Beer-flavored beverage and method for producing beer-flavored beverage

Abstract

To provide a beer-flavored beverage having an excellent taste and little astringency, while having a reduced content of purines. [Solution] A beer-flavored beverage having a purine content of 15 ppm or less and a peptide content of 35 to 50 kDa molecular weight of 1.5 to 50 ppm.

Description

[Technical Field]

[0001] The present invention relates to a beer-taste beverage and a method for producing a beer-taste beverage. [Background technology]

[0002] In recent years, growing health consciousness has led to increased demand for beer-flavored beverages with reduced or zero purines. Purines are a general term for compounds with a purine skeleton. Purines contained in beer-flavored beverages typically include eight types: nucleosides such as adenosine, guanosine, inosine, and xanthosine, and purine bases such as adenine, guanine, hypoxanthine, and xanthine. The purine content of beer-flavored beverages tends to increase with a higher malt content. Therefore, conventional low-purine or zero-purine beer-flavored beverages reduce their purine content by keeping the malt content low and, if necessary, further performing processes such as purine adsorption and dilution. For example, Patent Document 1 describes a method for producing a beer-flavored beverage with a purine content of 0.50 mg / 100 mL or less, in which the malt content is kept below 5% by weight and the purine content is reduced by activated carbon treatment. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Japanese Patent Publication No. 2021-168688 Summary of the Invention [Problem to be solved by the invention]

[0004] However, conventional purine-free or purine-free beer-flavored beverages have an unsatisfactory taste. For example, while it is possible to reduce the purine content of a beer-flavored beverage by adsorbing and removing the purines using an adsorbent such as activated carbon, adsorbing and removing most of the purines contained in the beer-flavored beverage can result in a poor taste. Similarly, reducing the purine content of a beer-flavored beverage by dilution can also result in a weaker, less palatable taste. Therefore, there has been a demand for a beer-flavored beverage that has a reduced purine content but still has an excellent taste.

[0005] The present inventors conducted extensive research focusing on the taste of beer-taste beverages and found that beer-taste beverages containing at least a certain amount of peptides with a molecular weight of 35 to 50 kDa have excellent taste. However, the present inventors also found that if the content of peptides with a molecular weight of 35 to 50 kDa in a beer-taste beverage exceeds a certain amount, the beer-taste beverage will have an undesirably strong astringent taste. Furthermore, there have been no previous cases in which the appropriate range of peptide content with a molecular weight of 35 to 50 kDa has been specifically verified in terms of the balance between taste and astringency in beer-flavored beverages with reduced purine content.

[0006] The present invention aims to provide a beer-taste beverage that has a reduced purine content, yet has an excellent taste and little astringency, and a method for producing the same. [Means for solving the problem]

[0007] The present inventors conducted extensive research into the effects of the content of peptides with molecular weights of 35 to 50 kDa on the taste and astringency of beer-taste beverages with reduced purine content, and discovered that beer-taste beverages with a content of peptides with molecular weights of 35 to 50 kDa within a specific range have excellent taste and little astringency, leading to the concept of the beer-taste beverage and method for producing the beer-taste beverage of the present invention.

[0008] That is, although not limited to the following, the present invention relates to the following beer-taste beverages, etc. [1] A beer-flavored beverage having a purine content of 15 ppm or less and a peptide content of 35 to 50 kDa molecular weight of 1.5 to 50 ppm. [2] The beer-taste beverage according to [1] above, wherein the content of peptides having a molecular weight of 35 to 50 kDa is 7 to 50 ppm. [3] The beer-flavored beverage according to [1] or [2] above, having a xanthine content of 5 ppm or less. [4] The beer-taste beverage according to any one of [1] to [3] above, wherein the malt ratio is 25% by weight or more. [5] The beer-taste beverage according to any one of [1] to [4] above, wherein the malt ratio is 50% by weight or more. [6] The beer-taste beverage according to any one of [1] to [5] above, having a carbohydrate content of less than 0.5 g / 100 mL. [7] The beer-taste beverage according to any one of [1] to [6] above, having a total peptide content of 60 to 600 ppm. [8] A method for producing a beer-taste beverage, comprising the steps of adjusting the purine content to 15 ppm or less and adjusting the peptide content with a molecular weight of 35 to 50 kDa to 1.5 to 50 ppm. [Effects of the Invention]

[0009] The beer-taste beverage of the present invention has a reduced purine content and a specific range of peptide content with a molecular weight of 35 to 50 kDa, resulting in an excellent taste and a mild astringent flavor. Furthermore, the method for producing a beer-taste beverage of the present invention can provide a beer-taste beverage with a reduced purine content, an excellent taste, and a mild astringent flavor. DETAILED DESCRIPTION OF THE INVENTION

[0010] <<Beer-flavored beverages>> First, the beer-taste beverage of the present invention will be described. The beer-taste beverage of the present invention has a purine content of 15 ppm or less and a peptide content of 35 to 50 kDa of 1.5 to 50 ppm. Because the beer-taste beverage of the present invention has a reduced purine content and a peptide content of 35 to 50 kDa within a specific range, it has an excellent taste and little astringency.

[0011] As used herein, the term "beer-taste beverage" refers to an alcoholic or non-alcoholic carbonated beverage that has a beer-like flavor. In other words, unless otherwise specified, the term "beer-taste beverage" as used herein encompasses any carbonated beverage that has a beer flavor. Therefore, the term "beer-taste beverage" encompasses not only beer, a malt fermented beverage obtained by fermenting malt, hops, and water using yeast, but also fermented beer-taste beverages, as well as carbonated beverages to which beer flavorings containing esters, higher alcohols, lactones, etc. One embodiment of the beer-taste beverage of the present invention is beer. Furthermore, the "beer-taste beverage" may be a fermented beer-taste beverage that has undergone a fermentation process using yeast, or a non-fermented beer-taste beverage that has not undergone a fermentation process. Furthermore, the "beer-taste beverage" may be a beer-taste beverage containing distilled alcohol, such as spirits, whiskey, or shochu. Of these, spirit-containing beer-taste beverages are preferred.

[0012] As used herein, "taste" refers to the richness of flavor (depth and complexity) typical of a beer-flavored beverage. Furthermore, as used herein, "astringent taste" refers to a taste that gives a sensation of tightness in the mouth. Furthermore, as used herein, the presence and degree of "taste" and "astringent taste" can be evaluated by sensory evaluation by a specialist panel.

[0013] The beer-taste beverage of the present invention has a purine content of 15 ppm or less. As used herein, "purines" refers to eight compounds in total: four nucleosides, adenosine, guanosine, inosine, and xanthosine, and four purine bases, adenine, guanine, hypoxanthine, and xanthine. Furthermore, as used herein, "purine content" refers to the total content of adenosine, guanosine, inosine, xanthosine, adenine, guanine, hypoxanthine, and xanthine. In the beer-taste beverage of the present invention, the purine content is not particularly limited as long as it is 15 ppm or less, but from a health-conscious perspective, it is preferably 14 ppm or less, 13 ppm or less, 12 ppm or less, 11 ppm or less, 10 ppm or less, 9 ppm or less, 8 ppm or less, 7 ppm or less, 6 ppm or less, 5 ppm, 4 ppm or less, 3 ppm or less, 2 ppm or less, 1.5 ppm or less, or 1 ppm or less. The lower limit of the purine content is not particularly limited, but can be, for example, 0 ppm, 0.01 ppm, or 0.1 ppm. In this specification, the purine content in a beer-taste beverage refers to the value measured using a liquid chromatography-tandem mass spectrometer (LC-MS / MS) ("Guide to Microanalysis of Purines in Alcoholic Beverages," Japan Food Research Laboratories, Internet (https: / / www.jfrl.or.jp / storage / file / news_vol4_no23.pdf), searched March 2024). The beer-taste beverage of the present invention may be a low-purine beverage, a purine-free beverage, or a zero-purine beverage.

[0014] Among purines, nucleosides such as adenosine, guanosine, inosine, and xanthosine are known to be hydrolyzed by nucleosidase to form purine bases such as adenine, guanine, hypoxanthine, and xanthine, respectively, resulting in a reduction in their content. Among the purine bases, adenine, guanine, and hypoxanthine are known to be assimilated as nitrogen sources by common brewer's yeast (e.g., Saccharomyces) during the beer brewing process, resulting in a reduction in their content. On the other hand, common brewer's yeast cannot assimilate xanthine. Therefore, among purines (purine bases), only xanthine cannot be reduced in content by assimilation by common brewer's yeast. Therefore, in producing a beer-flavored beverage with a reduced purine content, a method that can reduce the xanthine content and thereby the purine content is extremely useful, apart from methods that may significantly impair the taste of the beer-flavored beverage, such as adsorption removal using adsorbents such as activated carbon or dilution. Prior to inventing the beer-taste beverage of the present invention, the inventors discovered that by carrying out a step of decomposing xanthine during the production of a beer-taste beverage, it is possible to reduce the xanthine content in the beer-taste beverage, and thereby the purine content. Details of the purine reduction method discovered by the inventors are described below.

[0015] In the beer-taste beverage of the present invention, the xanthine content is not particularly limited, but from a health-conscious perspective, it is preferably 5 ppm or less, 4 ppm or less, 3 ppm or less, 2 ppm or less, 1.5 ppm or less, or 1 ppm or less. The lower limit of the xanthine content is also not particularly limited, but can be, for example, 0 ppm, 0.01 ppm, or 0.1 ppm. In this specification, the xanthine content in beer-taste beverages refers to the value measured using a liquid chromatography-tandem mass spectrometer (LC-MS / MS) detection method ("Guide to Microanalysis of Purines in Alcoholic Beverages," Japan Food Research Laboratories, Internet (https: / / www.jfrl.or.jp / storage / file / news_vol4_no23.pdf), searched March 2024).

[0016] In this specification, "ppm" means "ppm by weight", and -4 Unless otherwise specified, "mg / 100mL" represents 10 -3 It is expressed in weight percent, i.e., 1 ppm = 1 mg / L = 0.1 mg / 100 mL = 0.0001 weight percent.

