Beer-flavored alcoholic beverage

Abstract

The purpose of the present invention is to provide a beverage having an enriched bulging taste, i.e., a beer-taste alcoholic beverage in which the proportion of malt in starting materials is 50 wt% or more and which has an enriched bulging taste, or a beer-taste alcoholic beverage in which the proportion of malt in starting materials is less than 50 wt% and which has an enriched bulging taste. The present invention pertains to: a beer-taste alcoholic beverage in which the proportion of malt in starting materials is 50 wt% or more and which has a 2'-O-methyladenosine concentration of 2 ppm or more; and a beer-taste alcoholic beverage in which the proportion of malt in starting materials is less than 50 wt% and which has a 2'-O-methyladenosine concentration of 1.5 ppm or more.

Description

Technical Field

[0001] The present invention relates to a beer-taste alcoholic beverage.

Background Art

[0002] With the diversification of consumers' preferences in recent years, the development of beer-taste alcoholic beverages having various flavor characteristics has been desired.

[0003] Patent Document 1 discloses that, in order to improve the flavor of a beer-taste alcoholic beverage, a peptide having a specific molecular weight is contained.

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0005] In Patent Document 1, as evaluation indices of a beverage, indices such as a beer-like taste, a smooth taste flow, and roughness remaining in the mouth are used, and it is described that a beverage having a high sensory evaluation score based on these indices can be obtained by containing a peptide of 10 to 20 kDa at a predetermined concentration.

[0006] Here, as a taste felt immediately after drinking a beer-taste beverage, there is a taste that cannot be expressed by the five basic tastes, that is, sweetness, saltiness, sourness, bitterness, and umami, and a taste having characteristics such as the intensity, spread, thickness, and balance of the taste strength of the taste being preferred. Such characteristics are referred to as "swelling" in this specification. It can be said that the beverage described in Patent Document 1 still has room for further improvement from the viewpoint of swelling, and a method for enhancing the swelling of a beer-taste alcoholic beverage has been desired.

[0007] The present invention aims to provide a beverage with enhanced fizziness. The present invention aims to provide a beer-taste alcoholic beverage with enhanced fizziness, wherein the ratio of malt in the raw materials is 50% by weight or more. In addition, the present invention aims to provide a beer-taste alcoholic beverage with enhanced fizziness, wherein the ratio of malt in the raw materials is less than 50% by weight. A beer-taste alcoholic beverage with a malt ratio of less than 50% by weight is a beverage for which enhancing fizziness is particularly desired because it tends to have poor fizziness.

Means for Solving the Problems

[0008] That is, the present invention relates to the following beer-taste alcoholic beverages. 〔1〕A beer-taste alcoholic beverage wherein the ratio of malt in the raw materials is 50% by weight or more, and the concentration of 2'-O-methyladenosine is 2 ppm or more. 〔2〕The beer-taste alcoholic beverage according to 〔1〕 above, wherein the concentration of 2'-O-methyladenosine is 2 to 10 ppm. 〔3〕Furthermore, the beer-taste alcoholic beverage according to 〔1〕 or 〔2〕 above, which further contains a protein having a molecular weight of 35 to 50 kDa, and the concentration of the protein is 10 ppm or more. 〔4〕The beer-taste alcoholic beverage according to 〔3〕 above, wherein the concentration of the protein is 200 ppm or less. 〔5〕A beer-taste alcoholic beverage wherein the ratio of malt in the raw materials is less than 50% by weight, and the concentration of 2'-O-methyladenosine is 1.5 ppm or more. 〔6〕The beer-taste alcoholic beverage according to 〔5〕 above, wherein the concentration of 2'-O-methyladenosine is 1.5 to 10 ppm. 〔7〕Furthermore, the beer-taste alcoholic beverage according to 〔5〕 or 〔6〕 above, which further contains a protein having a molecular weight of 35 to 50 kDa, and the concentration of the protein is 1 ppm or more. 〔8〕The beer - flavored alcoholic beverage according to the above - mentioned 〔7〕, wherein the concentration of the protein is 30 ppm or less.

Advantages of the Invention

[0009] According to the present invention, it is possible to provide a beer - flavored alcoholic beverage in which the ratio of malt in the raw materials is 50% by weight or more and the fullness is enhanced. Further, according to the present invention, it is possible to provide a beer - flavored alcoholic beverage in which the ratio of malt in the raw materials is less than 50% by weight and the fullness is enhanced.

Embodiments for Carrying Out the Invention

[0010] The beer - flavored alcoholic beverage which is the first embodiment of the present invention has a ratio of malt in the raw materials of 50% by weight or more. In the beer - flavored alcoholic beverage of the first embodiment of the present invention, the ratio of malt is preferably 100% by weight.

[0011] The beer - flavored alcoholic beverage which is the second embodiment of the present invention has a ratio of malt in the raw materials of less than 50% by weight. In the beer - flavored alcoholic beverage of the second embodiment of the present invention, the ratio of malt is preferably 30% by weight or more. Further, in the beer - flavored alcoholic beverage of the second embodiment of the present invention, the ratio of malt in the raw materials may be 0%.

[0012] Hereinafter, in this specification, the common description of the beer - flavored beverage in which the ratio of malt in the raw materials is 50% by weight or more, which is the first embodiment of the present invention, and the beer - flavored alcoholic beverage in which the ratio of malt in the raw materials is less than 50% by weight, which is the second embodiment of the present invention, will be simply described as the "beer - flavored alcoholic beverage".

[0013] Here, the "ratio of malt" refers to the ratio of the weight of malt in the raw materials other than water and hops, such as malt, rice, corn, sorghum, potato, starch, wheat other than malt, and sugars. However, components that can be added in trace amounts, such as acidulants, sweeteners, bittering agents, seasonings, and fragrances, are not included in the calculation of the above ratio.

