CN116997264A - Method for preparing processed soybean beverage - Google Patents
Method for preparing processed soybean beverage Download PDFInfo
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- CN116997264A CN116997264A CN202280017468.4A CN202280017468A CN116997264A CN 116997264 A CN116997264 A CN 116997264A CN 202280017468 A CN202280017468 A CN 202280017468A CN 116997264 A CN116997264 A CN 116997264A
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- soybean
- aspergillus
- protease
- beverage
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- 238000000034 method Methods 0.000 title claims description 25
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- 241001465318 Aspergillus terreus Species 0.000 description 2
- 241000203233 Aspergillus versicolor Species 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- SUZLHDUTVMZSEV-UHFFFAOYSA-N Deoxycoleonol Natural products C12C(=O)CC(C)(C=C)OC2(C)C(OC(=O)C)C(O)C2C1(C)C(O)CCC2(C)C SUZLHDUTVMZSEV-UHFFFAOYSA-N 0.000 description 2
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- 101710180012 Protease 7 Proteins 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- 241000222354 Trametes Species 0.000 description 2
- 229940091771 aspergillus fumigatus Drugs 0.000 description 2
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- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 description 2
- 239000012295 chemical reaction liquid Substances 0.000 description 2
- OHCQJHSOBUTRHG-UHFFFAOYSA-N colforsin Natural products OC12C(=O)CC(C)(C=C)OC1(C)C(OC(=O)C)C(O)C1C2(C)C(O)CCC1(C)C OHCQJHSOBUTRHG-UHFFFAOYSA-N 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
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- 239000000706 filtrate Substances 0.000 description 2
- 235000021552 granulated sugar Nutrition 0.000 description 2
- 230000002779 inactivation Effects 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- DNIAPMSPPWPWGF-UHFFFAOYSA-N monopropylene glycol Natural products CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 2
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- IITIZHOBOIBGBW-UHFFFAOYSA-N 3-ethyl-2h-1,3-benzothiazole Chemical compound C1=CC=C2N(CC)CSC2=C1 IITIZHOBOIBGBW-UHFFFAOYSA-N 0.000 description 1
- 241000349731 Afzelia bipindensis Species 0.000 description 1
- 102000004400 Aminopeptidases Human genes 0.000 description 1
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- 241000134821 Aspergillus caesiellus Species 0.000 description 1
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- 101001091385 Homo sapiens Kallikrein-6 Proteins 0.000 description 1
- 208000033830 Hot Flashes Diseases 0.000 description 1
- 206010060800 Hot flush Diseases 0.000 description 1
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- 241001465754 Metazoa Species 0.000 description 1
- 241000235395 Mucor Species 0.000 description 1
- 241000221960 Neurospora Species 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
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- 229920001214 Polysorbate 60 Polymers 0.000 description 1
- 241000235402 Rhizomucor Species 0.000 description 1
- 241000235527 Rhizopus Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 241000221662 Sclerotinia Species 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000172 allergic effect Effects 0.000 description 1
- 208000010668 atopic eczema Diseases 0.000 description 1
- OGBUMNBNEWYMNJ-UHFFFAOYSA-N batilol Chemical class CCCCCCCCCCCCCCCCCCOCC(O)CO OGBUMNBNEWYMNJ-UHFFFAOYSA-N 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- 235000019800 disodium phosphate Nutrition 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000011194 food seasoning agent Nutrition 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 235000015203 fruit juice Nutrition 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- VRIVJOXICYMTAG-IYEMJOQQSA-L iron(ii) gluconate Chemical compound [Fe+2].OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O.OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O VRIVJOXICYMTAG-IYEMJOQQSA-L 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 235000020124 milk-based beverage Nutrition 0.000 description 1
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- 150000003904 phospholipids Chemical class 0.000 description 1
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Classifications
-
- 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
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/58—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from fungi
- C12N9/62—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from fungi from Aspergillus
Abstract
The present invention aims to provide a technology for enhancing the soybean flavor of a soybean beverage. A method for producing a processed soybean beverage, comprising a step of allowing a protease derived from a filamentous fungus to act on a soybean beverage, wherein the processed soybean beverage obtained by the production method has an enhanced soybean flavor.