[0017] The beer-taste beverage of the present invention contains 1.5 to 50 ppm of peptides with a molecular weight of 35 to 50 kDa. Because the beer-taste beverage of the present invention contains peptides with a molecular weight of 35 to 50 kDa within the above range, the beer-taste beverage has a reduced purine content, yet has an excellent taste and little astringency. Peptides with a molecular weight of 35 to 50 kDa are derived from malt, etc. Peptides with a molecular weight of 35 to 50 kDa are peptides found in the 35 to 50 kDa molecular weight range when the raw material liquid of the beer-taste beverage is subjected to ultrafiltration using a 30 kDa cutoff membrane, followed by SDS-PAGE electrophoresis. Peptides of approximately 40 kDa are preferred, and peptides of approximately 40 kDa are also referred to as 40 kDa peptides in this specification. In the beer-taste beverage of the present invention, the content of the 40 kDa peptide is not particularly limited as long as it is 1.5 to 50 ppm, but may be, for example, 2 ppm or more, 2.5 ppm or more, 3 ppm or more, 3.5 ppm or more, 4 ppm or more, 4.5 ppm or more, 5 ppm or more, 5.5 ppm or more, 6 ppm or more, 7 ppm or more, 10 ppm or more, 15 ppm or more, 20 ppm or more, or 25 ppm or more, or may be 49 ppm or less, 48 ​​ppm or less, 47 ppm or less, 46 ppm or less, 45 ppm or less, or 40 ppm or less. In this specification, the content of the 40 kDa peptide refers to the value measured by the Bradford method.

[0018] The malt ratio of the beer-taste beverage of the present invention is not particularly limited, and may be 25% by mass or more, 30% by weight or more, 35% by weight or more, 40% by weight or more, 45% by weight or more, 50% by weight or more, 55% by weight or more, 60% by weight or more, 65% by weight or more, 66% by weight or more, more than 66% by weight, 67% by weight or more, 70% by weight or more, 75% by weight or more, 80% by weight or more, 85% by weight or more, 90% by weight or more, 95% by weight or more, or 100% by weight, or may be 100% by weight or less, less than 100% by weight, 98% by weight or less, 95% by weight or less, 90% by weight or less, 87% by weight or less, 85% by weight or less, 82% by weight or less, 80% by weight or less, 78% by weight or less, 76% by weight or less, 74% by weight or less, or less than 66% by weight. In this specification, "malt ratio" means a value calculated in accordance with the Liquor Tax Act and the Interpretation Notice of Liquor Administration-related Laws and Regulations, etc., which came into effect on April 1, 2018.

[0019] In this specification, "malt" refers to germinated seeds of wheat, such as barley, wheat, rye, oats, oats, adlay, and oats, which have been dried and have their roots removed, and may be of any origin or variety. Barley malt is one of the malts most commonly used as an ingredient in Japanese beer-flavored beverages. There are various types of barley, such as two-row barley and six-row barley, and either can be used. In addition to regular malt, colored malt can also be used. When using colored malt, different types of colored malt can be used in combination as appropriate, or a single type of colored malt can be used. In the beer-taste beverage of the present invention, the malt is preferably barley malt.

[0020] The beer-taste beverage of the present invention may also be one that uses grains other than malt, protein, sugar solution, etc. as ingredients other than malt. Examples of grains other than malt include barley (barley, wheat, rye, oats, oats, pearl barley, oats, etc.) that does not fall under the category of malt, rice (white rice, brown rice, etc.), corn (corn grits, etc.), sorghum, potatoes, beans (soybeans, peas, etc.), buckwheat, sorghum, millet, barnyard millet, and starch obtained therefrom, as well as extracts thereof. Among these, those using corn (corn grits, etc.) are preferred. Examples of proteins include soybean protein, pea protein, and their decomposition products.

[0021] The beer-taste beverage of the present invention may be an alcohol-containing beer-taste beverage with an alcohol content of 1% (v / v) or more, or a non-alcoholic beer-taste beverage with an alcohol content of less than 1% (v / v).The beer-taste beverage of the present invention is preferably an alcohol-containing beer-taste beverage.

[0022] As used herein, "alcohol-containing beer-flavored beverage" refers to a beer-flavored beverage with an alcohol content of 1% (v / v) or more. The source of the alcohol contained in an alcohol-containing beer-flavored beverage is not limited to fermented or non-fermented.

[0023] As used herein, the term "non-alcoholic beer-flavored beverage" refers to a beer-flavored beverage with an alcohol content of less than 1% (v / v), and includes any carbonated beverage that has a beer flavor and an alcohol content of less than 1% (v / v). "Non-alcoholic beer-flavored beverages" are not limited to fermented beverages produced by adding yeast to wort or a pre-fermentation liquid containing the components necessary for fermentation and fermenting the resulting mixture. They also include fermented and non-fermented carbonated beverages to which beer flavorings (flavorings that evoke a beer-like aroma) have been added, including esters, higher alcohols, and lactones, such as isoamyl acetate, ethyl acetate, n-propanol, isobutanol, acetaldehyde, ethyl caproate, ethyl caprylate, isoamyl propionate, linalool, geraniol, citral, 4-vinylguaiacol (4-VG), 4-methyl-3-pentenoic acid, 2-methyl-2-pentenoic acid, 1,4-cineole, 1,8-cineole, 2,3-diethyl-5-methylpyrazine, γ-decanolactone, γ-undecalactone, ethyl hexanoate, ethyl 2-methylbutyrate, ethyl n-butyrate, and myrcene. A "non-alcoholic beer-taste beverage" may be a fermented beverage obtained by fermenting using yeast (top-fermenting yeast and / or bottom-fermenting yeast) in the production process, followed by removal of the alcohol produced during the fermentation process, resulting in an alcohol content of less than 1 (v / v%). It may also be a fermented beverage obtained by terminating fermentation so that the alcohol content is less than 1 (v / v%). It may also be a fermented beverage diluted with water or other suitable ingredients so that the alcohol content is less than 1 (v / v%). When fermentation is terminated, it is preferable to terminate the fermentation so that the levels of off-flavors such as hydrogen sulfide, diacetyl, 2,3-pentanedione, and acetaldehyde are below a threshold, but this is not necessarily required. The concentration of off-flavors such as hydrogen sulfide, diacetyl, 2,3-pentanedione, and acetaldehyde is not limited as long as they blend with the flavor and aroma of the beer-taste beverage to create a favorable flavor. A "non-alcoholic beer-taste beverage" may also be a non-fermented beverage prepared without a fermentation process.

[0024] In this specification, the alcohol content of a non-alcoholic beer-taste beverage is less than 1 (v / v)%, and may be 0.8 (v / v)% or less, 0.6 (v / v)% or less, 0.5 (v / v)% or less, 0.4 (v / v)% or less, 0.2 (v / v)% or less, 0.1 (v / v)% or less, 0.05 (v / v)% or less, 0.01 (v / v)% or less, or 0.005 (v / v)% or less. Types of non-alcoholic beer-taste beverages include, for example, non-alcoholic beer-taste beverages and beer-taste soft drinks. In this specification, "alcohol content (alcohol content)" means the content of ethanol (v / v%) and does not include the content of aliphatic alcohols other than ethanol. In this specification, "volume %" and "v / v %" have the same meaning.

[0025] Yeast may be used to produce alcoholic and non-alcoholic beer-flavored beverages to produce fermented beer-flavored beverages. The fermented beer-flavored beverage may be a top-fermented beer-flavored beverage (ale beer-flavored beverage) brewed through a fermentation process using top-fermenting yeast (e.g., Saccharomyces), or a bottom-fermented beer-flavored beverage (lager beer-flavored beverage, pilsner beer-flavored beverage) brewed through a fermentation process using bottom-fermenting yeast (e.g., Saccharomyces), or a blend thereof. Fermentation may be performed using alcohol-producing yeast (e.g., Saccharomyces) or wild yeast (e.g., Brettanomyces), or may also use non-alcoholic yeast (e.g., Saccharomyces), wild yeast (e.g., Brettanomyces), or bacteria that perform lactic acid fermentation or gluconic acid fermentation.

[0026] The alcohol content is expressed as a percentage ((v / v)%) on a volume / volume basis. The alcohol content of a beverage can be measured by any known method, for example, according to "8.3 Alcohol" in the "Revised BCOJ Beer Analysis Methods, 2013, Supplement and Revision (edited by the International Technical Committee (Analysis Committee) of the Brewers Association of Japan, published by the Brewery Society of Japan, a public interest incorporated foundation)."

[0027] The alcohol content can be adjusted by adding diluted water or carbonated water, the type of raw materials (malt, corn grits, sugar solution, etc.), the amount of raw materials, the type of enzyme, the amount of enzyme added, the timing of adding the enzyme, the saccharification time in the brewing tank, the proteolysis time in the brewing tank, the pH in the brewing tank, the pH during the brewing process (the wort production process from the addition of malt to before the addition of yeast), the amount of acid added when adjusting the pH, the timing of pH adjustment (during brewing, during fermentation, at the end of fermentation, before beer filtration, after beer filtration, etc.), and the temperatures during wort preparation (including during saccharification). The temperature setting and holding time of the region, the original extract concentration of the pre-fermentation liquid, the original extract concentration during the fermentation process, the fermentation conditions (oxygen concentration, aeration conditions, yeast variety, amount of yeast added, number of yeast grown, timing of yeast removal, fermentation temperature, fermentation time, pressure setting, carbon dioxide concentration, etc.), addition of ethanol, addition of ethanol composition (raw material alcohol, spirits (distilled liquor) such as shochu, awamori, whiskey, brandy, vodka, rum, tequila, gin, etc., brewer's alcohol, etc.), etc. can be appropriately set.

[0028] When the beer-taste beverage of the present invention is an alcohol-containing beer-taste beverage, the alcohol content is not particularly limited as long as it is 1% by volume or more, and examples thereof include 1.5% by volume or more, 2.0% by volume or more, 2.5% by volume or more, 3.0% by volume or more, 3.5% by volume or more, 3.6% by volume or more, 3.7% by volume or more, 3.8% by volume or more, 3.9% by volume or more, 4.0% by volume or more, 4.1% by volume or more, 4.2% by volume or more, 4.3% by volume or more, 4.4% by volume or more, 4.5% by volume or more, 4.6% by volume or more, 4.7% by volume or more, 4.8% by volume or more, It is preferably 4.9% by volume or more, 5.0% by volume or more, 5.1% by volume or more, 5.2% by volume or more, 5.3% by volume or more, 5.4% by volume or more, 5.5% by volume or more, 5.6% by volume or more, 5.7% by volume or more, 5.8% by volume or more, 5.9% by volume or more, 6.0% by volume or more, 6.1% by volume or more, 6.2% by volume or more, 6.3% by volume or more, 6.4% by volume or more, 6.5% by volume or more, 7.0% by volume or more, 7.5% by volume or more, 8.0% by volume or more, 8.5% by volume or more, 9.0% by volume or more, 9.5% by volume or more, or 10.0% by volume or more. Furthermore, it is preferably 20.0 vol% or less, 19.5 vol% or less, 19.0 vol% or less, 18.5 vol% or less, 18.0 vol% or less, 17.5 vol% or less, 17.0 vol% or less, 16.5 vol% or less, 16.0 vol% or less, 15.5 vol% or less, 15.0 vol% or less, 14.5 vol% or less, 14.0 vol% or less, 13.5 vol% or less, 13.0 vol% or less, 12.5 vol% or less, 12.0 vol% or less, 11.5 vol% or less, 11.0 vol% or less, 10.5 vol% or less, or 10 vol% or less.

