Emulsibility-improving agent and foamability-improving agent for liquid composition containing vegetable protein and oil / fat

Abstract

The purpose of the present invention is to provide a processing technique with which it is possible to improve the emulsibility and / or foamability of a liquid composition containing a vegetable protein. If a liquid composition containing a vegetable protein and an oil or fat is treated with a protein deamidase and a lipase, the emulsibility and / or foamability of the liquid composition can be improved.

Description

Technical Field

[0001] The present invention relates to a processing technology capable of improving the emulsification and / or foaming properties of a liquid composition containing vegetable protein. Specifically, the present invention relates to an emulsification enhancer or foaming enhancer for a liquid composition containing vegetable protein and fat, a method for improving the emulsification and foaming properties of a liquid composition containing vegetable protein and fat, a method for producing an emulsion of a liquid composition containing vegetable protein and fat, and an emulsion of a liquid composition containing vegetable protein and fat. Background Art

[0002] In recent years, the milk alternative market and the meat alternative market have expanded in the food market against the backdrop of sustainability and health intentions. Among them, plant-based beverages, which are substitutes for animal milk, are one of the important categories in the milk alternative market and continue to attract attention.

[0003] Since plant-based beverages have a fundamentally different composition of ingredients from milk, they must be processed to approximate the properties of animal milk. Therefore, various studies have been conducted on processing techniques for plant-based beverages.

[0004] As an example of the characteristics of animal milk that plant-based beverages imitate, emulsification characteristics can be cited. Generally, beverages containing plant proteins are formulated with emulsifiers in order to maintain their emulsified state well. As such emulsifiers, glycerol fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, sucrose fatty acid esters, polyglycerol fatty acid esters, gum arabic, lecithin, etc. are used as described in Patent Document 1. On the other hand, processing techniques for emulsification treatment using methods other than emulsifiers have also been studied. For example, in Patent Document 2, a method using a plant-based liquid nutrition composition that meets specified requirements is described as a technique for producing a plant-based liquid nutrition composition that has satisfactory emulsification stability even without the addition of milk protein.

[0005] Another example of the characteristics of animal milk that plant-based beverages imitate is foaming property. For example, Patent Document 3 describes that good foaming property can be imparted to soy milk by heating low-fat soy milk.

[0006] Prior art literature Patent Literature Patent Document 1: Japanese Patent Application Publication No. 2021-176284 Patent Document 2: Japanese Patent Application Publication No. 2021-52655 Patent Document 3: International Publication No. 2014 / 092157 Summary of the invention Problems to be solved by the invention Plant-based beverages are particularly concerned by users with high health awareness, so there is a trend of expecting plant-based beverages that do not use emulsifiers as additives. However, at present, in order to give excellent emulsification to plant-based beverages, emulsifiers have to be relied on. In addition, as shown in the record of Patent Document 2, a method of using a plant protein raw material that meets the specified requirements has also been proposed, but as the requirements of the raw material, not only the protein content, but also the NSI, molecular weight distribution and gelation characteristics all need to meet the specified range. In view of the fact that such strict raw materials are not easy to obtain, it is desired to use a more easily available material to improve the emulsification method.

[0007] In addition, the foaming property of plant-based beverages has not been fully studied. Although the method described in Patent Document 3 is proposed, this method can only be applied to soy milk. In addition, it is generally difficult to foam plant-based beverages. However, in view of the expansion of the milk substitute market in the future, a method for improving the foaming property that can be applied to other types of plant-based beverages is desired.

[0008] Therefore, an object of the present invention is to provide a processing technology capable of improving the emulsification and / or foaming properties of a liquid composition containing a vegetable protein.

[0009] Technical solutions for solving technical problems The present inventors have found that emulsification and / or foaming properties can be improved by adding fat to a liquid composition containing vegetable protein and treating it with protein deamidase and lipase. It has also been found that emulsification and / or foaming properties can be further improved by treating it with protease in addition to protein deamidase and lipase. The present invention has been completed through further research based on these findings.

[0010] That is, the present invention provides the following aspects.

[0011] Item 1. An agent for improving the emulsification properties of a liquid composition containing a vegetable protein and an oil, the agent comprising a protein deamidase and a lipase.

[0012] Item 2. The emulsification enhancer according to Item 1, wherein the amount of the lipase is 1 U or more per 1 U of the protein deamidase.

[0013] Item 3. The emulsification enhancer according to Item 1 or 2, further comprising a protease.

[0014] Item 4. A foaming enhancer for a liquid composition comprising vegetable protein and fat, the foaming enhancer comprising a protein deamidase and a lipase.

[0015] Item 5. The foaming enhancer according to Item 4, further comprising a protease.

[0016] Item 6. The foaming property enhancer according to Item 5, wherein the amount of the protease is 1 U or more per 1 U of the protein deamidase.

[0017] Item 7. A method for improving the emulsification property of a liquid composition containing vegetable protein and oil, comprising an enzyme treatment step of allowing a protein deamidase and a lipase to act on the liquid composition.

[0018] Item 8. The method according to Item 7, wherein 0.05 U or more of the protein deamidase is used per 1 g of the vegetable protein.

[0019] Item 9. The method according to Item 7 or 8, wherein 1 U or more of the lipase is used per 1 g of the fat or oil.

[0020] Item 10. A method for improving the foaming property of a liquid composition containing vegetable protein and oil, the method comprising an enzyme treatment step of allowing a protein deamidase and a lipase to act on the liquid composition.

[0021] Item 11. A method for producing an emulsion of a liquid composition containing vegetable protein and oil, the method comprising an enzyme treatment step of treating the liquid composition with a protein deamidase and a lipase.

[0022] Item 12. The production method according to Item 11, wherein a protease is further used in the enzyme treatment step.

[0023] Item 13. A food or beverage which is an emulsion of a liquid composition containing a vegetable protein and fat obtained by the production method of Item 11 or 12.

[0024] Item 14. Use of an enzyme preparation comprising a protein deamidase and a lipase for improving the emulsification and / or foaming properties of a liquid composition comprising a vegetable protein and an oil.

