JP6837584B2 - Plant-derived β-glucan-containing syrup - Google Patents

Description

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

β-Glucan has glucose as a constituent unit, and these are β-1,3 bonds, β-1,
It is a linear polymer that binds four. It has been reported that this β-glucan has physiological functions such as normalization of blood cholesterol, suppression of increase in blood glucose level after meals, and sustained action of satiety.

In particular, the following mechanisms are known in terms of normalizing blood cholesterol. Cholesterol is absorbed into the small intestine by associating with micelles of bile acid and incorporating cholesterol into micelles of bile acid. β-Glucan inhibits the formation of these micelles or destroys the formed micelles to prevent cholesterol from being absorbed into the small intestine.

However, such an inhibitory ability has been confirmed only in 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 particular such effect has been reported with molecular weight β-glucan.

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

However, for example, when considering the use of 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 it is difficult to handle. In addition, Patent Document 1 also describes that a syrup having a sufficiently low viscosity was obtained, but it was achieved by simply reducing the Brix of the obtained syrup to less than 10%. As a syrup for foods and beverages, there is a problem in handling it from the viewpoint that the β-glucan content is low and the risk of microbial growth during syrup storage is high because of its low Brix. 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 having an ability to inhibit cholesterol micelle formation and having excellent functionality. Another object of the present invention is to provide new foods and beverages to which the plant-derived β-glucan-containing syrup is added.

In order to solve the above problems, the present inventors have a weight average molecular weight of 2,500 to 40,000.
A plant-derived β-glucan-containing syrup containing 0 plant-derived β-glucan was produced, and it was found that such a plant-derived 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 having a weight average molecular weight of 2,500 to 40,000 is contained in a content of 2% by mass to 8% by mass with respect to the total soluble solid content. Contains 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 formation into micelles.

A preferred embodiment of the present invention is
(3) The plant-derived β-glucan-containing syrup according to (1) or (2) above, which has a viscosity of 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, characterized in that 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, which has a pH of 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 grass-derived β-glucan.

A preferred embodiment of the present invention is
(7) The 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.

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

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide a plant-derived β-glucan-containing syrup having a sufficiently low viscosity and being easy to handle. In addition, it is possible to provide a plant-derived β-glucan-containing syrup having an ability to inhibit cholesterol micelle formation and having excellent functionality despite being easy to handle. Furthermore, it is possible to provide new foods and beverages to which the plant-derived β-glucan-containing syrup is added.

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 the viscosity in the plant-derived β-glucan-containing syrup of Example 1. FIG. 4 is a diagram showing the results of an evaluation test of the ability of each syrup ingested in subject A to suppress an increase in blood glucose level. FIG. 5 is a diagram showing the results of an evaluation test of the ability of each syrup ingested in subject B to suppress an increase in blood glucose level.

In the present invention, a plant-derived β-glucan-containing syrup containing 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 based on the total soluble solid content is produced. doing. No such plant-derived β-glucan-containing syrup is suggested in the prior art documents. Specifically, the present invention can be implemented as follows.

<Plant-derived β-glucan>
As an example, a plant containing β-glucan is used as a raw material for the syrup according to the present invention. The forms of such "plants" containing β-glucan include leaves, petals, stems, and so on.
It may be any of xylem, root, epidermis, fiber cell, branch, fruit, seed and the like.

Preferred examples of the "plant" used as a raw material include gramineous plants such as rice, wheat, corn, sorghum, barley, millet, millet, barley, oats, and rye. Preferred are barley and oats, and more preferably barley. The raw material may be selected from these plants according to the intended use and purpose, and may be used alone or in combination as appropriate.

When barley is used as a raw material, any variety of barley such as two-row barley, four-row barley, six-row barley, and bare barley can be used. In addition, as its form, whole grains, refined wheat grains,
Any tissue or fraction of barley seeds such as bran can be used. In barley seeds, since the endosperm contains a large amount of β-glucan, it is more preferable to use whole grains containing endosperm and milled wheat grains, and it is particularly preferable to use milled wheat grains.

<Manufacturing method of plant-derived β-glucan-containing syrup>
Hereinafter, the method for producing a plant-derived β-glucan-containing syrup according to the present invention will be described in the case where barley is used as a raw material, but the raw material used is not limited to barley, and naturally the above plants can be used in addition to barley. Is. Further, this production method is also an example of a method for producing a plant-derived β-glucan-containing syrup, and of course, another production method can be used.

1. 1. Preparation Step First, in producing the syrup according to the present invention, the crushed barley seeds are mixed with pure water with appropriate stirring. Then, a solution mixed with crushed barley seeds is placed in a constant temperature bath at 30 ° C to 6 ° C.
Stir while maintaining at 0 ° C. to completely disperse the crushed barley in pure water.

The concentration of the crushed barley seeds at this time is preferably 5% to 50%, more preferably 10% to 30% on a mass basis (W / W) with respect to the total amount of the solution. If this concentration is higher than 50%, a sufficient liquefaction reaction cannot be carried out in the liquefaction step.
Undecomposed starch remains, which causes a decrease in commercial value. In addition, the solution in which undecomposed starch remains causes an increase in viscosity, which makes solid-liquid separation in the filtration step and the filter treatment step difficult.

Further, the pH of the solution is preferably adjusted to 3.0 to 8.0, more preferably pH 4.0 to 7.0, and further preferably pH 5.0 to 6.0 by using an appropriate pH adjusting agent. Is desirable. When the pH is 8 or less, coloring due to the reaction is suppressed, and the flavor of the obtained syrup is improved.
When the pH is 3 or more, the enzyme activity is high and the reaction can be sufficiently carried out. The pH adjuster used may be any one suitable for foods or beverages. As an example, as a pH adjuster used to lower the pH, 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, still more preferably malic acid. Also, p used to raise the pH
As the H adjusting agent, 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. However, more preferably calcium hydroxide or sodium hydroxide is used, and even more preferably calcium hydroxide is used.

Then, the proteolytic enzyme is added to the solution in which the pulverized barley is dispersed, the temperature of the solution is raised to the reaction temperature of the added proteolytic enzyme, and the reaction is carried out in a predetermined reaction time.
It is important that the proteolytic reaction is performed before the liquefaction reaction. Since proteins are present around β-glucan, the enzyme cannot approach β-glucan without a proteolytic reaction. Therefore, in the end, only a syrup that does not contain enough β-glucan can be obtained.

