Thickening agent for composition containing vegetable protein

Abstract

The purpose of the present invention is to provide a novel processing technique capable of thickening a composition containing a vegetable protein. In the present invention, when a plant protein-containing composition comprising chia seed protein and / or hemp seed protein is treated with a protein deamidase, the plant protein-containing composition can be thickened.

Description

Technical Field

[0001] The present invention relates to a thickener for a composition containing vegetable protein. More specifically, the present invention relates to a processing technology for thickening a composition containing vegetable protein, wherein the composition containing vegetable protein comprises chia seed protein and / or hemp seed protein. Background Art

[0002] Nutritious plant milk is gaining popularity as an alternative to animal milk, due to various background factors, including the recent health craze, strategies to combat allergies, and religious reasons.

[0003] Animal milk has the characteristic of being able to be transformed into various forms by undergoing specific processing, and a large number of dairy products have been created so far, which have been widely accepted by consumers. On the other hand, plant milk has a fundamentally different composition from animal milk, so when manufacturing processed products such as alternative milk products, completely different methods are generally required compared to when animal milk is used as the raw material.

[0004] One of the changes in form required in the processing of vegetable milk is thickening. For such thickening, it is known to use, for example, processed starches such as acid-treated starch derived from peas or potatoes, hydroxypropylated starch, and thickeners such as locust bean gum and guar gum (Patent Document 1).

[0005] Prior art literature Patent Literature Patent Document 1: International Publication No. 2018 / 173610 Summary of the invention

[0006] Technical problem to be solved by the invention In order to further enrich the diversity of animal milk substitute foods and further increase the market share in the food and beverage market, a new thickening processing technology is desired that can be widely used in various cooking methods.

[0007] Therefore, an object of the present invention is to provide a new processing technology capable of thickening a composition containing vegetable protein.

[0008] Technical solutions for solving technical problems The present inventors have discovered that by treating a composition containing chia seed protein and a composition containing hemp seed protein with a protein deamidase, the composition can be thickened. Considering that the thickening effect of protein deamidase is not known and that a thickening effect cannot be confirmed in a large number of other compositions containing plant proteins, it is extremely difficult to predict that a composition containing chia seed protein and a composition containing hemp seed protein would have a unique thickening effect. The present invention was completed by further repeated studies based on this understanding.

[0009] That is, the present invention provides the following inventions.

[0010] Item 1. A thickener for thickening a composition containing vegetable protein, the thickener comprising a protein deamidase, wherein the composition containing vegetable protein comprises chia seed protein and / or hemp seed protein.

[0011] Item 2. The thickener according to Item 1, wherein the total amount of the chia seed protein and the hemp seed protein in the plant protein-containing composition is 4% by weight or more.

[0012] Item 3. A method for producing a thickened vegetable protein-containing composition, the method comprising allowing a protein deamidase to act on the vegetable protein-containing composition, wherein the vegetable protein-containing composition comprises chia seed protein and / or hemp seed protein.

[0013] Effects of the Invention According to the present invention, a new processing technology capable of thickening a composition containing a vegetable protein is provided. DETAILED DESCRIPTION

[0014] 1. Thickener The thickener of the present invention is characterized in that it contains a protein deamidase and is used for thickening a composition containing vegetable protein, wherein the composition containing vegetable protein contains chia seed protein and / or hemp seed protein. The thickener of the present invention is described in detail below.

[0015] 1-1. Application The thickener of the present invention is used for thickening a composition containing vegetable protein, wherein the composition containing vegetable protein comprises chia seed protein and / or hemp seed protein.

[0016] The composition containing vegetable protein to which the thickener of the present invention is applied is not particularly limited as long as it is a composition containing vegetable protein comprising chia seed protein and / or hemp seed protein and is a composition having fluidity. As specific properties of the composition containing vegetable protein, liquid, slurry and paste can be cited. The thickener of the present invention has an excellent thickening effect, so as a preferred property of the composition containing vegetable protein, liquid can be cited.

[0017] The vegetable protein-containing composition in the form of a liquid (hereinafter also referred to as a "liquid composition containing vegetable protein") is not particularly limited as long as it is a liquid containing chia seed protein and / or hemp seed protein in water. Specific examples of liquid compositions containing vegetable protein include: (i) a liquid obtained by dispersing a dry powder of a food material containing chia seed protein and / or hemp seed protein (specifically, at least one of plant organs selected from chia seeds and hemp seeds, and materials in which the content of the protein is increased by removing at least a portion of components other than protein from the plant organs) in water; (ii) a liquid obtained by crushing and dispersing chia seeds and / or hemp seeds in water, and removing insoluble matter derived from the skin of these seeds by any means such as centrifugation, filtration, filter bag, sieve, etc. as needed; (iii) a liquid obtained by removing components other than chia seed protein and / or hemp seed protein from the liquid of (i) or (ii) to increase the content of the protein; (iv) a liquid obtained by mixing a dry powder prepared from any of the liquids (i) to (iii) with water, etc. Preferred examples of compositions containing vegetable protein include vegetable milk.

