JP2015186477A - PLANT-DERIVED β-GLUCAN CONTAINING SYRUP - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a plant-derived β-glucan containing syrup that has a sufficiently low viscosity and is easy to handle.SOLUTION: A plant-derived β-glucan containing syrup comprises plant-derived β-glucan with a weight average molecular weight of 2,500-40,000, wherein the content of the plant-derived β-glucan is 2 mass%-8 mass% relative to the total soluble solid content.

Description

  The present invention relates to a syrup containing β-glucan derived from a plant, and a food or beverage containing the syrup.

  β-glucan is a linear polymer in which glucose is a constituent unit and these are β-1,3 bonds and β-1,4 bonds. And this β-glucan has been reported to have physiological functions such as normalization of blood cholesterol, suppression of an increase in blood glucose level after meals, and a sustained action of satiety.

  In particular, the following mechanism is known regarding the normalization of blood cholesterol. Cholesterol is absorbed into the small intestine by associating with bile acid micelles and incorporating cholesterol into bile acid micelles. β-glucan inhibits the formation of these micelles or destroys the formed micelles, thereby suppressing the absorption of cholesterol into the small intestine.

  However, such inhibition ability has been confirmed only with a very large β-glucan having a weight average molecular weight derived from oats exceeding 300,000 (Non-patent Document 1), and a relatively small weight average. No such effect has been reported with molecular weight β-glucan.

  By the way, it is known that β-glucan is contained in a cell wall such as barley. Malt syrup is a food material produced by saccharification of malt germinated from barley, but β-glucan is decomposed in the process of germination or saccharification, and is hardly contained in the syrup. Patent Document 1 discloses a syrup containing barley-derived β-glucan having a weight average molecular weight of 50,000 to 500,000, and is used in foods such as bread and yogurt, and beverages such as soft drinks and sake. It is described.

  However, for example, when considering using syrup as a raw material for foods and beverages, the syrup described in Patent Document 1 contains a very high molecular weight β-glucan having a weight average molecular weight of 50,000 to 500,000. There is a problem that the viscosity is too high and handling is difficult. Patent Document 1 also describes that a syrup having a sufficiently low viscosity was obtained, but this was achieved simply by reducing the Brix of the obtained syrup to less than 10%. However, because of its low Brix, it has a low β-glucan concentration, and it has a high risk of microbial growth during storage of syrup, and as a syrup for foods and beverages, it has a problem in its handling. Met.

Japanese Patent No. 5186311

Hyun Jung Kim et al. , Journal of Agricultural and Food Chemistry (2010) vol. 58, pp. 628-634

  An object of the present invention is to provide a plant-derived β-glucan-containing syrup having a sufficiently low viscosity and easy to handle. Another object of the present invention is to provide a plant-derived β-glucan-containing syrup that has the ability to inhibit micellization of cholesterol and is excellent in functionality. Furthermore, it is an object to provide a new food or beverage to which the plant-derived β-glucan-containing syrup is added.

In order to solve the above problems, the inventors of the present invention manufactured a plant-derived β-glucan-containing syrup containing a plant-derived β-glucan having a weight average molecular weight of 2,500 to 40,000, and derived from such a plant. It has been found that the β-glucan-containing syrup has a sufficiently low viscosity and is easy to handle. Therefore, the first aspect of the present invention is
(1) A plant-derived β-glucan containing a plant-derived β-glucan having a weight average molecular weight of 2,500 to 40,000 in a content of 2% by mass to 8% by mass with respect to the entire soluble solid content. Containing syrup.

A preferred embodiment of the present invention is:
(2) The plant-derived β-glucan-containing syrup according to (1) above, wherein the plant-derived β-glucan contained has an ability to inhibit cholesterol micellization.

A preferred embodiment of the present invention is:
(3) The plant-derived β-glucan-containing syrup according to (1) or (2), wherein the viscosity is 20,000 cP or less.

A preferred embodiment of the present invention is:
(4) The plant-derived β-glucan-containing syrup according to any one of (1) to (3) above, wherein Brix is 30% to 80%.

A preferred embodiment of the present invention is:
(5) The plant-derived β-glucan-containing syrup according to any one of (1) to (4) above, wherein the pH is 4.0 to 8.0.

A preferred embodiment of the present invention is:
(6) The plant-derived β-glucan-containing syrup according to any one of (1) to (5) above, wherein the plant-derived β-glucan is a gramineous plant-derived β-glucan.

A preferred embodiment of the present invention is:
(7) A plant-derived β-glucan-containing syrup according to any one of (1) to (6) above, which is obtained by a production method including a reaction step using glucoamylase.

Moreover, it was found that the plant-derived β-glucan-containing syrups (1) to (7) have low viscosity and are easy to handle, and are excellent as raw materials for foods or beverages. Further, it was confirmed that even when the weight average molecular weight is 40,000 or less, the formation of micelles in which cholesterol is dissolved can be inhibited. Accordingly, another aspect of the present invention is:
(8) A food or beverage comprising the plant-derived β-glucan-containing syrup of any one of (1) to (7) above.

  According to the present invention, a plant-derived β-glucan-containing syrup having a sufficiently low viscosity and easy to handle can be provided. Moreover, although it is easy to handle, a plant-derived β-glucan-containing syrup having an ability to inhibit cholesterol micelle formation and excellent functionality can be provided. Furthermore, a new food or beverage to which the plant-derived β-glucan-containing syrup is added can be provided.

FIG. 1 is a diagram showing the results of extracting β-glucan from the plant-derived β-glucan-containing syrup obtained in Example 1 and measuring the molecular weight distribution by gel filtration chromatography. FIG. 2 is a diagram showing the results of extracting β-glucan from the plant-derived β-glucan-containing syrup obtained in Comparative Example 1 and measuring the molecular weight distribution by gel filtration chromatography. FIG. 3 is a diagram showing the results of measuring the relationship between each Brix and viscosity in the plant-derived β-glucan-containing syrup of Example 1. FIG. 4 is a diagram showing the evaluation test results of the ability to suppress the increase in blood glucose level of each syrup taken by subject A. FIG. 5 is a diagram showing the evaluation test results of the ability to suppress the increase in blood glucose level of each syrup taken by subject B.

  In the present invention, a plant-derived β-glucan-containing syrup containing a plant-derived β-glucan having a weight average molecular weight of 2,500 to 40,000 at a content of 2% by mass to 8% by mass with respect to the entire soluble solid content is produced. doing. Such a plant-derived β-glucan-containing syrup is not suggested in the prior art document. Specifically, the present invention can be implemented as follows.

<Plant-derived β-glucan>
As an example of a syrup raw material according to the present invention, a plant containing β-glucan is used. The form of the “plant” containing such β-glucan may be any of leaves, petals, stems, xylem, roots, epidermis, fiber cells, branches, fruits, seeds and the like.

  Preferred examples of the “plant” used as a raw material include rice plants such as rice, wheat, corn, sorghum, barnyard millet, millet, millet, barley, oats, and rye. Preferably, barley and oats are mentioned, More preferably, barley is mentioned. In addition, as a raw material, it selects according to a use and the objective from these plants, and may be used independently, and may be used in combination as appropriate.

  In addition, when using barley as a raw material, barley of arbitrary varieties, such as Nijo barley, Shijo barley, Rojo barley, and bare barley, can be used. Moreover, as the form, arbitrary structures | tissues or fractions of barley seeds, such as whole grain, wheat grains, and straw, can be used. In barley seeds, since the endosperm contains a large amount of β-glucan, it is more preferable to use whole grains including the endosperm and barley grains, and it is particularly preferable to use the barley grains.