[0029] The manufacturing process for a typical beer-flavored beverage is shown below. First, a mixture containing malt and other barley, as well as other grains, starch, sugars, bittering agents, or coloring agents, and water, is gelatinized and saccharified, optionally with the addition of enzymes such as amylase. The mixture is then filtered to produce a saccharified liquid. Hops and bittering agents are added as needed, and the resulting saccharified liquid is boiled, with solids such as coagulated proteins removed in a clarifying tank. As an alternative to this saccharified liquid, hops can be added to malt extract and warm water, and then boiled. Hops can be added at any stage, from the beginning to the end of the boiling process. Known conditions can be used for the saccharification, boiling, solids removal, fermentation, and storage processes. The resulting fermented liquid is filtered, and carbon dioxide gas is added to the filtrate. The saccharified liquid is then filled into containers and sterilized to produce the desired beer-flavored beverage.

[0030] The beer-taste beverage of the present invention may or may not use hops as an ingredient. Hops may be in the form of pelleted hops, powdered hops, hop extract, etc. In addition, processed hop products such as isomerized hops and reduced hops may also be used. Furthermore, when hops are used as a raw material, the amount of hops used is not particularly limited, but is typically about 0.0001 to 1% by weight based on the total amount of the beer-flavored beverage.

[0031] The beer-taste beverage of the present invention may contain various additives as needed, provided that the effects of the present invention are not impaired. Examples of such additives include sweeteners (including high-intensity sweeteners), bittering agents or bitterness-imparting agents, flavorings, coloring agents, foam-forming agents, fermentation promoters, protein-based substances such as peptide-containing substances, dietary fiber, seasonings such as amino acids, antioxidants, and other additives. These may be used alone or in combination of two or more.

[0032] Examples of sweeteners include commercially available saccharified solutions obtained by hydrolyzing grain-derived starch with acid or enzymes, sugars such as sucrose and commercially available starch syrup, trisaccharides or higher, sugar alcohols, isomerized sugar, and natural sweeteners such as stevia, and artificial sweeteners. These sugars may be in the form of liquids such as solutions or solids such as powders. Furthermore, there are no particular limitations on the type of grain from which the starch is derived, the starch purification method, or the treatment conditions for enzymatic or acidic hydrolysis. For example, sugars with a higher maltose content may be used by appropriately setting the conditions for enzymatic or acidic hydrolysis. Other examples include sucrose, fructose, glucose, maltose, trehalose, maltotriose, maltotetraose, isomaltose, isomaltotriose, isomaltotetraose, and solutions (sugar solutions) of these. Examples of artificial sweeteners include aspartame, acesulfame potassium (acesulfame K), sucralose, and neotame. These sweeteners may be used alone or in combination of two or more.

[0033] The bittering agent or bitterness imparting agent is not particularly limited, and includes, in addition to hops, those used as bittering agents in ordinary beer and happoshu, such as rosemary, lychee, anise, juniper nut, sage, cinnamon, Ganoderma lucidum, bay leaf, mulberry, citrus extract, bitter persimmon extract, coffee extract, tea extract, bitter gourd extract, lotus germ extract, aloe arborescens extract, rosemary extract, lychee extract, laurel extract, sage extract, caraway extract, naringin, wormwood and wormwood extract, absinthin, alginic acid, etc. These bittering agents or bitterness imparting agents may be used alone or in combination of two or more.

[0034] The flavoring agent is not particularly limited, and general beer flavoring agents can be used. Beer flavoring agents are used to impart a beer-like flavor and include brewing components generated by fermentation. Examples of beer flavorings include isoamyl acetate, ethyl acetate, n-propanol, isobutanol, acetaldehyde, ethyl caproate, ethyl caprylate, isoamyl propionate, linalool, geraniol, citral, 4-vinylguaiacol (4-VG), 4-methyl-3-pentenoic acid, 2-methyl-2-pentenoic acid, 1,4-cineole, 1,8-cineole, 2,3-diethyl-5-methylpyrazine, γ-decanolactone, and γ-undecalactone. , Ethyl hexanoate, Ethyl 2-methylbutyrate, Ethyl n-butyrate, Myrcene, Citral, Limonene, Maltol, Ethyl maltol, Phenylacetic acid, Furaneol, Furfural, Methional, 3-methyl-2-butene-1-thiol, 3-methyl-2-butanethiol, Diacetyl, Ferulic acid, Geranic acid, Geranyl acetate, Ethyl butyrate, Octanoic acid, Decanoic acid, 9-decenoic acid, Nonanoic acid, Tetradecanoic acid, Propanoic acid, 2-methylpropanoic acid, γ-butyro Lactone, 2-aminoacetophenone, ethyl 3-phenylpropionate, 2-ethyl-4-hydroxy-5-methyl-3(2H)-furanone, dimethyl sulfone, 3-methylcyclopentane-1,2-dione, 2-methylbutanal, 3-methylbutanal, 2-methyltetrahydrofuran-3-one, 2-acetylfuran, 2-methyltetrahydrofuran-3-one, hexanal, hexanol, cis-3-hexenal, 1-octen-3-ol, β-eutectic alcohol Desmol, 4-mercapto-4-methylpentan-2-one, β-caryophyllene, β-myrcene, furfuryl alcohol, 2-ethylpyrazine, 2,3-dimethylpyrazine, 2-methylbutyl acetate, isoamyl alcohol, 5-hydroxymethylfurfural, phenylacetaldehyde, 1-phenyl-3-buten-1-one, trans-2-hexenal, nonanal, phenethyl alcohol, nerol, citronellol, methyl p-toluate, 1,2,3,5-Tetramethylbenzene, triethyl citrate, tributyl citrate, diethyl tartrate, dibutyl malate, perillaldehyde, methylheptenone, lemon myrtle, cinnamaldehyde, 2-propanol, n-butanol, 2-butanol, activated amyl alcohol, 2-heptanol, 2-octanol, 5-methylfurfuryl alcohol, ethyl butyrate, ethyl isobutyrate, ethyl heptanoate, isobutyl acetate, activated amyl acetate, propionate Examples of suitable amines include ethyl octanoate, isoamyl octanoate, 2,3-butanediol, methionol, isovaleraldehyde, pentanal, octanal, 2-nonanal, trans-2-nonenal, methylfurfural, β-ionone, hydroxymethylfurfural, acetoin, 2,3-butanedione, 2,3-pentanedione, 2,3-hexanedione, guaiacol, α-terpineol, damascenone, nerolidol, humulene, and linalool oxide. These fragrances may be used alone or in combination of two or more.

[0035] Coloring agents are used to impart a beer-like color to beverages, and examples thereof include caramel color, lycopene color, elderberry color, cocoa color, safflower color, etc. These coloring agents may be used alone or in combination of two or more.

[0036] Foam-forming agents are used to form beer-like foam in beverages or to maintain the foam of beverages, and examples include plant-extracted saponin substances such as soybean saponin and quillaja saponin, plant proteins such as corn and soybean, peptide-containing substances such as collagen peptide, raw materials derived from milk, etc. These foam-forming agents may be used alone or in combination of two or more.

[0037] Fermentation promoters are used to promote fermentation by yeast, and examples thereof include bran components from rice, wheat, etc., vitamins, mineral supplements, etc. These fermentation promoters may be used alone or in combination of two or more.

[0038] Examples of dietary fibers include water-soluble dietary fibers. Examples of water-soluble dietary fibers include indigestible dextrin, polydextrose, guar gum hydrolysate, pectin, glucomannan, alginic acid, laminarin, fucoidin, and carrageenan. From the viewpoint of versatility such as stability and safety, indigestible dextrin or polydextrose is preferred. These dietary fibers may be used alone or in combination of two or more.

[0039] The antioxidant is not particularly limited, and those used as antioxidants in ordinary beer and happoshu can be used, such as ascorbic acid, erythorbic acid, catechin, etc. These antioxidants may be used alone or in combination of two or more.

[0040] Other additives include, but are not limited to, fruits (including dried or boiled fruits, or concentrated fruit juice); coriander or its seeds; pepper, cinnamon, cloves, Japanese pepper and other spices or their raw materials; chamomile, sage, basil, lemongrass and other herbs; sweet potato, pumpkin and other vegetables (including dried or boiled vegetables); buckwheat or sesame; honey and other sugar-containing substances, salt or miso; flowers or tea, coffee, cocoa or preparations thereof; oysters, kelp, wakame seaweed or bonito flakes, etc. When the beer-taste beverage of the present invention contains other additives, the content thereof is not particularly limited, but is typically 5% by weight or less based on the weight of the malt used as an ingredient in the beer-taste beverage.

[0041] In the production of beer-taste beverages, by keeping the malt ratio low and by carrying out steps such as fermentation (purine assimilation by microorganisms such as yeast, including xanthine assimilation by microorganisms capable of xanthine assimilation (step (A1) in the purine reduction method described below)) at any time from the brewing process to the filling process, xanthine degradation using xanthine oxidase (step (A2) in the purine reduction method described below, etc.), hydrolysis using nucleosidase (step (B) in the purine reduction method described below, etc.), adsorption and removal of purines using an adsorbent, and dilution, it is possible to obtain beer-taste beverages with a purine content reduced to 15 ppm or less. These steps may be carried out alone or in combination of two or more. In one embodiment of the present invention, the fermentation step (purine assimilation by microorganisms such as yeast) can be performed using, for example, a method similar to step (A1) (a step of causing a microorganism capable of assimilating xanthine to assimilate xanthine) in the purine reduction method described below. In one embodiment of the present invention, the xanthine degradation step using xanthine oxidase can be performed using, for example, a method similar to step (A2) (a step of decomposing xanthine using xanthine oxidase) in the purine reduction method described below. In addition, the hydrolysis step using nucleosidase can be performed using, for example, a method similar to step (B) (a step of adding nucleosidase) in the purine reduction method described below. Furthermore, the steps of fermentation (purine assimilation by microorganisms such as yeast), xanthine degradation using xanthine oxidase, hydrolysis using nucleosidase, adsorption / removal of purines using an adsorbent, and dilution are not particularly limited, and conventionally known methods and conditions can be used. The adsorbent used for adsorbing and removing purines is not particularly limited, and examples thereof include activated carbon, zeolite, activated clay, and the like. The beer-taste beverage of the present invention is preferably produced through the purine reduction method described below.

[0042] When the beer-taste beverage of the present invention is intended to be an alcohol-containing beer-taste beverage, the alcohol content of the final product can be adjusted by adding a raw material alcohol, etc. The raw material alcohol may be added at any step from the saccharification step to the filling step.

