Beer-taste alcoholic beverage

Abstract

The purpose of the present invention is to provide a beverage having an enriched bulging taste, in particular, a beer-taste alcoholic beverage which has an enriched bulging taste and has a purine concentration of 5 mg / 100 mL or more. The present invention pertains to a beer-taste alcoholic beverage which has a purine concentration of 5 mg / 100 mL or more and an adenosine concentration of 50 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 with 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 10-20 kDa at a predetermined concentration.

[0006] Here, as a taste that can be felt immediately after drinking a beer-taste beverage, there is a taste that cannot be expressed by the five basic tastes, namely, sweetness, saltiness, sourness, bitterness, and umami, and a feature such that the intensity, spread, thickness, duration of the taste, or the balance of the taste intensity is favorable may be preferred. Such a feature is 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 fizz. In particular, it aims to provide a beer - flavored alcoholic beverage with a concentration of purine bodies with enhanced fizz of 5 mg / 100 mL or more.

Means for Solving the Problems

[0008] That is, the present invention relates to the following beer - flavored alcoholic beverages. 〔1〕A beer - flavored alcoholic beverage in which the concentration of purine bodies is 5 mg / 100 mL or more and the concentration of adenosine is 50 ppm or more. 〔2〕The beer - flavored alcoholic beverage according to the above 〔1〕, in which the concentration of adenosine is 50 to 150 ppm. 〔3〕Furthermore, the beer - flavored alcoholic beverage according to the above 〔1〕 or 〔2〕, which contains a protein with a molecular weight of 35 to 50 kDa and the concentration of the above protein is 10 ppm or more. 〔4〕The beer - flavored alcoholic beverage according to the above 〔3〕, in which the concentration of the above protein is 200 ppm or less. 〔5〕A beer - flavored alcoholic beverage containing adenosine and a protein with a molecular weight of 35 to 50 kDa, in which the concentration of the above adenosine is 45 ppm or more, the concentration of the above protein is 30 ppm or more, and the concentration of purine bodies is 5 mg / 100 mL or more.

Effects of the Invention

[0009] According to the present invention, a beer - flavored alcoholic beverage with enhanced fizz can be provided.

Modes for Carrying Out the Invention

[0010] The beer-taste alcoholic beverage of the present invention has a purine body concentration of 5 mg / 100 mL or more. In this specification, the "purine body" is not particularly limited as long as it is a compound having a purine nucleus structure. Therefore, examples of the "purine body" include purine bases (adenine, guanine, xanthine, hypoxanthine), purine nucleosides (adenosine, guanosine, inosine), purine nucleotides (adenylic acid, guanylic acid, inosinic acid), and low-molecular or high-molecular nucleic acids (oligonucleotides, polynucleotides). In addition, in this specification, the "purine body concentration" refers to the total amount of the four purine body bases of adenine, guanine, xanthine, and hypoxanthine. The measurement of the purine body can be performed by a known method. For example, it can be measured by a method of detecting using LC-MS / MS (liquid chromatography-mass spectrometry) after hydrolysis with perchloric acid (see "Guidance on Trace Analysis of Purine Bodies in Liquors", Japan Food Analysis Center, URL: http: / / www.jfrl.or.jp / item / nutrition / post-31.html).

[0011] The concentration of the purine body in the beer-taste alcoholic beverage of the present invention is preferably 5.5 mg / 100 mL or more. Also, the concentration of the purine body in the beer-taste alcoholic beverage of the present invention is preferably 15 mg / 100 mL or less, and more preferably 13 mg / 100 mL or less.

[0012] The beer-taste alcoholic beverage of the present invention contains adenosine. Adenosine is a kind of nucleoside composed of adenine and ribose. Note that the above adenosine does not include derivatives of adenosine such as 5'-deoxyadenosine.

[0013] In the beer-taste alcoholic beverage of the present invention, the concentration of adenosine is 50 ppm or more. When the concentration of adenosine is 50 ppm or more, the swelling in a beer - flavored alcoholic beverage with a purine body concentration of 5 mg / 100 mL or more can be effectively enhanced. The fact that the presence of adenosine at a predetermined concentration or more is related to the swelling in a beer - flavored alcoholic beverage with a purine body concentration of 5 mg / 100 mL or more has not been known so far and is a finding discovered by the present inventors.

[0014] Moreover, in the beer - flavored alcoholic beverage of the present invention, the ratio of malt in the raw materials is preferably 50% by weight or more, and most preferably 100% by weight. Here, the "ratio of malt" refers to the ratio of the weight of malt in raw materials other than water and hops, such as malt, rice, corn, sorghum, potatoes, 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.

