JP2021106545A - Beer taste alcoholic beverage, agent for enhancing body of beer taste alcoholic beverage, and method for enhancing body of beer taste alcoholic beverage - Google Patents

Abstract

To provide a beer taste alcoholic beverage having strong body by using cereal-derived raw material.SOLUTION: A beer taste alcoholic beverage has a malt content of less than 50 wt.% in raw material, the beer taste alcoholic beverage having a high content of glycated lysine.SELECTED DRAWING: None

Description

The present invention relates to a beer-taste alcoholic beverage, a beer-taste alcoholic beverage richness enhancer, and a beer-taste alcoholic beverage richness-enhancing method.

With the diversification of consumer tastes in recent years, the development of beer-taste alcoholic beverages having various flavor characteristics is desired.

For example, Patent Document 1 describes an alcohol component in order to enhance the richness of a beer-like beverage.
It is disclosed that sweetness-based components, aldehyde-based components, etc. are used as flavor improving agents for beer-like beverages.

Japanese Unexamined Patent Publication No. 2016-214262

However, a flavor improving agent such as Patent Document 1 affects the flavor and may not be suitable for use.

In addition, grain-derived raw materials such as wheat and corn are commonly used as raw materials for beer-taste alcoholic beverages. Since the grain-derived raw material is a raw material commonly used in beer-taste alcoholic beverages, there is little resistance to the manufacturer and the beverage consumer to use it as a material for enhancing the richness of the grain-derived raw material. Therefore, a method of enhancing the richness by using a grain-derived raw material has been desired.

An object of the present invention is to provide a beer-taste alcoholic beverage having a strong rich taste by using a grain-derived raw material. Another object of the present invention is to provide a rich taste enhancer for beer-taste alcoholic beverages, which are raw materials derived from grains. Furthermore, an object of the present invention is to provide a method for enhancing the richness of a beer-taste alcoholic beverage using a grain-derived raw material.

That is, the present invention relates to the following beer-taste alcoholic beverages and the like.
[1] A beer-taste alcoholic beverage in which the proportion of malt in the raw material is less than 50% by weight, and contains a sugar-modified protein having a molecular weight of 35 to 50 kDa in which the amino group of lysine constituting a grain-derived protein is glycated. , A beer-taste alcoholic beverage having a glycated lysine content of 30.0 nmol / mL or more in the sugar-modified protein.
[2] The beer-taste alcoholic beverage according to the above [1], wherein the content of glycated lysine contained in the sugar-modified protein is 35.3 nmol / mL or more.
[3] The beer-taste alcoholic beverage according to the above [1] or [2], wherein the content of glycated lysine contained in the sugar-modified protein is 30.0 to 1000 nmol / mL.
[4] The beer-taste alcoholic beverage according to any one of the above [1] to [3], wherein the sugar glycated on the amino group of the glycated lysine is a monosaccharide or a disaccharide.
[5] The beer-taste alcoholic beverage according to any one of [1] to [4] above, wherein the grain is at least one selected from the group consisting of barley, wheat, corn, rice, and soybean.
[6] The beer-taste alcoholic beverage according to any one of the above [1] to [5], which does not contain wort.
[7] The rich taste of a beer-taste alcoholic beverage containing a sugar-modified protein having a molecular weight of 35 to 50 kDa and in which the amino group of lysine constituting a grain-derived protein is glycated, and the proportion of malt in the raw material is less than 50% by weight. Enhancer.
[8] To a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight, a sugar-modified protein having a molecular weight of 35 to 50 kDa to which the amino group of lysine constituting a grain-derived protein is ligated is added. A method for enhancing the richness of beer-taste alcoholic beverages, thereby enhancing the richness of beer-taste alcoholic beverages.

According to the present invention, it is possible to provide a beer-taste alcoholic beverage having a strong rich taste by using a grain-derived raw material. Further, it is possible to provide a rich taste enhancer for a beer-taste alcoholic beverage, which is a raw material derived from grains. Further, it is possible to provide a method for enhancing the richness of a beer-taste alcoholic beverage using a grain-derived raw material.

In the beer-taste alcoholic beverage of the present invention, the ratio of malt in the raw material is less than 50% by weight.
The malt ratio is preferably 30% by weight or more.
Moreover, the ratio of malt in the raw material may be 0%.
Here, the "malt ratio" refers to the ratio of the mass of malt to water and raw materials other than hops, such as malt, rice, corn, kouryan, potato, starch, wheat other than malt, and sugars. However, ingredients that can be added in trace amounts, such as acidulants, sweeteners, bitterness agents, seasonings, and flavors, are not included in the above ratio calculation.
The beer-taste alcoholic beverage of the present invention does not have to contain wort.

