Beer-flavored beverage, method for manufacturing a beer-flavored beverage, and method for reducing purines in a beer-flavored beverage

Abstract

To provide a beer-flavored beverage with reduced high-molecular-weight purine content, a method for producing a beer-flavored beverage, and a method for reducing purines in a beer-flavored beverage. [Solution] A beer-flavored beverage having a malt ratio of 50% by weight or more, and the difference between the purine content (a) measured by method A below and the purine content (b) measured by method B below is 4.0 ppm or less. Method A: A method in which a 70% by weight perchloric acid aqueous solution is added to a beer-flavored beverage to a concentration of 18.2 v / v%, treated at 98°C for 1 hour, neutralized by adding a 4M potassium hydroxide aqueous solution to a concentration of 41 v / v%, and the purine base content in the obtained sample is measured by liquid chromatography-tandem mass spectrometry (LC-MS / MS). Method B: A method for measuring the purine base and nucleoside content in beer-flavored beverages by liquid chromatography-tandem mass spectrometry (LC-MS / MS) and calculating the sum of the purine base content and the nucleoside content in terms of purine base.

Description

Technical Field

[0001] The present invention relates to a beer-taste beverage, a method for producing a beer-taste beverage, and a method for reducing purine bodies in a beer-taste beverage.

Background Art

[0002] In recent years, due to the increasing health consciousness, the demand for purine-off or purine-free beer-taste beverages with a reduced purine body content has been increasing. Purine bodies are a general term for compounds having a purine skeleton and complexes containing compounds having a purine skeleton as components. The purine bodies contained in beer-taste beverages usually mainly include a total of eight types, namely nucleosides such as adenosine, guanosine, inosine, and xanthosine, and purine bases such as adenine, guanine, hypoxanthine, and xanthine. The content of purine bodies contained in beer-taste beverages tends to increase as the malt ratio of the beer-taste beverage increases. Therefore, in conventional purine-off or purine-free beer-taste beverages, the malt ratio is kept low, and further, if necessary, steps such as adsorption and removal of purine bodies and dilution are performed to reduce the content of purine bodies. For example, Patent Document 1 describes a method for producing a beer-taste beverage having a purine body content of 0.50 mg / 100 mL or less, in which the malt ratio is less than 5% by weight and the content of purine bodies is reduced by activated carbon treatment. Further, Patent Document 2 describes that activated clay obtained by acid-treating acid clay has excellent adsorbability to caffeine. Caffeine is a compound having a xanthine skeleton.

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Patent Document 2

Summary of the Invention

[0004] However, conventional purine-free or purine-free beer-flavored beverages have had insufficient flavor due to their low malt content (less than 50% by weight). While it is known that the flavor of beer-flavored beverages can be improved by increasing the malt content, if the malt content is increased to, for example, 50% by weight or more, the purine content inevitably increases. Thus, in order to ensure the flavor of beer-flavored beverages, the purine content in the beverage becomes high.

[0005] Furthermore, in order to ensure the taste of beer-flavored beverages, the malt ratio is set to, for example, 50% by weight or more, and in order to reduce the purine content in beer-flavored beverages, methods such as purine removal or dilution are being considered.

[0006] Here, the purines contained in beer-flavored beverages include purine bases and nucleosides that exist individually in the beer-flavored beverage (without forming complexes with other components) (hereinafter sometimes referred to as "free purines" in this specification) and complexes that include compounds having a purine skeleton as components (hereinafter sometimes referred to as "high molecular weight purines" in this specification). Therefore, in order to reduce the purine content (total content of free purines and high molecular weight purines) contained in beer-flavored beverages, reducing the content of high molecular weight purines is one useful means.

[0007] Generally, one method for reducing purine content is to remove purines contained in beer-flavored beverages by adsorption treatment with activated carbon. However, while adsorption treatment with activated carbon can reduce purine content by adsorbing and removing free purines and high molecular weight purines, it also adsorbs and removes flavor components that contribute to the taste of beer-flavored beverages, which can result in a deterioration of the taste of the beverage. Furthermore, when diluting beer-flavored beverages to reduce their purine content, there was a problem that the taste of the beer-flavored beverage became weaker and less palatable. Thus, a method for reducing the content of high-molecular-weight purines without significantly compromising the taste of beer-flavored beverages has not yet been established, and there has been a demand for beer-flavored beverages with reduced high-molecular-weight purine content. Furthermore, there have been no known cases to date where methods for specifically reducing the amount of high-molecular-weight purines contained in beer-flavored beverages have been investigated.

[0008] The present invention aims to provide a beer-flavored beverage with reduced high-molecular-weight purine content, a method for producing a beer-flavored beverage, and a method for reducing purines in a beer-flavored beverage. [Means for solving the problem]

[0009] As a result of diligent research, the inventors of the present invention discovered that by adjusting the content of high molecular weight purines in a beer-flavored beverage with a malt ratio of 50% by weight or more, a beer-flavored beverage with a reduced high molecular weight purine content can be obtained. This led to the invention of the beer-flavored beverage, a method for producing a beer-flavored beverage, and a method for reducing purines in a beer-flavored beverage.

[0010] In other words, the present invention relates to the following beer-flavored beverages, etc., although it is not limited to the following. [1] A beer-flavored beverage having a malt ratio of 50% by weight or more, and the difference between the purine content measured by method A below (a) and the purine content measured by method B below (b) is 4.0 ppm or less. Method A: A method in which a 70% by weight perchloric acid aqueous solution is added to a beer-flavored beverage to a concentration of 18.2 v / v%, treated at 98°C for 1 hour, neutralized by adding a 4M potassium hydroxide aqueous solution to a concentration of 41 v / v%, and the purine base content in the obtained sample is measured by liquid chromatography-tandem mass spectrometry (LC-MS / MS). Method B: A method for measuring the purine base and nucleoside content in beer-flavored beverages by liquid chromatography-tandem mass spectrometry (LC-MS / MS) and calculating the sum of the purine base content and the nucleoside content in terms of purine base. [2] The beer-flavored beverage described in [1] above, wherein the purine content (a) is less than 5.0 ppm. [3] A beer-flavored beverage as described in [1] or [2] above, wherein the real extract concentration is 0.5% by weight or more. [4] A beer-flavored beverage according to any of [1] to [3] above, having a total peptide content of 60 ppm or more. [5] A beer-flavored beverage as described in any of [1] to [4] above, having a total polyphenol content of 50 ppm or more. [6] A beer-flavored beverage as described in any of [1] to [5] above, having a carbohydrate content of less than 0.5g / 100mL. [7] A method for producing a beer-flavored beverage having a malt ratio of 50% by weight or more, comprising a purine content adjustment step of adjusting the difference between the purine content (a) measured by the method A below and the purine content (b) measured by the method B below to 4.0 ppm or less. Method A: A method in which a 70% by weight perchloric acid aqueous solution is added to a beer-flavored beverage to a concentration of 18.2 v / v%, treated at 98°C for 1 hour, neutralized by adding a 4M potassium hydroxide aqueous solution to a concentration of 41 v / v%, and the purine base content in the obtained sample is measured by liquid chromatography-tandem mass spectrometry (LC-MS / MS). Method B: A method for measuring the purine base and nucleoside content in beer-flavored beverages by liquid chromatography-tandem mass spectrometry (LC-MS / MS) and calculating the sum of the purine base content and the nucleoside content in terms of purine base. [8] The method for producing a beer-flavored beverage as described in [7] above, wherein the purine content adjustment step includes an adsorption step of adsorbing purines contained in the wort or wort fermentation liquid onto a cation exchange resin. [9] The method for producing a beer-flavored beverage as described in [8] above, wherein the amount of cation exchange resin used relative to the wort or wort ferment liquor in the adsorption step is 500 to 4000 ppm.

[10] A method for producing a beer-flavored beverage according to [8] or [9] above, wherein the purine content adjustment step further includes a xanthine utilization step, prior to the adsorption step, in which xanthine contained in the wort or wort ferment liquor is utilized by yeast having xanthine utilization ability.

[11] The method for producing a beer-flavored beverage according to

[10] above, wherein the purine content adjustment step further comprises a nucleosidase treatment step of treating the wort or wort ferment with nucleosidase before the xanthine assimilation step.

[12] A method for producing a beer-flavored beverage according to any one of [8] to

[11] above, wherein the wort fermentation liquid has a ratio of xanthine content to the total content of adenine and guanine (xanthine content / total content of adenine and guanine) of less than 0.5.

[13] A method for producing a beer-flavored beverage according to any one of [7] to

[12] above, wherein in the purine content adjustment step, the purine content (a) is adjusted to 5.0 ppm or less.

[14] A method for reducing the purine content of a beer-flavored beverage having a malt ratio of 50% by weight or more, comprising a purine content adjustment step of adjusting the difference between the purine content (a) measured by the method A below and the purine content (b) measured by the method B below to 4.0 ppm or less. Method A: A method in which a 70% by weight perchloric acid aqueous solution is added to a beer-flavored beverage to a concentration of 18.2 v / v%, treated at 98°C for 1 hour, neutralized by adding a 4M potassium hydroxide aqueous solution to a concentration of 41 v / v%, and the purine base content in the obtained sample is measured by liquid chromatography-tandem mass spectrometry (LC-MS / MS). Method B: A method for measuring the purine base and nucleoside content in beer-flavored beverages by liquid chromatography-tandem mass spectrometry (LC-MS / MS) and calculating the sum of the purine base content and the nucleoside content in terms of purine base.

[15] The method for reducing the purine content of a beer-flavored beverage as described in

[14] above, wherein the purine content adjustment step includes an adsorption step of adsorbing purines contained in the wort or wort fermentation liquid onto a cation exchange resin.

[16] The method for reducing purines in a beer-flavored beverage according to

[15] above, wherein the amount of cation exchange resin used relative to the wort or wort ferment liquor in the adsorption step is 500 to 4000 ppm.

[17] The method for reducing purines in a beer-flavored beverage according to

[15] or

[16] above, wherein the purine content adjustment step further includes a xanthine utilization step, prior to the adsorption step, in which xanthine contained in the wort or wort ferment liquor is utilized by yeast having xanthine utilization ability.

[18] The method for reducing purines in a beer-flavored beverage according to

[17] above, wherein the purine content adjustment step further includes a nucleosidase treatment step of treating the wort or wort ferment with nucleosidase before the xanthine assimilation step.

[19] A method for reducing purines in a beer-flavored beverage according to any of

[15] to

[18] above, wherein the ratio of the xanthine content to the total adenine and guanine content (xanthine content / total adenine and guanine content) in the wort ferment is less than 0.5.

