TW202206588A - Beer-flavored alcoholic beverage - Google Patents

Abstract

The purpose of the present invention is to provide: a beverage having an enhanced richness; a beer-flavored alcoholic beverage having an enhanced richness and a malt percentage in the raw material of at least 50 weight%; and a beer-flavored alcoholic beverage having an enhanced richness and a malt percentage in the raw material of less than 50 weight%. The present invention relates, inter alia, to a beer-flavored alcoholic beverage in which the malt percentage in the raw material is at least 50 weight% and in which the concentration of 2'-deoxyadenosine is at least 11 ppm.

Description

beer-flavored alcoholic beverages

The present invention relates to beer-flavored alcoholic beverages.

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. The object of the present invention is to provide a beer-flavored alcoholic beverage in which the proportion of malt in the raw material is 50% by weight or more and the swelling degree is enhanced. Furthermore, an object of the present invention is to provide a beer-flavored alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight and the swelling degree is enhanced. A beer-flavored alcoholic beverage having a malt ratio of less than 50% by weight tends to lack the swelling degree, and therefore, a beverage with an enhanced swelling degree is particularly desired. [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, which is a beer-flavored alcoholic beverage in which the proportion of malt in the raw material 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 the above [1], wherein the concentration of 2'-deoxyadenosine is 11-50 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, which is a beer-flavored alcoholic beverage in which the proportion of malt in the raw material is less than 50% by weight, and the concentration of 2'-deoxyadenosine is 9.2 ppm or more. [6] The beer-flavored alcoholic beverage according to the above [5], wherein the concentration of 2'-deoxyadenosine is 9.2 to 100 ppm. [7] The beer-flavored alcoholic beverage according to the above [5] or [6], further comprising a protein having a molecular weight of 35 to 50 kDa, and the concentration of the protein is 1 ppm or more. [8] The beer-flavored alcoholic beverage according to the above [7], wherein the concentration of the protein is 30 ppm or less. [Effect of invention]

According to the present invention, it is possible to provide a beer-flavored alcoholic beverage in which the ratio of malt in the raw material is 50% by weight or more and the swelling degree is enhanced. Furthermore, according to this invention, the ratio of malt in a raw material is less than 50 weight%, and the beer-flavored alcoholic beverage whose swelling degree is heightened can be provided.

The ratio of the malt in the raw material of the beer-flavored alcoholic beverage of the first aspect of the present invention is 50% by weight or more. In the beer-flavored alcoholic beverage of the first aspect of the present invention, the proportion of malt is preferably 100% by weight.

The ratio of the malt in the raw material of the beer-flavored alcoholic beverage of the second aspect of the present invention is less than 50% by weight. In the beer-flavored alcoholic beverage of the second aspect of the present invention, the proportion of malt is preferably 30% by weight or more. In addition, the ratio of malt in the raw material of the beer-flavored alcoholic beverage of the second aspect of the present invention may be 0%.

Hereinafter, in this specification, beer flavored beverages in which the proportion of malt in the raw material of the first aspect of the present invention is 50% by weight or more, and beer in which the proportion of malt in the raw material of the second aspect of the present invention is less than 50% by weight Descriptions common to flavored alcoholic beverages are simply described as "beer-flavored alcoholic beverages".

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.

The beer-flavored alcoholic beverage of the present invention contains 2'-deoxyadenosine. 2'-Deoxyadenosine is one of the deoxyribonucleosides. In this specification, 2'-deoxyadenosine may be represented as 2'DA.

In the beer-flavored alcoholic beverage in which the proportion of malt in the raw material of the first aspect of the present invention is 50% by weight or more, the concentration of 2'-deoxyadenosine is 11 ppm or more. In a beer-flavored alcoholic beverage in which the proportion of malt in the raw material is 50% by weight or more, if the concentration of 2'-deoxyadenosine is 11 ppm or more, the beer-flavored alcoholic beverage can enhance the degree of swelling. The fact that the inventors of the present invention have found that the swelling degree of a beer-flavored alcoholic beverage containing 2'-deoxyadenosine at a specific concentration or more and a ratio of malt in the raw material of 50% by weight or more has not been known until now. In the beer-flavored alcoholic beverage in which the proportion of malt in the raw material is 50% by weight or more, the concentration of 2'-deoxyadenosine is preferably 15 ppm or more.

