TW202207814A - Beer-taste alcoholic beverage - Google Patents

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

beer-flavored alcoholic beverages

The present invention relates to a beer-flavored alcoholic beverage.

With the diversification of consumers' tastes in recent years, the development of a beer-flavored alcoholic beverage having various flavor characteristics is desired.

Patent Document 1 discloses a peptide containing a specific molecular weight in order to improve the flavor of a beer-flavored alcoholic beverage. [Prior Art Literature] [Patent Literature]

[Patent Document 1] Japanese Patent Laid-Open No. 2016-149975

[Problems to be Solved by the Invention]

Patent Document 1 describes a beverage, which is used as an evaluation index for the beverage, using such indicators as beer-like flavor, smooth taste flow, and gritty feeling remaining in the mouth. , the sensory evaluation score based on these indicators is higher.

Here, as a taste immediately after drinking a beer-flavored beverage, the five basic tastes, namely, sweet, salty, sour, bitter, and umami, cannot be expressed as the intensity, diffusion, and richness of the taste. Characteristics of a balance of intensity, flavor persistence or flavor intensity are preferred. Such a feature is called "swelling degree" in this specification. The beverage described in Patent Document 1 is a beverage with room for improvement from the viewpoint of the swelling degree, and a method for enhancing the swelling degree of a beer-flavored alcoholic beverage is desired.

The purpose of the present invention is to provide a beverage with enhanced swelling. In particular, the purpose is to provide a beer-flavored alcoholic beverage with a purine body concentration of 5 mg/100 mL or more with an enhanced swelling degree. [Means of Solving Problems]

That is, the present invention relates to one of the following beer-flavored alcoholic beverages. [1] A beer-flavored alcoholic beverage, wherein the concentration of purine bodies is 5 mg/100 mL or more, and the concentration of nucleosides is 50 ppm or more. [2] The beer-flavored alcoholic beverage according to the above [1], wherein the concentration of adenosine is 50-150 ppm. [3] The beer-flavored alcoholic beverage according to the above [1] or [2], further comprising 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-flavored alcoholic beverage according to the above [3], wherein the concentration of the protein is 200 ppm or less. [5] A beer-flavored alcoholic beverage with a purine concentration of 5 mg/100 mL or more, comprising adenosine and a protein with a molecular weight of 35 to 50 kDa, wherein the adenosine concentration is 45 ppm or more, and the protein concentration is 30 ppm above. [Effect of invention]

According to the present invention, a beer-flavored alcoholic beverage with an enhanced swelling degree can be provided.

The concentration of purine bodies in the beer-flavored alcoholic beverage of the present invention is 5 mg/100 mL or more. In this specification, "purine body" means a compound as long as it has a purine core structure, and is not particularly limited. Therefore, examples of "purinosomes" include purine bases (adenine, guanine, xanthine, hypoxanthine), purine nucleosides (adenosine, guanosine, inosine), and purine nucleotides (adenosine). Nucleotides, bird nucleotides, inosinic acid) and low-molecular or high-molecular nucleic acids (oligonucleotides, polynucleotides), etc. In addition, in this specification, the "purinosome concentration" means the total amount of 4 types of purinosome bases of adenine, guanine, xanthine, and hypoxanthine. The determination of purine bodies can be carried out by a known method, for example, by a method that can be measured by using LC-MS/MS (liquid chromatography mass spectrometry) after hydrolysis with perchloric acid (“Purine bodies in wines”). "Introduction to Microanalysis", General Foundation, Japan Food Analysis Center, URL: http://www.jfrl.or.jp/item/ nutrition/post-31.html) to measure.

The concentration of purine bodies in the beer-flavored alcoholic beverage of the present invention is preferably 5.5 mg/100 mL or more. Furthermore, the concentration of purine bodies in the beer-flavored alcoholic beverage of the present invention is preferably 15 mg/100 mL or less, and even more preferably 13 mg/100 mL or less.

Adenosine is contained in the beer-flavored alcoholic beverage of the present invention. Adenosine is a type of nucleosides containing adenine and ribose. In addition, the above-mentioned adenosine is not a derivative of adenosine including 5'deoxyadenosine.

In the beer-flavored 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 degree of a beer-flavored alcoholic beverage having a purine concentration of 5 mg/100 mL or more can be effectively enhanced. The fact that the present inventors discovered that the swelling degree of a beer-flavored alcoholic beverage containing a specific concentration or more of adenosine and a purine concentration of 5 mg/100 mL or more has not been known until now.