[20] A method for reducing the purine content of a beer-flavored beverage according to any one of

[14] to

[19] above, wherein in the purine content adjustment step, the purine content (a) is adjusted to 5.0 ppm or less. [Effects of the Invention]

[0011] The beer - flavored beverage of the present invention has a malt ratio of 50% by weight or more and a reduced content of high - molecular - weight purine bodies, so it is useful from the perspective of consumers' health. Also, according to the method for producing the beer - flavored beverage of the present invention, a beer - flavored beverage with a reduced content of high - molecular - weight purine bodies can be provided. Furthermore, according to the method for reducing purine bodies in the beer - flavored beverage of the present invention, it is possible to reduce the content of high - molecular - weight purine bodies in the beer - flavored beverage.

Embodiments for Carrying Out the Invention

[0012] <<Beer - flavored Beverage>> First, the beer - flavored beverage of the present invention will be described. The beer - flavored beverage of the present invention has a malt ratio of 50% by weight or more, and the difference between the content (a) of purine bodies measured by the following method A and the content (b) of purine bodies measured by the following method B is 4.0 ppm or less, which is a beer - flavored beverage. Method A: Add 70% by weight perchloric acid aqueous solution to the beer - flavored beverage so that it becomes 18.2 v / v%, treat it at 98°C for 1 hour, then add 4M potassium hydroxide aqueous solution so that it becomes 41 v / v% for neutralization, and measure the content of purine bases in the sample obtained by recovering the supernatant by liquid chromatography tandem mass spectrometry (LC - MS / MS). Method B: Measure the content of purine bases and nucleosides in the beer - flavored beverage by liquid chromatography tandem mass spectrometry (LC - MS / MS), and calculate the sum of the content of purine bases and the content of nucleosides converted to purine bases. The beer - flavored beverage of the present invention has a malt ratio of 50% by weight or more and a reduced content of high - molecular - weight purine bodies, so it is useful from the perspective of consumers' health. In the present invention, as an index for the content of high - molecular - weight purine bodies, the "content of high - molecular - weight purine bodies converted to purine bases" described later is used.

[0013] In this specification, "beer-flavored beverage" means an alcoholic or non-alcoholic carbonated beverage that has a beer-like flavor. In other words, unless otherwise specified, "beer-flavored beverage" in this specification includes any carbonated beverage that has a beer flavor. Therefore, "beer-flavored beverages" include not only beer, which is a malt-fermented beverage obtained by fermenting malt, hops, and water using yeast, and fermented beer-flavored beverages, but also carbonated beverages to which beer flavorings containing esters, higher alcohols, lactones, etc., have been added. One embodiment of the present invention is a beer-flavored beverage, which is beer. Furthermore, "beer-flavored beverage" may be a fermented beer-flavored beverage that has undergone a fermentation process using yeast, or it may be a non-fermented beer-flavored beverage that has not undergone a fermentation process. Furthermore, the "beer-flavored beverage" may also be a beer-flavored beverage containing distilled spirits such as spirits, whiskey, or shochu. Among these, a beer-flavored beverage containing spirits is preferred.

[0014] In this specification, "flavor" refers to the depth and complexity of flavor characteristic of beer-flavored beverages. Furthermore, in this specification, the presence and degree of "flavor" can be evaluated by sensory evaluation by a panel of experts.

[0015] The beer-flavored beverage of the present invention has a malt ratio of 50% by weight or more. The malt ratio of the beer-flavored beverage of the present invention is not particularly limited as long as it is 50% by weight or more, and may be 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, and may also 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 the value calculated in accordance with the Liquor Tax Act and the Interpretation Circular on Laws and Regulations Related to the Administration of Liquor, effective April 1, 2018.

[0016] In this specification, "malt" refers to germinated and dried seeds of grains such as barley, wheat, rye, oats, oats, pearl oats, and oats, with the roots removed, and the origin and variety may be any of them. Barley malt is one of the most commonly used malts as an ingredient in Japanese beer-flavored beverages. There are various types of barley, such as two-row barley and six-row barley, and any type can be used. In addition to regular malt, colored malts can also be used. When using colored malts, different types of colored malts may be used in appropriate combinations, or a single type of colored malt may be used. In the beer-flavored beverage of the present invention, the malt is preferably barley malt.

[0017] The beer-flavored beverage of the present invention may use ingredients other than malt, such as grains other than malt, protein, or sugar solution, as long as the malt ratio is 50% by weight or more. Other grains besides malt include, for example, grains that are not malted (barley, wheat, rye, oats, oats, pearl oats, etc.), rice (white rice, brown rice, etc.), corn (corn grits, etc.), sorghum, potatoes, beans (soybeans, peas, etc.), buckwheat, sorghum, millet, barnyard millet, and starches obtained from them, as well as extracts thereof. Among these, those using corn (corn grits, etc.) are preferred. As for proteins, examples include soy protein, pea protein, and their hydrolysates.

[0018] The beer-flavored beverage of the present invention has a difference of 4.0 ppm or less between the purine content (a) measured by the following method A and the purine content (b) measured by the following method B. Method A: A method in which a 70% by weight perchloric acid aqueous solution is added to a beer-flavored beverage to a concentration of 18.2 v / v%, treated at 98°C for 1 hour, neutralized by adding a 4M potassium hydroxide aqueous solution to a concentration of 41 v / v%, and the purine base content in the obtained sample is measured by liquid chromatography-tandem mass spectrometry (LC-MS / MS). Method B: A method for measuring the purine base and nucleoside content in beer-flavored beverages by liquid chromatography-tandem mass spectrometry (LC-MS / MS) and calculating the sum of the purine base content and the nucleoside content in terms of purine base.

[0019] In this specification, "purine" refers to a general term for compounds having a purine skeleton ("free purines," described later) and complexes containing compounds having a purine skeleton as components ("high molecular weight purines," described later). Examples of compounds having a purine skeleton include purine bases, nucleosides, nucleotides, nucleic acids, etc. Examples of purine bases include adenine, guanine, hypoxanthine, and xanthine. Examples of nucleosides include adenosine, guanosine, inosine, and xanthosine, which have a structure in which ribose is bonded to one of the above purine bases (adenine, guanine, hypoxanthine, and xanthine, respectively). Examples of nucleotides include compounds that have a structure in which a phosphate group is further bonded to the above nucleosides, such as ATP, ADP, AMP, GTP, GDP, and GMP. Examples of nucleic acids include RNA and DNA.

[0020] In this specification, purine bases and nucleosides present in beer-flavored beverages individually (without forming complexes with other components) are referred to as "free purines." In this specification, "free purine content in terms of purine bases" is the sum of the content of purine bases present individually in the beer-flavored beverage and the content of nucleosides present individually in the beer-flavored beverage in terms of purine bases. The "free purine content in terms of purine bases" does not include the weight of ribose contained in the nucleosides. In this specification, the purine content of nucleosides means the value obtained by multiplying the weight of the nucleoside by the molecular weight ratio of the corresponding purine base to the nucleoside (molecular weight of purine base / molecular weight of nucleoside). The purine bases corresponding to adenosine, guanosine, inosine, and xanthosine are adenine, guanine, hypoxanthine, and xanthine, respectively. Since nucleotides and nucleic acids typically exist in beer-flavored beverages in extremely small amounts, this specification does not consider them and defines only purine bases and nucleosides present in beer-flavored beverages as "free purines," considering the "content of free purines in terms of purine bases."

[0021] In this specification, "high molecular weight purine" refers to a complex containing a compound having a purine skeleton as a component. In this specification, "purine base equivalent content of high molecular weight purines" can also be expressed as "purine base equivalent content of compounds having a purine skeleton that constitute high molecular weight purines," and refers to the purine base equivalent content of nucleosides and purine bases that exist in the beer-flavored beverage in the form of complexes with other components. In other words, "purine base equivalent content of high molecular weight purines" is the sum of the purine base equivalent content of purine bases that exist in the beer-flavored beverage in the form of complexes with other components and the purine base equivalent content of nucleosides that exist in the beer-flavored beverage in the form of complexes with other components. "Purine base equivalent content of high molecular weight purines" does not include the weight of other components that make up the complex or the weight of ribose contained in the nucleosides.

[0022] Method A described above is a method in which high molecular weight purines and nucleosides contained in a beer-flavored beverage are decomposed by perchloric acid treatment to produce purine bases, and the purine content (a), which is the sum of the amount of free purines contained in the beer-flavored beverage before perchloric acid treatment in terms of purine bases (amount of purine bases produced by the decomposition of nucleosides by perchloric acid treatment) and the amount of purine bases produced by the decomposition of high molecular weight purines by perchloric acid treatment, is measured by liquid chromatography-tandem mass spectrometry (LC-MS / MS). Notwithstanding the present invention, when "purine content" is used in the field of beer-flavored beverages in general, it usually refers to the purine content (a) measured by the above method A.

[0023] Method B described above is a method for measuring and calculating the amount of free purines in beer-flavored beverages in terms of purine bases (purine content (b)) by liquid chromatography-tandem mass spectrometry (LC-MS / MS). The measurement conditions for LC-MS / MS can be those described in the examples. By subtracting the purine content (a) obtained by method (A) from the purine content (b) obtained by method (A) above, the "purine content (b)" of free purines measured by method B above can be used to calculate the "purine content (b)" of high molecular weight purines contained in beer-flavored beverages. In other words, the "purine content (b)" of high molecular weight purines can be calculated using the following formula. (Purine base content of high molecular weight purines) = (Purine content (a)) - (Purine content (b))

[0024] In this specification, the measurement of free purines using a liquid chromatograph-tandem mass spectrometer (LC-MS / MS) can be performed in accordance with the method described in "Guidance on Trace Analysis of Purines in Alcoholic Beverages" (Japan Food Research Laboratories, Internet (https: / / www.jfrl.or.jp / storage / file / news_vol4_no23.pdf), accessed March 2024).

[0025] In method A described above, the method and conditions for collecting the supernatant are not particularly limited, and conventionally known methods and conditions can be used. For example, the supernatant can be collected by centrifuging using a centrifuge at 100 to 3000 G for 1 to 120 minutes. Alternatively, the supernatant can be collected by visually confirming the sedimentation of precipitated salts by allowing the neutralized solution to stand.

[0026] In the beer-flavored beverage of the present invention, the content of high molecular weight purines in terms of purine bases is not particularly limited as long as it is 4.0 ppm or less, but from a health-conscious viewpoint, it is preferable that it be 3.9 ppm or less, 3.8 ppm or less, 3.7 ppm or less, 3.6 ppm or less, 3.5 ppm or less, 3.4 ppm or less, 3.3 ppm or less, or 3.2 ppm or less. The lower limit of the content of high molecular weight purines in terms of purine bases is not particularly limited, but for example, it can be 0 ppm, 0.01 ppm, or 0.1 ppm.

