WO2022004720A1 - Beer-taste alcoholic beverage - Google Patents

Beer-taste alcoholic beverage Download PDF

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Publication number
WO2022004720A1
WO2022004720A1 PCT/JP2021/024550 JP2021024550W WO2022004720A1 WO 2022004720 A1 WO2022004720 A1 WO 2022004720A1 JP 2021024550 W JP2021024550 W JP 2021024550W WO 2022004720 A1 WO2022004720 A1 WO 2022004720A1
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WO
WIPO (PCT)
Prior art keywords
beer
alcoholic beverage
ppm
taste alcoholic
concentration
Prior art date
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PCT/JP2021/024550
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French (fr)
Japanese (ja)
Inventor
理沙 高木
菜子 首藤
恵子 岩佐
正晃 小沢
直人 神田
嘉英 松尾
Original Assignee
サントリーホールディングス株式会社
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Priority to JP2022534041A priority Critical patent/JPWO2022004720A1/ja
Publication of WO2022004720A1 publication Critical patent/WO2022004720A1/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
    • C12C5/00—Other raw materials for the preparation of beer
    • C12C5/02—Additives for beer
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
    • C12G3/00—Preparation of other alcoholic beverages
    • C12G3/02—Preparation of other alcoholic beverages by fermentation
    • C12G3/021—Preparation of other alcoholic beverages by fermentation of botanical family Poaceae, e.g. wheat, millet, sorghum, barley, rye, or corn
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
    • C12G3/00—Preparation of other alcoholic beverages
    • C12G3/04—Preparation of other alcoholic beverages by mixing, e.g. for preparation of liqueurs

Definitions

  • the present invention relates to beer-taste alcoholic beverages.
  • beer-taste alcoholic beverages having various flavor characteristics.
  • Patent Document 1 discloses that a peptide having a specific molecular weight is contained in order to improve the flavor of a beer-taste alcoholic beverage.
  • Patent Document 2 discloses a method for increasing the drinking response of beer while reducing the purine concentration.
  • Patent Document 1 indexes such as beer-like taste, smooth taste flow, and roughness remaining in the mouth are used as evaluation indexes for beverages, and by containing a peptide of 10-20 kDa at a predetermined concentration, these indexes can be used. It is stated that a beverage with a high sensory evaluation score based on it can be obtained. Further, Patent Document 2 describes that as an evaluation index of a beverage, a comprehensive evaluation as a beer-taste beverage and a response to drinking beer is used.
  • the taste that can be felt immediately after drinking a beer-taste beverage the strength, spread, thickness, and taste of the five basic tastes, that is, the tastes that cannot be expressed by sweetness, saltiness, acidity, bitterness, and umami, are maintained.
  • the characteristic that the strength of taste is well-balanced may be preferred.
  • Such a feature is referred to as "bulge" in the present specification.
  • the beverages described in Patent Document 1 and Patent Document 2 have room for further improvement from the viewpoint of swelling, and a method for enhancing the swelling of beer-taste alcoholic beverages has been desired. In addition to swelling, beverages with enhanced other flavors have also been desired.
  • An object of the present invention is to provide a beverage having an enhanced bulge. It is also an object of the present invention to provide a beverage with enhanced swelling and beer-like bitterness. In particular, it is an object of the present invention to provide a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight and the concentration of purine is less than 5 mg / 100 mL, and the beverage has enhanced swelling. Further, in a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight and the concentration of purine is less than 5 mg / 100 mL, a beer-taste alcoholic beverage with enhanced swelling and beer-like bitterness is provided. Is also one of the objects of the present invention.
  • Beer-taste alcoholic beverages with a malt ratio of less than 50% by weight tend to have less swelling or beer-like bitterness, and purine concentrations less than 5 mg / 100 mL tend to have less swelling or beer-like bitterness. It is in. Therefore, such a beverage is a beverage for which it is particularly desired to enhance the swelling or to enhance the bitterness like beer in addition to the swelling.
  • the beer-like bitterness in the present invention is a refreshing, high-quality bitterness that is felt when drinking a beer-taste alcoholic beverage, and is a bitterness that does not remain bad in the aftertaste.
  • the present invention relates to the following beer-taste alcoholic beverages.
  • a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight, and the beverage having an enhanced swelling in the above-mentioned beverage having a purine concentration of less than 5 mg / 100 mL. can do. Further, according to the present invention, in the beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight and the concentration of purine is less than 5 mg / 100 mL, the swelling and the bitterness like beer are exhibited. Enhanced beverages can be provided.
  • the ratio of malt in the raw material is less than 50% by weight.
  • the malt ratio is preferably 30% by weight or more.
  • the ratio of malt in the raw material may be 0%.
  • the "malt ratio” refers to the ratio of the weight of malt to water and raw materials other than hops, such as malt, rice, corn, kouryan, potato, starch, wheat other than malt, and sugars.
  • ingredients that can be added in trace amounts such as acidulants, sweeteners, bitterness agents, seasonings, and flavors, are not included in the above ratio calculation.
  • the beer-taste alcoholic beverage of the present invention has a purine concentration of less than 5 mg / 100 mL.
  • purine is not particularly limited as long as it is a compound having a purine nuclear structure. Therefore, “purine” includes, for example, purine bases (adenine, guanine, xanthin, hypoxanthin), purine nucleosides (adenosine, guanosine, inosin), purine nucleotides (adenylic acid, guanylate, inosinic acid), and small molecules.
  • purine bases adenine, guanine, xanthin, hypoxanthin
  • purine nucleosides adenosine, guanosine, inosin
  • purine nucleotides adenylic acid, guanylate, inosinic acid
  • small molecules Alternatively, high molecular weight nucleic acids (oligonucleotides, polynucleotides) and
  • the "purine concentration” refers to the total amount of four purine bases of adenine, guanine, xanthine, and hypoxanthine.
  • the purine can be measured by a known method, for example, a method of detecting using LC-MS / MS (liquid chromatography-mass spectrometry) after hydrolysis with perchloric acid (“Purin in alcoholic beverages”. It can be measured by "Guide to Microanalysis", Japan Food Research Laboratories, URL: http://www.jfrl.or.jp/item/nutrition/post-31.html).
  • the concentration of purines in the beer-taste alcoholic beverage of the present invention is preferably 4.5 mg / 100 mL or less, and more preferably 4.2 mg / 100 mL or less.
  • the beer-taste alcoholic beverage of the present invention contains adenosine.
  • Adenosine is a type of nucleoside consisting of adenine and ribose.
  • the above adenosine does not contain a derivative of adenosine such as 5'deoxyadenosine.
  • the present invention is a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight, that is, a beer-taste alcoholic beverage having a purine concentration of less than 5 mg / 100 mL and an adenosine concentration of 5 to 10 ppm.
  • the present invention relates to a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight, the concentration of purine is less than 5 mg / 100 mL, and the concentration of adenosine is 12 to 18 ppm.
  • the beer-taste alcoholic beverage of the present invention having an adenosine concentration of 5 to 10 ppm is described as the first embodiment of the present invention
  • the beer-taste alcoholic beverage of the present invention having an adenosine concentration of 12 to 18 ppm is described as the first aspect of the present invention.
  • the second form will be described and described. Further, the description common to the first and second embodiments of the present invention will be described simply as "the present invention” and "the beer-taste alcoholic beverage of the present invention”.
  • the concentration of the adenosine is 5 ppm or more.
  • the concentration of adenosine is 5 ppm or more, the ratio of malt in the raw material is less than 50% by weight, and the swelling in the beer-taste alcoholic beverage in which the purine concentration is reduced to less than 5 mg / 100 mL can be enhanced. It has not been known so far that the content of adenosine in a beer-taste alcoholic beverage having a purine concentration of less than 5 mg / 100 mL is related to the swelling in the beer-taste alcoholic beverage. Is the finding found by.
  • the concentration of adenosine is 10 ppm or less. This is because if the concentration of adenosine becomes too high, the taste such as spiciness may become stronger in addition to the enhancement of swelling. Therefore, in the beer-taste alcoholic beverage of the present invention, the swelling can be enhanced without increasing the spiciness by setting the concentration of adenosine to 5 to 10 ppm. In the beer-taste alcoholic beverage according to the first aspect of the present invention, the concentration of adenosine is preferably 5 to 8 ppm.
  • the concentration of the adenosine is 12 ppm or more.
  • the concentration of adenosine is 12 ppm or more, the swelling of the beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight and the purine concentration is reduced to less than 5 mg / 100 mL is enhanced. It can also enhance the beer-like bitterness of the taste. It has not been known so far that the content of 12 ppm or more of adenosine in a beer-taste alcoholic beverage having a purine concentration of less than 5 mg / 100 mL is related to the swelling and beer-like bitterness in the beer-taste alcoholic beverage. This is the finding found by the inventors.
  • the concentration of adenosine is 18 ppm or less. This is because if the concentration of adenosine becomes too high, the bitterness of the aftertaste or the astringent taste may be strongly felt in addition to the enhancement of the bitterness like swelling and beer. Therefore, in the beer-taste alcoholic beverage of the present invention, by setting the concentration of adenosine to 12 to 18 ppm, swelling and beer-like bitterness can be enhanced without causing an increase in aftertaste bitterness or astringent taste.
  • beer-taste alcoholic beverage of the present invention further contains a protein having a molecular weight of 35 to 50 kDa, and the concentration of the protein is preferably 1 ppm or more.
  • a protein having a molecular weight of 35 to 50 kDa is a protein found in a region having a molecular weight 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, the beer-taste alcoholic beverage may be subjected to extrafiltration by using a 30 kDa cut-off membrane as a pretreatment.
  • the protein is preferably a protein having a molecular weight of 35 to 45 kDa, more preferably a protein having a molecular weight of about 40 kDa. In the present specification, a protein having a molecular weight of 35 to 50 kDa is also referred to as a 40 kDa protein.
  • the 40 kDa protein is preferably a cereal-derived protein.
  • the cereal is preferably at least one selected from the group consisting of barley, wheat, corn, rice and soybean.
  • the cereal is wheat, it can contain a known protein derived from wheat used in the production of beer-taste alcoholic beverages. Examples of such wheat include barley, wheat, rye, oats, oats, and barley, and barley is preferable. Further, either germinated wheat or ungerminated wheat may be used, but germinated malt is preferable. These may be contained alone or in combination of two or more kinds.
  • Serpin Z4 also known as BSZ4, HorvuZ4, Major endosperm albumin or Protein Z
  • Serpin Z7 derived from barley (also known as BSZ7 or HorvZ)
  • a protein having an amino acid sequence in which a part of the amino acids in the amino acid sequence is deleted, substituted, inserted and / or added may be used.
  • the swelling in beer-taste alcoholic beverages can be further enhanced.
  • the concentration of the 40 kDa protein is more preferably 5 ppm or more, and more preferably 30 ppm or less.
  • both adenosine and 40 kDa protein When both adenosine and 40 kDa protein are contained, the swelling of the beer-taste alcoholic beverage can be effectively enhanced by the synergistic effect of adenosine and 40 kDa protein. Further, when the beer-taste alcoholic beverage according to the second embodiment of the present invention contains both adenosine and 40 kDa protein, the synergistic effect of adenosine and 40 kDa protein effectively enhances the swelling and beer-like bitterness of the beer-taste alcoholic beverage. Can be made to.
  • 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 content contained in the beer-taste alcoholic beverage is not limited to fermented and non-fermented.
  • the alcohol here refers to ethanol and does not include the aliphatic alcohol described later.
  • a beer-taste beverage is a carbonated beverage with a beer-like flavor.
  • the following shows the manufacturing process of a general beer-taste alcoholic beverage common to the first and second embodiments of the present invention.
  • enzymes such as amylase are added to a mixture containing raw materials such as other grains, starch, saccharides, bitterness agents, or coloring agents and water, and glue is added. It is saccharified and saccharified, and filtered to obtain a saccharified solution. If necessary, add hops and bitterness to the saccharified solution and boil, and remove solids such as coagulated protein in a clarification tank.
  • hops may be added to a malt extract mixed with warm water and boiled.
  • Hops may be mixed at any stage from the start of boiling to the end of boiling.
  • known conditions may be used.
  • known conditions may be used.
  • the obtained fermentation broth is filtered, and carbon dioxide gas is added to the obtained filtrate. After that, it is filled in a container and subjected to a sterilization step to obtain a desired beer-taste alcoholic beverage.
  • the beer-taste alcoholic beverage is produced by adding a wheat-derived alcoholic beverage such as wheat-derived distilled liquor (for example, spirits or shochu) to the beer-taste alcoholic beverage produced by the above method. You may.
  • a beer-taste alcoholic beverage produced without using malt as a raw material is a liquid obtained by mixing liquid sugar containing a carbon source, nitrogen source as an amino acid-containing material other than wheat or malt, hops, pigments, etc. with warm water.
  • the liquid sugar solution is boiled.
  • hops may be mixed with the liquid sugar solution during boiling, not before the start of boiling.
  • hops may be added to an extract made from a raw material other than malt with warm water and boiled. Hops may be mixed at any stage from the start of boiling to the end of boiling.
  • known conditions may be used.
  • the obtained fermentation broth is filtered, and carbon dioxide gas is added to the obtained filtrate. Then, it is filled in a container and subjected to a sterilization step to obtain a beer-taste alcoholic beverage in which the ratio of malt in the target raw material is less than 50% by weight.
  • 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 alcohol for raw materials or the like.
  • the alcohol for raw materials may be added in any step from the saccharification step to the filling step.
  • the alcohol content of the beer-taste alcoholic beverage according to the present invention means the alcohol content (v / v%) in the beverage, and can be measured by any known method. It can be measured by a meter. Specifically, a sample from which carbon dioxide gas has been removed from the beverage by filtration or ultrasonic waves is prepared, and the sample is directly distilled to measure the density of the obtained distillate at 15 ° C. Converted using the "Table 2 Alcohol Content and Density (15 ° C) and Specific Weight (15/15 ° C) Conversion Table" attached to (Heisei 19 National Tax Agency Royal Instruction No. 6, revised on June 22, 2007). Can be asked. When the alcohol content is a low concentration of less than 1.0%, a commercially available alcohol measuring device or gas chromatography may be used.
  • Fatty alcohol may be added to the beer-taste alcoholic beverage according to the present invention from the viewpoint of imparting a feeling of alcohol.
  • the aliphatic alcohol is not particularly limited as long as it is known, but an aliphatic alcohol having 4 to 5 carbon atoms is preferable.
  • preferred aliphatic alcohols include 2-methyl-1-propanol and 1-butanol as those having 4 carbon atoms and 3-methyl-1-butanol and 1-pen as those having 5 carbon atoms. Examples thereof include tanol and 2-pentanol. These can be used in one type or a combination of two or more types.
  • 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.
  • the content of the aliphatic alcohol can be measured by using a headspace gas chromatograph method.
  • the sugar contained in the beer-taste alcoholic beverage according to the present invention means a sugar based on the nutrition labeling standard for foods (Ministry of Health, Labor and Welfare Notification No. 176, 2003).