Furthermore, the ratio of malt in the raw material of the beer-flavored alcoholic beverage of the present invention is preferably 50% by weight or more, and most preferably 100% by weight. Here, the "ratio of malt" means the ratio of the weight of malt, such as malt, rice, corn, sorghum, potato, starch, wheat other than malt, and sugar, to water and malt other than hops. However, ingredients that can be added in trace amounts, such as sour, sweet, bitter, seasoning, and spice, are not included in the calculation of the above ratio.

In the beer-flavored alcoholic beverage of the present invention, the concentration of adenosine is preferably 150 ppm or less. This is because when the concentration of adenosine becomes too high, a poor balance of taste may be felt. In one form, the concentration of adenosine in the beer-flavored alcoholic beverage is preferably 50-150 ppm.

In addition, the beer-flavored alcoholic beverage of the present invention further contains a protein with a molecular weight of 35 to 50 kDa, and the concentration of the protein is preferably 10 ppm or more. If the concentration of protein with a molecular weight of 35 to 50 kDa is 10 ppm or more, the swelling degree of beer-flavored alcoholic beverages with a purine concentration of 5 mg/100 mL or more can be more effectively enhanced.

A protein with a molecular weight of 35 to 50 kDa is a protein whose molecular weight appears in the region of 35 to 50 kDa when a beer-flavored alcoholic beverage is subjected to SDS-PAGE electrophoresis. Before supplying the beer-flavored alcoholic beverage to the electrophoresis of SDS-PAGE, for example, the beer-flavored alcoholic beverage may be subjected to extra-limited filtration using a 30 kDa cut off membrane as a pretreatment. The above-mentioned protein is preferably a protein with a molecular weight of 35-45 kDa, more preferably a protein of about 40 kDa. In this specification, proteins with a molecular weight of 35 to 50 kDa are also referred to as 40 kDa proteins.

The 40kDa protein is preferably a protein derived from cereals. It is preferable that the above-mentioned cereals are at least one selected from the group consisting of barley, wheat, corn, rice, and soybeans. Moreover, when a cereal grain is wheat, the protein derived from the well-known wheat used for manufacture of a beer-flavored alcoholic beverage can be contained. Examples of such wheat include barley, wheat, rye, black wheat, barley, oats, and the like, and barley is preferred. In addition, it may be either malted wheat or unmalted wheat, but it is preferably malted wheat malt. These may be contained individually or in combination of 2 or more types.

Further, as the 40 kDa protein, Serpin Z4 (alias: BSZ4, HorvuZ4, Major endosperm albumin or Protein Z) derived from barley (scientific name: Hordeum vulgare) and Serpin Z7 (alias: BSZ7 or HorvuZ7) derived from barley are preferable. In addition, among the above-mentioned proteins, amino acids which are part of the amino acid sequence may also have amino acid sequences deleted, substituted, inserted and/or added.

In addition to adenosine, the swelling degree of beer-flavored alcoholic beverages can be effectively enhanced by further containing 40 kDa protein. Furthermore, the concentration of the 40 kDa protein is more preferably 25 ppm or more, more preferably 30 ppm or more, and more preferably 200 ppm or less.

Furthermore, when both adenosine and 40kDa protein are contained, the swelling degree of a beer-flavored alcoholic beverage with a purine concentration of 5mg/100mL or more can be enhanced by the synergistic effect of adenosine and 40kDa protein. Therefore, the lower limit of the concentration of adenosine necessary for enhancing the swelling degree can be lower, and the lower limit of the concentration of 40kDa protein can also be lower.

That is, another form of the beer-flavored alcoholic beverage of the present invention is a beer-flavored alcoholic beverage with a purine concentration of 5 mg/100 mL or more, which comprises adenosine and a protein with a molecular weight of 35 to 50 kDa, and the The concentration is 45 ppm or more and the concentration of the above-mentioned protein is 30 ppm or more.

The beer-flavored alcoholic beverage of the present invention is a beer-flavored beverage containing alcohol, and the alcohol concentration is preferably 1% (v/v) to 10% (v/v), but is not particularly limited. Furthermore, the source of the alcohol component contained in the beer-flavored alcoholic beverage is not limited to fermentation and non-fermentation. In addition, the alcohol here means ethanol, and is not included in the aliphatic alcohol mentioned later. The beer-flavored beverage means a carbonated beverage having a beer-like flavor.