The beer-taste alcoholic beverage of the present invention contains a sugar-modified protein.
The sugar-modified protein has a molecular weight of 35 to 50 kDa.

Proteins with a molecular weight of 35 to 50 kDa are subjected to ultrafiltration using a 30 kDa cut-off membrane on the raw material solution of a beer-taste alcoholic beverage, and then a region having a molecular weight of 35 to 50 kDa measured by electrophoresis by SDS-PAGE. It is a protein found in.
A protein of about 40 kDa is preferable, and a protein of about 40 kDa is also referred to as a 40 kDa protein in the present specification.

The sugar-modified protein is a protein derived from grains.
The grain is preferably at least one selected from the group consisting of barley, wheat, corn, rice and soybean.
When the grain is wheat, it can contain a known wheat-derived protein used in the production of beer-taste alcoholic beverages. Examples of such wheat include barley, wheat, rye, oats, oats, and barley, and barley is preferable. Further, either germinated wheat or ungerminated wheat may be used, but germinated malt is preferable. These may be contained alone or in combination of two or more kinds.

Glycated proteins are proteins in which the amino group of lysine, which constitutes a grain-derived protein, is glycated.
"The amino group of lysine is glycated" in the present invention means that a sugar is bound to the amino group of lysine.
Of the amino groups of lysine, it is preferable that the sugar is glycated on the amino group of the side chain.
Confirmation that the amino group of lysine is glycated can be performed by colorimetric quantification by the NBT method (Nitroblue terrazolium method).
The sugar glycated on the amino group of lysine may be a monosaccharide, a disaccharide, or a polysaccharide of trisaccharide or more, but is preferably a monosaccharide or a disaccharide.
Monosaccharides include all sugars including glucose, fructose, galactose and mannose.
Examples of the disaccharide include maltose-type disaccharides such as maltose, lactose, and arabinose.
Examples of the trisaccharide include maltotriose, sertriose, fucosyl lactose, sialyl lactose and the like.

In the beer-taste alcoholic beverage of the present invention, the content of glycated lysine is 30.0 nmol / mL or more.
Further, the content of glycated lysine is preferably 35.3 nmol / mL or more, preferably 50.0 nmol / mL or more, preferably 80.0 nmol / mL or more, and preferably 100 nmol / mL. The above is also preferable.
Further, the content of glycated lysine is preferably 30.0 to 1000 nmol / mL, preferably 50.0 to 1000 nmol / mL, and preferably 80.0 to 1000 nmol / mL. It is also preferably 100 to 1000 nmol / mL.
The content of glycated lysine can be quantified by colorimetric quantification by the NBT method.

Since the beer-taste alcoholic beverage of the present invention contains the above-mentioned sugar-modified protein, it can be a beer-taste alcoholic beverage with a strong rich taste.
Further, since the sugar-modified protein is a raw material derived from grains, it is possible to prevent an unintended effect on the flavor unlike the flavor improving agent described in Patent Document 1. Further, since it is preferable that the raw materials for the beer-taste alcoholic beverage are only grains and hops, it is a preferable feature as the raw material for the beer-taste alcoholic beverage that the sugar-modified protein is a grain-derived raw material.

In the present invention, the "rich taste" is a taste that cannot be expressed by five basic tastes, that is, sweetness, saltiness, acidity, bitterness and umami, and the strength of taste (drinking response), the spread of taste, and the thickness of taste. It is a taste that is represented by changes in taste over time (afterglow or persistence). In the present invention, enhancing the richness includes, for example, imparting a richness to foods and drinks that do not have a richness, and enhancing the richness of foods and drinks that have a richness. The presence or absence and degree of richness can be evaluated by sensory evaluation by a specialized panel.

The beer-taste alcoholic beverage of the present invention preferably has a purine concentration of 0.01 to 10 mg / 100 mL.
For beer-taste alcoholic beverages having a purine concentration in the above range, the richness can be particularly enhanced by adding a sugar-modified protein.

The beer-taste alcoholic beverage of the present invention is a beer-taste beverage containing alcohol. The alcohol refers to ethanol, and the ethanol content is preferably 1% to 10% by volume, but is not particularly limited. Furthermore, the origin of the alcohol content contained in the beer-taste alcoholic beverage is not limited to fermented and non-fermented.