[0014] The beer-taste alcoholic beverage of the present invention contains 2'-O-methyladenosine. 2'-O-methyladenosine is a kind of 2'-O-methylated ribonucleoside. In this specification, 2'-O-methyladenosine may be denoted as 2'OMA.

[0015] In the beer-taste alcoholic beverage in which the ratio of malt in the raw materials, which is the first form of the present invention, is 50% by weight or more, the concentration of 2'-O-methyladenosine is 2 ppm or more. When the concentration of 2'-O-methyladenosine is 2 ppm or more in a beer-taste alcoholic beverage in which the ratio of malt in the raw materials is 50% by weight or more, the swelling in the beer-taste alcoholic beverage in which the ratio of malt in the raw materials is 50% by weight or more can be enhanced. The fact that the content of 2'-O-methyladenosine at a predetermined concentration or more is related to the swelling in a beer-taste alcoholic beverage in which the ratio of malt in the raw materials is 50% by weight or more has not been known heretofore and is a finding discovered by the present inventors.

[0016] In the beer-taste alcoholic beverage in which the ratio of malt in the raw materials, which is the second form of the present invention, is less than 50% by weight, the concentration of 2'-O-methyladenosine is 1.5 ppm or more. When the concentration of 2'-O-methyladenosine is 1.5 ppm or more in a beer-taste alcoholic beverage in which the ratio of malt in the raw materials is less than 50% by weight, the swelling in the beer-taste alcoholic beverage in which the ratio of malt in the raw materials is less than 50% by weight can be enhanced. The fact that the content of 2'-O-methyladenosine at a predetermined concentration or more is related to the swelling in a beer-taste alcoholic beverage in which the ratio of malt in the raw materials is less than 50% by weight has not been known heretofore and is a finding discovered by the present inventors.

[0017] In the beer-taste alcoholic beverage of the first form of the present invention, it is preferable that the concentration of 2'-O-methyladenosine is 10 ppm or less. In a beer - flavored alcoholic beverage in which the ratio of malt in the raw material of the first form is 50% by weight or more, if the concentration of 2'-O - methyladenosine becomes too high, a strong pungent taste may be felt. In one aspect, the concentration of 2'-O - methyladenosine in the beer - flavored alcoholic beverage of the first form of the present invention is preferably 2 to 10 ppm.

[0018] In the beer - flavored alcoholic beverage of the second form of the present invention, the concentration of 2'-O - methyladenosine is preferably 10 ppm or less. In a beer - flavored alcoholic beverage in which the ratio of malt in the raw material of the second form is less than 50% by weight, if the concentration of 2'-O - methyladenosine becomes too high, an astringent taste may be felt. In the beer - flavored alcoholic beverage in which the ratio of malt in the raw material of the second form of the present invention is less than 50% by weight, the concentration of 2'-O - methyladenosine is preferably 5 ppm or more. In one aspect, the concentration of 2'-O - methyladenosine in the beer - flavored alcoholic beverage of the second form of the present invention is preferably 1.5 to 10 ppm, more preferably 5 to 10 ppm.

[0019] Further, in the beer - flavored alcoholic beverage in which the ratio of malt in the raw material of the first form of the present invention is 50% by weight or more, it further contains a protein having a molecular weight of 35 to 50 kDa, and the concentration of the above - mentioned protein is preferably 10 ppm or more.

[0020] Further, in the beer - flavored alcoholic beverage in which the ratio of malt in the raw material of the second form of the present invention is less than 50% by weight, it further contains a protein having a molecular weight of 35 to 50 kDa, and the concentration of the above - mentioned protein is preferably 1 ppm or more.

[0021] The protein with a molecular weight of 35 to 50 kDa is a protein found in the region of 35 to 50 kDa when a beer-taste alcoholic beverage is subjected to electrophoresis by SDS-PAGE. Before subjecting the beer-taste alcoholic beverage to electrophoresis by SDS-PAGE, for example, as a pretreatment, ultrafiltration may be performed on the beer-taste alcoholic beverage using a 30 kDa cut-off membrane. The above protein is preferably a protein with a molecular weight of 35 to 45 kDa, more preferably a protein of about 40 kDa. In this specification, the protein with a molecular weight of 35 to 50 kDa is also referred to as a 40 kDa protein.

[0022] The 40 kDa protein is preferably a cereal-derived protein. The above cereal is preferably at least one selected from the group consisting of barley, wheat, corn, rice, and soybeans. Also, when the cereal is wheat, it can contain a protein derived from known wheat used in the production of beer-taste alcoholic beverages. Examples of such wheat include barley, wheat, rye, crow wheat, oats, and emmer wheat, preferably barley. Also, it may be either germinated wheat or ungerminated wheat, preferably the malt of germinated wheat. These may be contained alone or in combination of two or more.

[0023] Also, as the 40 kDa protein, Serpin Z4 (alias: BSZ4, HorvuZ4, Major endosperm albumin or Protein Z) and Serpin Z7 (alias: BSZ7 or HorvuZ7) derived from barley (scientific name: Hordeum vulgare) are preferred. Also, in the above protein, a protein having an amino acid sequence in which some amino acids of the amino acid sequence are deleted, substituted, inserted, and / or added may be used.

[0024] By further containing a 40 kDa protein in addition to 2'-O-methyladenosine, the swelling in a beer-taste alcoholic beverage in which the ratio of malt in the raw material of the first form of the present invention is 50% by weight or more can be further enhanced. In the beer-taste alcoholic beverage of the first form of the present invention, the concentration of the 40 kDa protein is more preferably 25 ppm or more, further preferably 30 ppm or more, and preferably 200 ppm or less.