Description
Technical Field
The present invention relates to a method for producing a processed soybean beverage. More particularly, the present invention relates to processing techniques for enhancing the soy flavor of soy beverages.
Background
In recent years, the consumption of soybean beverages has been greatly increased due to various backgrounds such as health hot flashes, response to allergic problems, religious reasons, and increase in demand in houses such as expansion of infectious diseases.
The soybean beverage includes "soybean milk" having a soybean solid content of 8% by weight or more, which is produced from only soybean from which okara has been removed and water; a "prepared soybean milk" having a soybean solid content of 6% by weight or more and being easily drinkable by adding additives such as granulated sugar and rice oil, a "soybean milk beverage" having a soybean solid content of 2% by weight or more or 4% by weight and imparting a taste such as juice or coffee, and the like.
Several techniques for improving the flavor of soymilk have been reported so far. Examples of such a technique include: a method of obtaining soybean milk with reduced soybean odor by allowing a protease derived from a plant, an animal or a yeast to act on the soybean milk (patent document 1); and a method of obtaining soybean milk with reduced soybean odor by allowing aminopeptidase derived from Aspergillus oryzae to act on the soybean milk (patent document 2).
Prior art literature
Patent literature
Patent document 1: japanese patent application laid-open No. 2004-16215
Patent document 2: japanese patent laid-open publication No. 2005-105503
Disclosure of Invention
Technical problem to be solved by the invention
Heretofore, in the modification of the flavor of soybean beverages, processing for reducing soybean odor has been mainly performed. On the other hand, as the popularity of soybean beverages increases, the flavor preference becomes more diverse, and thus it is desired to further increase the range of palatability by increasing the original flavor of soybean instead.
Accordingly, an object of the present invention is to provide a technique for enhancing the soybean flavor of a soybean beverage.
Technical scheme for solving technical problems
The present inventors have found that the original flavor of soybean is enhanced by treating a soybean beverage with a protease derived from a filamentous fungus. That is, the present invention provides the following disclosed embodiments.
A method for producing a processed soybean beverage, comprising the step of allowing a protease derived from a filamentous fungus to act on the soybean beverage.
The method according to item 1, wherein the soybean beverage is soybean milk.
The method according to item 1 or 2, wherein the protease derived from a filamentous fungus is an acid protease.
The method according to any one of items 1 to 3, wherein the protease derived from a filamentous fungus is a protease derived from Aspergillus.
The method according to any one of items 1 to 4, wherein the protease derived from a filamentous fungus is a protease derived from Aspergillus oryzae.
The production method according to any one of items 1 to 5, wherein the processed soybean beverage has a viscosity of 8 mPas or more.
Item 7. A soy flavor enhancer for a soy beverage comprising a protease derived from a filamentous fungus.
Effects of the invention
According to the present invention, the soybean flavor of the soybean beverage can be enhanced by treating the soybean beverage with a protease derived from a filamentous fungus.
Detailed Description
1. Method for preparing processed soybean beverage
The method for producing a processed soybean beverage of the present invention is characterized by comprising a step of allowing a protease derived from a filamentous fungus to act on the soybean beverage. Hereinafter, a method for producing the processed soybean beverage will be described in detail.
The soybean beverage used in the present invention is a beverage using soybean as a material, and may be a liquid composition containing at least broken pieces of cotyledon parts of soybean seeds in a state of being dispersed in water. In the soybean beverage, the crushed material of the hypocotyl and/or the seed coat may be contained in a state of being dispersed in water, in addition to the crushed material of the cotyledon part.
Specific examples of the soybean beverage include a liquid for pulverizing and dispersing soybean in water; a liquid (i.e., soybean milk) obtained by removing okara from a liquid in which soybean is crushed and dispersed in water; and a liquid obtained by dissolving and/or dispersing a dry powder prepared from these liquids in water.
