI Co., Ltd.) 0.2 part were mixed well to obtain a protein-containing acidic powder drink. 15 g of this powder was added to 185 g of water and mixed well with a shaker to obtain a beverage. Although this beverage was evaluated by sensory evaluation, it has an appropriate acidity, does not feel the astringent taste peculiar to conventional acidic soluble soybean protein materials, does not feel bitterness, does not feel soybean odor, and feels good over the throat It was excellent in palatability.
（実施例７）酸性ゼリー状食品の調製
　実施例２で調製した粉末状酸性可溶大豆たん白素材（C）を5.0重量部，果糖ブドウ糖液（日本コーンスターチ社製）8.0部，５倍濃縮パイナップル果汁（フードマテリアル社製）2.5部，寒天（伊那寒天社製）0.3部，パイナップル香料（高砂香料社製）0.2部，水84.0部の配合で、寒天以外を十分に撹拌混合しクエン酸ナトリウムでpH3.6に調整後、95℃達温加熱殺菌を行い、加熱により膨潤させた寒天液と高温で混ぜ合わせ、チアーパックに充填後一晩冷蔵によりゲル化させゼリー飲料を調製した。得られたゼリー飲料は、渋みを感じず、食感も良好であった。 (Example 7) Preparation of acidic jelly-like food 5.0 parts by weight of powdered acid-soluble soybean protein material (C) prepared in Example 2, 8.0 parts of fructose glucose liquid (manufactured by Cornstarch Japan), 5-fold concentrated pineapple Mixing fruit juice (Food Material Co., Ltd.) 2.5 parts, agar (Ina Agar Co., Ltd.) 0.3 parts, pineapple fragrance (Takasago Fragrance Co., Ltd.) 0.2 parts, water 84.0 parts. The mixture was adjusted to pH 3.6, sterilized by heating at 95 ° C., mixed with agar liquid swollen by heating at high temperature, filled in a cheer pack, and gelled by refrigeration overnight to prepare a jelly beverage. The obtained jelly beverage did not feel astringent and had a good texture.
（実施例８）酸性嚥下食の調製
　実施例２で調製した粉末状酸性可溶大豆たん白素材（C）4.3重量部，グラニュー糖12.5重量部，醗酵乳酸ドロマイト（明治乳業製）1.25重量部，ビタミン強化剤0.125重量部，酒石酸0.1重量部，ミネラル酵母製剤「ミックスミネラルイーストＣ」0.1重量部、と「ミックスミネラルイーストＭ」0.025重量部（エル・エスコーポレーション製），ステビア製剤を0.06重量部を粉体混合し、水46.04重量部に分散溶解させた。溶解には高速乳化分散機を用いた。次いで、硬化菜種油4.3重量部を少しずつ加え、油分が分離しないように攪拌した（溶液１）。
　寒天製剤「アガーミックス＃24」（青葉化成製）1.3重量部を水25重量部に分散させ、湯煎で10分間加熱し溶解させた（溶液２）。溶液１を攪拌しながら溶液２を混合した。そこに透明濃縮マンゴー果汁4.0重量部、色素「βカロテン10C」（三菱化学フーズ製）0.1重量部、香料0.9重量部を加えて混合し、スタンディングパウチに充填した。これを85℃で30分間加熱殺菌して、流水にて冷却しゼリー状の嚥下食（pH3.90） を得た。
　このものは、三大栄養素であるたん白，脂質，糖質を含み、１gで1.2kcalを摂取でき、またその他の体に必要な栄養素、すなわちカルシウム，マグネシウム，微量元素ミネラル，ビタミン，食物繊維を全て含み、やわらかく滑らかな食感で、しかも咽頭部でのべたつき、及び離水の少ないソフトゼリーで、嚥下困難な高齢者や病態者が誤嚥することなく摂食することができるものであった。 (Example 8) Preparation of acidic swallow food Powdered acidic soluble soybean protein material prepared in Example 2 (C) 4.3 parts by weight, granulated sugar 12.5 parts by weight, fermented lactate dolomite (Meiji Dairies) 1.25 parts by weight, Vitamin fortifier 0.125 parts by weight, tartaric acid 0.1 parts by weight, mineral yeast preparation “Mix Mineral Yeast C” 0.1 part by weight, “Mix Mineral Yeast M” 0.025 part by weight (manufactured by LS Corporation), stevia preparation 0.06 part by weight The powder was mixed and dispersed and dissolved in 46.04 parts by weight of water. A high-speed emulsification disperser was used for dissolution. Next, 4.3 parts by weight of hardened rapeseed oil was added little by little and stirred so that the oil was not separated (solution 1).
1.3 parts by weight of an agar preparation “Agar Mix # 24” (manufactured by Aoba Kasei) was dispersed in 25 parts by weight of water and dissolved by heating in a hot water bath for 10 minutes (solution 2). Solution 2 was mixed while solution 1 was stirred. Thereto, 4.0 parts by weight of transparent concentrated mango juice, 0.1 part by weight of the dye “β-carotene 10C” (manufactured by Mitsubishi Chemical Foods) and 0.9 part by weight of fragrance were added and mixed, and filled into a standing pouch. This was sterilized by heating at 85 ° C. for 30 minutes and cooled with running water to obtain a jelly-like swallowed food (pH 3.90).