The manufacturing steps of general beer-flavored alcoholic beverages are shown below. First of all, in addition to the wheat including malt, other raw materials such as grains, starches, sugars, bitters or colorants and water as necessary, enzymes such as amylase are added as necessary to perform gelatinization, Saccharified and filtered as saccharified liquid. If necessary, add hops, bitters, etc. to the saccharification solution, boil it, and remove solids such as coagulated proteins in a clarification tank. As an alternative to this saccharification solution, hops may be added to the malt extract with warm water added and boiled. Hops can also be blended at any stage from the start of boiling until the end of boiling. The conditions in the saccharification step, boiling step, solid content removal step, etc. may be well-known conditions. For conditions such as fermentation and wine storage steps, well-known conditions should be used. The obtained fermentation broth was filtered, and carbon dioxide gas was added to the obtained filtrate. Then, it is filled in a container, and it goes through a sterilization process to obtain the desired beer-flavored alcoholic beverage. The beer-flavored alcoholic beverage may be produced by adding wheat-derived alcoholic beverages such as barley-derived distilled spirits (eg, distilled spirits, shochu, etc.) to the beer-flavored alcoholic beverages produced by the above-described method.

Beer-flavored alcoholic beverages manufactured without using malt as a raw material are liquid sugar containing carbon sources, nitrogen sources other than malt or malt as amino acid-containing materials, hops, pigments, etc. are mixed with warm water, as Liquid sugar solution. The liquid sugar solution is boiled. When using hops as a raw material, the hops may be mixed with the liquid sugar solution during the boiling, not before the start of boiling. As an alternative to this saccharified liquid, it is also possible to add hops to an extract using a raw material other than malt and to add warm water, and then boil it. Hops can also be blended at any stage from the start of boiling until the end of boiling. For conditions such as fermentation and wine storage steps, well-known conditions should be used. The obtained fermentation broth was filtered, and carbon dioxide gas was added to the obtained filtrate. Then, it is filled in a container, and it goes through a sterilization process to obtain the desired beer-flavored alcoholic beverage.

For non-fermented beer-flavored alcoholic beverages containing alcohol, the alcohol content of the final product can also be adjusted by adding alcohol for raw materials. The addition of the alcohol for raw materials can also be carried out in any step from the saccharification step to the filling step.

The alcohol content of the beer-flavored alcoholic beverage according to the present invention means the content (v/v%) of the alcohol component in the beverage, and can be measured by any known method, but can be measured by, for example, a vibrating density meter. Specifically, it is possible to prepare a sample for removing carbonic acid gas from beverages by filtration or ultrasonic waves, and subject the sample to direct-fire distillation to measure the density of the obtained distillate at 15°C, using the analytical method specified by the National Taxation Bureau (flat). 19. Order No. 6 of the National Taxation Bureau (revised on June 22, 2019) in the attached table "Table 2 Alcohol Composition and Density (15°C) and Specific Gravity (15/15°C) Conversion Table" to convert and calculate. When the alcohol content is less than 1.0%, a commercially available alcohol measuring device or gas chromatography can also be used.

In the beer-flavored alcoholic beverage according to the present invention, an aliphatic alcohol may be added from the viewpoint of imparting a sense of alcohol. 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, as preferable aliphatic alcohols, those having 4 carbon atoms include 2-methyl-1-propanol, 1-butanol, and the like, 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 the present specification, the content of the aliphatic alcohol can be measured using headspace gas chromatography.

The saccharide contained in the beer-flavored alcoholic beverage according to the present invention means the saccharide based on the nutrition representation standard for foods (Ministry of Health, Labour and Welfare Notice No. 176 in 2005). Specifically, a carbohydrate means a protein, lipid, dietary fiber, ash, alcohol content, and water that are removed from food. Also, the amount of sugar in a food can be calculated by deducting the amounts of protein, lipid, dietary fiber, ash, and moisture from the weight of the food. At this time, the amounts of protein, lipid, dietary fiber, ash, and moisture were measured by the methods disclosed in the Nutritional Standard. Specifically, the amount of protein was determined by nitrogen quantitative conversion method, the amount of lipid was determined by ether extraction method, chloroform-methanol mixed liquid extraction method, Gerber method, acid decomposition method or Reese-Gottlieb method, dietary fiber The amount of ash is determined by high-speed liquid chromatography or Prosky method, the amount of ash is determined by magnesium acetate adding ashing method, direct ashing method or sulfuric acid adding ashing method, and the amount of moisture is determined by Karl-Fischer method, drying aid method. Dosage method, reduced pressure superheat drying method, atmospheric pressure heating drying method or plastic film method to measure.

