Beer-taste alcoholic beverage

Abstract

To provide a beer-taste alcoholic beverage with enhanced body.SOLUTION: A beer-taste alcoholic beverage is such that the ratio of malt in the raw materials is 50 wt.% or more and the concentration of 2'-deoxyadenosine is 11 ppm or more. Furthermore, a beer-taste alcoholic beverage is provided containing a protein with a molecular weight of 35 to 50 kDa at a protein concentration of 10 ppm or more.SELECTED DRAWING: None

Description

[Technical Field]

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

[0002] With the recent diversification of consumer preferences, there is a demand for the development of beer-flavored alcoholic beverages with various flavor characteristics.

[0003] Patent Document 1 discloses that a peptide of a specific molecular weight is added to a beer-flavored alcoholic beverage in order to improve the flavor of the beverage. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Japanese Patent Application Laid-Open No. 2016-149975 Summary of the Invention [Problem to be solved by the invention]

[0005] Patent Document 1 uses indicators such as beer-like taste, smooth flow of flavor, and roughness remaining in the mouth as evaluation indices for beverages, and describes that by including a certain concentration of 10-20 kDa peptides, a beverage can be obtained that has a high sensory evaluation score based on these indices.

[0006] Here, the taste felt immediately after drinking a beer-taste beverage is often preferred as a taste that cannot be expressed by the five basic tastes, i.e., sweetness, saltiness, sourness, bitterness, and umami, and that has characteristics such as strong flavor, breadth, depth, long-lasting flavor, or a balanced flavor strength. These characteristics are referred to as "fullness" in this specification. The beverage described in Patent Document 1 can be said to have room for further improvement in terms of fullness, and a method for enhancing the fullness of beer-taste alcoholic beverages has been desired.

[0007] The present invention aims to provide a beverage with enhanced swelling. The present invention aims to provide a beer-taste alcoholic beverage with enhanced swelling, in which the malt content of the ingredients is 50% by weight or more. Another object of the present invention is to provide a beer-taste alcoholic beverage with enhanced swelling, in which the malt content of the ingredients is less than 50% by weight. Beer-taste alcoholic beverages with a malt content of less than 50% by weight tend to have poor swelling, so it is particularly desirable to enhance the swelling of these beverages. [Means for solving the problem]

[0008] That is, the present invention relates to the following beer-taste alcoholic beverages: [1] A beer-flavored alcoholic beverage in which the ratio of malt in the raw materials is 50% by weight or more and the concentration of 2'-deoxyadenosine is 11 ppm or more. [2] The beer-flavored alcoholic beverage according to [1] above, wherein the concentration of 2'-deoxyadenosine is 11 to 50 ppm. [3] The beer-flavored alcoholic beverage according to [1] or [2] above, further comprising a protein having a molecular weight of 35 to 50 kDa, the protein being present at a concentration of 10 ppm or more. [4] The beer-flavored alcoholic beverage according to [3] above, wherein the protein concentration is 200 ppm or less. [5] A beer-flavored alcoholic beverage in which the proportion of malt in the raw materials is less than 50% by weight, and in which the concentration of 2'-deoxyadenosine is 9.2 ppm or more. [6] The beer-flavored alcoholic beverage according to [5] above, wherein the concentration of 2'-deoxyadenosine is 9.2 to 100 ppm. [7] Further, the composition contains a protein having a molecular weight of 35 to 50 kDa, The beer-flavored alcoholic beverage according to [5] or [6] above, wherein the protein concentration is 1 ppm or more. [8] The beer-flavored alcoholic beverage according to [7] above, wherein the protein concentration is 30 ppm or less. [Effects of the Invention]

[0009] According to the present invention, a beer-taste alcoholic beverage having an enhanced fullness and containing 50% by weight or more of malt as an ingredient can be provided. Furthermore, according to the present invention, a beer-taste alcoholic beverage having an enhanced fullness can be provided in which the ratio of malt in the raw materials is less than 50% by weight. DETAILED DESCRIPTION OF THE INVENTION

[0010] The beer-taste alcoholic beverage according to the first embodiment of the present invention has a malt content of 50% by weight or more among its ingredients. In the beer-taste alcoholic beverage of the first embodiment of the present invention, the proportion of malt is preferably 100% by weight.

[0011] In the beer-taste alcoholic beverage according to the second embodiment of the present invention, the proportion of malt in the raw materials is less than 50% by weight. In the beer-taste alcoholic beverage of the second form of the present invention, the malt content is preferably 30% by weight or more. Alternatively, the beer-taste alcoholic beverage of the second form of the present invention may contain 0% malt in its ingredients.

[0012] Hereinafter, in this specification, explanations common to the beer-taste beverage of the first embodiment of the present invention, in which the proportion of malt in the ingredients is 50% by weight or more, and the beer-taste alcoholic beverage of the second embodiment of the present invention, in which the proportion of malt in the ingredients is less than 50% by weight, will be simply referred to as "beer-taste alcoholic beverage."