<Method for producing plant-derived β-glucan-containing syrup>
Hereinafter, the production method of a plant-derived β-glucan-containing syrup according to the present invention will be described in the case of using barley as a raw material, but the raw material to be used is not limited to barley, and naturally the above plant can be used in addition to barley. It is. This production method is also an example of a method for producing a plant-derived β-glucan-containing syrup, and other production methods can be used as a matter of course.

1. Preparation process First, when manufacturing the syrup which concerns on this invention, the ground barley seeds are mixed with a pure water, stirring suitably. Thereafter, the solution in which the pulverized barley seeds are mixed is stirred in a thermostatic bath at 30 ° C. to 60 ° C. to completely disperse the barley pulverized substances in pure water.

  The concentration of the barley seed pulverized product at this time is preferably 5% to 50%, more preferably 10% to 30% on a mass basis (W / W) based on the total amount of the solution. In addition, when this density | concentration is higher than 50%, sufficient liquefaction reaction cannot be performed in a liquefaction process, undegraded starch remains, and causes a fall of commercial value. Moreover, the solution in which the undegraded starch remains causes the viscosity to increase, and makes it difficult to separate the solid and the liquid in the filtration process and the filtering process.

  Further, the pH of the solution is preferably adjusted to 3.0 to 8.0, more preferably pH 4.0 to 7.0, and still more preferably pH 5.0 to 6.0 using an appropriate pH adjuster. Is desirable. When the pH is 8 or less, coloring due to the reaction is suppressed, and the flavor of the resulting syrup is improved. When the pH is 3 or more, the enzyme activity is high and the reaction can be carried out sufficiently. Any pH adjusting agent may be used as long as it is suitable for food or beverage. For example, the pH adjuster used to lower the pH is preferably hydrochloric acid, phosphoric acid, citric acid, tartaric acid, fumaric acid, adipic acid, acetic acid, succinic acid, sorbic acid, erythorbic acid, potassium dihydrogen phosphate Sodium dihydrogen phosphate or a combination thereof, more preferably citric acid or malic acid, and still more preferably malic acid. Further, as a pH adjuster used for raising the pH, preferably sodium carbonate, potassium carbonate, sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, calcium oxide, magnesium oxide, dipotassium hydrogen phosphate, Disodium hydrogen phosphate or a combination thereof is more preferably calcium hydroxide or sodium hydroxide, and still more preferably calcium hydroxide.

  Thereafter, the proteolytic enzyme is added to the solution in which the barley pulverized product is dispersed, and the temperature of the solution is raised to the reaction temperature of the added proteolytic enzyme, and the reaction is performed for a predetermined reaction time. It is important to carry out the proteolytic reaction before the liquefaction reaction. Since a protein exists around β-glucan, an enzyme cannot approach β-glucan unless a proteolytic reaction is performed. For this reason, finally, only a syrup that does not contain sufficient β-glucan can be obtained.

  As a proteolytic enzyme to be added, it is sufficient if it has an EC number of Group 3.4, and preferably a protease (EC 3.4.22.2) can be used. Such a proteolytic enzyme may be a commercially available one, and particularly preferably, trade name “Sumiteam P” (manufactured by Shin Nippon Chemical Industry Co., Ltd.).

  The amount of proteolytic enzyme added is preferably 0.001% to 2.0%, more preferably 0.01% to 1.0%, still more preferably 0.03% on a mass basis (W / W) relative to the raw material. ~ 0.1%. If it is 2.0% or less, unpleasant flavor can be suppressed, and if it is 0.001% or more, the proteolytic reaction proceeds sufficiently.

  The reaction temperature and reaction time of the proteolytic enzyme vary depending on the type of proteolytic enzyme to be added and the plant used as a raw material. However, as an example, the reaction temperature is preferably 30 ° C. to 65 ° C., more preferably 40 ° C. to 65 ° C., still more preferably 45 ° C. to 60 ° C., and preferably 0.05 hours to The reaction time is 24 hours, more preferably 0.2 to 12 hours, still more preferably 0.3 to 6 hours, and particularly preferably 0.5 to 2 hours. .

2. Liquefaction process A liquefaction enzyme is added to the solution obtained through the preparation process, the solution is heated, raised to the reaction temperature of the added liquefaction enzyme, and liquefied for a predetermined reaction time. For this liquefaction reaction, a boiling water bath or a continuous liquefaction apparatus such as a jet cooker can be used.

  The liquefying enzyme to be added is not particularly limited as long as it randomly cleaves the α-1,4 glycosidic bond of starch, and α-amylase (EC 3.2.1.1) can be preferably used. Such a liquefying enzyme may be a commercially available product, and particularly preferably, trade name “Chrytase T10S” (manufactured by Amano Enzyme).

  The amount of liquefied enzyme added is not limited to liquefaction, but is preferably 0.001% to 1.0%, more preferably 0.03% to 0.5% on a mass basis (W / W) relative to the raw material. More preferably, it is 0.1% to 0.4%. If it is 0.001% or more, the liquefaction reaction proceeds sufficiently, and if it is 1.0% or less, it is economical.

  The reaction temperature and reaction time of the liquefaction enzyme vary depending on the type of liquefaction enzyme to be added and the plant used as a raw material. However, as an example, the reaction temperature is preferably 65 ° C. to 120 ° C., more preferably 80 ° C. to 110 ° C., preferably 0.01 to 24 hours, more preferably 0.1 The reaction time is from 12 hours to 12 hours, more preferably from 0.1 to 2 hours.

3. Saccharification Step After the liquefying enzyme solution is lowered to the saccharification enzyme reaction temperature, saccharification enzyme and β-glucan degrading enzyme are added to the solution, and the saccharification reaction is carried out at the reaction temperature for a predetermined reaction time. The solution after the saccharification reaction is treated at a high temperature of 90 ° C. or higher for a predetermined time in order to deactivate the saccharifying enzyme and β-glucan degrading enzyme.

  The saccharifying enzyme to be added may be any saccharifying enzyme as long as dextrins generated by liquefaction are further decomposed into oligosaccharides, and commercially available saccharifying enzymes can be used as appropriate.

  When β-amylase (EC 3.2.1.2) is used as a saccharification enzyme, a solution containing a large amount of maltose which is a disaccharide can be obtained. As such β-amylase, the trade name “β-amylase # 1500S” (manufactured by Nagase ChemteX Corporation) is particularly preferable. And the addition amount of such β-amylase is preferably 0.001% to 2.0%, more preferably 0.1% to 1.5%, still more preferably on a mass basis (W / W) with respect to the raw material. Is 0.2% to 1.0%.

  In addition to β-amylase, it is preferable to further add a so-called debranching enzyme as a saccharifying enzyme. This debranching enzyme is an enzyme for cleaving the α-1,6 bond of dextrins generated by liquefaction, and preferably pullulanase (EC 3.2.1.41) or isoamylase (EC 3.2.1.68). In particular, the trade name “Pulllanase Amano 3” (manufactured by Amano Enzyme) can be used. And the addition amount of such a debranching enzyme is preferably 0.001% to 2.0%, more preferably 0.01% to 1.5%, and still more preferably on a mass basis (W / W) relative to the raw material. Is 0.01% to 1.0%.

  In the present invention, it is desirable that a solution having a low viscosity is finally obtained in consideration of use in foods and beverages. Therefore, it is desirable that the sugar composition contained in the finally obtained solution contains more monosaccharides, disaccharides and the like. Therefore, in addition to β-amylase and debranching enzyme, glucoamylase (EC 3.2.1.3), particularly preferably “Daizyme GPS” (manufactured by Amano Enzyme) is added as a saccharifying enzyme. Can do. And the addition amount of this glucoamylase is preferably 0.001% to 5.0%, more preferably 0.1% to 3.0%, still more preferably 0.00% by mass basis (W / W) relative to the raw material. 2% to 2.5%.