It contains protein, lipid, and sugar, which are the three major nutrients, and can consume 1.2kcal in 1g, and other nutrients necessary for the body: calcium, magnesium, trace element minerals, vitamins, and dietary fiber. Including soft jelly with a soft and smooth texture, and a soft jelly with little stickiness in the pharynx, it was able to be eaten by an elderly person or a patient who had difficulty in swallowing without aspiration.
（実施例９）酸性ホイップクリームの調製
　実施例２で調製した粉末状酸性可溶大豆たん白素材（C）を2.0重量部、蔗糖脂肪酸エステルS-570（三菱化学フーズ社製）0.2重量部に、水52.6重量部を加え溶解させ、更にpHを3.5として水相を調製した。別途、パーム油中融点画分23重量部、硬化ヤシ油22重量部、大豆レシチン（辻製油社製）0.2重量部を混合し、油相を調製した。水相と油相を混合し、70℃，15分間ホモミキサーで撹拌し予備乳化した後、1MPaの圧力で均質化処理後、144℃，４秒間、蒸気直接加熱による殺菌処理を行った。続けて4MPaの圧力で均質化処理し、直ちに冷却し５℃，24時間保持することで、酸性クリームを調製した。このクリームをホイップしたところ、３分間のホイップタイムで90%と高いオーバーランを示し、組織も滑らかで、併せて爽やかな酸味を示し、良好であった。 (Example 9) Preparation of acidic whipped cream To 2.0 parts by weight of powdered acidic soluble soybean protein material (C) prepared in Example 2, 0.2 parts by weight of sucrose fatty acid ester S-570 (manufactured by Mitsubishi Chemical Foods) Then, 52.6 parts by weight of water was added and dissolved, and an aqueous phase was prepared at a pH of 3.5. Separately, 23 parts by weight of a middle melting point fraction of palm oil, 22 parts by weight of hardened coconut oil, and 0.2 parts by weight of soybean lecithin (manufactured by Sakai Oil Co., Ltd.) were mixed to prepare an oil phase. The aqueous phase and the oil phase were mixed, stirred with a homomixer at 70 ° C. for 15 minutes and pre-emulsified, then homogenized at a pressure of 1 MPa, and then sterilized by direct steam heating at 144 ° C. for 4 seconds. Subsequently, the mixture was homogenized at a pressure of 4 MPa, immediately cooled, and kept at 5 ° C. for 24 hours to prepare an acidic cream. When this cream was whipped, it showed a high overrun of 90% at a whipping time of 3 minutes, the structure was smooth, and a refreshing sour taste was also obtained.
　本発明により、渋味も苦味も少なく風味に優れ、かつ酸性飲食物用として求められる低粘度，高溶解性，高安定性などの物理化学的特性を満たす、たん白素材を製造することができる。この酸性可溶大豆たん白素材を用いることにより、従来より風味に優れた蛋白質含有酸性飲食物を製造することができる。 According to the present invention, it is possible to produce a protein material that has little astringency and bitterness, excellent flavor, and satisfies the physicochemical properties such as low viscosity, high solubility, and high stability required for acidic foods and drinks. . By using this acidic soluble soybean protein material, protein-containing acidic foods and drinks that have a better flavor than before can be produced.

Claims (4)

  1. １重量%SDS含有緩衝液（pH7.0）を用いたゲル濾過において、220nmで検出される、質量数150kDa以上の画分の含量が18%以下で且つTCA（0.22M）可溶化率が45%以下の、酸性可溶大豆たん白素材。 In gel filtration using a buffer solution (pH 7.0) containing 1% by weight SDS, the content of a fraction having a mass number of 150 kDa or more detected at 220 nm is 18% or less, and the TCA (0.22M) solubilization rate is 45. Less than% acidic soluble soy protein material.
  2. pH3.4以下でプロテアーゼ処理を開始し、TCA（0.22M）可溶化率を10%以上，45%以下とすることを特徴とする、酸性可溶大豆たん白素材の製造方法。 A method for producing an acidic soluble soybean protein material, characterized in that protease treatment is started at pH 3.4 or lower, and TCA (0.22M) solubilization rate is 10% or more and 45% or less.
  3. 酸性可溶大豆たん白素材の製造が、フィターゼ処理および100℃を超える酸性加熱処理によるものである、請求項２に記載の酸性可溶大豆たん白素材の製造方法。 The method for producing an acid-soluble soybean protein material according to claim 2, wherein the production of the acid-soluble soybean protein material is performed by a phytase treatment and an acidic heat treatment exceeding 100 ° C.
  4. 請求項１に記載の酸性可溶大豆たん白素材を用いた、酸性飲食物。 Acidic food and drink using the acid-soluble soybean protein material according to claim 1.
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