  • carbohydrate refers to food obtained by removing proteins, lipids, dietary fiber, ash, alcohol and water.
  • the amount of sugar in a food is calculated by subtracting the amount of protein, lipid, dietary fiber, ash and water from the weight of the food. In this case, the amounts of protein, lipid, dietary fiber, ash and water are measured by the methods listed in the nutrition labeling standards.
  • the amount of protein is measured by the nitrogen quantitative conversion method
  • the amount of lipid is measured by the ether extraction method, the chloroform / methanol mixed solution extraction method, the Gerbel method, the acid decomposition method or the Reesegotleave method
  • the amount of dietary fiber is measured. Is measured by high-speed liquid chromatograph method or Proski method
  • the amount of ash is measured by magnesium acetate-added ashing method, direct ashing method or sulfuric acid-added ashing method
  • the amount of water is measured by Karl Fisher method, drying aid. Measure by the method, vacuum overheating drying method, normal pressure heating drying method or plastic film method.
  • the beer-taste alcoholic beverage according to the present invention may be low-carbohydrate in accordance with the recent taste for low-carbohydrate. Therefore, the sugar content of the beer-taste alcoholic beverage according to the present invention may be less than 2.5 g / 100 mL or less than 0.5 g / 100 mL. Although the lower limit is not particularly set, it is usually about 0.1 g / 100 mL, and may be, for example, 0.15 g / 100 mL or more or 0.2 g / 100 mL or more.
  • hops can be used as a part of the raw material.
  • ordinary pellet 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.
  • processed hop products such as isometric hops and reduced hops may be used. These are included in the hops used in the beer-taste alcoholic beverage according to the present invention.
  • the amount of hops added is not particularly limited, but is typically about 0.0001 to 1% by weight based on the total amount of the beverage.
  • the beer-taste alcoholic beverage according to the present invention may use other raw materials as needed, as long as the effects of the present invention are not impaired.
  • sweeteners including high-sweetness sweeteners
  • bitterness agents including high-sweetness sweeteners
  • flavors yeast extracts
  • colorants such as caramel pigments
  • plant-extracted saponin-based substances such as soybean saponin and kiraya saponin
  • plant proteins such as corn and soybean
  • Peptide-containing substances protein-based substances such as bovine serum albumin
  • seasonings such as dietary fiber and amino acids
  • antioxidants such as ascorbic acid
  • the beer-taste alcoholic beverage according to the present invention can be packed in a container.
  • the form of the container is not limited in any way, and it can be filled in a sealed container such as a bottle, a can, a barrel, or a PET bottle to make a beverage in a container.
  • the method for producing a beer-taste alcoholic beverage of the present invention is not particularly limited, but is a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight, and the concentration of purine is 5 mg / 100 mL.
  • An example is a method of adding a predetermined amount of adenosine to the above-mentioned beverage having a concentration of less than 5 mg / 100 mL so that the purine concentration does not exceed 5 mg / 100 mL.
  • the adenosine and 40 kDa protein to be added can be prepared, for example, by the procedure described in Examples described later. Further, regarding adenosine and 40 kDa protein, the content thereof may be increased by adjusting various conditions in the manufacturing process of the beer-taste alcoholic beverage.
  • Adenosine was purified according to the following.
  • Protein analysis Protein identification was performed under the following conditions.
  • Search software Proteome Discoverer 2.2.0.3888 (manufactured by Thermo Fisher) Species: Barley (Hordeum vulgare), Hops (Humulus), Yeast (Saccharomyces cerevisiae) Search condition: Digestive enzyme: Chymotrypsin Precursor ion mass error range: Monoisotopic, ⁇ 10 ppm Product ion mass error range: ⁇ 0.02 Da Maximum number of missed cribes: 5 Confidence level (Percolator): High (the most probable level of the three levels of certainty) Database: SwissProt
  • the 40 kDa protein was barley-derived Serpin Z4 (sequence coverage: 77.2%) and barley-derived Serpin Z7 (sequence coverage: 72.8%).
  • the beer-taste alcoholic beverage is a beer-taste alcoholic beverage in which malt is contained in the raw material, the ratio of malt in the raw material is less than 50% by weight, and the concentration of purine is less than 5 mg / 100 mL.
  • the raw materials are low-malt beer, malt, hops, sugars, dietary fiber, and spirits (wheat).
  • alcohol content is 4%
  • protein is 0 to 0.2 g
  • sugar is 0.5 to 0.8 g
  • purine is used. Contains about 2.0 mg.
  • the reference points for the sensory evaluation of the bulge are as follows. Five expert panels scored in increments of 0.05 points according to the following criteria, and the score values were averaged. The bulge strength is based on the following criteria. 0 points: Not felt at all 1 point: Slightly felt 2 points: Clearly felt 3 points: As a very felt reference point, the same commercially available beer-taste alcoholic beverage as the above-mentioned commercially available beer-taste alcoholic beverage to be evaluated is used. The bulge was given as 0.7 points as the standard beer-taste alcoholic beverage (I). Further, another commercially available beer-taste alcoholic beverage was used as the standard beer-taste alcoholic beverage (II), and its bulge was set to 1.5 points as the reference point.
  • the standard beer-taste alcoholic beverage (II) is a beer-taste alcoholic beverage in which the ratio of malt in the raw material is 50% by weight or more.
  • the raw materials are malt and hops, and the nutritional components include 5.5% alcohol, 0.4 to 0.6 g of protein, 3.6 g of sugar, and about 12.5 mg of purines per 100 ml.
  • the reference points for sensory evaluation of beer-like bitterness are as follows. Four specialized panels scored in increments of 0.05 points according to the following criteria, and the score values were averaged. Beer-like bitterness intensity is based on the following criteria. 0 points: Not felt at all 1 point: Slightly felt 2 points: Clearly felt 3 points: As a reference point that is very felt, the same commercially available beer-taste alcoholic beverage as the above-mentioned commercially available beer-taste alcoholic beverage to be evaluated is used. As the standard beer taste alcoholic beverage (I), the beer-like bitterness was given as 1.0 point. Further, the above-mentioned other commercially available beer-taste alcoholic beverages were used as the standard beer-taste alcoholic beverages (II), and the beer-like bitterness was set to 2.0 points as the reference point.
  • the procedure for sensory evaluation is as follows. (1) Dispense beer-taste alcoholic beverages into 1/10 volume (v / v) vials of the final volume (2) Weigh and add adenosine to any weight (3) Sonic for 30 seconds (4) Let stand at room temperature for 30 minutes (5) Fill up the beer-taste alcoholic beverage to the final volume (6) Dispense and evaluate by swallowing
  • Example 1 Evaluation of swelling by addition of adenosine and / or 40 kDa protein in the first embodiment of the present invention
  • concentration of adenosine contained in the commercially available beer-taste alcoholic beverage (control) was 4.25 ppm.
  • adenosine was added so that the adenosine concentrations were 5 ppm and 8 ppm, respectively, and sensory evaluation was performed (Sample 1 and Sample 2).
  • the concentration of 40 kDa protein contained in a commercially available beer-taste alcoholic beverage (control) was 0 ppm.
  • the "purine concentration" in the present specification is the total amount of four purine bases of adenine, guanine, xanthine, and hypoxanthine, and is, for example, LC-MS / MS (liquid chromatograph) after hydrolysis with perchloric acid. It can be detected by using a graph mass spectrometry method).
  • the adenosine (5 ppm) and 40 kDa protein obtained by the sum of the increase value (0.03) from the control of the sensory evaluation in the sample 1 and the increase value (0.05) from the control of the sensory evaluation in the comparison sample 1. Since the increase value (0.13) from the control of the sensory evaluation in sample 3 is larger than the additive effect (0.08) when (5 ppm) is used in combination, by using adenosine and 40 kDa protein in combination, It can be said that an unpredictable synergistic effect is exhibited.
  • adenosine (5 ppm) and 40 kDa protein obtained by the sum of the increase value (0.03) from the control of the sensory evaluation in the sample 1 and the increase value (0.08) from the control of the sensory evaluation in the comparison sample 2. Since the increase value (0.17) from the control of the sensory evaluation in sample 4 is larger than the additive effect (0.11) when (9 ppm) is used in combination, the combined use of adenosine and 40 kDa protein It can be said that an unexpected synergistic effect is exhibited.
  • Example 2 Evaluation of swelling due to addition of adenosine in the second embodiment of the present invention
  • concentration of adenosine contained in the commercially available beer-taste alcoholic beverage (control) was 4.25 ppm.
  • adenosine was added so that the adenosine concentrations were 8 ppm, 12 ppm, 18 ppm, and 23 ppm, respectively, and the sensory evaluation of the swelling was performed (comparison sample 4, sample 5, sample 6 and contrast sample 5). The results of the sensory evaluation are shown in Table 2.
  • the purine concentration of each sample is shown in Table 2 below.
  • the "purine concentration" in the present specification is the total amount of four purine bases of adenine, guanine, xanthine, and hypoxanthine, and is, for example, LC-MS / MS (liquid chromatograph) after hydrolysis with perchloric acid. It can be detected by using a graph mass spectrometry method).
  • the adenosine concentration is 12 ppm or more (sample) for the beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight and the purine concentration is less than 5 mg / 100 mL.
  • the bulge is increased by 0.1 points or more with respect to the evaluation of the bulge of the control, and it can be seen that the bulge is effectively enhanced.
  • the sample 6 and the contrast sample 5 in Table 2 above are compared, it is confirmed that the swelling does not effectively increase according to the increase in the concentration even if the adenosine concentration exceeds 18 ppm.
  • Example 3 Evaluation of swelling due to addition of adenosine and 40 kDa protein in the second embodiment of the present invention
  • concentration of adenosine contained in the above-mentioned commercially available beer-taste alcoholic beverage was 4.25 ppm, and the concentration of 40 kDa protein was 0 ppm.
  • adenosine was added so that the adenosine concentration was 8 ppm, 12 ppm, and 18 ppm, respectively, and 40 kDa protein was added so that the 40 kDa protein concentration was 5 ppm, and the sensory evaluation of the swelling was performed (contrast sample 8, sample 7, and). Sample 8).
  • the 40 kDa protein used was the one purified above.
  • the swelling can be further enhanced by adding adenosine and 40 kDa protein (5 ppm) to a commercially available beer-taste alcoholic beverage (Comparative Sample 8, Sample 7, and Sample 8). Further, it can be seen that the swelling can be enhanced only by adding the 40 kDa protein (contrast sample 6 and contrast sample 7). However, the adenosine (12 ppm) and 40 kDa protein obtained by the sum of the increase value (0.11) from the sensory evaluation control in the sample 5 and the increase value (0.06) from the sensory evaluation control in the contrast sample 6 are obtained.
  • the increase value (0.23) from the control of the sensory evaluation in sample 7 is larger than the additive effect (0.17) when (5 ppm) is used in combination, by using adenosine and 40 kDa protein in combination, It can be said that an unpredictable synergistic effect is exhibited.
  • the synergistic effect is determined by the sum of the increase value (0.14) from the sensory evaluation control in the sample 6 and the increase value (0.06) from the sensory evaluation control in the comparison sample 6, adenosine (18 ppm). It can also be confirmed from the fact that the increase value (0.25) from the control of the sensory evaluation in the sample 8 is larger than the additive effect (0.2) when the 40 kDa protein (5 ppm) is used in combination.
  • Example 4 Evaluation of beer-like bitterness by adding adenosine and / or 40 kDa protein in the second embodiment of the present invention
  • concentration of adenosine contained in the commercially available beer-taste alcoholic beverage (control) was 4.25 ppm
  • concentration of 40 kDa protein was 0 ppm.
  • adenosine was added so that the adenosine concentrations were 8 ppm, 12 ppm, 18 ppm, and 23 ppm, respectively, and the sensory evaluation of beer-like bitterness was performed (comparison sample 4, sample 5, sample 6 and contrast sample 5).
  • adenosine and 40 kDa protein were added to the above-mentioned beverage so that the adenosine concentration was 12 ppm, 18 ppm, and the 40 kDa protein concentration were 5 ppm, respectively, with respect to the commercially available beer-taste alcoholic beverage, and the sensory evaluation of the bitterness like beer was performed ().
  • the 40 kDa protein used was the one purified above.
  • a sensory evaluation of beer-like bitterness was performed by adding only 40 kDa protein contained in a commercially available beer-taste alcoholic beverage (contrast sample 6). The results of the sensory evaluation are shown in Table 4.
  • the purine concentration of each sample is shown in Table 4 below.
  • bitterness like beer can be further enhanced by adding adenosine and 40 kDa protein (5 ppm) to a commercially available beer-taste alcoholic beverage (Sample 7 and Sample 8). Further, it can be seen that the bitterness like beer can be enhanced only by adding 40 kDa protein (contrast sample 6). However, the adenosine (12 ppm) and 40 kDa protein obtained by the sum of the increase value (0.19) from the sensory evaluation control in the sample 5 and the increase value (0.04) from the sensory evaluation control in the comparison sample 6.
  • the increase value (0.4) from the control of the sensory evaluation in sample 7 is larger than the additive effect (0.23) when (5 ppm) is used in combination, by using adenosine and 40 kDa protein in combination, It can be said that an unpredictable synergistic effect is exhibited.
  • the synergistic effect is determined by the sum of the increase value (0.34) from the sensory evaluation control in the sample 6 and the increase value (0.04) from the sensory evaluation control in the comparison sample 6, adenosine (18 ppm). It can also be confirmed from the fact that the increase value (0.58) from the control of the sensory evaluation in the sample 8 is larger than the additive effect (0.38) when the 40 kDa protein (5 ppm) is used in combination.
  • the concentration of adenosine is 12 to 18 ppm in order to enhance the swelling and beer-like bitterness of the above beer-taste alcoholic beverage without increasing the bitterness or astringent taste of the aftertaste. Conceivable.
  • a beverage with enhanced swelling in a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight in which the ratio of malt in the raw material is less than 50% by weight and the concentration of purines is less than 5 mg / 100 mL, a beverage having enhanced swelling and beer-like bitterness is produced. Can be provided.

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Abstract

The purpose of the present invention is to provide: a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50 wt%, said beverage having a purine concentration of less than 5 mg/100 mL, wherein the beer-taste alcoholic beverage has enhanced swelling; and a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50 wt%, said beverage having a purine concentration of less than 5 mg/100 mL, wherein the beverage has enhanced swelling and beer-like flavor. The present invention relates to, inter alia, a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50 wt%, wherein the beer-taste alcoholic beverage has a purine concentration of less than 5 mg/100 mL and an adenosine concentration of 5-10 ppm.

Description

ビールテイストアルコール飲料Beer taste alcoholic beverage
本発明は、ビールテイストアルコール飲料に関する。 The present invention relates to beer-taste alcoholic beverages.
近年の消費者の嗜好の多様化にともなって、様々な香味特徴をもつビールテイストアルコール飲料の開発が望まれている。 With the diversification of consumer tastes in recent years, it is desired to develop beer-taste alcoholic beverages having various flavor characteristics.