[0043] An aliphatic alcohol may be added to the beer-taste beverage of the present invention in order to impart a boozy flavor. There are no particular limitations on the aliphatic alcohol as long as it is a known alcohol, but an aliphatic alcohol having 4 to 5 carbon atoms is preferred. Examples of aliphatic alcohols having 4 carbon atoms include 2-methyl-1-propanol and 1-butanol. Examples of aliphatic alcohols having 5 carbon atoms include 3-methyl-1-butanol, 1-pentanol, and 2-pentanol. These may be used alone or in combination of two or more.

[0044] In the beer-taste beverage of the present invention, the content of the aliphatic alcohol having 4 to 5 carbon atoms is not particularly limited, but is preferably 0.0002 to 0.0007% by weight, and more preferably 0.0003 to 0.0006% by weight. In this specification, the content of the aliphatic alcohol having 4 to 5 carbon atoms can be measured using headspace gas chromatography.

[0045] The beer-taste beverage of the present invention may be a low-carbohydrate beverage or a reduced-carbohydrate beverage in line with the recent trend toward low-carbohydrate beverages. In the beer-taste beverage of the present invention, the carbohydrate content can be set according to the desired characteristics of the beverage, and may be, for example, 2.0 g / 100 mL or less, 1.9 g / 100 mL or less, 1.8 g / 100 mL or less, 1.7 g / 100 mL or less, 1.6 g / 100 mL or less, 1.5 g / 100 mL or less, less than 1.5 g / 100 mL, 1.4 g / 100 mL or less, 1.3 g / 100 mL or less, 1.2g / 100mL or less, 1.1g / 100mL or less, 1.0g / 100mL or less, 1.0g / 100mL or less, 0.95g / 100mL or less, 0.9g / 100mL or less, 0.85 g / 100mL or less, 0.8g / 100mL or less, 0.75g / 100mL or less, 0.7g / 100mL or less, 0.65g / 100mL or less, 0.6g / 100mL or less, 0.55g / It may be 100mL or less, 0.5g / 100mL or less, or less than 0.5g / 100mL, or it may be 0.1g / 100mL or more, 0.2g / 100mL or more, 0.3g / 100mL or more, 0.4g / 100mL or more, 0.5g / 100mL or more, more than 0.5g / 100mL, 0.55g / 100mL or more, 0.6g / 100mL or more, 0.65g / 100mL or more, or 0. It may be 7g / 100mL or more, 0.75g / 100mL or more, 0.8g / 100mL or more, 0.85g / 100mL or more, 0.9g / 100mL or more, 0.95g / 100mL or more, 1.0g / 100mL or more, 1.1g / 100mL or more, 1.2g / 100mL or more, 1.3g / 100mL or more, 1.4g / 100mL or more, or 1.5g / 100mL or more. From a health-conscious perspective, the beer-taste beverage of the present invention preferably has a carbohydrate content of less than 0.5 g / 100 mL.

[0046] In this specification, "carbohydrates" refers to carbohydrates based on the Food Nutrition Labeling Standards (Ministry of Health, Labour and Welfare Notification No. 176 of 2003, partially revised by Consumer Affairs Agency Notification No. 8 of September 27, 2013), and specifically refers to the carbohydrates remaining after excluding protein, lipids, dietary fiber, ash, alcohol and water from the food in question. The carbohydrate content of a food product can be calculated by subtracting the amounts of protein, fat, dietary fiber, ash, and water from the weight of the food product. The amounts of protein, lipid, dietary fiber, ash, and moisture can be measured by the methods specified in the Nutrition Labeling Standards. Specifically, the amount of protein can be measured by the nitrogen quantitative conversion method, the amount of lipid by the ether extraction method, the amount of dietary fiber by the Prosky method, the amount of ash by the direct ashing method, and the amount of moisture by the reduced pressure heat drying method.

[0047] In addition to the 40 kDa peptide described above, the beer-taste beverage of the present invention contains other peptides derived from malt, etc. The other peptides are peptides with molecular weights of less than 35 kDa or more than 50 kDa, and more specifically, refer to peptides other than the 40 kDa peptides found in the molecular weight range of 35 to 50 kDa when the raw material liquid of the beer-taste beverage is subjected to ultrafiltration using a 30 kDa cutoff membrane and then subjected to SDS-PAGE electrophoresis.In the beer-taste beverage of the present invention, the total peptide content, which is the combined content of the 40 kDa peptide and other peptides, is not particularly limited, and may be, for example, 60 ppm or more, 61 ppm or more, 62 ppm or more, 63 ppm or more, 64 ppm or more, 65 ppm or more, 66 ppm or more, 67 ppm or more, 68 ppm or more, 69 ppm or more, 70 ppm or more, 71 ppm or more, 72 ppm or more, 73 ppm or more, 74 ppm or more, or 75 ppm or more. , 76ppm or more, 77ppm or more, 78ppm or more, 79ppm or more, 80ppm or more, 81ppm or more, 82ppm or more, 83ppm or more, 84ppm or more, 85ppm or more, 86ppm or more, 87ppm or more, 88ppm or more, 89ppm or more, 90ppm or more, 91ppm or more, 92ppm or more, 93ppm or more, 94ppm or more, 95ppm or more, 96ppm or more, 97ppm or more, 98ppm or more, 99ppm or more, 100ppm m or more, 110 ppm or more, 120 ppm or more, 130 ppm or more, 140 ppm or more, 150 ppm or more, 160 ppm or more, 170 ppm or more, 180 ppm or more, 190 ppm or more, or 200 ppm or more, and may be 600 ppm or less, 590 ppm or less, 580 ppm or less, 570 ppm or less, 560 ppm or less, 550 ppm or less, 540 ppm or less, 530 ppm or less, 520 ppm or less, 510 ppm or less, 5 The concentration may be 00 ppm or less, 490 ppm or less, 480 ppm or less, 470 ppm or less, 460 ppm or less, 450 ppm or less, 440 ppm or less, 430 ppm or less, 420 ppm or less, 410 ppm or less, 400 ppm or less, 390 ppm or less, 380 ppm or less, 370 ppm or less, 360 ppm or less, 350 ppm or less, 340 ppm or less, 330 ppm or less, 320 ppm or less, 310 ppm or less, or 300 ppm or less. In this specification, the total peptide content refers to the value measured by the Lowry method.

[0048] In the beer-taste beverage of the present invention, the total polyphenol content is not particularly limited, but from the standpoint of taste, it may be, for example, 40 ppm or more, 50 ppm or more, 60 ppm or more, 70 ppm or more, 80 ppm or more, 90 ppm or more, 100 ppm or more, or 110 ppm or more, and may be 250 ppm or less, 220 ppm or less, 200 ppm or less, 180 ppm or less, 160 ppm or less, 150 ppm or less, 140 ppm or less, 130 ppm or less, or 120 ppm or less. In this specification, the total polyphenol content refers to the total content of polyphenols contained in the beer-taste beverage. Examples of polyphenols include, but are not limited to, xanthohumol, catechin, hesperidin, chlorogenic acid, resveratrol, quercetin, anthocyanogen, and tannin. The total polyphenol content in a beer-flavored beverage can be measured, for example, by the method described in Section 8.19 Total Polyphenols in the Revised BCOJ Beer Analysis Methods (published by the Brewery Association of Japan, edited by the International Technical Committee (Analysis Committee) of the Brewers Association of Japan, revised and expanded in 2013). The total polyphenol content can be adjusted by adding dilution water or carbonated water, the type of raw material (raw materials containing polyphenols such as malt, corn grits, sugar solution, etc.), the amount of raw material, the type of enzyme, the amount of enzyme added, the timing of enzyme addition, the polyphenol polymerization conditions in the mash tank (temperature, stirring speed, etc.), the aeration time in the mash tank (mash aeration, etc.), the pH in the mash tank, the pH during the mashing process (the wort production process from the addition of malt to before the addition of yeast), the time for filtering the wort, the set temperature and holding time for each temperature range when preparing the wort (including during saccharification), and the temperature of the pre-fermentation liquid. The original extract concentration, the original extract concentration during the fermentation process, fermentation conditions (oxygen concentration, aeration conditions, yeast variety, amount of yeast added, yeast growth rate, timing of yeast removal, fermentation temperature, fermentation time, pressure setting, carbon dioxide concentration, etc.), cooling timing, cooling temperature, cooling time, type of beer filtration (diatomaceous earth, membrane, sheet, cartridge, filter, etc.), activated carbon, stabilizers added during beer filtration (silica gel, PVPP (polyvinylpolypyrrolidone), bentonite, tannin, bentonite, etc.), etc. can be appropriately set. Furthermore, the total polyphenol content of the beer-taste beverage of the present invention can be controlled by adjusting the amount of raw materials with high polyphenol content, such as barley malt, malt husks, etc. Specifically, the total polyphenol content can be increased by increasing the amount of raw materials with high polyphenol content, such as malt. Generally, malt containing husks (grain husks) has a high nitrogen and polyphenol content, while soybeans, yeast extract, peas, corn, processed corn products (corn grits, corn protein, etc.), wheat, and wheat malt have a high nitrogen content but a low polyphenol content. Therefore, the total nitrogen and polyphenol content of a beer-flavored beverage can be increased or decreased by adjusting the blending ratio of the ingredients. Representative methods (1) to (4) for increasing or decreasing the total nitrogen and polyphenol content are listed below. (1) By increasing the amount of husk-containing malt used, the total nitrogen and polyphenol content of beer-flavored beverages can be increased. (2) By increasing or decreasing the amount of soybeans, yeast extract, etc. used, the total nitrogen content of the beer-flavored beverage can be increased or decreased while maintaining the total polyphenol content. (3) By increasing the amount of malt containing husk and decreasing the amount of soybeans, yeast extract, etc., the total polyphenol content can be increased while maintaining the total nitrogen content. (4) By reducing the amount of malt containing husk and increasing the amount of soybeans, yeast extract, etc. used, the total polyphenol content is reduced while maintaining the total nitrogen content.

[0049] In the beer-taste beverage of the present invention, the concentration of ammonia nitrogen is not particularly limited, but is preferably 380 μmol / L or less, more preferably 350 μmol / L or less, even more preferably 250 μmol / L or less, even more preferably 200 μmol / L or less, even more preferably 150 μmol / L or less, even more preferably 100 μmol / L or less, even more preferably 50 μmol / L or less, particularly preferably 25 μmol / L or less, and most preferably 10 μmol / L or less. The lower limit of the ammonia nitrogen concentration is also not particularly limited, but can be, for example, 0 μmol / L or 0.01 μmol / L. In this specification, ammonia nitrogen refers to nitrogen present in the form of ammonium ions in a beer-taste beverage. In this specification, the concentration of ammonia nitrogen refers to a value measured by high performance liquid chromatography (HPLC) using a high-speed amino acid analyzer L-8900 (manufactured by Hitachi High-Tech Corporation) under the conditions described in the Examples below. Methods for reducing the concentration of ammonia nitrogen in a beer-taste beverage include, for example, step (A1) of the purine reduction method described below, an ion exchange method using zeolite, and a reverse osmosis membrane method. However, the method of step (A1) of the purine reduction method described below is preferred because it can efficiently reduce the concentration of ammonia nitrogen.