[0025] Item 15. Use of an enzyme preparation comprising a protein deamidase and a lipase as an emulsifying agent and / or a foaming agent for a liquid composition comprising a vegetable protein and an oil.

[0026] Effects of the Invention According to the present invention, there is provided a processing technology capable of improving the emulsification and / or foaming properties of a liquid composition containing a vegetable protein. DETAILED DESCRIPTION

[0027] 1. Emulsification enhancer and foaming enhancer The emulsification enhancer and foaming enhancer of the liquid composition containing vegetable protein and fat of the present invention are characterized by containing protein deamidase and lipase. The emulsification enhancer and foaming enhancer of the present invention are described in detail below.

[0028] 1-1. Specified enzymes The emulsification enhancer and foaming enhancer of the present invention contain protein deamidase and lipase as active ingredients, and preferably further contain protease from the viewpoint of further improving the emulsification enhancer effect and foaming enhancer effect.

[0029] 1-1-1. Protein deamidase The protein deamidase used in the present invention is not particularly limited in type or origin as long as it is an enzyme that decomposes the amide group-containing side chain of a protein without cleavage of peptide bonds or cross-linking of proteins.

[0030] Examples of protein deamidase include enzymes that deamidate glutamine residues in proteins to convert them into glutamate (e.g., protein glutaminase) and enzymes that deamidate asparagine residues in proteins to convert them into aspartic acid (e.g., protein asparaginase).

[0031] As a more specific example of protein deamidase, there can be mentioned: protein deamidase derived from Chryseobacterium, Flavobacterium, Empedobacter, Sphingobacterium, Aureobacterium, Myroides, Luteimicrobium, Agromyces, Microbacterium, or Leifsonia. These protein deamidase are well known, for example, JP2000-50887A, JP2001-218590A, WO2006 / 075772A1, WO2015 / 133590, etc. These protein deamidase can be used alone or in combination.

[0032] Among these protein deaminases, from the viewpoint of further improving the emulsification and / or foaming properties improving effects, preferably, protein glutaminase is used, more preferably, protein glutaminase derived from the genus Chryseobacterium is used, and further preferably, protein glutaminase derived from the species Chryseobacterium proteolyticum is used.

[0033] Protein deamidase can be prepared from the culture solution of the microorganism that is the source of the above-mentioned protein deamidase. As a specific preparation method, a method for recovering protein deamidase from the culture solution or bacterial cell of the above-mentioned microorganism can be cited. For example, in the case of using a protein deamidase secretory microorganism, the bacterial cell can be recovered from the culture solution by filtering, centrifugation, etc. in advance as needed, and the enzyme is separated and / or purified. In addition, in the case of using a protein deamidase non-secreting microorganism, the bacterial cell can be recovered from the culture solution in advance as needed, and the enzyme is exposed by crushing the bacterial cell by pressure treatment, ultrasonic treatment, etc., and then the enzyme is separated and / or purified. As an enzyme separation and / or purification method, a known protein separation and / or purification method can be used without particular limitation, for example, centrifugation, UF concentration method, salting-out method, various chromatography methods using ion exchange resins, etc. can be cited. The enzyme obtained by separation and / or purification can be powdered by drying methods such as freeze drying and reduced pressure drying, and in addition, it can also be powdered using appropriate excipients and / or drying aids in the drying method. Alternatively, the isolated and / or purified enzyme may be liquefied by adding appropriate additives and sterilizing by filtration.

[0034] The amount of the protein deamidase contained in the emulsification and foaming agents of the present invention is not particularly limited as long as the effect of the protein deamidase can be effectively obtained when the emulsification and foaming agents of the present invention are used, and examples thereof include 0.1 to 5000 U / g.

[0035] Regarding the activity of protein deamidase, benzyloxycarbonyl-L-glutamine glycine (Z-Gln-Gly) was used as substrate and 1 μ The amount of enzyme per mol of ammonia is taken as 1 unit (1U).

[0036] 1-1-2. Lipase The lipase used in the present invention is not particularly limited in type or origin as long as it is an enzyme that hydrolyzes triglycerides.

[0037] Examples of lipases include lipases derived from the genus Rhizopus, the genus Aspergillus, the genus Mucor, the genus Rhizomucor, the genus Thermomyces, the genus Pseudomonas, the genus Geotrichum, the genus Penicillium, and the genus Candida.

[0038] As lipases derived from the genus Rhizopus, for example, lipases derived from Rhizopus delemar, Rhizopus oryzae, Rhizopus arrhizus, Rhizopus niveus, Rhizopus javanicus, etc. can be cited. As lipases derived from the genus Aspergillus, for example, lipases derived from Aspergillus niger, etc. can be cited. As lipases derived from the genus Mucor, for example, lipases derived from Mucor javanicus, Mucor miehei, etc. can be cited. As lipases derived from the genus Rhizomucor, for example, lipases derived from Rhizomucor miehei, etc. can be cited. As lipases derived from the genus Thermomyces, for example, lipases derived from Thermomyces lanuginosus, etc. can be cited.

[0039] These lipases may be used alone or in combination. Among these lipases, from the viewpoint of further improving the emulsification and / or foaming properties, preferably, a lipase derived from Rhizopus is used, and more preferably, a lipase derived from Rhizopus oryzae is used.

[0040] In addition, as the lipase used in the present invention, from the viewpoint of further improving the emulsification effect and / or the foaming effect, preferably, there can be mentioned a lipase having a 1 ( α ) bit, 2 ( β ) bit, 3 ( γ ) at one or two positions (specifically, position 1, position 2, position 3, position 1 and position 2, position 1 and position 3, or position 2 and position 3), more preferably, an enzyme having the effect of hydrolyzing the ester bonds at positions 1 and 3 of triglyceride, and still more preferably, an enzyme having the effect of site-specifically hydrolyzing the ester bonds at positions 1 and 3 of triglyceride.

[0041] The amount of lipase contained in the emulsification enhancer and foaming enhancer of the present invention is not particularly limited as long as the effect of lipase can be effectively obtained when the emulsification enhancer and foaming enhancer of the present invention is used, and examples thereof include 1 to 1,500,000 U / g.