特許文献１には、ビールテイストアルコール飲料の香味を改善するために、特定の分子量のペプチドを含有させることが開示されている。 Patent Document 1 discloses that a peptide having a specific molecular weight is contained in order to improve the flavor of a beer-taste alcoholic beverage.
また、健康志向が高まる中で、低カロリー、低糖質、低プリン体量の飲料の需要が高まっている。しかし、このような低カロリー、低糖質、低プリン体量の飲料においては、これらの成分が有する香味特徴が失われたり、原料となる麦芽が不足することによって、麦芽に由来する香味特徴が失われてしまうという問題があった。特許文献２には、プリン体濃度を低減しつつ、ビールの飲み応えを増加させる方法が開示されている。 In addition, as health consciousness increases, demand for low-calorie, low-carbohydrate, and low-purine beverages is increasing. However, in such a low-calorie, low-sugar, low-purine body beverage, the malt-derived flavor characteristics are lost due to the loss of the flavor characteristics of these components or the lack of malt as a raw material. There was a problem that it would be broken. Patent Document 2 discloses a method for increasing the drinking response of beer while reducing the purine concentration.
特開２０１６－１４９９７５号公報Japanese Unexamined Patent Publication No. 2016-149975 国際公開第２０１９／１３０４５８号International Publication No. 2019/130458
特許文献１では、飲料の評価指標として、ビールらしい味わい、スムーズな味の流れ、口内に残るざらつき、といった指標を用いており、１０－２０ｋＤａのペプチドを所定濃度含有することにより、これらの指標に基づく官能評価スコアが高い飲料が得られることが記載されている。
また、特許文献２では、飲料の評価指標として、ビールの飲み応え、ビールテイスト飲料としての総合評価を用いていることが記載されている。 In Patent Document 1, indexes such as beer-like taste, smooth taste flow, and roughness remaining in the mouth are used as evaluation indexes for beverages, and by containing a peptide of 10-20 kDa at a predetermined concentration, these indexes can be used. It is stated that a beverage with a high sensory evaluation score based on it can be obtained.
Further, Patent Document 2 describes that as an evaluation index of a beverage, a comprehensive evaluation as a beer-taste beverage and a response to drinking beer is used.
ここで、ビールテイスト飲料を飲んだ直後から感じられる味覚として、５基本味、すなわち、甘味、塩味、酸味、苦味及びうま味では表せない味覚で、味の強さ、広がり、厚み、味が持続する又は味の強さのバランスがとれているという特徴が好まれることがある。このような特徴を本明細書中では「ふくらみ」という。
特許文献１及び特許文献２に記載された飲料は、ふくらみの観点からはさらに改良の余地がある飲料であるといえ、ビールテイストアルコール飲料のふくらみを増強する方法が望まれていた。また、ふくらみに加え他の味わいが増強された飲料も望まれていた。 Here, as the taste that can be felt immediately after drinking a beer-taste beverage, the strength, spread, thickness, and taste of the five basic tastes, that is, the tastes that cannot be expressed by sweetness, saltiness, acidity, bitterness, and umami, are maintained. Alternatively, the characteristic that the strength of taste is well-balanced may be preferred. Such a feature is referred to as "bulge" in the present specification.
It can be said that the beverages described in Patent Document 1 and Patent Document 2 have room for further improvement from the viewpoint of swelling, and a method for enhancing the swelling of beer-taste alcoholic beverages has been desired. In addition to swelling, beverages with enhanced other flavors have also been desired.
本発明は、ふくらみが増強された飲料を提供することを目的とする。本発明はまた、ふくらみ及びビールらしい苦味が増強された飲料を提供することを目的とする。特に原料中の麦芽の比率が５０重量％未満であり、プリン体の濃度が５ｍｇ／１００ｍＬ未満であるビールテイストアルコール飲料において、ふくらみが増強された飲料を提供することを目的とする。また、原料中の麦芽の比率が５０重量％未満であり、プリン体の濃度が５ｍｇ／１００ｍＬ未満であるビールテイストアルコール飲料において、ふくらみ及びビールらしい苦味が増強されたビールテイストアルコール飲料を提供することも本発明の目的の一つである。麦芽の比率が５０重量％未満であるビールテイストアルコール飲料は、ふくらみやビールらしい苦味が乏しい傾向があり、さらにプリン体の濃度が５ｍｇ／１００ｍＬ未満であると、ふくらみやビールらしい苦味がより乏しい傾向にある。そのためこのような飲料はふくらみを増強すること、又は、ふくらみに加えビールらしい苦味を増強することが特に望まれる飲料である。ここで、本発明においてビールらしい苦味とは、ビールテイストアルコール飲料を飲んだときに感じられる爽快で良質な苦みであり、後味に悪く残らない苦味をいう。 An object of the present invention is to provide a beverage having an enhanced bulge. It is also an object of the present invention to provide a beverage with enhanced swelling and beer-like bitterness. In particular, it is an object of the present invention to provide a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight and the concentration of purine is less than 5 mg / 100 mL, and the beverage has enhanced swelling. Further, in a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight and the concentration of purine is less than 5 mg / 100 mL, a beer-taste alcoholic beverage with enhanced swelling and beer-like bitterness is provided. Is also one of the objects of the present invention. Beer-taste alcoholic beverages with a malt ratio of less than 50% by weight tend to have less swelling or beer-like bitterness, and purine concentrations less than 5 mg / 100 mL tend to have less swelling or beer-like bitterness. It is in. Therefore, such a beverage is a beverage for which it is particularly desired to enhance the swelling or to enhance the bitterness like beer in addition to the swelling. Here, the beer-like bitterness in the present invention is a refreshing, high-quality bitterness that is felt when drinking a beer-taste alcoholic beverage, and is a bitterness that does not remain bad in the aftertaste.
すなわち、本発明は、以下のビールテイストアルコール飲料に関する。
〔１〕原料中の麦芽の比率が５０重量％未満であるビールテイストアルコール飲料であって、プリン体の濃度が５ｍｇ／１００ｍＬ未満、アデノシンの濃度が５～１０ｐｐｍであるビールテイストアルコール飲料。
〔２〕原料中の麦芽の比率が５０重量％未満であるビールテイストアルコール飲料であって、プリン体の濃度が５ｍｇ／１００ｍＬ未満、アデノシンの濃度が１２～１８ｐｐｍであるビールテイストアルコール飲料。
〔３〕さらに、分子量３５～５０ｋＤａのタンパク質を含み、上記タンパク質の濃度が１ｐｐｍ以上である〔１〕又は〔２〕に記載のビールテイストアルコール飲料。
〔４〕上記タンパク質の濃度が３０ｐｐｍ以下である〔３〕に記載のビールテイストアルコール飲料。 That is, the present invention relates to the following beer-taste alcoholic beverages.
[1] A beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight, the concentration of purine is less than 5 mg / 100 mL, and the concentration of adenosine is 5 to 10 ppm.
[2] A beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight, the concentration of purine is less than 5 mg / 100 mL, and the concentration of adenosine is 12 to 18 ppm.
[3] The beer-taste alcoholic beverage according to [1] or [2], which further contains a protein having a molecular weight of 35 to 50 kDa and has a protein concentration of 1 ppm or more.
[4] The beer-taste alcoholic beverage according to [3], wherein the protein concentration is 30 ppm or less.
本発明によれば、原料中の麦芽の比率が５０重量％未満であるビールテイストアルコール飲料であって、プリン体の濃度が５ｍｇ／１００ｍＬ未満である上記飲料において、ふくらみが増強された飲料を提供することができる。また、本発明によれば、原料中の麦芽の比率が５０重量％未満であるビールテイストアルコール飲料であって、プリン体の濃度が５ｍｇ／１００ｍＬ未満である上記飲料において、ふくらみ及びビールらしい苦味が増強された飲料を提供することができる。 According to the present invention, there is provided a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight, and the beverage having an enhanced swelling in the above-mentioned beverage having a purine concentration of less than 5 mg / 100 mL. can do. Further, according to the present invention, in the beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight and the concentration of purine is less than 5 mg / 100 mL, the swelling and the bitterness like beer are exhibited. Enhanced beverages can be provided.
本発明のビールテイストアルコール飲料は、原料中の麦芽の比率が５０重量％未満である。
麦芽の比率は好ましくは３０重量％以上である。
また、本発明のビールテイストアルコール飲料は、原料中の麦芽の比率が０％であってもよい。
ここで「麦芽の比率」とは、麦芽、米、トウモロコシ、コウリャン、バレイショ、デンプン、麦芽以外の麦、及び糖類など、水とホップ以外の原料中に占める麦芽の重量の比率をいう。ただし、酸味料、甘味料、苦味料、調味料、香料など、微量に添加され得る成分については上記比率の計算に含めない。 In the beer-taste alcoholic beverage of the present invention, the ratio of malt in the raw material is less than 50% by weight.
The malt ratio is preferably 30% by weight or more.
Further, in the beer-taste alcoholic beverage of the present invention, the ratio of malt in the raw material may be 0%.
Here, the "malt ratio" refers to the ratio of the weight of malt to water and raw materials other than hops, such as malt, rice, corn, kouryan, potato, starch, wheat other than malt, and sugars. However, ingredients that can be added in trace amounts, such as acidulants, sweeteners, bitterness agents, seasonings, and flavors, are not included in the above ratio calculation.
本発明のビールテイストアルコール飲料は、プリン体の濃度が５ｍｇ／１００ｍＬ未満である。本明細書において、「プリン体」とは、プリン核構造を有する化合物であれば特に限定されるものではない。したがって、「プリン体」としては、たとえば、プリン塩基（アデニン、グアニン、キサンチン、ヒポキサンチン）、プリンヌクレオシド（アデノシン、グアノシン、イノシン）、プリンヌクレオチド（アデニル酸、グアニル酸、イノシン酸）、および低分子または高分子の核酸（オリゴヌクレオチド、ポリヌクレオチド）などが挙げられる。
なお、本明細書中、「プリン体濃度」とは、アデニン、グアニン、キサンチン、ヒポキサンチンのプリン体塩基４種の総量を指す。プリン体の測定は公知の方法によって行うことができ、例えば、過塩素酸による加水分解後にＬＣ－ＭＳ／ＭＳ（液体クロマトグラフィー質量分析法）を用いて検出する方法（「酒類中のプリン体の微量分析のご案内」、一般財団法人・日本食品分析センター、ＵＲＬ：ｈｔｔｐ：／／ｗｗｗ．ｊｆｒｌ．ｏｒ．ｊｐ／ｉｔｅｍ／ｎｕｔｒｉｔｉｏｎ／ｐｏｓｔ－３１．ｈｔｍｌ　参照）により測定することができる。 The beer-taste alcoholic beverage of the present invention has a purine concentration of less than 5 mg / 100 mL. As used herein, the term "purine" is not particularly limited as long as it is a compound having a purine nuclear structure. Therefore, "purine" includes, for example, purine bases (adenine, guanine, xanthin, hypoxanthin), purine nucleosides (adenosine, guanosine, inosin), purine nucleotides (adenylic acid, guanylate, inosinic acid), and small molecules. Alternatively, high molecular weight nucleic acids (oligonucleotides, polynucleotides) and the like can be mentioned.
In the present specification, the "purine concentration" refers to the total amount of four purine bases of adenine, guanine, xanthine, and hypoxanthine. The purine can be measured by a known method, for example, a method of detecting using LC-MS / MS (liquid chromatography-mass spectrometry) after hydrolysis with perchloric acid (“Purin in alcoholic beverages”. It can be measured by "Guide to Microanalysis", Japan Food Research Laboratories, URL: http://www.jfrl.or.jp/item/nutrition/post-31.html).
本発明のビールテイストアルコール飲料におけるプリン体の濃度は、４．５ｍｇ／１００ｍＬ以下が好ましく、４．２ｍｇ／１００ｍＬ以下がさらに好ましい。 The concentration of purines in the beer-taste alcoholic beverage of the present invention is preferably 4.5 mg / 100 mL or less, and more preferably 4.2 mg / 100 mL or less.
本発明のビールテイストアルコール飲料は、アデノシンを含む。アデノシンは、アデニンとリボースとからなるヌクレオシドの一種である。
なお、上記アデノシンは、５’デオキシアデノシン等のアデノシンの誘導体を含むものではない。本発明は、原料中の麦芽の比率が５０重量％未満であるビールテイストアルコール飲料であって、プリン体の濃度が５ｍｇ／１００ｍＬ未満、アデノシンの濃度が５～１０ｐｐｍであるビールテイストアルコール飲料、又は、原料中の麦芽の比率が５０重量％未満であるビールテイストアルコール飲料であって、プリン体の濃度が５ｍｇ／１００ｍＬ未満、アデノシンの濃度が１２～１８ｐｐｍであるビールテイストアルコール飲料に関する。本明細書において、アデノシン濃度が５～１０ｐｐｍである本発明のビールテイストアルコール飲料を本発明の第一形態と記載し、アデノシン濃度が１２～１８ｐｐｍである本発明のビールテイストアルコール飲料を本発明の第二形態と記載して説明する。また、本発明の第一形態及び第二形態に共通する説明は、単に、「本発明」や「本発明のビールテイストアルコール飲料」と記載して説明する。 The beer-taste alcoholic beverage of the present invention contains adenosine. Adenosine is a type of nucleoside consisting of adenine and ribose.
The above adenosine does not contain a derivative of adenosine such as 5'deoxyadenosine. The present invention is a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight, that is, a beer-taste alcoholic beverage having a purine concentration of less than 5 mg / 100 mL and an adenosine concentration of 5 to 10 ppm. The present invention relates to a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight, the concentration of purine is less than 5 mg / 100 mL, and the concentration of adenosine is 12 to 18 ppm. In the present specification, the beer-taste alcoholic beverage of the present invention having an adenosine concentration of 5 to 10 ppm is described as the first embodiment of the present invention, and the beer-taste alcoholic beverage of the present invention having an adenosine concentration of 12 to 18 ppm is described as the first aspect of the present invention. The second form will be described and described. Further, the description common to the first and second embodiments of the present invention will be described simply as "the present invention" and "the beer-taste alcoholic beverage of the present invention".
本発明の第一形態におけるビールテイストアルコール飲料では、上記アデノシンの濃度が５ｐｐｍ以上である。
アデノシンの濃度が５ｐｐｍ以上であると、原料中の麦芽の比率が５０重量％未満であり、プリン体濃度が５ｍｇ／１００ｍＬ未満に低減されているビールテイストアルコール飲料におけるふくらみを増強することができる。プリン体の濃度が５ｍｇ／１００ｍL未満であるビールテイストアルコール飲料において、アデノシンを所定濃度以上含有することとビールテイストアルコール飲料におけるふくらみが関連することは、これまで知られておらず、本発明者らが見出した知見である。 In the beer-taste alcoholic beverage according to the first aspect of the present invention, the concentration of the adenosine is 5 ppm or more.