As the proteolytic enzyme to be added, any one having an EC number of the 3.4 group may be used, and a protease (EC 3.4.22.2) can be preferably used. As such a proteolytic enzyme, a commercially available product may be used, and the trade name "Sumiteam P" (trade name "Sumiteam P") is particularly preferable.
(Made by Shin Nihon Kagaku Kogyo Co., Ltd.).

The amount of proteolytic enzyme added is preferably 0.0 on a mass basis (W / W) with respect to the raw material.
01% to 2.0%, more preferably 0.01% to 1.0%, even more preferably 0.03% to
It is 0.1%. If it is 2.0% or less, the 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 also differ depending on the type of proteolytic enzyme to be added and the plant used as a raw material. However, as an example, preferably 30 ° C. to 65 ° C.
Reaction temperature of ° C., more preferably reaction temperature of 40 ° C. to 65 ° C., still more preferably 45 ° C. to 60.
The reaction temperature is ° C., preferably 0.05 to 24 hours of reaction time, more preferably 0.2 to 12 hours of reaction time, still more preferably 0.3 to 6 hours of reaction time, in particular. The reaction time is preferably 0.5 hour to 2 hours.

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

As the liquefying enzyme to be added, any enzyme that randomly cleaves the α-1,4 glycosidic bond of starch may be used, and α-amylase (EC 3.2.1.1) can be preferably used. As such a liquefiing enzyme, a commercially available product may be used, and a particularly preferable product is the trade name "Crystase T10S" (manufactured by Amano Enzyme Co., Ltd.).

The amount of liquefied enzyme added should not be poorly liquefied, but it is based on the mass of the raw material (W / W).
) Is preferably 0.001% to 1.0%, more preferably 0.03% to 0.5%, and further preferably 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 liquefiing enzyme differ depending on the type of the liquefiing 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., and the reaction time is preferably 0.01 to 24 hours, more preferably 0.1. The reaction time is from time to 12 hours, more preferably from 0.1 hour to 2 hours.

3. 3. Saccharification step After lowering the liquefied enzyme solution to the reaction temperature of the saccharifying enzyme, the saccharifying enzyme and β-
Glucan-degrading enzyme is added, and a 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 inactivate the saccharifying enzyme and β-glucan degrading enzyme.

The saccharifying enzyme to be added may be any saccharifying enzyme that further decomposes dextrins generated by liquefaction into oligosaccharides, and a commercially available saccharifying enzyme can be used as appropriate.

When β-amylase (EC 3.2.1.2) is used as the saccharifying 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. The amount of such β-amylase added is preferably 0.001% to 2.0%, more preferably 0.1% to 1.5%, still more preferably, based on the mass of the raw material (W / W). Is 0.2
% To 1.0%.

Further, 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, preferably pullulanase (EC 3.21.41) or isoamylase (EC 3.2.1.68). ), Particularly preferably, the trade name "Pullulanase Amano 3" (manufactured by Amano Enzyme Co., Ltd.) can be used. The amount of such debranching enzyme added is preferably 0.001% to 2.0%, more preferably 0.
It is 01% to 1.5%, more preferably 0.01% to 1.0%.

Further, in the present invention, considering the use in foods and beverages, it is desirable that a low-viscosity solution is finally obtained. Therefore, it is desirable that the sugar composition contained in the finally obtained solution contains a larger amount of monosaccharides, disaccharides, and the like. Therefore, in addition to β-amylase and debranching enzyme, glucoamylase (EC 3.2.1.3) is further added as a saccharifying enzyme, and particularly preferably, the trade name “Dyzyme GPS” (manufactured by Amano Enzyme) is added. Can be done. The amount of this glucoamylase added is preferably 0.001% to 5.0%, more preferably 0.1% to 3.0%, and even more preferably 0. 2%
~ 2.5%.

Further, considering that the present invention aims to obtain a β-glucan-containing solution having a lower molecular weight, in the saccharification step, in addition to the above-mentioned 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 product can be used as appropriate, but it is particularly preferable to use the trade name “Fizym”.
"Registered trademark) 250L" (manufactured by Novozymes) can be used.

In the crushed barley used in this production method, β- of several millions in weight average molecular weight is generally used.
Contains glucan. Therefore, from the viewpoint of obtaining the viscosity of the final solution and the desired ability to inhibit micelle formation, β- with a desired weight average molecular weight was decomposed by β-glucan degrading enzyme.
You get glucan. By adjusting the addition amount of the β-glucan degrading enzyme and the pH at the time of reaction, it is possible to control the weight average molecular weight of β-glucan within a predetermined range.

In the present invention, the preferable weight average molecular weight of β-glucan is preferably 2,50.
0 to 40,000, more preferably 6,000 to 30,000, even more preferably 7,000
It is 0 to 25,000, particularly preferably 8,000 to 20,000. The amount of β-glucan degrading enzyme used to obtain this weight average molecular weight is preferably 0.0001% to 1.0%, more preferably 0., based on the mass of the raw material (W / W). 001% ~
It is 0.1%, more preferably 0.002% to 0.01%. The pH of the solution during the decomposition reaction by the β-glucan degrading enzyme is not particularly problematic as long as the enzyme is not inactivated in a short period of time, but is preferably pH 4.0 to pH 8.0, more preferably pH 4. 5-pH 7.0
More preferably, the pH is 5.0 to 6.0.

The reaction temperature and reaction time in this step differ 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, preferably 40 ° C.
Reaction temperature of ~ 80 ° C, more preferably reaction temperature of 50 ° C to 70 ° C, still more preferably 55 ° C.
The reaction temperature is ~ 65 ° C., preferably 0.1 hour to 24 hours reaction time, more preferably 0.5 hour to 12 hours reaction time, still more preferably 1 hour to 2 hours reaction time. Be done.

4. Filtration / Filtering / 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 the insoluble portion has been removed. And Br according to the use
Concentration is performed using an evaporator or the like so as to be ix.

A plant-derived β-glucan-containing syrup can be obtained by the above preparation step, liquefaction step, saccharification step, filtration / filtering / concentration step.