[0025] By further containing a 40 kDa protein in addition to 2'-O-methyladenosine, the swelling in a beer-taste alcoholic beverage in which the ratio of malt in the raw material of the second form of the present invention is less than 50% by weight can be further enhanced. In the beer-taste alcoholic beverage of the second form of the present invention, the concentration of the 40 kDa protein is more preferably 5 ppm or more, preferably 30 ppm or less, and more preferably 10 ppm or less.

[0026] Also, when both 2'-O-methyladenosine and a 40 kDa protein are contained, the swelling of the beer-taste alcoholic beverage can be effectively enhanced by the synergistic effect of 2'-O-methyladenosine and the 40 kDa protein.

[0027] The beer-taste alcoholic beverage of the present invention is a beer-taste beverage containing alcohol, and the alcohol concentration is preferably 1% (v / v) to 10% (v / v), but is not particularly limited. Further, the origin of the alcohol contained in the beer-taste alcoholic beverage is not limited to fermentation and non-fermentation. Here, the alcohol refers to ethanol and does not include the aliphatic alcohols described below. A beer-taste beverage refers to a carbonated beverage having a beer-like flavor.

[0028] The manufacturing process of a general beer-taste alcoholic beverage common to the first and second forms of the present invention is shown below. First, to a mixture containing malt such as barley, and other grains, starch, sugars, bittering agents, coloring agents, etc. as raw materials and water as needed, enzymes such as amylase are added as needed to perform gelatinization and saccharification, followed by filtration to obtain a saccharified liquid. As needed, hops, bittering agents, etc. are added to the saccharified liquid and boiled, and solid components such as coagulated proteins are removed in a clarification tank. As an alternative to this saccharified liquid, hot water may be added to malt extract, hops may be added and boiled. Hops may be mixed at any stage from the start of boiling to before the end of boiling. Conditions in the saccharification process, boiling process, solid component removal process, etc. may be those known conditions. Conditions in the fermentation and aging process, etc. may be those known conditions. The obtained fermented liquid is filtered, and carbon dioxide gas is added to the obtained filtrate. Then, it is filled into a container and passed through a sterilization process to obtain the target beer-taste alcoholic beverage.

[0029] The beer-taste alcoholic beverage in which the ratio of malt in the raw materials according to the second form of the present invention is less than 50% by weight may be produced by adding an alcohol beverage derived from wheat such as a distilled liquor derived from wheat (e.g., spirits, shochu, etc.) to the beer-taste alcoholic beverage produced by the method as described above.

[0030] When producing a beer-taste alcoholic beverage produced without using malt as an example of the beer-taste beverage according to the second form of the present invention, liquid sugar containing a carbon source, a nitrogen source as an amino acid-containing material other than wheat or malt, hops, pigments, etc. are mixed with warm water to form a liquid sugar solution. The liquid sugar solution is boiled. When using hops as a raw material, the hops may be mixed into the liquid sugar solution during boiling, rather than before the start of boiling. As an alternative to this saccharified liquid, hops may be added and boiled to a product obtained by adding warm water to an extract using raw materials other than malt. Hops may be mixed at any stage from the start of boiling to before the end of boiling. Conditions in the fermentation and aging process, etc. may be those known conditions. The obtained fermented liquid is filtered, and carbon dioxide gas is added to the obtained filtrate. Then, it is filled into a container and passed through a sterilization process to obtain the target beer-taste alcoholic beverage in which the ratio of malt in the raw materials is less than 50% by weight.

[0031] The non-fermented beer-taste alcoholic beverage containing alcohol may be one in which the alcohol content of the final product is adjusted by adding alcohol for raw materials or the like. The addition of alcohol for raw materials may be carried out in any step from the saccharification step to the filling step.

[0032] The alcohol degree of the beer-taste alcoholic beverage according to the present invention means the content (v / v%) of alcohol in the beverage and can be measured by any known method. For example, it can be measured by a vibrating densitometer. Specifically, a sample from which carbon dioxide gas has been removed by filtration or ultrasonic waves is prepared from the beverage, and the sample is distilled by direct fire. The density of the obtained distillate at 15 °C is measured, and it can be obtained by conversion using the "Table 2 Conversion Table of Alcohol Content, Density (15 °C), and Specific Gravity (15 / 15 °C)" which is an attached table of the specified analysis method of the National Tax Agency (National Tax Agency Ordinance No. 6 of 2007, revised on June 22, 2007). In the case of a low concentration with an alcohol degree of less than 1.0%, a commercially available alcohol measuring device or gas chromatography may also be used.

[0033] From the viewpoint of imparting a liquor feeling, an aliphatic alcohol may be added to the beer-taste alcoholic beverage according to the present invention. The aliphatic alcohol is not particularly limited as long as it is a known one, but an aliphatic alcohol having 4 to 5 carbon atoms is preferable. In the present invention, preferable aliphatic alcohols include, as those having 4 carbon atoms, 2-methyl-1-propanol, 1-butanol, etc., and as those having 5 carbon atoms, 3-methyl-1-butanol, 1-pentanol, 2-pentanol, etc. These can be used alone or in combination of two or more. The content of the aliphatic alcohol having 4 to 5 carbon atoms is preferably 0.0002 to 0.0007% by weight, more preferably 0.0003 to 0.0006% by weight. In this specification, the content of the aliphatic alcohol can be measured by the headspace gas chromatography method.