  Furthermore, in the present invention, in consideration of the purpose of obtaining a β-glucan-containing solution having a lower molecular weight, in the saccharification step, in addition to the saccharifying enzyme, a β-glucan degrading enzyme (EC 3.2. It is desirable to add 1.73). As this β-glucan-degrading enzyme, a commercially available one can be used as appropriate, but the product name “Finizym (registered trademark) 250L” (manufactured by Novozymes) can be particularly preferably used.

  The barley pulverized product used in this production method generally contains several millions of β-glucan in terms of weight average molecular weight. Therefore, from the viewpoint of obtaining the final solution viscosity and the desired ability to inhibit micelle formation, β-glucan having a desired weight average molecular weight is obtained by decomposition with β-glucan degrading enzyme. By adjusting the addition amount of the β-glucan degrading enzyme and the pH during the reaction, it is possible to control the weight average molecular weight of β-glucan within a predetermined range.

  In the present invention, β-glucan preferably has a weight average molecular weight of preferably 2,500 to 40,000, more preferably 6,000 to 30,000, still more preferably 7,000 to 25,000, particularly preferably 8. , 20,000 to 20,000. And the addition amount of (beta) -glucan degrading enzyme used for obtaining this weight average molecular weight is 0.0001%-1.0%, More preferably, it is 0.0001%-1.0% by the mass reference | standard (W / W) with respect to a raw material. 001% to 0.1%, more preferably 0.002% to 0.01%. Further, the pH of the solution at the time of the degradation reaction with β-glucan degrading enzyme is not particularly limited as long as the enzyme is not inactivated in a short time, but preferably pH 4.0 to pH 8.0, more preferably pH 4.0. 5 to pH 7.0, more preferably pH 5.0 to pH 6.0.

  The reaction temperature and reaction time in this step vary depending on the type of saccharifying enzyme and β-glucan degrading enzyme to be added and the plant used as a raw material. However, as an example, the reaction temperature is preferably 40 ° C. to 80 ° C., more preferably 50 ° C. to 70 ° C., still more preferably 55 ° C. to 65 ° C., and preferably 0.1 hours to The reaction time is 24 hours, more preferably 0.5 to 12 hours, and still more preferably 1 to 2 hours.

4). Filtration / filter treatment / concentration step The solution obtained through the saccharification step is filtered using diatomaceous earth or activated carbon as an auxiliary agent, and further passed through the filter to obtain a liquid portion from which insoluble portions have been removed. And it concentrates using an evaporator etc. so that it may become Brix according to the use.

  A plant-derived β-glucan-containing syrup can be obtained by the above charging step, liquefaction step, saccharification step, and filtration / filter treatment / concentration step.

<Brix (soluble solid content concentration)>
In the present invention, Brix refers to a soluble solid content concentration (%). The refractive index of an aqueous solution in which the soluble solid content is dissolved is measured at 20 ° C., and provided by ICUMSA (International Commission for Uniform of Sugar Analysis). It is the value converted into the mass / mass percent of the pure sucrose solution based on the conversion table. In this invention, it means the density | concentration of the soluble solid content contained in the aqueous solution, ie, the plant-derived (beta) -glucan containing syrup manufactured at the said manufacturing process. This Brix can be measured by appropriately using a known measurement method that is already known. Generally, a commercially available saccharimeter (for example, a digital refractometer, trade name “RX-5000α”: manufactured by Atago Co., Ltd.) is used. Can be measured.

  The optimal Brix may be appropriately adjusted from the viewpoint of reducing the risk of microbial growth of the syrup while realizing a low viscosity that is easy to handle as a manufactured syrup.

<The ability of β-glucan to inhibit micellization>
Bile acids involved in cholesterol absorption form bile acid micelles when the concentration exceeds a predetermined level. Cholesterol is not absorbed by the small intestine alone, but can be absorbed from the small intestine by being taken into bile acid micelles. That is, cholesterol is absorbed into the small intestine as micelles incorporated into bile acid micelles.

  At this time, β-glucan absorbs cholesterol in the small intestine by inhibiting the association of cholesterol with bile acid micelles, or destroying the micelles once incorporated with cholesterol and releasing cholesterol again. Is inhibited.

  That is, the ability of cholesterol to inhibit micelle formation broadly means that cholesterol is taken into bile acid micelles to form new micelles, thereby inhibiting absorption into the small intestine.

  And it can be confirmed by the following tests whether the plant-derived β-glucan-containing syrup obtained by the above production method has the ability to inhibit cholesterol micellization.

  First, a micelle solution to which other lipids (for example, lecithin, oleic acid, etc.) that can be absorbed in the living body are added in addition to bile acid salt and cholesterol is prepared. Next, β-glucan is extracted from the plant-derived β-glucan-containing syrup obtained by the above production method by a known method. Then, a predetermined amount of the extracted β-glucan solution is added to the prepared micelle solution and incubated at room temperature to 40 ° C. for 12 hours to 48 hours. Then, the incubated solution is divided into an upper layer and a lower layer by centrifugation or the like, and the concentration of free cholesterol contained in the upper layer portion is measured.

  In addition, extraction of β-glucan in plant-derived β-glucan-containing syrup can be appropriately performed using a known method as described above, but as an example, it is an official method of AOAC (Association of Official Analytical Chemistry) International. It is preferable to use the method defined in AOAC 985.12. In addition, a commercially available kit using this method can be used, and examples thereof include a commercially available kit under the trade name “Total Dietary Fiber Assay” (manufactured by Sigma Aldrich).

  In addition, the determination of whether the β-glucan solution extracted by the above method has the ability to inhibit micellization can be qualitatively performed using a sample. For example, when a barley-derived β-glucan-containing syrup is used for the test, a standard-derived β-glucan solution obtained by decomposing the obtained barley β-glucan standard with a β-glucan degrading enzyme is prepared. Then, after adding a predetermined amount of standard-derived β-glucan solution to the prepared micelle solution, the concentration of free cholesterol is measured through incubation, centrifugation, and the like in the same manner as in the above test. The above test is performed using the barley β-glucan sample thus obtained and the β-glucan derived from the above production method, and the micelle formation inhibitory ability is evaluated by comparing the free cholesterol concentration.

<Ability to suppress blood sugar level rise>
The ability of the plant-derived β-glucan-containing syrup according to the present invention to suppress an increase in blood glucose level in animals (preferably humans) can be confirmed by various methods. As an example, first, a healthy person who is sufficiently hungry ingests a predetermined amount of the plant-derived β-glucan-containing syrup as the first meal (first meal) according to the present invention, and then measures the blood glucose level at regular intervals. Can be confirmed.

  Furthermore, after a predetermined time, the next meal (second meal) was ingested, and the effect of the plant-derived β-glucan-containing syrup ingested as the first meal on the blood glucose level increase by the next meal (second meal) was determined at regular intervals. To evaluate. Thereby, it is possible to confirm the second meal effect which means that the meal taken as the first meal also affects the blood glucose level after the next meal (second meal).

<Plant-derived β-glucan-containing syrup>
The plant-derived β-glucan-containing syrup according to the present invention desirably has the following characteristics. That is, the weight average molecular weight is preferably 2,500 to 40,000, more preferably 6,000 to 30,000, still more preferably 7,000 to 25,000, and particularly preferably 8,000 to 20,000. It is preferable to contain β-glucan. Thereby, for example, not only the handling as a raw material for foods and beverages is facilitated, but also the physiological function of β-glucan can be sufficiently exhibited.