In the beer-flavored alcoholic beverage of the first aspect of the present invention, the concentration of 2'-deoxyadenosine is preferably 50 ppm or less. This is because in a beer-flavored alcoholic beverage in which the proportion of malt in the raw material is 50% by weight or more, if the concentration of 2'-deoxyadenosine becomes too high, the bitterness of the stamina and the bitterness of the stamina may be strongly felt. / or miscellaneous flavors. In one form, the concentration of 2'-deoxyadenosine in the beer-flavored alcoholic beverage of the first form of the present invention is preferably 11-50 ppm, more preferably 15-50 ppm.

In the beer-flavored alcoholic beverage in which the proportion of malt in the raw material of the second aspect of the present invention is less than 50% by weight, the concentration of 2'-deoxyadenosine is 9.2 ppm or more. In a beer-flavored alcoholic beverage in which the proportion of malt in the raw material is less than 50% by weight, if the concentration of 2'-deoxyadenosine is 9.2 ppm or more, the beer-flavored alcoholic beverage can enhance the degree of swelling. The fact that the inventors of the present invention have discovered that the degree of swelling of beer-flavored alcoholic beverages containing 2'-deoxyadenosine at a specific concentration or more and the ratio of malt in the raw material to less than 50% by weight has not been known until now. In the beer-flavored alcoholic beverage in which the proportion of malt in the raw material is less than 50% by weight, the concentration of 2'-deoxyadenosine is preferably 9.5 ppm or more.

In the beer-flavored alcoholic beverage in which the proportion of malt in the raw material of the second aspect of the present invention is less than 50% by weight, the concentration of 2'-deoxyadenosine is preferably 100 ppm or less. This is because in a beer-flavored alcoholic beverage in which the proportion of malt in the raw material is less than 50% by weight, if the concentration of 2'-deoxyadenosine becomes too high, the bitterness of stamina may be strongly felt. In the beer-flavored alcoholic beverage in which the proportion of malt in the raw material is less than 50% by weight, the concentration of 2'-deoxyadenosine is preferably 50 ppm or less. In one form, the concentration of 2'-deoxyadenosine in the beer-flavored alcoholic beverage in which the proportion of malt in the raw material is less than 50% by weight is preferably 9.2 to 100 ppm, more preferably 9.5 to 100 ppm, and is 9.5~50ppm is better.

In the beer-flavored beverage of the present invention, it is preferable to further comprise a protein with a molecular weight of 35-50 kDa. In the beer-flavored alcoholic beverage in which the proportion of malt in the raw material of the first aspect of the present invention is 50% by weight or more, protein with a molecular weight of 35 to 50 kDa is further contained, and the concentration of the protein is preferably 10 ppm or more.

In the beer-flavored alcoholic beverage in which the proportion of malt in the raw material of the second aspect of the present invention is less than 50% by weight, a protein with a molecular weight of 35 to 50 kDa is further contained, and the concentration of the protein is preferably 1 ppm or more.

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.

By further containing 40 kDa protein in addition to 2'-deoxyadenosine, the swelling degree of the beer-flavored alcoholic beverage in which the ratio of malt in the raw material is 50% by weight or more can be enhanced. Furthermore, in the beer-flavored beverage in which the proportion of malt in the raw material of the first aspect of the present invention is 50% by weight or more, the concentration of 40kDa protein is more preferably 25ppm or more, more preferably 30ppm or more, and more preferably 200ppm or less. Preferably, it is more preferably below 100 ppm. In one form, in the beer-flavored beverage of the first form of the present invention, the concentration of 40kDa protein is preferably 10 to 200 ppm, more preferably 25 to 200 ppm, more preferably 30 to 200 ppm, and particularly preferably 30 to 100 ppm.