<Brix (soluble solid content concentration)>
In the present invention, Brix is a soluble solid content concentration (%), and the refractive index of an aqueous solution in which the soluble solid content is dissolved is measured at 20 ° C., and ICUMSA (Internati) is used.
onal Commission for Uniform Methods of S
It is a value converted into mass / mass percent of a pure sucrose solution based on a conversion table provided by ugar Analysis). In the present invention, it means the concentration of the soluble solid content contained in the aqueous solution, that is, in the plant-derived β-glucan-containing syrup produced in the above production step. This Brix can be measured by appropriately using a known measuring method that is already known, and is generally a commercially available sugar content meter (for example, a digital refractometer trade name "RX-5000α").
": Can be measured using Atago Co., Ltd.).

The optimum 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 produced syrup.

<Ability to inhibit micelle formation of β-glucan>
Bile acids involved in the absorption of cholesterol form bile acid micelles above a predetermined concentration. Cholesterol is not absorbed by the small intestine alone, but can be absorbed from the small intestine by being taken up by micelles of bile acids. That is, cholesterol is absorbed into the small intestine as micelles taken up by bile acid micelles.

At this time, β-glucan inhibits the association of cholesterol with micelles of bile acid, or destroys the micelles once formed and releases cholesterol again, thereby absorbing cholesterol in the small intestine. Is inhibited.

That is, in a broad sense, the ability of cholesterol to inhibit micelle formation means that cholesterol is taken up by bile acid micelles to form new micelles, thereby inhibiting absorption into the small intestine.

Then, it can be confirmed by the following test whether or not the plant-derived β-glucan-containing syrup obtained by the above production method has the ability to inhibit cholesterol micelle formation.

First, in addition to bile acid salts and cholesterol, other lipids that can be absorbed in the body (eg,
Prepare a micelle solution containing lecithin, oleic acid, etc.). 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 first prepared micelle solution, and the mixture is incubated at room temperature to 40 ° C. for 12 hours to 48 hours. Then, the solution after incubation 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.

For the extraction of β-glucan in the plant-derived β-glucan-containing syrup, a known method can be appropriately used as described above, and as an example, AOAC (Association).
of Official Analytical Chemists) Intern
It is preferable to use the method specified in AOAC 985.12, which is the official method of national. Further, a commercially available kit using this method can be used, for example, the product name "T".
"Otal Dietary Fiber Assay" (manufactured by Sigma-Aldrich) can be mentioned as a commercially available kit that can be used.

In addition, whether or not the β-glucan solution extracted by the above method has the ability to inhibit micelle formation can be qualitatively determined using a standard product. For example, when a barley-derived β-glucan-containing syrup is used in the test, a β-glucan solution derived from the obtained barley β-glucan preparation is prepared by decomposing it with a β-glucan degrading enzyme. Then, after adding a predetermined amount of the β-glucan solution derived from the standard to the micelle solution prepared first, the concentration of free cholesterol is measured through incubation, centrifugation and the like in the same manner as in the above test. The barley β-glucan preparation thus obtained and the β-glucan derived from the above production method are used to carry out the above test, and the free cholesterol concentration is compared to evaluate the ability to inhibit micelle formation.

<Ability to suppress blood sugar elevation>
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 sufficiently hungry healthy person is allowed to ingest a predetermined amount of the plant-derived β-glucan-containing syrup according to the present invention as the first meal (first meal), and then the blood glucose level is measured at regular intervals. It is possible to confirm by.

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 increase in blood glucose level by the next meal (second meal) was determined at regular intervals. Evaluate to. This makes it 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 preferably 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 good to contain β-glucan. This not only facilitates handling as a raw material for foods and beverages, but also enables the physiological function of β-glucan to be fully exerted.

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 or more.
It is preferably 15,000 cP, more preferably 2,000 cP to 10,000 cP. The Brix of the plant-derived β-glucan-containing syrup is preferably 30.
It is desirable that it is% to 80%, more preferably 50% to 80%, and even 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 even more preferably pH.
It is desirable that the pH is 5.0 to 6.0.

<Foods or beverages 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 are fruits (cut fruits such as apples, bananas and melons, pickled syrup, etc.) and processed cereals (
Bread, rice cakes, etc.), processed meat products (ham, sausage, etc.), dairy products (butter, cheese, yogurt, etc.), processed fruit products (jam, marmalade, etc.), confectionery (chocolate, cookies, cakes, jellies, etc.), Various foods such as seasonings (soy sauce, sauces, marmalade, etc.) and supplements can be mentioned. Examples of beverages include beer, non-alcoholic beverages,
Various beverages such as juice, coffee, black tea, green tea and carbonated beverages can be mentioned.

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, lactose, sorbitol, erythritol, maltitol,
Sugar alcohols such as xylitol, high-sweetness sweeteners such as aspartame, stebioside, sclarose, and acesulfam K, organic acids such as citric acid, tartrate acid, malic acid, succinic acid, and lactic acid, L-ascorbic acid, dl-α-tocopherol. , Vitamin Bs, nicotinic acid amide, vitamins such as calcium pantothenate, glycerin fatty acid ester, polyglycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, propylene glycol fatty acid ester and other surfactants, Arabic gum, carrageenan, Thickeners such as pectin and agar, stabilizers such as casein and gelatin, minerals such as amino acids and calcium salts, additives such as sodium erythorbic acid, glycerin and propylene glycol, foods or beverages such as pigments, fragrances and preservatives. The material can be mentioned.

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

Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited thereto. In the following examples, unless otherwise specified, "%" means mass / mass%.

[Example 1]
In Example 1, the following materials and methods were used.
<Preparation of plant-derived β-glucan-containing syrup>
After dispersing the crushed barley with pure water so as to be 30%, the mixture was stirred while heating to 50 ° C. in a constant temperature bath until the lumps of the crushed barley disappeared and the barley was completely dispersed in pure water. Then, a 10% calcium hydroxide solution was added to adjust the pH of the pulverized barley solution to 5.6. Next, the trade name "Sumiteam P" (manufactured by Shin Nihon Kagaku Kogyo Co., Ltd.) was added to the barley crushed product solution as a proteolytic enzyme (EC 3.4.22.2) at 0.05% based on the mass of the raw material (W / W). , The trade name "Crystase T10S" as α-amylase (EC 3.2.1.1), which is a liquefied enzyme.
(Manufactured by Amano Enzyme) was added at a concentration of 0.4% based on the mass of the raw material (W / W).
The reaction was carried out at 50 ° C. for 1 hour.