[0050] The total nitrogen content of the beer-taste beverage of the present invention is not particularly limited and may be, for example, 15 mg / 100 mL or more, 20 mg / 100 mL or more, or 25 mg / 100 mL or more, or 100 mg / 100 mL or less, 95 mg / 100 mL or less, 90 mg / 100 mL or less, 85 mg / 100 mL or less, or 80 mg / 100 mL or less. As used herein, the total nitrogen content refers to the total amount of all nitrogen compounds, such as proteins, amino acids, and the aforementioned compounds containing ammonia nitrogen. The total nitrogen content of the beer-taste beverage of the present invention can be measured, for example, by the method described in "8.9 Total Nitrogen" in the Revised BCOJ Beer Analysis Methods (published by the Brewery Society of Japan, edited by the International Technical Committee of the Brewers Association of Japan (Analysis Committee) and expanded and revised in 2013). The total nitrogen amount can be adjusted by appropriately setting the beer filtration conditions, such as the addition of dilution water or carbonated water, the type of raw materials (malt, corn grits, sugar solution, etc.), the amount of raw materials, the type of enzyme, the amount of enzyme (including proteolytic enzymes, etc.) added, the temperature during the enzyme reaction, the timing of the enzyme addition, the proteolysis time in the mash tank, the pH in the mash tank, the temperature in the mash tank, the pH during the mashing process (the wort production process from the addition of malt to before the addition of yeast), the temperature during the mashing process, the temperature during wort filtration, the time of wort filtration, the pH during wort filtration, the amount of sparging water used during wort filtration, the set temperatures and holding times for each temperature range when preparing wort, the boiling time and pH in the boiling process, the original extract concentration of the pre-fermentation liquid, the original extract concentration in the fermentation process, and the fermentation conditions (oxygen concentration, aeration conditions, yeast variety, amount of yeast added, yeast growth rate, timing of yeast removal, fermentation temperature, fermentation time, pressure setting, carbon dioxide concentration, etc.).

[0051] In the beer-taste beverage of the present invention, the free amino nitrogen (FAN) content is not particularly limited and may be, for example, 0.5 mg / 100 mL or more, 1.0 mg / 100 mL or more, 1.5 mg / 100 mL or more, 2.0 mg / 100 mL or more, or 2.5 mg / 100 mL or more, or may be 15 mg / 100 mL or less, 14 mg / 100 mL or less, 13 mg / 100 mL or less, 12 mg / 100 mL or less, or 11 mg / 100 mL or less. The FAN content of the beer-taste beverage of the present invention can be measured, for example, by the method described in "8.18 Free amino nitrogen - ninhydrin colorimetric method" in the Revised BCOJ Beer Analysis Methods (published by the Brewery Society of Japan, a public interest incorporated foundation, edited by the International Technical Committee of the Brewers Association of Japan (Analysis Committee), revised and expanded in 2013). The FAN content can be adjusted by appropriately setting the following: adding dilution water or carbonated water, adding distilled alcohol such as spirits, adding brewed alcohol such as brewer's alcohol, type of raw materials (malt, corn grits, sugar solution, etc.), amount of raw materials, type of enzyme, amount of enzyme (including proteolytic enzymes, etc.) added, temperature during enzyme reaction, timing of enzyme addition, proteolysis time in the brewing tank, pH in the brewing tank, temperature in the brewing tank, pH during the brewing process (the wort production process from adding malt to before adding yeast), temperature during the brewing process, time for wort filtration, set temperatures and holding times for each temperature range when preparing wort, boiling time and pH in the boiling process, original extract concentration in the pre-fermentation liquid, original extract concentration in the fermentation process, fermentation conditions (oxygen concentration, aeration conditions, yeast variety, amount of yeast added, yeast growth rate, timing of yeast removal, fermentation temperature, fermentation time, pressure setting, carbon dioxide concentration, etc.).

[0052] The beer-taste beverage of the present invention may be colorless and transparent, or may be colored. In this specification, the "color" of a beverage can be measured by the measurement method described in "8.8 Color" of the Revised BCOJ Beer Analysis Methods (published by the Brewery Association of Japan, edited by the International Technical Committee (Analysis Committee) of the Brewers Association of Japan, revised and expanded in 2013). The "color" of a beverage is specified by the color unit (EBC unit) established by the European Brewery Convention. The smaller the value, the lighter the beverage, and conversely, the larger the value, the darker the beverage. The color may be 0EBC, 1EBC or more, 5EBC or more, 10EBC or more, 15EBC or more, 20EBC or more, 30EBC or more, or 40EBC or more. It may also be 200EBC or less, 150EBC or less, 100EBC or less, 50EBC or less, 30EBC or less, 20EBC or less, 15EBC or less, or 10EBC or less. The color can be adjusted by appropriately setting the addition of dilution water or carbonated water, the type of raw materials (malt, corn grits, sugar solution, etc.), the amount of raw materials, the mashing temperature, the pH during mashing, the mashing time, the wort filtration time, the pH during wort filtration, the boiling time, the boiling temperature, the amount of color components such as caramel color, the type of beer filtration (diatomaceous earth filtration, various membrane filtration, etc.), the amount of beer filtration, etc. The color of the beer-taste beverage of one embodiment of the present invention can be controlled by, for example, appropriately adjusting the type of malt used, the blending ratio when two or more types of malt are used, the ratio of malt to other ingredients, the boiling conditions when preparing the pre-fermentation solution, etc. More specifically, for example, the color of the beer-taste beverage can be increased by increasing the blending ratio of dark-colored malt, increasing the boiling temperature, extending the boiling time, or performing decoction when preparing the saccharified solution. The color can also be increased by increasing the concentration of raw wort extract or the proportion of malt. The color can also be adjusted by controlling the amount of food additives such as caramel color or colored sugar solution.

[0053] The beer-taste beverage of one embodiment of the present invention may be a packaged beverage. Packages of any shape and material may be used for packaged beverages. Examples of containers include glass bottles, cans, barrels, and plastic bottles. Cans, glass bottles, and plastic bottles are preferred from the viewpoint of portability.

[0054] Next, the above-mentioned method for reducing purines discovered by the present inventors will be described in detail. The purine reduction method discovered by the present inventors (hereinafter also referred to simply as the purine reduction method) includes a step (A) of decomposing xanthine. By carrying out the purine reduction method at any time during the production of a beer-taste beverage, from the brewing step to the filling step, the purine (xanthine) content in the beer-taste beverage can be reduced.

[0055] <Process (A)> Step (A) is a step of decomposing xanthine. In this specification, decomposing xanthine refers to decomposing xanthine at least to uric acid, and includes assimilation of xanthine. Step (A) of decomposing xanthine is not particularly limited as long as it can decompose xanthine at least to uric acid. Specific examples of step (A) of decomposing xanthine include step (A1) of allowing a microorganism capable of assimilating xanthine to assimilate xanthine, and step (A2) of decomposing xanthine using xanthine oxidase. In the method for reducing purines, step (A) may be performed by either step (A1) or step (A2) alone, or by combining step (A1) and step (A2). Furthermore, when step (A1) and step (A2) are performed in combination, the order in which they are performed is not particularly limited; step (A1) and step (A2) may be performed simultaneously, step (A1) may be followed by step (A2), or step (A1) may be performed after step (A2). Step (A1) and step (A2) will be described in order below.

[0056] Step (A1): A step of allowing a microorganism capable of assimilating xanthine to assimilate xanthine In the method for reducing purines, step (A) can include a step (A1) of allowing a microorganism capable of assimilating xanthine to assimilate xanthine. The microorganism capable of xanthine assimilation is not particularly limited as long as it is capable of assimilating xanthine, and may be a eukaryote including fungi such as yeast, or a prokaryote such as bacteria or archaea, but is preferably a yeast capable of xanthine assimilation. As the microorganism capable of xanthine assimilation, one type may be used alone, or two or more types may be used in combination.

[0057] The microorganism capable of assimilating xanthine is preferably a microorganism having the XAN2 gene, and more preferably a yeast having the XAN2 gene. The XAN2 gene is a gene encoding xanthine oxidase. Xanthine oxidase is an enzyme that catalyzes the oxidation reaction of hypoxanthine to xanthine and the oxidation reaction of xanthine to uric acid. A microorganism (yeast) having the XAN2 gene can assimilate xanthine through the action of xanthine oxidase.

[0058] Examples of yeasts capable of assimilating xanthine include those of the genus Torulaspora, Lachancea, Zygosaccharomyces, Kluyveromyces, Hanseniaspora, Barnettozyma, Cyberlindnera, Wickerhamomyces, Ascoidea, Saccharomycopsis, Babjeviella, Candida / Lodderomyces, Debaryomyces, Diutina, Hyphopichia, Meyerozyma, Millerozyma, Sche Examples of yeasts that belong to the genera Fersomyces, Spathaspora, Suhomyces, Yamadazyma, Clavispora, Metschnikowia, Citeromyces, Komagataella, Kuraishia, Ambrosiozyma, Brettanomyces, Ogataea, Pichia, Saturnispora, Nakazawaea, Pachysolen, Peterozyma, Nadsonia, Galactomyces, Saprochaete, Yarrowia, Sugiyamaella, Tortispora, and Lipomyces. All yeasts that belong to these genera have the XAN2 gene. Examples of yeasts belonging to the genus Torulaspora include Torulaspora delbrueckii and Torulaspora pretoriensis. Examples of yeasts belonging to the genus Lachancea include Lachancea fermentati and Lachancea thermotolerans. Examples of yeasts belonging to the genus Zygosaccharomyces include Zygosaccharomyces rouxii. Examples of yeasts belonging to the genus Kluyveromyces include Kluyveromyces marxianus. In the method for reducing purines, the yeast capable of xanthine assimilation is preferably at least one species selected from the group consisting of yeasts belonging to the genus Torulaspora, yeasts belonging to the genus Lachancea, yeasts belonging to the genus Zygosaccharomyces, and yeasts belonging to the genus Kluyveromyces, and examples thereof include Torulaspora delbrueckii, Torulaspora pretoriensis, Lachancea fermentati, Lachancea thermotolerans, Zygosaccharomyces rouxii, and Kluyveromyces marxianus. marxianus, more preferably at least one selected from the group consisting of Torulaspora delbrueckii, Lachancea fermentati, Zygosaccharomyces rouxii, and Kluyveromyces marxianus, and particularly preferably Torulaspora delbrueckii.