By further containing 40 kDa protein in addition to 2'-deoxyadenosine, the swelling degree of the beer-flavored alcoholic beverage of the second aspect of the present invention in which the proportion of malt in the raw material is less than 50% by weight can be enhanced. In the beer-flavored beverage of the second aspect of the present invention, the concentration of the 40 kDa protein is more preferably 5 ppm or more, and more preferably 30 ppm or less. In one form, in the beer-flavored beverage of the second form of the present invention, the concentration of the 40kDa protein is preferably 1-30 ppm, more preferably 5-30 ppm.

When 2'-deoxyadenosine and 40kDa protein are contained at the same time, the swelling degree of beer-flavored alcoholic beverages can be effectively enhanced by the synergistic effect of 2'-deoxyadenosine and 40kDa protein.

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 a general beer-flavored alcoholic beverage common to the first aspect and the second aspect of the present invention 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 can 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 in which the proportion of malt in the raw material of the second aspect of the present invention is less than 50% by weight may also be a beer-flavored alcoholic beverage produced by the above-mentioned method, by adding malt-derived distilled liquor (for example, distilled spirits) Manufactured from alcoholic beverages derived from wheat, such as liquor or shochu, etc.

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 aspect of the present invention, liquid sugar containing a carbon source, wheat or malt other than the amino acid-containing material are used. Nitrogen sources, hops, pigments, etc. are mixed with warm water as a 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 can be used. The obtained fermentation broth was filtered, and carbon dioxide gas was added to the obtained filtrate. After that, it was filled in a container and subjected to a sterilization step to obtain a beer-flavored alcoholic beverage in which the ratio of malt in the target raw material was less than 50% by weight.

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 measured by nitrogen quantitative conversion method, the amount of lipid was measured 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 manufacturing method of the beer-flavored alcoholic beverage of this invention is not specifically limited. For example, in a beer-flavored alcoholic beverage in which the proportion of malt in the raw material is 50% by weight or more, or in a beer-flavored alcoholic beverage in which the proportion of malt in the raw material is less than 50% by weight, a specific amount of 2'-deoxygenation is added. Adenosine method. Also, a beer-flavored alcoholic beverage in which the proportion of malt in the raw material of the first aspect of the present invention is 50% by weight or more or a beer-flavored alcoholic beverage in which the proportion of malt in the raw material of the second aspect of the present invention is less than 50% by weight It is better to add 40kDa protein.

The preparation of the added 2'-deoxyadenosine and the 40-kDa protein can be carried out, for example, by the procedure described in the examples described later. And, regarding 2'-deoxyadenosine and 40kDa protein, the content of these can also be increased by adjusting various conditions in the production process of beer-flavored alcoholic beverages. [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 2'-deoxyadenosine) Purification of 2'-deoxyadenosine (2'DA) was carried out according to the following. (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. The respective eluted fractions were refrigerated and stored as dried products using an evaporator and a freeze dryer.

(2) LH-20 Fractionation of 30% Ethanol Dissolution Fraction Among the HP20 fractions, 1.2 kg of Sephadex (registered trademark) LH-20 was used for fractionation of the 30% ethanol-eluted fraction. The LH-20 washed with ethanol was packed into a column for fractionation in large quantities, and replaced with water. 17.6 g of the 30% ethanol eluted fraction (87.9 g) obtained by fractional distillation with HP20 was dissolved in distilled water and used in an LH-20 column. Use a medium-pressure pump to flow in 13.5L of distilled water to obtain water-dissolved fractions -1~6. Next, 7 L of water-containing ethanol (35% ethanol, 70% ethanol and 100% ethanol) was poured into each to obtain elution liquids as 35% ethanol eluted fractions, 70% ethanol eluted fractions and 100% ethanol eluted fractions, respectively. The respective eluted fractions were refrigerated and stored as dried products using an evaporator and a freeze dryer.