[0013] Here, "malt ratio" refers to the weight ratio of malt to the ingredients other than water and hops, including malt, rice, corn, sorghum, potatoes, starch, non-malted barley, and sugars. However, ingredients that may be added in trace amounts, such as acidulants, sweeteners, bittering agents, seasonings, and flavorings, are not included in the calculation of the above ratio.

[0014] The beer-taste alcoholic beverage of the present invention contains 2'-deoxyadenosine, a type of deoxyribonucleoside. Throughout this specification, 2'-deoxyadenosine may be abbreviated as 2'DA.

[0015] In a beer-taste alcoholic beverage according to the first embodiment of the present invention, in which the proportion of malt in the raw materials is 50% by weight or more, the concentration of 2'-deoxyadenosine is 11 ppm or more. In beer-flavored alcoholic beverages containing 50% or more by weight of malt as raw materials, a 2'-deoxyadenosine concentration of 11 ppm or higher can enhance the fullness of the beer-flavored alcoholic beverage. The correlation between the presence of a specific concentration or higher of 2'-deoxyadenosine and the fullness of a beer-flavored alcoholic beverage containing 50% or more by weight of malt as raw materials was previously unknown, and was discovered by the present inventors. In beer-flavored alcoholic beverages containing 50% or more by weight of malt as raw materials, the 2'-deoxyadenosine concentration is preferably 15 ppm or higher.

[0016] In the beer-taste alcoholic beverage of the first embodiment of the present invention, the concentration of 2'-deoxyadenosine is preferably 50 ppm or less. This is because if the concentration of 2'-deoxyadenosine is too high in a beer-flavored alcoholic beverage containing 50% or more by weight of malt as an ingredient, the aftertaste may be perceived as being too bitter and / or unpleasant. In one embodiment, the concentration of 2'-deoxyadenosine in the beer-taste alcoholic beverage of the first embodiment of the present invention is preferably 11 to 50 ppm, and more preferably 15 to 50 ppm.

[0017] In a beer-taste alcoholic beverage according to the second embodiment of the present invention, in which the proportion of malt in the raw materials is less than 50% by weight, the concentration of 2'-deoxyadenosine is 9.2 ppm or more. In beer-flavored alcoholic beverages whose raw materials contain less than 50% malt by weight, a 2'-deoxyadenosine concentration of 9.2 ppm or higher can enhance the fullness of the beer-flavored alcoholic beverage. The correlation between the inclusion of a certain concentration or higher of 2'-deoxyadenosine and the fullness of a beer-flavored alcoholic beverage whose raw materials contain less than 50% malt by weight was previously unknown and was discovered by the present inventors. In beer-flavored alcoholic beverages whose raw materials contain less than 50% malt by weight, the 2'-deoxyadenosine concentration is preferably 9.5 ppm or higher.

[0018] In a beer-taste alcoholic beverage according to the second embodiment of the present invention, in which the raw materials contain less than 50% malt by weight, the concentration of 2'-deoxyadenosine is preferably 100 ppm or less. This is because, in a beer-taste alcoholic beverage in which the raw materials contain less than 50% malt by weight, if the concentration of 2'-deoxyadenosine is too high, the aftertaste may be perceived as being strongly bitter. In a beer-taste alcoholic beverage in which the raw materials contain less than 50% malt by weight, the concentration of 2'-deoxyadenosine is more preferably 50 ppm or less. In one embodiment, the concentration of 2'-deoxyadenosine in a beer-taste alcoholic beverage in which the proportion of malt in the raw materials is less than 50 wt % is preferably 9.2 to 100 ppm, more preferably 9.5 to 100 ppm, and even more preferably 9.5 to 50 ppm.

[0019] The beer-taste beverage of the present invention preferably further contains a protein with a molecular weight of 35 to 50 kDa. A beer-taste alcoholic beverage according to the first embodiment of the present invention, in which the proportion of malt in the raw materials is 50% by weight or more, preferably further contains a protein with a molecular weight of 35 to 50 kDa, and the concentration of the protein is 10 ppm or more.

[0020] A beer-taste alcoholic beverage according to the second embodiment of the present invention, in which the proportion of malt in the raw materials is less than 50% by weight, preferably further contains a protein with a molecular weight of 35 to 50 kDa, and the concentration of the protein is 1 ppm or more.

[0021] Proteins with a molecular weight of 35 to 50 kDa are proteins found in the molecular weight range of 35 to 50 kDa when a beer-taste alcoholic beverage is subjected to SDS-PAGE electrophoresis. Prior to subjecting the beer-taste alcoholic beverage to SDS-PAGE electrophoresis, the beer-taste alcoholic beverage may be subjected to ultrafiltration using a 30 kDa cutoff membrane as a pretreatment, for example. The above protein preferably has a molecular weight of 35 to 45 kDa, more preferably about 40 kDa. In this specification, a 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 cereal is preferably at least one selected from the group consisting of barley, wheat, corn, rice, and soybeans. Furthermore, when the grain is barley, it may contain a protein derived from a known barley used in the production of beer-flavored alcoholic beverages. Examples of such barley include barley, wheat, rye, oats, oats, and oats, with barley being preferred. Furthermore, either germinated or ungerminated barley may be used, with germinated barley malt being preferred. These may be contained alone or in combination of two or more.