The beer-flavored alcoholic beverage related to the present invention caters to the preference for low sugar content in recent years, and can also be low sugar. Therefore, the sugar content of the beer-flavored alcoholic beverage related to the present invention may also be less than 2.5 g/100 mL or less than 0.5 g/100 mL. In addition, the lower limit is not particularly set, but is usually about 0.1 g/100 mL, for example, 0.15 g/100 mL or more or 0.2 g/100 mL or more.

In the beer-flavored alcoholic beverage according to the present invention, hops can be used as part of the raw material. When hops are used, general flake hops, powdered hops, and hop extracts used in the production of beer and the like can be appropriately selected and used according to the desired flavor. In addition, processed hops such as dissimilated hops and reduced hops can also be used. These are included in the hops used in the beer-flavored alcoholic beverage related to the present invention. In addition, the addition amount of hops is not particularly limited, but is typically about 0.0001 to 1% by weight with respect to the total amount of the beverage.

In the beer-flavored alcoholic beverage related to the present invention, other raw materials may be used as necessary within the range that does not hinder the effects of the present invention. Coloring materials such as sweeteners (including high-intensity sweeteners), bitters, flavors, yeast extracts, caramel colors, and soybean saponins can be used as necessary within the range that does not hinder the effects of the present invention. Or plant extract saponins such as saponins, plant proteins such as corn or soybeans and substances containing peptides, bovine serum albumin and other protein-based substances, dietary fibers or amino acids and other seasonings, ascorbic acid, etc. of antioxidants.

The beer-flavored alcoholic beverage related to the present invention can be packaged as a container. There is no restriction on the form of the container, and it can be filled in sealed containers such as bottles, cans, barrels, or pet bottles, and can be used as beverages contained in containers.

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

The present invention will be specifically described below with reference to examples, but the present invention is not limited to the following examples.

(purification of adenosine) Purification of adenosine was carried out as follows. (1) Fractionation of HP20 of beer 60 L of beer was fractionated using 10 L of Diaion (registered trademark) HP20 (manufactured by Mitsubishi Chemical Co., Ltd.). The HP20 was washed three times with ethanol and then three times with 50% ethanol before use. The washed HP20 was packed in a column for a large number of fractionation and replaced with water. Mix the degassed 60L beer with the same amount of distilled water, use a medium pressure pump, and flow into the HP20 column. A solution that passed through the HP20 column was obtained as the pass fraction. Using a medium-pressure pump, 40 L of distilled water was poured into the solution to obtain an elution solution, which was used as a water elution fraction. Similarly, 40 L of water-containing ethanol (10% ethanol, 30% ethanol, and 70% ethanol) were each poured to obtain eluates, which were used as 10% ethanol eluted fractions, 30% ethanol eluted fractions, and 70% ethanol eluted fractions. Each eluted fraction was refrigerated and stored as a dried product using an evaporator and a freeze dryer.

(2) LH-20 Fractionation of 10% Ethanol Dissolution Fraction Among the HP20 fractions, the fraction eluted with 10% ethanol was fractionated using Sephadex (registered trademark) LH-20 (column amount: 500 mL). The LH-20 washed with ethanol was packed into a column for fractionation in large quantities, and replaced with water. The 10% ethanol-eluted fraction (2.57 g) obtained by fractional distillation with HP20 was dissolved in distilled water and used for LH-20 column. Flow into 1L of distilled water to obtain water-dissolved fractions -1~5. Then, 1 L of water-containing ethanol (35% ethanol, 70% ethanol and 100% ethanol) was respectively poured into the solution to obtain eluates, which were respectively used as 35% ethanol eluted fraction, 70% ethanol eluted fraction and 100% ethanol eluted fraction. The respective eluted fractions were refrigerated and stored as dried products using an evaporator and a freeze dryer.