[0042] In the emulsification enhancer and foaming enhancer of the present invention, the amount of lipase per 1U of protein deamidase may be, for example, 0.1U or more, 0.3U or more, 0.5U or more, or 1U or more. From the viewpoint of further improving the emulsification enhancer effect and / or foaming enhancer effect, preferably, it may be 2U or more, 2.5U or more, 3U or more, 4U or more, 7U or more, or 10U or more. More preferably, it may be 12U or more, 13U or more, or 14U or more. The upper limit of the range of the amount of lipase per 1U of protein deamidase is not particularly limited, and for example, it may be 1000U or less, or 600U or less. From the viewpoint of further improving the emulsification improving effect and / or the foaming improving effect, it may preferably be 300U or less, 200U or less, 160U or less, 150U or less, 140U or less, or 120U or less. It may more preferably be 100U or less, 90U or less, 80U or less, or 75U or less. It may further preferably be 70U or less, 60U or less, 50U or less, 40U or less, 30U or less, or 20U or less.

[0043] Regarding the activity of lipase, using olive oil as substrate, 1 μ The amount of enzyme that increased 1 mol of free fatty acid was defined as 1 unit (1 U).

[0044] 1-1-3. Protease The protease used in the present invention is not particularly limited in type and origin, etc. It is preferable to use endopeptidase as the protease.

[0045] Examples of proteases include proteases derived from filamentous fungi and proteases derived from bacteria.

[0046] The protease derived from filamentous fungi is not particularly limited, and examples thereof include proteases derived from the genera Aspergillus, Rhizopus, Mucor, Neurospora, Penicillium, Rhizomucor, and Sclerotinia.

[0047] Specific examples of the protease derived from the genus Aspergillus include proteases derived from Aspergillus oryzae, Aspergillus niger, Aspergillus melleus, Aspergillus japonicus, Aspergillus awamori, Aspergillus kawachii, Aspergillus sojae, Aspergillus tamarii, Aspergillus foetidus, Aspergillus fumigatus, Aspergillus nidulans, Aspergillus aculeatus, Aspergillus candidus, Aspergillus flavus, Aspergillus saito, and Aspergillus saito. saitoi, Aspergillus inuii, Aspergillus glaucus, Aspergillus caesiellus, Aspergillus clavatus, Aspergillus deflectus, Aspergillus fischerianus, Aspergillus parasiticus, Aspergillus penicilloides, Aspergillus restrictus, Aspergillus sydowii, Aspergillus terreus, Aspergillus ustus, and Aspergillus versicolor.

[0048] Specific examples of the protease derived from the genus Rhizopus include proteases derived from Rhizopus chinensis, Rhizopus delemar, Rhizopus niveus, Rhizopus oryzae, and the like.

[0049] Examples of proteases derived from bacteria include proteases derived from the genus Bacillus and the genus Geobacillus.

[0050] Specific examples of the protease derived from the genus Bacillus (or Geobacillus) include proteases derived from Bacillus amyloliquefaciens, Bacillus cereus, Bacillus clausii, Bacillus intermedius, Bacillus lentus, Bacillus licheniformis, Bacillus stearothermophilus, Bacillus subtilis and Bacillus thermoproteolyticus, and proteases derived from Geobacillus subtilis thereof.

[0051] In the present invention, the above-mentioned proteases may be used alone or in combination. Among the above-mentioned proteases, from the viewpoint of further improving the emulsification and / or foaming properties, preferably, proteases derived from filamentous fungi are used, more preferably, proteases derived from Aspergillus are used, and even more preferably, proteases derived from Aspergillus oryzae are used.

[0052] The amount of the protease contained in the emulsification enhancer and foaming enhancer of the present invention is not particularly limited as long as the effect of the protease can be effectively obtained when the emulsification enhancer and foaming enhancer of the present invention is used, and for example, it is 1 to 500,000 U / g.

[0053] When the emulsification improving agent and foaming improving agent of the present invention contain a protease, the amount of the protease per 1U of the protein deamidase may be, for example, 0.1U or more, 0.3U or more, 0.5U or more, or 1U or more. From the viewpoint of further improving the emulsification improving effect and / or the foaming improving effect, it is preferably 2U or more, more preferably 3U or more, and even more preferably 3.5U or more. The upper limit of the range of the amount of the protease per 1U of the protein deamidase is not particularly limited, but may be, for example, 150U or less. From the viewpoint of further improving the emulsification improving effect and / or the foaming improving effect, it is preferably 100U or less, 75U or less, or 50U or less, more preferably 25U or less, or 15U or less, more preferably 10U or less, more preferably 5U or less, and even more preferably 4U or less.

[0054] Regarding the activity of the protease, using casein as substrate, it will produce the equivalent of 1 tyrosine in 1 minute. μ The amount of enzyme added to produce the color-developing substance in g of Folin test solution is taken as 1 unit (1U).

[0055] 1-2. Other ingredients The emulsification improver and foaming improver of the present invention can be prepared as an enzyme preparation containing the above-mentioned enzymes, and can contain other components in addition to the above-mentioned enzymes to the extent that the effects of the present invention are not affected. As other components, enzymes other than the above-mentioned enzymes and / or bases and / or additives used in common enzyme preparations can be cited. These other components can be appropriately determined according to the use of the emulsification improver and foaming improver and the formulation form.

[0056] 1-3. Preparation form The emulsification enhancer and foaming enhancer of the present invention may be in the form of a liquid or solid formulation.

[0057] 1-4. Application The emulsification enhancer and foaming enhancer of the present invention are used to enhance the emulsification and foaming properties of a liquid composition containing vegetable protein and fat, respectively. Specifically, the emulsification enhancer and foaming enhancer of the present invention can be used in the enzyme treatment step in the following method.

[0058] A method for improving the emulsification of a liquid composition containing vegetable protein and oil, comprising an enzyme treatment step of allowing protein deamidase and lipase to act on the liquid composition; A method for improving the foaming property of a liquid composition containing vegetable protein and fat, comprising an enzyme treatment step of allowing protein deamidase and lipase to act on the liquid composition; or A method for producing an emulsion of a liquid composition containing vegetable protein and fat, comprising an enzyme treatment step of treating the liquid composition containing vegetable protein and fat with protein deamidase and lipase.