When the concentration of adenosine is 5 ppm or more, the ratio of malt in the raw material is less than 50% by weight, and the swelling in the beer-taste alcoholic beverage in which the purine concentration is reduced to less than 5 mg / 100 mL can be enhanced. It has not been known so far that the content of adenosine in a beer-taste alcoholic beverage having a purine concentration of less than 5 mg / 100 mL is related to the swelling in the beer-taste alcoholic beverage. Is the finding found by.
本発明の第一形態におけるビールテイストアルコール飲料では、アデノシンの濃度が１０ｐｐｍ以下である。
アデノシンの濃度が高くなりすぎると、ふくらみの増強の他に、辛みといった味わいが強くなる場合があるためである。よって、本発明のビールテイストアルコール飲料は、アデノシンの濃度を５～１０ｐｐｍとすることにより、辛みの増加を招くことなく、ふくらみを増強することができる。
本発明の第一形態におけるビールテイストアルコール飲料では、アデノシンの濃度は、好ましくは５～８ｐｐｍである。 In the beer-taste alcoholic beverage according to the first aspect of the present invention, the concentration of adenosine is 10 ppm or less.
This is because if the concentration of adenosine becomes too high, the taste such as spiciness may become stronger in addition to the enhancement of swelling. Therefore, in the beer-taste alcoholic beverage of the present invention, the swelling can be enhanced without increasing the spiciness by setting the concentration of adenosine to 5 to 10 ppm.
In the beer-taste alcoholic beverage according to the first aspect of the present invention, the concentration of adenosine is preferably 5 to 8 ppm.
本発明の第二形態におけるビールテイストアルコール飲料では、上記アデノシンの濃度が１２ｐｐｍ以上である。
アデノシンの濃度が１２ｐｐｍ以上であると、原料中の麦芽の比率が５０重量％未満であるビールテイストアルコール飲料であって、プリン体濃度が５ｍｇ／１００ｍＬ未満に低減されている上記飲料におけるふくらみを増強することができ、また、味わいにおけるビールらしい苦味も増強することができる。プリン体の濃度が５ｍｇ／１００ｍＬ未満であるビールテイストアルコール飲料において、アデノシンを１２ｐｐｍ以上含有することとビールテイストアルコール飲料におけるふくらみ及びビールらしい苦味が関連することは、これまで知られておらず、本発明者らが見出した知見である。 In the beer-taste alcoholic beverage according to the second embodiment of the present invention, the concentration of the adenosine is 12 ppm or more.
When the concentration of adenosine is 12 ppm or more, the swelling of the beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight and the purine concentration is reduced to less than 5 mg / 100 mL is enhanced. It can also enhance the beer-like bitterness of the taste. It has not been known so far that the content of 12 ppm or more of adenosine in a beer-taste alcoholic beverage having a purine concentration of less than 5 mg / 100 mL is related to the swelling and beer-like bitterness in the beer-taste alcoholic beverage. This is the finding found by the inventors.
本発明の第二形態におけるビールテイストアルコール飲料では、アデノシンの濃度が１８ｐｐｍ以下である。
アデノシンの濃度が高くなりすぎると、ふくらみやビールらしい苦味の増強の他に、後味の苦み又は収斂味が強く感じられる場合があるためである。よって、本発明のビールテイストアルコール飲料は、アデノシンの濃度を１２～１８ｐｐｍとすることにより、後味の苦味又は収斂味の増加を招くことなく、ふくらみ及びビールらしい苦味を増強することができる。 In the beer-taste alcoholic beverage according to the second embodiment of the present invention, the concentration of adenosine is 18 ppm or less.
This is because if the concentration of adenosine becomes too high, the bitterness of the aftertaste or the astringent taste may be strongly felt in addition to the enhancement of the bitterness like swelling and beer. Therefore, in the beer-taste alcoholic beverage of the present invention, by setting the concentration of adenosine to 12 to 18 ppm, swelling and beer-like bitterness can be enhanced without causing an increase in aftertaste bitterness or astringent taste.
また、本発明のビールテイストアルコール飲料では、さらに、分子量３５～５０ｋＤａのタンパク質を含み、上記タンパク質の濃度が１ｐｐｍ以上であることが好ましい。 Further, the beer-taste alcoholic beverage of the present invention further contains a protein having a molecular weight of 35 to 50 kDa, and the concentration of the protein is preferably 1 ppm or more.
分子量３５～５０ｋＤａのタンパク質は、ビールテイストアルコール飲料をＳＤＳ－ＰＡＧＥによる電気泳動に供した場合に分子量が３５～５０ｋＤａの領域にみられるタンパク質である。ビールテイストアルコール飲料をＳＤＳ－ＰＡＧＥによる電気泳動に供する前に、例えば、前処理としてビールテイストアルコール飲料に対して３０ｋＤａのカットオフ膜を用いて限外濾過を行ってもよい。
上記タンパク質は、好ましくは分子量が３５～４５ｋＤａのタンパク質であり、より好ましくは約４０ｋＤａのタンパク質である。本明細書では分子量３５～５０ｋＤａのタンパク質を４０ｋＤａタンパク質ともいう。 A protein having a molecular weight of 35 to 50 kDa is a protein found in a region having a molecular weight 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, the beer-taste alcoholic beverage may be subjected to extrafiltration by using a 30 kDa cut-off membrane as a pretreatment.
The protein is preferably a protein having a molecular weight of 35 to 45 kDa, more preferably a protein having a molecular weight of about 40 kDa. In the present specification, a protein having a molecular weight of 35 to 50 kDa is also referred to as a 40 kDa protein.
４０ｋＤａタンパク質は、穀類由来タンパク質であることが好ましい。
上記穀類は、大麦、小麦、トウモロコシ、イネ、大豆からなる群より選択される少なくとも１種であることが好ましい。
また、穀類が麦である場合、ビールテイストアルコール飲料の製造に使用される公知の麦に由来するタンパク質を含有することができる。このような麦としては、大麦、小麦、ライ麦、カラス麦、オート麦、エン麦などが挙げられ、好ましくは大麦である。また、発芽した麦、未発芽の麦のいずれでもよいが、好ましくは発芽した麦の麦芽である。これらは、単独で含有していてもよく、２種以上を組み合わせて含有していてもよい。 The 40 kDa protein is preferably a cereal-derived protein.
The cereal is preferably at least one selected from the group consisting of barley, wheat, corn, rice and soybean.
When the cereal is wheat, it can contain a known protein derived from wheat used in the production of beer-taste alcoholic beverages. Examples of such wheat include barley, wheat, rye, oats, oats, and barley, and barley is preferable. Further, either germinated wheat or ungerminated wheat may be used, but germinated malt is preferable. These may be contained alone or in combination of two or more kinds.
また、４０ｋＤａタンパク質として、大麦（学名：Ｈｏｒｄｅｕｍ　ｖｕｌｇａｒｅ）由来のＳｅｒｐｉｎ　Ｚ４（別名：ＢＳＺ４、ＨｏｒｖｕＺ４、Ｍａｊｏｒ　ｅｎｄｏｓｐｅｒｍ　ａｌｂｕｍｉｎ又はＰｒｏｔｅｉｎ　Ｚ）、大麦由来Ｓｅｒｐｉｎ　Ｚ７（別名：ＢＳＺ７又はＨｏｒｖｕＺ７）が好ましい。また、上記タンパク質において、そのアミノ酸配列の一部のアミノ酸が欠失、置換、挿入及び／又は付加されたアミノ酸配列を有するタンパク質であってもよい。 Further, as the 40 kDa protein, Serpin Z4 (also known as BSZ4, HorvuZ4, Major endosperm albumin or Protein Z) derived from barley (scientific name: Hordeum bulgare), Serpin Z7 derived from barley (also known as BSZ7 or HorvZ) are preferable. Further, in the above protein, a protein having an amino acid sequence in which a part of the amino acids in the amino acid sequence is deleted, substituted, inserted and / or added may be used.
アデノシンに加えて４０ｋＤａタンパク質をさらに含有することによって、ビールテイストアルコール飲料におけるふくらみをさらに増強させることができる。
また、４０ｋＤａタンパク質の濃度は５ｐｐｍ以上であることがより好ましく、また、３０ｐｐｍ以下であることが好ましい。 By further containing 40 kDa protein in addition to adenosine, the swelling in beer-taste alcoholic beverages can be further enhanced.
Further, the concentration of the 40 kDa protein is more preferably 5 ppm or more, and more preferably 30 ppm or less.
また、アデノシンと４０ｋＤａタンパク質をともに含有する場合には、アデノシンと４０ｋＤａタンパク質の相乗効果によりビールテイストアルコール飲料のふくらみを効果的に増強させることができる。また、本発明の第二形態におけるビールテイストアルコール飲料において、アデノシンと４０ｋＤａタンパク質をともに含有する場合には、アデノシンと４０ｋＤａタンパク質の相乗効果によりビールテイストアルコール飲料のふくらみ及びビールらしい苦味を効果的に増強させることができる。 When both adenosine and 40 kDa protein are contained, the swelling of the beer-taste alcoholic beverage can be effectively enhanced by the synergistic effect of adenosine and 40 kDa protein. Further, when the beer-taste alcoholic beverage according to the second embodiment of the present invention contains both adenosine and 40 kDa protein, the synergistic effect of adenosine and 40 kDa protein effectively enhances the swelling and beer-like bitterness of the beer-taste alcoholic beverage. Can be made to.
本発明のビールテイストアルコール飲料は、アルコールを含有するビールテイスト飲料であり、アルコール濃度は１％（ｖ／ｖ）～１０％（ｖ／ｖ）が好ましいが、特に限定されるものではない。さらに、該ビールテイストアルコール飲料に含まれるアルコール分の由来としては、醗酵、非醗酵に限定されるものではない。なお、ここでのアルコールはエタノールを指し、後記の脂肪族アルコールは含まれない。
ビールテイスト飲料とは、ビール様の風味をもつ炭酸飲料をいう。 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 content contained in the beer-taste alcoholic beverage is not limited to fermented and non-fermented. The alcohol here refers to ethanol and does not include the aliphatic alcohol described later.
A beer-taste beverage is a carbonated beverage with a beer-like flavor.
以下に、本発明の第一形態及び第二形態に共通する一般的なビールテイストアルコール飲料の製造工程を示す。
まず、麦芽等の麦の他、必要に応じて他の穀物、でんぷん、糖類、苦味料、又は着色料などの原料及び水を含む混合物に、必要に応じてアミラーゼなどの酵素を添加し、糊化、糖化を行なわせ、ろ過し、糖化液とする。必要に応じてホップや苦味料などを糖化液に加えて煮沸し、清澄タンクにて凝固タンパク質などの固形分を取り除く。この糖化液の代替として、麦芽エキスに温水を加えたものにホップを加えて煮沸してもよい。ホップは煮沸開始から煮沸終了前のどの段階で混合してもよい。糖化工程、煮沸工程、固形分除去工程などにおける条件は、知られている条件を用いればよい。醗酵・貯酒工程などにおける条件は、知られている条件を用いればよい。得られた醗酵液を濾過し、得られた濾過液に炭酸ガスを加える。その後、容器に充填し殺菌工程を経て目的のビールテイストアルコール飲料を得る。
ビールテイストアルコール飲料は、上記のような方法で製造されたビールテイストアルコール飲料に麦由来の蒸留酒（例えば、スピリッツや焼酎など）等の麦由来のアルコール飲料を添加して製造されたものであってもよい。 The following shows the manufacturing process of a general beer-taste alcoholic beverage common to the first and second embodiments of the present invention.
First, in addition to wheat such as malt, if necessary, enzymes such as amylase are added to a mixture containing raw materials such as other grains, starch, saccharides, bitterness agents, or coloring agents and water, and glue is added. It is saccharified and saccharified, and filtered to obtain a saccharified solution. If necessary, add hops and bitterness to the saccharified solution and boil, and remove solids such as coagulated protein in a clarification tank. As an alternative to this saccharified solution, hops may be added to a malt extract mixed with warm water and boiled. Hops may be mixed at any stage from the start of boiling to the end of boiling. As the conditions in the saccharification step, the boiling step, the solid content removing step and the like, known conditions may be used. As the conditions in the fermentation / liquor storage process and the like, known conditions may be used. The obtained fermentation broth is filtered, and carbon dioxide gas is added to the obtained filtrate. After that, it is filled in a container and subjected to a sterilization step to obtain a desired beer-taste alcoholic beverage.
The beer-taste alcoholic beverage is produced by adding a wheat-derived alcoholic beverage such as wheat-derived distilled liquor (for example, spirits or shochu) to the beer-taste alcoholic beverage produced by the above method. You may.
麦芽を原料として使用せずに製造されるビールテイストアルコール飲料は、炭素源を含有する液糖、麦又は麦芽以外のアミノ酸含有材料としての窒素源、ホップ、色素等を、温水と共に混合し、液糖溶液とする。該液糖溶液は、煮沸する。原料としてホップを用いる場合、ホップは煮沸開始前ではなく、煮沸中に、該液糖溶液に混合してもよい。この糖化液の代替として、麦芽以外の原料を用いたエキスに温水を加えたものにホップを加えて煮沸してもよい。ホップは煮沸開始から煮沸終了前のどの段階で混合してもよい。醗酵・貯酒工程などにおける条件は、知られている条件を用いればよい。得られた醗酵液を濾過し、得られた濾過液に炭酸ガスを加える。その後、容器に充填し殺菌工程を経て目的の原料中の麦芽の比率が５０重量％未満であるビールテイストアルコール飲料を得る。 A beer-taste alcoholic beverage produced without using malt as a raw material is a liquid obtained by mixing liquid sugar containing a carbon source, nitrogen source as an amino acid-containing material other than wheat or malt, hops, pigments, etc. with warm water. Use a sugar solution. The liquid sugar solution is boiled. When hops are used as a raw material, the hops may be mixed with the liquid sugar solution during boiling, not before the start of boiling. As an alternative to this saccharified solution, hops may be added to an extract made from a raw material other than malt with warm water and boiled. Hops may be mixed at any stage from the start of boiling to the end of boiling. As the conditions in the fermentation / liquor storage process and the like, known conditions may be used. The obtained fermentation broth is filtered, and carbon dioxide gas is added to the obtained filtrate. Then, it is filled in a container and subjected to a sterilization step to obtain a beer-taste alcoholic beverage in which the ratio of malt in the target raw material is less than 50% by weight.
非醗酵かつアルコールを含有するビールテイストアルコール飲料は、原料用アルコールなどを加えることにより最終製品のアルコール分を調整したものでもよい。原料用アルコールの添加は、糖化工程から充填工程までのどの工程で行ってもよい。 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 alcohol for raw materials or the like. The alcohol for raw materials may be added in any step from the saccharification step to the filling step.