These soybean beverages may be used alone in an amount of 1 or in combination of two or more. Among these soybean beverages, soybean milk is preferred.
The amount of protein in the soybean beverage is not particularly limited, and examples thereof include 1g/100ml or more, preferably 2g/100ml or more, more preferably 3g/100ml or more, still more preferably 3.5g/100ml or more, and still more preferably 4g/100ml or more. The upper limit of the amount of protein in the soybean beverage is not particularly limited, and examples thereof include 8g/100ml or less, 6g/100ml or less, 5g/100ml or less, and 4.5g/100ml or less.
The soybean solid content in the soybean beverage is not particularly limited, and examples thereof include 2% by weight or more, preferably 4% by weight or more, and more preferably 8% by weight or more. The upper limit of the soybean solid content range in the soybean beverage is not particularly limited, and examples thereof include 16 wt% or less, 14 wt% or less, 12 wt% or less, or 10 wt% or less.
The soybean beverage used in the present invention may contain other components than soybean and water. Examples of the other components include quality improvers such as emulsifiers (sucrose fatty acid esters, phospholipids, monoglycerides, organic acid monoglycerides, sorbitan fatty acid esters, polyoxyethylene sorbitan fatty acid esters, polyglycerin fatty acid esters, propylene glycol fatty acid esters, etc.), pH adjusters (sodium carbonate, sodium hydrogencarbonate, calcium carbonate, etc.), colorants, flavors, seasonings (salt, granulated sugar (sucrose, etc.), and side materials (fruit juice, coffee, etc.).
The protease derived from a filamentous fungus used in the present invention is not particularly limited as long as it can obtain the desired effect of the present invention and can be used in foods. The protease derived from a filamentous fungus used in the present invention may be an acid protease, a neutral protease or an alkaline protease, but is preferably an acid protease from the viewpoint of further improving the soybean flavor-enhancing effect.
Specific examples of proteases derived from filamentous fungi include proteases derived from Aspergillus, mucor, neurospora, penicillium, rhizomucor, rhizopus, and Sclerotinia. These proteases derived from filamentous fungi may be used alone in an amount of 1 or in combination of two or more. Among these filamentous fungus proteases, proteases derived from Aspergillus are preferable from the viewpoint of further improving soybean flavor-enhancing effect.
Specific examples of proteases derived from Aspergillus include proteases derived from Aspergillus oryzae (Aspergillus oryzae), aspergillus niger (Aspergillus niger), aspergillus meli (Aspergillus melleus), aspergillus japonicus (Aspergillus japonicus), aspergillus awamori (Aspergillus awamori), aspergillus kawachii (Aspergillus kawachii), aspergillus sojae (Aspergillus sojae), aspergillus flavus (Aspergillus tamarii), aspergillus foetidus (Aspergillus foetidus), aspergillus fumigatus (Aspergillus fumigatus), aspergillus nidulans (Aspergillus nidulans), aspergillus aculeatus (Aspergillus aculeatus), aspergillus candidus (Aspergillus candidus), aspergillus flavus (Aspergillus flavus), aspergillus saitoi (Aspergillus saitoi), aspergillus drynarii (Aspergillus inuii), aspergillus glaucus (Aspergillus glaucus), aspergillus bluarius (Aspergillus caesiellus), aspergillus clavus (Aspergillus clavatus), aspergillus elbow (Aspergillus deflectus), aspergillus ferox (Aspergillus fischerianus), aspergillus parasiticus (Aspergillus parasiticus), aspergillus pencilis (Aspergillus penicilloides), aspergillus restrictus (Aspergillus restrictus), aspergillus polyrhizus (Aspergillus sydowii), aspergillus terreus (Aspergillus terreus), aspergillus kawachii (Aspergillus ustus), aspergillus versicolor (Aspergillus versicolor), and the like. These proteases derived from Aspergillus may be used alone in an amount of 1 or in combination of two or more. Among these proteases derived from Aspergillus, proteases derived from Aspergillus oryzae are preferable from the viewpoint of further improving soybean flavor enhancing effect. In addition, from the viewpoint of enhancing soybean flavor and suppressing the generation of bitterness, proteases derived from Aspergillus niger are preferable.