  For example, from the viewpoint of handling as a raw material for foods and beverages, the viscosity of the plant-derived β-glucan-containing syrup is preferably 20,000 cP or less, more preferably 1,000 cP to 15,000 cP, and still more preferably 2,000 cP to It is desirable to be 10,000 cP. The Brix of the plant-derived β-glucan-containing syrup is preferably 30% to 80%, more preferably 50% to 80%, and still more preferably 60% to 75%. The pH of the plant-derived β-glucan-containing syrup is preferably pH 4.0 to pH 8.0, more preferably pH 4.5 to pH 7.0, and still more preferably pH 5.0 to pH 6.0. .

<Food or beverage containing plant-derived β-glucan-containing syrup>
The plant-derived β-glucan-containing syrup having the above characteristics can be used as a material for the food or beverage by adding it to various foods or beverages. Examples of this food include fruits (cut fruits such as apples, bananas and melons, pickled syrup, etc.), processed cereals (bread, strawberries, etc.), processed meats (ham, sausages, etc.), dairy products (butter, Cheese, yogurt, etc.), processed fruits (jam, marmalade, etc.), confectionery (chocolate, cookies, cakes, jelly, etc.), seasonings (soy sauce, sauce, mirin, etc.), supplements, etc. . Examples of beverages include various beverages such as beer, non-alcoholic beverages, juice, coffee, tea, green tea, and carbonated beverages.

  In addition to the plant-derived β-glucan-containing syrup, various components can be added to these foods or beverages depending on the type of the food. For example, sugars such as glucose, fructose, sucrose, maltose, starch syrup, and lactose, sugar alcohols such as sorbitol, erythritol, maltitol, and xylitol, high-intensity sweeteners such as aspartame, stevioside, sucralose, and acesulfame K Organic acids such as citric acid, tartaric acid, malic acid, succinic acid, lactic acid, L-ascorbic acid, dl-α-tocopherol, vitamin B, vitamins such as nicotinic acid amide, calcium pantothenate, glycerin fatty acid ester, polyglycerin Surfactants such as fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, thickeners such as gum arabic, carrageenan, pectin, agar, stabilizers such as casein, gelatin, amino acids, calcium salts Minine Le acids, sodium erythorbate, glycerol, propylene glycol and the like additives, dyes, perfumes, include food or beverage material, such as preservatives.

Moreover, the plant-derived β-glucan-containing syrup added to food or beverage may be used as a product as it is as a syrup, but may be contained in food or beverage,
The amount added is preferably 0.01% to 100%, more preferably 0.1% to 50%, and still more preferably 1% to 20% based on the mass of the food or beverage.

  EXAMPLES Hereinafter, although an Example demonstrates this invention in more detail, this invention is not limited to them. In the following examples, “%” means mass / mass% unless otherwise specified.

[Example 1]
In Example 1, the following materials and methods were used.
<Preparation of plant-derived β-glucan-containing syrup>
After being dispersed with pure water so that the pulverized barley was 30%, it was stirred until it was completely dispersed in the pure water while the barley pulverized mass disappeared while heating to 50 ° C. in a thermostatic bath. Thereafter, 10% calcium hydroxide solution was added to adjust the pH of the barley grind solution to 5.6. Subsequently, 0.05% of the barley pulverized product solution with a trade name “Sumiteam P” (manufactured by Shin Nippon Chemical Industry Co., Ltd.) as a proteolytic enzyme (EC 3.4.22.2) based on the mass of the raw material (W / W) , As the liquefying enzyme α-amylase (EC 3.2.1.1), the trade name “Chrytase T10S” (manufactured by Amano Enzyme) was added at a concentration of 0.4% on a mass basis (W / W) relative to the raw material And reacted at 50 ° C. for 1 hour.

  Next, the reaction solution was placed in a boiling water bath and heated to 90 ° C. or higher to cause a liquefaction reaction for 1 hour. After the liquefaction reaction, the added liquefied enzyme was inactivated by autoclaving at 120 ° C. for 15 minutes.

  Next, the reaction solution is cooled to 60 ° C. in a thermostatic bath, and the product name “β-amylase # 1500S” (manufactured by Nagase ChemteX Corporation), which is β-amylase (EC 3.2.1.2) as a saccharifying enzyme. At a concentration of 0.52% based on the mass of the raw material (W / W), and the product name “Pulllanase Amano 3” (manufactured by Amano Enzyme), which is pullulanase (EC 3.2.1.41), based on the mass based on the raw material ( The product name “Dyzyme GPS” (manufactured by Amano Enzyme), which is glucoamylase (EC 3.2.1.3), at a concentration of 0.26% in terms of W / W) is 0 on a mass basis (W / W) relative to the raw material. At a concentration of 26%, the trade name “Finzym (registered trademark) 250L” (manufactured by Novozymes), which is β-glucan-degrading enzyme (EC 3.2. In 0052% of concentration was added into the reaction solution. Then, after saccharification reaction was performed at 60 ° C. for 2 hours, the enzyme was inactivated by heating to 90 ° C.

  Next, 80 ° C. to 85 ° C. is applied to Nutsche covered with filter paper (trade name “Qualitative filter paper No. 2”: ADVANTEC) covered with diatomaceous earth (trade name “Radiolite # 500S”: manufactured by Showa Chemical Industry Co., Ltd.). The reaction solution held in was passed through. The filtrate was held at 70 ° C. to 75 ° C. through a Nutsche with a membrane filter (trade name “Mixed Cellulose Esters (MCE) Membrane Filters” manufactured by ADVANTEC) having a pore size of 0.8 μm. Liquid. The obtained filtrate was concentrated to such an extent that it was not fixed by an evaporator. Here, the term “fixed” refers to a state in which part of the syrup on the bottom of the flask loses fluidity and adheres to the flask during concentration.

<Method for quantifying β-glucan>
Β-glucan in the plant-derived β-glucan-containing syrup prepared by the above method was determined using the McCleary method (trade name “MIXED LINKAGE BETA-GLUCAN ASSAY KIT: manufactured by Megazyme) using the McCleary method ( That is, the plant-derived β-glucan-containing syrup adjusted to 10% Brix was added to 5 ml in a 15 ml tube, and 2.5 g of finely pulverized ammonium sulfate was added and dissolved. The supernatant was removed by centrifugation at 3,000 rpm for 10 minutes at 4 ° C. 1 mL of 50% (V / V) aqueous ethanol solution was added to the remaining pellet, and the pellet was suspended by vigorous stirring. (V / V) 10 mL of ethanol aqueous solution was added and mixed. The supernatant was removed by centrifuging at rpm for 5 minutes, and the operations of pellet suspension, ethanol addition, centrifugation and supernatant removal were repeated again, and the pellet was washed with 20 mM sodium phosphate buffer (pH 6.5). After redissolving in 8 mL, 200 μL of the lichenase solution was added to 4.8 mL of the solution diluted 10-fold with the above buffer, and incubated at 40 ° C. for 5 minutes. 100 μL of β-glucosidase solution was added to the tube and reacted at 40 ° C. for 15 minutes, after which 3 mL of glucose oxidase / peroxidase (GOPOD) was added to the tube, reacted at 40 ° C. for 20 minutes, and absorbance at 510 nm. According to the method described in the kit, the absorbance of 100 μg of glucose was separately obtained. Measured, as a reference. In addition, .Beta- glucan concentrations as blank plus 50mM acetate buffer (pH 4.0) 100 [mu] L in place of β- glucosidase solution was determined by the following equation.