[0023] Preferred 40 kDa proteins include barley (scientific name: Hordeum vulgare)-derived Serpin Z4 (also known as BSZ4, HorvuZ4, Major endosperm albumin, or Protein Z) and barley-derived Serpin Z7 (also known as BSZ7 or HorvuZ7). The above proteins may also have an amino acid sequence in which some amino acids are deleted, substituted, inserted, and / or added.

[0024] By further including a 40 kDa protein in addition to 2'-deoxyadenosine, the fullness of a beer-flavored alcoholic beverage containing 50% by weight or more of malt as an ingredient can be further enhanced. Furthermore, in a beer-taste beverage according to the first embodiment of the present invention, in which the proportion of malt in the raw materials is 50% by weight or more, the concentration of the 40 kDa protein is preferably 25 ppm or more, even more preferably 30 ppm or more, and is preferably 200 ppm or less, even more preferably 100 ppm or less. In one aspect, in a beer-taste beverage according to the first embodiment of the present invention, the concentration of the 40 kDa protein is preferably 10 to 200 ppm, more preferably 25 to 200 ppm, even more preferably 30 to 200 ppm, and particularly preferably 30 to 100 ppm.

[0025] By including a 40 kDa protein in addition to 2'-deoxyadenosine, the fullness of the beer-taste alcoholic beverage of the second embodiment of the present invention, in which the proportion of malt in the raw materials is less than 50 wt %, can be further enhanced. In the beer-taste beverage of the second form of the present invention, the concentration of the 40 kDa protein is preferably 5 ppm or more, and preferably 30 ppm or less. In one embodiment, in the beer-taste beverage of the second form of the present invention, the concentration of the 40 kDa protein is preferably 1 to 30 ppm, and more preferably 5 to 30 ppm.

[0026] When both 2'-deoxyadenosine and the 40 kDa protein are contained, the fullness of the beer-flavored alcoholic beverage can be effectively enhanced due to the synergistic effect of 2'-deoxyadenosine 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. Furthermore, the origin of the alcohol contained in the beer-taste alcoholic beverage is not limited to fermented or non-fermented. Note that alcohol here refers to ethanol and does not include aliphatic alcohols, as described below. A beer-taste beverage is a carbonated beverage with a beer-like flavor.

[0028] Below, a general process for producing a beer-taste alcoholic beverage common to the first and second embodiments of the present invention is shown. First, a mixture containing malt and other barley, as well as other grains, starch, sugars, bittering agents, or coloring agents, and water is gelatinized and saccharified, optionally with the addition of enzymes such as amylase. The mixture is then filtered to produce a saccharified liquor. Hops and bittering agents are added as needed, and the saccharified liquor is boiled, with solids such as coagulated proteins removed in a clarifying tank. As an alternative to this saccharified liquor, hops can be added to malt extract and warm water, and then boiled. Hops can be added at any stage, from the start of boiling to the end of boiling. Known conditions can be used for the saccharification, boiling, and solids removal processes. Known conditions can be used for the fermentation and storage processes. The resulting fermented liquor is filtered, and carbon dioxide gas is added to the filtrate. The liquor is then filled into containers and sterilized to produce the desired beer-flavored alcoholic beverage.

[0029] A beer-taste alcoholic beverage having a malt ratio of less than 50% by weight among the ingredients, which is a second embodiment of the present invention, may be produced by adding a barley-derived alcoholic beverage such as a barley-derived distilled alcoholic beverage (e.g., spirits or shochu) to a beer-taste alcoholic beverage produced by the method described above.

[0030] When producing a beer-flavored alcoholic beverage produced without using malt as a raw material, which is an example of the beer-flavored beverage of the second embodiment of the present invention, a liquid sugar containing a carbon source, a nitrogen source as an amino acid-containing material other than barley or malt, hops, a color, etc. are mixed with warm water to form a liquid sugar solution. This liquid sugar solution is boiled. When hops are used as a raw material, hops may be added to the liquid sugar solution during boiling rather than before the start of boiling. As an alternative to this saccharified solution, hops may be added to an extract made from a raw material other than malt, to which warm water is added, and the mixture is then boiled. Hops may be added at any stage from the start of boiling to the end of boiling. Known conditions may be used for the fermentation and storage processes. The resulting fermented liquid is filtered, and carbon dioxide gas is added to the resulting filtrate. The beverage is then filled into containers and sterilized to obtain a beer-flavored alcoholic beverage having a malt content of less than 50% by weight.

[0031] A non-fermented, alcohol-containing beer-taste alcoholic beverage may be one in which the alcohol content of the final product is adjusted by adding a raw material alcohol, etc. The raw material alcohol may be added at any step from the saccharification step to the filling step.