Proteases derived from filamentous fungi can be produced by known methods. For example, in the case of producing a protease derived from Aspergillus, it can be easily produced by a method of preparing a koji from Aspergillus and isolating the protease by a known method, a method using a genetic recombination technique, or the like. Further, commercially available proteases may be used as the proteases derived from the filamentous fungi. As commercially available proteases derived from filamentous fungi, there may be mentioned: protease HF "Amano"150SD (acid protease derived from Aspergillus oryzae), protease M "Amano" (acid protease derived from Aspergillus oryzae), acid protease UF "Amano" SD (acid protease derived from Aspergillus niger), protease A "Amano" (neutral protease derived from Aspergillus oryzae), protease A "Amano"2SD (neutral protease derived from Aspergillus oryzae) manufactured by Tianye enzyme Co., ltd; sumizyme MP (alkaline protease derived from Aspergillus oryzae), sumizyme FL-G (alkaline protease derived from Aspergillus oryzae) and the like of New Japanese chemical industry Co., ltd.
The amount of the protease derived from a filamentous fungus is not particularly limited, and examples of the amount per 1g of protein include 10U or more. From the viewpoint of further improving the effect of enhancing the soybean flavor of the soybean beverage, the amount of the protease derived from the filamentous fungus per 1g of the protein is preferably 100U or more, more preferably 150U or more, 1000U or more, 2000U or more, 3000U or more, or 4000U or more, more preferably 5000U or more, still more preferably 5500U or more. Further, from the viewpoint of enhancing the viscosity of the soybean beverage, the amount of the protease derived from the filamentous fungus per 1g of the protein is preferably 1000U or more, more preferably 1500U or more, still more preferably 3000U or more, still more preferably 4000U or more, still more preferably 5000U or more. The upper limit of the amount of the protease derived from a filamentous fungus is not particularly limited, and the amount of the protease per 1g of the protein may be 100000U or less or 50000U or less, for example. Further, from the viewpoint of suppressing the generation of bitter taste, the amount of protein per 1g may be preferably 10000U or less, more preferably 6000U or less, 5000U or less, 4000U or less, 3000U or less, 2000U or less, 1000U or less, 500U or less, 300U or 200U or less.
The amount of the protease derived from a filamentous fungus per 1g of the soybean solid content is, for example, 5U or more. The amount of the protease derived from a filamentous fungus per 1g of the soybean solid content is preferably 50U or more, more preferably 70U or more, 500U or more, 1000U or more, 1500U or more, or 2000U or more, and still more preferably 2300U or more, from the viewpoint of further improving the effect of enhancing the soybean flavor of the soybean beverage. Further, from the viewpoint of enhancing the viscosity of the soybean beverage, the amount of the protease derived from the filamentous fungus per 1g of the soybean solid content is preferably 500U or more, more preferably 680U or more, still more preferably 1300U or more, still more preferably 1800U or more, still more preferably 2300U or more. The upper limit of the amount of the protease derived from a filamentous fungus is not particularly limited, and examples of the amount of the protease per 1g of the soybean solid component include 50000U or less, 20000U or less, or 10000U or less. Further, from the viewpoint of suppressing the generation of bitterness, the amount of the soybean solid component per 1g may be preferably 4000U or less, more preferably 3000U or less, still more preferably 2700U or less, still more preferably 2300U or less, 1800U or less, 1400U or less, 900U or less, 450U or less, 230U or less, 140U or less, or 90U or less.
The protease activity derived from the filamentous fungus was measured by the Furline method using casein as a substrate. That is, in the present invention, the enzyme reaction was carried out by a conventional method using casein as a substrate, and the amount of enzyme that caused an increase in the color-developing substance of the forskolin solution corresponding to 1. Mu.g of tyrosine within 1 minute was defined as 1U of protease activity derived from a filamentous fungus.