The manufacturing process of a general beer-taste alcoholic beverage is shown below.
First, in addition to wheat such as malt, if necessary, enzymes such as amylase are added to a mixture containing raw materials such as other grains, starch, sugars, bitterness agents, or coloring agents and water, and glue is added. It is saccharified and saccharified, and filtered to obtain a saccharified solution. If necessary, add hops and bitterness to the saccharified solution and boil, and remove solids such as coagulated protein in a clarification tank. As an alternative to this saccharified solution, hops may be added to a malt extract mixed with warm water and boiled. Hops may be mixed at any stage from the start of boiling to the end of boiling. As the conditions in the saccharification step, the boiling step, the solid content removing step and the like, known conditions may be used. As the conditions in the fermentation / liquor storage process and the like, known conditions may be used. The obtained fermentation broth is filtered, and carbon dioxide gas is added to the obtained filtrate. Then, it is filled in a container and sterilized to obtain a desired beer-taste alcoholic beverage. In each of the above steps, the richness enhancer may be added in any step up to filling.

A beer-taste alcoholic beverage produced without using malt as a raw material is a liquid obtained by mixing liquid sugar containing a carbon source, nitrogen source as an amino acid-containing material other than wheat or malt, hops, pigments, etc. with warm water. Use a sugar solution. The liquid sugar solution is boiled. When hops are used as a raw material, the hops may be mixed with the liquid sugar solution during boiling, not before the start of boiling. As an alternative to this saccharified solution, hops may be added to an extract using a raw material other than malt with warm water and boiled. Hops may be mixed at any stage from the start of boiling to the end of boiling. As the conditions in the fermentation / liquor storage process and the like, known conditions may be used. The obtained fermentation broth is filtered, and carbon dioxide gas is added to the obtained filtrate. Then, it is filled in a container and sterilized to obtain a desired beer-taste alcoholic beverage. In each of the above steps, the richness enhancer may be added in any step up to filling.

The non-fermented and alcohol-containing beer-taste alcoholic beverage may be one in which the alcohol content of the final product is adjusted by adding alcohol for raw materials or the like. The alcohol for raw materials may be added in any step from the saccharification step to the filling step.

The alcohol content of the beer-taste alcoholic beverage according to the present invention means the alcohol content (v / v%) in the beverage, and can be measured by any known method. It can be measured by a meter. Specifically, a sample from which carbon dioxide gas has been removed by filtration or ultrasonic waves from the beverage is prepared, and the sample is distilled by direct fire to measure the density of the obtained distillate at 15 ° C. Converted using "Table 2 Alcohol content and density (15 ° C) and specific gravity (15/15 ° C) conversion table" which is an attached table of (Heisei 19 National Tax Agency Instruction No. 6, revised on June 22, 2007). Can be obtained. When the alcohol content is low, less than 1.0%, a commercially available alcohol measuring device or gas chromatography may be used.

Fatty alcohol may be added to the beer-taste alcoholic beverage according to the present invention from the viewpoint of imparting a feeling of alcohol. The aliphatic alcohol is not particularly limited as long as it is known, but an aliphatic alcohol having 4 to 5 carbon atoms is preferable. In the present invention, preferred aliphatic alcohols include 2-methyl-1-propanol and 1-butanol as those having 4 carbon atoms, and 3-methyl-1-butanol and 1-pen as those having 5 carbon atoms. Examples thereof include tanol and 2-pentanol. These can be used in one type or a combination of two or more types.
The content of the aliphatic alcohol having 4 to 5 carbon atoms is preferably 0.0002 to 0.0007% by mass, and more preferably 0.0003 to 0.0006% by mass. In the present specification, the content of the aliphatic alcohol can be measured by using a headspace gas chromatograph method.

The sugar contained in the beer-taste alcoholic beverage according to the present invention means a sugar based on the nutrition labeling standard for foods (Ministry of Health, Labor and Welfare Notification No. 176, 2003). Specifically, sugar refers to food obtained by removing proteins, lipids, dietary fiber, ash, alcohol and water. The amount of sugar in a food is calculated by subtracting the amounts of protein, fat, dietary fiber, ash and water from the weight of the food. In this case, the amounts of protein, fat, dietary fiber, ash and water are measured by the methods listed in the nutrition labeling standards. Specifically, the amount of protein is measured by the nitrogen quantitative conversion method, the amount of lipid is measured by the ether extraction method, the chloroform / methanol mixed solution extraction method, the gelbell method, the acid decomposition method or the Reesegotleave method, and the amount of dietary fiber is measured. Is measured by high-speed liquid chromatograph method or Proski method, the amount of ash is measured by magnesium acetate addition ashing method, direct ashing method or sulfuric acid addition ashing method, and the amount of water is measured by Karl Fisher method, drying aid. Measure by the method, vacuum overheating drying method, normal pressure heating drying method or plastic film method.