[0027] In the beer-flavored beverage of the present invention, the purine content (a) measured by the above method A is not particularly limited, but from a health-conscious viewpoint, it is preferable to be 14.0 ppm or less, 13.0 ppm or less, 12.0 ppm or less, 11.0 ppm or less, 10.0 ppm or less, 9.0 ppm or less, 8.0 ppm or less, 7.0 ppm or less, 6.0 ppm or less, 5.0 ppm or less, or less than 5.0 ppm. The lower limit of the purine content (a) is not particularly limited, but for example, it can be 0 ppm, 0.01 ppm, 0.02 ppm, 0.03 ppm, 0.04 ppm, 0.05 ppm, 0.06 ppm, 0.07 ppm, 0.08 ppm, 0.09 ppm, or 0.1 ppm. The beer-flavored beverage of the present invention may be a low-purine beer-flavored beverage, a purine-free beer-flavored beverage, or a purine-free beer-flavored beverage. From a health-conscious standpoint, it is preferable that the purine content (a) of the beer-flavored beverage of the present invention is less than 5.0 ppm.

[0028] In this specification, "ppm" means "ppm by weight," and 10 -4 This expresses a percentage by weight. Also, unless otherwise specified, "mg / 100mL" means 10 -3 This is expressed as a percentage by weight. That is, 1 ppm = 1 mg / L = 0.1 mg / 100 mL = 0.0001 wt% = 1 μg / mL.

[0029] In the beer-flavored beverage of the present invention, the real extract concentration is not particularly limited, but is preferably 0.1% by weight or more, 0.2% by weight or more, 0.3% by weight or more, 0.4% by weight or more, 0.5% by weight or more, 0.6% by weight or more, 0.7% by weight or more, 0.8% by weight or more, or 0.9% by weight or more. The upper limit of the real extract concentration is not particularly limited, but can be, for example, 4.0% by weight or less, 3.5% by weight or less, 3.0% by weight or less, 2.5% by weight or less, 2.0% by weight or less, 1.8% by weight or less, 1.6% by weight or less, 1.5% by weight or less, 1.4% by weight or less, 1.3% by weight or less, 1.2% by weight or less, 1.1% by weight or less, or 1.0% by weight or less.

[0030] In this specification, "real extract" is synonymous with "genuine extract." The real extract concentration can be measured according to section 8.4.3, "Alcoholizer Method," of the literature "Revised BCOJ Beer Analysis Methods 2013 Supplementary and Revised Edition (Edited by: International Technical Committee (Analysis Committee) of the Beer Brewers Association, Published by: Japan Brewing Association)." The adjustment of the real extract concentration depends on the addition of dilution water or carbonated water, the type of raw materials (malt, corn grits, sugar solution, etc.), the particle size of the malt, the method of malt grinding (wet grinding, dry grinding, etc.), the humidity during malt grinding (degree of humidity control), the temperature during malt grinding, the type of mill used for malt grinding, the amount of raw materials, the type of enzyme, the amount of enzyme added, the timing of enzyme addition, the time of enzymatic decomposition, the saccharification time in the mashing tank, the protein decomposition time in the mashing tank, the pH in the mashing tank, the pH during the mashing process (the wort production process from malt addition to before yeast addition), the amount of acid added for pH adjustment, and the timing of pH adjustment (at mashing). The process can be carried out by appropriately setting the temperature and holding time for each temperature range when preparing the wort (including during saccharification), the wort filtration time, the temperature during wort filtration, the pH during wort filtration, the amount of wort recovered during wort filtration, the amount of sparging water during wort filtration, the pH of the sparging water during wort filtration, the temperature of the sparging water during wort filtration, the fermentation conditions (oxygen concentration, aeration conditions, yeast variety, amount of yeast added, yeast growth rate, yeast removal timing, fermentation temperature, fermentation time, pressure setting, carbon dioxide concentration, etc.), and the addition of spirits or brewing alcohol, etc.

[0031] The beer-flavored beverage of the present invention may be an alcohol-containing beer-flavored beverage with an alcohol content of 1(v / v)% or more, or it may be a non-alcoholic beer-flavored beverage with an alcohol content of less than 1(v / v). The beer-flavored beverage of the present invention is preferably an alcohol-containing beer-flavored beverage.

[0032] In this specification, "alcohol-containing beer-flavored beverage" refers to a beer-flavored beverage with an alcohol content of 1(v / v)% or higher. The origin of the alcohol contained in an alcohol-containing beer-flavored beverage is not limited to fermentation or non-fermentation.

[0033] In this specification, "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 a pre-fermentation liquid containing wort or components necessary for fermentation, but also include fermented and non-fermented carbonated beverages to which beer flavorings (flavorings that evoke a beer-like aroma) are added, such as esters, higher alcohols, and lactones, for example, 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, myrcene, etc. A "non-alcoholic beer-flavored beverage" may be a fermented beverage in which, after undergoing a fermentation process using yeast (top-fermenting yeast and / or bottom-fermenting yeast) in the manufacturing process, the alcohol produced in the fermentation process is removed, and the resulting alcohol content is less than 1 (v / v%). It may also be a fermented beverage obtained by stopping fermentation so that the alcohol content is less than 1 (v / v%). Alternatively, it may be a fermented beverage diluted with water or other liquids so that the alcohol content is less than 1 (v / v%). When stopping fermentation, it is preferable to stop fermentation so that off-flavors such as hydrogen sulfide, diacetyl, 2,3-pentanedione, and acetaldehyde are below a threshold, but it is not necessarily required to keep them below the threshold. The concentration of off-flavors is not limited as long as the off-flavors such as hydrogen sulfide, diacetyl, 2,3-pentanedione, and acetaldehyde blend with the flavor of the beer-flavored beverage to create a good flavor. A "non-alcoholic beer-flavored beverage" may also be a non-fermented beverage prepared without undergoing a fermentation process.

[0034] In this specification, the alcohol content of non-alcoholic beer-flavored beverages 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. Examples of non-alcoholic beer-flavored beverages include non-alcoholic beer-flavored beverages and beer-flavored soft drinks. In this specification, "alcohol content" refers to the ethanol content ((v / v)%) and does not include the content of aliphatic alcohols other than ethanol. In this specification, "volume %" and "(v / v)%" are synonymous.

[0035] For the production of alcohol-containing beer-flavored beverages and non-alcoholic beer-flavored beverages, yeast may be used to produce fermented beer-flavored beverages. Fermented beer-flavored beverages may be top-fermented beer-flavored beverages (ale-flavored beverages) brewed through a fermentation process using top-fermenting yeast (such as Saccharomyces), or bottom-fermented beer-flavored beverages (lager-flavored beverages, pilsner-flavored beverages) brewed through a fermentation process using bottom-fermenting yeast (such as Saccharomyces), and these may be blended. For fermentation, yeast that produces alcohol (Saccharomyces) or wild yeast (such as Brettanomyces) may be used, or yeast that does not produce alcohol (such as Saccharomyces), wild yeast (such as Brettanomyces), or bacteria that perform lactic acid fermentation or gluconic acid fermentation may be used.

[0036] Alcohol content shall be expressed as a percentage of volume / volume ((v / v)%). The alcohol content of a beverage may be measured by any known method, but for example, it may be measured according to "8.3 Alcohol" in the "Revised BCOJ Beer Analysis Method 2013 Supplementary Revision (edited by the International Technical Committee (Analysis Committee) of the Beer Brewers Association, published by the Japan Brewing Association)."

[0037] Alcohol content adjustment is controlled by the addition of dilution water or carbonated water, the type and amount of raw materials (malt, corn grits, sugar solution, etc.), the type of enzyme, the amount of enzyme added, the timing of enzyme addition, the saccharification time in the mashing tank, the protein decomposition time in the mashing tank, the pH in the mashing tank, the pH during the mashing process (from malt addition to wort production before yeast addition), the amount of acid added for pH adjustment, the timing of pH adjustment (during mashing, fermentation, completion of fermentation, before beer filtration, after beer filtration, etc.), and the temperature during wort preparation (including saccharification). The process can be carried out by appropriately setting the temperature and holding time of the region, the original extract concentration of the liquid before fermentation, the original extract concentration during the fermentation process, fermentation conditions (oxygen concentration, aeration conditions, yeast variety, amount of yeast added, number of yeast cells, timing of yeast removal, fermentation temperature, fermentation time, pressure setting, carbon dioxide concentration, etc.), ethanol addition, and ethanol composition (raw material alcohol, shochu, awamori, whiskey, brandy, vodka, rum, tequila, gin and other spirits (distilled liquor), brewing alcohol, etc.).

[0038] In the case where the beer-flavored beverage of the present invention is an alcohol-containing beer-flavored beverage, the alcohol content is not particularly limited as long as it is 1% by volume or more, and is not limited to 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. Preferably, the amount is 4.9 volume% or more, 5.0 volume% or more, 5.1 volume% or more, 5.2 volume% or more, 5.3 volume% or more, 5.4 volume% or more, 5.5 volume% or more, 5.6 volume% or more, 5.7 volume% or more, 5.8 volume% or more, 5.9 volume% or more, 6.0 volume% or more, 6.1 volume% or more, 6.2 volume% or more, 6.3 volume% or more, 6.4 volume% or more, 6.5 volume% or more, 7.0 volume% or more, 7.5 volume% or more, 8.0 volume% or more, 8.5 volume% or more, 9.0 volume% or more, 9.5 volume% or more, or 10.0 volume% or more. Furthermore, it is preferable that the amount is 20.0% by volume or less, 19.5% by volume or less, 19.0% by volume or less, 18.5% by volume or less, 18.0% by volume or less, 17.5% by volume or less, 17.0% by volume or less, 16.5% by volume or less, 16.0% by volume or less, 15.5% by volume or less, 15.0% by volume or less, 14.5% by volume or less, 14.0% by volume or less, 13.5% by volume or less, 13.0% by volume or less, 12.5% ​​by volume or less, 12.0% by volume or less, 11.5% by volume or less, 11.0% by volume or less, 10.5% by volume or less, or 10% by volume or less.