[0015] In the beer - flavored alcoholic beverage of the present invention, the concentration of adenosine is preferably 150 ppm or less. This is because if the concentration of adenosine becomes too high, the taste balance may be felt to be poor. In one aspect, the concentration of adenosine in the beer - flavored alcoholic beverage is preferably 50 - 150 ppm.

[0016] Moreover, the beer - flavored alcoholic beverage of the present invention further contains a protein with a molecular weight of 35 - 50 kDa, and the concentration of the above - mentioned protein is preferably 10 ppm or more. This is because when the protein concentration with a molecular weight of 35 - 50 kDa is 10 ppm or more, the swelling in a beer - flavored alcoholic beverage with a purine body concentration of 5 mg / 100 mL or more can be more effectively enhanced.

[0017] 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.

[0018] The 40 kDa protein is preferably a protein derived from cereal. The above cereal is preferably at least one selected from the group consisting of barley, wheat, corn, rice, and soybean. When the cereal is wheat, it can contain proteins derived from known wheat used in the production of beer-taste alcoholic beverages. Examples of such wheat include barley, wheat, rye, crow wheat, oat, and emmer wheat, preferably barley. Also, it can be either germinated wheat or ungerminated wheat, preferably malt of germinated wheat. These can be contained alone or in combination of two or more.

[0019] 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 its amino acid sequence are deleted, substituted, inserted, and / or added may be used.

[0020] By further containing the 40 kDa protein in addition to adenosine, the foaming in beer-taste alcoholic beverages can be effectively enhanced. Furthermore, the concentration of the 40 kDa protein is more preferably 25 ppm or more, even more preferably 30 ppm or more, and preferably 200 ppm or less.

[0021] Moreover, when both adenosine and the 40 kDa protein are contained, the synergistic effect of adenosine and the 40 kDa protein can enhance the foaming of a beer-taste alcoholic beverage with a purine concentration of 5 mg / 100 mL or more. Therefore, the lower limit of the concentration of adenosine required to enhance foaming may be low, and the lower limit of the concentration of the 40 kDa protein may also be low.

[0022] That is, another form of the beer-taste alcoholic beverage of the present invention contains adenosine and a protein having a molecular weight of 35 to 50 kDa, the concentration of the adenosine is 45 ppm or more, the concentration of the protein is 30 ppm or more, and it is a beer-taste alcoholic beverage with a purine concentration of 5 mg / 100 mL or more.

[0023] 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 fermentation or non-fermentation. Here, the alcohol refers to ethanol and does not include the aliphatic alcohols described later. A beer-taste beverage refers to a carbonated beverage having a beer-like flavor.

[0024] The manufacturing process of a general beer-taste alcoholic beverage is shown below. First, to a mixture containing, in addition to wheat such as malt, other grains, starch, sugars, bittering agents, coloring agents, etc. as required, and water, enzymes such as amylase are added as required, followed by gelatinization and saccharification, and then filtration to obtain a saccharified liquid. As required, 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, hops may be added to a mixture of malt extract and warm water 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 beer storage processes, etc. may be those known conditions. The obtained fermented liquid is filtered, and carbon dioxide gas is added to the obtained filtrate. Thereafter, it is filled into a container and passed through a sterilization process to obtain the target beer-taste alcoholic beverage. The beer-taste alcoholic beverage may be produced by adding an alcohol beverage derived from wheat, such as a wheat-derived distilled liquor (e.g., spirits, shochu, etc.), to the beer-taste alcoholic beverage produced by the method as described above.

[0025] The beer-taste alcoholic beverage produced without using malt is prepared by mixing liquid sugar containing a carbon source, a nitrogen source as an amino acid-containing material other than wheat or malt, hops, pigments, etc. with warm water to obtain 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 to a mixture of an extract using raw materials other than malt and warm water and boiled. Hops may be mixed at any stage from the start of boiling to before the end of boiling. Conditions in the fermentation and beer storage processes, etc. may be those known conditions. The obtained fermented liquid is filtered, and carbon dioxide gas is added to the obtained filtrate. Thereafter, it is filled into a container and passed through a sterilization process to obtain the target beer-taste alcoholic beverage.

[0026] The non-fermented and alcohol-containing beer-taste alcoholic beverage may be one in which the alcohol content of the final product is adjusted by adding raw material alcohol or the like. The addition of raw material alcohol may be carried out at any step from the saccharification step to the filling step.