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

Next, the reaction solution was cooled to 60 ° C. in a constant temperature bath, and β-amylase (E) was used as a saccharifying enzyme.
Pullulanase (EC3) with the trade name "β-amylase # 1500S" (manufactured by Nagase ChemteX Corporation), which is C3.2.1.2), at a concentration of 0.52% based on the mass of the raw material (W / W).
.. 2.1.41) The trade name "Pullulanase Amano 3" (manufactured by Amano Enzyme) was used as a glucoamylase (EC 3.2.1.) At a concentration of 0.26% based on the mass of the raw material (W / W).
.. 3) The product name "Dyzyme GPS" (manufactured by Amano Enzymes) was used at a concentration of 0.26% based on the mass of the raw material (W / W) with β-glucan degrading enzyme (EC 3.21.73). A certain trade name "Finizem (registered trademark) 250L" (manufactured by Novozymes) was added to the above reaction solution at a concentration of 0.0052% based on the mass of the raw material (W / W). Then, a saccharification reaction was carried out at 60 ° C. for 2 hours, and then the mixture was heated to 90 ° C. to inactivate the enzyme.

Next, a filter paper (trade name "qualitative filter paper No. 2": manufactured by ADVANTEC) covered with diatomaceous earth (trade name "Radiolite # 500S": manufactured by Showa Chemical Industry Co., Ltd.) was put on a nutche at 80 ° C.
The above reaction solution kept at ~ 85 ° C. was passed through. This filtrate is used as a membrane filter with a pore size of 0.8 μm (trade name “Mixed Cellulose Esters (MCE)).
Membrane Filters ": made by ADVANTEC)
The above-mentioned filtrate kept at 70 ° C. to 75 ° C. was passed through. The obtained filtrate was concentrated by an evaporator to the extent that it did not stick. The term "sticking" here means a state in which a part of the syrup on the bottom surface of the flask loses its fluidity and adheres to the flask during concentration.

<Method for quantifying β-glucan>
The quantification of β-glucan in the plant-derived β-glucan-containing syrup prepared by the above method is performed by the β-glucan measurement kit (trade name “MIXED LINKE BETA-GLU”).
Using the CAN Assay KIT (manufactured by Megazyme), the McCreery method (manufactured by Megazyme)
The enzyme method) was used. That is, 5 ml of a plant-derived β-glucan-containing syrup adjusted to Brix 10% was placed in a 15 ml tube, and 2.5 g of finely crushed ammonium sulfate was added and dissolved. After allowing to stand at 4 ° C. for 20 hours, the supernatant was removed by centrifugation at 4 ° C. and 3,000 rpm for 10 minutes. Add 1 mL of a 50% (V / V) ethanol aqueous solution to the remaining pellets.
Stir vigorously to suspend the pellet, then add 10 mL of 50% (V / V) ethanol aqueous solution.
In addition, it was mixed. The supernatant was removed by centrifuging again at 4 ° C. and 3,000 rpm for 5 minutes. The operations of suspension of pellets, addition of ethanol, centrifugation and removal of supernatant were repeated again. 2 pellets
After redissolving in 4.8 mL of 0 mM sodium phosphate buffer (pH 6.5), 1 with the above buffer.
200 μL of the lichenase solution was added to 4.8 mL of the 0-fold diluted solution, and the mixture was incubated at 40 ° C. for 5 minutes. Centrifugal supernatant at 4 ° C., 3,000 rpm for 10 minutes, 100 μL each, 15 m
Transferred to l-tube. Add 100 μL of β-glucosidase solution to the tube and add at 40 ° C., 15
It was reacted for a minute. Then put the glucose oxidase / peroxide on the tube.
3 mL each of se (GOPOD) was added, and the mixture was reacted at 40 ° C. for 20 minutes, and the absorbance at 510 nm was measured. The absorbance of 100 μg of glucose was separately measured according to the method described in the kit and used as a reference. Instead of β-glucosidase solution, 50 mM acetate buffer (pH 4.
0) 100 μL was added as a blank. The β-glucan concentration was calculated by the following formula.

β-Glucan concentration (% with respect to soluble solids) = ΔA × F × 9 × D × (100 / B)
rix)
Here,
ΔA = sample absorbance-blank absorbance F = 100 / glucose 100 μg absorbance D = dilution factor when syrup is diluted

<Method of extracting β-glucan and method of 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, the measurement kit (trade name "Total Dietary Fiber Assay":
Purified using Sigma-Aldrich). That is, 1 g of plant-derived β-glucan-containing syrup and 50 mL of 0.08 M phosphate buffer (pH 6.0) were placed in an Erlenmeyer flask.
, Add 100 μL of the α-amylase (EC 3.2.1.1) solution included in the above kit.
After covering with aluminum foil, it was incubated in boiling water for 30 minutes. Then at room temperature 3
After cooling for 0 minutes, about 10 mL of a 0.275N sodium hydroxide aqueous solution was added to the above reaction solution to adjust the pH to 7.5 ± 0.2. Then, 100 μL of a solution obtained by diluting the protease (EC 3.4.21.62) solution included in the above kit to 50 mg / mL was added to the reaction solution, and the mixture was incubated at 60 ° C. for 30 minutes with shaking, and then 30 at room temperature. Cooling was performed for a minute. Next, 10 mL of a 0.325 N hydrochloric acid aqueous solution was added to the above reaction solution to adjust the pH.
It was adjusted to 3 ± 0.3. After that, the amyloglucosidase (EC3.2) included in the above kit
.. 1.3) 100 μL of the solution was added to the reaction solution and incubated at 60 ° C. for 30 minutes with shaking. Then, the reaction solution was boiled for 3 minutes to inactivate the above enzyme. Then, the reaction solution was passed through a 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 above reaction solution. , Solid layer extraction column (trade name "Sep-Pak C18 Cartridg"
e ”: Made by Waters Corp.). 5 mL of the passed reaction solution 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 at 4 ° C for 10 minutes
After centrifugation at 3,000 rpm, the supernatant was discarded, 5 mL of 75% ethanol was added to the pellet, and the pellet was washed with a vortex mixer. Then, again at 4 ° C. for 10 minutes, 3
After centrifugation at 000 rpm, the supernatant was discarded, 5 mL of 75% ethanol was added to the pellet, and the pellet was washed with a vortex mixer. After repeating the steps of centrifugation and washing again, pure water was added to the obtained pellets and dissolved by heating at 80 ° C. or higher. The solution is applied to a membrane filter having a pore size of 0.45 μm (trade name “MILLEX®”).
-HP 0.45 μm ”: manufactured by MILLIPORE). The β-glucan solution is
Its molecular weight distribution and weight average molecular weight were measured by gel filtration chromatography.