[0032] The alcohol content (v / v%) of the beer-taste alcoholic beverage of the present invention refers to the alcohol content (v / v%) in the beverage and can be measured by any known method, for example, a vibration density meter. Specifically, the beverage is decarbonated by filtration or ultrasonication to prepare a sample, which is then subjected to direct flame distillation. The density of the resulting distillate is measured at 15°C and converted using "Table 2: Conversion Table of Alcohol Content, Density (15°C), and Specific Gravity (15 / 15°C)," an appendix to the National Tax Agency's Prescribed Analysis Methods (National Tax Agency Ordinance No. 6 of 2007, revised June 22, 2007). For low concentrations of less than 1.0% alcohol, a commercially available alcohol measuring device or gas chromatography may be used.

[0033] An aliphatic alcohol may be added to the beer-taste alcoholic beverage of the present invention in order to impart a boozy flavor. There are no particular limitations on the aliphatic alcohol as long as it is a known alcohol, but aliphatic alcohols with 4 to 5 carbon atoms are preferred. In the present invention, preferred aliphatic alcohols with 4 carbon atoms include 2-methyl-1-propanol and 1-butanol, and preferred aliphatic alcohols with 5 carbon atoms include 3-methyl-1-butanol, 1-pentanol, and 2-pentanol. These may 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, and more preferably 0.0003 to 0.0006% by weight. In this specification, the content of the aliphatic alcohol can be measured using headspace gas chromatography.

[0034] The carbohydrates contained in the beer-taste alcoholic beverage according to the present invention refer to the carbohydrates based on the Nutrition Labeling Standards for Foods (Ministry of Health, Labour and Welfare Notification No. 176 of 2003). Specifically, carbohydrates refer to the amount of food excluding protein, lipids, dietary fiber, ash, alcohol, and water. The amount of carbohydrates in a food is calculated by subtracting the amounts of protein, lipids, dietary fiber, ash, and water from the weight of the food. In this case, the amounts of protein, lipids, dietary fiber, ash, and water are measured using the methods set forth in the Nutrition Labeling Standards. Specifically, the amount of protein is measured using the nitrogen quantitative conversion method, the amount of lipids is measured using the ether extraction method, chloroform-methanol mixed liquid extraction method, Gerber method, acid hydrolysis method or Roese-Gottlieb method, the amount of dietary fiber is measured using high-performance liquid chromatography or the Prosky method, the amount of ash is measured using the magnesium acetate ashing method, direct ashing method or sulfuric acid ashing method, and the amount of moisture is measured using the Karl Fischer method, drying aid method, reduced-pressure superheat drying method, normal pressure heat drying method or plastic film method.

[0035] The beer-taste alcoholic beverage of the present invention may be low in carbohydrates in line with the recent trend toward low carbohydrate foods. Therefore, the carbohydrate content of the beer-taste alcoholic beverage of the present invention may be less than 2.5 g / 100 mL or less than 0.5 g / 100 mL. While there is no specific lower limit, it is typically around 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.

[0036] In the beer-taste alcoholic beverage according to the present invention, hops can be used as part of the ingredients. When using hops, typical pelleted hops, powdered hops, or hop extracts used in the production of beer and the like can be appropriately selected and used depending on the desired flavor. Also, processed hop products such as isomerized hops and reduced hops may be used. These products are included in the hops used in the beer-flavored alcoholic beverage of the present invention. The amount of hops added is not particularly limited, but is typically about 0.0001 to 1% by weight of the total amount of the beverage.

[0037] The beer-taste alcoholic beverage of the present invention may contain other ingredients as needed, provided that the effects of the present invention are not impaired. For example, sweeteners (including high-intensity sweeteners), bittering agents, flavorings, yeast extracts, coloring agents such as caramel color, plant-extracted saponin substances such as soybean saponin and quillaja saponin, plant protein and peptide-containing substances from corn, soybeans, and the like, protein-based substances such as bovine serum albumin, seasonings such as dietary fiber and amino acids, and antioxidants such as ascorbic acid may be used as needed, provided that the effects of the present invention are not impaired.

[0038] The beer-taste alcoholic beverage according to the present invention can be packaged in a container. The type of container is not particularly limited, and the beer-taste alcoholic beverage can be packaged in a sealed container such as a bottle, can, barrel, or PET bottle to form a packaged beverage.

[0039] The method for producing the beer-taste alcoholic beverage of the present invention is not particularly limited, but may include, for example, adding a predetermined amount of 2'-deoxyadenosine to a beer-taste alcoholic beverage whose raw materials contain 50% or more by weight of malt, or to a beer-taste alcoholic beverage whose raw materials contain less than 50% by weight of malt. Furthermore, it is preferable to add the 40 kDa protein to a beer-taste alcoholic beverage in which the ratio of malt in the raw materials is 50% by weight or more according to the first embodiment of the present invention, or to a beer-taste alcoholic beverage in which the ratio of malt in the raw materials is less than 50% by weight according to the second embodiment of the present invention.