[0039] The following shows the manufacturing process for a typical beer-flavored beverage. First, a mixture containing malt and other grains, along with other grains as needed, starch, sugars, bittering agents, or coloring agents, and water, is mixed with enzymes such as amylase as needed to perform gelatinization and saccharification, then filtered to obtain a saccharified liquid. Hops and bittering agents are added to the saccharified liquid as needed and boiled, and solids such as coagulated proteins are removed in a clarification tank. As an alternative to this saccharified liquid, malt extract may be mixed with warm water and hops added and boiled. Hops may be added at any stage from the start of boiling to before the end of boiling. Known conditions may be used for the saccharification process, boiling process, solids removal process, fermentation and storage process, etc. The obtained fermented liquid is filtered, and carbon dioxide is added to the resulting filtrate. Then, it is filled into containers and subjected to a sterilization process to obtain the desired beer-flavored beverage.

[0040] The beer-flavored beverage of the present invention may or may not use hops as an ingredient. Examples of hop forms include pelletized hops, powdered hops, and hop extract. Alternatively, processed hop products such as isopropyl hops and reduced hops may be used as hops. Furthermore, when hops are used as an ingredient, the amount of hops used is not particularly limited, but typically it is about 0.0001 to 1% by weight of the total amount of beer-flavored beverage.

[0041] The beer-flavored beverage of the present invention may contain various additives as needed, as long as they do not interfere with the effects of the present invention. Examples of such additives include sweeteners (including high-intensity sweeteners), bittering agents or bittering agents, flavorings, colorings, foaming agents, fermentation accelerators, protein-based substances such as peptide-containing substances, dietary fiber, seasonings such as amino acids, antioxidants, and other additives. These may be used individually or in combination of two or more.

[0042] Examples of sweeteners include commercially available saccharified saccharified syrups obtained by decomposing grain-derived starch with acid or enzymes, sucrose, commercially available corn syrup and other sugars, trisaccharides or higher sugars, sugar alcohols, isomerized sugars, natural sweeteners such as stevia, and artificial sweeteners. These sugars may be in liquid form such as solutions, or in solid form such as powders. There are no particular restrictions on the type of grain used as the raw material for starch, the method of refining the starch, or the processing conditions such as hydrolysis with enzymes or acids. For example, sugars with a higher proportion of maltose may be used by appropriately setting the conditions for hydrolysis with enzymes or acids. Other examples of sweeteners that can be used include sucrose, fructose, glucose, maltose, trehalose, maltotriose, maltotetraose, isomaltose, isomalttriose, isomalttetraose, and solutions (sugar solutions) of these. Examples of artificial sweeteners include aspartame, acesulfame potassium (acesulfame K), sucralose, and neotame. These sweeteners may be used individually or in combination of two or more.

[0043] The bittering agents or bittering fertilizers are not particularly limited and include, in addition to hops, those commonly used as bittering fertilizers in beer and sparkling wine, such as rosemary, lychee, fennel, juniper berries, sage, maize, Ganoderma lucidum, bay laurel, kwashin, citrus extracts, turban extracts, coffee extracts, tea extracts, bitter melon extracts, lotus germ extracts, aloe vera extracts, rosemary extracts, lychee extracts, laurel extracts, sage extracts, caraway extracts, naringin, wormwood and wormwood extracts, absinthin, alginic acid, etc. These bittering agents or bittering fertilizers may be used individually or in combination of two or more.

[0044] The flavorings used are not particularly limited, and general beer flavorings can be used. Beer flavorings are used to give a beer-like flavor and include brewing components that are produced during 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, ethylmaltol, phenylacetic acid, furaneol, furfural, methional, 3-methyl-2-buten-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-butyrol Lactone, 2-aminoacetophenone, ethyl 3-phenylpropionate, 2-ethyl-4-hydroxy-5-methyl-3(2H)-furanone, dimethyl sulfone, 3-methylcyclopentan-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-U 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, p-methyl tolulate, 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, Ethylheptanoate, Isobutyl acetate, Activated amyl acetate, Propion Examples include ethyl acid, 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, linalool oxide, etc. These fragrances may be used individually or in combination of two or more.

[0045] Colorants used to give beverages a beer-like color include, for example, caramel color, lycopene color, elderberry color, cocoa color, and safflower color. These colorants may be used individually or in combination of two or more.

[0046] Foam-forming agents are used to create beer-like foam in beverages or to retain foam in beverages. Examples include plant-derived saponin substances such as soy saponin and quillaja saponin, plant proteins such as corn and soy, peptide-containing substances such as collagen peptides, and raw materials derived from milk. These foam-forming agents may be used individually or in combination of two or more.

[0047] Fermentation accelerators are used to promote fermentation by yeast, and examples include rice bran components, vitamins, and minerals. These fermentation accelerators may be used individually or in combination of two or more.

[0048] Examples of dietary fiber include water-soluble dietary fiber. Examples of water-soluble dietary fiber 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 are preferred. These dietary fibers may be used individually or in combination of two or more.

[0049] The antioxidants used are not particularly limited and include those commonly used as antioxidants in regular beer and sparkling wine, such as ascorbic acid, erythorbic acid, and catechin. These antioxidants may be used individually or in combination of two or more.

[0050] 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, sansho pepper, and other spices or their raw materials; chamomile, sage, basil, lemongrass, and other herbs; sweet potatoes, pumpkins, and other vegetables (including dried or boiled vegetables); buckwheat or sesame; honey or other sugary substances, salt or miso; flowers or tea, coffee, cocoa, or preparations thereof; oysters, kelp, wakame seaweed, or bonito flakes. These other additives may be used individually or in combination of two or more. If the beer-flavored beverage of the present invention contains other additives, the amount is not particularly limited, but is usually 5% by weight or less based on the weight of malt used as a raw material for the beer-flavored beverage.

[0051] In the production of beer-flavored beverages, by keeping the malt ratio low, and by performing processes such as adsorption and removal of high molecular weight purines using adsorbents such as cation exchange resins and activated carbon (adsorption process described later), fermentation (assimilation of free purines such as xanthine assimilation by yeast with xanthine assimilation ability (xanthine assimilation process described later)), decomposition of xanthine using xanthine oxidase (xanthine oxidase treatment process described later), hydrolysis using nucleosidase (nucleosidase treatment process described later), adsorption and removal of free purines using adsorbents such as activated clay and zeolite, and dilution at any time between the mashing and filling processes, a beer-flavored beverage with reduced purine content (a) can be obtained. Furthermore, by performing adsorption and removal of high molecular weight purines using adsorbents such as cation exchange resins and activated carbon (adsorption process described later) at any time between the mashing and filling processes, the content of high molecular weight purines can be reduced, resulting in a beer-flavored beverage with reduced purine content (a). These can be performed individually or in combination of two or more types. In one embodiment of the present invention, as the adsorption and removal of high molecular weight purines using adsorbents such as cation exchange resins and activated carbon, for example, a method similar to the adsorption step described later can be employed. Also in one embodiment of the present invention, as the fermentation (purine assimilation by yeast) step, for example, a method similar to the xanthine assimilation step (a step in which yeast having xanthine assimilation ability assimilates xanthine) described later can be employed. Also in one embodiment of the present invention, as the xanthine decomposition step using xanthine oxidase, for example, a method similar to the xanthine oxidase treatment step described later can be employed. Also, as the hydrolysis step using nucleosidase, for example, a method similar to the nucleosidase treatment step described later can be employed. Furthermore, the steps such as fermentation (assimilation of free purines by yeast), xanthine decomposition using xanthine oxidase, hydrolysis using nucleosidase, adsorption and removal of free purines using adsorbents such as activated clay and zeolite are not particularly limited, and conventionally known methods and conditions can be employed. The adsorbent used for the adsorption and removal of free purines is not particularly limited and includes activated carbon, zeolite, activated clay, and the like.

[0052] The beer-flavored beverage of the present invention is preferably manufactured by the method for manufacturing a beer-flavored beverage described later. Furthermore, the beer-flavored beverage of the present invention is preferably manufactured after undergoing the purine reduction method described later.

[0053] When the beer-flavored beverage of the present invention is an alcohol-containing beer-flavored beverage, the alcohol content of the final product can be adjusted by adding raw material alcohol or the like. The addition of raw material alcohol may be carried out at any stage from the saccharification process to the filling process.

[0054] From the viewpoint of imparting an alcoholic taste to the beer-flavored beverage of the present invention, an aliphatic alcohol may be added. The aliphatic alcohol is not particularly limited as long as it is known, but aliphatic alcohols having 4 to 5 carbon atoms are 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 individually or in combination of two or more.

[0055] In the beer-flavored beverage of the present invention, the content of aliphatic alcohols 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 aliphatic alcohols having 4 to 5 carbon atoms can be measured using headspace gas chromatography.

[0056] The beer-flavored beverage of the present invention may be a low-sugar beverage or a sugar-free beverage, in line with the recent trend towards low-sugar products. In the beer-flavored beverage of the present invention, the carbohydrate content can be set according to the characteristics to be imparted to the beverage, for example, 2.0g / 100mL or less, 1.9g / 100mL or less, 1.8g / 100mL or less, 1.7g / 100mL or less, 1.6g / 100mL or less, 1.5g / 100mL or less, less than 1.5g / 100mL, 1.4g / 100mL or less, 1.3g / 100mL 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 also be 100mL or less, 0.5g / 100mL or less, or less than 0.5g / 100mL, or 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, greater than 0.5g / 100mL, 0.55g / 100mL or more, 0.6g / 100mL or more, 0.65g / 100mL or more, 0. It may also 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-flavored beverage of the present invention preferably has a carbohydrate content of less than 0.5g / 100mL.

[0057] In this specification, "carbohydrates" refers to carbohydrates as defined in the Food Nutrition Labeling Standards (Ministry of Health, Labour and Welfare Notification No. 176 of 2003, partially amended by Consumer Affairs Agency Notification No. 8 of September 27, 2013), and specifically means what remains after removing protein, lipids, dietary fiber, ash, alcohol, and water from the food in question. The carbohydrate content in food can be calculated by subtracting the amounts of protein, fat, dietary fiber, ash, and water from the weight of the food. Here, the amounts of protein, lipids, 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 lipids 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 heating and drying method.

[0058] The beer-flavored beverage of the present invention typically contains peptides with a molecular weight of 35 to 50 kDa derived from malt or the like. Peptides with a molecular weight of 35 to 50 kDa are peptides found in the 35 to 50 kDa range when measured by electrophoresis using SDS-PAGE after ultrafiltration of the raw material liquid of the beer-flavored beverage using a 30 kDa cutoff membrane. Preferably, the peptides are about 40 kDa, and in this specification, peptides of about 40 kDa are also referred to as 40 kDa peptides. In the beer-flavored beverage of the present invention, the content of 40kDa peptide is not particularly limited, but is preferably 5 to 60 ppm, and more preferably 10 to 40 ppm. In this specification, the 40 kDa peptide content refers to the value measured by the Bradford method.