[0018] In addition, the chia seed protein and / or hemp seed protein contained in the liquids of (i) to (iv) above may be in any of the following forms: protein contained in chia seeds and / or hemp seeds (natural protein); protein partially chemically decomposed by acid, alkali, etc. treatment; partially enzymatically hydrolyzed protein obtained by protease, etc.; chemically modified protein obtained by various reagents, etc. Among these proteins, chia seed protein is preferred from the viewpoint of further improving the thickening effect.

[0019] It should be noted that, hereinafter, when the “content of plant protein material” in a composition containing plant protein is recorded, it refers to the proportion of the dry weight of the components constituting the above-mentioned plant organs contained in the composition containing plant protein. For example, in the case where the composition containing plant protein is a liquid composed only of components derived from plant organs and water, such as the liquids (i) to (iv) above, the “content of plant protein material” refers to the proportion of the dry weight of the liquid. In addition, for example, in the case where the composition containing plant protein is a liquid containing the liquids (i) to (iv) above and additives, the “content of plant protein material” refers to the proportion of the dry weight of the portion after the additives are removed from the liquid.

[0020] The content of chia seed protein and / or hemp seed protein in the composition containing vegetable protein to which the thickener of the present invention is applied is not particularly limited; based on the total content of chia seed protein and hemp seed protein, for example, it can be 4% by weight or more, preferably 5.5% by weight or more, more preferably 6% by weight or more, further preferably 7% by weight or more, and further preferably 8% by weight or more.

[0021] The upper limit of the chia seed protein and / or hemp seed protein content in the composition containing plant protein is not particularly limited, and can be, for example, 20 wt % or less, preferably 17 wt % or less, more preferably 13 wt % or less, further preferably 11 wt % or less, or 10 wt % or less, based on the total amount of chia seed protein and hemp seed protein.

[0022] Regarding the content of chia seed protein material and / or hemp seed protein material (chia seed protein material and hemp seed protein material are collectively referred to as "plant protein material") in the composition containing plant protein, based on the total content of the plant protein material, for example, it can be 4% by weight or more, preferably 5.5% by weight or more, more preferably 6% by weight or more, further preferably 7% by weight or more, further preferably 8% by weight or more, and further preferably 9% by weight or more; as its upper limit, for example, it can be 20% by weight or less, preferably 17% by weight or less, more preferably 13% by weight or less, and further preferably 11% by weight or less.

[0023] 1-2. Protein deamidase The thickener of the present invention contains protein deamidase as an active ingredient. The thickener of the present invention can thicken the obtained vegetable protein-containing composition based on the action of the protein deamidase on the vegetable protein-containing composition containing chia seed protein and / or hemp seed protein.

[0024] As the protein deamidase used in the present invention, there is no particular limitation on its type and source, etc., as long as it is an enzyme that can show the action of decomposing the amide-containing side chain of the protein without the cleavage of the peptide bond and the cross-linking of the protein. In addition, as long as the above-mentioned action is the main activity, it may further have the action of decomposing the amide-containing side chain of the protein with the cleavage of the peptide bond and the cross-linking of the protein.

[0025] Examples of protein deamidase include enzymes that deamidate glutamine residues in proteins and convert them to glutamate (e.g., protein glutaminase), enzymes that deamidate asparagine residues in proteins and convert them to aspartic acid (e.g., protein asparaginase), and enzymes that deiminate the guanidine group of the arginine residue at the C-terminus of proteins (e.g., protein arginine deiminase). Among them, protein glutaminase is preferred from the viewpoint of further improving the thickening effect.

[0026] 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.

[0027] Among these protein deaminases, from the viewpoint of further improving the thickening effect, preferably, a protein deamidase derived from the genus Chryseobacterium is cited, more preferably, a protein glutaminase derived from the genus Chryseobacterium is cited, further preferably, a protein glutaminase derived from the species Chryseobacterium proteolyticum is cited, and further preferably, a protein glutaminase derived from Chryseobacterium proteolyticum is cited.

[0028] 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. as needed, and then 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, lysozyme 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 separated and / or purified enzyme 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.

[0029] The amount of the protein deamidase contained in the thickener of the present invention is not particularly limited as long as the effect of the protein deamidase can be effectively obtained when the thickener of the present invention is used, and examples thereof include 0.1 U / g to 5000 U / g.