[0040] The 2'-deoxyadenosine and 40 kDa protein to be added can be prepared, for example, by the procedures described in the Examples below. Furthermore, the contents of 2'-deoxyadenosine and 40 kDa protein may be increased by adjusting the conditions in the production process of the beer-taste alcoholic beverage. [Example]

[0041] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to the following examples.

[0042] (Purification of 2'-deoxyadenosine) 2'-Deoxyadenosine (2'DA) was purified as follows. (1) Fractionation of beer using HP20 60 L of beer was fractionated using 10 L of Diaion® HP20 (Mitsubishi Chemical Corporation). The HP20 was washed three times with ethanol and then three times with 50% ethanol before use. The washed HP20 was packed into a large-scale fractionation column and purged with water. 60 L of degassed beer was mixed with an equal volume of distilled water and pumped through the HP20 column using a medium-pressure pump. The solution that passed through the HP20 column was collected as the flow-through fraction. 40 L of distilled water was pumped through the column using a medium-pressure pump, and the eluate was collected as the water-eluted fraction. Similarly, 40 L of aqueous ethanol (10% ethanol, 30% ethanol, and 70% ethanol) was pumped through the column, and the eluates were collected as the 10% ethanol-eluted fraction, the 30% ethanol-eluted fraction, and the 70% ethanol-eluted fraction, respectively. Each elution fraction was dried using an evaporator and a freeze-dryer and stored refrigerated.

[0043] (2) LH-20 fraction from 30% ethanol elution fraction The 30% ethanol-eluted fraction from the HP20 fraction was fractionated using 1.2 kg of Sephadex® LH-20. The LH-20 was washed with ethanol and loaded onto a large-scale fractionation column, followed by water replacement. Of the 30% ethanol-eluted fraction (87.9 g) obtained from the HP20 fractionation, 17.6 g was dissolved in distilled water and applied to the LH-20 column. Using a medium-pressure pump, 13.5 L of distilled water was pumped through the column, yielding water-eluted fractions 1 to 6. Next, 7 L of aqueous ethanol (35% ethanol, 70% ethanol, and 100% ethanol) was pumped through the column, yielding the 35% ethanol-eluted fraction, 70% ethanol-eluted fraction, and 100% ethanol-eluted fraction, respectively. The eluted fractions were dried using an evaporator and a freeze-dryer and stored refrigerated.

[0044] (3) Separation of 2'DA A portion of the water-eluted fraction-4 (0.56 g) obtained from the LH-20 fractionation was eluted with 10% ethanol using HPLC (COSMOSIL 5C18-PAQ, 20 × 250 mm). The eluate from 10 min to 13 min was concentrated and eluted with a gradient mixture of ethanol and water (5:95 → 15:85) using HPLC (COSMOSIL 5C18-PAQ, 20 × 250 mm) to obtain Compound (I) (0.5 mg, tR = 21 min). Compound (I) was identified as 2'-deoxyadenosine by analysis of physical data from MS and NMR and comparison with authentic samples. The analytical instruments used are as follows: LC-MS;Q Exactive, manufactured by Thermo Fisher Scientific NMR: AVANCE400, manufactured by Bruker

[0045] (Purification of 40kDa protein) A 40 kDa protein was purified from commercially available beer (1 L) as follows.

[0046] (1) Fractionation by cation exchange resin 50 mL of cation exchange resin SP Sepharose was placed in an empty column. The beer was allowed to adsorb onto the resin. The resin was then transferred to a column and washed with 20 mM sodium acetate buffer (pH 4.5). Elution was then performed with 20 mM sodium acetate (pH 4.5) + 0.5 M NaCl, and the fractions were collected. The resulting fractions were evaluated by SDS-PAGE, and the fractions containing the 40 kDa protein were collected and used as the cation exchange resin-bound fraction.

[0047] (2) Ultrafiltration (buffer exchange) 10 mL of the cation exchange resin-bound fraction obtained in (1) was added to a water-washed ultrafiltration unit (Merck Amicon Ultra-15 30K), centrifuged at 3500 rpm, and ultrafiltered to obtain a concentrate.

[0048] (3) Ammonium sulfate fraction 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 and stirred. The suspension was then 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) to obtain a 40 kDa purified protein (Bradford quantitation (bovine serum albumin (BSA) equivalent), 20.4 mg / mL, 2.21 mL). The purity of the obtained 40 kDa protein was confirmed by SDS-PAGE.

[0049] The 40 kDa protein was digested with an enzyme and then analyzed by LC-MS / MS to attempt to identify the protein. A band around 40 kDa separated by SDS-PAGE was excised and subjected to reduction with dithiothreitol (56°C, 1 hour) and carbamidomethylation with iodoacetamide (protected from light, room temperature, 45 minutes). Next, 15 μL of 10 ng / μL chymotrypsin solution (5 mM calcium chloride, 50 mM ammonium bicarbonate solution) containing 0.01% Protease Max and 15 μL of 5 mM calcium chloride, 50 mM ammonium bicarbonate solution were added, and the mixture was incubated overnight. The enzyme digestion solution was then recovered. The recovered solution was evaporated to dryness under reduced pressure and redissolved in 0.1% formic acid solution. This was used for LC-MS / MS analysis.