Measurement of molecular weight distribution and weight average molecular weight by gel filtration chromatography is performed by HLC-8.
A 220 GPC (manufactured by Tosoh Corporation) was used under the following conditions.
Column: G2500PW-G3000PW-G6000PW (manufactured by Tosoh Corporation) 3-link column Solvent: Pure water Temperature: 80 ° C
Flow velocity: 1.0 ml / min
Detection: RI (Differential Refractometer)
The obtained molecular weight distribution and weight average molecular weight are obtained by pullulan (trade name "STA") having a known molecular weight.
It was calculated based on a calibration curve prepared using "NDARD P-82": manufactured by Shodex).

<Viscosity measurement method>
Put 1.2 mL each of the plant-derived β-glucan-containing syrup obtained by the above production method into a plate of an E-type viscometer (trade name "VISCONIC ED": manufactured by Tokyo Keiki Co., Ltd.) to determine the viscosity of the syrup in each Brix. It was measured. The temperature at the time of measurement was 25 ° C.

<Results of characterization 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. Then, the characteristics of Brix of the syrup according to Example 1 and the concentration, molecular weight distribution, and weight average molecular weight of the extracted β-glucan were evaluated.

[Comparative Example 1]
As Comparative Example 1, a special barley flour (β-glucan content of 7.5% by mass) was used as a raw material, and the trade name “Dyzyme” was 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 a transglucosidase (EC 3.2.1.20), is used as a transfer enzyme based on the mass of the raw material (W / A plant-derived β-glucan-containing syrup obtained by the same method as the above production method was prepared except that it was added at 0.1% in W). Here, the β-glucan-containing syrup produced in Comparative Example 1 was used as Bri in the concentration step.
It stuck at x30% or less. Then, Brix of the syrup according to Comparative Example 1 and the extracted β
-Glucan concentration, molecular weight distribution, and weight average molecular weight were evaluated. The molecular weight distribution and the weight average molecular weight were measured by using a high performance liquid chromatograph Prominence (manufactured by Shimadzu Corporation) under the following conditions of gel filtration chromatography.
<Measurement conditions>
Column: GS520-GS320 (manufactured by Shodex)
Solvent: Pure water Temperature: 50 ° C
Flow velocity: 1.0 ml / min
Detection: CAD (Corona charged particle detector: manufactured by Nippon Dionex)

First, FIG. 1 shows β- from the plant-derived β-glucan-containing syrup obtained in Example 1.
It is a figure which shows the result of having extracted glucan and measuring the molecular weight distribution by gel filtration chromatography. FIG. 2 shows β from the plant-derived β-glucan-containing syrup obtained in Comparative Example 1.
-It is a figure which shows the result of having extracted glucan and measuring the molecular weight distribution by gel filtration chromatography. In addition, Table 1 is a calculation of the molecular weight of β-glucan and the amount of β-glucan 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 β-glucan having a molecular weight of 2,000 to 10,000 and β-glucan having a molecular weight of less than 2,000.
A large amount of glucan is present, and a large amount of lower molecular weight β-glucan can be obtained by the above production method.

Table 2 shows the results of comparing the characteristics of the weight average molecular weight, the β-glucan concentration, and the maximum Brix that can exist as a syrup between Example 1 and Comparative Example 1. Generally Br
As the ix is increased, the fluidity of the syrup is lost, and the syrup cannot be handled substantially. The Brix has a fluidity that can be handled as a syrup.
It indicates the maximum value of x. Specifically, the presence or absence of sticking 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. Do you get it. Further, it was found that the weight average molecular weight was 13,000, which was sufficiently lower than the weight average molecular weight of Comparative Example 1. In general, low Brix increases the risk of microbial growth and is unsuitable for use as food or beverage. But on the other hand, Br
Increasing the ix 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 shows a sufficiently low weight average molecular weight, and even if it has a high Brix of 70% or more, it is a syrup having an appropriate viscosity, and is used for foods and foods. Expected to be used as a beverage.

<Results of characterization of plant-derived β-glucan-containing syrup>
Regarding the syrup according to Example 1, in addition to the above characteristics, characteristics such as the composition ratio of monosaccharides, disaccharides, trisaccharides, saccharides of tetrasaccharides and above, and the viscosity of the syrup were also evaluated. The method for evaluating the sugar composition is as follows. The syrup was diluted to about 10% Brix, and activated carbon (trade name "Shirasagi A": manufactured by Japan Enviro Chemicals Co., Ltd.) was added in an appropriate amount and mixed. Next, the sample solution was heated until it was boiled and passed through a filter paper (trade name "qualitative filter paper No. 2": manufactured by ADVANTEC). Then, 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, and then solidified. Layer extraction column (trade name "Sep-Pak C18 Cartri"
"Dge": manufactured by Waters Corp.) was passed through the liquid. Adjust the Brix of the sample to about 1% to 5%, pass it through a membrane filter with a pore size of 0.45 μm (trade name “MILLEX®-HP 0.45 μm”: manufactured by Merck & Co., Inc.), and sugar. It was used as a sample for composition analysis. The sugar composition was analyzed using the trade name "Alliance (registered trademark) HPLC system" (manufactured by Japan Waters Corp.).
<Measurement conditions>
Column: ULTRON PS80-N (manufactured by Shimadzu LLC)
Solvent: Pure water Temperature: 60 ° C
Flow velocity: 0.6 ml / min
Detection: RI (Differential Refractometer)

[Example 2]
Further, as Example 2, the plant-derived β produced by the same method except that the glucoamylase to be added was set to 0.052% based on the mass standard (W / W) with respect to the raw material from the production method according to Example 1 above. -A syrup containing glucan was prepared. Then, with respect to the syrup according to Example 2, characteristics such as the composition ratio of monosaccharides, disaccharides, trisaccharides, and saccharides above tetrasaccharides and the viscosity of the syrup were evaluated in the same manner as in Example 1.