(3) Separation of 2'DA Of the water-eluted fraction-4 (0.56 g) obtained by fractional distillation with LH-20, 86.4 mg was eluted with 10% ethanol using HPLC (COSMOSIL 5C18-PAQ, 20×250 mm). Next, the eluate was concentrated from 10 minutes to 13 minutes, and was 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 (0.5 mg, tR=21 min). Compound (I) was identified as 2'-deoxyadenosine by analysis of physical data with MS and NMR and comparison with the standard. 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) After washing with water, 10 mL of the cation exchange resin-bound fraction obtained in (1) was added to an out-of-limit filtration unit (Amicon Ultra-15 30K manufactured by Merck), and centrifuged at 3500 rpm to obtain an out-of-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. Cut out the fragment around 40kDa separated by SDS-PAGE, reduce with dithiothreitol (56°C, 1 hour), and carry out carbamoamide methylation 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% ProteaseMax, 15 μL of 5 mM calcium chloride, 50 mM ammonium bicarbonate solution was 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: A solution: 0.1% formic acid/water, B solution: 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 Measuring range: MS1 (m/z 350-1750) Data Dependent Scan mode

(4) Analysis of protein Protein identification was performed under the following conditions. Retrieval software: Proteome Discoverer 2.2.0.388 (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%).

<I. Sensory evaluation when 2'DA is added to a commercially available beer-flavored alcoholic beverage with a malt content of 50% by weight or more> In Examples 1 and 2, 2'-deoxyadenosine (2'DA) was added to a commercially available beer-flavored alcoholic beverage (A), and the sensory evaluation of swelling degree was performed. The beer-flavored alcoholic beverage (A) is a beer-flavored alcoholic beverage in which the ratio 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 standard score, the degree of swelling was set to 0.7 points for the commercially available beer-flavored alcoholic beverage (B) different from the above-mentioned commercially available beer-flavored alcoholic beverage (A) to be evaluated as the standard beer-flavored alcoholic beverage (I). In addition, the swelling degree of the beer-flavored alcoholic beverage (II) which is the same as the commercially available beer-flavored alcoholic beverage (A) of the evaluation object was set as a reference was made 1.5 points of the reference. The standard beer-flavored alcoholic beverage (I) is a beer-flavored alcoholic beverage in which the malt ratio in the raw material exceeds 0% by weight and is less than 50% by weight. The raw materials are sparkling wine, malt, hops, sugar, dietary fiber, and distilled liquor (wheat). As nutrients, each 100ml contains 4% alcohol, 0~0.2g protein, 0.5~0.8g carbohydrate, and purine. About 2.0mg.

The order of sensory evaluation is as follows. (1) Pour the beer-flavored alcoholic beverages to be evaluated into 1/10 volume (v/v) small glass bottles of the final volume. (2) Weigh 2'DA of any weight 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 2'DA 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 product and preparation of calibration curve 2'DA 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 1.8 μm, 2.1×150 mm 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 Measuring 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 2'DA contained in the commercially available beer-flavored alcoholic beverage (A) was taken as control group 1.

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

(Example 1: Evaluation of 2'DA addition) The concentration of 2'DA contained in the commercially available beer-flavored alcoholic beverage (A) (control group 1) was 8 ppm. On the other hand, 2'DA was added so that the concentration of 2'DA was 11 ppm, 15 ppm, 20 ppm, and 50 ppm, respectively, and a sensory evaluation was performed. The results of the sensory evaluation are shown in Table 1.

From the results shown in Table 1, it can be known that in the beer-flavored alcoholic beverage in which the ratio of malt in the raw material is 50% by weight or more, the swelling degree will be enhanced if the 2'DA concentration is 11 ppm or more.

(Example 2: Evaluation of the additive effect of 2'DA and 40kDa protein) 2'DA was added to a commercially available beer-flavored alcoholic beverage (A) in which the ratio of malt in the raw material was 50% by weight or more and the concentration of 2'DA was 11 ppm as a standard, and 40 kDa protein was further added to evaluate 2 'The additive effect of DA and 40kDa protein. 40kDa protein was added so that the concentrations of the 40kDa protein were 30 ppm, 35 ppm, and 45 ppm, respectively. Among the 40kDa proteins, those purified above were used. In addition, for comparison, the evaluation of adding only 5 ppm of 40 kDa protein to the commercially available beer-flavored alcoholic beverage (A) was also performed (comparative sample 2-1). The results of the sensory evaluation are shown in Table 2.