[0034] The carbohydrate contained in the beer-taste alcoholic beverage according to the present invention refers to the carbohydrate based on the Food Nutrition Labeling Standard (Ministry of Health, Labour and Welfare Notification No. 176 of 2003). Specifically, the carbohydrate refers to what remains after removing protein, lipid, dietary fiber, ash, alcohol content, and moisture from food. Also, the amount of carbohydrate in food is calculated by subtracting the amounts of protein, lipid, dietary fiber, ash, and moisture from the weight of the food. In this case, the amounts of protein, lipid, dietary fiber, ash, and moisture are measured by the methods listed in the nutrition labeling standard. Specifically, the amount of protein is measured by the nitrogen determination conversion method, the amount of lipid is measured by the ether extraction method, chloroform-methanol mixed solution extraction method, Gerber method, acid decomposition method, or Rose Gottlieb method, the amount of dietary fiber is measured by the high-performance liquid chromatography method or Prosky method, the amount of ash is measured by the magnesium acetate addition ashing method, direct ashing method, or sulfuric acid addition ashing method, and the amount of moisture is measured by the Karl Fischer method, drying aid method, reduced pressure superheated drying method, normal pressure heating drying method, or plastic film method.

[0035] The beer-taste alcoholic beverage according to the present invention may be low-carbohydrate in accordance with the recent low-carbohydrate preference. Therefore, the carbohydrate content of the beer-taste alcoholic beverage according to the present invention may be less than 2.5 g / 100 mL, or may be less than 0.5 g / 100 mL. Also, the lower limit is not particularly set, but is usually about 0.1 g / 100 mL. For example, it may be 0.15 g / 100 mL or more, or may be 0.2 g / 100 mL or more.

[0036] In the beer-taste alcoholic beverage according to the present invention, hop can be used as a part of the raw materials. When using hop, ordinary pellet hop, powder hop, and hop extract used in the production of beer or the like can be appropriately selected and used according to the desired flavor. Also, processed hop products such as isomerized hop and reduced hop may be used. The hop used in the beer-taste alcoholic beverage according to the present invention includes these. Also, the addition amount of hop is not particularly limited, but is typically about 0.0001 to 1% by weight based on the total amount of the beverage.

[0037] The beer-taste alcoholic beverage according to the present invention may, within the scope not interfering with the effects of the present invention, use other raw materials as necessary. For example, sweeteners (including high-intensity sweeteners), bittering agents, flavors, yeast extracts, coloring agents such as caramel pigments, plant-derived saponin substances such as soybean saponin and quillaja saponin, plant protein and peptide-containing substances such as corn and soybeans, protein 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 within the scope not interfering with the effects of the present invention.

[0038] The beer-taste alcoholic beverage according to the present invention can be packed in containers. The form of the container is not limited at all, and it can be filled into a sealed container such as a bottle, a can, a barrel, or a PET bottle to make a container-packed beverage.

[0039] The method for producing the beer-taste alcoholic beverage of the present invention is not particularly limited. For example, a method of adding a predetermined amount of 2'-O-methyladenosine to a beer-taste alcoholic beverage in which the ratio of malt in the raw materials of the first form of the present invention is 50% by weight or more or a beer-taste alcoholic beverage in which the ratio of malt in the raw materials of the second form of the present invention is less than 50% by weight is exemplified. Moreover, it is preferable to add a 40 kDa protein to the beer-taste alcoholic beverage of the first form or the second form of the present invention. The preparation of the 2'-O-methyladenosine and 40 kDa protein to be added can be carried out, for example, according to the procedures described in the examples below. Moreover, regarding 2'-O-methyladenosine and 40 kDa protein, their contents may be increased by adjusting various conditions in the production process of the beer-taste alcoholic beverage of the first form or the second form.

Examples

[0040] Hereinafter, the present invention will be specifically described by showing examples, but the present invention is not limited to the following examples.

[0041] (Purification of 2'-O-methyladenosine) Purification of 2'-O-methyladenosine (2'OMA) was carried out according to the following procedure. (1) Fractionation of beer by HP20 60 L of beer was fractionated using 10 L of Diaion® HP20 (manufactured by Mitsubishi Chemical Corporation). Before use, HP20 was washed three times with ethanol and then three times with 50% ethanol. The washed HP20 was packed into a column for large-scale fractionation and replaced with water. The same amount of distilled water was mixed with 60 L of degassed beer and flowed through the HP20 column using a medium-pressure pump. The solution that passed through the HP20 column was obtained as the flow-through fraction. 40 L of distilled water was flowed through using a medium-pressure pump, and the eluate was obtained as the water elution fraction. Similarly, 40 L each of aqueous ethanol (10% ethanol, 30% ethanol, and 70% ethanol) was flowed through, and the eluates were obtained as the 10% ethanol elution fraction, 30% ethanol elution fraction, and 70% ethanol elution fraction, respectively. Each elution fraction was refrigerated and stored as a dried product using an evaporator and a freeze dryer.

[0042] (2) Fractionation of the 30% ethanol elution fraction by LH-20 Among the HP20 fractions, the 30% ethanol elution fraction was fractionated using 1.2 kg of Sephadex® LH-20. LH-20 washed with ethanol was packed into a column for large-scale fractionation and replaced with water. Of the 30% ethanol elution fraction (87.9 g) obtained by HP20 fractionation, 17.6 g was dissolved in distilled water and applied to the LH-20 column. 13.5 L of distilled water was flowed through using a medium-pressure pump to obtain water elution fractions -1 to 6. Then, 7 L each of aqueous ethanol (35% ethanol, 70% ethanol, and 100% ethanol) was flowed through, and the eluates were obtained as the 35% ethanol elution fraction, 70% ethanol elution fraction, and 100% ethanol elution fraction, respectively. Each elution fraction was refrigerated and stored as a dried product using an evaporator and a freeze dryer.