The beer-taste alcoholic beverage according to the present invention may be low-carbohydrate in accordance with recent tastes for low-carbohydrate. Therefore, the sugar content of the beer-taste alcoholic beverage according to the present invention may be less than 2.5 g / 100 mL or less than 0.5 g / 100 mL. Although the lower limit is not particularly set, it is usually about 0.1 g / 100 mL, and may be, for example, 0.15 g / 100 mL or more or 0.2 g / 100 mL or more.

In the beer-taste alcoholic beverage according to the present invention, hops can be used as a part of the raw material.
When hops are used, ordinary pellet hops, powdered hops, and hop extracts used in the production of beer and the like can be appropriately selected and used according to a desired flavor. Further, processed hop products such as isometric hops and reduced hops may be used. These are included in the hops used in the beer-taste alcoholic beverage according to the present invention. The amount of hops added is not particularly limited, but is typically about 0.0001 to 1% by weight based on the total amount of the beverage.

The beer-taste alcoholic beverage according to the present invention may use other raw materials as needed, as long as the effects of the present invention are not impaired. For example, sweeteners (including high-sweetness sweeteners), bitterness, flavors, yeast extracts, colorants such as caramel pigments, plant-extracted saponin-based substances such as soybean saponin and kiraya saponin, plant proteins such as corn and soybean, and Peptide-containing substances, protein-based substances such as bovine serum albumin, seasonings such as dietary fiber and amino acids, and antioxidants such as ascorbic acid can be used as necessary as long as they do not interfere with the effects of the present invention.

The beer-taste alcoholic beverage according to the present invention can be packaged. The form of the container is not limited at all, and it can be filled in a sealed container such as a bottle, a can, a barrel, or a PET bottle to obtain a beverage in a container.

The rich taste enhancer of the present invention is a beer containing a sugar-modified protein having a molecular weight of 35 to 50 kDa in which the amino group of lysine constituting a grain-derived protein is glycated, and the proportion of malt in the raw material is less than 50% by weight. It is a rich taste enhancer for taste alcoholic beverages.

The richness enhancer of the present invention is a sugar-modified protein itself or a composition containing a sugar-modified protein, which is used to enhance the richness of a beer-taste alcoholic beverage.
The sugar-modified protein is a protein having a molecular weight of 35 to 50 kDa in which the amino group of lysine constituting the grain-derived protein is glycated, and the details thereof are as described above.
When the richness enhancer is a composition containing a sugar-modified protein, a known additive that can be added to a beverage may be optionally contained as long as the effect of the present invention is not impaired.

The content of glycated lysine in the sugar-modified protein contained in the rich taste enhancer of the present invention is 0.04 nmol / μg or more as the content (nmol) of glycated lysine per 1 μg of the sugar-modified protein. It is preferably 0.10 nmol / μg or more, and more preferably 0.10 nmol / μg or more.
Since such a sugar-modified protein contains a large amount of glycated lysine in the sugar-modified protein, it should be used as a richness enhancer capable of enhancing the richness of beer-taste alcoholic beverages with a small amount of use. Can be done.

The rich taste enhancer of the present invention can be obtained by purifying from beer brewed from North American malt, which contains a large amount of sugar-modified protein having a molecular weight of 35 to 50 kDa in which the amino group of lysine constituting a grain-derived protein is glycated. Can be done.
It can also be obtained by purifying from malt or barley containing a sugar-modified protein having a molecular weight of 35 to 50 kDa in which the amino group of lysine constituting a grain-derived protein is glycated.
Further, the protein may be extracted by mashing so as to contain a large amount of sugar-modified protein having a molecular weight of 35 to 50 kDa in which the amino group of lysine is glycated.

The method for producing a beer-taste alcoholic beverage of the present invention is not particularly limited, but includes a step of setting the ratio of malt in the raw material to less than 50% by weight and increasing the content of a sugar-modified protein having a molecular weight of 35 to 50 kDa. The manufacturing method is exemplified.
More specifically, a sugar-modified protein having a molecular weight of 35 to 50 kDa in which the amino group of lysine constituting a grain-derived protein is glycated is added to a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight. By doing so, there is a method of obtaining a beer-taste alcoholic beverage having an enhanced richness.
The method for producing a beer-taste alcoholic beverage by this method is also a method for enhancing the richness of the beer-taste alcoholic beverage of the present invention.