[0030] Regarding the activity of protein deamidase, benzyloxycarbonyl-L-glutaminoglycine (Z-Gln-Gly) was used as a substrate and 1 μ The amount of enzyme per mol of ammonia is taken as 1 unit (1U).

[0031] 1-3. Other ingredients The thickener of the present invention may or may not contain other components in addition to the above-mentioned protein deamidase to the extent that the effects of the present invention are not impaired. Examples of other components include enzymes other than protein deamidase, additives, and the like.

[0032] As other enzymes, one or more enzymes can be appropriately selected based on the type and composition of the vegetable protein-containing composition to which the thickener of the present invention is applied, and in accordance with a desired reaction other than the reaction using the protein deamidase.

[0033] As additives, it is possible to appropriately select according to the desired formulation form. The formulation form of the thickener of the present invention is not particularly limited either, and can be any of a solid preparation and a liquid preparation. Therefore, as additives, for example, excipients, buffers, suspending agents, stabilizers, preservatives, preservatives, water, pH adjusting agents, etc. can be cited. These additives can include one alone, or a combination of multiple ones.

[0034] 2. Method for producing a thickened vegetable protein-containing composition The present invention also provides a method for producing a thickened vegetable protein-containing composition, which comprises a step of allowing a protein deamidase to act on the vegetable protein-containing composition, wherein the vegetable protein-containing composition comprises chia seed protein and / or hemp seed protein.

[0035] 2-1. Step of allowing protein deamidase to act The type of the composition containing vegetable protein and the type of the protein deamidase in the step of allowing the protein deamidase to act are as shown in the above-mentioned column "1. Thickener".

[0036] There is no particular limitation on the amount of protein deamidase used. For example, the amount of protein deamidase used is 0.25U or more, preferably 0.5U or more, per 1g of the total amount of chia seed protein and hemp seed protein contained in the composition containing vegetable protein. From the viewpoint of further improving the thickening effect, the amount of protein deamidase used is preferably 1U or more, more preferably 2U or more, further preferably 3U or more, further preferably 4U or more, 5U or more, or 5.5U or more, per 1g of the total amount of chia seed protein and hemp seed protein contained in the composition containing vegetable protein. The upper limit of the amount of protein deamidase used is not particularly limited, and for example, 500U or less, 250U or less, 100U or less, 75U or less, 50U or less, 25U or less, 10U or less, or 7.5U or less.

[0037] In addition, regarding the amount of protein deamidase used, as the amount of protein deamidase used per 1 g (dry weight) of the total amount of vegetable protein material contained in the composition containing vegetable protein, for example, 0.25 U or more, preferably 0.5 U or more. From the viewpoint of further improving the thickening effect, as the amount of protein deamidase used per 1 g (dry weight) of the total amount of vegetable protein material contained in the composition containing vegetable protein, preferably 1 U or more, more preferably 2 U or more, further preferably 3 U or more, and further preferably 4 U or more or 4.5 U or more. As the upper limit of the range of the amount of protein deamidase used per 1 g (dry weight) of the total amount of vegetable protein material contained in the composition containing vegetable protein, there is no particular limitation, and for example, 500 U or less, 250 U or less, 100 U or less, 75 U or less, 50 U or less, 25 U or less, 10 U or less, or 7.5 U or less can be mentioned.

[0038] In the step of allowing the protein deamidase to act, the composition containing vegetable protein is mixed with the protein deamidase, and the protein deamidase is allowed to act on the protein in the composition containing vegetable protein, thereby performing a viscosity-increasing reaction. Regarding the reaction conditions, those skilled in the art can appropriately set the reaction temperature and pH for the protein deamidase to act on the protein in the composition containing vegetable protein, and the reaction time for allowing the reaction to proceed to a sufficient degree of thickening, etc., by taking into account the temperature and pH characteristics of the protein deamidase used, through preliminary tests, etc.

[0039] The reaction temperature can be determined according to the optimum temperature of the protein deamidase used, and examples thereof include 10° C. to 80° C., preferably 20° C. to 70° C., more preferably 40° C. to 60° C., and further preferably 45° C. to 55° C. The pH can be determined according to the optimum pH of the protein deamidase used, and examples thereof include 5 to 10, preferably 5 to 8, and more preferably 6 to 8 at 25° C. The reaction time can be determined according to the reaction scale, and examples thereof include 1 hour to 48 hours, preferably 1 hour to 24 hours, and more preferably 8 hours to 20 hours or 10 hours to 18 hours.