β-glucan concentration (% with respect to soluble solid content) = ΔA × F × 9 × D × (100 / Brix)
here,
ΔA = absorbance of the sample−absorbance of the blank F = 100 / absorbance of 100 μg of glucose D = dilution ratio when the syrup was diluted

<Method for Extracting β-Glucan and Method for Measuring Its Molecular Weight>
The weight average molecular weight of β-glucan in the plant-derived β-glucan-containing syrup prepared by the above method was measured by the following method. First, based on the official method of AOAC985.29, it refine | purified using the measurement kit (Brand name "Total Dietary Fiber Assay": Sigma Aldrich company make). That is, 1 g of plant-derived β-glucan-containing syrup and 50 mL of 0.08M phosphate buffer (pH 6.0) in an Erlenmeyer flask, and 100 μL of α-amylase (EC 3.2.1.1) solution attached to the above kit After adding and capping with aluminum foil, it was incubated in boiling water for 30 minutes. Thereafter, after cooling at room temperature for 30 minutes, about 10 mL of 0.275N sodium hydroxide aqueous solution was added to the reaction solution to adjust the pH to 7.5 ± 0.2. Thereafter, 100 μL of a solution obtained by diluting the protease (EC 3.4.21.62) solution attached to the kit to 50 mg / mL is added to the reaction solution, and incubated at 60 ° C. for 30 minutes with shaking. Cooling for a minute was performed. Next, 10 mL of a 0.325N hydrochloric acid aqueous solution was added to the reaction solution to adjust the pH to 4.3 ± 0.3. Thereafter, 100 μL of the amyloglucosidase (EC 3.2.1.3) solution attached to the kit was added to the reaction solution, and incubated at 60 ° C. for 30 minutes with shaking. Thereafter, the reaction solution was boiled for 3 minutes to deactivate the enzyme. Thereafter, the reaction solution was passed through filter paper (trade name “Qualitative Filter Paper No. 2” manufactured by ADVANTEC).

  Next, a cation exchange resin (trade name “Diaion PA218”: manufactured by Mitsubishi Chemical Corporation) and an anion exchange resin (trade name “Diaion PA408”: manufactured by Mitsubishi Chemical Corporation) are mixed and stirred in the reaction solution. The solution was passed through a solid layer extraction column (trade name “Sep-Pak C18 Cartridge” manufactured by Waters). 5 mL of the reaction solution allowed to flow was taken, 2.5 g of ammonium sulfate was added thereto, and the mixture was allowed to stand at 4 ° C. for 20 hours. Then, after centrifuging at 3,000 rpm for 10 minutes at 4 ° C., the supernatant was discarded, and 5 mL of 75% ethanol was added to the pellet and washed using a vortex mixer. Then, after centrifuging again at 3,000 rpm for 10 minutes at 4 ° C., the supernatant was discarded, and 5 mL of 75% ethanol was added to the pellet and washed using a vortex mixer. After repeating the centrifugation and washing steps again, pure water was added to the obtained pellets and dissolved by heating at 80 ° C. or higher. The solution was passed through a membrane filter having a pore size of 0.45 μm (trade name “MILLEX (registered trademark) -HP 0.45 μm”: manufactured by MILLIPORE). The molecular weight distribution and weight average molecular weight of the β-glucan solution were measured by gel filtration chromatography.

The molecular weight distribution and the weight average molecular weight were measured by gel filtration chromatography using HLC-8220GPC (manufactured by Tosoh Corporation) under the following conditions.
Column: Three connected columns of G2500PW-G3000PW-G6000PW (manufactured by Tosoh Corporation) Solvent: Pure water Temperature: 80 ° C
Flow rate: 1.0 ml / min
Detection: RI (differential refractive index)
The obtained molecular weight distribution and weight average molecular weight were calculated based on a calibration curve prepared using pullulan having a known molecular weight (trade name “STANDARD P-82” manufactured by Shodex).

<Viscosity measurement method>
1.2 mL each of plant-derived β-glucan-containing syrup obtained by the above production method is put on a plate of an E-type viscometer (trade name “VISCONIC ED”: manufactured by Tokyo Keiki Co., Ltd.), and the viscosity of the syrup in each Brix is measured. It was measured. The temperature during measurement was 25 ° C.

<Characteristic evaluation results of β-glucan in plant-derived β-glucan-containing syrup>
As Example 1, a plant-derived β-glucan-containing syrup obtained by the above production method was prepared. And each characteristic of the density | concentration, molecular weight distribution, and weight average molecular weight of Brix of the syrup which concerns on this Example 1, and the extracted (beta) -glucan was evaluated.

[Comparative Example 1]
As Comparative Example 1, special barley flour (β-glucan content is 7.5% by mass) is used as a raw material, and the trade name “Dyzyme” which is glucoamylase (EC 3.2.1.3) described in the above production method. Instead of “GPS” (manufactured by Amano Enzyme), the trade name “transglucosidase L Amano” (manufactured by Amano Enzyme), which is transglucosidase (EC 3.2.1.20) as a transferase, is used as a mass standard (W / A plant-derived β-glucan-containing syrup obtained by the same method as in the above production method except that 0.1% was added in (W). Here, the β-glucan-containing syrup produced in Comparative Example 1 was fixed at a Brix of 30% or less in the concentration step. And each characteristic of Brix of the syrup which concerns on the comparative example 1, and the density | concentration of the extracted (beta) -glucan, molecular weight distribution, and a weight average molecular weight was evaluated. The molecular weight distribution and the weight average molecular weight were measured using a high performance liquid chromatograph Prominence (manufactured by Shimadzu Corporation) under the following gel filtration chromatography conditions.
<Measurement conditions>
Column: GS520-GS320 (manufactured by Shodex)
Solvent: Pure water Temperature: 50 ° C
Flow rate: 1.0 ml / min
Detection: CAD (corona charged particle detector: manufactured by Nippon Dionex)

  First, FIG. 1 is a diagram showing the results of extracting β-glucan from the plant-derived β-glucan-containing syrup obtained in Example 1 and measuring the molecular weight distribution by gel filtration chromatography. FIG. 2 is a diagram showing the results of extracting β-glucan from the plant-derived β-glucan-containing syrup obtained in Comparative Example 1 and measuring the molecular weight distribution by gel filtration chromatography. Table 1 shows how much β-glucan of which molecular weight is present based on the molecular weight distribution of FIG.

  According to this, the plant-derived β-glucan-containing syrup obtained by the production method according to Example 1 contains a large amount of β-glucan having a molecular weight of 2,000 to 10,000 and β-glucan having a molecular weight of less than 2,000. It was possible to obtain a large amount of low-molecular β-glucan by the above production method.

  Table 2 shows the results of comparing the characteristics of Example 1 and Comparative Example 1 of the weight average molecular weight, the β-glucan concentration, and the maximum Brix that can exist as a syrup. In general, when Brix is increased, the syrup becomes less fluid and the syrup cannot be handled substantially. The Brix indicates the maximum value of Brix having fluidity that can be handled as a syrup. Specifically, the presence or absence of fixation during concentration was used as an index.

  According to Table 2, the plant-derived β-glucan-containing syrup obtained by the production method according to Example 1 contains β-glucan at a concentration of 4.1% with respect to the soluble solid content. I understood. Moreover, it turned out that the weight average molecular weight is also 13,000, and is a sufficiently low value compared with the weight average molecular weight of the comparative example 1. In general, if Brix is low, the risk of microbial growth increases and is not suitable for use as a food or beverage. On the other hand, however, increasing Brix increases the viscosity of the syrup, making it difficult to handle. From this point of view, the plant-derived β-glucan-containing syrup of Example 1 has a sufficiently low weight average molecular weight, and is a syrup having an appropriate viscosity even with a high Brix of Brix of 70% or more. Use as a beverage is expected.