[0043] (3) Separation of 2'OMA Of the 86.4 mg of the water elution fraction -4 (0.56 g) obtained by LH-20 fractionation, elution was carried out with 10% ethanol using HPLC (COSMOSIL 5C18-PAQ, 20×250 mm). Subsequently, the eluate from 13 min to 16 min was concentrated, and elution was carried out with a mixed solution of an ethanol-water (5:95 → 15:85) concentration gradient using HPLC (COSMOSIL 5C18-PAQ, 20×250 mm) to obtain compound (I) (5.0 mg, tR = 24 min). Compound (I) was identified as 2'-O-methyladenosine by analysis of the physical data of MS and NMR and comparison with the reference standard. The analytical instruments used were as follows. LC-MS; Q Exactive, manufactured by Thermo Fisher Scientific NMR; AVANCE400, manufactured by Bruker

[0044] (Purification of 40 kDa protein) Purification of the 40 kDa protein was carried out from commercially available beer (1 L) according to the following procedure.

[0045] (1) Fractionation by cation exchange resin 50 mL of the cation exchange resin SP Sepharose was placed in an empty column. The beer was adsorbed onto the resin. Subsequently, the resin was transferred to the column and washed with 20 mM sodium acetate buffer (pH 4.5). Then, elution was carried out with 20 mM sodium acetate (pH 4.5) + 0.5 M-NaCl, and the fractions were collected. The obtained fractions were evaluated by SDS-PAGE, and the fractions containing the 40 kDa protein were collected as the cation exchange resin-bound fraction.

[0046] (2) Ultrafiltration (buffer exchange) The cation exchange resin-bound fraction obtained in (1) was added 10 mL at a time to an ultrafiltration unit (Amicon Ultra-15 30K manufactured by Merck) washed with water, and centrifuged at 3500 rpm for ultrafiltration to obtain a concentrated solution.

[0047] (3) Ammonium sulfate fractionation 20 mM phosphate buffer (pH 7.0) + 2 M ammonium sulfate was placed in a beaker, and the concentrated solution obtained in (2) was added dropwise with stirring. Next, the suspension was centrifuged (2330 g, 10 minutes, room temperature). The supernatant was collected in a separate container. The collected solution was concentrated using an ultrafiltration unit. The concentrated solution was added to 20 mM sodium acetate (pH 4.5) and centrifuged (2330 g, 10 minutes, room temperature) for concentration to obtain a purified 40 kDa protein (Bradford quantification (equivalent to bovine serum albumin (BSA)), 20.4 mg / mL, 2.21 mL). The purity of the obtained 40 kDa protein was confirmed by SDS-PAGE.

[0048] After digesting the 40 kDa protein with an enzyme, an attempt was made to identify the protein by LC-MS / MS analysis. The band around 40 kDa separated by SDS-PAGE was cut out and subjected to reduction with dithiothreitol (56 °C, 1 hour) and carbamidomethylation with iodoacetamide (under light shielding, room temperature, 45 minutes). Then, 15 μL of a 10 ng / μL chymotrypsin solution containing 0.01% ProteaseMax (5 mM calcium chloride, 50 mM ammonium bicarbonate solution) and 15 μL of 5 mM calcium chloride, 50 mM ammonium bicarbonate solution were added and incubated overnight, after which the enzyme digestion solution was recovered. The recovered solution was dried under reduced pressure and redissolved in 0.1% formic acid solution. This was used for LC-MS / MS analysis.

[0049] (Measurement by LC-MS / MS) The LC-MS / MS measurement was performed under the following conditions. Equipment used: Direct Flow nanoLC system Easy-nLC 1000TM (Thermo Scientific) Trap column: Acclaim PepMap (registered trademark) (Thermo Scientific) Analysis column: NANO HPLC CAPILLARY COLUMN (Nikkyo Technos Co., Ltd.) Liquid chromatography mass spectrometer Q Exactive Plus (Thermo Scientific) Mobile Phase: Solvent A: 0.1% Formic Acid / Water, Solvent B: 0.1% Formic Acid / Acetonitrile Flow Rate: 300 nL / min Gradient: 0 - 40% B / 0 - 30 min, 40 - 60% B / 30 - 35 min, 60 - 90% B / 35 - 37 min, 90% B / 37 - 45 min Injection Volume: 10 μL Ionization Mode: ESI Positive Measurement Range: MS1 (m / z 350 - 1750) Data Dependent Scan Mode

[0050] (4) Analysis of Proteins Protein identification was performed under the following conditions. Search Software: Proteome Discoverer 2.2.0.388 (manufactured by ThermoFisher) Species: Barley (Hordeum vulgare), Hop (Humulus), Yeast (Saccharomyces cerevisiae) Search Conditions: Digestive Enzyme: Chymotrypsin Precursor Ion Mass Error Range: Monoisotopic, ±10 ppm Product Ion Mass Error Range: ±0.02 Da Maximum Number of Miss Cleavages: 5 Confidence Level (Percolator): High (the highest probability level among the three levels of certainty) Database: SwissProt

[0051] As a result, it was found that the 40 kDa protein is Barley - derived Serpin Z4 (sequence coverage rate: 77.2%) and Barley - derived Serpin Z7 (sequence coverage rate: 72.8%).

[0052] (Sensory Evaluation When 2’OMA is Added to Beer - flavored Alcoholic Beverages with a Ratio of Malt in Commercially Available Raw Materials of 50% by Weight or More) To the beer - flavored alcoholic beverage (A) with a malt ratio of 50% by weight or more in the commercially available raw materials, 2’OMA was added, and a sensory evaluation of the fullness was conducted. The beer - flavored alcoholic beverage (A) is a beer - flavored alcoholic beverage with a malt ratio of 50% by weight or more in the raw materials. The raw materials of the beer - flavored alcoholic beverage are malt and hops, and it contains 5.5% alcohol, 0.4 - 0.6 g of protein, 3.6 g of carbohydrates, and approximately 12.5 mg of purine per 100 ml as nutritional components.