(3) Separation of adenosine The water-eluted fraction-3 (82.4 mg) obtained by fractional distillation of LH-20 was eluted with 10% ethanol using HPLC (COSMOSIL 5C18-PAQ, 20×250 mm). Next, the eluate was concentrated from 5 minutes to 12 minutes, and eluted with a mixture of a concentration gradient of ethanol-water (5:95→15:85) using HPLC (COSMOSIL 5C18-PAQ, 20×250 mm), Compound (I) was obtained (tR=18.5 min). Compound (I) was identified as adenosine by analysis of the physical data of MS and NMR and comparison with the standard substance. The analytical machine used is as described below. LC-MS; Q Exactive, manufactured by Thermo Fisher Scientific NMR; AVANCE400, manufactured by Bruker

(purification of 40kDa protein) Purification of the 40 kDa protein was performed from a commercial beer (1 L) as described below.

(1) Fractionation of cation exchange resin The cation exchange resin SP Sepharose 50mL was put into the empty column. Make the beer adsorb the resin. After that, the resin was converted into a 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 resulting fractions were evaluated by SDS-PAGE, and the fractions contained in the 40 kDa protein were collected as cation exchange resin bound fractions.

(2) Filter outside the limit (buffer exchange) 10 mL of each of the cation exchange resin-bound fractions obtained in (1) was added to an extra-limit filtration unit (Amicon Ultra-15 30K manufactured by Merck) after washing with water, and centrifuged at 3500 rpm to obtain an extra-limit filtration concentrate.

(3) Ammonium sulfate fractionation 20 mM phosphate buffer (pH 7.0) + 2 M ammonium sulfate was added to the beaker, and the concentrated solution obtained in (2) was added dropwise and stirred. The suspension was then centrifuged (2330 g, 10 min, room temperature). Collect the supernatant in another container. The collected solution was concentrated using an out-of-limit filter unit. The concentrate was added to 20 mM sodium acetate (pH 4.5), centrifuged (2330 g, 10 minutes, room temperature), and concentrated to obtain a 40 kDa protein purified product (Bradford quantification (bovine serum albumin (BSA) conversion), 20.4 mg/mL, 2.21 mL). The purity of the obtained 40 kDa protein was confirmed by SDS-PAGE.

After the 40kDa protein was digested with enzymes, the identification of the protein was attempted by analysis by LC-MS/MS. Fragments around 40kDa separated by SDS-PAGE were excised and subjected to reduction of dithiothreitol (56°C, 1 hour) and methylation of carbamoylamine with iodoacetamide (under light-shielding, room temperature, 45 minutes) ). Next, 15 μL of 10 ng/μL chymotrypsin solution (5 mM calcium chloride, 50 mM ammonium bicarbonate solution), 15 μL of 5 mM calcium chloride, 50 mM ammonium bicarbonate solution containing 0.01% ProteaseMax were added, and after overnight incubation, the enzyme digestion was recovered. liquid. The recovered solution was dried under reduced pressure, and then dissolved in 0.1% formic acid solution. This was used in LC-MS/MS analysis.

(Measurement by LC-MS/MS) The measurement by LC-MS/MS was carried out under the following conditions. Equipment used: direct flow nanoLC system Easy-nLC 1000TM (Thermo Scientific) Capture column: Acclaim PepMap (registered trademark) (Thermo Scientific) Analytical column: NANO HPLC CAPILLARY COLUMN (Nikyo-tec Co., Ltd.) Liquid chromatography mass analyzer Q Exactive Plus (Thermo Scientific) Mobile phase: solution A: 0.1% formic acid/water, solution B: 0.1% formic acid/acetonitrile Flow rate: 300nL/min Gradient: 0-40%B/0-30min, 40-60%B/30-35min, 60-90%B/35-37min, 90%B/37-45min Injection volume: 10 μL Ionization Mode: ESI Positive Measurement range: MS1 (m/z 350-1750) Data Dependent Scan mode

(4) Analysis of protein Protein identification was performed under the following conditions. Search software: Proteome Discoverer 2.2.0.388 (manufactured by ThermoFisher) Biological species: barley (Hordeum vulgare), hops (Humulus), yeast (Saccharomyces cerevisiae) Search terms: Digestive enzyme: chymotrypsin Precursor ion mass error range: monoisotope, ±10ppm Product ion mass error range: ±0.02Da Maximum number of missed cleavages: 5 Percolator: High (the highest degree of accuracy in the three stages of accuracy) Database: SwissProt

As a result, it was found that the 40 kDa proteins were Serpin Z4 derived from barley (sequence coverage: 77.2%) and Serpin Z7 derived from barley (sequence coverage: 72.8%).