<Results of characterization of plant-derived β-glucan-containing syrup>
For the syrup according to Example 1, in addition to the above characteristics, characteristics such as the composition ratio of monosaccharides, disaccharides, trisaccharides, saccharides higher than tetrasaccharides, and viscosity of the syrup were also evaluated. The method for evaluating the sugar composition is as follows. The syrup was diluted to about Brix 10%, and an appropriate amount of activated carbon (trade name “Shirakaba A” manufactured by Nippon Enviro Chemicals) was added and mixed. Next, the sample solution was heated until boiling, and passed through a filter paper (trade name “qualitative filter paper No. 2” manufactured by ADVANTEC). Thereafter, a cation exchange resin (trade name “Diaion PA218”: manufactured by Mitsubishi Chemical Corporation) and an anion exchange resin (trade name “Diaion PA408”: manufactured by Mitsubishi Chemical Corporation) are mixed and stirred in the sample solution. The solution was passed through a layer extraction column (trade name “Sep-Pak C18 Cartridge” manufactured by Waters). The Brix of the sample is adjusted to about 1% to 5%, and is passed through a membrane filter (trade name “MILLEX (registered trademark) -HP 0.45 μm”: Merck) having a pore size of 0.45 μm, and sugar A sample for composition analysis was obtained. The sugar composition was analyzed using a trade name “Alliance (registered trademark) HPLC system” (manufactured by Nihon Waters).
<Measurement conditions>
Column: ULTRON PS80-N (manufactured by Shimadzu LLC)
Solvent: Pure water Temperature: 60 ° C
Flow rate: 0.6 ml / min
Detection: RI (differential refractive index)

[Example 2]
Moreover, as Example 2, the plant-derived β produced by the same method except that the glucoamylase to be added is 0.052% on a mass basis (W / W) with respect to the raw material from the production method according to Example 1 above. -A glucan-containing syrup was prepared. And about the syrup which concerns on this Example 2, similarly to Example 1, characteristics, such as a monosaccharide, a disaccharide, a trisaccharide, the composition ratio of saccharides more than a tetrasaccharide, the viscosity of a syrup, were evaluated.

[Example 3]
Moreover, as Example 3, all the plant-derived β produced by the same method except that the glucoamylase to be added was 2.08% by mass standard (W / W) with respect to the raw material from the production method according to Example 1 above. -A glucan-containing syrup was prepared. And about the syrup which concerns on this Example 2, similarly to Example 1, characteristics, such as a monosaccharide, a disaccharide, a trisaccharide, the composition ratio of saccharides more than a tetrasaccharide, the viscosity of a syrup, were evaluated.

[Example 4]
Moreover, as Example 4, the plant-derived β-glucan-containing syrup was prepared by the same method except that glucoamylase was not added from the production method according to Example 1 above. And about the syrup which concerns on this Example 2, similarly to Example 1, characteristics, such as a monosaccharide, a disaccharide, a trisaccharide, the composition ratio of saccharides more than a tetrasaccharide, the viscosity of a syrup, were evaluated.

  Table 3 shows the results of comparing the characteristics such as monosaccharide, disaccharide, trisaccharide, composition ratio of saccharides higher than tetrasaccharide, syrup viscosity, and the like for Examples 1 to 4.

  As apparent from Table 3 above, the plant-derived β-glucan-containing syrups according to Examples 1 to 4 all have β-glucan having a weight average molecular weight in the range of 2,500 to 40,000. Was included. The plant-derived β-glucan-containing syrup of Examples 1 to 4 containing β-glucan having a weight average molecular weight of 2,500 to 40,000 is 3.8% to 4.2% with respect to the soluble solid content. Β-glucan concentration.

  In general, it is desirable that the syrup has a Brix of 70% or more from the viewpoint of reducing the risk of microbial growth during storage, for example. On the other hand, a syrup having such a high Brix has a high viscosity. It is unsuitable for use as a food or beverage.

  However, any of the plant-derived β-glucan-containing syrups of Examples 1 to 4 containing β-glucan having a weight average molecular weight of 2,500 to 40,000 is an appropriate viscosity as the viscosity when Brix is 70% or more. It was found to have In particular, since the plant-derived β-glucan-containing syrup obtained in Examples 1 to 3 was added with glucoamylase in the saccharification step, the plant-derived β obtained in Example 4 in which it was not added. -The proportion of monosaccharides in the sugar composition is large compared to the glucan-containing syrup. Therefore, with respect to the plant-derived β-glucan-containing syrup of Example 4, the plant-derived β-glucan-containing syrup of Examples 1 to 3 has a viscosity as low as 3,100 cP to 9,900 cP. As a result, it has been found that it has excellent properties as a raw material for foods and beverages.

  Moreover, FIG. 3 is a figure which shows the result of having measured the relationship between each Brix and viscosity in the plant-derived β-glucan-containing syrup of Example 1. According to this, even if Brix explained above is 70% or more, it has an appropriate viscosity of 10,000 cP or less.

  Furthermore, for example, when considering application to the use as a raw material for foods and beverages, naturally, the plant-derived β-glucan-containing syrup of Examples 2 to 4 is sufficiently utilized as a raw material for foods and beverages. However, it is particularly preferable to use the plant-derived β-glucan-containing syrup of Example 1.

  Since the plant-derived β-glucan-containing syrup of Example 3 to which a larger amount of glucoamylase was added in the saccharification step contains a large amount of monosaccharides, there is a possibility that saccharides may precipitate when Brix is increased. In addition, the plant-derived β-glucan-containing syrup of Example 2 in which a smaller amount of glucoamylase was added in the saccharification process and the plant-derived β-glucan-containing syrup of Example 4 in which no glucoamylase was added at all was used The viscosity is slightly higher than that of the plant-derived β-glucan-containing syrup of Example 1 because it contains a large amount of disaccharides or more.

  Therefore, the plant-derived β-glucan-containing syrup of Example 1 to which glucoamylase in the range of 0.2% to 2.5% is added in the saccharification step has an appropriate viscosity, and is used as a raw material for foods and beverages. It was found to be particularly excellent as a use for.

<Evaluation method of micellization inhibition ability>
Evaluation of the ability of plant-derived β-glucan-containing syrup obtained by the above production method to inhibit the micellization of cholesterol is described in Nagaoka et al. The Journal of Nutrition (1999), vol. 129, pp. The method described in 1725-1730 was appropriately modified. Specifically, it was performed through the following steps.
1. Preparation of micelle solution The micelle solution used for evaluating the ability of plant-derived β-glucan-containing syrup obtained by the above production method to inhibit the micellization of cholesterol was prepared as follows.

  The micelle solution was prepared in a glass bottle so that 250 mL of 15 mM phosphate buffer (pH 7.4) was added and each reagent described in Table 4 was adjusted to a predetermined final concentration. The glass bottle containing the solution was placed in an ultrasonic cleaner (trade name “UT-604”: manufactured by Sharp Corporation), and the reagent was completely dispersed with ultrasonic waves for 30 minutes.

2. Extraction method of β-glucan from plant-derived β-glucan-containing syrup It was described in the above section “Method for extracting β-glucan and method for measuring its molecular weight” from the plant-derived β-glucan-containing syrup prepared in Example 1. Β-glucan was extracted using the method. The pellet of β-glucan after ammonium sulfate precipitation and ethanol washing was stored at −80 ° C. for about 2 hours and frozen. Thereafter, the pellets were freeze-dried for 20 hours or more at a degree of vacuum of 0.133 mBar or less by a freeze dryer (trade name “FreeZone (registered trademark)” manufactured by Labconco).