[0053] The criteria for the sensory evaluation are as follows. Five professional panelists scored according to the following criteria in 0.05 - point increments, and the score values were averaged. The fullness intensity is based on the following criteria. 0 points: Not felt at all 1 point: Slightly felt 2 points: Clearly felt 3 points: Strongly felt As a reference point, the fullness of a commercially available beer - flavored alcoholic beverage (B) different from the above - mentioned commercially available beer - flavored alcoholic beverage (A) to be evaluated was set at 0.7 points as the reference beer - flavored alcoholic beverage (I). Also, the fullness of the same beer - flavored alcoholic beverage as the above - mentioned commercially available beer - flavored alcoholic beverage (A) to be evaluated was set at 1.5 points of the reference point as the reference beer - flavored alcoholic beverage (II). The reference beer - flavored alcoholic beverage (I) is a beer - flavored alcoholic beverage with a malt ratio in the raw materials exceeding 0% by weight and less than 50% by weight. The raw materials are sparkling wine, malt, hops, sugars, dietary fiber, and spirits (wheat), and it contains 4% alcohol, 0 - 0.2 g of protein, 0.5 - 0.8 g of carbohydrates, and approximately 2.0 mg of purine per 100 ml as nutritional components.

[0054] The procedure for the sensory evaluation is as follows. (1) Pour the beer - flavored alcoholic beverage (A) with a malt ratio of 50% by weight or more in the commercially available raw materials into a vial at 1 / 10 of the final volume (v / v). (2) Weigh and add 2’OMA in any weight. (3) Sonicate for 30 seconds. (4) Let stand at room temperature for 30 minutes. (5) Fill up the beer-taste alcoholic beverage to the final volume. (6) Dispense and evaluate by drinking.

[0055] (Analysis of commercially available beer-taste alcoholic beverage) The concentration of 2’OMA contained in the commercially available beer-taste alcoholic beverage used for sensory evaluation was quantified by LC-MS according to the following procedure. (1) Preparation of standard and calibration curve For 2’OMA, it was diluted to the following concentrations respectively, passed through a 0.22 μm filter, and then used for measurement. Final concentration: 0.001 ppm, 0.025 ppm, 0.050 ppm, 0.100 ppm, 0.200 ppm, 0.300 ppm, 0.500 ppm, 0.750 ppm, 1.000 ppm (1 ppm = 1 μg / mL) A 5% (v / v) ethanol aqueous solution was used as the diluent. In the analysis results of the standard, the measured value of the dilution ratio was adopted such that the measured value fell within the range where the linearity of the calibration curve was maintained (R 2 > 0.99).

[0056] The measurement conditions of LC-MS are as follows. LC-MS: X500R manufactured by AB SCIEX Separation column: HSST3 1.8 μm, 2.1 x 150 mm manufactured by Waters Eluent: Solution A: 0.1% formic acid / water, Solution B: 0.1% formic acid / acetonitrile Gradient: Solution A: Solution B = 98:2 → 2:98 (27 min) Injection volume: 5 μL Flow rate: 0.2 mL / min Column oven: 40 °C (MS) Ionization mode: ESI Positive Measurement range: MS1 (m / z 100 - 1000) Data Independent Scan mode Ion source temperature: 350 °C

[0057] (2) Preparation of measurement sample from beer - flavored alcoholic beverage with malt ratio of 50 wt% or more in commercially available raw materials The beer - flavored alcoholic beverage with malt ratio of 50 wt% or more in commercially available raw materials was degassed by sonication. After the bubbles subsided, it was diluted as appropriate, passed through a 0.22 - μm filter, and then used for measurement. As the diluent, a 5% (v / v) aqueous ethanol solution was used. The concentration of 2’OMA in the commercially available beer - flavored alcoholic beverage (A) was used as Control 1.

[0058] The concentration of the 40 - kDa protein in the beer - flavored alcoholic beverage with malt ratio of 50 wt% or more in commercially available raw materials used for sensory evaluation was measured according to the standard protocol using an HT Protein Express chip on a LabChipTM GXII device (manufactured by PerkinElmer) (n = 3). The concentration of the 40 - kDa protein in the beer - flavored alcoholic beverage (A) with malt ratio of 50 wt% or more in commercially available raw materials was 25 ppm.

[0059] (Example 1: Evaluation by addition of 2’OMA) The concentration of 2’OMA in the beer - flavored alcoholic beverage (A) (Control 1) with malt ratio of 50 wt% or more in commercially available raw materials was 1.549 ppm. In contrast, 2’OMA was added to make the 2’OMA concentration 2 ppm, 5 ppm, and 10 ppm respectively, and sensory evaluation was performed. The results of the sensory evaluation are shown in Table 1.

[0060]

Table 1

[0061] From the results shown in Table 1, it can be seen that for the beer-taste alcoholic beverage (A) in which the ratio of malt in the raw material is 50% by weight or more, the swelling is enhanced when the 2’OMA concentration is 2 ppm or more.