[0059] Details of the liquid composition and enzyme treatment step in the above method are described in detail in "2. Method for improving the emulsification property of a liquid composition containing vegetable protein and fat, and method for improving the foaming property of a liquid composition containing vegetable protein and fat" described later.

[0060] 2. A method for improving the emulsification of a liquid composition containing vegetable protein and oil, and a method for improving the emulsification of a liquid composition containing vegetable protein and oil Method for improving foaming property of liquid composition of physical protein and fat The method for improving the emulsification of a liquid composition containing vegetable protein and fat, and the method for improving the foaming of a liquid composition containing vegetable protein and fat of the present invention are characterized in that both include an enzyme treatment step of allowing protein deamidase and lipase to act on the liquid composition containing vegetable protein and fat. These methods of the present invention are described in detail below.

[0061] 2-1. Liquid composition containing vegetable protein and oil The liquid composition to be treated with the enzyme in the method of the present invention contains vegetable protein and oil.

[0062] 2-1-1. Plant protein Plant proteins are not particularly limited, and examples thereof include proteins from beans such as soybeans, peas, lupines, broad beans, chickpeas, mung beans, and kidney beans; proteins from cereals such as oats, barley, wheat, rye, rice, buckwheat, barnyard grass, chestnuts, teff, quinoa, and corn; proteins from seeds such as canarygrass seeds, flax seeds, almonds, cashews, hazelnuts, pecans, macadamia nuts, pistachios, walnuts, Brazil nuts, peanuts, coconuts, hemp (industrial hemp), pili nuts, chestnuts, sesame seeds, and pine nuts, etc. In addition, these proteins may be in the form of natural products; in the form of chemical partial decomposition of proteins using acids, alkalis, etc.; in the form of chemically modified proteins using various reagents; and in the form of synthetic proteins. In addition, when these vegetable proteins are in the form of natural products, the vegetable proteins may be vegetable substances included as a material for a liquid composition along with the use of the source plant material (specifically, the above-mentioned beans, cereals, seeds, etc.), or may be substances included as a material for a liquid composition by using a processed material in which the protein content of the plant material is increased to an arbitrary degree (for example, the protein content is 20 to 100% by weight). The form of the processed material is not particularly limited, and examples thereof include powder, slurry, liquid, etc.

[0063] In the present invention, the above-mentioned vegetable proteins may be used alone or in combination. From the viewpoint of further improving emulsification and / or foaming properties, preferably, bean proteins and seed proteins are used, more preferably, proteins of soybean, pea, lupine, broad bean, chickpea, mung bean, kidney bean, canary grass seed, flax seed, almond, cashew, hazelnut, pecan kernel, macadamia nut, pistachio, walnut, Brazil nut, peanut, coconut, hemp (industrial hemp), pili nut, chestnut, sesame, pine nut are used, and more preferably, proteins of pea and almond are used.

[0064] The content of the vegetable protein in the liquid composition is not particularly limited, and may be, for example, 0.1 to 20% by weight, preferably 0.2 to 10% by weight, and more preferably 0.35 to 4% by weight.

[0065] The content of the plant material or the processed material in the liquid composition (wherein, when these materials contain water, the content does not include the water content) is, for example, 0.1 to 50% by weight, preferably 0.2 to 35% by weight, and more preferably 0.4 to 20% by weight.

[0066] 2-1-2. Grease As fats and oils, there are no particular limitations, and vegetable fats and oils are typically used. As vegetable fats and oils, fats and oils of beans such as soybeans, peas, lupines, broad beans, chickpeas, mung beans, and kidney beans can be cited; fats and oils of cereals such as oats, barley, wheat, rye, rice, buckwheat, barnyard grass, chestnuts, teff, quinoa, and corn; and fats and oils of seeds such as rapeseed, sunflower seeds, camellia seeds, safflower seeds, cotton seeds, oil palm fruits, coconut fruits, cocoa, avocados, olives, canary grass seeds, flax seeds, shea nuts, almonds, cashews, hazelnuts, pecan kernels, macadamia nuts, pistachios, walnuts, Brazil nuts, peanuts, coconuts, hemp (industrial hemp), pili nuts, chestnuts, sesame seeds, and pine nuts can be cited. In addition, as fats and oils, animal fats and oils are also allowed to be used. As animal fats and oils, fish oil, lard, beef tallow, and butter fat can be cited. In addition, these oils and fats may be in the form of natural products; hydrogenated oils; fractionated oils; and synthetic oils and fats obtained by transesterification. In addition, when the oils and fats are vegetable oils and fats and are in the form of natural products, the vegetable oils and fats may be vegetable substances included in the liquid composition along with the plant materials from which the oils and fats are used (specifically, the beans, cereals, seeds, etc. mentioned above), or may be substances included in the liquid composition by using processed materials in which the oil and fat content of the plant materials is increased to any degree (for example, the oil and fat content is 40 to 100% by weight). The form of the processed material is not particularly limited, and examples thereof include powder, slurry (i.e., butter form), liquid, etc.

[0067] In the present invention, the above-mentioned oils and fats may be used alone or in combination. From the viewpoint of further improving emulsification and / or foaming properties, preferably, vegetable oils and fats may be used, more preferably, oils and fats of soybean, pea, rice, corn, rapeseed, sunflower seed, safflower seed, cottonseed, oil palm fruit, coconut fruit, cocoa fruit, almond, cashew nut, and peanut may be used, and further preferably, oils and fats of pea, rapeseed, sunflower seed, and almond may be used.

[0068] When the fat is a vegetable fat, the plant from which the vegetable fat is derived may be the same as or different from the plant from which the vegetable protein is derived, or a combination thereof may be used.