[0050] (Measurement by LC-MS / MS) The LC-MS / MS measurements were carried out under the following conditions. Equipment used: Direct flow nanoLC system Easy-nLC 1000TM (Thermo Scientific) Trap column: Acclaim PepMap® (Thermo Scientific) Analytical column: NANO HPLC CAPILLARY COLUMN (Nikkyo Technos Co., Ltd.) Liquid chromatograph mass spectrometer Q Exactive Plus (Thermo Scientific) Mobile phase: Solution A: 0.1% formic acid / water, Solution 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

[0051] (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 miscleavages: 5 Confidence level (Percolator): High (the highest probability level among the three levels of certainty) Database: SwissProt

[0052] As a result, it was found that the 40 kDa protein was Serpin Z4 derived from barley (sequence coverage rate: 77.2%) and Serpin Z7 derived from barley (sequence coverage rate: 72.8%).

[0053] <I. Sensory evaluation when 2’DA is added to a commercially available beer - flavored alcoholic beverage in which the ratio of malt in the raw material is 50% by weight or more> In Examples 1 and 2, 2'-deoxyadenosine (2'DA) was added to a commercially available beer-taste alcoholic beverage (A), and a sensory evaluation of fullness was performed. The beer-flavored alcoholic beverage (A) is a beer-flavored alcoholic beverage containing 50% by weight or more of malt as its raw materials. The raw materials for this beer-flavored alcoholic beverage are malt and hops, and it contains 5.5% alcohol, 0.4 to 0.6 g of protein, 3.6 g of carbohydrates, and approximately 12.5 mg of purines per 100 ml.

[0054] The criteria for the sensory evaluation are as follows: Five expert panel members scored the items in increments of 0.05 points according to the following criteria, and the scores were averaged. The swelling strength is based on the following criteria: 0 points: Not felt at all 1 point: Slightly felt 2 points: Clear feeling 3 points: Very strong A commercially available beer-taste alcoholic beverage (B) different from the commercially available beer-taste alcoholic beverage (A) to be evaluated was used as the reference beer-taste alcoholic beverage (I), with a fullness score of 0.7 points. A commercially available beer-taste alcoholic beverage identical to the commercially available beer-taste alcoholic beverage (A) to be evaluated was used as the reference beer-taste alcoholic beverage (II), with a fullness score of 1.5 points. A beer-flavored alcoholic beverage (I) according to this standard is a beer-flavored alcoholic beverage in which the proportion of malt in the raw materials is greater than 0% by weight and less than 50% by weight. The ingredients are happoshu, malt, hops, sugar, dietary fiber, and spirits (wheat), and the nutritional content per 100ml is 4% alcohol, 0-0.2g protein, 0.5-0.8g carbohydrates, and approximately 2.0mg purines.

[0055] The procedure for the sensory evaluation was as follows. (1) Dispense the beer-flavored alcoholic beverage to be evaluated into vials at 1 / 10 of the final volume (v / v). (2) Weigh out an arbitrary amount of 2'DA and add it. (3) Sonicate for 30 seconds (4) Leave it at room temperature for 30 minutes. (5) Fill the container with the beer-flavored alcoholic beverage to the final volume. (6) Dispense and evaluate swallowing

[0056] (Analysis of commercially available beer-flavored alcoholic beverages) The concentrations of 2'DA contained in the commercially available beer-flavored alcoholic beverages used in the sensory evaluation were quantified by LC-MS using the following procedure. (1) Preparation of standard samples and creation of calibration curves 2'DA was diluted to the following concentrations, passed through a 0.22 μm filter, and then subjected to measurement. Final concentration: 0.001ppm, 0.025ppm, 0.050ppm, 0.100ppm, 0.200ppm, 0.300ppm, 0.500ppm, 0.750ppm, 1.000ppm (1 ppm = 1 μg / mL) The diluent used was a 5% (v / v) aqueous ethanol solution. In the analytical results of the standard, the range in which the linearity of the calibration curve is maintained (R 2 The measured values ​​were taken at a dilution ratio such that the measured values ​​fell within the range of 0.99 (>0.99).

[0057] The LC-MS measurement conditions are as follows. LC-MS: AB Sciex X500R Separation column: Waters HSST3 1.8 μm, 2.1 x 150 mm 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: 5μL Flow rate: 0.2mL / min Column oven: 40℃ (MS) Ionization mode: ESI Positive Measurement range: MS1 (m / z 100-1000) Data Independent Scan Mode Ion source temperature: 350℃

[0058] (2) Preparation of measurement samples from commercially available beer-flavored alcoholic beverages A commercially available beer-flavored alcoholic beverage was degassed by sonication, and after the bubbles had settled, it was diluted appropriately and passed through a 0.22 μm filter before being used for measurement. The diluent used was a 5% (v / v) aqueous ethanol solution. The concentration of 2'DA contained in a commercially available beer-flavored alcoholic beverage (A) was designated as Control 1.