本発明に係るビールテイストアルコール飲料のアルコール度数は、飲料中のアルコール分の含有量（ｖ／ｖ％）を意味し、公知のいずれの方法によっても測定することができるが、例えば、振動式密度計によって測定することができる。具体的には、飲料から濾過又は超音波によって炭酸ガスを抜いた試料を調製し、そして、その試料を直火蒸留し、得られた留液の１５℃における密度を測定し、国税庁所定分析法（平１９国税庁訓令第６号、平成１９年６月２２日改訂）の付表である「第２表アルコール分と密度（１５℃）及び比重（１５／１５℃）換算表」を用いて換算して求めることができる。アルコール度が１．０％未満の低濃度の場合は、市販のアルコール測定装置や、ガスクロマトグラフィーを用いても良い。 The alcohol content of the beer-taste alcoholic beverage according to the present invention means the alcohol content (v / v%) in the beverage, and can be measured by any known method. It can be measured by a meter. Specifically, a sample from which carbon dioxide gas has been removed from the beverage by filtration or ultrasonic waves is prepared, and the sample is directly distilled to measure the density of the obtained distillate at 15 ° C. Converted using the "Table 2 Alcohol Content and Density (15 ° C) and Specific Weight (15/15 ° C) Conversion Table" attached to (Heisei 19 National Tax Agency Royal Instruction No. 6, revised on June 22, 2007). Can be asked. When the alcohol content is a low concentration of less than 1.0%, a commercially available alcohol measuring device or gas chromatography may be used.
本発明に係るビールテイストアルコール飲料に、酒感を付与する観点から、脂肪族アルコールを添加してもよい。脂肪族アルコールとしては、公知のものであれば特に制限されないが、炭素数４～５の脂肪族アルコールが好ましい。本発明において、好ましい脂肪族アルコールとしては、炭素数４のものとして、２－メチル－１－プロパノール、１－ブタノール等が、炭素数５のものとして、３－メチル－１－ブタノール、１－ペンタノール、２－ペンタノール等が挙げられる。これらは１種又は２種以上の組み合せで用いることができる。
炭素数４～５の脂肪族アルコールの含有量は好ましくは０．０００２～０．０００７重量％であり、より好ましくは０．０００３～０．０００６重量％である。本明細書において、脂肪族アルコールの含有量は、ヘッドスペースガスクロマトグラフ法を用いて測定することができる。 Fatty alcohol may be added to the beer-taste alcoholic beverage according to the present invention from the viewpoint of imparting a feeling of alcohol. The aliphatic alcohol is not particularly limited as long as it is known, but an aliphatic alcohol having 4 to 5 carbon atoms is preferable. In the present invention, preferred aliphatic alcohols include 2-methyl-1-propanol and 1-butanol as those having 4 carbon atoms and 3-methyl-1-butanol and 1-pen as those having 5 carbon atoms. Examples thereof include tanol and 2-pentanol. These can be used in one type or a combination of two or more types.
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 by using a headspace gas chromatograph method.
本発明に係るビールテイストアルコール飲料に含まれる糖質とは、食品の栄養表示基準（平成１５年厚生労働省告示第１７６号）に基づく糖質をいう。具体的には、糖質は、食品から、タンパク質、脂質、食物繊維、灰分、アルコール分及び水分を除いたものをいう。また、食品中の糖質の量は、当該食品の重量から、タンパク質、脂質、食物繊維、灰分及び水分の量を控除することにより算定される。この場合に、タンパク質、脂質、食物繊維、灰分及び水分の量は、栄養表示基準に掲げる方法により測定する。具体的には、タンパク質の量は窒素定量換算法で測定し、脂質の量はエーテル抽出法、クロロホルム・メタノール混液抽出法、ゲルベル法、酸分解法またはレーゼゴットリーブ法で測定し、食物繊維の量は高速液体クロマトグラフ法またはプロスキー法で測定し、灰分の量は酢酸マグネシウム添加灰化法、直接灰化法または硫酸添加灰化法で測定し、水分の量はカールフィッシャー法、乾燥助剤法、減圧過熱乾燥法、常圧加熱乾燥法またはプラスチックフィルム法で測定する。 The sugar contained in the beer-taste alcoholic beverage according to the present invention means a sugar based on the nutrition labeling standard for foods (Ministry of Health, Labor and Welfare Notification No. 176, 2003). Specifically, carbohydrate refers to food obtained by removing proteins, lipids, dietary fiber, ash, alcohol and water. The amount of sugar in a food is calculated by subtracting the amount of protein, lipid, dietary fiber, ash and water from the weight of the food. In this case, the amounts of protein, lipid, dietary fiber, ash and water are measured by the methods listed in the nutrition labeling standards. Specifically, the amount of protein is measured by the nitrogen quantitative conversion method, the amount of lipid is measured by the ether extraction method, the chloroform / methanol mixed solution extraction method, the Gerbel method, the acid decomposition method or the Reesegotleave method, and the amount of dietary fiber is measured. Is measured by high-speed liquid chromatograph method or Proski method, the amount of ash is measured by magnesium acetate-added ashing method, direct ashing method or sulfuric acid-added ashing method, and the amount of water is measured by Karl Fisher method, drying aid. Measure by the method, vacuum overheating drying method, normal pressure heating drying method or plastic film method.
本発明に係るビールテイストアルコール飲料は、近年の低糖質嗜好に合わせて、低糖質であってもよい。従って、本発明に係るビールテイストアルコール飲料の糖質の含有量は、２．５ｇ／１００ｍＬ未満であってもよく、０．５ｇ／１００ｍＬ未満であってもよい。また、下限は特に設定されないが、通常、０．１ｇ／１００ｍＬ程度であり、例えば、０．１５ｇ／１００ｍＬ以上であっても、０．２ｇ／１００ｍＬ以上であってもよい。 The beer-taste alcoholic beverage according to the present invention may be low-carbohydrate in accordance with the recent taste for low-carbohydrate. Therefore, the sugar content of the beer-taste alcoholic beverage according to the present invention may be less than 2.5 g / 100 mL or less than 0.5 g / 100 mL. Although the lower limit is not particularly set, it is usually about 0.1 g / 100 mL, and may be, for example, 0.15 g / 100 mL or more or 0.2 g / 100 mL or more.
本発明に係るビールテイストアルコール飲料においては、原料の一部にホップを用いることができる。
ホップを使用する際には、ビール等の製造に使用される通常のペレットホップ、粉末ホップ、ホップエキスを、所望の香味に応じて適宜選択して使用することができる。また、イソ化ホップ、還元ホップなどのホップ加工品を用いてもよい。本発明に係るビールテイストアルコール飲料に使用されるホップには、これらのものが包含される。また、ホップの添加量は特に限定されないが、典型的には、飲料全量に対して０．０００１～１重量％程度である。 In the beer-taste alcoholic beverage according to the present invention, hops can be used as a part of the raw material.
When hops are used, ordinary pellet 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. Further, processed hop products such as isometric hops and reduced hops may be used. These are included in the hops used in the beer-taste alcoholic beverage according to the present invention. The amount of hops added is not particularly limited, but is typically about 0.0001 to 1% by weight based on the total amount of the beverage.
本発明に係るビールテイストアルコール飲料は、本発明の効果を妨げない範囲で、必要に応じて、その他の原料を用いてもよい。例えば、甘味料（高甘味度甘味料を含む）、苦味料、香料、酵母エキス、カラメル色素などの着色料、大豆サポニンやキラヤサポニン等の植物抽出サポニン系物質、コーンや大豆などの植物タンパク質およびペプチド含有物、ウシ血清アルブミン等のタンパク質系物質、食物繊維やアミノ酸などの調味料、アスコルビン酸等の酸化防止剤を、本発明の効果を妨げない範囲で必要に応じて用いることができる。 The beer-taste alcoholic beverage according to the present invention may use other raw materials as needed, as long as the effects of the present invention are not impaired. For example, sweeteners (including high-sweetness sweeteners), bitterness agents, flavors, yeast extracts, colorants such as caramel pigments, plant-extracted saponin-based substances such as soybean saponin and kiraya saponin, plant proteins such as corn and soybean, and Peptide-containing substances, protein-based substances such as bovine serum albumin, seasonings such as dietary fiber and amino acids, and antioxidants such as ascorbic acid can be used as needed as long as they do not interfere with the effects of the present invention.
本発明に係るビールテイストアルコール飲料は、容器詰めとすることができる。容器の形態は何ら制限されず、ビン、缶、樽、またはペットボトル等の密封容器に充填して、容器入り飲料とすることができる。 The beer-taste alcoholic beverage according to the present invention can be packed in a container. The form of the container is not limited in any way, and it can be filled in a sealed container such as a bottle, a can, a barrel, or a PET bottle to make a beverage in a container.
本発明のビールテイストアルコール飲料の製造方法は、特に限定されるものではないが、原料中の麦芽の比率が５０重量％未満であるビールテイストアルコール飲料であって、プリン体の濃度が５ｍｇ／１００ｍＬ未満である上記飲料に対して、プリン体濃度が５ｍｇ／１００ｍＬを超えないようにアデノシンを所定量添加する方法が例示される。
また、原料中の麦芽の比率が５０重量％未満であるビールテイストアルコール飲料であって、プリン体の濃度が５ｍｇ／１００ｍＬ未満である上記飲料に対して、４０ｋＤａタンパク質を添加することが好ましい。
添加するアデノシン及び４０ｋＤａタンパク質の調製は、例えば、後述する実施例に記載の手順で行うことができる。
また、アデノシン及び４０ｋＤａタンパク質については、ビールテイストアルコール飲料の製造過程における諸条件を調整することによって、これらの含有量が増えるようにしてもよい。 The method for producing a beer-taste alcoholic beverage of the present invention is not particularly limited, but is a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight, and the concentration of purine is 5 mg / 100 mL. An example is a method of adding a predetermined amount of adenosine to the above-mentioned beverage having a concentration of less than 5 mg / 100 mL so that the purine concentration does not exceed 5 mg / 100 mL.
Further, it is preferable to add 40 kDa protein to the beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight and the concentration of purine is less than 5 mg / 100 mL.
The adenosine and 40 kDa protein to be added can be prepared, for example, by the procedure described in Examples described later.
Further, regarding adenosine and 40 kDa protein, the content thereof may be increased by adjusting various conditions in the manufacturing process of the beer-taste alcoholic beverage.
以下、実施例を示して本発明を具体的に説明するが、本発明は下記実施例に制限されるものではない。 Hereinafter, the present invention will be specifically described with reference to Examples, but the present invention is not limited to the following Examples.
（アデノシンの精製）
下記に従いアデノシンの精製を行った。
（１）ビールのＨＰ２０による分画
６０Ｌのビールを、１０Ｌのダイヤイオン（登録商標）ＨＰ２０（三菱ケミカル株式会社製）を用いて分画した。ＨＰ２０は使用前に、エタノールにより３回洗浄し、次いで５０％エタノールにより３回洗浄した。洗浄したＨＰ２０を大量分画用カラムへ充填し、水により置換した。脱気させた６０Ｌのビールへ同量の蒸留水を混合し、中圧ポンプを用いてＨＰ２０カラムへ流した。ＨＰ２０カラムを素通りした溶液を素通り画分として得た。中圧ポンプを用いて４０Ｌの蒸留水を流し、溶出液を水溶出画分として得た。同様に、含水エタノール（１０％エタノール、３０％エタノール、および７０％エタノール）を４０Ｌずつ流し、溶出液をそれぞれ１０％エタノール溶出画分、３０％エタノール溶出画分、および７０％エタノール溶出画分として得た。それぞれの溶出画分は、エバポレーターおよび凍結乾燥機を用いて、乾燥体として冷蔵保存した。 (Purification of adenosine)
Adenosine was purified according to the following.
(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). HP20 was washed 3 times with ethanol and then 3 times with 50% ethanol before use. The washed HP20 was filled in a mass fractionation column and replaced with water. The same amount of distilled water was mixed with 60 L of degassed beer and flowed to the HP20 column using a medium pressure pump. A solution that passed through the HP20 column was obtained as a passing fraction. 40 L of distilled water was flowed using a medium pressure pump to obtain an eluate as a water-eluting fraction. Similarly, 40 L of hydrous ethanol (10% ethanol, 30% ethanol, and 70% ethanol) was poured, and the eluate was used as a 10% ethanol elution fraction, a 30% ethanol elution fraction, and a 70% ethanol elution fraction, respectively. Obtained. Each eluted fraction was refrigerated as a dried product using an evaporator and a lyophilizer.
（２）１０％エタノール溶出画分のＬＨ－２０分画
ＨＰ２０分画物のうち、１０％エタノール溶出画分について、Ｓｅｐｈａｄｅｘ（登録商標）ＬＨ－２０を用いて分画した（カラム用量：５００ｍＬ）。エタノールにより洗浄したＬＨ－２０を大量分画用カラムへ充填し、水により置換した。ＨＰ２０分画により得られた１０％エタノール溶出画分（２．５７ｇ）を蒸留水に溶解させ、ＬＨ－２０カラムへアプライした。１Ｌの蒸留水を流し、水溶出画分－１～５を得た。次いで、含水エタノール（３５％エタノール、７０％エタノール、および１００％エタノール）を１Ｌずつ流し、溶出液をそれぞれ３５％エタノール溶出画分、７０％エタノール溶出画分、および１００％エタノール溶出画分として得た。それぞれの溶出画分は、エバポレーターおよび凍結乾燥機を用いて、乾燥体として冷蔵保存した。 (2) LH-20 fraction of 10% ethanol-eluting fraction Of the HP20 fractions, the 10% ethanol-eluting fraction was fractionated using Sephadex® LH-20 (column dose: 500 mL). .. LH-20 washed with ethanol was filled in a mass fractionation column and replaced with water. The 10% ethanol-eluted fraction (2.57 g) obtained by the HP20 fraction was dissolved in distilled water and applied to the LH-20 column. 1 L of distilled water was poured to obtain water-eluting fractions -1 to 5. Then, 1 L of hydrous ethanol (35% ethanol, 70% ethanol, and 100% ethanol) was poured to obtain the eluate as a 35% ethanol-eluting fraction, a 70% ethanol-eluting fraction, and a 100% ethanol-eluting fraction, respectively. rice field. Each eluted fraction was refrigerated as a dried product using an evaporator and a lyophilizer.
（３）アデノシンの分離
ＬＨ－２０分画により得られた水溶出画分－３（８２．４ｍｇ）について、ＨＰＬＣ（ＣＯＳＭＯＳＩＬ　５Ｃ１８－ＰＡＱ，２０×２５０ｍｍ）を用いて、１０％エタノールにより溶出させた。次いで、５ｍｉｎから１２ｍｉｎの溶出液を濃縮し、ＨＰＬＣ（ＣＯＳＭＯＳＩＬ　５Ｃ１８－ＰＡＱ，２０×２５０ｍｍ）を用いて、エタノール－水（５：９５→１５：８５）の濃度勾配の混液により溶出させ、化合物（Ｉ）（ｔＲ＝１８．５ｍｉｎ）を得た。
化合物（Ｉ）は、ＭＳ、ＮＭＲの物理学的データの解析および標品との比較によりアデノシンであると同定した。
使用した分析機器は以下の通りである。
ＬＣ－ＭＳ；Ｑ　Ｅｘａｃｔｉｖｅ，Ｔｈｅｒｍｏ　Ｆｉｓｈｅｒ　Ｓｃｉｅｎｔｉｆｉｃ社製
ＮＭＲ；ＡＶＡＮＣＥ４００，Ｂｒｕｋｅｒ社製 (3) Separation of adenosine The water-eluted fraction-3 (82.4 mg) obtained by the LH-20 fraction was eluted with 10% ethanol using HPLC (COSMOSIL 5C18-PAQ, 20 × 250 mm). .. The eluate of 5 min to 12 min was then concentrated and eluted with a mixed solution of ethanol-water (5:95 → 15:85) using HPLC (COSMOSIL 5C18-PAQ, 20 × 250 mm) to compound (COSMOSIL 5C18-PAQ, 20 × 250 mm). I) (tR = 18.5 min) was obtained.