In the step of allowing a protease derived from a filamentous fungus to act on a soybean beverage, a soybean beverage composition containing the soybean beverage and the protease derived from a filamentous fungus is prepared by adding the protease derived from a filamentous fungus to the soybean beverage, and a reaction for enhancing the flavor of soybean is performed by maintaining the soybean beverage composition in a heated state.
The reaction temperature in the step of allowing the protease derived from the filamentous fungus to act can be appropriately determined in consideration of the optimum temperature of the protease derived from the filamentous fungus, and examples thereof include 35 to 70 ℃, preferably 40 to 60 ℃, and more preferably 45 to 55 ℃.
In the step of allowing the protease derived from the filamentous fungus to act, other enzymes may be used in combination with the protease derived from the filamentous fungus as long as the effects of the present invention are not impaired.
The reaction time in the step of allowing the protease derived from the filamentous fungus to act is not particularly limited, and examples thereof include 30 minutes or longer, preferably 1 hour or longer. The upper limit of the reaction time range is not particularly limited, and examples thereof include 24 hours or less, 12 hours or less, 8 hours or less, 4 hours or less, or 2 hours or less.
The step of allowing the protease derived from the filamentous fungus to act may be completed by inactivating the protease derived from the filamentous fungus used. The method of inactivation is not particularly limited, and examples thereof include inactivation at a high temperature (for example, 85 to 100 ℃).
The soybean beverage composition after the step of acting the protease derived from the filamentous fungus is cooled to obtain the processed soybean beverage. The processed soybean beverage obtained by the production method of the present invention can be provided as a soybean beverage having enhanced soybean flavor.
The viscosity of the processed soybean beverage obtained by the production method of the present invention is, for example, 1 to 70 mPas, preferably 2 to 60 mPas. The processed soybean beverage obtained by the production method of the present invention may have a viscosity increased according to the amount of the protease derived from the filamentous fungus, although the processed soybean beverage is treated with the protease derived from the filamentous fungus. The lower limit of the viscosity range in the case of viscosity enhancement is, for example, 8mpa·s or more, preferably 10mpa·s or more, more preferably 20mpa·s or more, still more preferably 30mpa·s or more, still more preferably 40mpa·s or more, still more preferably 50mpa·s or more. The viscosity was measured at 20℃at 1000rpm as the rotational speed of the rotor in the ball rotary viscometer.
The processed soybean beverage thus obtained may be prepared by a drying process as a processed soybean composition which is a solid to obtain a soybean beverage having an enhanced soybean flavor when dissolved and/or dispersed in water. The method of drying is not particularly limited, and examples thereof include freeze drying, vacuum drying, spray drying, and the like. The shape of the processed soybean composition as a solid may be powder, granule, or the like.
2. Soybean flavor enhancer for soybean beverage
As described above, proteases derived from filamentous fungi can enhance the soybean flavor of soybean beverages. Thus, the present invention also provides a soy-like enhancer for soy beverages comprising a protease derived from a filamentous fungus.
The type and amount of the materials and components used in the soybean-like reinforcing agent for soybean beverage are as shown in column "1. Method for producing processed soybean beverage".
Examples
The present invention will be specifically described with reference to the following examples, which are not to be construed as limiting the invention.