[0027] 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 from the beverage is prepared, and the sample is distilled over 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 annex to the prescribed 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.

[0028] From the viewpoint of imparting a liquor flavor, 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 aliphatic alcohols having 4 to 5 carbon atoms are preferred. In the present invention, preferred aliphatic alcohols having 4 carbon atoms include 2-methyl-1-propanol, 1-butanol, etc., and those having 5 carbon atoms include 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 using the headspace gas chromatography method.

[0029] 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 the 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.

[0030] 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.

[0031] In the beer-taste alcoholic beverage according to the present invention, hop can be used as 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.

[0032] The beer-taste alcoholic beverage according to the present invention may, within a range not interfering with the effects of the present invention, optionally use other raw materials as necessary. For example, sweeteners (including high-intensity sweeteners), bittering agents, flavorings, yeast extracts, coloring agents such as caramel pigments, plant-extracted saponin substances such as soybean saponin and quillaja saponin, plant protein and peptide-containing substances such as corn and soybean, 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 a range not interfering with the effects of the present invention.

[0033] The beer-taste alcoholic beverage according to the present invention can be packaged in a container. 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.

[0034] The method for producing the beer-taste alcoholic beverage of the present invention is not particularly limited, but a method of adding a predetermined amount of adenosine to a beer-taste alcoholic beverage having a purine concentration of 5 mg / 100 mL or more is exemplified. Moreover, it is preferable to add a 40 kDa protein to a beer-taste alcoholic beverage having a purine concentration of 5 mg / 100 mL or more. The preparation of the adenosine and 40 kDa protein to be added can be carried out, for example, according to the procedure described in the examples below. Moreover, regarding adenosine and 40 kDa protein, their contents may be increased by adjusting various conditions in the production process of the beer-taste alcoholic beverage.

Examples

[0035] Hereinafter, the present invention will be specifically described by showing examples, but the present invention is not limited to the following examples.

[0036] (Purification of Adenosine) The adenosine was purified according to the following procedure. (1) Fractionation of beer by HP20 60 L of beer was fractionated using 10 L of Diaion (registered trademark) 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 large-scale fractionation column and replaced with water. The same amount of distilled water was mixed with 60 L of degassed beer and flowed into the HP20 column using a medium-pressure pump. The solution that passed through the HP20 column was obtained as the through-fraction. 40 L of distilled water was flowed 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, 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 dry product using an evaporator and a freeze dryer.

[0037] (2) Fractionation of the 10% ethanol elution fraction by LH-20 Among the HP20 fractions, the 10% ethanol elution fraction was fractionated using Sephadex (registered trademark) LH-20 (column dosage: 500 mL). LH-20 washed with ethanol was packed into a large-scale fractionation column and replaced with water. The 10% ethanol elution fraction (2.57 g) obtained by HP20 fractionation was dissolved in distilled water and applied to the LH-20 column. 1 L of distilled water was flowed to obtain water elution fractions -1 to 5. Next, 1 L each of aqueous ethanol (35% ethanol, 70% ethanol, and 100% ethanol) was flowed, 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 dry product using an evaporator and a freeze dryer.

[0038] (3) Separation of adenosine For the water elution fraction - 3 (82.4 mg) obtained by LH - 20 fractionation, HPLC (COSMOSIL 5C18 - PAQ, 20×250 mm) was used and eluted with 10% ethanol. Then, the eluate from 5 min to 12 min was concentrated and eluted with a mixed solution of ethanol - water (5:95 → 15:85) concentration gradient using HPLC (COSMOSIL 5C18 - PAQ, 20×250 mm) to obtain compound (I) (tR = 18.5 min). Compound (I) was identified as adenosine by analysis of physical data of MS and NMR and comparison with the reference standard. The analytical instruments used are as follows. LC - MS; Q Exactive, manufactured by Thermo Fisher Scientific NMR; AVANCE400, manufactured by Bruker

[0039] (Purification of 40 kDa protein) Purification of 40 kDa protein was carried out from commercially available beer (1 L) according to the following procedure.

[0040] (1) Fractionation by cation - exchange resin 50 mL of cation - exchange resin SP Sepharose was placed in an empty column. Beer was adsorbed onto the resin. Then, the resin was transferred to the column and washed with 20 mM sodium acetate buffer (pH 4.5). Next, it was eluted 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.

[0041] (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.

[0042] (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 40 kDa protein purified product (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.

[0043] 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 digest was recovered. The recovered solution was dried under reduced pressure and redissolved in a 0.1% formic acid solution. This was used for LC-MS / MS analysis.