Compound (I) was identified as adenosine by analysis of physical data of MS and NMR and comparison with a standard.
The analytical instruments used are as follows.
LC-MS; Q Active, Thermo Fisher Scientific NMR; AVANCE400, Bruker
（４０ｋＤａタンパク質の精製）
市販のビール（１Ｌ）から下記に従い４０ｋＤａタンパク質の精製を行った。 (Purification of 40 kDa protein)
40 kDa protein was purified from commercially available beer (1 L) according to the following.
（１）陽イオン交換樹脂による分画
陽イオン交換樹脂ＳＰ　Ｓｅｐｈａｒｏｓｅ５０ｍＬを空きカラムに入れた。ビールを樹脂に吸着させた。その後、樹脂をカラムに移し替え、２０ｍＭ酢酸ナトリウム緩衝液（ｐＨ４．５）で洗浄した。次いで、２０ｍＭ酢酸ナトリウム（ｐＨ４．５）＋０．５Ｍ－ＮａＣｌで溶出し画分を集めた。得られた画分をＳＤＳ－ＰＡＧＥで評価し、４０ｋＤａのタンパク質が含まれる画分を集め陽イオン交換樹脂結合画分とした。 (1) Fractionation by cation exchange resin 50 mL of cation exchange resin SP Sepharose was placed in an empty column. Beer was adsorbed on the resin. Then, the resin was transferred to a column and washed with 20 mM sodium acetate buffer (pH 4.5). Then, it was eluted with 20 mM sodium acetate (pH 4.5) + 0.5M-NaCl and fractions were collected. The obtained fraction was evaluated by SDS-PAGE, and the fraction containing a protein of 40 kDa was collected and used as a cation exchange resin bound fraction.
（２）限外濾過（バッファー交換）
水洗浄した限外ろ過ユニット（Ｍｅｒｃｋ社製　Ａｍｉｃｏｎ　Ｕｌｔｒａ－１５　３０Ｋ）に（１）で得た陽イオン交換樹脂結合画分を１０ｍＬずつ添加し、３５００ｒｐｍで遠心し限外ろ過し濃縮液を得た。 (2) Extrafiltration (buffer exchange)
10 mL each of the cation exchange resin bonded fraction obtained in (1) was added to a water-washed ultrafiltration unit (Amicon Ultra-15 30K manufactured by Merck), and the mixture was centrifuged at 3500 rpm to obtain a concentrated solution. ..
（３）硫安分画
２０ｍＭリン酸バッファー（ｐＨ７．０）＋２Ｍ硫酸アンモニウムをビーカーに入れ、（２）で得た濃縮液を滴下、撹拌した。次に懸濁液を遠心（２３３０ｇ、１０分間、室温）した。上清を別容器に集めた。集めた溶液は限外ろ過ユニットを用いて濃縮した。濃縮液を２０ｍＭ酢酸ナトリウム（ｐＨ４．５）に加え遠心（２３３０ｇ、１０分間、室温）し、濃縮を行い、４０ｋＤａタンパク質精製品（Ｂｒａｄｆｏｒｄ定量（ウシ血清アルブミン（ＢＳＡ）換算）、２０．４ｍｇ／ｍＬ、２．２１ｍＬ）を得た。得られた４０ｋＤａタンパク質の純度はＳＤＳ－ＰＡＧＥで確認した。 (3) Ammonium sulfate fraction 20 mM phosphate buffer (pH 7.0) + 2M ammonium sulfate was placed in a beaker, and the concentrate obtained in (2) was added dropwise and stirred. The suspension was then centrifuged (2330 g, 10 minutes, room temperature). The supernatant was collected in a separate container. The collected solution was concentrated using an ultrafiltration unit. The concentrate is added to 20 mM sodium acetate (pH 4.5), centrifuged (2330 g, 10 minutes, room temperature), concentrated, and 40 kDa protein assay (Bradford quantification (bovine serum albumin (BSA) equivalent), 20.4 mg / mL. 2.21 mL) was obtained. The purity of the obtained 40 kDa protein was confirmed by SDS-PAGE.
４０ｋＤａタンパク質を酵素で消化後、ＬＣ－ＭＳ／ＭＳで分析することにより、タンパク質の同定を試みた。
ＳＤＳ－ＰＡＧＥで分離した４０ｋＤａ付近のバンドを切り出し、ジチオスレイトールによる還元（５６℃、１時間）、ヨードアセトアミドによるカルバミドメチル化（遮光下、室温、４５分間）を行った。次いで０．０１％ＰｒｏｔｅａｓｅＭａｘ含有１０ｎｇ／μＬキモトリプシン溶液（５ｍＭ塩化カルシウム、５０ｍＭ炭酸水素アンモニウム溶液）１５μＬ、５ｍＭ塩化カルシウム、５０ｍＭ炭酸水素アンモニウム溶液１５μＬを添加し一晩インキュベートした後、酵素消化液を回収した。回収した溶液を減圧乾固し、０．１％ギ酸溶液に再溶解した。
これをＬＣ－ＭＳ／ＭＳ分析に使用した。 An attempt was made to identify the protein by digesting the 40 kDa protein with an enzyme and then analyzing it by LC-MS / MS.
Bands around 40 kDa separated by SDS-PAGE were excised, reduced with dithiothreitol (56 ° C., 1 hour), and carbamid methylated with iodoacetamide (light-shielded, room temperature, 45 minutes). Then, 15 μL of 10 ng / μL chymotrypsin solution (5 mM calcium chloride, 50 mM ammonium hydrogencarbonate solution) containing 0.01% ProteinMax, 15 μL of 5 mM calcium chloride, 50 mM ammonium hydrogencarbonate solution was added, and the mixture was incubated overnight, and then the enzyme digestive juice 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.
（ＬＣ－ＭＳ／ＭＳによる測定）
ＬＣ－ＭＳ／ＭＳの測定は下記の条件で行った。
使用装置：ダイレクトフローｎａｎｏＬＣシステムＥａｓｙ－ｎＬＣ　１０００ＴＭ　（Ｔｈｅｒｍｏ　Ｓｃｉｅｎｔｉｆｉｃ）
トラップカラム：Ａｃｃｌａｉｍ　ＰｅｐＭａｐ（登録商標）（Ｔｈｅｒｍｏ　Ｓｃｉｅｎｔｉｆｉｃ）
分析カラム：ＮＡＮＯ　ＨＰＬＣ　ＣＡＰＩＬＬＡＲＹ　ＣＯＬＵＭＮ（日京テクノス（株））
液体クロマトグラフ質量分析計　Ｑ　Ｅｘａｃｔｉｖｅ　Ｐｌｕｓ（Ｔｈｅｒｍｏ　Ｓｃｉｅｎｔｉｆｉｃ）
移動相：Ａ液：０．１％ギ酸／水、Ｂ液：０．１％ギ酸／アセトニトリル
流速：３００ｎＬ／ｍｉｎ
グラジエント：０－４０％Ｂ／０－３０ｍｉｎ、４０－６０％Ｂ／３０－３５ｍｉｎ、６０－９０％Ｂ／３５－３７ｍｉｎ、９０％Ｂ／３７－４５ｍｉｎ
注入量：１０μＬ
イオン化モード：ＥＳＩ　Ｐｏｓｉｔｉｖｅ
測定範囲：ＭＳ１（ｍ／ｚ　３５０－１７５０）
Ｄａｔａ　Ｄｅｐｅｎｄｅｎｔ　Ｓｃａｎモード (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: Acclim PepMap® (Thermo Scientific)
Analytical column: NANO HPLC CAPILLARY COLUMN (Nikkyo Technos Co., Ltd.)
Liquid Chromatograph Mass Spectrometer Q Active Plus (Thermo Scientific)
Mobile phase: Liquid A: 0.1% formic acid / water, liquid B: 0.1% formic acid / acetonitrile Flow velocity: 300 nL / min
Radiant: 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）タンパク質の解析
タンパク質同定は下記の条件で行った。
検索ソフト：Ｐｒｏｔｅｏｍｅ　Ｄｉｓｃｏｖｅｒｅｒ　２．２．０．３８８（ＴｈｅｒｍｏＦｉｓｈｅｒ社製）
生物種：大麦（Ｈｏｒｄｅｕｍ　ｖｕｌｇａｒｅ）、ホップ（Ｈｕｍｕｌｕｓ）、酵母（Ｓａｃｃｈａｒｏｍｙｃｅｓ　ｃｅｒｅｖｉｓｉａｅ）
検索条件：消化酵素：Ｃｈｙｍｏｔｒｙｐｓｉｎ
プリカーサーイオン質量誤差範囲：Ｍｏｎｏｉｓｏｔｏｐｉｃ、±１０ｐｐｍ
プロダクトイオン質量誤差範囲：±０．０２Ｄａ
最大ミスクリベージ数：５
コンフィデンスレベル（Ｐｅｒｃｏｌａｔｏｒ）：Ｈｉｇｈ（確からしさ３段階のうち最も確率が高いレベル）
データベース：ＳｗｉｓｓＰｒｏｔ (4) Protein analysis Protein identification was performed under the following conditions.
Search software: Proteome Discoverer 2.2.0.3888 (manufactured by Thermo Fisher)
Species: Barley (Hordeum vulgare), Hops (Humulus), Yeast (Saccharomyces cerevisiae)
Search condition: Digestive enzyme: Chymotrypsin
Precursor ion mass error range: Monoisotopic, ± 10 ppm
Product ion mass error range: ± 0.02 Da
Maximum number of missed cribes: 5
Confidence level (Percolator): High (the most probable level of the three levels of certainty)
Database: SwissProt
その結果、４０ｋＤａタンパク質は大麦由来Ｓｅｒｐｉｎ　Ｚ４（配列カバー率：７７．２％）及び大麦由来Ｓｅｒｐｉｎ　Ｚ７（配列カバー率：７２．８％）であることがわかった。 As a result, it was found that the 40 kDa protein was barley-derived Serpin Z4 (sequence coverage: 77.2%) and barley-derived Serpin Z7 (sequence coverage: 72.8%).
（市販のビールテイストアルコール飲料にアデノシンを添加した場合の官能評価）
市販のビールテイストアルコール飲料に、アデノシンを添加し、ふくらみの官能評価を行った。
当該ビールテイストアルコール飲料は、原料に麦芽を含み、原料中の麦芽の比率が５０重量％未満であり、プリン体の濃度が５ｍｇ／１００ｍＬ未満のビールテイストアルコール飲料である。
原材料は発泡酒、麦芽、ホップ、糖類、食物繊維、スピリッツ（小麦）であり、栄養成分として１００ｍｌあたりアルコール分４％、タンパク質０～０．２ｇ、糖質０．５～０．８ｇ、プリン体約２．０ｍｇを含む。 (Sensory evaluation when adenosine is added to a commercially available beer-taste alcoholic beverage)
Adenosine was added to a commercially available beer-taste alcoholic beverage, and the sensory evaluation of swelling was performed.
The beer-taste alcoholic beverage is a beer-taste alcoholic beverage in which malt is contained in the raw material, the ratio of malt in the raw material is less than 50% by weight, and the concentration of purine is less than 5 mg / 100 mL.
The raw materials are low-malt beer, malt, hops, sugars, dietary fiber, and spirits (wheat). As nutritional components, alcohol content is 4%, protein is 0 to 0.2 g, sugar is 0.5 to 0.8 g, and purine is used. Contains about 2.0 mg.
ふくらみの官能評価の基準点は以下の通りである。
専門パネル５名により０．０５点刻みで下記の基準によりスコア化、そのスコア値を平均化した。
ふくらみ強度は以下の基準である。
０点：全く感じられない
１点：やや感じられる
２点：明確に感じる
３点：非常に感じる
基準点として、評価対象とする上記市販のビールテイストアルコール飲料と同じ上記市販のビールテイストアルコール飲料を基準のビールテイストアルコール飲料（Ｉ）としてそのふくらみを０．７点とした。また、他の市販のビールテイストアルコール飲料を基準のビールテイストアルコール飲料（ＩＩ）としてそのふくらみを基準点の１．５点とした。
当該基準のビールテイストアルコール飲料（ＩＩ）は原料中の麦芽の比率が５０重量％以上であるビールテイストアルコール飲料である。
原材料は麦芽、ホップであり、栄養成分として１００ｍｌあたりアルコール分５．５％、タンパク質０．４～０．６ｇ、糖質３．６ｇ、プリン体約１２．５ｍｇを含む。 The reference points for the sensory evaluation of the bulge are as follows.
Five expert panels scored in increments of 0.05 points according to the following criteria, and the score values were averaged.
The bulge strength is based on the following criteria.
0 points: Not felt at all 1 point: Slightly felt 2 points: Clearly felt 3 points: As a very felt reference point, the same commercially available beer-taste alcoholic beverage as the above-mentioned commercially available beer-taste alcoholic beverage to be evaluated is used. The bulge was given as 0.7 points as the standard beer-taste alcoholic beverage (I). Further, another commercially available beer-taste alcoholic beverage was used as the standard beer-taste alcoholic beverage (II), and its bulge was set to 1.5 points as the reference point.
The standard beer-taste alcoholic beverage (II) is a beer-taste alcoholic beverage in which the ratio of malt in the raw material is 50% by weight or more.
The raw materials are malt and hops, and the nutritional components include 5.5% alcohol, 0.4 to 0.6 g of protein, 3.6 g of sugar, and about 12.5 mg of purines per 100 ml.
また、ビールらしい苦味の官能評価の基準点は以下の通りである。
専門パネル４名により０．０５点刻みで下記の基準によりスコア化、そのスコア値を平均化した。
ビールらしい苦味強度は以下の基準である。
０点：全く感じられない
１点：やや感じられる
２点：明確に感じる
３点：非常に感じる
基準点として、評価対象とする上記市販のビールテイストアルコール飲料と同じ上記市販のビールテイストアルコール飲料を基準のビールテイストアルコール飲料（Ｉ）としてそのビールらしい苦味を１．０点とした。また、上述の他の市販のビールテイストアルコール飲料を基準のビールテイストアルコール飲料（ＩＩ）としてそのビールらしい苦味を基準点の２．０点とした。 The reference points for sensory evaluation of beer-like bitterness are as follows.