(1) Material
Soymilk: the soybean milk has no adjustment, the protein concentration is 4.15g/100mL, and the solid content of the soybean is more than 8 wt%
PR-HF150SD (protease HF "Amano"150SD, tianye Co., ltd.): protease derived from the filamentous fungus Aspergillus oryzae
PR-UFSD (acid protease UF "Amano" SD, tianye enzyme Co., ltd.): protease derived from Aspergillus niger
TH-PC10F (Thermoase PC10F, tianye enzyme Co., ltd.): protease derived from Geobacillus stearothermophilus (Geobacillus stearothermophilus)
LC-Y120 (laccase Y120, wild enzyme): laccase derived from Trametes (Trametes)
(2) Enzyme activity value assay
(2-1) protease Activity assay
After heating 5mL of 0.6% (w/v) casein solution (0.05 mol/L sodium hydrogen phosphate, pH8.0[ TH-PC10F ]) or 0.6% (v/w) casein solution (0.7% (v/w) lactic acid, pH3.0[ PR-HF150SD and PR-UFSD ]) at 37℃for 10 minutes, 1mL of a sample solution containing a protease was added and immediately mixed by shaking. After the liquid was left at 37℃for 10 minutes, 5mL of a trichloroacetic acid test solution (containing 1.8 (w/v)% trichloroacetic acid, 1.8% (w/v) sodium acetate and 0.33mol/L acetic acid) was added, and mixed with shaking, and left at 37℃again for 30 minutes, followed by filtration. The initial 3mL of the filtrate was removed, 2mL of the subsequent filtrate was measured, 5mL of a sodium carbonate solution (0.55 mol/L) and 1mL of Fu Lin Shiye (1.fwdarw.3) were added thereto, and the mixture was thoroughly mixed by shaking and left at 37℃for 30 minutes. The absorbance AT AT 660nm was measured with respect to the liquid (enzyme reaction liquid) by using water as a control.
Further, 1mL of a sample solution containing a protease was measured, 5mL of a trichloroacetic acid sample solution (containing 1.8 (w/v)% trichloroacetic acid, 1.8 (w/v)% sodium acetate and 0.33mol/L acetic acid) was added thereto, and after mixing by shaking, 5mL of the sample solution was added thereto, mixing was immediately performed by shaking, and the mixture was left to stand at 37℃for 30 minutes, and the absorbance AB was measured for a liquid (blank) operated in the same manner as the enzyme reaction liquid.
The amount of enzyme that caused an increase in the color development substance of the forskolin test solution corresponding to 1. Mu.g of tyrosine within 1 minute was set to 1 unit (1U).
1mg/mL of tyrosine standard stock solution (0.2 mol/L hydrochloric acid) 1mL, 2mL, 3mL and 4mL are measured, and 0.2mol/L hydrochloric acid test solution is added respectively to prepare 100mL. 2mL of each liquid was weighed, 5mL of a sodium carbonate solution (0.55 mol/L) and 1mL of Fu Lin Shiye (1.fwdarw.3) were added thereto, and immediately mixed by shaking, and the mixture was left at 37℃for 30 minutes. For these liquids, absorbance A1, A2, A3 and A4 at 660nm was measured by taking 2mL of a hydrochloric acid sample solution of 0.2mol/L as a control in the same manner as described above. The absorbance A1, A2, A3 and A4 was taken along the vertical axis, the tyrosine amount (. Mu.g) in 2mL of each liquid was taken along the horizontal axis, and a standard curve was prepared to determine the tyrosine amount (. Mu.g) relative to the absorbance difference 1.
[ number 1]
Protease Activity (U/g, U/mL) = (AT-AB) ×F×11/2×1/10×1/M
AT: absorbance of enzyme reaction solution
AB: absorbance of blank
F: tyrosine amount (. Mu.g) at 1 absorbance difference obtained from tyrosine standard curve
11/2: conversion coefficient to total liquid amount after stopping reaction
1/10: conversion coefficient to reaction time every 1 minute
M: sample amount (g or mL) in 1mL of sample solution
(1-1) laccase Activity value assay
The enzymatic activity of laccase was measured by the method described below using 2,2 '-diaza-bis [ 3-ethylbenzothiazoline sulfonate ] (2, 2' -azino-di- [3-ethylbenzthiazoline sulfonate ]) (ABTS, manufactured by Boehringer Mannheim Co.) as a substrate.
ABTS was dissolved in 25mM citric acid buffer (ph 3.2) at a concentration of 1.0mg/ml as a substrate solution. 3.0ml of the substrate solution was placed in a test tube (cuvette), and after preheating at 25℃0.1ml of enzyme solution was added, stirred, incubated at 25℃and absorbance at 405nm after 1 minute and after 3 minutes was measured. Under this condition, the amount of enzyme that increases absorbance at 405nm by 1.0OD in 1 minute was defined as 1 unit (U).