[0059] The microorganisms capable of xanthine assimilation may be commercially available products or may be those isolated and purified from nature. Alternatively, these microorganisms capable of xanthine assimilation may be grown in advance by pre-culture, and the resulting pre-culture solution may be used in step (A1). Furthermore, when the yeast is dried, it may be reconstituted with water by a conventionally known method before use in step (A1).

[0060] When a microorganism capable of xanthine assimilation is pre-cultured, the culture conditions are not particularly limited as long as the microorganism can grow, and can be appropriately set depending on the type of microorganism, etc. The culture temperature may be, for example, 10 to 35°C, and preferably 20 to 30°C. The culture time may be, for example, 5 hours to 5 days, and preferably 16 hours to 3 days.

[0061] In step (A1), examples of methods for allowing a microorganism capable of xanthine assimilation to assimilate xanthine include a method in which a microorganism capable of xanthine assimilation is added to wort or a wort fermentation liquid, and then fermentation is carried out by the microorganism in the wort or the wort fermentation liquid.

[0062] In step (A1), when a microorganism capable of xanthine assimilation is added to wort or a wort fermentation liquor, the wort or wort fermentation liquor preferably contains xanthine. The xanthine content in the wort or wort fermentation liquor is not particularly limited, but may be, for example, 0.1 to 50 ppm, and preferably 5 to 30 ppm.

[0063] In the step (A1), when a microorganism capable of xanthine assimilation is added to the wort or the wort fermentation broth, the amount of the microorganism capable of xanthine assimilation added is not particularly limited. For example, 1 to 200 × 10 microorganisms can be added to the wort or the wort fermentation broth. 6 cells / mL, and may be 10 to 100 × 10 6 cells / mL is preferred.

[0064] Fermentation conditions for allowing a microorganism capable of xanthine assimilation to assimilate xanthine are not particularly limited as long as the microorganism can assimilate xanthine, and can be appropriately set depending on the type and amount of the microorganism capable of xanthine assimilation, the xanthine content in the wort or fermented wort broth, etc. The fermentation temperature may be, for example, 5 to 35°C, and preferably 10 to 30°C. The fermentation time may be, for example, 5 hours to 10 days, and preferably 16 hours to 5 days.

[0065] In the purine reduction method, step (A1) may be performed at any timing during the production process of a beer-taste beverage, from the saccharification step to the filling step. For example, a microorganism capable of xanthine assimilation may be added to wort simultaneously with a conventional brewer's yeast and fermented (primary fermentation), or a microorganism capable of xanthine assimilation may be added to a wort fermentation liquid obtained by fermentation using a conventional brewer's yeast and further fermented (secondary fermentation).

[0066] Step (A2): Decomposing xanthine using xanthine oxidase In the method for reducing purines, step (A) can include step (A2) of decomposing xanthine using xanthine oxidase. The xanthine oxidase may be a commercially available product, or may be one isolated and purified from nature. Furthermore, xanthine oxidase derived from the aforementioned microorganisms capable of xanthine assimilation (microorganisms having the XAN2 gene) may also be used. Examples of xanthine oxidase derived from microorganisms capable of xanthine assimilation (microorganisms having the XAN2 gene) include those obtained by disrupting microorganisms capable of xanthine assimilation (microorganisms having the XAN2 gene) or their precultures using a conventionally known cell disruption method (crude enzyme solution), those obtained by further extracting the contents of the microorganisms capable of xanthine assimilation (microorganisms having the XAN2 gene) from the crude enzyme solution (cell extract), and those obtained by removing cell residues from the crude enzyme solution or the cell extract. When step (A2) is carried out as step (A) in the purine reduction method, the xanthine oxidase is preferably derived from a microorganism capable of assimilating xanthine (a microorganism having the XAN2 gene).

[0067] In the step (A2), examples of the method for decomposing xanthine using xanthine oxidase include a method in which xanthine oxidase is added to wort or a fermented wort broth. The conditions in step (A2), such as temperature, pH, and reaction time, are not particularly limited as long as the activity of xanthine oxidase is maintained and the enzymatic reaction (decomposition of xanthine) proceeds. For example, the temperature may be 5 to 35°C, preferably 10 to 30°C, and the reaction time may be 5 hours to 10 days, preferably 16 hours to 5 days.

[0068] When xanthine oxidase is added to the wort or the wort fermentation liquor in step (A2), the wort or the wort fermentation liquor preferably contains xanthine. The content of xanthine in the wort or the wort fermentation liquor is not particularly limited, but may be, for example, 0.1 to 50 ppm, and preferably 5 to 30 ppm.

[0069] When xanthine oxidase is added to the wort or wort fermentation liquor in step (A2), the amount added is not particularly limited, but is preferably an amount such that the activity of xanthine oxidase is, for example, 0.01 U or more, 0.05 U or more, 0.1 U or more, 0.3 U or more, 0.5 U or more, 0.7 U or more, 1.0 U or more, 1.5 U or more, 2.0 U or more, 2.5 U or more, 3.0 U or more, 3.5 U or more, 4.0 U or more, 5.0 U or more, 5.5 U or more, 6.0 U or more, 7.0 U or more, 8.0 U or more, 9.0 U or more, or 10.0 U or more, or 10,000 U or less, 9,000 U or less, 8,000 U or less, 7,000 U or less, or 6,000 U or less. In this specification, the activity value of xanthine oxidase means the relative enzyme amount when the amount of enzyme that produces 1 μmol of uric acid per minute under the following conditions is defined as 1 unit (U). Mix 2.9 ml of 50 mM Tris-HCl buffer and 0.1 ml of 10 mM xanthine aqueous solution and preheat at 37°C. Then, add 0.01 ml of the solution to be measured and gently mix. After that, measure the change in absorbance at 293 nm per minute using water as a control with a spectrophotometer controlled at 37°C. From this value, the amount of enzyme that produces 1 μmol of uric acid per minute is defined as 1 unit (U), and this is taken as the activity value of xanthine oxidase.

[0070] In the purine reduction method, step (A2) may be performed at any timing during the production process of a beer-taste beverage, from the saccharification step to the filling step. For example, xanthine oxidase may be added to wort simultaneously with conventional brewer's yeast to decompose xanthine, or xanthine oxidase may be added to a wort fermentation liquid obtained by fermentation using conventional brewer's yeast to decompose xanthine.

[0071] In the method for reducing purine content, it is preferable to add a microorganism capable of assimilating xanthine or xanthine oxidase to the wort or fermented wort broth in step (A).

[0072] In the purine reduction method, by carrying out step (A1) or step (A2) as step (A), it is possible to decompose xanthines and reduce the purine (xanthine) content in the beer-taste beverage. Furthermore, when step (A1) or step (A2) is performed as step (A), the content of components that contribute to the taste of the beer-taste beverage (the content of the 40 kDa peptide and the total peptide content) does not decrease significantly before or after step (A1) or step (A2), and therefore the content of purines (xanthines) can be reduced without significantly impairing the taste of the beer-taste beverage. In the method for reducing purine content, the step (A) of decomposing xanthine is preferably a step (A1) of allowing a microorganism capable of assimilating xanthine to assimilate xanthine.

[0073] <Process (B)> The purine reduction method preferably further comprises, in addition to the above-described step (A), step (B) of adding nucleosidase. Step (B) may be carried out at any timing during the production process of a beer-taste beverage, from the brewing step to the filling step, and may be carried out before, simultaneously with, or after step (A), but is preferably carried out before step (A). When step (B) is carried out, nucleosidase may be added as a raw material for the beer-taste beverage during the brewing step, or nucleosidase may be added to the wort or wort fermentation liquor at any timing during the saccharification step to the filling step. By adding nucleosidase, four types of purine nucleosides (adenosine, guanosine, inosine, and xanthosine) are hydrolyzed by the action of nucleosidase to form the purine bases (adenine, guanine, hypoxanthine, and xanthine), respectively, thereby reducing the content of the four types of nucleosides (adenosine, guanosine, inosine, and xanthosine). Meanwhile, the content of the purine bases (adenine, guanine, hypoxanthine, and xanthine) increases, but adenine, guanine, and hypoxanthine can be assimilated by common brewer's yeast, and xanthine can be decomposed in the above-mentioned step (A). Therefore, by performing step (B) before step (A), the purine content in the beer-flavored beverage can be further reduced.

[0074] In the purine reduction method, when step (B) is performed in addition to step (A), the amount of nucleosidase added is not particularly limited, but it is preferable that the amount be such that the activity of the nucleosidase is, for example, 0.01 U or more, or 0.1 U or more, or 20 U or less, or 2 U or less. In this specification, the activity of nucleosidase was defined by quantifying the amount of ribose produced by the reaction using guanosine as a substrate. Each mL of reaction solution contained 0.1 M acetate buffer (pH 4.3), 8 mM guanosine, and an appropriate amount of enzyme. The reaction was initiated by adding guanosine and allowed to proceed at 55°C for 30 minutes. The reaction was stopped by adding 1.5 mL of 0.5% dinitrosalicylic acid solution, followed by boiling for 10 minutes. The absorbance of the cooled reaction solution at 540 nm was measured, and the activity was calculated by subtracting the absorbance of the reaction solution without enzyme. The amount of enzyme required to produce 1 μmol of ribose in 30 minutes was defined as 1 U of enzyme activity.

[0075] In the purine reduction method, the above-mentioned steps (A) and (B) may be appropriately combined with other methods for reducing the purine content. Methods for reducing the purine content other than steps (A) and (B) are not particularly limited, but include, for example, methods such as adsorption and removal of purines using an adsorbent such as activated carbon, and dilution.

[0076] <<Beer-flavored beverage manufacturing method>> Next, a method for producing the beer-taste beverage of the present invention will be described. The method for producing a beer-taste beverage of the present invention (hereinafter also simply referred to as the production method of the present invention) comprises the steps of adjusting the purine content to 15 ppm or less and adjusting the content of peptides with a molecular weight of 35 to 50 kDa to 1.5 to 50 ppm. According to the production method of the present invention, it is possible to provide a beer-flavored beverage that has a reduced purine content, yet has an excellent taste and a less astringent taste.