[0044] (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 (Nikkaki Biosciences Co., Ltd.) Liquid chromatograph 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

[0045] (4) Protein Analysis 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 Miss Cleavage Number: 5 Confidence Level (Percolator): High (the highest probability level among the three levels of certainty) Database: SwissProt

[0046] 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%).

[0047] (Sensory Evaluation When Adenosine was Added to Commercial Beer - Taste Alcoholic Beverage 1) Adenosine was added to commercial beer - taste alcoholic beverage 1, and a sensory evaluation of the fullness was conducted. The beer-taste alcoholic beverage is a beer-taste alcoholic beverage in which the ratio of malt in the raw materials is 50% by weight or more. The raw materials of the beer-taste 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.

[0048] The criteria points for 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 swelling intensity is based on the following criteria. 0 points: Not felt at all 1 point: Slightly felt 2 points: Clearly felt 3 points: Felt very strongly As a reference point, the swelling of the same beer-taste alcoholic beverage as the commercially available beer-taste alcoholic beverage 1 to be evaluated was set at 1.5 points as the reference beer-taste alcoholic beverage (I). Also, for the same beer-taste alcoholic beverage as the commercially available beer-taste alcoholic beverage 1 to be evaluated, the swelling of the beverage with 25 ppm of 40 kDa protein added to make the 40 kDa protein concentration 50 ppm was set at 2 points.

[0049] The procedure for sensory evaluation is as follows. (1) Dispense the beer-taste alcoholic beverage into a vial at 1 / 10 of the final volume (v / v). (2) Weigh adenosine in an arbitrary amount and add it. (3) Sonicate for 30 seconds. (4) Let it stand at room temperature for 30 minutes. (5) Fill up the beer-taste alcoholic beverage to the final volume. (6) Dispense and drink for evaluation.

[0050] (Analysis of Commercially Available Beer-Taste Alcoholic Beverages) The concentration of adenosine contained in a 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 construction of calibration curve For adenosine, 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) aqueous ethanol solution was used as the diluent. In addition, in the analysis results of the standard, the measured values of the dilution ratio were adopted such that the measured values fell within the range where the linearity of the calibration curve was maintained (R 2 > 0.99).

[0051] 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

[0052] (2) Preparation of measurement sample from commercially available beer-taste alcoholic beverage A commercially available beer - flavored alcoholic beverage was degassed by sonication. After the bubbles settled, 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 adenosine in commercially available beer - flavored alcoholic beverage 1 was taken as control 1.

[0053] The concentration of the 40 - kDa protein contained in commercially available beer - flavored alcoholic beverage 1 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 contained in commercially available beer - flavored alcoholic beverage 1 was 25 ppm.

[0054] (Example 1: Evaluation by addition of adenosine) The concentration of adenosine in commercially available beer - flavored alcoholic beverage 1 (control 1) was 41.9 ppm. In contrast, adenosine was added so that the adenosine concentrations were 50 ppm, 100 ppm, and 150 ppm respectively, and sensory evaluation was carried out (sample 2, sample 4, and sample 5). The results of the sensory evaluation are shown in Table 1.

[0055] (Example 2: Evaluation of the synergistic effect of adenosine and 40 - kDa protein) To commercially available beer - flavored alcoholic beverage 1 (control 1), 40 - kDa protein and adenosine were added so that the 40 - kDa protein concentration was 30 ppm and the adenosine concentrations were 45 ppm and 50 ppm respectively, and a sensory evaluation test was carried out (sample 1 and sample 3). As the 40 - kDa protein, the one purified above was used. Also, for comparison, only 5 ppm of 40 - kDa protein was added to commercially available beer - flavored alcoholic beverage 1, and an evaluation was carried out when the 40 - kDa protein concentration was 30 ppm (comparative sample 1). The results of the sensory evaluation are shown in Table 1. In addition, the purine body concentrations of each sample are shown in Table 1 below. The "purine body concentration" in this specification is the total amount of four purine bases, namely adenine, guanine, xanthine, and hypoxanthine, as described above, and can be detected, for example, by LC-MS / MS (liquid chromatography-mass spectrometry) after hydrolysis with perchloric acid.