[0069] The content of fat in the liquid composition is not particularly limited and can be appropriately determined according to the type of the target emulsified composition and / or foamable composition. For example, it can be 0.1 to 25% by weight. From the viewpoint of further improving the emulsification and / or foaming properties, it is preferably 1 to 22% by weight, more preferably 3 to 20% by weight, further preferably 4.8 to 18% by weight, and further preferably 10 to 17% by weight or 13 to 16% by weight.

[0070] The ratio of the vegetable protein content to the fat content is determined based on the above-mentioned contents. The fat content relative to 1 part by weight of vegetable protein is preferably 0.1 to 60 parts by weight, more preferably 0.5 to 50 parts by weight, further preferably 10 to 47 parts by weight, further preferably 20 to 45 parts by weight, or 35 to 40 parts by weight.

[0071] In addition, when the oil is a vegetable oil, the content of the above-mentioned plant material or the above-mentioned processed material contained in the liquid composition (wherein, when these materials contain water, the content does not include water) can be, for example, 0.1 to 40% by weight, preferably 1 to 35% by weight, more preferably 2.5 to 30% by weight, further preferably 5 to 25% by weight, further preferably 10 to 20% by weight, or 13 to 17% by weight.

[0072] 2-1-3. Other ingredients In addition to the above-mentioned ingredients and water, the liquid composition may also contain seasonings, emulsifiers, gelling agents, etc. as appropriate. As seasonings, granulated sugar, salt, etc. may be mentioned, and granulated sugar may be preferably mentioned. When the liquid composition contains granulated sugar, the content of granulated sugar may be, for example, 0.1 to 1.5% by weight, preferably 0.3 to 1.2% by weight, and more preferably 0.5 to 1% by weight. As emulsifiers, substances used as emulsifiers in alternative beverages for animal milk may be mentioned, and specifically, glycerol fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, sucrose fatty acid esters, polyglycerol fatty acid esters, gum arabic, lecithin, etc. may be mentioned.

[0073] The emulsification enhancer and foaming enhancer of the present invention are each excellent in the emulsification enhancer effect and foaming enhancer effect of the liquid composition. Therefore, as a preferred example of the liquid composition, it is preferred that no emulsifier (particularly lecithin) and / or gelling agent is contained, and it is more preferred that no emulsifier (particularly lecithin) and gelling agent are contained.

[0074] 2-1-4. Form The liquid composition is limited to the liquid state, and its form is not particularly limited. For example, the liquid composition may be in the form of an emulsified composition, or may be in the form of a non-emulsified composition. In addition, the liquid composition may be in the form of a foaming composition, or may be in the form of a non-foaming composition. The emulsification improver and the foaming improver of the present invention each have excellent emulsification and foaming effects on the liquid composition, so as a preferred example of the form of the liquid composition, preferably, a non-emulsified composition and / or a non-foaming composition can be cited, and more preferably, a non-emulsified composition and a non-foaming composition can be cited.

[0075] 2-1-5. Preparation method The liquid composition can be prepared in the form of containing vegetable protein and oil in water, and the preparation method thereof is not particularly limited. For example, the following methods can be cited: a method of mixing water and a substance containing vegetable protein and vegetable oil as a processing material of a vegetable material (and other components as required); a method of mixing water, a substance containing at least vegetable protein as a processing material of a vegetable material, and a substance containing at least vegetable oil as a processing material of a vegetable material (and other components as required); a method of crushing and dispersing a vegetable material containing vegetable protein and vegetable oil in water, and mixing other components as required; a method of crushing and dispersing a vegetable material containing at least vegetable protein in water, and then mixing oil and other components as required, etc.

[0076] 2-2. Enzyme treatment process In the enzyme treatment step, protein deamidase and lipase are allowed to act on the liquid composition. From the viewpoint of further improving the emulsification and / or foaming properties, it is preferred to use a protease in combination with the protein deamidase and lipase.

[0077] 2-2-1. Specified enzymes Details of specific examples of protein deamidase, lipase, and protease are as described in the above-mentioned "1-1-1. Protein deamidase", "1-1-2. Lipase", and "1-1-3. Protease", respectively.

[0078] The ratios of lipase and protease used relative to protein deamidase are as described in the above-mentioned "1-1-2. Lipase" and "1-1-3. Protease", respectively.

[0079] The specific amount of the protein deamidase used is not particularly limited, and as the amount per 1g of vegetable protein, for example, 0.05U or more, or 0.1U or more can be mentioned. From the viewpoint of further improving the emulsification and / or foaming properties, preferably 0.4U or more can be mentioned, more preferably 1U or more, 2U or more, 3U or more, 3.5U or more, or 4U or more can be mentioned, further preferably 4.5U or more, and further preferably 4.8U or more can be mentioned. The upper limit of the range of the amount of the protein deamidase used is not particularly limited, and as the amount per 1g of vegetable protein, for example, 50U or less, 40U or less, 30U or less, 20U or less, or 15U or less can be mentioned, preferably 10U or less, more preferably 7U or less, or 6U or less.

[0080] The specific amount of lipase used is not particularly limited. For example, the amount per 1g of fat is 1U or more. From the viewpoint of further improving the emulsification and / or foaming effects, preferably 5U or more, 10U or more, 14U or more, 30U or more, 40U or more, or 50U or more, more preferably 60U or more, 65U or more, 70U or more, or 73U or more. The upper limit of the range of the amount of lipase used is not particularly limited. For example, the amount per 1g of fat is 500U or less, 400U or less, 350U or less, or 300U or less. From the viewpoint of further improving the emulsification and / or foaming effects, preferably 200U or less, or 170U or less, more preferably 130U or less, or 120U or less, and more preferably 90U or less, 85U or less, or 80U or less.

[0081] In the present invention, when a protease is further combined, the specific amount of the protease used is not particularly limited, and the amount per 1g of vegetable protein is, for example, 1U or more, and from the viewpoint of further improving the emulsification and / or foaming effects, preferably 5U or more, more preferably 10U or more, and even more preferably 13U or more. The upper limit of the range of the amount of the protease used is not particularly limited, and the amount per 1g of vegetable protein is, for example, 150U or less, 100U or less, or 75U or less, and from the viewpoint of further improving the emulsification and / or foaming effects, preferably 60U or less, more preferably 50U or less, even more preferably 40U or less, even more preferably 30U or less, even more preferably 20U or less, or 17U or less.