[0059] The concentrations of 40 kDa proteins in the commercially available beer-flavored alcoholic beverages used in the sensory evaluation were measured using HT Protein Express chips on a LabChip™ GXII instrument (PerkinElmer) according to the standard protocol (n = 3). The concentration of the 40 kDa protein in a commercially available beer-flavored alcoholic beverage (A) was 25 ppm.

[0060] (Example 1: Evaluation by adding 2'DA) The concentration of 2'DA contained in a commercially available beer-flavored alcoholic beverage (A) (Control 1) was 8 ppm. To this, 2'DA was added so that the 2'DA concentrations were 11 ppm, 15 ppm, 20 ppm, and 50 ppm, and a sensory evaluation was performed. The results of the sensory evaluation are shown in Table 1.

[0061] [Table 1]

[0062] The results shown in Table 1 indicate that in beer-flavored alcoholic beverages in which the proportion of malt in the raw materials is 50% by weight or more, fullness is enhanced when the 2'DA concentration is 11 ppm or more.

[0063] Example 2: Evaluation of the synergistic effect of 2'DA and 40 kDa protein The synergistic effect of 2'DA and 40kDa protein was evaluated by adding 40kDa protein to a commercially available beer-flavored alcoholic beverage (A) containing 50% or more malt as the raw material, with 2'DA added to achieve a 2'DA concentration of 11 ppm. The 40kDa protein was added to achieve concentrations of 30 ppm, 35 ppm, and 45 ppm, respectively. The 40kDa protein used was purified as described above. For comparison, a commercially available beer-flavored alcoholic beverage (A) was also evaluated by adding 5 ppm of the 40 kDa protein alone (control sample 2-1). The results of the sensory evaluation are shown in Table 2.

[0064] [Table 2]

[0065] The results shown in Table 2 indicate that the volume of a commercially available beer-flavored alcoholic beverage (A), whose raw material contains malt at a ratio of 50% by weight or more, can be further enhanced by adding 2'DA and further adding 40 kDa protein. The results for comparison sample 2-1 indicate that the addition of 40 kDa protein alone can enhance rise. However, the increase in sensory evaluation value from the control for sample 2-2 (0.21) is greater than the predicted additive effect (0.17) of adding 2'DA and 40 kDa protein, calculated by adding the increase in sensory evaluation value from the control for sample 2-1 (0.11) and the increase in sensory evaluation value from the control for comparison sample 2-1 (0.06). It can be said that the combined use of 2'DA and 40 kDa protein exerts an unexpected synergistic effect in enhancing rise.

[0066] Example 3: Evaluation of the synergistic effect of 2'DA and 40 kDa protein 2'DA and 40 kDa protein were added to a commercially available beer-taste alcoholic beverage (referred to as beer-taste alcoholic beverage (C)), and a sensory evaluation was conducted. The beer-taste alcoholic beverage (C) evaluated in Example 3 is a beer-taste alcoholic beverage containing 50% or more by weight of malt as an ingredient. This beer-taste alcoholic beverage (C) is a different beer-taste alcoholic beverage from the commercially available beer-taste alcoholic beverage (A) used in Examples 1 and 2. The ingredients of the beer-flavored alcoholic beverage (C) are malt, hops, rice, corn, and starch, and its nutritional components per 100ml are 5% alcohol, 0.2-0.4g protein, 3.0g carbohydrates, and 5-6mg purines.

[0067] The concentrations of 2'DA and 40 kDa protein in the beer-flavored alcoholic beverage (C) were analyzed using the methods described above. The 2'DA concentration in the beer-flavored alcoholic beverage (C) (Control 2) was 10 ppm, and the 40 kDa protein concentration was 6.5 ppm. For the beer - flavored alcoholic beverage (C), 2’ - deoxyadenosine (2’DA) was added to a concentration of 11 ppm, and a sensory evaluation was conducted (Sample 3 - 2). Also, for the beer - flavored alcoholic beverage (C), 2’DA was added to a concentration of 11 ppm, and further, a 40 kDa protein was added to a concentration of 10 ppm, and a sensory evaluation was conducted (Sample 3 - 3).

[0068] The reference points and procedures for the sensory evaluation are the same as those in Example 1. The results of the sensory evaluation are shown in Table 3.

[0069]

Table 3

[0070] 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, adding 2’DA to a commercially available beer - flavored alcoholic beverage (C) with a malt ratio of 50 wt% or more in the raw materials can enhance the fullness. Furthermore, it can be seen that when 2’DA is 11 ppm and the 40 kDa protein concentration is 10 ppm, the fullness of the beer - flavored alcoholic beverage can be enhanced more effectively.