[0056] [Table 1]

[0057] From the results of Sample 2, Sample 4, and Sample 5 shown in Table 1, for beer-taste alcoholic beverages with a purine body concentration of 5 mg / 100 mL or more, when the adenosine concentration is 50 ppm or more, the average of the sensory evaluation is 0.1 point or more greater than that of the control (1.5), and it can be seen that the swelling is effectively enhanced. Thus, it can be said that by setting the concentration of adenosine in beer-taste alcoholic beverages with a purine body concentration of 5 mg / 100 mL or more to a predetermined concentration or more, the swelling can be effectively enhanced compared to the control. In addition, from the results of Comparative Sample 1, it can be seen that even by simply adding a 40 kDa protein to a beer-taste alcoholic beverage, it becomes 0.08 points greater than the control, and the swelling can be enhanced. However, the increase value from the control of the sensory evaluation in Sample 1 in which adenosine and a 40 kDa protein are used in combination is 0.1 point or more, and it can be understood that even when the adenosine concentration is less than 50 ppm, the swelling is effectively enhanced by being used in combination with a 40 kDa protein. Thus, it can be said that a synergistic effect that cannot be expected in enhancing the swelling is exhibited by using adenosine and a 40 kDa protein in combination.

[0058] (Example 3: Evaluation by Adenosine Addition) The concentration of adenosine contained in the commercially available beer - flavored alcoholic beverage 2 (Control 2) was 17.6 ppm. Also, the concentration of the 40 kDa protein was 6.5 ppm. Adenosine was added to this commercially available beer - flavored alcoholic beverage 2 so that the adenosine concentration became 50 ppm, and a sensory evaluation was conducted (Sample 6). Also, adenosine was added to the commercially available beer - flavored alcoholic beverage 2 so that the adenosine concentration became 50 ppm, and further, a 40 kDa protein was added so that the concentration of the 40 kDa protein became 10 ppm, and a sensory evaluation was conducted (Sample 7). The procedure of the sensory evaluation was the same as the procedure in Example 1, and the reference points in the sensory evaluation were as follows. The results of the sensory evaluation are shown in Table 2. Also, the purine - body concentrations of each sample are shown in Table 2 below. The "purine - body concentration" in this specification is the total amount of the four purine - body bases of adenine, guanine, xanthine, and hypoxanthine as described above, and can be detected, for example, using LC - MS / MS (liquid chromatography - mass spectrometry) after hydrolysis with perchloric acid.

[0059] Note that the beer - flavored alcoholic beverage 2 evaluated in Example 3 is a beer - flavored alcoholic beverage in which the ratio of malt in the raw materials is 50% by weight or more. The beer - flavored alcoholic beverage 2 is a different beer - flavored alcoholic beverage from the commercially available beer - flavored alcoholic beverage 1 used in Examples 1 and 2. The raw materials of the beer - flavored alcoholic beverage 2 are malt, hops, rice, corn, and starch, and it contains 5% alcohol, 0.2 - 0.4 g of protein, 3.0 g of carbohydrates, and 5 - 6 mg of purine - bodies per 100 ml as nutritional components.

[0060] The reference points and procedures of the sensory evaluation are the same as the reference points and procedures in Example 1. The results of the sensory evaluation are shown in Table 2.

[0061]

Table 2

[0062] From the results of Sample 6 shown in Table 2, for beer - flavored alcoholic beverages 2 with a purine body concentration of 5 mg / 100 mL or more, when the adenosine concentration is 50 ppm or more, even when the 40 kDa protein concentration is as low as 6.5 ppm, the average of the sensory evaluation becomes 0.07 points higher than that of Control 2 (1.0), and it can be seen that the swelling is enhanced. Thus, even when the 40 kDa protein concentration is low, by setting the adenosine concentration to a predetermined concentration or more, it can be said that the swelling can be enhanced compared to Control 2. Furthermore, the increase value from Control 2 in the sensory evaluation of Sample 7 where the adenosine concentration is 50 ppm and the 40 kDa protein is 10 ppm is 0.1 point or more. It can be understood that when the adenosine concentration is a predetermined concentration or more and the 40 kDa protein concentration is a predetermined concentration or more, the swelling is effectively enhanced. Thus, it can be said that when the adenosine concentration is 50 ppm or more and the 40 kDa protein concentration is 10 ppm or more, a synergistic effect that cannot be predicted in enhancing the swelling is exhibited.

Industrial Applicability

[0063] According to the present invention, in a beer - flavored alcoholic beverage having a purine body concentration of 5 mg / 100 mL or more, a beverage with enhanced swelling can be provided.

Claims

【Claim 1】 A beer-taste alcoholic beverage having a purine concentration of 6.98 to 17.97 mg / 100 mL, an adenosine concentration of 50 to 150 ppm, and a total concentration of barley-derived Serpin Z4 and barley-derived Serpin Z7 of 25 to 30 ppm.

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