[0082] 2-2-2. Processing conditions, etc. In the enzyme treatment step, a reaction to improve the emulsification and foaming properties of the liquid composition proceeds through the treatment.

[0083] The order in which the enzymes are allowed to act is not particularly limited, and the enzymes may be allowed to act sequentially in any order, or all the enzymes may be allowed to act simultaneously. It is preferred that all the enzymes be allowed to act simultaneously.

[0084] The reaction conditions (temperature, pH, time, etc.) in the enzyme treatment are appropriately selected according to the temperature and pH characteristics of the enzyme used, the scale of the liquid composition, and the degree of the desired effect of improving emulsification or foaming properties.

[0085] The reaction temperature can be appropriately determined by those skilled in the art according to the temperature characteristics of the enzyme used, and is, for example, 20°C to 60°C, preferably 35°C to 56°C, and more preferably 45°C to 52°C.

[0086] The reaction pH (pH of the liquid composition) can be appropriately determined by those skilled in the art according to the pH characteristics of the enzyme used, and the pH at 25° C. is, for example, 5 to 8 or 5 to 7, preferably 5.5 to 7, more preferably 6 to 6.9, and even more preferably 6.3 to 6.9.

[0087] The treatment time can be appropriately determined by those skilled in the art according to the scale of the liquid composition and the degree of the desired effect of improving emulsification or foaming properties, and may be, for example, 10 minutes to 4 hours, preferably 0.5 hours to 3 hours.

[0088] During the enzyme treatment step, the reaction solution is preferably stirred. The stirring speed can be appropriately determined by a person skilled in the art according to the scale of the liquid composition, and for example, 100 rpm to 300 rpm can be cited, preferably 150 rpm to 250 rpm can be cited, and more preferably 180 rpm to 220 rpm can be cited.

[0089] 2-3. Other processes The method of the present invention may include any other steps in addition to the above-mentioned enzyme treatment step. As a specific example of other steps, as a step performed before the enzyme treatment step, a step of preparing the above-mentioned liquid composition can be cited, and as a step performed after the enzyme treatment step, an emulsification step (in the case of an emulsification improvement method and a foaming improvement method) and a foaming step (in the case of a foaming improvement method) can be cited. The details of the process for preparing the liquid composition are as described in the above-mentioned "2-1-5. Preparation method". As conditions for the emulsification process, as temperature, the above-mentioned reaction temperature can be cited, as stirring conditions, for example, 3000 to 7000 rpm can be cited, preferably 4000 to 6000 rpm can be cited, and more preferably 4500 to 5500 rpm can be cited, and as stirring time, for example, 2 to 20 minutes can be cited, preferably 5 to 15 minutes can be cited, and more preferably 8 to 13 minutes can be cited. As conditions for the foaming process, conditions commonly used for foaming using animal milk can be appropriately adopted.

[0090] 3. Method for producing an emulsion of a liquid composition containing vegetable protein and fat The method for producing an emulsion of a liquid composition containing vegetable protein and fat of the present invention is characterized by comprising an enzyme treatment step of treating the liquid composition containing vegetable protein and fat with protein deamidase and lipase.

[0091] Details of the liquid composition, enzyme treatment step and other steps are as described in the above-mentioned "2. Method for improving the emulsification property of a liquid composition containing vegetable protein and fat, and method for improving the foaming property of a liquid composition containing vegetable protein and fat".

[0092] 4. Emulsion of liquid composition containing vegetable protein and oil The emulsion of the liquid composition containing vegetable protein and fat of the present invention is an emulsified composition obtained by the above-mentioned "3. Method for producing an emulsion of a liquid composition containing vegetable protein and fat", and is a food or beverage. The emulsion of the present invention has excellent emulsification and foaming properties.

[0093] The emulsion of the present invention may be of an oil-in-water type or a water-in-oil type, and preferably, the oil-in-water type is used.

[0094] Specific forms of the food and drink of the emulsion according to the present invention include vegetable protein drinks (vegetable milk, etc.), vegetable creamers (coffee creamer, etc.), vegetable foamed milk, vegetable creamers, and the like.

[0095] Example Hereinafter, the present invention will be specifically described with reference to Examples, but the present invention is not construed as being limited to the following Examples.

[0096] [Materials used] The details of the materials (enzymes, substrates, additives) used in the following test examples are shown below.

[0097] [Table 1] [Enzyme activity determination method] (1) Protein deamidase activity assay 0.1 mL of a sample solution containing protein deamidase was added to 1 mL of 0.2 M phosphate buffer (pH 6.5) containing 30 mM Z-Gln-Gly, and the mixture was allowed to stand at 37°C for 10 minutes, followed by the addition of 1 mL of 0.4 M TCA solution to stop the reaction. As a blank, 1 mL of a 0.4 M TCA solution was added to 1 mL of 0.2 M phosphate buffer (pH 6.5) containing 30 mM Z-Gln-Gly, and 0.1 mL of a sample solution containing protein deamidase was further added, followed by the addition of 1 mL of the solution, and the mixture was allowed to stand at 37°C for 10 minutes.

[0098] The amount of ammonia generated in the reaction solution was measured using Ammonia-test Wako (Fujifilm Wako Pure Chemical Industries, Ltd.). The ammonia concentration in the reaction solution was determined from a calibration curve showing the relationship between ammonia concentration and absorbance (630 nm) prepared using an ammonia standard solution (ammonium chloride).

[0099] Will generate 1 in 1 minute μ The activity of protein deamidase was calculated by the following formula, with the amount of enzyme per mol of ammonia as 1 unit (1U). In the formula, the amount of reaction solution is 2.1, the amount of enzyme solution is 0.1, and Df is the dilution ratio of the enzyme solution. In addition, 17.03 is the molecular weight of ammonia.