Test example
The enzyme was dissolved in water to prepare an enzyme solution. In the non-adjusted soybean milk, the enzyme solution was added so that the enzyme shown in tables 1 and 2 became the final concentration shown, and reacted in a hot water bath at 50℃for 90 minutes while stirring. Then, the enzyme was deactivated in a hot water bath at 90℃and cooled to room temperature. The following evaluation was performed on the obtained processed soybean milk. The results are shown in tables 1 and 2.
(evaluation of soybean flavor enhancement)
The 7-person inspector drunk the processed soybean milk and evaluated the soybean flavor-enhancing effect based on the flavor of the raw material soybean milk based on the following criteria.
And (3) the following materials: greatly enhancing soybean flavor
And (2) the following steps: the flavor of the soybean is enhanced to a certain extent
Delta: slightly enhanced soy flavor
X: soybean flavor is not enhanced
(bitter taste production inhibitory Property)
The 7-person inspector drinks the processed soybean milk, and evaluates the effect of suppressing the generation of bitter taste by the protease treatment based on the flavor of the soybean milk as a raw material based on the following criteria.
And (3) the following materials: has no bitter taste and can inhibit bitter taste.
And (2) the following steps: the bitter taste is reduced and the generation of bitter taste is well suppressed.
Delta: the bitter taste is generated to some extent, but the bitter taste is inhibited to such an extent that the bitter taste does not interfere with drinking.
X: the bitter taste is rather strong, and the generation of bitter taste is not suppressed to the extent that the drinking is hindered.
(viscosity)
Using a ball rotary viscometer (EMS-1000, 1000rpm, ball probe, manufactured by Kyoto electronic industries Co., ltd.)) The viscosity of the processed soymilk was measured at 20 ℃.
TABLE 1
TABLE 2
As is clear from tables 1 and 2, soybean original flavor was enhanced by treating soybean milk with protease derived from filamentous fungi (examples 1 to 5). The viscosity of the soybean milk was also confirmed to be increased by the amount of protease derived from the filamentous fungi, and a thick texture was obtained in addition to the original flavor of soybean (examples 2 to 5). In the case of treatment with a protease derived from bacteria, the viscosity of soybean milk was sometimes increased (in particular, comparative example 3), but the effect of enhancing soybean flavor was not confirmed. That is, it is found that the soybean flavor enhancement effect confirmed in examples is generated irrespective of the thickening of soybean milk, that is, is a characteristic effect exerted by proteases derived from filamentous fungi. Further, the protease treatment showed a tendency to cause bitter taste as the amount of protease was increased, and as a result, even in the case of the protease treatment using a bacterium-derived protease, bitter taste which could not be drunk was caused even in the case of the protease treatment using a filamentous bacterium-derived protease, and even in the case of the protease treatment using a certain amount, bitter taste was prevented from being produced to such an extent that drinking was not hindered. Further, by controlling the amount of protease derived from filamentous fungi, the generation of bitterness can be further suppressed, and the soybean flavor enhancing effect and the thickening effect of soybean milk can be well balanced, whereby processed soybean milk having higher palatability can be produced (examples 1 to 4, particularly examples 1, 3 and 4).
Claims (7)
1. A method for producing a processed soybean beverage, characterized by comprising a step of allowing a protease derived from a filamentous fungus to act on the soybean beverage.
2. The manufacturing method according to claim 1, wherein the soybean beverage is soybean milk.
3. The production method according to claim 1 or 2, wherein the protease derived from a filamentous fungus is an acid protease.
4. The method according to any one of claims 1 to 3, wherein the protease derived from a filamentous fungus is a protease derived from Aspergillus.
5. The method according to any one of claims 1 to 4, wherein the protease derived from a filamentous fungus is a protease derived from Aspergillus oryzae.
6. The method according to any one of claims 1 to 5, wherein the processed soybean beverage has a viscosity of 8mpa·s or more.
7. A soy flavor enhancer for a soy beverage, wherein the soy flavor enhancer comprises a protease derived from a filamentous fungus.
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