3. Preparation of standard-derived β-glucan solution A barley-derived β-glucan standard (manufactured by Sigma Aldrich) was dissolved in pure water to prepare a plurality of 1% solutions. Next, lichenase (EC 3.2.1.73: manufactured by Megazyme) was added to each solution of 0.125 and 1.0 U, and the reaction was performed at 38 ° C. for a time determined for each solution. Immediately after the elapse of time, the enzyme was inactivated by heating at 90 ° C. for 10 minutes. Table 5 shows the amount of enzyme and reaction time used in each example. In addition, each obtained reaction liquid measured the molecular weight distribution by the gel filtration chromatography. The measurement was performed by the same method as that described in the section “Method for extracting β-glucan and method for measuring its molecular weight”.

4). Evaluation test of ability to inhibit micellization Enzymatic degradation of β-glucan lyophilized powder 2.5 mg extracted from plant-derived β-glucan-containing syrup and suspended in 0.25 g of pure water and β-glucan preparation by the above method 0.25 g of the 1% β-glucan solution was put into a 15 mL tube, and 4.75 g of the micelle solution was added to make the final concentration of β-glucan 0.05% (W / W). At this time, as a positive control, a suspension of 2.5 mg of barley-derived β-glucan preparation (manufactured by Sigma Aldrich), which has been confirmed to inhibit cholesterol absorption, was suspended in 0.25 g of pure water as a micelle solution. Mixed. As a negative control, 2.5 mg of glucose suspended in 0.25 g of pure water was mixed with a micelle solution (Comparative Example 2). Moreover, 0.25 g of pure water was mixed with the micelle solution as a blank (Comparative Example 3).

  Thereafter, each solution was completely dispersed by ultrasonic treatment for 30 minutes with an ultrasonic cleaner (trade name “UT-604”: manufactured by Sharp Corporation), and then incubated at 37 ° C. for 24 hours. Thereafter, each reaction solution was 20,000 rpm (100,000 g) using a swing rotor (trade name “SW28”: manufactured by Beckman Coulter) and a centrifuge (trade name “L7-55”: manufactured by Beckman Coulter). For 60 minutes. 200 μL of the upper layer of each reaction solution was collected, and 3 mL of a coloring solution (trade name “Free Cholesterol E-Test Wako” manufactured by Wako Pure Chemical Industries, Ltd.) was added to the collected solution. The solution after addition was incubated at 37 ° C. for 5 minutes, and then the absorbance was measured at 600 nm using a spectrophotometer (trade name “BioSpec-mini” manufactured by Shimadzu Corporation) to calculate the free cholesterol concentration.

[Example 5]
As Example 5, using β-glucan extracted from the plant-derived β-glucan-containing syrup by the method described in the above section “2. Extraction method of β-glucan from plant-derived β-glucan-containing syrup”, The micelle inhibition ability was evaluated by the method described in the section “4. Evaluation test of micelle inhibition ability”.

[Example 6] to [Example 11]
In addition, as Example 6, according to the method described in the above section “3. Preparation of β-glucan solution derived from sample”, the amount of barium-derived β-glucan sample was added to the lichenase addition amount and reaction described in Table 5 By reacting with lichenase over time, β-glucan having the weight average molecular weight shown in Table 5 was also prepared. Then, using this β-glucan, the micelle inhibition ability was evaluated by the method described in the section “4. Evaluation test of micelle inhibition ability”. Similarly, as Examples 7 to 11, each β-glucan having the weight average molecular weight described in Table 5 was prepared with the addition amount and reaction time described in Table 5, and micelle formation in each β-glucan was performed. Inhibitory ability was evaluated.

[Reference Example 1] to [Reference Example 3]
As Reference Example 1, according to the method described in the above section “3. Preparation of β-glucan solution derived from sample”, β-glucan sample derived from barley was added with the amount of lichenase and reaction time shown in Table 5. By reacting with lichenase, β-glucan having a larger weight average molecular weight than that of β-glucan used in Examples 5 to 11 was prepared. The specific weight average molecular weight is as shown in Table 5. Then, using this β-glucan, the ability of β-glucan to inhibit micellization was evaluated. Similarly, as Reference Example 2 and Reference Example 3, each β-glucan having the weight average molecular weight described in Table 5 was also prepared with the addition amount and reaction time described in Table 5, and each β-glucan The ability to inhibit micellization was evaluated.

[Comparative Example 2] and [Comparative Example 3]
Furthermore, as described in Comparative Example 2, as described in “4. Evaluation test of micelle inhibition ability”, instead of β-glucan, 2.5 mg of glucose suspended in 0.25 g of pure water was micellized. It used for the evaluation test of inhibition ability. Further, as Comparative Example 3, 0.25 g of pure water was used in the evaluation test of micelle inhibition ability in place of β-glucan.

  Table 5 evaluated each micellization inhibitory ability in Examples 5 to 11 and Reference Examples 1 to 3, Comparative Example 2 and Comparative Example 3 based on the free cholesterol concentration (mg / dL). Results are shown.

  According to Table 5, in the β-glucan extracted from the plant-derived β-glucan-containing syrup of Example 5 having a smaller weight average molecular weight (13,000), Comparative Example 2 and Comparative Example 3 used as negative controls In contrast, a higher free cholesterol concentration (59.4 mg / dL) was exhibited. Similarly, the β-glucan according to the present invention preferably has a weight average molecular weight of 2,500 to 40,000, and the β-glucan having the weight average molecular weights of Examples 6 to 11 is also a negative control. As compared with Comparative Example 2 and Comparative Example 3 used as above, a higher free cholesterol concentration was shown.

  In addition, it has been reported that β-glucan having a very large weight average molecular weight (for example, Reference Example 3) has the ability to inhibit micellization. This is consistent with the high free cholesterol concentration measured in Reference Example 3. On the other hand, in Example 5 to Example 11, compared with Reference Example 1 and Reference Example 2 whose weight average molecular weight is larger than that of β-glucan of Example 5 to Example 11, equivalent or more free cholesterol. Concentration was indicated.

  That is, as in the present invention, even β-glucan having a weight average molecular weight of 2,500 to 40,000 has a very high weight average molecular weight (300,000 or more) as conventionally reported. Similar to β-glucan, it was suggested that cholesterol could be taken up by bile acid micelles to form new micelles and be absorbed by the small intestine.

<Evaluation test of blood glucose level rise suppression ability>
1. Preparation of plant-derived β-glucan (weight average molecular weight: 12,000) -containing syrup In preparation of plant-derived β-glucan (weight-average molecular weight: 12,000) -containing syrup, a barley pulverized product solution as a raw material is used as a barley pulverized product. Was prepared according to the method for preparing a plant-derived β-glucan-containing syrup of Example 1 except that it was prepared by dispersing in pure water so as to be 20% (in Example 1, 30% of barley grind was 30%) To be mixed with pure water). Finally, a plant-derived β-glucan-containing syrup (weight average molecular weight: 12,000, 4.0% by mass based on the entire soluble solid content) was obtained.

2. Preparation of plant-derived β-glucan-free syrup In preparation of plant-derived β-glucan-free syrup, the barley pulverized product solution as a raw material was dispersed in pure water so that the barley pulverized product was 20%. The addition amount of the prepared point and the trade name “Finzym (registered trademark) 250L” (manufactured by Novozymes), which is β-glucan degrading enzyme (EC 3.2.1.73), is 10 times that of the preparation method of Example 1 above. It was prepared according to the preparation method of Example 1 above, except that it was added to the reaction solution in an amount (concentration of 0.052% by mass basis (W / W) relative to the raw material). And finally, the syrup which does not contain plant origin beta-glucan (0.2 mass% with respect to the whole soluble solid content) was obtained.