[0059] The beer-flavored beverage of the present invention contains other peptides derived from malt, etc., in addition to the 40kDa peptide described above. These other peptides are peptides with a molecular weight of less than 35kDa or greater than 50kDa. More specifically, they refer to peptides other than the 40kDa peptide found in the molecular weight range of 35 to 50kDa (the 40kDa peptide) when the raw material liquid of the beer-flavored beverage is ultrafiltered using a 30kDa cutoff membrane and then measured by electrophoresis using SDS-PAGE. In the beer-flavored beverage of the present invention, the total peptide content, which is the sum of the 40kDa peptide and other peptides, is not particularly limited, but is preferably 60ppm or more, 80ppm or more, 100ppm or more, 200ppm or more, 400ppm or more, 600ppm or more, 800ppm or more, 900ppm or more, 1000ppm or more, 1100ppm or more, or 1200ppm or more, and preferably 4000ppm or less, 3800ppm or less, 3600ppm or less, 3400ppm or less, 3200ppm or less, or 3000ppm or less. In this specification, total peptide content refers to the value measured by the Lowry method.

[0060] In the beer-flavored beverage of the present invention, the total polyphenol content is not particularly limited, but from the viewpoint of taste, it may be, for example, 40 ppm or more, 42 ppm or more, 44 ppm or more, 46 ppm or more, 48 ppm or more, 50 ppm or more, 51 ppm or more, 52 ppm or more, 53 ppm or more, 54 ppm or more, 55 ppm or more, 56 ppm or more, 57 ppm or more, 58 ppm or more, 59 ppm or more, 60 ppm or more, or 400 ppm or less, 350 ppm or less, 300 ppm or less, 280 ppm or less, 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, 120 ppm or less, 110 ppm or less, or 100 ppm or less.

[0061] In this specification, total polyphenol content refers to the total amount of polyphenols contained in beer-flavored beverages. Polyphenols are not particularly limited, but examples include xanthohumol, catechin, hesperidin, chlorogenic acid, resveratrol, quercetin, anthocyanogen, and tannin. The total polyphenol content in beer-flavored beverages can be measured, for example, by the method described in section 8.19, Total Polyphenols, of the Revised BCOJ Beer Analysis Method (published by the Japan Brewing Association, edited by the International Technical Committee [Analysis Committee] of the Beer Brewers Association, 2013 revised and augmented edition).

[0062] The adjustment of the total polyphenol content involves the addition of dilution water or carbonated water, the type and amount of raw materials (polyphenol-containing raw materials such as malt, corn grits, and sugar solution), 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 (from malt addition to the wort production process before yeast addition), the wort filtration time, the set temperature and holding time for each temperature range when preparing the wort (including during saccharification), and the pre-fermentation liquid. The process can be carried out by appropriately setting the original extract concentration, the original extract concentration during the fermentation process, fermentation conditions (oxygen concentration, aeration conditions, yeast variety, amount of yeast added, number of yeast cells, timing of yeast removal, fermentation temperature, fermentation time, pressure setting, carbon dioxide concentration, etc.), cooling timing, cooling temperature, cooling time, beer filtration method (diatomaceous earth, membrane, sheet, cartridge, filter, etc.), activated carbon, and stabilizers added during beer filtration (silica gel, PVPP (polyvinyl polypyrrolidone), bentonite, tannin, etc.).

[0063] Furthermore, the total polyphenol content of the beer-flavored beverage of the present invention can be controlled by adjusting the amount of raw materials with high polyphenol content, such as barley malt and malt husks. 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, corn products (corn grits, corn protein, etc.), wheat, and wheat malt have a high nitrogen content but low polyphenol content. Therefore, the total nitrogen and total polyphenol content in beer-flavored beverages can be increased or decreased by adjusting the blending ratio of the raw materials. Below are some representative methods (1) to (4) for increasing or decreasing the total nitrogen and total polyphenol content. (1) Increase the total nitrogen content and total polyphenol content of beer-flavored beverages by increasing the amount of malt containing husks. (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. used, the total polyphenol content is 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 amount of nitrogen is maintained while reducing the total polyphenol content.

[0064] In the beer-flavored beverage of the present invention, from the viewpoint of providing a beverage with a more desirable depth of flavor, the total nitrogen content is, for example, 10 mg / 100 mL or more, 15 mg / 100 mL or more, 18 mg / 100 mL or more, 20 mg / 100 mL or more, 22 mg / 100 mL or more, 24 mg / 100 mL or more, 26 mg / 100 mL or more, 28 mg / 100 mL or more, 30 mg / 100 mL or more, 32 mg / 100 mL or more, 34 mg / 100 mL or more, 36 mg / 100 mL or more, 38 mg / 100 It may be 100mg / 100mL or more, or 40mg / 100mL or more, and from the viewpoint of making a beverage that is easy to swallow, it may be 100mg / 100mL or less, 90mg / 100mL or less, 80mg / 100mL or less, 70mg / 100mL or less, 60mg / 100mL or less, 55mg / 100mL or less, 50mg / 100mL or less, 48mg / 100mL or less, 46mg / 100mL or less, 44mg / 100mL or less, 42mg / 100mL or less, or 40mg / 100mL or less. In this specification, total nitrogen is the sum of all nitrogen compounds, including proteins and compounds containing amino acids. The total nitrogen content of the beer-flavored beverage of the present invention can be measured, for example, by the method described in "8.9 Total Nitrogen" of the Revised BCOJ Beer Analysis Method (published by the Japan Brewing Association, edited by the International Technical Committee [Analysis Committee] of the Beer Brewers Association, 2013 revised and augmented edition).

[0065] The total nitrogen content can be adjusted by appropriately setting the beer filtration conditions, such as the addition of dilution water or carbonated water, the type and amount of raw materials (malt, corn grits, sugar solution, etc.), the type of enzyme, the amount of enzyme (including proteolytic enzymes, etc.) added, the temperature during the enzymatic reaction, the timing of enzyme addition, the proteolytic time in the mashing tank, the pH in the mashing tank, the temperature in the mashing tank, the pH during the mashing process (the wort production process from malt addition to before yeast addition), the temperature during the mashing process, the temperature during wort filtration, the time during wort filtration, the pH during wort filtration, the amount of sparging water used during wort filtration, the set temperature and holding time for each temperature range when preparing the wort, the boiling time and pH during the boiling process, the original extract concentration of the pre-fermentation liquid, the original extract concentration during 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.).

[0066] The beer-flavored beverage of the present invention may be colorless and transparent, or it may be colored. In this specification, the "chromaticity" of the beverage can be measured by the measurement method described in "8.8 Chromaticity" of the Revised BCOJ Beer Analysis Method (published by the Japan Brewing Association, edited by the International Technical Committee [Analysis Committee] of the Beer Brewers Association, 2013 revised and augmented edition). The "chromaticity" of the beverage is specified by the unit of chromaticity (EBC unit) defined by the European Brewery Convention. A smaller value indicates a lighter and brighter colored beverage, while a larger value indicates a darker and more intense colored beverage. The color can be 0 EBC, or 1 EBC or higher, 5 EBC or higher, 10 EBC or higher, 15 EBC or higher, 20 EBC or higher, 30 EBC or higher, or 40 EBC or higher. It can also be 200 EBC or lower, 150 EBC or lower, 100 EBC or lower, 50 EBC or lower, 30 EBC or lower, 20 EBC or lower, 15 EBC or lower, or 10 EBC or lower. Color can be adjusted by appropriately setting the following: the addition of dilution water or carbonated water, the type and amount of raw materials (malt, corn grits, sugar solution, etc.), the temperature during mashing, the pH during mashing, the mashing time, the wort filtration time, the pH of the wort filtration, the boiling time, the boiling temperature, the amount of coloring components such as caramel coloring, the type of beer filtration (diatomaceous earth filtration, various membrane filtrations, etc.), and the amount of beer filtration. The color of the beer-flavored beverage of the present invention can be controlled by appropriately adjusting, for example, the type of malt used, the blending ratio if two or more types of malt are used in combination, the ratio of malt to other ingredients, and the boiling conditions when preparing the pre-fermentation liquid. More specifically, for example, to increase the color of the beer-flavored beverage, it can be adjusted by increasing the blending ratio of dark malt, increasing the boiling temperature, increasing the boiling time, and performing decoction when preparing the saccharified liquid. The color can also be adjusted to be higher by increasing the concentration of the raw wort extract or increasing the malt ratio. It can also be adjusted by controlling the amount of food additives such as caramel coloring or colored sugar solution.

[0067] The beer-flavored beverage of the present invention may be a packaged beverage. Any form and material of container may be used for the packaged beverage. Examples of containers include bottles, cans, kegs, or PET bottles, but cans, bottles, and PET bottles are particularly preferred from the viewpoint of ease of carrying.

[0068] <<Manufacturing method for beer-flavored beverages>> Next, the method for producing the beer-flavored beverage of the present invention (hereinafter also referred to simply as the "production method of the present invention" in this specification) will be described in detail. The present invention provides a method for producing a beer-flavored beverage with a malt ratio of 50% by weight or more, which includes a purine content adjustment step that adjusts the difference between the purine content (a) measured by method A below and the purine content (b) measured by method B below to 4.0 ppm or less. Method A: A method in which a 70% by weight perchloric acid aqueous solution is added to a beer-flavored beverage to a concentration of 18.2 v / v%, treated at 98°C for 1 hour, neutralized by adding a 4M potassium hydroxide aqueous solution to a concentration of 41 v / v%, and the purine base content in the obtained sample is measured by liquid chromatography-tandem mass spectrometry (LC-MS / MS). Method B: A method for measuring the purine base and nucleoside content in beer-flavored beverages by liquid chromatography-tandem mass spectrometry (LC-MS / MS) and calculating the sum of the purine base content and the nucleoside content in terms of purine base. According to the manufacturing method of the present invention, it is possible to provide a beer-flavored beverage with a reduced content of high-molecular-weight purines.

[0069] <Purine content adjustment process> The purine content adjustment step is a step of adjusting the difference between the purine content (a) measured by method A above and the purine content (b) measured by method B above to 4.0 ppm or less. The purine content adjustment step is also a step of adjusting the purine base content of high molecular weight purines to 4.0 ppm or less. In the manufacturing method of the present invention, the content of high molecular weight purines in a beer-flavored beverage can be reduced by performing a purine content adjustment step at any time between the brewing step and the filling step. Specific methods for reducing the content of high molecular weight purines in the purine content adjustment step include, for example, an adsorption step in which high molecular weight purines contained in the wort or wort fermentation liquid are adsorbed onto an adsorbent such as a cation exchange resin or activated carbon. These methods may be performed individually or in combination of two or more methods. The purine content adjustment step preferably includes an adsorption step in which purines contained in the wort or wort ferment liquor are adsorbed onto a cation exchange resin, and preferably includes an adsorption step in which high molecular weight purines contained in the wort or wort ferment liquor are adsorbed onto a cation exchange resin.