Four specialized panels scored in increments of 0.05 points according to the following criteria, and the score values were averaged.
Beer-like bitterness intensity is based on the following criteria.
0 points: Not felt at all 1 point: Slightly felt 2 points: Clearly felt 3 points: As a reference point that is very felt, the same commercially available beer-taste alcoholic beverage as the above-mentioned commercially available beer-taste alcoholic beverage to be evaluated is used. As the standard beer taste alcoholic beverage (I), the beer-like bitterness was given as 1.0 point. Further, the above-mentioned other commercially available beer-taste alcoholic beverages were used as the standard beer-taste alcoholic beverages (II), and the beer-like bitterness was set to 2.0 points as the reference point.
官能評価の手順は以下の通りである。
（１）ビールテイストアルコール飲料を最終容量の１／１０容量（ｖ／ｖ）バイアル瓶に分注する
（２）アデノシンを任意の重量で秤量して加える
（３）３０秒ソニケーションする
（４）３０分室温で静置する
（５）ビールテイストアルコール飲料を最終容量にフィルアップする
（６）分注して飲み込み評価する The procedure for sensory evaluation is as follows.
(1) Dispense beer-taste alcoholic beverages into 1/10 volume (v / v) vials of the final volume (2) Weigh and add adenosine to any weight (3) Sonic for 30 seconds (4) Let stand at room temperature for 30 minutes (5) Fill up the beer-taste alcoholic beverage to the final volume (6) Dispense and evaluate by swallowing
（市販のビールテイストアルコール飲料の分析）
官能評価に使用した市販のビールテイストアルコール飲料に含まれるアデノシンの濃度を以下の手順によりＬＣ－ＭＳで定量した。
（１）標品の調製及び検量線の作製
アデノシンについて、それぞれ下記の濃度となるように希釈し、０．２２μｍのフィルターに通してから測定に供した。
最終濃度：０．００１ｐｐｍ，０．０２５ｐｐｍ，０．０５０ｐｐｍ，０．１００ｐｐｍ，０．２００ｐｐｍ，０．３００ｐｐｍ，０．５００ｐｐｍ，０．７５０ｐｐｍ，１．０００ｐｐｍ
（１ｐｐｍ＝１μｇ／ｍＬである）
希釈液には、５％（ｖ／ｖ）のエタノール水溶液を用いた。
なお、標品の分析結果において、検量線の直線性が保たれる範囲（Ｒ２＞０．９９）に測定値が入るような希釈倍率の測定値を採用した。 (Analysis of commercial beer-taste alcoholic beverages)
The concentration of adenosine contained in the commercially available beer-taste alcoholic beverage used for the sensory evaluation was quantified by LC-MS by the following procedure.
(1) Preparation of standard and preparation of calibration curve Adenosine was diluted to the following concentrations, passed through a 0.22 μm filter, and then subjected to 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.
Note that in the analysis of a preparation was employed measurements of the dilution ratio as the measured value is in the range (R 2> 0.99) the linearity of the calibration curve is maintained.
ＬＣ－ＭＳの測定条件は以下の通りである。
ＬＣ－ＭＳ：エービー・サイエックス社製Ｘ５００Ｒ
分離カラム：Ｗａｔｅｒｓ社製　ＨＳＳＴ３　１．８μｍ，２．１ｘ１５０ｍｍ
溶離液：
Ａ液：０．１％ギ酸／水、Ｂ液：０．１％ギ酸／アセトニトリル
グラジエント：Ａ液：Ｂ液＝９８：２→２：９８（２７ｍｉｎ）
注入量：５μＬ
流速：０．２ｍＬ／ｍｉｎ
カラムオーブン：４０℃
（ＭＳ）
イオン化モード：ＥＳＩ　Ｐｏｓｉｔｉｖｅ
測定範囲：ＭＳ１（ｍ／ｚ　１００－１０００）
Ｄａｔａ　Ｉｎｄｅｐｅｎｄｅｎｔ　Ｓｃａｎモード
イオン源温度：３５０℃ The measurement conditions for LC-MS are as follows.
LC-MS: X500R manufactured by AB SIX Co., Ltd.
Separation column: Waters HSST3 1.8 μm, 2.1x150 mm
Eluent:
Liquid A: 0.1% formic acid / water, liquid B: 0.1% formic acid / acetonitrile gradient: liquid A: liquid 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
（２）市販のビールテイストアルコール飲料からの測定用試料の調製
市販のビールテイストアルコール飲料をソニケーションにより脱気し、気泡が落ち着いてから適宜希釈し、０．２２μｍのフィルターに通してから測定に供した。
希釈液には、５％（ｖ／ｖ）のエタノール水溶液を用いた。
市販のビールテイストアルコール飲料に含まれる各アデノシンの濃度をコントロールとした。 (2) Preparation of measurement sample from commercially available beer-taste alcoholic beverages Commercially available beer-taste alcoholic beverages are degassed by sonication, diluted appropriately after the bubbles have settled, passed through a 0.22 μm filter, and then measured. Served.
A 5% (v / v) aqueous ethanol solution was used as the diluent.
The concentration of each adenosine contained in a commercially available beer-taste alcoholic beverage was used as a control.
（実施例１：本発明の第一形態におけるアデノシンおよび／または４０ｋＤａタンパク質添加によるふくらみの評価）
市販のビールテイストアルコール飲料（コントロール）に含まれるアデノシンの濃度は４．２５ｐｐｍであった。これに対してアデノシン濃度が５ｐｐｍ、８ｐｐｍにそれぞれなるようにアデノシンを加えて官能評価を行った（試料１及び試料２）。
また、市販のビールテイストアルコール飲料（コントロール）に含まれる４０ｋＤａタンパク質の濃度は０ｐｐｍであった。これに対してアデノシン濃度が５ｐｐｍ、４０ｋＤａタンパク質濃度が５ｐｐｍ、９ｐｐｍにそれぞれなるようにアデノシン及び４０ｋＤａタンパク質を加えて官能評価を行った（試料３及び試料４）。
また、市販のビールテイストアルコール飲料（コントロール）に含まれる４０ｋＤａタンパク質濃度が５ｐｐｍ、９ｐｐｍ、２５ｐｐｍになるように４０ｋＤａタンパク質を加えて官能評価を行った（対比試料１、対比試料２及び対比試料３）。
４０ｋＤａタンパク質には、上記で精製したものを使用した。
官能評価の結果を表１に示した。
また、各試料のプリン体濃度を下記表１に示す。本明細書における「プリン体濃度」は、上述の通りアデニン、グアニン、キサンチン、ヒポキサンチンのプリン体塩基４種の総量であり、例えば、過塩素酸による加水分解後にＬＣ－ＭＳ／ＭＳ（液体クロマトグラフィー質量分析法）を用いて検出することができる。 (Example 1: Evaluation of swelling by addition of adenosine and / or 40 kDa protein in the first embodiment of the present invention)
The concentration of adenosine contained in the commercially available beer-taste alcoholic beverage (control) was 4.25 ppm. On the other hand, adenosine was added so that the adenosine concentrations were 5 ppm and 8 ppm, respectively, and sensory evaluation was performed (Sample 1 and Sample 2).
The concentration of 40 kDa protein contained in a commercially available beer-taste alcoholic beverage (control) was 0 ppm. On the other hand, sensory evaluation was performed by adding adenosine and 40 kDa protein so that the adenosine concentration was 5 ppm and the 40 kDa protein concentration was 5 ppm and 9 ppm, respectively (Sample 3 and Sample 4).
In addition, 40 kDa protein was added so that the 40 kDa protein concentration contained in the commercially available beer-taste alcoholic beverage (control) was 5 ppm, 9 ppm, and 25 ppm, and sensory evaluation was performed (comparison sample 1, contrast sample 2 and contrast sample 3). ..
The 40 kDa protein used was the one purified above.
The results of the sensory evaluation are shown in Table 1.
The purine concentration of each sample is shown in Table 1 below. As described above, the "purine concentration" in the present specification is the total amount of four purine bases of adenine, guanine, xanthine, and hypoxanthine, and is, for example, LC-MS / MS (liquid chromatograph) after hydrolysis with perchloric acid. It can be detected by using a graph mass spectrometry method).
Figure JPOXMLDOC01-appb-T000001
Figure JPOXMLDOC01-appb-T000001
表１に示す結果から、原料中の麦芽の比率が５０重量％未満であり、プリン体の濃度が５ｍｇ／１００ｍＬ未満であるビールテイストアルコール飲料につき、アデノシン濃度が５ｐｐｍ以上（試料１（５ｐｐｍ）、試料２（８ｐｐｍ））であるとふくらみが増強されることが分かる。
また、市販のビールテイストアルコール飲料に、アデノシン及び４０ｋＤａタンパク質を加えることにより、ふくらみをより増強できることが分かる（試料３及び試料４）。
対比試料１、対比試料２及び対比試料３の結果から、４０ｋＤａタンパク質を添加するだけでもふくらみを増強することができることがわかる。しかしながら、上記試料１における官能評価のコントロールからの増加値（０．０３）と対比試料１における官能評価のコントロールからの増加値（０．０５）の和で求められる、アデノシン（５ｐｐｍ）と４０ｋＤａタンパク質（５ｐｐｍ）を併用したときの相加効果（０．０８）よりも、試料３における官能評価のコントロールからの増加値（０．１３）が大きいことから、アデノシンと４０ｋＤａタンパク質を併用することにより、予想できない相乗効果が発揮されているといえる。
また、上記試料１における官能評価のコントロールからの増加値（０．０３）と対比試料２における官能評価のコントロールからの増加値（０．０８）の和で求められる、アデノシン（５ｐｐｍ）と４０ｋＤａタンパク質（９ｐｐｍ）を併用したときの相加効果（０．１１）よりも、試料４における官能評価のコントロールからの増加値（０．１７）が大きいことからも、アデノシンと４０ｋＤａタンパク質を併用することにより、予想できない相乗効果が発揮されているといえる。 From the results shown in Table 1, for a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight and the concentration of purine is less than 5 mg / 100 mL, the adenosine concentration is 5 ppm or more (Sample 1 (5 ppm), It can be seen that the swelling is enhanced in the case of sample 2 (8 ppm)).
It is also found that the swelling can be further enhanced by adding adenosine and 40 kDa protein to a commercially available beer-taste alcoholic beverage (Sample 3 and Sample 4).
From the results of the comparison sample 1, the comparison sample 2 and the comparison sample 3, it can be seen that the swelling can be enhanced only by adding the 40 kDa protein. However, the adenosine (5 ppm) and 40 kDa protein obtained by the sum of the increase value (0.03) from the control of the sensory evaluation in the sample 1 and the increase value (0.05) from the control of the sensory evaluation in the comparison sample 1. Since the increase value (0.13) from the control of the sensory evaluation in sample 3 is larger than the additive effect (0.08) when (5 ppm) is used in combination, by using adenosine and 40 kDa protein in combination, It can be said that an unpredictable synergistic effect is exhibited.
Further, adenosine (5 ppm) and 40 kDa protein obtained by the sum of the increase value (0.03) from the control of the sensory evaluation in the sample 1 and the increase value (0.08) from the control of the sensory evaluation in the comparison sample 2. Since the increase value (0.17) from the control of the sensory evaluation in sample 4 is larger than the additive effect (0.11) when (9 ppm) is used in combination, the combined use of adenosine and 40 kDa protein It can be said that an unexpected synergistic effect is exhibited.
（実施例２：本発明の第二形態におけるアデノシン添加によるふくらみの評価）
市販のビールテイストアルコール飲料（コントロール）に含まれるアデノシンの濃度は４．２５ｐｐｍであった。これに対してアデノシン濃度が８ｐｐｍ、１２ｐｐｍ、１８ｐｐｍ、２３ｐｐｍにそれぞれなるようにアデノシンを加えてふくらみの官能評価を行った（対比試料４、試料５、試料６及び対比試料５）。
官能評価の結果を表２に示した。
また、各試料のプリン体濃度を下記表２に示す。本明細書における「プリン体濃度」は、上述の通りアデニン、グアニン、キサンチン、ヒポキサンチンのプリン体塩基４種の総量であり、例えば、過塩素酸による加水分解後にＬＣ－ＭＳ／ＭＳ（液体クロマトグラフィー質量分析法）を用いて検出することができる。 (Example 2: Evaluation of swelling due to addition of adenosine in the second embodiment of the present invention)
The concentration of adenosine contained in the commercially available beer-taste alcoholic beverage (control) was 4.25 ppm. On the other hand, adenosine was added so that the adenosine concentrations were 8 ppm, 12 ppm, 18 ppm, and 23 ppm, respectively, and the sensory evaluation of the swelling was performed (comparison sample 4, sample 5, sample 6 and contrast sample 5).
The results of the sensory evaluation are shown in Table 2.
The purine concentration of each sample is shown in Table 2 below. As described above, the "purine concentration" in the present specification is the total amount of four purine bases of adenine, guanine, xanthine, and hypoxanthine, and is, for example, LC-MS / MS (liquid chromatograph) after hydrolysis with perchloric acid. It can be detected by using a graph mass spectrometry method).
Figure JPOXMLDOC01-appb-T000002
Figure JPOXMLDOC01-appb-T000002
表２に示す結果から、原料中の麦芽の比率が５０重量％未満であるビールテイストアルコール飲料であって、プリン体の濃度が５ｍｇ／１００ｍＬ未満である上記飲料につき、アデノシン濃度が１２ｐｐｍ以上（試料５、試料６、対比試料５）であると、コントロールのふくらみの評価に対し０．１点以上増加し、効果的にふくらみが増強されることが分かる。
一方で、上記表２の試料６及び対比試料５を比較すると、アデノシン濃度が１８ｐｐｍを超えても濃度増加分に応じてふくらみが効果的に増加していないことが確認される。
よって、原料中の麦芽の比率が５０重量％未満であるビールテイストアルコール飲料であって、プリン体の濃度が５ｍｇ／１００ｍＬ未満である上記飲料につき、アデノシン濃度が１２～１８ｐｐｍの範囲において、効果的にふくらみが増強されることが分る。 From the results shown in Table 2, the adenosine concentration is 12 ppm or more (sample) for the beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight and the purine concentration is less than 5 mg / 100 mL. In the case of 5, sample 6 and comparison sample 5), the bulge is increased by 0.1 points or more with respect to the evaluation of the bulge of the control, and it can be seen that the bulge is effectively enhanced.
On the other hand, when the sample 6 and the contrast sample 5 in Table 2 above are compared, it is confirmed that the swelling does not effectively increase according to the increase in the concentration even if the adenosine concentration exceeds 18 ppm.
Therefore, it is effective in the range of adenosine concentration of 12 to 18 ppm for the beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight and the concentration of purine is less than 5 mg / 100 mL. It can be seen that the swelling is enhanced.