(Sensory evaluation when adenosine was added to a commercially available beer-flavored alcoholic beverage 1) Adenosine was added to the commercially available beer-flavored alcoholic beverage 1, and the sensory evaluation of the swelling degree was performed. The beer-flavored alcoholic beverage is a beer-flavored alcoholic beverage in which the proportion of malt in the raw material is 50% by weight or more. The raw materials of the beer-flavored alcoholic beverage are malt and hops. As nutritional components, each 100ml contains 5.5% alcohol, 0.4-0.6g protein, 3.6g carbohydrate, and about 12.5mg purine.

The standard score of the sensory evaluation is as follows. Five professional evaluators were assigned points on a scale of 0.05 points according to the following criteria, and the score values were averaged. The swelling degree strength is based on the following criteria. 0 points: no feeling at all 1 point: Slightly felt 2 points: clearly felt 3 points: very feeling As a reference point, the beer-flavored alcoholic beverage (I) which is the same as the commercial beer-flavored alcoholic beverage 1 of the evaluation object was used as a reference, and the swelling degree was set to 1.5 points. In addition, the swelling degree of the beverage in which 25 ppm of 40 kDa protein was added and the concentration of 40 kDa protein was 50 ppm in the same beer-flavored alcoholic beverage as the above-mentioned commercially available beer-flavored alcoholic beverage 1 of the evaluation object was set to 2 points.

The order of sensory evaluation is as follows. (1) Pour beer-flavored alcoholic beverages into small glass bottles of 1/10 of the final capacity (v/v) (2) Weigh any weight of adenosine and add (3) Vibration treatment for 30 seconds (4) Stand at room temperature for 30 minutes (5) Fill the beer-flavored alcoholic beverage to the final capacity (6) Drinking evaluation after injection

(Analysis of Commercially Available Beer-Flavored Alcoholic Beverage) The concentration of adenosine contained in the commercially available beer-flavored alcoholic beverage used in the sensory evaluation was quantified by LC-MS by the following procedure. (1) Preparation of standard substance and preparation of calibration curve Regarding adenosine, each of the adenosine was diluted to the following concentrations, passed through a 0.22 μm filter, and then supplied for measurement. Final concentration: 0.001ppm, 0.025ppm, 0.050ppm, 0.100ppm, 0.200ppm, 0.300ppm, 0.500ppm, 0.750ppm, 1.000ppm (1ppm=1μg/mL) 5% (v/v) ethanol aqueous solution was used for the dilution. And, in the analysis result of the standard product, the measured value of the dilution ratio within the range of maintaining the linearity of the calibration curve (R ² >0.99) was used.

The measurement conditions of LC-MS are as follows. LC-MS: X500R manufactured by AB Sciex Separation column: HSST3 1.8μm, 2.1x150mm, manufactured by Waters Esolution: A solution: 0.1% formic acid/water, B solution: 0.1% formic acid/acetonitrile Gradient: A solution: B solution = 98: 2→2: 98 (27 minutes) 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℃

(2) Preparation of test samples from commercially available beer-flavored alcoholic beverages Commercially available beer-flavored alcoholic beverages were degassed by vibrating, and after the bubbles were relieved, they were appropriately diluted and passed through a 0.22 μm filter before being supplied to the measurement. A 5% (v/v) ethanol aqueous solution was used as the diluent. The concentration of adenosine contained in the commercially available beer-flavored alcoholic beverage 1 was taken as the control group 1 .

The concentration of the 40 kDa protein contained in the commercially available beer-flavored alcoholic beverage 1 used in the sensory evaluation was measured by a LabChip™ GXII apparatus (manufactured by PerkinElmer) using an HT Protein Express wafer according to a standard procedure (n=3). The concentration of 40 kDa protein contained in the commercially available beer-flavored alcoholic beverage 1 was 25 ppm.

(Example 1: Evaluation of Adenosine Addition) The concentration of adenosine contained in the commercially available beer-flavored alcoholic beverage 1 (control group 1) was 41.9 ppm. On the other hand, adenosine was added so that the concentration of adenosine was 50 ppm, 100 ppm, and 150 ppm, respectively, and sensory evaluation was performed (Sample 2, Sample 4, and Sample 5). The results of the sensory evaluation are shown in Table 1.