[0100] [Number 1] (2) Lipase activity assay A neutral olive oil emulsion as a substrate was prepared by the following method. 200 g of anhydrous sodium carbonate was dissolved in 400 mL of water, and added to about 1.5 L of olive oil that had been heated at 45°C for 20 minutes in advance while gently stirring so as not to emulsify, and then stirred at room temperature for about 15 minutes. After leaving it at room temperature for more than 20 hours, the oil was centrifuged (5°C, 5000 min). -1 , 20 minutes), prepare neutral olive oil. Put 51.4 mL of neutral olive oil and 158 mL of 11 wt% gum arabic and 1.25 wt% calcium chloride dihydrate solution into a homogenizer container and simmer for 16000 ± 500 min. -1 Emulsification was performed for 30 minutes to prepare the product.

[0101] Add 9 mL of water, 2 mL of 0.5 wt% sodium taurocholate test solution, and 24 mL of substrate neutral olive oil emulsion to a flat-bottomed test tube and mix, and place at 37±0.5°C for 10 to 15 minutes. Immerse the pH electrode and the 0.02 mol / L sodium hydroxide solution (for quantitative use) injection tube in the liquid, and then stir continuously. Use 0.02 mol / L sodium hydroxide solution (for quantitative use) to adjust the pH to 7.00 and make the biuret reach "0". Add 5 mL of the sample solution containing lipase to start the reaction. Continue to drop 0.02 mol / L sodium hydroxide solution (for quantitative use) in a manner that maintains pH 7.00. After 10 minutes, add 0.02 mol / L sodium hydroxide solution (for quantitative use) to make the pH reach 9.00. Record the amount of 0.02 mol / sodium hydroxide solution (for quantitative use) used (V10 (mL)). The blank solution was operated in the same manner to adjust the pH to 9.00, and then 5 mL of the sample solution containing lipase was added to adjust the pH to 9.00 again. The amount of 0.02 mol / L sodium hydroxide solution (for quantitative determination) used (V0 (mL)) was recorded.

[0102] Under these conditions, 1 will be generated within 1 minute. μ The amount of enzyme that increases mol of free fatty acid is defined as 1 unit (1U) and is calculated by the following mathematical formula.

[0103] [Number 2] V0: Blank titration value (mL) V10: titration value of reaction solution (mL) f: Factor of 0.02mol / L sodium hydroxide solution (for quantitative use) n: dilution factor per 1g or 1mL of sample (3) Protease activity assay After heating 5 mL of 0.6% (w / v) casein solution (0.7% (w / v) lactic acid, pH 3.0) at 37°C for 10 minutes, add 1 mL of a sample solution containing a protease and immediately shake to mix. After the liquid is placed at 37°C for 10 minutes, add 5 mL of a 0.44 mol / L trichloroacetic acid test solution and shake to mix, place it at 37°C for 30 minutes again, and filter. Remove the initial filtrate 3 mL, weigh the next filtrate 2 mL, add 5 mL of a 0.55 mol / L sodium carbonate test solution and 1 mL of a Folin test solution (1→3), shake and mix thoroughly, and place it at 37°C for 30 minutes. For this liquid (enzyme reaction solution), use water as a control and measure the absorbance AT at a wavelength of 660 nm. In addition, 1 mL of the sample solution containing protease was weighed, 5 mL of 0.44 mol / L trichloroacetic acid test solution was added and shaken, and then 5 mL of 0.6% (w / v) casein solution (0.7% (w / v) lactic acid, pH 3.0) was added, and immediately shaken and mixed, and then placed at 37°C for 30 minutes. In addition, the liquid (blank) obtained by the same operation as the above enzyme reaction solution was measured for absorbance AB. μ The amount of enzyme added to produce the color-developing substance in g of Folin test solution is taken as 1 unit (1U).

[0104] Weigh 1mL, 2mL, 3mL and 4mL of 1mg / mL tyrosine standard stock solution (0.2 mol / L hydrochloric acid), and add 0.2mol / L hydrochloric acid test solution to make 100mL respectively. Weigh 2mL of each liquid, add 5mL of 0.55 mol / L sodium carbonate test solution and 1mL of Folin test solution (1→3), immediately shake to mix, and place at 37℃ for 30 minutes. For these liquids, measure 2mL of 0.2 mol / L hydrochloric acid test solution, and use the liquid obtained by the same operation as above as a control to measure the absorbance A1, A2, A3 and A4 at a wavelength of 660nm. Take the absorbance A1, A2, A3 and A4 on the vertical axis, and take the amount of tyrosine in 2mL of each liquid on the horizontal axis ( μ g), make a standard curve, and calculate the amount of tyrosine corresponding to the absorbance difference of 1 ( μ g).

[0105] [Number 3] AT: absorbance of enzyme reaction solution AB: absorbance of blank F: The amount of tyrosine when the absorbance difference is 1, obtained from the tyrosine standard curve ( μ g) 11 / 2: Conversion factor for the total liquid volume after the reaction stops 1 / 10: Conversion factor for every 1 minute of reaction time M: The amount of sample in 1 mL of sample solution (g or mL) [Test Example 1] Rapeseed oil was added to the protein solution prepared by suspending the pea protein raw material in 0.1M sodium phosphate buffer (pH 6.8), and stirred at 50°C for 10 minutes to prepare the liquid composition of the composition shown in Tables 2 to 5, and then the enzyme in the amount shown in Tables 2 to 5 was added, and the reaction was carried out at 50°C and 200 rpm for 2 hours. Then, the enzyme was treated at 85°C for 15 minutes to inactivate the enzyme. The reaction solution was emulsified at 5000 rpm for 10 minutes using a homogenizer (SILERSON, L5M-A), and then cooled to room temperature (25°C) to prepare an oil-in-water emulsified composition (pH at 25°C: 6.8).

[0106] 35 mL of the obtained emulsified composition was taken into three tubes of 50 mL capacity, respectively, and analytical samples 1, 2, and 3 were prepared as follows.

[0107] Analysis sample 1: an emulsified composition immediately after preparation (day 0, 25° C.).

[0108] Analysis sample 2: The emulsified composition was left to stand in a refrigerator set at 4° C. for 1 week.