（実施例３：本発明の第二形態におけるアデノシン及び４０ｋＤａタンパク質の添加によるふくらみの評価）
上記市販のビールテイストアルコール飲料に含まれるアデノシンの濃度は４．２５ｐｐｍであり、４０ｋＤａタンパク質の濃度は０ｐｐｍであった。
これに対してアデノシン濃度が８ｐｐｍ、１２ｐｐｍ、１８ｐｐｍにそれぞれなるようにアデノシンを加え、４０ｋＤａタンパク質濃度が５ｐｐｍになるように４０ｋＤａタンパク質を加えてふくらみの官能評価を行った（対比試料８、試料７及び試料８）。
４０ｋＤａタンパク質には、上記で精製したものを使用した。
なお、対比のため、上記市販のビールテイストアルコール飲料に対して４０ｋＤａタンパク質濃度が５ｐｐｍ、２５ｐｐｍになるように４０ｋＤａタンパク質だけを添加してふくらみの官能評価を行った（対比試料６及び対比試料７）。
官能評価の結果を表３に示した。また、各試料のプリン体濃度を下記表３に示す。 (Example 3: Evaluation of swelling due to addition of adenosine and 40 kDa protein in the second embodiment of the present invention)
The concentration of adenosine contained in the above-mentioned commercially available beer-taste alcoholic beverage was 4.25 ppm, and the concentration of 40 kDa protein was 0 ppm.
On the other hand, adenosine was added so that the adenosine concentration was 8 ppm, 12 ppm, and 18 ppm, respectively, and 40 kDa protein was added so that the 40 kDa protein concentration was 5 ppm, and the sensory evaluation of the swelling was performed (contrast sample 8, sample 7, and). Sample 8).
The 40 kDa protein used was the one purified above.
For comparison, only 40 kDa protein was added to the commercially available beer-taste alcoholic beverage so that the 40 kDa protein concentration was 5 ppm and 25 ppm, and the sensory evaluation of the swelling was performed (contrast sample 6 and contrast sample 7). ..
The results of the sensory evaluation are shown in Table 3. The purine concentration of each sample is shown in Table 3 below.
Figure JPOXMLDOC01-appb-T000003
Figure JPOXMLDOC01-appb-T000003
上記表３の結果から、市販のビールテイストアルコール飲料に、アデノシン及び４０ｋＤａタンパク質（５ｐｐｍ）を加えることにより、ふくらみをより増強できることが分かる（対比試料８、試料７及び試料８）。
また、４０ｋＤａタンパク質を添加するだけでもふくらみを増強することができることがわかる（対比試料６及び対比試料７）。しかしながら、上記試料５における官能評価のコントロールからの増加値（０．１１）と対比試料６における官能評価のコントロールからの増加値（０．０６）の和で求められる、アデノシン（１２ｐｐｍ）と４０ｋＤａタンパク質（５ｐｐｍ）を併用したときの相加効果（０．１７）よりも、試料７における官能評価のコントロールからの増加値（０．２３）が大きいことから、アデノシンと４０ｋＤａタンパク質を併用することにより、予想できない相乗効果が発揮されているといえる。
上記相乗効果は、上記試料６における官能評価のコントロールからの増加値（０．１４）と対比試料６における官能評価のコントロールからの増加値（０．０６）の和で求められる、アデノシン（１８ｐｐｍ）と４０ｋＤａタンパク質（５ｐｐｍ）を併用したときの相加効果（０．２）よりも、試料８における官能評価のコントロールからの増加値（０．２５）が大きいことからも確認できる。 From the results in Table 3 above, it can be seen that the swelling can be further enhanced by adding adenosine and 40 kDa protein (5 ppm) to a commercially available beer-taste alcoholic beverage (Comparative Sample 8, Sample 7, and Sample 8).
Further, it can be seen that the swelling can be enhanced only by adding the 40 kDa protein (contrast sample 6 and contrast sample 7). However, the adenosine (12 ppm) and 40 kDa protein obtained by the sum of the increase value (0.11) from the sensory evaluation control in the sample 5 and the increase value (0.06) from the sensory evaluation control in the contrast sample 6 are obtained. Since the increase value (0.23) from the control of the sensory evaluation in sample 7 is larger than the additive effect (0.17) when (5 ppm) is used in combination, by using adenosine and 40 kDa protein in combination, It can be said that an unpredictable synergistic effect is exhibited.
The synergistic effect is determined by the sum of the increase value (0.14) from the sensory evaluation control in the sample 6 and the increase value (0.06) from the sensory evaluation control in the comparison sample 6, adenosine (18 ppm). It can also be confirmed from the fact that the increase value (0.25) from the control of the sensory evaluation in the sample 8 is larger than the additive effect (0.2) when the 40 kDa protein (5 ppm) is used in combination.
（実施例４：本発明の第二形態におけるアデノシン及び／又は４０ｋＤａタンパク添加によるビールらしい苦味の評価）
市販のビールテイストアルコール飲料（コントロール）に含まれるアデノシンの濃度は４．２５ｐｐｍであり、４０ｋＤａタンパクの濃度は０ｐｐｍであった。
これに対してアデノシン濃度が８ｐｐｍ、１２ｐｐｍ、１８ｐｐｍ、２３ｐｐｍにそれぞれなるようにアデノシンを加えてビールらしい苦みの官能評価を行った（対比試料４、試料５、試料６及び対比試料５）。
また、市販のビールテイストアルコール飲料に対してアデノシン濃度が１２ｐｐｍ、１８ｐｐｍ、４０ｋＤａタンパク質濃度が５ｐｐｍにそれぞれなるように上記飲料に対してアデノシン及び４０ｋＤａタンパクを加えてビールらしい苦味の官能評価を行った（試料７、試料８）。
４０ｋＤａタンパク質には、上記で精製したものを使用した。
また、対比のため、市販のビールテイストアルコール飲料に含まれる４０ｋＤａタンパクだけを添加してのビールらしい苦みの官能評価を行った（対比試料６）。
官能評価の結果を表４に示した。また、各試料のプリン体濃度を下記表４に示す。 (Example 4: Evaluation of beer-like bitterness by adding adenosine and / or 40 kDa protein in the second embodiment of the present invention)
The concentration of adenosine contained in the commercially available beer-taste alcoholic beverage (control) was 4.25 ppm, and the concentration of 40 kDa protein was 0 ppm.
On the other hand, adenosine was added so that the adenosine concentrations were 8 ppm, 12 ppm, 18 ppm, and 23 ppm, respectively, and the sensory evaluation of beer-like bitterness was performed (comparison sample 4, sample 5, sample 6 and contrast sample 5).
In addition, adenosine and 40 kDa protein were added to the above-mentioned beverage so that the adenosine concentration was 12 ppm, 18 ppm, and the 40 kDa protein concentration were 5 ppm, respectively, with respect to the commercially available beer-taste alcoholic beverage, and the sensory evaluation of the bitterness like beer was performed (). Sample 7, sample 8).
The 40 kDa protein used was the one purified above.
For comparison, a sensory evaluation of beer-like bitterness was performed by adding only 40 kDa protein contained in a commercially available beer-taste alcoholic beverage (contrast sample 6).
The results of the sensory evaluation are shown in Table 4. The purine concentration of each sample is shown in Table 4 below.
Figure JPOXMLDOC01-appb-T000004
Figure JPOXMLDOC01-appb-T000004
表４に示す結果から、原料中の麦芽の比率が５０重量％未満であるビールテイストアルコール飲料であって、プリン体の濃度が５ｍｇ／１００ｍＬ未満である上記飲料につき、アデノシン濃度が増加すると、ビールらしい苦味が増加することが分る。そして、アデノシン濃度が１２ｐｐｍ以上（試料５、試料６、対比試料５）であると、コントロールのビールらしい苦味の評価に対し０．１点以上増加し、効果的にビールらしい苦味が増強されることが分かる。
また、市販のビールテイストアルコール飲料に、アデノシン及び４０ｋＤａタンパク質（５ｐｐｍ）を加えることにより、ビールらしい苦味をより増強できることが分かる（試料７及び試料８）。
また、４０ｋＤａタンパク質を添加するだけでもビールらしい苦味を増強することができることがわかる（対比試料６）。しかしながら、上記試料５における官能評価のコントロールからの増加値（０．１９）と対比試料６における官能評価のコントロールからの増加値（０．０４）の和で求められる、アデノシン（１２ｐｐｍ）と４０ｋＤａタンパク質（５ｐｐｍ）を併用したときの相加効果（０．２３）よりも、試料７における官能評価のコントロールからの増加値（０．４）が大きいことから、アデノシンと４０ｋＤａタンパク質を併用することにより、予想できない相乗効果が発揮されているといえる。
上記相乗効果は、上記試料６における官能評価のコントロールからの増加値（０．３４）と対比試料６における官能評価のコントロールからの増加値（０．０４）の和で求められる、アデノシン（１８ｐｐｍ）と４０ｋＤａタンパク質（５ｐｐｍ）を併用したときの相加効果（０．３８）よりも、試料８における官能評価のコントロールからの増加値（０．５８）が大きいことからも確認できる。
なお、原料中の麦芽の比率が５０重量％未満であるビールテイストアルコール飲料であって、プリン体の濃度が５ｍｇ／１００ｍＬ未満である上記飲料につき、アデノシン濃度が１８ｐｐｍを超えると、後味の苦み又は収斂味を感じるパネラーがいたことから、上記ビールテイストアルコール飲料につき、後味の苦味又は収斂味を増加させることなく、ふくらみ及びビールらしい苦味を増強するには、アデノシンの濃度が１２～１８ｐｐｍが好ましいと考えられる。 From the results shown in Table 4, when the adenosine concentration is increased in the beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight and the purine concentration is less than 5 mg / 100 mL, the beer is beer. It can be seen that the peculiar bitterness increases. When the adenosine concentration is 12 ppm or more (Sample 5, Sample 6, Comparative Sample 5), the beer-like bitterness of the control is increased by 0.1 points or more with respect to the evaluation of the beer-like bitterness, and the beer-like bitterness is effectively enhanced. I understand.
Further, it can be seen that the bitterness like beer can be further enhanced by adding adenosine and 40 kDa protein (5 ppm) to a commercially available beer-taste alcoholic beverage (Sample 7 and Sample 8).
Further, it can be seen that the bitterness like beer can be enhanced only by adding 40 kDa protein (contrast sample 6). However, the adenosine (12 ppm) and 40 kDa protein obtained by the sum of the increase value (0.19) from the sensory evaluation control in the sample 5 and the increase value (0.04) from the sensory evaluation control in the comparison sample 6. Since the increase value (0.4) from the control of the sensory evaluation in sample 7 is larger than the additive effect (0.23) when (5 ppm) is used in combination, by using adenosine and 40 kDa protein in combination, It can be said that an unpredictable synergistic effect is exhibited.
The synergistic effect is determined by the sum of the increase value (0.34) from the sensory evaluation control in the sample 6 and the increase value (0.04) from the sensory evaluation control in the comparison sample 6, adenosine (18 ppm). It can also be confirmed from the fact that the increase value (0.58) from the control of the sensory evaluation in the sample 8 is larger than the additive effect (0.38) when the 40 kDa protein (5 ppm) is used in combination.
For a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight and the concentration of purine is less than 5 mg / 100 mL, if the adenosine concentration exceeds 18 ppm, the aftertaste becomes bitter or Since some panelists felt astringent taste, it is preferable that the concentration of adenosine is 12 to 18 ppm in order to enhance the swelling and beer-like bitterness of the above beer-taste alcoholic beverage without increasing the bitterness or astringent taste of the aftertaste. Conceivable.
以上の結果から、原料中の麦芽の比率が５０重量％未満であり、プリン体の濃度が５ｍｇ／１００ｍＬ未満であるビールテイストアルコール飲料におけるアデノシン濃度を１２～１８ｐｐｍとすることにより、効果的にふくらみ及びビールらしい苦味を増強できることを確認した。さらに、アデノシンと４０ｋＤａタンパク質を併用することにより、ふくらみ及びビールらしい苦味の増強において予想できない相乗効果が発揮されているといえる。 From the above results, effective swelling is achieved by setting the adenosine concentration in a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight and the concentration of purine is less than 5 mg / 100 mL to 12 to 18 ppm. And it was confirmed that the bitterness like beer can be enhanced. Furthermore, it can be said that the combined use of adenosine and 40 kDa protein exerts an unexpected synergistic effect in enhancing swelling and beer-like bitterness.
本発明によれば、原料中の麦芽の比率が５０重量％未満であるビールテイストアルコール飲料において、ふくらみが増強された飲料を提供することができる。また、本発明によれば、原料中の麦芽の比率が５０重量％未満であり、プリン体の濃度が５ｍｇ／１００ｍＬ未満であるビールテイストアルコール飲料において、ふくらみ及びビールらしい苦味が増強された飲料を提供することができる。


  According to the present invention, it is possible to provide a beverage with enhanced swelling in a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight. Further, according to the present invention, in a beer-taste alcoholic beverage in which the ratio of malt in the raw material is less than 50% by weight and the concentration of purines is less than 5 mg / 100 mL, a beverage having enhanced swelling and beer-like bitterness is produced. Can be provided.

Claims (4)

  1. 原料中の麦芽の比率が５０重量％未満であるビールテイストアルコール飲料であって、
    プリン体の濃度が５ｍｇ／１００ｍＬ未満、アデノシンの濃度が５～１０ｐｐｍであるビールテイストアルコール飲料。     A beer-taste alcoholic beverage in which the proportion of malt in the raw material is less than 50% by weight.
    A beer-taste alcoholic beverage with a purine concentration of less than 5 mg / 100 mL and an adenosine concentration of 5-10 ppm.
  2. 原料中の麦芽の比率が５０重量％未満であるビールテイストアルコール飲料であって、
    プリン体の濃度が５ｍｇ／１００ｍＬ未満、アデノシンの濃度が１２～１８ｐｐｍであるビールテイストアルコール飲料。     A beer-taste alcoholic beverage in which the proportion of malt in the raw material is less than 50% by weight.
    A beer-taste alcoholic beverage with a purine concentration of less than 5 mg / 100 mL and an adenosine concentration of 12-18 ppm.
  3. さらに、分子量３５～５０ｋＤａのタンパク質を含み、
    前記タンパク質の濃度が１ｐｐｍ以上である請求項１又は２に記載のビールテイストアルコール飲料。     In addition, it contains a protein with a molecular weight of 35-50 kDa,
    The beer-taste alcoholic beverage according to claim 1 or 2, wherein the protein concentration is 1 ppm or more.
  4. 前記タンパク質の濃度が３０ｐｐｍ以下である請求項３に記載のビールテイストアルコール飲料。

          The beer-taste alcoholic beverage according to claim 3, wherein the protein concentration is 30 ppm or less.
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JP2014079220A (en) * 2012-10-18 2014-05-08 Asahi Breweries Ltd Manufacturing method of fermented malt beverage inducing enhanced feeling of stimulation
JP2014117205A (en) * 2012-12-14 2014-06-30 Kirin Brewery Co Ltd Production method of low-purine body effervescent alcoholic beverage by using corn grits as main raw material
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