[0070] In this specification, "wort" refers to the liquid from which the components of malt have been dissolved, prior to fermentation by brewing yeast. "Wort" is a concept that includes the saccharified liquid obtained in the saccharification process. Furthermore, in this specification, "wort ferment liquor" refers to the liquid from which the components of malt have been dissolved, after fermentation by brewing yeast. "Wort ferment liquor" is a concept that includes fermented beer-flavored beverages. In this specification, "wort" and "wort ferment liquor" are clearly distinguished.

[0071] In this specification, "ion exchange resin" refers to a synthetic resin in which functional groups (ion exchange groups) are introduced into a polymer having a three-dimensional network structure. It separates various ions contained in a solution by utilizing the difference in adsorption between fixed ions in the ion exchange resin and alleles in various solutions. Styrene-based and acrylic-based polymer bases are common for ion exchange resins, and they are broadly classified into cation exchange resins, which exhibit acidic functional groups, and anion exchange resins, which exhibit basic functional groups. Furthermore, depending on the type of ion exchange group introduced, they can be classified into strongly acidic cation exchange resins, weakly acidic cation exchange resins, strongly basic anion exchange resins, weakly basic anion exchange resins, chelate resins that selectively adsorb heavy metal ions, etc.

[0072] In the manufacturing method of the present invention, if the purine content adjustment step includes an adsorption step in which purines contained in the wort or wort ferment liquor are adsorbed onto a cation exchange resin, the cation exchange resin is not particularly limited as long as its functional groups are acidic and capable of adsorbing cations in the solvent, and strongly acidic cation exchange resins and weakly acidic cation exchange resins can be suitably used. Examples of such cation exchange resins include AMBERLITE 200CT H AG (200CTH) (strong acid cation exchange resin), AMBERLITE IR120B H AG (strong acid cation exchange resin), AMBERLITE IR 124H H AG (strong acid cation exchange resin), AMBERLITE 252NA (strong acid cation exchange resin), AMBERLITE IRC76AG (weak acid cation exchange resin), and AMBERLITE FPC3500 (weak acid cation exchange resin) (all manufactured by Organo Corporation), DIAION PK216 (strong acid cation exchange resin), DIAION SK1BH (strong acid cation exchange resin), DIAION SK104 (strong acid cation exchange resin), DIAION SK1B (strong acid cation exchange resin), DIAION SK110 (strong acid cation exchange resin), DIAION UBK08, and DIAION Examples include, but are not limited to, UBK10, DIAION UBK12, and Relite® WK60L (all manufactured by Mitsubishi Chemical Corporation; DIAION is a registered trademark of Mitsubishi Chemical Corporation).

[0073] The method for adsorbing purines contained in wort or wort ferment liquor onto a cation exchange resin is not particularly limited as long as the wort or wort ferment liquor and the cation exchange resin can be brought into contact. For example, the wort or wort ferment liquor can be brought into contact with the cation exchange resin by mixing the cation exchange resin with the wort or wort ferment liquor. In this case, after mixing the cation exchange resin with the wort or wort ferment liquor, the mixture can be allowed to stand to precipitate the cation exchange resin that has adsorbed the purines, and the supernatant can be collected, or the cation exchange resin that has adsorbed the purines can be separated by centrifugation, thereby reducing the purine content in the wort or wort ferment liquor. Alternatively, the wort or wort ferment liquor may be brought into contact with the cation exchange resin by packing the cation exchange resin into a column and passing the wort or wort ferment liquor through the column. The cation exchange resin is not particularly limited, and conventionally known cation exchange resins can be used.

[0074] In the adsorption process, the amount of cation exchange resin used relative to the wort or wort ferment is not particularly limited, but is preferably 500 to 4000 ppm, more preferably 700 to 3500 ppm, and even more preferably 800 to 2500 ppm.

[0075] In the adsorption process, when purines contained in the wort ferment liquor are adsorbed onto a cation exchange resin or activated carbon, the ratio of the xanthine content to the total adenine and guanine content (xanthine content / total adenine and guanine content) of the wort ferment liquor is preferably less than 0.5, more preferably 0.4 or less, and even more preferably 0.3 or less.

[0076] In the manufacturing method of the present invention, the purine content adjustment step may further include a step of decomposing xanthine contained in the wort or wort ferment liquor. In this specification, decomposing xanthine means decomposing xanthine to at least uric acid, and includes assimilation of xanthine. The step of decomposing xanthine is not particularly limited as long as it can decompose xanthine to at least uric acid. Specific examples of the step of decomposing xanthine include a step of using yeast having xanthine assimilation ability to assimilate xanthine (xanthine assimilation step), a step of decomposing xanthine using xanthine oxidase (xanthine oxidase treatment step), and so on. These steps may be performed individually or in combination. Furthermore, when performed in combination, the order is not particularly limited, and the xanthine assimilation step and the xanthine oxidase treatment step may be performed simultaneously, the xanthine oxidase treatment step may be performed after the xanthine assimilation step, or the xanthine assimilation step may be performed after the xanthine oxidase treatment step.

[0077] In the xanthine assimilation process, yeast possessing xanthine assimilation ability is preferably yeast that has the XAN2 gene. The XAN2 gene is a gene that encodes 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. Yeast possessing the XAN2 gene is able to assimilate xanthine through the action of xanthine oxidase.

[0078] Examples of yeasts that possess xanthine assimilation ability include the genera Torulaspora, Lachancea, Zygosaccharomyces, Kluyveromyces, Hanseniaspora, Barnettozyma, Cyberlindnera, Wickerhamomyces, Ascoidea, Saccharomycopsis, Babjeviella, Candida / Lodderomyces, Debaryomyces, Diutina, Hyphopichia, Meyerozyma, Millerozyma, and Sch. Examples of yeasts belonging to the genera ffersomyces, Spathaspora, Suhomyces, Yamadazyma, Clavispora, Metschnikowia, Citeromyces, Komagataella, Kuraishia, Ambrosiozyma, Brettanomyces, Ogataea, Pichia, Saturnispora, Nakazawaea, Pachysolen, Peterozyma, Nadsonia, Galactomyces, Saprochaete, Yarrowia, Sugiyamaella, Tortispora, or Lipomyces include yeasts belonging to these genera. All yeasts belonging to these genera possess the XAN2 gene. Yeast with xanthine assimilation ability may be used individually or in combination of two or more types.

[0079] Examples of yeasts belonging to the genus Torulaspora include Torulaspora delbrueckii and Torulaspora pretoriensis.

[0080] Examples of yeasts belonging to the genus Lachancea include Lachancea fermentati and Lachancea thermotolerans.

[0081] Examples of yeasts belonging to the genus Zygosaccharomyces include Zygosaccharomyces rouxii.

[0082] Examples of yeasts belonging to the genus Kluyveromyces include Kluyveromyces marxianus.

[0083] In the purine content adjustment step, the yeast having xanthine assimilation ability is preferably at least one 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, specifically Torulaspora delbrueckii, Torulaspora pretoriensis, Lachancea fermentati, Lachancea thermotolerans, Zygosaccharomyces rouxii, and Kluyveromyces marxianus. It is more preferable that it be at least one species selected from the group consisting of (marxianus), and even more preferable that it be at least one species selected from the group consisting of Torulaspora delbrueckii, Lachancea fermentati, Zygosaccharomyces rouxii, and Kluyveromyces marxianus, and particularly preferable that it be Torulaspora delbrueckii.

[0084] As the yeast possessing xanthine assimilation ability, commercially available products may be used, or yeast isolated and purified from nature may be used. Alternatively, these yeasts possessing xanthine assimilation ability may be grown in advance by pre-culture, and the resulting pre-culture solution may be used in the xanthine assimilation process. Furthermore, if the yeast is dried, it may be rehydrated by conventionally known methods before being used in the xanthine assimilation process.

[0085] The culture conditions for pre-culturing yeast with xanthine assimilation ability are not particularly limited as long as the yeast with xanthine assimilation ability can grow, and can be set appropriately depending on the type of yeast with xanthine assimilation ability. The culture temperature may be, for example, 10 to 35°C, and is preferably 20 to 30°C. The culture time may be, for example, 5 hours to 5 days, and is preferably 16 hours to 3 days.

[0086] In the xanthine assimilation process, a method for causing yeast capable of assimilating xanthine to assimilate xanthine is, for example, to add yeast capable of assimilating xanthine to the wort or wort ferment, and then carry out fermentation by the yeast in the wort or wort ferment.

[0087] In the xanthine assimilation process, when yeast capable of assimilating xanthine is added to the wort or wort ferment, it is preferable that the wort or wort ferment contains xanthine. The xanthine content in the wort or wort ferment is not particularly limited, but may be, for example, 0.1 to 50 ppm, and preferably 5 to 30 ppm.

[0088] In the xanthine assimilation process, when adding yeast capable of xanthine assimilation to the wort or wort ferment, the amount of yeast capable of xanthine assimilation added is not particularly limited, but for example, 1 to 200 × 10⁻¹⁰ of the wort or wort ferment. 6 The number of cells / mL may be 10-100 × 10 6 cells / mL is preferred.

[0089] The fermentation conditions for using xanthine-assimilating yeast are not particularly limited as long as the xanthine-assimilating yeast can assimilate xanthine, and can be appropriately set depending on the type of xanthine-assimilating yeast, the amount added, the xanthine content in the wort or wort ferment. 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.

[0090] In the purine content adjustment process, the xanthine assimilation process can be carried out at any time during the manufacturing process of beer-flavored beverages, from the saccharification process to the filling process. For example, yeast with xanthine assimilation ability may be added to the wort at the same time as the regular brewing yeast and fermented (primary fermentation), or yeast with xanthine assimilation ability may be added to the wort ferment obtained by fermentation using regular brewing yeast and fermented further (secondary fermentation).

[0091] In the manufacturing method of the present invention, it is preferable that the purine content adjustment step further includes a xanthine assimilation step in which xanthine contained in the wort or wort ferment liquor is assimilated by yeast having xanthine assimilation ability. When the purine content adjustment step further includes a xanthine assimilation step, the xanthine content contained in the wort or wort ferment liquor can be reduced, and therefore the purine content (a) contained in the beer-flavored beverage can be reduced. In the manufacturing method of the present invention, it is even more preferable that the purine content adjustment step further includes a xanthine assimilation step before the adsorption step in which xanthine contained in the wort or wort ferment liquor is assimilated by yeast having xanthine assimilation ability.