[Example 3]
In addition, as Example 3, all plant-derived β produced by the same method except that the glucoamylase to be added was 2.08% based on the mass standard (W / W) with respect to the raw material from the production method according to Example 1 above. -A syrup containing glucan was prepared. Then, with respect to the syrup according to Example 2, characteristics such as the composition ratio of monosaccharides, disaccharides, trisaccharides, and saccharides above tetrasaccharides and the viscosity of the syrup were evaluated in the same manner as in Example 1.

[Example 4]
In addition, as Example 4, a plant-derived β-glucan-containing syrup produced by the same method except that glucoamylase was not added was prepared from the production method according to Example 1 above. Then, with respect to the syrup according to Example 2, characteristics such as the composition ratio of monosaccharides, disaccharides, trisaccharides, and saccharides above tetrasaccharides and the viscosity of the syrup were evaluated in the same manner as in Example 1.

Table 3 shows the results of comparing the characteristics of Examples 1 to 4 such as the composition ratio of monosaccharides, disaccharides, trisaccharides, and saccharides above tetrasaccharides and the viscosity of syrup.

As is clear from Table 3 above, all of the plant-derived β-glucan-containing syrups according to Examples 1 to 4 have a weight average molecular weight in the range of 2,500 to 40,000 β-.
Glucan 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 was 3.8% to 4.2% with respect to the soluble solid content. Had a β-glucan concentration of.

Also, in general, syrup is desired to have a Brix of 70% or more, for example, from the viewpoint of reducing the risk of microbial growth during storage, but on the other hand, such a high Brix.
The syrup has a high viscosity and is not suitable for use as food or beverage.

However, 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 all have an appropriate viscosity as the viscosity when Brix is 70% or more. Was found to have. In particular, since the plant-derived β-glucan-containing syrups obtained in Examples 1 to 3 were added with glucoamylase in the saccharification step, the plant-derived β-glucan-containing syrup obtained in Example 4 to which it was not added was added. -Compared to glucan-containing syrup, the proportion of monosaccharides in the sugar composition is large. Therefore, the viscosity of the plant-derived β-glucan-containing syrups of Examples 1 to 3 is as low as 3,100 cP to 9,900 cP, as opposed to the plant-derived β-glucan-containing syrup of Example 4. It was found that it has very excellent properties as a raw material for foods and beverages.

Further, FIG. 3 is a diagram showing the results of measuring the relationship between each Brix and the viscosity in the plant-derived β-glucan-containing syrup of Example 1. According to this, the Brix described above is 7
Even if it is 0% or more, it has an appropriate viscosity of 10,000 cP or less.

Further, for example, when considering application as a raw material for foods and beverages, naturally, the plant-derived β-glucan-containing syrups of Examples 2 to 4 are also sufficiently utilized as raw materials for foods and beverages. Although it is possible, 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 in which a larger amount of glucoamylase was added in the saccharification step contains a large amount of monosaccharides, it is considered that saccharides may be precipitated when Brix is increased. In addition, Example 2 in which a smaller amount of glucoamylase was added in the saccharification step.
And, the plant-derived β-glucan-containing syrup of Example 4 in which no addition was made was carried out because it contained a large amount of saccharides of disaccharide or more with respect to the plant-derived β-glucan-containing syrup of Example 1. Compared with the plant-derived β-glucan-containing syrup of Example 1, its viscosity is slightly higher.

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

<Evaluation method of micellar inhibitory ability>
The ability of plant-derived β-glucan-containing syrups obtained by the above production method to inhibit cholesterol formation was evaluated by Nagaoka et al. , The Journal
of Nutrition (1999), vol. 129, pp. 1725-173
The method described in 0 was appropriately modified. Specifically, it was carried out through each of the following steps.
1. 1. Preparation of Micelle Solution The micelle solution used to evaluate the ability of the plant-derived β-glucan-containing syrup obtained by the above production method to inhibit cholesterol formation was prepared as follows.

The micelle solution was prepared in a glass bottle by adding 250 mL of 15 mM phosphate buffer (pH 7.4) to each reagent shown in Table 4 to a predetermined final concentration. Then, a glass bottle containing the solution was placed in an ultrasonic cleaner (trade name "UT-604": manufactured by Sharp Corporation), and the above reagents were completely dispersed by ultrasonic waves for 30 minutes.

2. 2. Method for Extracting β-Glucan from Plant-Derived β-Glucan-Containing Syrup From the plant-derived β-glucan-containing syrup prepared in Example 1, the above-mentioned "Method for extracting β-glucan and method for measuring its molecular weight" is described. Β-Glucan was extracted using the method. The β-glucan pellets after ammonium sulfate precipitation and ethanol washing were stored at −80 ° C. for about 2 hours and frozen. Then, the pellets were freeze-dried for 20 hours or more at a vacuum degree of 0.133 mBar or less by a freeze-dryer (trade name "FreeZone (registered trademark)": manufactured by Love Conco).

3. 3. Preparation of β-glucan solution derived from standard Barley-derived β-glucan standard (manufactured by Sigma-Aldrich) is dissolved in pure water and 1
A plurality of% solutions were prepared. Next, lichenase (EC 3.21.73: Megazyme)
(Manufactured by the company) is added to each of 0.125 and 1.0 U solutions, the reaction is carried out at 38 ° C. for a predetermined time for each solution, and immediately after that time, the enzyme is heated at 90 ° C. for 10 minutes. Was deactivated. The amount of enzyme and reaction time used in each example are shown in Table 5. The molecular weight distribution of each of the obtained reaction solutions was measured by gel filtration chromatography. The measurement is "β-
The method was the same as that described in the section "Method for extracting glucan and method for measuring its molecular weight".

4. Evaluation test of micellar inhibitory ability β-glucan freeze-dried powder extracted from plant-derived β-glucan-containing syrup 2.5 mg
0.25 g of pure water and 0.25 g of 1% β-glucan solution obtained by enzymatically decomposing β-glucan preparation by the above method were placed in a 15 mL tube, and 4.75 g of micelle solution was added to β. -The final glucan concentration was 0.05% (W / W). At this time, as a positive control, 2.5 mg of a barley-derived β-glucan standard (manufactured by Sigma-Aldrich), whose cholesterol absorption inhibitory ability has already been confirmed, was suspended in 0.25 g of pure water as a micelle solution. Mixed. As a negative control, glucose 2.5 mg is pure water 0
.. What was suspended in 25 g was mixed with a micelle solution (Comparative Example 2). Further, 0.25 g of pure water was mixed with the micelle solution as a blank (Comparative Example 3).