[0109] Analysis sample 3: The emulsified composition was centrifuged at 3000 rpm for 25 minutes (25° C.) Centrifugation at 3000 rpm for 25 minutes is an accelerated condition equivalent to standing for 1 year (Procedia Chemistry, Volume 16, 2015, Pages 171-176).

[0110] 1 mL was sampled from each of samples 1 to 3 at a position 1 cm below the upper end of the emulsified phase, and diluted 100 times with purified water. For the 100-fold diluted sample obtained, the absorbance at 280 nm and the absorbance at 600 nm were measured. After confirming the solubilization of the protein by the absorbance at 280 nm, the degree of emulsification was evaluated by the absorbance at 600 nm. The greater the absorbance at 600 nm, the greater the improvement in emulsification. The results are shown in Tables 2 to 5.

[0111] [Table 2] The magnification of absorbance in the examples indicates a relative value when the absorbance in Comparative Example 1 is set to 1.

[0112] [Table 3] The magnification of the absorbance in the examples indicates a relative value when the absorbance in Comparative Example 2 is set to 1.

[0113] [Table 4] The magnification of the absorbance in Examples 10 and 11 indicates a relative value when the absorbance in Comparative Examples 3 and 4 is set to 1, respectively.

[0114] [Table 5] The magnification of the absorbance in Examples 12, 13, and 14 indicates a relative value when the absorbance in Comparative Examples 5, 6, and 7 is set to 1, respectively.

[0115] As can be seen from Tables 2 to 5, the emulsification property is improved by treating the liquid composition containing vegetable protein and oil with protein deamidase and lipase.

[0116] [Test Example 2] RO water, pea protein raw material, almond fat and cane sugar, and sunflower oil and / or lecithin added as needed were mixed, and stirred at 5000 rpm for 10 minutes using a homogenizer (SILERSON, L5M-A) to prepare a liquid composition of the composition shown in Tables 6 and 7. The amount of total protein and total lipid in the liquid composition is shown in Tables 6 and 7. The amount of enzyme shown in Tables 6 and 7 was added to the liquid composition, and stirred at 50°C for 1 hour (200 rpm). Then, emulsification was performed at 5000 rpm for 10 minutes, the enzyme was inactivated by heat treatment (85°C, 15 minutes), and further cooled to room temperature (25°C) to prepare an oil-in-water emulsified composition (almond milk) (pH at 25°C: 6.5).

[0117] In a 50 mL tube, the prepared almond milk was divided into 100mL (sample for evaluation of solubility and emulsification) and 100mL (sample for evaluation of foaming).

[0118] For the samples for evaluating solubility and emulsification, the same analytical samples as analytical samples 1 and 3 of Test Example 1 were prepared, and 100-fold diluted samples were prepared in the same manner as in Test Example 1, and the absorbance at 280 nm and the absorbance at 600 nm were measured. The results are shown in Tables 6 and 7.

[0119] Separately, the sample for foamability evaluation was foamed using a milk frother (HadinEEon, model: MMF-9401) to prepare foamed milk. The obtained foamed milk was transferred to a measuring cylinder, and the volume of the foam layer was measured to evaluate the foamability. The larger the volume, the better the foamability. The results are shown in Tables 6 and 7.

[0120] [Table 6] The magnification display of the absorbance and foaming property in Reference Example 1 and Examples 15 and 16 indicates a relative value when the absorbance and foaming property in Comparative Example 8 are set to 1.

[0121] [Table 7] The magnification display of the absorbance and foaming property in Reference Example 2 and Example 17 shows the relative value when the absorbance and foaming property in Comparative Example 9 are set to 1.

[0122] As shown in Tables 6 and 7, the emulsification property of the liquid composition containing vegetable protein and oil is improved by treating it with protein deamidase and lipase. The degree is improved compared with the case where lecithin, which is a general-purpose emulsifier, is used. In particular, when protease is used together with protein deamidase and lipase, the emulsification property is further improved.

[0123] In addition, it can be seen from Tables 6 and 7 that the foaming property of the liquid composition containing vegetable protein and fat is also improved by treating it with protein deamidase and lipase. In particular, when protease is used together with protein deamidase and lipase, the foaming property is further improved, and the degree is improved compared with the case of using lecithin, which is a general-purpose emulsifier.

Claims

1. An emulsification enhancer for a liquid composition, characterized in that: The liquid composition contains vegetable protein and oil, and the emulsification enhancer contains protein deamidase and lipase.

2. The emulsification enhancer according to claim 1, wherein The amount of the lipase is 1 U or more relative to 1 U of the protein deamidase.

3. The emulsification enhancer according to claim 1, wherein The emulsification enhancer further comprises a protease.

4. A foaming enhancer for a liquid composition, characterized in that The liquid composition comprises vegetable protein and oil, and the foaming enhancer comprises protein deamidase and lipase.

5. The foaming property enhancing agent according to claim 4, wherein The foaming enhancer further comprises a protease.

6. The foaming property enhancing agent according to claim 5, wherein The amount of the protease is 1 U or more relative to 1 U of the protein deamidase.

7. A method for improving the emulsification of a liquid composition, characterized in that: The liquid composition contains vegetable protein and oil, and the method includes an enzyme treatment step of allowing protein deamidase and lipase to act on the liquid composition.

8. The method according to claim 7, wherein: 0.05 U or more of the protein deamidase is used per 1 g of the vegetable protein.

9. The method according to claim 7, wherein: At least 1 U of the lipase is used per 1 g of the oil or fat.

10. A method for improving the foamability of a liquid composition, characterized in that: The liquid composition contains vegetable protein and oil, and the method includes an enzyme treatment step of allowing protein deamidase and lipase to act on the liquid composition.

11. A method for producing an emulsion of a liquid composition, characterized in that: The liquid composition comprises vegetable protein and oil, and the method comprises an enzyme treatment step of treating the liquid composition with protein deamidase and lipase.

12. The manufacturing method according to claim 11, wherein: In the enzyme treatment step, protease is further used.

13. A food or beverage, characterized in that: An emulsion of a liquid composition containing vegetable protein and oil obtained by the production method of claim 11.

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