[0062] (Example 2: Evaluation of the synergistic effect of 2’OMA and 40 kDa protein) Based on a beverage obtained by adding 2’OMA to a commercially available beer-taste alcoholic beverage (A) in which the ratio of malt in the raw material is 50% by weight or more so that the concentration of 2’OMA is 2 ppm, the synergistic effect of 2’OMA and 40 kDa protein was evaluated by further adding 40 kDa protein. 40 kDa protein was added so that the concentration of 40 kDa protein was 30 ppm, 35 ppm, and 45 ppm, respectively. For the 40 kDa protein, the one purified above was used. Also, for comparison, an evaluation was made by adding only 5 ppm of 40 kDa protein to a commercially available beer-taste alcoholic beverage (A) in which the ratio of malt in the raw material is 50% by weight or more (comparative sample 1). The results of the sensory evaluation are shown in Table 2.

[0063]

Table 2

[0064] From the results shown in Table 2, it can be seen that by adding 2’OMA and further adding 40 kDa protein to a commercially available beer-taste alcoholic beverage (A) in which the ratio of malt in the raw material is 50% by weight or more, the swelling can be further enhanced.

[0065] (Example 3: Evaluation by adding 2’OMA) The concentration of 2’OMA contained in a commercially available beer-taste alcoholic beverage (C) (control 2) in which the ratio of malt in the raw material is 50% by weight or more was 1.01 ppm. The analysis of the commercially available beer-taste alcoholic beverage (C) was measured by the same method as in Example 1. On the other hand, 2’-OMA was added so that the 2’-OMA concentration became 2 ppm, and sensory evaluation was performed (Sample 7). Further, 2’-OMA was added to the above beer-taste alcoholic beverage (C) so that the 2’-OMA concentration became 2 ppm, and further, 40 kDa protein was added so that the 40 kDa protein concentration became 10 ppm, and sensory evaluation was performed (Sample 8). As the 40 kDa protein, the one purified above was used.

[0066] Note that the beer-taste alcoholic beverage (C) evaluated in Example 3 is a beer-taste alcoholic beverage in which the ratio of malt in the raw materials is 50% by weight or more. The beer-taste alcoholic beverage (C) is a beer-taste alcoholic beverage different from the commercially available beer-taste alcoholic beverage (A) used in Examples 1 and 2. The raw materials of the beer-taste alcoholic beverage (C) are malt, hop, rice, corn, and starch, and contain 5% alcohol, 0.2 to 0.4 g of protein, 3.0 g of carbohydrate, and 5 to 6 mg of purine per 100 ml as nutritional components.

[0067] The procedure for sensory evaluation is the same as the procedure in Example 1. The results of the sensory evaluation are shown in Table 3.

[0068]

Table 3

[0069] From the results shown in Table 3, it can be seen that even when the 40 kDa protein concentration is as low as 6.5 ppm, the addition of 2’-OMA to the beer-taste alcoholic beverage (C) in which the ratio of malt in the commercially available raw materials is 50% by weight or more can enhance the fullness. Further, it can be seen that when the 2’-OMA is 2 ppm and the 40 kDa protein concentration is 10 ppm, the sensory evaluation value is improved by 0.16, and the fullness of the beverage can be enhanced more effectively.

[0070] (Sensory evaluation when 2’-OMA is added to the beer-taste alcoholic beverage (B) in which the ratio of malt in the commercially available raw materials is less than 50% by weight) To the beer - flavored alcoholic beverage (B) with a malt ratio of less than 50% by weight in the commercially available raw materials, 2’OMA was added and a sensory evaluation of the fullness was conducted. The beer - flavored alcoholic beverage contains malt in the raw materials and is a beer - flavored alcoholic beverage with a malt ratio of less than 50% by weight in the raw materials. The raw materials are sparkling wine, malt, hops, saccharides, dietary fiber, spirits (wheat), and as nutritional components, it contains 4% alcohol, 0 - 0.2 g of protein, 0.5 - 0.8 g of carbohydrates, and approximately 2.0 mg of purine per 100 ml.

[0071] The criteria points for the sensory evaluation are as follows. Five professional panelists scored according to the following criteria in 0.05 - point increments, and the score values were averaged. The fullness intensity is based on the following criteria. 0 points: Not felt at all 1 point: Slightly felt 2 points: Clearly felt 3 points: Very strongly felt As a reference point, for the beer - flavored alcoholic beverage (I) which is the same as the beer - flavored alcoholic beverage (B) with a malt ratio of less than 50% by weight in the commercially available raw materials to be evaluated, the fullness was set at 0.7 points. Also, for another commercially available beer - flavored alcoholic beverage (A) with a malt ratio of 50% by weight or more in the raw materials, as the reference beer - flavored alcoholic beverage (II), the fullness was set at 1.5 times the reference point. The reference beer - flavored alcoholic beverage (II) is a beer - flavored alcoholic beverage with a malt ratio of 50% by weight or more in the raw materials. The raw materials are malt and hops, and as nutritional components, it contains 5.5% alcohol, 0.4 - 0.6 g of protein, 3.6 g of carbohydrates, and approximately 12.5 mg of purine per 100 ml.

[0072] The procedure for the sensory evaluation is as follows. Dispense a beer - flavored alcoholic beverage with a malt ratio of less than 50% by weight in the raw materials into a 1 / 10 - volume (v / v) vial of the final volume. Weigh and add 2’OMA in any weight. Perform sonication for 30 seconds. Let it stand at room temperature for 30 minutes. Fill up the beer - flavored alcoholic beverage to the final volume. Dispense and conduct a taste - evaluation.

[0073] (Analysis of a commercially available beer - flavored alcoholic beverage with a malt ratio of less than 50% by weight in the raw materials) The concentration of 2’OMA contained in the commercially available beer - flavored alcoholic beverage used for sensory evaluation was quantified by LC - MS according to the following procedure. (1) Preparation of standard samples and construction of a calibration curve For 2’OMA, dilute it to the following respective concentrations, pass it through a 0.22 - μm filter, and then use it for measurement. Final concentrations: 0.001 ppm, 0.025 ppm, 0.050 ppm, 0.100 ppm, 0.200 ppm, 0.300 ppm, 0.500 ppm, 0.750 ppm, 1.000 ppm (1 ppm = 1 μg / mL) As the diluent, use a 5% (v / v) aqueous ethanol solution. In addition, in the analysis results of the standard samples, adopt the measured value of the dilution ratio such that the measured value falls within the range (R 2 > 0.99) where the linearity of the calibration curve is maintained.