3. Preparation of plant-derived β-glucan (weight average molecular weight: 81,000) -containing syrup In the preparation of plant-derived β-glucan (weight average molecular weight: 81,000) -containing syrup, the barley pulverized product solution used as a raw material was crushed with barley. The product name is “Finzym (registered trademark) 250L” (manufactured by Novozymes), which is a β-glucan-degrading enzyme (EC 3.2.1.73). All of the above examples except that it was added to the reaction solution in an amount 1/2 times that of the preparation method of Example 1 above (concentration of 0.0026% on a mass basis (W / W) relative to the raw material). Prepared according to one preparation method. Finally, a plant-derived β-glucan-containing syrup (weight average molecular weight: 81,000, 3.9% by mass based on the entire soluble solid content) was obtained.

[Example 12] and [Example 13]
Two healthy subjects (both human) were prepared as 1 above according to the following test method as subjects (subject A: NGSP value of HbA1c 5.1%, subject B: NGSP value of HbA1c 5.1%). An evaluation test was conducted on the ability to suppress increase in blood glucose level of plant-derived β-glucan (weight average molecular weight: 12,000) -containing syrup. In addition, the result of the evaluation test by the subject A was set as Example 12, and the result of the evaluation test by the test subject B was set as Example 13.

  First, each subject was refrained from eating and drinking other than water after 9:00 pm on the day before the test. Next, after measuring the fasting blood glucose level in the early morning, the plant-derived β-glucan (weight average molecular weight: 12,000) -containing syrup prepared in 1 above was used as the first meal (first meal), and carbohydrates 50 g The amount of saccharide contained 2.1 g of β-glucan). And after the said intake, the blood glucose level in 30 minutes, 60 minutes, 90 minutes, 120 minutes, and 150 minutes was measured, respectively. Furthermore, after measuring the blood glucose level on an empty stomach before lunch (240 minutes after ingesting syrup), 150 g of rice (52 g of carbohydrates) was ingested as a second meal, and after syrup ingestion at 270 minutes, 300 minutes, 330 minutes, and 360 minutes Each blood glucose level was measured. In the above evaluation test, the blood glucose level was measured using an enzymatic method (hexokinase method) and an automatic analyzer (manufactured by Beckman Coulter). During the evaluation test, the subject was allowed to remain seated. The measurement results of blood glucose levels obtained from two healthy subjects by the above evaluation test are shown in Table 6 and FIGS. 4 and 5, respectively.

[Comparative Example 4] and [Comparative Example 5]
Assuming that the same two healthy subjects as in Examples 12 and 13 were subjects (Subject A and Subject B), 50 g of syrup prepared in 2 above for each subject (0.1 g of β-glucan in the carbohydrate) And an evaluation test of the ability to suppress an increase in blood glucose level. And the result of the evaluation test by the subject A is shown in Comparative Example 4 and the result of the evaluation test by the subject B is shown as Comparative Example 5 in Table 6 and FIGS. 4 and 5 respectively.

  The test methods in Comparative Examples 4 and 5 were the same as those in Examples 12 and 13 except that the syrup containing no plant-derived β-glucan prepared in 2 was used as the syrup to be ingested by each subject. The test was performed in the same manner as in the above. Moreover, the said evaluation test was done for a sufficient period (5 days or more) from the test implementation date of the said Example 12 and Example 13.

[Comparative Example 6] and [Comparative Example 7]
Assuming that the same two healthy subjects as in Examples 12 and 13 were subjects (Subject A and Subject B), 50 g of syrup prepared in 3 above for each subject (2.0 g of β-glucan in the carbohydrate) And an evaluation test of the ability to suppress an increase in blood glucose level. And the result of the evaluation test by the subject A is shown in Comparative Example 6 and the result of the evaluation test by the subject B is shown as Comparative Example 7 in Table 6 and FIGS. 4 and 5 respectively.

  The test methods in Comparative Example 6 and Comparative Example 7 were the same as those described above except that the plant-derived β-glucan (weight average molecular weight: 81,000) -containing syrup prepared in 3 was used as the syrup to be ingested by each subject. The test was performed in the same manner as the test methods in Example 12 and Example 13. Moreover, the said evaluation test was done for a sufficient period (5 days or more) from the test implementation day of the said Example and comparative example.

  4 and 5 are diagrams showing the evaluation test results of the ability to suppress the increase in blood glucose level of each syrup taken by subject A or subject B, respectively. Table 6 shows specific blood glucose levels obtained in an evaluation test of the ability to suppress the increase in blood glucose level in each syrup ingested by subjects A and B. According to this, in both subject A and subject B, when a plant-derived β-glucan (weight average molecular weight 12,000) -containing syrup is ingested as a first meal on an early morning fast (Example 12 and Example 13) As compared with the case where syrup not containing plant-derived β-glucan was ingested as a first meal (Comparative Example 4 and Comparative Example 5), the increase in blood glucose level was significantly suppressed. Furthermore, even after ingesting cooked rice as the second meal, the plant-derived β-glucan (weight average molecular weight 12,000) -containing syrup is ingested as the first meal (Example 12 and Example 13). Compared with the case where syrup containing no derived β-glucan was ingested (Comparative Example 4 and Comparative Example 5), the suppression of the increase in blood glucose level was sustained, and the so-called second-meal effect was confirmed.

  In addition, in Comparative Example 6 and Comparative Example 7 in which plant-derived β-glucan (weight average molecular weight 81,000) -containing syrup was ingested, as in Example 12 and Example 13, no plant-derived β-glucan was contained. As compared with Comparative Example 4 and Comparative Example 6 in which syrup was ingested, it was confirmed that blood glucose level was inhibited from rising. However, plant-derived β-glucan (weight average molecular weight 12,000) -containing syrup having a smaller weight average molecular weight than that used in Example 12 and Example 13 than plant-derived β-glucan (weight average molecular weight 81,000) -containing syrup. As is confirmed in Examples 1 to 4 and Comparative Example 1, this has a high Brix, so the risk of microbial growth is low and has an appropriate viscosity, which is more desirable in food and beverage applications.

  According to the present invention, it is possible to provide a plant-derived β-glucan-containing syrup having a sufficiently low viscosity and capable of inhibiting the micellization of cholesterol and suppressing the increase in blood glucose level. Therefore, the present invention can be used in the food and beverage industries. It is.

Claims (8)

  A plant-derived β-glucan-containing syrup comprising a plant-derived β-glucan having a weight average molecular weight of 2,500 to 40,000 in a content of 2% by mass to 8% by mass with respect to the entire soluble solid content.   The plant-derived β-glucan-containing syrup according to claim 1, wherein the plant-derived β-glucan contained has an ability to inhibit cholesterol micellization.   The plant-derived β-glucan-containing syrup according to claim 1 or 2, wherein the viscosity is 20,000 cP or less.   The plant-derived β-glucan-containing syrup according to any one of claims 1 to 3, wherein Brix is 30% to 80%.   The plant-derived β-glucan-containing syrup according to any one of claims 1 to 4, wherein the pH is 4.0 to 8.0.   The plant-derived β-glucan-containing syrup according to any one of claims 1 to 5, wherein the plant-derived β-glucan is a gramineous plant-derived β-glucan.   The plant-derived β-glucan-containing syrup according to any one of claims 1 to 6, which is obtained by a production method including a reaction step using glucoamylase.   A food or beverage comprising the plant-derived β-glucan-containing syrup according to any one of claims 1 to 7.

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