[0077] In the production method of the present invention, the step of adjusting the purine content to 15 ppm or less is not particularly limited, but examples include a step of reducing the malt ratio, a fermentation step (purine assimilation by a microorganism such as yeast, such as xanthine assimilation by a microorganism capable of xanthine assimilation (step (A1) in the above-mentioned purine reduction method)), a xanthine decomposition step using xanthine oxidase (step (A2) in the above-mentioned purine reduction method), a hydrolysis step using nucleosidase (step (B) in the above-mentioned purine reduction method), a purine adsorption removal step using an adsorbent, and a dilution step. These steps may be performed at any timing from the brewing step to the filling step. These steps may be performed individually or in combination of two or more, as long as the purine content of the beer-taste beverage obtained by the production method of the present invention can be adjusted to 15 ppm or less. In one embodiment of the present invention, the fermentation step (assimilation of purines by microorganisms such as yeast) can be performed using, for example, a method similar to step (A1) (a step of allowing a microorganism capable of assimilating xanthine to assimilate xanthine) in the above-mentioned purine-reducing method. Furthermore, in one embodiment of the present invention, the xanthine degradation step using xanthine oxidase can be performed using, for example, a method similar to step (A2) (a step of decomposing xanthine using xanthine oxidase) in the above-mentioned purine-reducing method. Furthermore, the hydrolysis step using nucleosidase can be performed using, for example, a method similar to step (B) (a step of adding nucleosidase) in the above-mentioned purine-reducing method. Furthermore, the fermentation step (assimilation of purines by microorganisms such as yeast), the xanthine degradation step using xanthine oxidase, the hydrolysis step using nucleosidase, the step of adsorbing and removing purines using an adsorbent, the dilution step, and the like are not particularly limited, and conventionally known methods and conditions can be used. The adsorbent used for adsorbing and removing purines is not particularly limited, and examples thereof include activated carbon, zeolite, activated clay, and the like. In the production method of the present invention, the step of adjusting the purine content to 15 ppm or less is preferably step (A1) in the above-mentioned purine reduction method (the step of allowing a microorganism capable of assimilating xanthine to assimilate xanthine).

[0078] In the production method of the present invention, the step of adjusting the content of peptides with molecular weights of 35 to 50 kDa (40 kDa peptides) to 1.5 to 50 ppm includes, but is not limited to, a step of adjusting the malt ratio or the amount of soybean or yeast extract used, a hydrolysis step using peptidase, a step of adsorbing and removing the 40 kDa peptide using an adsorbent, a step of adding the 40 kDa peptide, and a dilution step. These steps may be performed at any time between the brewing step and the filling step. These steps may be performed alone or in combination of two or more, as long as the 40 kDa peptide content in the beer-taste beverage obtained by the production method of the present invention can be adjusted to 1.5 to 50 ppm. The hydrolysis step using peptidase, the adsorption and removal step of the 40 kDa peptide using an adsorbent, the step of adding the 40 kDa peptide, and the dilution step are not particularly limited, and conventionally known methods and conditions can be used. The adsorbent used for adsorbing and removing the 40 kDa peptide is not particularly limited, and examples thereof include activated carbon, zeolite, activated clay, and the like. In the production method of the present invention, the step of adjusting the content of peptides with molecular weights of 35 to 50 kDa (40 kDa peptides) to 1.5 to 50 ppm is preferably a step of adding 40 kDa peptides, from the perspective of being able to precisely adjust the content of 40 kDa peptides in the resulting beer-taste beverage, and is preferably a step of adjusting the malt ratio, from the perspective of being easy to adopt without increasing the number of steps in the production of beer-taste beverages.

[0079] In the production method of the present invention, the raw materials for the beer-taste beverage, the production process of the beer-taste beverage excluding the step of adjusting the purine content to 15 ppm or less and the step of adjusting the content of peptides with a molecular weight of 35 to 50 kDa to 1.5 to 50 ppm, the contents of each component in the beer-taste beverage obtained by the production method of the present invention, and preferred aspects thereof are the same as those for the beer-taste beverage of the present invention described above.

[0080] In this specification, a numerical range expressed by a lower limit and an upper limit, i.e., "lower limit to upper limit," includes both the lower limit and the upper limit. For example, a range expressed by "1 to 2" means 1 or more and 2 or less, including 1 and 2. In this specification, the upper and lower limits may be in any combination. [Example]

[0081] The present invention will be described in more detail below with reference to examples, but the scope of the present invention is not limited to these examples.

[0082] (Materials used) Commercially available beer-flavored beverage A (malt ratio: 50% by weight or more, sugar content: 0.3g / 100mL, alcohol content: 5.5v / v%) Activated carbon (Kawakita Chemical Co., Ltd., Kujaku Tokusen F) The commercially available beer-flavored beverage A was produced through a process of adjusting the malt ratio so that the content of peptides with a molecular weight of 35 to 50 kDa was in the range of 1.5 to 50 ppm.

[0083] <Reference example 1> A commercially available beer-taste beverage A was used as the sample for Reference Example 1.

[0084] <Reference example 2> (Activated carbon treatment) 100 mL of commercially available beer-taste beverage A was placed in an Erlenmeyer flask, 500 mg of activated carbon was added, and the mixture was stirred using a stirrer at 20°C and 500 rpm for 3 hours to adsorb the purines onto the activated carbon. The activated carbon with the adsorbed purines was then removed by filtration to obtain activated carbon-treated beer-taste beverage A. The resulting activated carbon-treated beer-taste beverage A was designated the sample of Reference Example 2.

[0085] Example 1 (Xanthine utilization reaction treatment) The yeast Torulaspora delbrueckii (LEVEL2 BIODIVA, LALLEMAND) was used as a microorganism capable of utilizing xanthine.

[0086] (preculture) The wort used for pre-culture was prepared by the following method. 50 g of ground barley malt was placed in a mash tank containing 200 mL of hot water maintained at 45°C and held there for 30 minutes. The temperature was then raised to 67°C at a rate of 1°C per minute and held there for 50 minutes. After further raising the temperature to 78°C, the mixture was filtered to remove malt grains and obtain a saccharified liquid. The resulting saccharified liquid was boiled and filtered to obtain wort. 10 mL of wort was placed in a test tube, and one loopful of Torulaspora delbrueckii was inoculated into it, followed by pre-culture under the following conditions. Culture temperature: 30℃ Culture time: 24hr Stirring: 120rpm The obtained preculture medium of Torulaspora delbrueckii was used in the following reaction.

[0087] (Xanthine utilization reaction conditions) Step (B): Nucleosidase (product name: PNF-L, manufactured by Shin-Nippon Chemical Co., Ltd., number of units: 1600 U / g) was added to a commercially available beer-flavored beverage A at 0.5 g / L and glucose at 0.5 wt % to prepare a base beer. The nucleosidase activity in the base beer was 0.8 U.

[0088] Step (A1): 250 mL of the above base beer was placed in a 1000 mL Erlenmeyer flask, and a preculture solution of Torulaspora delbrueckii was added, followed by reaction under the following conditions. Reaction temperature: 20℃ Response time: 72 hours Stirring: 120rpm After the reaction, the reaction mixture was filtered to obtain xanthine assimilation-treated beer-taste beverage A. The resulting xanthine assimilation-treated beer-taste beverage A was used as the sample of Example 1.

[0089] (Measurement of purine, 40kDa peptide, total peptide and carbohydrate content) The purine content, 40 kDa peptide content, total peptide content, and carbohydrate content were measured for beer-taste beverage A (Reference Example 1), activated carbon-treated beer-taste beverage A (Reference Example 2), and xanthine assimilation reaction-treated beer-taste beverage A (Example 1).

[0090] The purine content was measured using a liquid chromatograph-tandem mass spectrometer (LC-MS / MS) (X500R QTOF, manufactured by SCIEX) under the following conditions. Column: Discovery (registered trademark) HS F5 HPLC column (product number: 567503-U, 3 μm particle size, L × ID 15 cm × 2.1 mm, manufactured by Merck) Eluent: Gradient using 0.1% formic acid / HO (solution A) and 0.1% formic acid / acetonitrile (solution B) Gradient conditions (% is volume %): 0.00 min (Solution A:Solution B = 98%:2%), 3.00 min (Solution A:Solution B = 98%:2%), 15.00 min (Solution A:Solution B = 62%:38%), 18.00 min (Solution A:Solution B = 2%:98%), 22.00 min (Solution A:Solution B = 2%:98%), 22.01 min (Solution A:Solution B = 98%:2%), 30.00 min (Solution A:Solution B = 98%:2%) Elution rate: 0.2mL / min Column temperature: 40℃

[0091] The 40 kDa peptide content was measured by the Bradford method using a commercially available kit (Protein Assay Dye Reagent Concentrate, Bio-Rad). First, the sample was adjusted to an appropriate concentration range (0 mg / mL, 0.06125 mg / mL, 0.125 mg / mL, 0.25 mg / mL, and 0.5 mg / mL). Next, 250 μL of Protein Assay Dye Reagent Concentrate, previously diluted 5-fold with distilled water, was added to a 96-well plate. Five μL of the adjusted sample was then added to the 96-well plate, stirred, and incubated at room temperature for 30 minutes. The absorbance at 595 nm was then measured. The 40 kDa peptide content was calculated based on the absorbance and a previously prepared calibration curve. The calibration curve was prepared using bovine serum albumin (BSA).

[0092] Total peptide content was measured by the Lowry method using a commercially available kit (DC Protein Assay, Bio-Rad). First, the sample was adjusted to an appropriate concentration range (0 mg / mL, 0.06125 mg / mL, 0.125 mg / mL, 0.25 mg / mL, 0.5 mg / mL, 1 mg / mL, and 2 mg / mL). 50 μL of solution A was added to 5 μL of the adjusted sample and vortexed, followed by 400 μL of solution B. After 15 minutes of color development at room temperature, 350 μL was transferred to a 96-well plate and the absorbance at 750 nm was measured. The total peptide content was calculated based on the absorbance and a previously prepared calibration curve. The calibration curve was prepared using bovine serum albumin (BSA).

[0093] The carbohydrate content was calculated by subtracting the amounts of protein, lipid, dietary fiber, ash, and water from the weight of the beer-taste beverage. Here, the amounts of protein, lipid, dietary fiber, ash, and water were measured according to the methods set forth in the Nutrition Labeling Standards. Specifically, the amount of protein was measured using the nitrogen quantitative conversion method, the amount of lipid was measured using the ether extraction method, the amount of dietary fiber was measured using the Prosky method, the amount of ash was measured using the direct ashing method, and the amount of water was measured using the vacuum heating drying method.

[0094] (sensory evaluation) Four expert panelists evaluated the "taste" and "astringent taste" of each of the beer-taste beverage A (Reference Example 1), the activated carbon-treated beer-taste beverage A (Reference Example 2), and the xanthine assimilation-treated beer-taste beverage A (Example 1). For "taste," the sample of Reference Example 1 was assigned a score of 4.0 as the base point, and a 5% by volume aqueous ethanol solution was assigned a score of 1.0, and scores were calculated in increments of 0.1. For "astringent taste," the sample of Reference Example 1 was assigned a score of 0 as the base point, and a 0.00001% by weight aqueous quinine hydrochloride dihydrate (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) solution was assigned a score of 3, and scores were calculated in increments of 0.1. For "taste," a higher score indicates a higher rating (excellent taste), and for "astringent taste," a lower score indicates a higher rating (less astringent taste). A score of 0 for "astringency" means that no astringency was felt after drinking the beer-flavored beverage. The average scores of the four expert panelists were used as the evaluation results for "taste" and "astringency." In addition, in the "Overall Evaluation" section of Table 1, samples with a "Taste" evaluation score of 2.5 points or more and a "Astringent taste" evaluation score of less than 1.0 points were rated as "Good," and samples with a "Taste" evaluation score of less than 2.5 points and / or a "Astringent taste" evaluation score of 1.0 points or more were rated as "Poor." The results are shown in Table 1.