When a rich taste enhancer is added to a beer-taste alcoholic beverage, the amount of the rich taste enhancer added is such that the content of glycated lysine contained in the sugar-modified protein is 30.0 nmol / mL or more. Is preferable.
Further, the content of glycated lysine contained in the sugar-modified protein is preferably 50.0 nmol / mL or more, preferably 80.0 nmol / mL or more, and 100 nmol / mL or more. It is also preferable that the content is mL or more.
In this case, the amount of the richness enhancer added is preferably 7 μg / mL or more as the amount of the sugar-modified protein.

(Purification of sugar-modified protein derived from beer)
North American malt was crushed to an appropriate particle size, placed in a preparation tank, and then mixed with warm water. The starch was kept at a temperature suitable for malt enzyme for a sufficient time for saccharification, and starch was converted into sugar by the action of malt enzyme to produce a saccharified solution. Then, the saccharified solution was filtered, hops were added, and the mixture was boiled to produce hot wort to prepare for fermentation. The hot wort was cooled and brewer's yeast was added to it. In a few days, yeast worked to break down the sugar in the wort into alcohol and carbon dioxide to produce young beer. The young beer was transferred to a liquor tank and stored at low temperature for several tens of days. During this time, the beer was aged to harmonize the taste and aroma of the beer. After aging, the beer was filtered and bottled.
The sugar-modified protein was purified from beer brewed from North American malt by the above method according to the following.
(1) Fractionation with cation exchange resin 50 mL of cation exchange resin SP Sepharose was placed in an empty column. Beer brewed from North American malt was adsorbed on the resin. Then, the resin was transferred to a column and washed with 20 mM sodium acetate buffer (pH 4.5). Then, the fraction was collected by eluting with 20 mM sodium acetate (pH 4.5) + 0.5 M-NaCl. The obtained fraction was evaluated by SDS-PAGE, and the fraction containing a sugar-modified protein of 40 kDa was collected and used as a cation exchange resin-bound fraction.
(2) Extrafiltration (buffer exchange)
The cation exchange resin-bonded fraction obtained in (1) was centrifuged in a water-washed ultrafiltration unit (Amicon Ultra-15 30K manufactured by Merck) at 3500 rpm and ultrafiltered to obtain a concentrated solution.
(3) The concentrated solution obtained in the ammonium sulfate fraction (2) was added dropwise to 20 mM phosphate buffer (pH 7.0) + 2M ammonium sulfate, and the mixture was stirred. The suspension was then centrifuged (2330 g, 10 minutes). The supernatant was collected in a separate container. The collected solution was concentrated using an ultrafiltration unit. Add 20 mM sodium acetate (pH 4.5) to the concentrate, centrifuge (2330 g, 10 minutes), concentrate, and concentrate the sugar-modified protein assay (Bradford quantification (bovine serum albumin (BSA) equivalent), 20.4 mg / mL, 2.21 mL) was obtained. The purity of the obtained refined sugar-modified protein product was confirmed by SDS-PAGE.