[0092] In the xanthine oxidase treatment process, commercially available xanthine oxidase may be used, or xanthine oxidase isolated and purified from nature may be used. Alternatively, xanthine oxidase derived from yeast having xanthine assimilation ability may be used. Examples of xanthine oxidase derived from yeast having xanthine assimilation ability include: a crude enzyme solution obtained by disrupting yeast or its pre-culture using a conventionally known cell disruption method; an extract obtained by further extracting the contents of yeast having xanthine assimilation ability from the crude enzyme solution into an extract (bacterial cell extract); and a solution obtained by removing bacterial cell residue from the crude enzyme solution or bacterial cell extract. In the purine content adjustment process, when a xanthine oxidase treatment step is performed as a step to decompose xanthine, it is preferable to use xanthine oxidase derived from yeast that has xanthine assimilation ability.

[0093] In the xanthine oxidase treatment process, one method for decomposing xanthine using xanthine oxidase is, for example, adding xanthine oxidase to the wort or wort ferment liquor. The conditions in the xanthine oxidase treatment process, 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.

[0094] In the xanthine oxidase treatment process, when xanthine oxidase is added to the wort or wort ferment, it is preferable that the wort or wort ferment contains xanthine. The xanthine content in the wort or wort ferment is not particularly limited, but may be, for example, 0.1 to 50 ppm, and preferably 5 to 30 ppm.

[0095] In the xanthine oxidase treatment process, when xanthine oxidase is added to the wort or wort ferment, the amount added is not particularly limited, but it is preferable that the amount is such that the activity value of the 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 refers to the relative enzyme amount, where 1 unit (U) is defined as the amount of enzyme that produces 1 μmol of uric acid per minute under the following conditions. Mix 2.9 ml of 50 mM Tris-HCl buffer and 0.1 ml of 10 mM xanthine aqueous solution, preheat to 37°C, then add 0.01 ml of the solution of the substance to be measured, gently mix, and measure the change in absorbance at 293 nm per minute using a spectrophotometer controlled at 37°C with water as a control. From this value, the enzyme amount, where 1 μmol of enzyme per minute produces uric acid is defined as 1 unit (U), is taken as the activity value of xanthine oxidase.

[0096] In the purine content adjustment process, the xanthine oxidase treatment process can be carried out at any time during the beer-flavored beverage manufacturing process, from the saccharification process to the filling process. For example, xanthine oxidase may be added to the wort simultaneously with the usual brewing yeast to break down xanthine, or xanthine oxidase may be added to the wort ferment liquor obtained by fermentation using the usual brewing yeast to break down xanthine.

[0097] In the purine content adjustment step, it is preferable to add yeast or xanthine oxidase having xanthine assimilation ability to the wort or wort ferment liquor in the step of decomposing xanthine.

[0098] In the purine content adjustment process, by performing a xanthine assimilation process or a xanthine oxidase treatment process as a process to decompose xanthine, the xanthine content in the beer-flavored beverage can be reduced, thereby reducing the purine content (a). In the purine content adjustment process, the step of decomposing xanthine is preferably a xanthine utilization step.

[0099] In the manufacturing method of the present invention, the purine content adjustment step may further include a nucleosidase treatment step in which the wort or wort ferment liquor is treated with nucleosidase. The nucleosidase treatment step can be performed at any time during the manufacturing process of the beer-flavored beverage, from the mashing step to the filling step, and may be performed before the xanthine decomposition step, simultaneously with the xanthine decomposition step, or after the xanthine decomposition step. However, it is preferable to perform it before the xanthine decomposition step, and more preferably before the xanthine assimilation step. When performing the nucleosidase treatment step, nucleosidase may be added as a raw material for the beer-flavored beverage in the mashing step, or nucleosidase may be added to the wort or wort ferment liquor at any time between the saccharification step and the filling step.

[0100] By performing the nucleosidase treatment process, four types of nucleosides (adenosine, guanosine, inosine, and xanthosine) among the purines are hydrolyzed by the action of nucleosidase, becoming purine bases (adenine, guanine, hypoxanthine, and xanthine), thus reducing the content of the four types of nucleosides (adenosine, guanosine, inosine, and xanthosine). On the other hand, the content of the purine bases (adenine, guanine, hypoxanthine, and xanthine) increases, but adenine, guanine, and hypoxanthine can be assimilated by common brewing yeast, and furthermore, xanthine can be broken down in the xanthine decomposition process described above (preferably the xanthine assimilation process). Therefore, by performing the nucleosidase treatment process before the xanthine decomposition process (preferably the xanthine assimilation process), the purine content (a) in beer-flavored beverages can be further reduced. In the manufacturing method of the present invention, it is more preferable that the purine content adjustment step further includes a nucleosidase treatment step in which the wort or wort ferment liquor is treated with nucleosidase before the xanthine assimilation step.

[0101] In the purine content adjustment process, when a nucleosidase treatment step is performed, the amount of nucleosidase added is not particularly limited, but it is preferable that the amount is such that the activity value 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.

[0102] In this specification, the activity value of a nucleosidase is defined by quantifying the amount of ribose produced by a reaction using guanosine as a substrate. Each 1 mL of reaction solution contains 0.1 M acetate buffer (pH 4.3), 8 mM guanosine, and an appropriate amount of enzyme. The reaction is initiated by adding guanosine and carried out at 55°C for 30 minutes. After stopping the reaction by adding 1.5 mL of 0.5% dinitrosalicylic acid solution, the solution is boiled for 10 minutes. The absorbance of the cooled reaction solution at 540 nm is measured, and the activity value is calculated by subtracting the absorbance of the reaction solution without enzyme. The amount of enzyme that produces 1 μmol of ribose in 30 minutes is defined as 1 U of enzyme activity.

[0103] In the purine content adjustment process, the adsorption process, xanthine decomposition process, and nucleosidase treatment process described above may be appropriately combined with other methods for reducing purine content. Methods for reducing purine content other than the adsorption process, xanthine decomposition process, and nucleosidase treatment process are not particularly limited, but include, for example, methods of keeping the malt ratio low, dilution methods, and methods of adsorbing and removing free purines using adsorbents such as activated clay and zeolite. These can be carried out at any time from the mashing process to the filling process. These methods may be carried out individually or in combination of two or more.

[0104] In the manufacturing method of the present invention, it is preferable to adjust the purine content (a) in the purine content adjustment step to 14.0 ppm or less, 13.0 ppm or less, 12.0 ppm or less, 11.0 ppm or less, 10.0 ppm or less, 9.0 ppm or less, 8.0 ppm or less, 7.0 ppm or less, 6.0 ppm or less, 5.0 ppm or less, or less than 5.0 ppm. In the purine content adjustment step, the lower limit of the purine content (a) is not particularly limited, but can be adjusted to, for example, 0 ppm, 0.01 ppm, 0.02 ppm, 0.03 ppm, 0.04 ppm, 0.05 ppm, 0.06 ppm, 0.07 ppm, 0.08 ppm, 0.09 ppm, or 0.1 ppm.

[0105] In the manufacturing method of the present invention, except for the purine content reduction step described above, the raw materials and manufacturing process for the beer-flavored beverage, the content of each component in the resulting beer-flavored beverage, and preferred embodiments thereof are the same as those of the beer-flavored beverage of the present invention described above.

[0106] <<Methods for reducing purines in beer-flavored beverages>> Next, the method for reducing purines in beer-flavored beverages according to the present invention (hereinafter also referred to simply as the purine reduction method of the present invention in this specification) will be described in detail. The present invention provides a method for reducing purines in a beer-flavored beverage having a malt ratio of 50% by weight or more, which includes a purine content adjustment step of adjusting the difference between the purine content (a) measured by method A below and the purine content (b) measured by method B below to 4.0 ppm or less. Method A: A method in which a 70% by weight perchloric acid aqueous solution is added to a beer-flavored beverage to a concentration of 18.2 v / v%, treated at 98°C for 1 hour, neutralized by adding a 4M potassium hydroxide aqueous solution to a concentration of 41 v / v%, and the purine base content in the obtained sample is measured by liquid chromatography-tandem mass spectrometry (LC-MS / MS). Method B: A method for measuring the purine base and nucleoside content in beer-flavored beverages by liquid chromatography-tandem mass spectrometry (LC-MS / MS) and calculating the sum of the purine base content and the nucleoside content in terms of purine base. According to the purine reduction method of the present invention, it is possible to reduce the content of high-molecular-weight purines in beer-flavored beverages.

[0107] In the purine reduction method of the present invention, the raw materials and manufacturing process (including the purine content reduction process) of the beer-flavored beverage, the content of each component in the resulting beer-flavored beverage, and preferred embodiments thereof are the same as in the manufacturing method of the present invention described above.

[0108] In this specification, a numerical range expressed by a lower limit and an upper limit, i.e., "lower limit to upper limit," includes those lower and upper limits. For example, a range expressed as "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 any combination of ranges. [Examples]

[0109] The present invention will be described in more detail below with reference to examples, but this will not limit the scope of the present invention.

[0110] (Materials used) Commercially available beer-flavored beverage A (malt ratio: 50% by weight or more, real extract concentration: 0.93% by weight, carbohydrate content: 0.3g / 100mL, alcohol content: 5.5v / v%) Cation exchange resin A: Sulfonic acid-based cation exchange resin (Product name: AMBERLITE 200CT H AG (200CTH), manufactured by Organo Corporation) Cation exchange resin B: Sulfonic acid-based cation exchange resin (Product name: DIAION® PK216, manufactured by Mitsubishi Chemical Corporation) Cation exchange resin C: Sulfonic acid-based cation exchange resin (Product name: DIAION® SK1BH, manufactured by Mitsubishi Chemical Corporation) Cation exchange resin D: Acrylic acid-based cation exchange resin (Product name: Relite® WK60L, manufactured by Mitsubishi Chemical Corporation)

[0111] <Comparative Example 1> (Nucleosidase treatment process) A base beer was prepared by adding 0.5 g / L of nucleosidase (product name: PNF-L, manufactured by Shin Nippon Chemical Co., Ltd., unit count: 1600 U / g) and 0.5% by weight of glucose to a commercially available beer-flavored beverage A. The activity level of nucleosidase in the base beer was 0.8 U.

[0112] (Xanthine utilization process) In this process, Torulaspora delbrueckii (LEVEL2 BIODIVA, LALLEMAND), a yeast capable of xanthine assimilation, was used for the xanthine assimilation reaction.