Then, 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. After that, each reaction solution is applied to a swing rotor (trade name "SW28": manufactured by Beckman Coulter),
20:00 using a centrifuge (trade name "L7-55": manufactured by Beckman Coulter)
Centrifugation was performed at 0 rpm (100,000 g) for 60 minutes. 200 μL of the upper layer of each reaction solution was separated, 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 separated solution. Incubate the added solution at 37 ° C. for 5 minutes, and then use a spectrophotometer (trade name "BioSpec-mini": manufactured by Shimadzu Corporation) to make 6
The free cholesterol concentration was calculated by measuring the absorbance at 00 nm.

[Example 5]
As Example 5, β-extracted from the plant-derived β-glucan-containing syrup by the method described in the above section “2. Extraction method of β-glucan from the plant-derived β-glucan-containing syrup”.
Using glucan, the micellar inhibitory ability was evaluated by the method described in the section "4. Evaluation test of micellar inhibitory 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 standard", β-glucan standard derived from barley was added to the amount of lichenase added and the reaction shown in Table 5. By reacting with lichenase over time, β-glucan having the same weight average molecular weight as shown in Table 5 was prepared. Then, using this β-glucan, the micellar inhibitory ability was evaluated by the method described in the section "4. Evaluation test of micellar inhibitory ability". Similarly, as Examples 7 to 11, each β-glucan having a weight average molecular weight also shown in Table 5 was prepared at the amount of lichenase added and the reaction time shown in Table 5, and micelles were formed in each β-glucan. The inhibitory ability was evaluated.

[Reference Example 1] to [Reference Example 3]
As a reference example 1, according to the method described in the above section "3. Preparation of β-glucan solution derived from standard", β-glucan standard derived from barley was added to the lichenase addition amount and reaction time shown in Table 5. By reacting with lichenase, β-glucan having a larger weight average molecular weight than the 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 micelle formation was evaluated. Similarly, as Reference Example 2 and Reference Example 3, each β-glucan having a weight average molecular weight also shown in Table 5 was prepared at the amount of lichenase added and the reaction time shown in Table 5, and each β-glucan was prepared. The ability to inhibit micelles was evaluated.

[Comparative Example 2] and [Comparative Example 3]
Further, as Comparative Example 2, as described in the section "4. Evaluation test of micellar inhibitory ability",
Instead of β-glucan, 2.5 mg of glucose suspended in 0.25 g of pure water was used for the evaluation test of micellar inhibitory ability. Further, as Comparative Example 3, instead of β-glucan, pure water 0.
25 g was used for the evaluation test of micellar inhibitory ability.

Table 5 shows Examples 5 to 11, and Reference Examples 1 to 3, Comparative Example 2, and Comparative Example 3.
The results of evaluating each micelle formation inhibitory ability in the above based on the free cholesterol concentration (mg / dL) are shown.

According to Table 5, the β-glucan extracted from the plant-derived β-glucan-containing syrup of Example 5, which has a smaller weight average molecular weight (13,000), was also used as a negative control in Comparative Examples 2 and 3. Higher free cholesterol concentration (5)
9.4 mg / dL) was shown. Similarly, the weight average molecular weight of β-glucan according to the present invention is
The value is preferably 2,500 to 40,000, but the β-glucan showing the weight average molecular weight of Examples 6 to 11 is also higher than that of Comparative Example 2 and Comparative Example 3 used as the negative control. The free cholesterol concentration was shown.

Further, it has been conventionally reported that β-glucan having a very large weight average molecular weight (for example, Reference Example 3) has an ability to inhibit micelle formation. This is also consistent with the high free cholesterol concentration measured in Reference Example 3. On the other hand, in Examples 5 to 11,
The free cholesterol concentration was equal to or higher than that of Reference Example 1 and Reference Example 2 in which the weight average molecular weight was larger than that of β-glucan in Examples 5 to 11.

That is, even with β-glucan having a weight average molecular weight of 2,500 to 40,000 as in the present invention, a very high weight average molecular weight (300,00) as previously reported has been reported.
It was suggested that cholesterol can be prevented from being taken up by bile acid micelles to form new micelles and absorbed into the small intestine, similar to β-glucan which is (0 or more).

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

2. 2. Preparation of syrup containing no plant-derived β-glucan In the preparation of syrup containing no plant-derived β-glucan, a solution of crushed barley as a raw material is dispersed in pure water so that the crushed barley is 20%. The prepared point and the trade name "Finzym (registered trademark) 250" which is β-glucan degrading enzyme (EC 3.21.73).
Except for the fact that the amount of "L" (manufactured by Novozymes) was added to the reaction solution in an amount 10 times the amount of the preparation method of Example 1 above (concentration of 0.052% based on the mass of the raw material (W / W)). Was all prepared according to the preparation method of Example 1 above. Finally, a syrup containing no plant-derived β-glucan (0.2% by mass based on the total soluble solid content) was obtained.

3. 3. Preparation of syrup containing plant-derived β-glucan (weight average molecular weight: 81,000) In preparation of plant-derived β-glucan (weight average molecular weight: 81,000) -containing syrup, a crushed barley solution as a raw material is used as a crushed barley product. It was prepared by dispersing it in pure water so that the content was 20%, and the trade name "Fini" was β-glucan degrading enzyme (EC 3.21.73).
The reaction solution was added in an amount of "zym (registered trademark) 250 L" (manufactured by Novozymes) that was 1/2 times the amount of the preparation method of Example 1 above (concentration of 0.0026% based on mass (W / W) with respect to the raw material). Everything except the points added therein was prepared according to the preparation method of Example 1 above. Finally, a plant-derived β-glucan-containing syrup (weight average molecular weight: 81,000, 3.9% by mass based on the total soluble solid content) was obtained.

[Example 12] and [Example 13]
Two healthy subjects (both humans) were subjects (subject A: HbA1c NGSP value 5.1%,
Subject B: HbA1c NGSP value 5.1%), evaluation of the ability of the plant-derived β-glucan (weight average molecular weight: 12,000) -containing syrup prepared in 1 above to suppress the increase in blood glucose level according to the following test method. The test was performed. In addition, the result of the evaluation test by subject A is shown in Example 1.
The result of the evaluation test by Subject B and 2 was designated as Example 13.