From the results shown in Table 2, it can be seen that adding 2'DA and further adding 40kDa protein to the commercially available beer-flavored alcoholic beverage (A) in which the proportion of malt in the raw material is 50% by weight or more can further enhance swelling. Spend. From the results of Comparative Sample 2-1, it can be seen that the swelling degree can be enhanced even by adding only 40 kDa protein. However, it can be seen that the sum of the increase value (0.11) of the control group from the sensory evaluation of the above-mentioned sample 2-1 and the increase value (0.06) of the control group from the sensory evaluation of the comparative sample 2-1 can be obtained, compared with The predicted additive effect (0.17) upon addition of 2'DA and 40 kDa protein was greater for the control group (0.21) from the functional evaluation of sample 2-2. By using 2'DA and 40kDa protein together, an unexpected synergistic effect can be exhibited in the enhancement of swelling degree.

(Example 3: Evaluation of the additive effect of 2'DA and 40kDa protein) A commercially available beer-flavored alcoholic beverage (referred to as a beer-flavored alcoholic beverage (C)) was added with 2'DA and 40 kDa protein, and sensory evaluation was performed. In Example 3, the beer-flavored alcoholic beverage (C) to be evaluated is a beer-flavored alcoholic beverage in which the ratio of malt in the raw material is 50% by weight or more. This beer-flavored alcoholic beverage (C) is a beer-flavored alcoholic beverage different from the commercially available beer-flavored alcoholic beverage (A) used in Examples 1 and 2. The raw materials of beer-flavored alcoholic beverage (C) are malt, hops, rice, corn, and starch. As nutrients, each 100ml contains 5% alcohol, 0.2~0.4g protein, 3.0g sugar, and 5~ purine. 6mg.

The 2'DA and 40 kDa protein concentrations contained in the beer-flavored alcoholic beverage (C) were analyzed by the above method. The concentration of 2'DA contained in the beer-flavored alcoholic beverage (C) (control group 2) was 10 ppm. Also, the concentration of 40 kDa protein was 6.5 ppm. 2'DA was added to this beer-flavored alcoholic beverage (C) so that the concentration of 2'DA was 11 ppm, and a sensory evaluation was performed (Sample 3-2). Then, 2'DA was added to the beer-flavored alcoholic beverage (C) so that the concentration of 2'DA was 11 ppm, and 40 kDa protein was further added to make the concentration of 40 kDa protein 10 ppm, and sensory evaluation was performed (Sample 3-3).

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 3.

From the results shown in Table 3, it can be seen that even if the 40kDa protein concentration is as low as 6.5ppm, the ratio of malt in the raw material is 50% by weight or more of the commercially available beer-flavored alcoholic beverage (C) by adding it. 2'DA, which can enhance swelling. Furthermore, with 2'DA of 11 ppm and 40 kDa protein concentration of 10 ppm, beer can more effectively enhance the swelling degree of flavored alcoholic beverages.

<II. Sensory evaluation when 2'DA is added to a commercially available beer-flavored alcoholic beverage (B) in which the proportion of malt in the raw material is less than 50% by weight> 2'-Deoxyadenosine (2'DA) was added to a commercially available beer-flavored alcoholic beverage (B), and the sensory evaluation of swelling degree was performed. The beer-flavored alcoholic beverage (B) is a beer-flavored alcoholic beverage in which malt is contained in the raw material, and the ratio of the malt in the raw material is less than 50% by weight. The raw materials are sparkling wine, malt, hops, sugar, dietary fiber, and distilled liquor (wheat). As nutrients, each 100ml contains 4% alcohol, 0~0.2g protein, 0.5~0.8g carbohydrate, and purine. About 2.0mg.

(Analysis of commercially available beer-flavored alcoholic beverages) The concentration of 2'DA contained in the commercially available beer-flavored alcoholic beverage (B) used in the sensory evaluation was quantified by LC-MS in the same order as described above. The concentration of 2'DA contained in the commercially available beer-flavored alcoholic beverage (B) was taken as control group 3.