(Purification of sugar-modified protein derived from barley)
The sugar-modified protein was purified from North American barley according to the following.
(1) Threshing of barley 451.88 g of North American barley was threshed with a rice mill (SATAKE Gabamil RSKM 3D) to obtain 394.02 g of barley.
(2) Freezing and crushing of refined barley 394.02 g of refined wheat was put into a ball mill, cooled with liquid nitrogen, and then crushed (1100 rpm, 10 minutes twice) to obtain a crushed barley product.
(3) Preparation of Barley Extract The crushed barley was added to an Erlenmeyer flask containing 5 L of water and stirred for 1 hour. Then, the crushed barley suspension was placed in a centrifugal tube and centrifuged at 4 ° C. and 8000 rpm for 10 minutes, and the centrifugal supernatant was used as a barley extract.
(4) Ammonium sulfate fraction (4-1) 40% saturated ammonium sulfate-treated barley extract is placed in a beaker, 243 g of ammonium sulfate is added per liter while stirring with a stirrer bar, and the mixture is stirred for 1 hour, and then the contents of the beaker are centrifuged. The mixture was centrifuged at 8000 rpm for 10 minutes at 4 ° C., and 4.9 L of the centrifugal supernatant was recovered to prepare a 40% saturated ammonium sulfate-treated fraction.
(4-2) 40% -60% Saturated Ammonium Sulfate Treatment 40% Saturated Ammonium Sulfate Treatment Put the entire fraction into a beaker, add 123 g of ammonium sulfate per liter while stirring with a stirrer bar, stir for 1 hour, and then stir the contents of the beaker. The mixture was placed in a centrifuge tube and centrifuged at 4 ° C. for 10 minutes at 8000 rpm, and the precipitated fraction was collected to prepare an ammonium sulfate fraction of the barley extract.
(4-3) Buffer exchange of ammonium sulfate fractions 100 mL of 20 mM sodium phosphate buffer (pH 6.5) was added to the ammonium sulfate fractions of the malt extract. Then, ultrafiltration was performed by an ultrafiltration unit (Amicon Ultra-15 30K manufactured by Merck), and buffer exchange was performed using a 20 mM phosphate buffer (pH 6.5).
(5) Anion Resin Fraction Anion Resin Q Sepharose 300 mL was packed in an econocolumn.
300 mL of Q Sepharose was added to the ammonium sulfate fraction of the barley extract, and batch adsorption was performed.
Then, the contents of the beaker were packed in a column and the pass-through fractions were collected (450 mL).
Then, 3000 mL of 20 mM phosphate buffer (pH 6.5) was loaded.
Then, 900 mL of 20 mM phosphate buffer (pH 6.5) containing 0.1 M NaCl, 900 mL of 20 mM phosphate buffer (pH 6.5) containing 0.2 M NaCl, and 20 mM phosphate buffer (pH 6.5) containing 0.3 M NaCl. ) 900 mL was loaded. Fractions were collected every 300 mL and analyzed by SDS-PAGE, and the fraction containing 40 kDa was collected and used as an anion exchange resin bound fraction.
Further, the concentrate was suspended in as little 20 mM sodium acetate buffer (pH 4.5) as possible, and 20 mM acetate buffer (pH 4.5) was added until the turbidity was removed.
Then, using an ultrafiltration unit (Amicon Ultra-15 30K manufactured by Merck), buffer exchange was carried out using a 20 mM sodium acetate buffer (pH 4.5) by ultrafiltration to obtain a sugar-modified protein derived from barley (a barley-derived sugar-modified protein). 1.14 mg / mL, 16.1 mL).

(Modification analysis of sugar-modified protein)
(1) Preparation of gel pieces 10 μL of purified sugar-modified protein solution (concentration 1 mg / mL, 20 mM sodium acetate buffer (pH 4.5)) was electrophoresed on SDS-PAGE, stained with CBB (Coomassie Brilliant Blue), and then in the vicinity of 40 kDa. A gel of sugar-modified protein was cut out.

(2) Enzymatic digestion of protein The cut out gel pieces were cut into small pieces, reduced with dithiothreitol (56 ° C. for 1 hour), and carbamid methylated with iodoacetamide (light-shielded, room temperature, 45 minutes). Then, 15 μL of 10 ng / μL chymotrypsin solution (5 mM calcium chloride, 50 mM ammonium hydrogen carbonate solution) containing 0.01% Protein Max, 15 μL of 5 mM calcium chloride, and 15 μL of 50 mM ammonium hydrogen carbonate solution were added and incubated overnight, and then the enzyme digestive juice was recovered. .. The recovered solution was dried under reduced pressure and redissolved in a 0.1% formic acid solution.

(3) Measurement by LC-MS / MS The measurement of LC-MS / MS was performed under the following conditions.
Equipment used: Direct flow nanoLC system Easy-nLC 1000TM (Thermo Scientific)
Trap column: Acclim PepMap® (Thermo Scientific)
Analytical column: NANO HPLC CAPILLARY COLUMN (Nikkyo Technos Co., Ltd.)
Liquid Chromatograph Mass Spectrometer Q Active Plus (Thermo Scientific)
Mobile phase: Liquid A: 0.1% formic acid / water, Liquid B: 0.1% formic acid / acetonitrile Flow rate: 300 nL / min
Gradient: 0-40% B / 0-30min, 40-60% B / 30-35min, 60-90% B / 35-37min, 90% B / 37-45min
Injection volume: 10 μL
Ionization mode: ESI Positive
Measurement range: MS1 (m / z 350-1750)
Data Dependent Scan mode