[0071] <II. Sensory evaluation when 2’DA is added to a commercially available beer - flavored alcoholic beverage (B) with a malt ratio of less than 50 wt% in the raw materials> 2’ - Deoxyadenosine (2’DA) was added to a commercially available beer - flavored alcoholic beverage (B), and a sensory evaluation of fullness was conducted. The beer - flavored alcoholic beverage (B) contains malt in the raw materials and has a malt ratio of less than 50 wt% in the raw materials. The raw materials are sparkling wine, malt, hops, sugars, dietary fiber, spirits (wheat), and contain 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.

[0072] (Analysis of commercially available beer-flavored alcoholic beverages) The concentration of 2'DA in the commercially available beer-flavored alcoholic beverage (B) used in the sensory evaluation was quantified by LC-MS using the same procedure as above. The concentration of 2'DA in the commercially available beer-flavored alcoholic beverage (B) was designated Control 3.

[0073] (Example 4: Evaluation by adding 2'DA) The concentration of 2'DA contained in the commercially available beer-flavored alcoholic beverage (B) (Control 3) was 1.9 ppm. To this, 2'DA was added so that the 2'DA concentrations were 9.2 ppm, 9.5 ppm, 15 ppm, 20 ppm, 50 ppm, and 100 ppm, and a sensory evaluation was performed. The criteria and procedures for the sensory evaluation were the same as those in Example 1. The results of the sensory evaluation are shown in Table 4.

[0074] [Table 4]

[0075] The results shown in Table 4 indicate that in beer-taste alcoholic beverages in which the proportion of malt in the raw materials is less than 50% by weight, fullness is enhanced when the 2'DA concentration is 9.2 ppm or higher.

[0076] Example 5: Evaluation of the synergistic effect of 2'DA and 40 kDa protein The synergistic effect of 2'DA and 40kDa protein was evaluated by adding 40kDa protein to a commercially available beer-flavored alcoholic beverage (B) (Control 3) containing 2'DA at a concentration of 9.2 ppm. The 40kDa protein concentrations were 1 ppm, 5 ppm, 10 ppm, and 25 ppm. The 40kDa protein used was purified as described above. For comparison, a commercially available beer-flavored alcoholic beverage (B) was also evaluated with the addition of only the 40 kDa protein. The results of the sensory evaluation are shown in Table 5.

[0077] [Table 5]

[0078] The results shown in Table 5 indicate that the swelling can be further enhanced by adding 2'DA and further adding 40 kDa protein to a commercially available beer-flavored alcoholic beverage (B) whose raw material contains less than 50% malt by weight. The results for comparison samples 5-1 to 5-3 demonstrate that the addition of 40 kDa protein alone can enhance swelling. However, the sensory evaluation increase (0.21) for sample 5-2 over the control is greater than the predicted additive effect (0.13) of combining 2'DA and 40 kDa protein, calculated by adding the sensory evaluation increase (0.10) for sample 5-1 and the sensory evaluation increase (0.03) for comparison sample 5-1. The sensory evaluation increase (0.35) for sample 5-3 over the control is also greater than the sum of the sensory evaluation increases (0.10 and 0.09) for sample 5-1 and comparison sample 5-2, respectively. The sensory evaluation increase (0.47) for sample 5-5 over the control is also greater than the sum of the sensory evaluation increases (0.10 and 0.20) for sample 5-1 and comparison sample 5-3, respectively. These results suggest that the combined use of 2'DA and 40kDa proteins produces an unexpected synergistic effect in enhancing the fullness of beer-flavored beverages containing less than 50% malt by weight of the raw materials. [Industrial Applicability]

[0079] According to the present invention, it is possible to provide a beer-taste alcoholic beverage with enhanced fullness, in which the malt content of the ingredients is 50% by weight or more. Also, according to the present invention, it is possible to provide a beer-taste alcoholic beverage with enhanced fullness, in which the malt content of the ingredients is less than 50% by weight.

Claims

1. A beer-flavored alcoholic beverage in which the ratio of malt in the raw materials is 50% by weight or more, and in which the concentration of 2'-deoxyadenosine is 11 ppm or more.

2. 2. The beer-taste alcoholic beverage according to claim 1, wherein the concentration of 2'-deoxyadenosine is 11 to 50 ppm.

3. Further, it contains a protein with a molecular weight of 35 to 50 kDa, 3. The beer-taste alcoholic beverage according to claim 1, wherein the protein concentration is 10 ppm or more.

4. 4. The beer-taste alcoholic beverage according to claim 3, wherein the protein concentration is 200 ppm or less.

5. A beer-flavored alcoholic beverage in which the proportion of malt in the raw materials is less than 50% by weight, and in which the concentration of 2'-deoxyadenosine is 9.2 ppm or more.

6. 6. The beer-taste alcoholic beverage according to claim 5, wherein the concentration of 2'-deoxyadenosine is 9.2 to 100 ppm.

7. Further, it contains a protein with a molecular weight of 35 to 50 kDa, 7. The beer-taste alcoholic beverage according to claim 5, wherein the protein concentration is 1 ppm or more.

8. 8. The beer-taste alcoholic beverage according to claim 7, wherein the protein concentration is 30 ppm or less.

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