[0095] [Table 1]

[0096] The results of Reference Examples 1 and 2 in Table 1 confirmed that the activated carbon treatment adsorbed and removed most of the purines in commercially available beer-taste beverage A, significantly reducing the purine content. It was also confirmed that the activated carbon treatment significantly reduced the 40 kDa peptide content and total peptide content in commercially available beer-taste beverage A. As a result, the activated carbon-treated beer-taste beverage A (Reference Example 2) had unintended components, including peptides such as the 40 kDa peptide, as well as purines, adsorbed and removed by the activated carbon. As a result, the appearance and taste of the beverage resembled ethanol water, giving it a poor taste and an impression far removed from that of a beer-taste beverage.

[0097] Furthermore, the results of Reference Example 1 and Example 1 in Table 1 show that by performing the xanthine assimilation reaction treatment (step (B) of adding nucleosidase and step (A1) of allowing a microorganism capable of assimilating xanthine to assimilate xanthine), the contents of all eight purine components, including xanthine, could be reduced to below the detection limit. Furthermore, it was found that the content of components that contribute to the taste of the beer-flavored beverage (the content of the 40 kDa peptide and the total peptide content) did not decrease significantly before or after step (A1), and the taste was not significantly impaired.

[0098] <Examples 2 to 7 and Comparative Examples 1 to 4> (40kDa peptide fraction purification) (1) Fractionation by cation exchange resin 50 mL of cation exchange resin SP Sepharose was placed in an empty column and washed with 150 mL of ultrapure water, followed by 150 mL of 20 mM sodium acetate buffer (pH 4.5). 1 L of commercial beer was placed in a container, and the washed resin was added. The peptide was allowed to adsorb to the resin with occasional stirring with a spatula (1 hour). The peptide-adsorbed resin was then transferred to a column and washed with 150 mL of 20 mM sodium acetate buffer (pH 4.5). Elution was then performed with 20 mM sodium acetate (pH 4.5) + 0.5 M NaCl, and nine 50 mL fractions were collected. The resulting fractions were evaluated by SDS-PAGE, and the fraction containing the 40 kDa peptide was collected as the cation exchange resin-bound fraction. (2) Ultrafiltration (buffer exchange) 10 mL of the cation exchange resin-bound fraction obtained in (1) was added to a water-washed ultrafiltration unit (Merck Amicon Ultra-15 30 KDa), centrifuged at 3500 rpm, and ultrafiltered to obtain a concentrate. (3) Ammonium sulfate fraction A 9-fold volume of 20 mM phosphate buffer (pH 7.0) + 2 M ammonium sulfate solution was added to a beaker, and the concentrate was added dropwise and stirred for 1 hour. The suspension was then placed in a 50 mL Falcon tube and centrifuged (2330 g, 10 min). The supernatant was decanted into a separate container. The collected solution was concentrated to approximately 1 mL using an ultrafiltration unit. 10 mL of 20 mM sodium acetate (pH 4.5) was added to the concentrate, which was then centrifuged (2330 g, 10 min). This was repeated four times to obtain a purified 40 kDa peptide (Bradford assay: 18.4 mg / mL, 1.69 mL). The purity of the purified 40 kDa peptide was confirmed by SDS-PAGE.

[0099] (Preparation of a mixed solution of eight purine components in equal amounts) 6.3 mg of each of the eight components, adenosine, adenine, inosine, and hypoxanthine (all reagents manufactured by Nacalai Tesque, Inc.), and xanthine, xanthosine dihydrate, guanine, and guanosine (all reagents manufactured by Tokyo Chemical Industry Co., Ltd.), was placed in a microtube, and 500 μL of 1 mol / L NaOH aqueous solution was added and dissolved. Then, 500 μL of distilled water was added and mixed to obtain a 0.5 mol / L NaOH aqueous solution with a total content of 50,000 ppm (5 wt%) of the eight purine components. The resulting aqueous solution was used in the following examples and comparative examples as a solution containing equal amounts of the eight purine components.

[0100] (Addition of 40kDa peptide) In Examples 2 to 4 and Comparative Example 1 (Table 2), a 40 kDa peptide addition test was conducted using xanthine assimilation-treated beer-taste beverage A (Example 1) as a base. Furthermore, in Examples 5 to 7 (Table 3), a 40 kDa peptide addition test was conducted using activated carbon-treated beer-taste beverage A (Reference Example 2) as a base. Furthermore, in Comparative Examples 2 to 4 (Table 4), a 40 kDa peptide addition test was conducted using a sample (Comparative Example 2) prepared by adding an equal-volume mixed solution of eight purine components obtained by the above method to activated carbon-treated beer-taste beverage A (Reference Example 2) so that the total amount of the eight purine components added was 40 ppm. In Examples 2 to 7 and Comparative Examples 1 to 4, the purified 40 kDa peptide obtained by the above method was added to and mixed with the beer-flavored beverages used as the base for each of the above-mentioned Examples and Comparative Examples so that the 40 kDa peptide content (content in the sample) was the amount shown in Tables 2 to 4, respectively, to obtain the samples of Examples 2 to 7 and Comparative Examples 1 to 4. For the samples obtained in Examples 2 to 7 and Comparative Examples 1 to 4, the total peptide content was measured in the same manner as in Example 1.

[0101] (sensory evaluation) The samples of Examples 2 to 7 and Comparative Examples 1 to 4 were evaluated for "taste" and "astringency" using the same method and evaluation criteria as in the sensory evaluation of Example 1. In addition, an "overall evaluation" was determined based on the evaluation results of "taste" and "astringency" (average scores of four expert panelists). The results are shown in Tables 2 to 4. For comparison, Example 1 is also shown in Table 2, and Reference Example 2 is also shown in Table 3.

[0102] [Table 2]

[0103] [Table 3]

[0104] [Table 4]

[0105] <Examples 8 to 13 and Comparative Examples 5 to 7> (Peptide fraction purification) (1) Ultrafiltration (buffer exchange) 10 mL of 200 mL of commercially available beer was added to a water-washed ultrafiltration unit (Merck Amicon Ultra-15 3KDa) and centrifuged at 2330 g for 30 minutes. Centrifugation was repeated as necessary until the retentate was reduced to approximately 1 mL, and a concentrate was obtained by ultrafiltration. Next, 10 mL of 20 mM sodium acetate (pH 4.5) was added to the concentrate, which was then centrifuged (2330 g for 30 minutes) and concentrated (four times) to approximately 1 mL to obtain a peptide fraction (Lowry assay: 20.0 mg / mL, 10 mL).

[0106] (Peptide addition) In Examples 8 to 10 (Table 5), peptide addition tests were conducted using xanthine assimilation-treated beer-taste beverage A (Example 1) as a base. Furthermore, in Examples 11 to 13 and Comparative Example 5 (Table 6), peptide addition tests were conducted using activated carbon-treated beer-taste beverage A (Reference Example 2) as a base. Furthermore, in Comparative Examples 6 and 7 (Table 7), peptide addition tests were conducted using a sample (Comparative Example 2) prepared by adding an equal-volume mixed solution of eight purine components obtained by the above method to activated carbon-treated beer-taste beverage A (Reference Example 2) so that the total amount of the eight purine components added was 40 ppm. In Examples 8 to 13 and Comparative Examples 5 to 7, the peptide fractions obtained by the above-mentioned methods were added to and mixed with the beer-flavored beverages used as the base for each of the above-mentioned Examples and Comparative Examples so that the total peptide content (content in the sample) was the amount shown in Tables 5 to 7, respectively, to obtain the samples of Examples 8 to 13 and Comparative Examples 5 to 7. The content of 40 kDa peptide in the samples obtained in Examples 8 to 13 and Comparative Examples 5 to 7 was measured in the same manner as in Example 1.

[0107] (sensory evaluation) The samples of Examples 8 to 13 and Comparative Examples 5 to 7 were evaluated for "taste" and "astringency" using the same method and evaluation criteria as in the sensory evaluation of Example 1. In addition, an "overall evaluation" was determined based on the evaluation results of "taste" and "astringency" (average scores of four expert panelists). The results are shown in Tables 5 to 7. For comparison, Table 5 also shows Example 1, Table 6 also shows Reference Example 2, and Table 7 also shows Comparative Example 2.

[0108] [Table 5]

[0109] [Table 6]

[0110] [Table 7]

[0111] The results in Tables 2, 3, 5, and 6 indicate that in beer-flavored beverages with a purine content of 15 ppm or less, a 40 kDa peptide content of less than 1.5 ppm resulted in a poor taste, whereas a 40 kDa peptide content of more than 50 ppm resulted in a strong astringent taste. Furthermore, as the 40 kDa peptide content increased, the taste rating tended to improve and the astringent taste rating tended to decrease, with the astringent taste significantly worsening when the 40 kDa peptide content exceeded 50 ppm. Furthermore, comparing the results for beverages with a purine content of less than 15 ppm (Tables 3 and 6) with those with a purine content of more than 15 ppm (Tables 4 and 7), it was found that the improvement in taste due to the increased 40 kDa peptide content was more pronounced in the former case, while the degree of increase in astringent taste due to the increased 40 kDa peptide content was smaller in the former case.

Claims

1. A beer-flavored beverage having a purine content of 15 ppm or less and a peptide content of 35 to 50 kDa molecular weight of 1.5 to 50 ppm.

2. 2. The beer-taste beverage according to claim 1, wherein the content of peptides having a molecular weight of 35 to 50 kDa is 7 to 50 ppm.

3. 3. The beer-taste beverage according to claim 1 or 2, wherein the xanthine content is 5 ppm or less.

4. 3. The beer-taste beverage according to claim 1 or 2, wherein the malt content is 25% by weight or more.

5. 3. The beer-taste beverage according to claim 1 or 2, wherein the malt content is 50% by weight or more.

6. 3. The beer-taste beverage according to claim 1 or 2, wherein the carbohydrate content is less than 0.5 g / 100 mL.

7. 3. The beer-taste beverage according to claim 1 or 2, wherein the total peptide content is 60 to 600 ppm.

8. A method for producing a beer-taste beverage, comprising the steps of adjusting the purine content to 15 ppm or less and adjusting the peptide content having a molecular weight of 35 to 50 kDa to 1.5 to 50 ppm.

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