(4) Protein and modification analysis Protein identification and modification search were performed under the following conditions.
Search software: Proteome Discoverer 2.2.0.3888 (manufactured by Thermo Fisher)
Species: Barley (Hordeum vulgare), Hops (Humulus), Yeast (Saccharomyces cerevisiae)
Search conditions: Digestive enzymes: Chymotrypsin
Modification (Static): Carbamidethyl (Cysteine)
Modification (Dynamic): Oxidation (Methionine), Hex (K, R, Protein N-term), Hex (2) (K, R, Protein N-term), Hex (3) (K, R, Protein N-term) ), Acetyl (Protein N-term)
Precursor ion mass error range: Monoisotopic, ± 10 ppm
Product ion mass error range: ± 0.02 Da
Maximum number of miscribbage: 5
Confidence level (Percolator): High (the most probable level of the three levels of certainty)
Database: SwissProt

As a result of modification analysis, it was confirmed that the amino group of lysine of the sugar-modified protein was glycated.

(Evaluation of sugar modification degree of sugar-modified protein)
The degree of sugar modification of the purified sugar-modified protein was colorimetrically quantified by the NBT method using a Fluctosamine Assay kit (Abcam product, ab228558).
The degree of sugar modification of the barley-derived purified sugar-modified protein and the beer-derived purified sugar-modified protein was determined.
The number of tests for each protein was set to n = 2, and the average value was adopted.
The degree of sugar modification per weight of each purified sugar-modified protein is shown in Table 3.

(Measurement of concentration of glycated lysine in commercial beer-taste alcoholic beverages)
The concentration of glycated lysine in the commercially available beer-taste alcoholic beverages A and B having a malt ratio of less than 50% by weight was measured according to the following.
(1) Purification of sugar-modified proteins from commercial beer-taste alcoholic beverages A and B Similar to the purification of sugar-modified proteins from beer brewed from North American malt, sugar-modified proteins from commercial beer-taste alcoholic beverages A and B. Purification was performed.
A sugar-modified protein solution was obtained from a commercially available beer-taste alcoholic beverage A. A sugar-modified protein solution was obtained from a commercially available beer-taste alcoholic beverage B.
(2) Evaluation of sugar modification degree of sugar-modified protein purified from commercial beer-taste alcoholic beverages A and B The sugar modification degree of sugar-modified protein purified from commercial beer-taste alcoholic beverages A and B is barley from North America and malt from North America. It was measured by the NBT method in the same manner as the method for measuring the degree of sugar modification of the sugar-modified protein purified from beer brewed from.
(3) Measurement of sugar-modified protein concentration in commercial beer-taste alcoholic beverages The concentration of sugar-modified protein in beer of commercial beer-taste alcoholic beverages A and B is determined by using a LabChipTM GXII device (manufactured by Perkin Elmer) using an HT Protein Express chip. , Performed according to the standard protocol (n = 2).
(4) Calculation of the concentration of glycated lysine in the commercial beer-taste alcoholic beverages The concentration of glycated lysine in the commercial beer-taste alcoholic beverages A and B was calculated according to the following formula.
(Concentration of glycated lysine per protein weight of sugar-modified protein) × (Concentration of glycated lysine in beer-taste alcoholic beverage) = Concentration of glycated lysine in beer-taste alcoholic beverage Commercial beer-taste alcoholic beverage A The concentration of glycated lysate in was 28.8 nmol / mL, and the concentration of glycated lysine in the commercial beer-taste alcoholic beverage B was 11.6 nmol / mL.

(Evaluation of richness when sugar-modified protein is added to commercially available beer-taste alcoholic beverage A)
A beer-derived purified sugar-modified protein was added to a commercially available beer-taste alcoholic beverage A, and a sensory evaluation of richness was performed.
The purine concentration of the commercially available beer-taste alcoholic beverage A is 4.06 mg / 100 mL. The total purine concentration was determined by the decomposition method at the Japan Food Research Laboratories (the same applies hereinafter).
For the sensory evaluation, the rich taste of the commercially available beer-taste alcoholic beverage A to which no sugar-modified protein was added was set as a reference point of 0.9 points, and 25 μg / mL of beer-derived sugar-modified protein was added to the commercially available beer-taste alcoholic beverage A. The richness when added was set at 1.3 points as the reference point, and scored in 0.1 point increments by four expert panels.
Evaluation was performed by changing the addition concentration of the sugar-modified protein, and the results are shown in Table 1.
The richness was based on the following criteria.
0 points: Not felt at all 1 point: Slightly felt 2 points: Clearly felt 3 points: Very strongly felt Table 1 shows the content of glycated lysine contained in the sugar-modified protein in beer-taste alcoholic beverages. (Nmol / mL) is shown.