(Example 2: Evaluation of the additive effect of adenosine and 40kDa protein) 40kDa protein and adenosine were added to a commercially available beer-flavored alcoholic beverage 1 (control group 1), and 40kDa protein and adenosine were added to make the 40kDa protein concentration 30 ppm, and the adenosine concentrations were 45 ppm and 50 ppm, respectively. Evaluation test (sample 1 and sample 3). Among the 40kDa proteins, those purified above were used. In addition, for comparison, evaluation was also performed when only 5 ppm of the 40 kDa protein was added to the commercially available beer-flavored alcoholic beverage 1 and 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 concentration of each sample is shown in Table 1 below. The "purinomer concentration" in the present specification is the total amount of the four purinomer bases of adenine, guanine, xanthine, and hypoxanthine, as described above, and can be used, for example, after hydrolysis with perchloric acid using LC- MS/MS (liquid chromatography mass spectrometry) to detect.

From the results of Sample 2, Sample 4, and Sample 5 shown in Table 1, it can be seen that in a beer-flavored alcoholic beverage with a purine concentration of 5 mg/100 mL or more, if the adenosine concentration is 50 ppm or more, the average sensory evaluation Compared with the control group (1.5), the degree of swelling is effectively increased by 0.1 points or more. Thus, by setting the concentration of adenosine in a beer-flavored alcoholic beverage having a purine concentration of 5 mg/100 mL or more to a specific concentration or more, the swelling degree can be effectively enhanced relative to the control group. In addition, from the results of Comparative Sample 1, even if only 40 kDa protein was added to the beer-flavored alcoholic beverage, it was found to be 0.08 points larger than that of the control group, and the swelling degree could be enhanced. However, it can be seen that the increase value of the control group derived from the functional evaluation of sample 1 in which adenosine and 40 kDa protein are used together is 0.1 points or more. Even if the concentration of adenosine is less than 50 ppm, the swelling degree is effective by using 40 kDa protein in combination. ground is enhanced. Therefore, by using the 40 kDa protein and adenosine in combination, an unexpected synergistic effect can be exhibited in the enhancement of the degree of swelling.

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

In addition, the beer-flavored alcoholic beverage 2 of the evaluation object of Example 3 is a beer-flavored alcoholic beverage in which the ratio of the malt in the raw material is 50% by weight or more. This beer-flavored alcoholic beverage 2 is a beer-flavored alcoholic beverage different 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 each 100ml contains 5% alcohol, 0.2~0.4g protein, 3.0g sugar, and 5~6mg purine as nutrients.

The reference points and order of the sensory evaluation are the same as those in Example 1. The results of the sensory evaluation are shown in Table 2.

From the results of sample 6 shown in Table 2, it can be seen that in beer-flavored alcoholic beverage 2 with a purine concentration of 5 mg/100 mL or more, if the adenosine concentration is 50 ppm or more, even if the 40 kDa protein concentration is 6.5 ppm. For a low value, the average of the sensory evaluations is 0.07 points higher than that of the control group 2 (1.0), and the degree of swelling is enhanced. Therefore, even when the 40 kDa protein concentration is low, the degree of swelling can be enhanced relative to the control group 2 by setting the concentration of adenosine above a specific concentration. Furthermore, it can be seen that the increase value of the control group 2 from the functional evaluation of the sample 7 with the adenosine concentration of 50 ppm and the 40 kDa protein of 10 ppm is 0.1 points or more. When the concentration is above a specific concentration, the degree of swelling is effectively enhanced. Therefore, when the adenosine concentration is 50 ppm or more and the 40 kDa protein concentration is 10 ppm or more, an unexpected synergistic effect can be exhibited in the enhancement of the degree of swelling. [industrial availability]

According to the present invention, it is possible to provide a beverage with enhanced swelling in a beer-flavored alcoholic beverage having a purine concentration of 5 mg/100 mL or more.

Claims (5)

A beer-flavored alcoholic beverage, the concentration of which is a purine body of 5 mg/100 mL or more and adenosine concentration of 50 ppm or more. The beer-flavored alcoholic beverage of claim 1, wherein the concentration of adenosine is 50-150 ppm. The beer-flavored alcoholic beverage according to claim 1 or 2, further comprising a protein with a molecular weight of 35 to 50 kDa, and the concentration of the protein is 10 ppm or more. The beer-flavored alcoholic beverage according to claim 3, wherein the concentration of the protein is 200 ppm or less. A beer-flavored alcoholic beverage with a purine concentration of 5 mg/100 mL or more, comprising adenosine and a protein with a molecular weight of 35-50 kDa, wherein the adenosine concentration is 45 ppm or more, and the protein concentration is 30 ppm or more.