[0074] The measurement conditions of LC - MS are as follows. LC - MS: X500R manufactured by AB SCIEX Separation column: HSST3 1.8 μm, 2.1x150 mm manufactured by Waters Eluent: Solution A: 0.1% formic acid / water, Solution B: 0.1% formic acid / acetonitrile Gradient: Solution A:Solution B = 98:2 → 2:98 (27 min) Injection volume: 5 μL Flow rate: 0.2 mL / min Column oven: 40 °C (MS) Ionization mode: ESI Positive Measurement range: MS1 (m / z 100 - 1000) Data Independent Scan mode Ion source temperature: 350 °C

[0075] (2) Preparation of a measurement sample from a beer - flavored alcoholic beverage in which the ratio of malt in the commercially available raw material is less than 50% by weight The beer - flavored alcoholic beverage in which the ratio of malt in the commercially available raw material is less than 50% by weight was degassed by sonication. After the bubbles subsided, it was appropriately diluted, passed through a 0.22 - μm filter, and then used for measurement. As the diluent, a 5% (v / v) aqueous ethanol solution was used. The concentration of 2’OMA contained in the beer - flavored alcoholic beverage in which the ratio of malt in the commercially available raw material is less than 50% by weight was designated as Control 3.

[0076] (Example 4: Evaluation by addition of 2’OMA) The concentration of 2’OMA contained in the beer - flavored alcoholic beverage (Control 3) in which the ratio of malt in the commercially available raw material is less than 50% by weight was 0.347 ppm. In contrast, 2’OMA was added so that the 2’OMA concentration was 1.5 ppm, 5 ppm, and 10 ppm respectively, and a sensory evaluation was conducted. The results of the sensory evaluation are shown in Table 4.

[0077]

Table 4

[0078] From the results shown in Table 4, it can be seen that in the beer - flavored alcoholic beverage (B) in which the ratio of malt in the raw material is less than 50% by weight, when the 2’OMA concentration is 2 ppm or more, the swelling is enhanced.

[0079] (Example 5: Evaluation of the Synergistic Effect of 2’OMA and 40 kDa Protein) Based on a beer-taste alcoholic beverage (B) with a malt ratio of less than 50% by weight in commercially available raw materials, with 2’OMA added to make the concentration of 2’OMA 1.5 ppm, the synergistic effect of 2’OMA and 40 kDa protein was evaluated by further adding 40 kDa protein. The 40 kDa protein was added so that the concentration of the 40 kDa protein was 1 ppm, 5 ppm, and 10 ppm, respectively. For the 40 kDa protein, the one purified above was used. Also, for comparison, an evaluation was also conducted by adding only 5 ppm of 40 kDa protein to a beer-taste alcoholic beverage with a malt ratio of less than 50% by weight in commercially available raw materials (Comparative Sample 2). The results of the sensory evaluation are shown in Table 5.

[0080]

Table 5

[0081] From the results shown in Table 5, it can be seen that by adding 2’OMA and further adding 40 kDa protein to a beer-taste alcoholic beverage (B) with a malt ratio of less than 50% by weight in commercially available raw materials, the swelling can be further enhanced. From the results of Comparative Sample 2, it can be seen that adding only 40 kDa protein can enhance the swelling. However, for example, the increase value (0.20) from the control of the sensory evaluation in Sample 13 is larger than the additive effect (0.14) predicted when 2’OMA and 40 kDa protein are used in combination, which is obtained by adding the increase value (0.09) from the control of the sensory evaluation in the above Sample 9 and the increase value (0.05) from the control of the sensory evaluation in Comparative Sample 2. From this result, it can be said that when 2’OMA and 40 kDa protein are used in combination, a synergistic effect that cannot be expected is exerted in enhancing the swelling of a beer-taste alcoholic beverage (B) with a malt ratio of less than 50% by weight in commercially available raw materials.

Industrial Applicability

[0082] According to the present invention, in a beer-taste alcoholic beverage in which the ratio of malt in the raw materials is 50% by weight or more, a beverage with enhanced fullness can be provided. Further, according to the present invention, in a beer-taste alcoholic beverage in which the ratio of malt in the raw materials is less than 50% by weight, a beverage with enhanced fullness can be provided.

Claims

**Claim 1** A beer-taste alcoholic beverage in which the proportion of malt in the raw materials is 50% by weight or more, and the concentration of 2'-O-methyladenosine is 2 to 10 ppm. **Claim 2** Furthermore, it contains a protein with a molecular weight of 35 to 50 kDa, The beer-taste alcoholic beverage according to claim 1, wherein the concentration of the protein is 10 ppm or more. **Claim 3** The beer-taste alcoholic beverage according to claim 2, wherein the concentration of the protein is 200 ppm or less. **Claim 4** A beer-taste alcoholic beverage in which the proportion of malt in the raw materials is less than 50% by weight, and the concentration of 2'-O-methyladenosine is 1.5 to 10 ppm. **Claim 5** Furthermore, it contains a protein with a molecular weight of 35 to 50 kDa, The beer-taste alcoholic beverage according to claim 4, wherein the concentration of the protein is 1 ppm or more. **Claim 6** The beer-taste alcoholic beverage according to claim 5, wherein the concentration of the protein is 30 ppm or less.

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