From the results shown in Table 1, a beer-taste alcoholic beverage was prepared by adding a sugar-modified protein to a commercially available beer-taste alcoholic beverage A having a malt ratio of less than 50% by weight to increase the content of glycated lysine. It can be seen that the richness of the beer can be enhanced.

(Evaluation of richness when sugar-modified protein is added to commercial beer-taste alcoholic beverage B)
A beer-derived sugar-modified protein was added to a commercially available beer-taste alcoholic beverage B, and a sensory evaluation of richness was performed.
The commercial beer-taste alcoholic beverage B is a beer-taste alcoholic beverage that is said to have less richness than the commercial beer-taste alcoholic beverage A.
The purine concentration of the commercially available beer-taste alcoholic beverage B is 4.2 mg / 100 mL.
The sensory evaluation was the same as the rich taste evaluation when a beer-derived sugar-modified protein was added to the commercially available beer-taste alcoholic beverage A.
When the rich taste of the commercially available beer-taste alcoholic beverage A to which the sugar-modified protein is not added is set to 0.9 as the reference point, and 25 μg / mL of the beer-derived sugar-modified protein is added to the commercially available beer-taste alcoholic beverage A. The richness is set at 1.3 points as the reference point.
Table 2 shows the content (nmol / mL) of glycated lysine contained in the sugar-modified protein in the beer-taste alcoholic beverage.

From the results shown in Table 2, even for beer-taste alcoholic beverages, which are originally considered to have a low richness, the richness is enhanced by adding a sugar-modified protein to increase the content of glycated lysine. I know I can do it.

(Effect of adding sugar-modified proteins with different degrees of sugar modification to beer)
A sugar-modified protein derived from barley and a sugar-modified protein derived from beer were added to the commercially available beer-taste alcoholic beverage A, respectively, and their effects on the rich taste were sensory-evaluated.
The degree of sugar modification of each purified protein was as shown in Table 3.

表４に示す通り、糖修飾タンパク質の添加量が同じであっても、糖修飾タンパク質の糖修飾度によってコク強度が異なり、糖修飾度に応じてコクが向上することが明らかになった。 The sensory evaluation was the same as the rich taste evaluation when a beer-derived sugar-modified protein was added to the commercially available beer-taste alcoholic beverage A.
Each sugar-modified protein was added to the commercially available beer-taste alcoholic beverage A and evaluated, and the results are shown in Table 4.
The amount of the sugar-modified protein added was 25 μg / mL.
Table 4 shows the content (nmol / mL) of glycated lysine contained in the sugar-modified protein in the beer-taste alcoholic beverage.

As shown in Table 4, it was clarified that even if the amount of the sugar-modified protein added was the same, the richness was different depending on the sugar-modified degree of the sugar-modified protein, and the richness was improved according to the sugar-modified degree.

According to the present invention, it is possible to provide a beer-taste alcoholic beverage having a strong rich taste by using a grain-derived raw material.

Claims (8)

A beer-taste alcoholic beverage in which the proportion of malt in the raw material is less than 50% by weight.
It contains a sugar-modified protein having a molecular weight of 35 to 50 kDa in which the amino group of lysine constituting a grain-derived protein is glycated.
A beer-taste alcoholic beverage having a glycated lysine content of 30.0 nmol / mL or more in the sugar-modified protein. The beer-taste alcoholic beverage according to claim 1, wherein the sugar-modified protein has a glycated lysine content of 35.3 nmol / mL or more. The beer-taste alcoholic beverage according to claim 1 or 2, wherein the sugar-modified protein has a glycated lysine content of 30.0 to 1000 nmol / mL. The beer-taste alcoholic beverage according to any one of claims 1 to 3, wherein the sugar glycated on the amino group of the glycated lysine is a monosaccharide or a disaccharide. The beer-taste alcoholic beverage according to any one of claims 1 to 4, wherein the grain is at least one selected from the group consisting of barley, wheat, corn, rice, and soybean. The beer-taste alcoholic beverage according to any one of claims 1 to 5, which does not contain wort. A rich taste enhancer for beer-taste alcoholic beverages, which comprises a sugar-modified protein having a molecular weight of 35 to 50 kDa and in which the amino group of lysine constituting a grain-derived protein is glycated, and the proportion of malt in the raw material is less than 50% by weight. Beer by adding a sugar-modified protein having a molecular weight of 35 to 50 kDa to which the amino group of lysine constituting a grain-derived protein is ligated to a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight. A method for enhancing the richness of beer-taste alcoholic beverages, which enhances the richness of taste alcoholic beverages.