(Example 4: Evaluation of 2'DA addition) The concentration of 2'DA contained in the commercially available beer-flavored alcoholic beverage (B) (control group 3) was 1.9 ppm. On the other hand, 2'DA was added so that the concentrations of 2'DA were 9.2 ppm, 9.5 ppm, 15 ppm, 20 ppm, 50 ppm, and 100 ppm, respectively, and sensory evaluation was performed. 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 4.

From the results shown in Table 4, it can be seen that in the beer-flavored alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight, if the 2'DA concentration is 9.2 ppm or more, the swelling degree will be enhanced.

(Example 5: Evaluation of the additive effect of 2'DA and 40kDa protein) The addition of 2'DA to a commercially available beer-flavored alcoholic beverage (B) (control group 3) and the concentration of 2'DA of 9.2 ppm was used as a benchmark, and the addition of 40kDa protein was further added to evaluate the addition of 2'DA and 40kDa protein. Multiply effect. The concentrations of 40 kDa protein were set at 1 ppm, 5 ppm, 10 ppm, 25 ppm. Among the 40kDa proteins, those purified above were used. Furthermore, for comparison, the evaluation of adding only 40 kDa protein to the commercially available beer-flavored alcoholic beverage (B) was also performed. The results of the sensory evaluation are shown in Table 5.

From the results shown in Table 5, it can be seen that by adding 2'DA to the commercially available beer-flavored alcoholic beverage (B) in which the ratio of malt in the raw material is less than 50% by weight, and further adding 40kDa protein, the swelling degree can be further enhanced. From the results of Comparative Samples 5-1 to 5-3, it can be seen that only adding 40kDa protein can enhance the degree of swelling. However, it can be known that, for example, the sum of the increase value (0.10) of the control group from the sensory evaluation of the above-mentioned sample 5-1 and the increase value (0.03) of the control group from the sensory evaluation of the comparative sample 5-1 can be obtained, and the corresponding The increase in the control group (0.21) from the sensory evaluation of sample 5-2 was greater than the predicted additive effect (0.13) when 2'DA was used in combination with the 40 kDa protein. The increase in sensory evaluation from the control group of Sample 5-3 (0.35) was also greater than the sum of the sensory evaluation increases from the control group of Sample 5-1 and Comparative Sample 5-2 (0.10 and 0.09, respectively) . The increase in sensory evaluation from the control group of samples 5-5 (0.47) was also greater than the sum of the sensory evaluation increases from the control group of sample 5-1 and control sample 5-3 (0.10 and 0.20, respectively) . From this result, it can be said that by using 2'DA and 40kDa protein together, the beer-flavored beverage in which the ratio of malt in the raw material is less than 50% by weight can exhibit an unexpected synergistic effect in the enhancement of the swelling degree. [industrial availability]

According to the present invention, it is possible to provide a beverage with enhanced swelling in a beer-flavored alcoholic beverage in which the proportion of malt in the raw material is 50% by weight or more. Furthermore, according to the present invention, it is possible to provide a beverage with enhanced swelling in a beer-flavored alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight.

Claims (8)

A beer-flavored alcoholic beverage, which is a beer-flavored alcoholic beverage in which the ratio of malt in the raw material is 50% by weight or more, and the concentration of 2'-deoxyadenosine is 11 ppm or more. The beer-flavored alcoholic beverage of claim 1, wherein the concentration of 2'-deoxyadenosine is 11-50 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, which is a beer-flavored alcoholic beverage in which the proportion of malt in the raw material is less than 50% by weight, and the concentration of 2'-deoxyadenosine is 9.2 ppm or more. The beer-flavored alcoholic beverage of claim 5, wherein the concentration of 2'-deoxyadenosine is 9.2-100 ppm. The beer-flavored alcoholic beverage according to claim 5 or 6, further comprising a protein with a molecular weight of 35 to 50 kDa, and the concentration of the protein is 1 ppm or more. The beer-flavored alcoholic beverage according to claim 7, wherein the concentration of the protein is 30 ppm or less.