First, each subject was allowed to refrain from eating or drinking other than water after 9:00 pm on the day before the test. Next, after measuring the blood glucose level in the early morning on an empty stomach, 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) with 50 g of sugar (the relevant). The sugar contained 2.1 g of β-glucan). Then, the blood glucose levels at 30, 60, 90, 120, and 150 minutes after the ingestion were measured, respectively. Furthermore, after measuring the blood glucose level on an empty stomach before lunch (240 minutes after ingesting the syrup), 150 g of cooked rice (52 g of sugar) was ingested as a second meal, and 270 minutes after ingesting the syrup.
Blood glucose levels were measured at 300 minutes, 330 minutes, and 360 minutes, respectively. In the above evaluation test
Blood glucose level is measured by enzyme method (hexokinase method) and automatic analyzer (manufactured by Beckman Coulter).
Was measured using. During the above evaluation test, the subjects were kept in a sitting position. The measurement results of blood glucose levels obtained from two healthy subjects by the above evaluation test are shown in Table 6 and FIG. 4, respectively.
And shown in FIG.

[Comparative Example 4] and [Comparative Example 5]
Using the same two healthy subjects as in Example 12 and Example 13 as subjects (subject A and subject B), 50 g of sugar (0.1 g of β-glucan in the sugar) was prepared for each subject in the syrup prepared in 2 above. Including), an evaluation test of the ability to suppress the rise in blood glucose level was conducted. The results of the evaluation test by subject A are shown in Comparative Example 4, and the results of the evaluation test by subject B are shown in Table 6, FIG. 4 and FIG. 5, respectively.

In the test methods of Comparative Examples 4 and 5, the syrups prepared in 2 above and which do not contain plant-derived β-glucan were used as the syrup to be ingested by each subject, except that the syrups of Examples 12 and 13 were used. It was carried out by the same method as the test method in. In addition, the above-mentioned Example 1
The evaluation test was conducted after a sufficient period (5 days or more) from the test date of 2 and Example 13.

[Comparative Example 6] and [Comparative Example 7]
Using the same two healthy subjects as in Example 12 and Example 13 as subjects (subject A and subject B), 50 g of sugar (2.0 g of β-glucan in the sugar) was prepared for each subject in the syrup prepared in 3 above. Including), an evaluation test of the ability to suppress the rise in blood glucose level was conducted. The results of the evaluation test by subject A are shown in Comparative Example 6 and the results of the evaluation test by subject B are shown in Table 6, FIG. 4 and FIG. 5, respectively.

The test methods in Comparative Examples 6 and 7 were described above, except that the plant-derived β-glucan (weight average molecular weight: 81,000) -containing syrup prepared in 3 above was used as the syrup to be ingested by each subject. It was carried out in the same manner as the test method in Example 12 and Example 13. In addition, the evaluation test was conducted after a sufficient period (5 days or more) from the test date of the above-mentioned Examples and Comparative Examples.

4 and 5 are diagrams showing the results of an evaluation test of the ability of each syrup ingested by subject A or B to suppress the increase in blood glucose level, respectively. In addition, Table 6 shows the specific blood glucose levels obtained in the evaluation test of the ability to suppress the increase in blood glucose level in each syrup ingested by Subject A and Subject B. According to this, in both subject A and subject B, plant-derived β-
When a syrup containing glucan (weight average molecular weight 12,000) was ingested as first meal on an empty stomach in the early morning (Examples 12 and 13), and when a syrup containing no plant-derived β-glucan was ingested as first meal. Compare (Comparative Example 4 and Comparative Example 5)
, The rise in blood glucose level was significantly suppressed. Furthermore, even after ingesting rice as the second meal, it is better to ingest the plant-derived β-glucan (weight average molecular weight 12,000) -containing syrup as the first meal (Examples 12 and 13). Compared with the case where the 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 maintained, and the so-called second meal effect was confirmed.

In Comparative Example 6 and Comparative Example 7 in which the plant-derived β-glucan (weight average molecular weight 81,000) -containing syrup was ingested, the plant-derived β-glucan (weight average molecular weight 81,000) was also ingested in the same manner as in Examples 12 and 13.
It was confirmed that the increase in blood glucose level was suppressed in Comparative Example 4 and Comparative Example 6 in which the syrup containing no glucan was ingested. However, the plant-derived β-glucan (weight average molecular weight 12,000) -containing syrup used in Examples 12 and 13 has a smaller weight average molecular weight than the plant-derived β-glucan (weight average molecular weight 81,000) -containing syrup. As confirmed in Examples 1 to 4 and Comparative Example 1, since it has a high Brix, the risk of microbial growth is low, and it has an appropriate viscosity, which is more desirable in food and beverage applications.

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide a plant-derived β-glucan-containing syrup having a sufficiently low viscosity, an ability to inhibit cholesterol micelle formation and an ability to suppress an increase in blood glucose level, and thus the present invention can be used in the food and beverage industries and the like. Is.

Claims (7)

A plant-derived β-glucan having a weight average molecular weight of 2,500 to 40,000 is contained in a content of 2% by mass to 8% by mass based on the total soluble solid content, the viscosity is 10,000 cP or less, and the Brix is 30. A plant-derived β-glucan-containing syrup characterized by being% to 80%. The plant-derived β-glucan-containing syrup according to claim 1, wherein the pH is 4.0 to 8.0. The plant-derived β-glucan-containing syrup according to claim 1 or 2, wherein the plant-derived β-glucan contained has an ability to inhibit cholesterol micellarization. The plant-derived β-glucan-containing syrup according to any one of claims 1 to 3, wherein the plant-derived β-glucan is a grass-derived β-glucan. The plant-derived β-glucan-containing syrup according to any one of claims 1 to 4, wherein the Brix is 50% to 80%. A plant-derived β-glucan-containing solution having a viscosity of 10,000 cP or less and a Brix of 30% to 80%, wherein the solution has a weight average molecular weight of 2,500 to 40,000. A plant-derived β-glucan-containing solution, which comprises a content of 2% by mass to 8% by mass with respect to the total soluble solid content. A food or beverage comprising the plant-derived β-glucan-containing syrup according to any one of claims 1 to 5 or the plant-derived β-glucan-containing solution according to claim 6.

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