[0040] 2-2. Other processes The production method of the present invention may include other steps in addition to the above-mentioned step of allowing the protein deamidase to act.

[0041] Specific examples of other steps include a protein deamidase inactivation step, a cooling step, and a filtration step.

[0042] 2-3. Thickened vegetable protein-containing composition The thickened vegetable protein-containing composition obtained by the production method of the present invention has a higher viscosity than the vegetable protein-containing composition not treated with the thickener of the present invention. The thickened vegetable protein-containing composition can be used directly or by appropriate cooking, for example, as a vegetable milk substitute, cream substitute, yogurt substitute, cheese substitute, ice cream substitute, etc.

[0043] 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.

[0044] [Enzymes used] As the protein deamidase, PG, which is a protein glutaminase derived from Chryseobacterium proteolyticum (manufactured by Amano Enzyme Co., Ltd.), was used.

[0045] [Enzyme activity determination method] 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 placed 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 control, 1 mL of 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 mixture, and the mixture was placed at 37°C for 10 minutes.

[0046] The obtained solution was measured for the amount of ammonia generated in the reaction solution using Ammonia test Wako (Fujifilm Wako Pure Chemical Industries, Ltd.). A standard curve showing the relationship between ammonia concentration and absorbance (630 nm) was prepared using an ammonia standard solution (ammonium chloride), and the ammonia concentration in the reaction solution was determined from the standard curve.

[0047] Will be able to 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.

[0048] [Number 1] Protein deamidase activity (U / mL) = ammonia concentration in reaction solution (mg / L) × (1 / 17.03) × (reaction solution volume / enzyme solution volume) × (1 / 10) × Df [Plant protein materials] Use the plant-based protein materials (all powders) shown in the table below.

[0049] [Table 1]

[0050] [Test example] (1) Test operation The various vegetable protein materials in Table 1 were mixed with a phosphate buffer solution at pH 7.0 (25° C.) so that the vegetable protein material was 10% by weight, to prepare a liquid composition containing vegetable protein. 5 U of PG was added per 1 g of the vegetable protein material to the liquid composition containing vegetable protein, and the mixture was incubated at 50° C. overnight to obtain a treated composition containing vegetable protein (“with” PG). In addition, the same operation was performed except that PG was not added to obtain a treated composition containing vegetable protein (“without” PG).

[0051] (2) Viscosity evaluation 30 mL of the PG-treated vegetable protein composition obtained in (1) above was added to a 50 mL tube, and the degree of viscosity that can be grasped based on the movement speed of the liquid during inversion mixing at room temperature (25°C) was compared with a vegetable protein composition that was not treated with PG (no PG). It should be noted that regarding inversion mixing, the operation of inverting 180° and then resetting by -180° is considered as one cycle, and the cycle is performed at a speed of about 2 seconds to complete one cycle. The slower the movement speed of the vegetable protein-containing composition relative to the inversion speed, the higher the viscosity can be evaluated. The degree of thickening of the vegetable protein-containing composition after PG treatment based on this comparison is classified based on the following criteria. The results are shown in Table 2.

[0052] XX: The viscosity is significantly reduced compared to the case without PG.

[0053] ×: The viscosity is lower than that in the case without PG.

[0054] -: No change compared to the case without PG

[0055] ○: The viscosity increased compared to the case without PG.

[0056] ◎: The viscosity is significantly increased compared to the case without PG.

[0057] [Table 2]

[0058] As shown in Table 2, thickening was confirmed by subjecting the liquid composition containing chia seed protein and the liquid composition containing hemp seed protein to PG treatment (Examples 1 and 2), and the effect was particularly significant in the case of the liquid composition containing chia seed protein (Example 1). It should be noted that even with PG treatment, thickening was not confirmed for various other plant proteins represented by almond protein and walnut protein, which are seed proteins like chia seed protein and hemp seed protein (Comparative Examples 1 to 15), and even a decrease in viscosity was confirmed in some of them. Therefore, it is judged that the thickening effect confirmed in Examples 1 and 2 is an effect unique to the liquid composition containing chia seed protein and the liquid composition containing hemp seed protein.

Claims

1. A thickener, characterized in that The thickener is used for thickening a composition containing vegetable protein, the thickener comprises protein deamidase, and the composition containing vegetable protein comprises chia seed protein and / or hemp seed protein.

2. The thickener according to claim 1, wherein The total content of the chia seed protein and the hemp seed protein in the composition containing plant protein is 4% by weight or more.

3. A method for producing a thickened vegetable protein-containing composition, characterized in that: The production method comprises a step of allowing a protein deamidase to act on a composition containing plant proteins, wherein the composition containing plant proteins comprises chia seed protein and / or hemp seed protein.

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