[0113] The wort used for the pre-culture of the above yeast was prepared by the following method. 50g of crushed barley malt was added to a mashing tank containing 200mL of warm water maintained at 45°C and held for 30 minutes. Then, the temperature was increased by 1°C per minute to 67°C and held for 50 minutes. After further increasing the temperature to 78°C, the mixture was filtered to remove the spent grain and obtain a saccharified liquid. The obtained saccharified liquid was boiled and filtered to obtain wort. 10 mL of wort was placed in a test tube, and Torulaspora delbrueckii was inoculated using one platinum loop for pre-culture. The resulting pre-culture of Torulaspora delbrueckii was used in the following reaction.

[0114] 250 mL of the above base beer was placed in a 1000 mL Erlenmeyer flask, and the pre-culture solution of Torulaspora delbrueckii was added to carry out the reaction. After the reaction, the reaction solution was filtered to obtain xanthine-assimilated beer-flavored beverage A. The obtained xanthine-assimilated beer-flavored beverage A was used as the sample for Comparative Example 1.

[0115] In the sample of Comparative Example 1, the ratio of xanthine content to the total content of adenine and guanine (xanthine content / total content of adenine and guanine) was 0.039. The adenine, guanine, and xanthine content was measured by Method A under the conditions of liquid chromatography-tandem mass spectrometry (LC-MS / MS) described later.

[0116] <Example 1> (Adsorption process using cation exchange resin) 20 mL of xanthine assimilation-treated beer-flavored beverage A obtained in Comparative Example 1 was placed in a 50 mL glass beaker, and 0.02 g of cation exchange resin A was added to obtain a mixture containing 1000 ppm of cation exchange resin A. The obtained mixture was stirred at 4°C and 800 rpm for 1 hour to adsorb the purines contained in the xanthine assimilation-treated beer-flavored beverage A obtained in Comparative Example 1 onto the cation exchange resin. The mixture was then allowed to stand at 4°C for 0.5 hours to precipitate the cation exchange resin A that had adsorbed the purines, and 15 mL of the supernatant was collected to be used as the sample for Example 1.

[0117] <Example 2> Samples for Example 2 were obtained in the same manner as in Example 1, except that cation exchange resin B was used instead of cation exchange resin A.

[0118] <Example 3> Sample for Example 3 was obtained in the same manner as in Example 1, except that cation exchange resin C was used instead of cation exchange resin A.

[0119] <Example 4> Sample for Example 4 was obtained in the same manner as in Example 1, except that cation exchange resin D was used instead of cation exchange resin A.

[0120] (Measurement of high molecular weight purine content) For the samples of Examples 1-4 and Comparative Example 1, the purine content (a) was measured using Method A below, and the purine content (b) was measured using Method B below. Method A: A method in which a 70% by weight perchloric acid aqueous solution is added to beer-flavored beverages (samples from Examples 1-4 and Comparative Example 1) to a concentration of 18.2 v / v%, treated at 98°C for 1 hour, neutralized by adding a 4M potassium hydroxide aqueous solution to a concentration of 41 v / v%, and the purine base content in the obtained sample is measured by liquid chromatography-tandem mass spectrometry (LC-MS / MS). Method B: A method for measuring the purine base and nucleoside content in beer-flavored beverages (samples from Examples 1-4 and Comparative Example 1) by liquid chromatography-tandem mass spectrometry (LC-MS / MS), and calculating the sum of the purine base content and the nucleoside content in terms of purine base.

[0121] In method A described above, the supernatant was collected by centrifugation using a tabletop centrifuge (product name: Tabletop Centrifuge 4000, manufactured by Kubota Shoji Co., Ltd.) at 800G for 5 minutes. Furthermore, in methods A and B described above, measurements by liquid chromatography-tandem mass spectrometry (LC-MS / MS) were performed using a liquid chromatograph-tandem mass spectrometer (LC-MS / MS) (SCIEX, X500R QTOF) under the following measurement conditions. Column: Discovery® HS F5 HPLC column (Product No.: 567503-U, 3μm particle size, L×ID 15cm×2.1mm, manufactured by Merck) Elution: Gradient using 0.1% formic acid / H2O (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℃

[0122] The purine base content of high molecular weight purines in the samples of Examples 1-4 and Comparative Example 1 was calculated based on the following formula. (Purine base content of high molecular weight purines) = (Purine content (a)) - (Purine content (b))

[0123] (Measurement of total polyphenol content) For each sample in Examples 1-4 and Comparative Example 1, the total polyphenol content was measured using the Forinthiocalto method (in terms of gallic acid) according to the following procedure. Each sample was diluted fivefold with distilled water to obtain a sample solution. To 0.50 mL of the sample solution, 2.5 mL of Forlinthiocaltophenol reagent (Sigma-Aldrich), diluted 10-fold with distilled water, was added, stirred, and reacted at room temperature for 10 minutes. Then, 2.0 mL of 0.795 mol / L sodium carbonate solution was added and mixed. After standing for 1 hour, the absorbance at 765 nm was measured using a spectrophotometer. A calibration curve was also created by measuring the absorbance of 0.50 mL of gallic acid standard solution in the same manner as the sample solution. Furthermore, a sample blank was performed in the same manner as above, except that 0.50 mL of water was used instead of 0.50 mL of the sample solution. The polyphenol concentration (A μg / mL) in the sample solution was determined from the calibration curve created by subtracting the absorbance of the sample blank from the absorbance of the sample solution.

[0124] (Measurement of amino acid content) The amino acid content of the samples from Examples 1-4 and Comparative Example 1 was measured by high-performance liquid chromatography (HPLC) according to the following procedure. ·Analytical equipment HPLC: Amino acid analysis system manufactured by Shimadzu Corporation. Autosampler: SIL-30AC • Pre-analysis processing Filter 10 mL of the sample through a membrane filter (pore size: 0.2 μm). ·Analysis conditions Column: Triart C18 (1.9 μm, manufactured by YMC Corporation) Mobile phase (gradient): (Solution A) 20 mmol / L potassium phosphate buffer (pH 6.9), (Solution B) acetonitrile / methanol / water (45 / 40 / 15) Flow rate: 0.8ml / min Temperature: 35℃ Detection: RF-20Axs (Ex 350nm, Em 450nm → Ex 255nm, Em 305nm (9.0 min)) Cell temperature: 20℃ Flow cell: Conventional cell Injection volume: 1μL

[0125] (Measurement of total peptide content) The total peptide content was measured using the Lowry method with a commercially available kit (DC Protein Assay, Bio-Rad). First, the samples were adjusted to appropriate concentrations (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). To 5 μL of the adjusted sample, 50 μL of solution A was added and mixed, followed by 400 μL of solution B and mixed. After a chromogenic reaction at room temperature for 15 minutes, 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 obtained absorbance and a pre-prepared calibration curve. The calibration curve was prepared using BSA (bovine serum albumin).

[0126] Table 1 shows the results of measuring the purine content (a), purine content (b), high molecular weight purine content (in purine base equivalent), total polyphenol content, amino acid content, and total peptide content for the samples of Examples 1-4 and Comparative Example 1.

[0127] [Table 1]

[0128] As shown in Table 1, in Examples 1 to 4, where an adsorption process using a cation exchange resin was performed, the content of high molecular weight purines in terms of purine bases was reduced by approximately 17 to 23% compared to Comparative Example 1, where no adsorption process using a cation exchange resin was performed. Furthermore, among Examples 1 to 4, Example 1, which used cation exchange resin A, showed the highest reduction rate in the content of high molecular weight purines in terms of purine bases. In addition, in Examples 1 to 4, the purine content (a) was reduced by 0.9 to 1.1 ppm compared to Comparative Example 1, and of that, the reduction in the content of high molecular weight purines in terms of purine bases accounted for 0.7 to 1.0 ppm. This indicates that it is possible to specifically adsorb high molecular weight purines (specifically reduce the content of high molecular weight purines) among purines through the adsorption process using a cation exchange resin. On the other hand, the total polyphenol content in Examples 1 to 4 was hardly reduced from the value in Comparative Example 1, and even in Example 2, which showed the highest reduction rate in total polyphenol content among Examples 1 to 4, the reduction rate was kept below 5%. Furthermore, in Examples 1 to 4, where an adsorption process using a cation exchange resin was performed, it was found that the purine content (b), the content and composition ratio of each amino acid, and the total peptide content remained almost unchanged compared to Comparative Example 1, where no adsorption process using a cation exchange resin was performed.

[0129] <Comparative Example 2> (Nucleosidase treatment process) A base beer was prepared by adding 0.5 g / L of nucleosidase (product name: PNF-L, manufactured by Shin Nippon Chemical Co., Ltd., unit count: 1600 U / g) and 0.5% by weight of glucose to a commercially available beer-flavored beverage A. The activity level of nucleosidase in the base beer was 0.8 U.

[0130] (Xanthine utilization process) In this process, Torulaspora delbrueckii (LEVEL2 BIODIVA, LALLEMAND), a yeast capable of xanthine assimilation, was used for the xanthine assimilation reaction.

[0131] The wort used for the pre-culture of the above yeast was prepared by the following method. 50g of crushed barley malt was added to a mashing tank containing 200mL of warm water maintained at 45°C and held for 30 minutes. Then, the temperature was increased by 1°C per minute to 67°C and held for 50 minutes. After further increasing the temperature to 78°C, the mixture was filtered to remove the spent grain and obtain a saccharified liquid. The obtained saccharified liquid was boiled and filtered to obtain wort. 10 mL of wort was placed in a test tube, and Torulaspora delbrueckii was inoculated using one platinum loop for pre-culture. The resulting pre-culture of Torulaspora delbrueckii was used in the following reaction.

[0132] 250 mL of the above base beer was placed in a 1000 mL Erlenmeyer flask, and the pre-culture solution of Torulaspora delbrueckii was added to carry out the reaction. After the reaction, the reaction solution was filtered to obtain xanthine-assimilated beer-flavored beverage A. The obtained xanthine-assimilated beer-flavored beverage A was used as the sample for Comparative Example 2.

[0133] In the sample of Comparative Example 2, the ratio of xanthine content to the total content of adenine and guanine (xanthine content / total content of adenine and guanine) was 0.263. The method for measuring the content of adenine, guanine, and xanthine is the same as the method used in Comparative Example 1.

[0134] <Example 5> (Adsorption process using cation exchange resin) 20 mL of xanthine assimilation-treated beer-flavored beverage A obtained in Comparative Example 2 was placed in a 50 mL glass beaker, and 0.02 g of cation exchange resin A was added to obtain a mixture containing 1000 ppm of cation exchange resin A. The obtained mixture was stirred at 4°C and 800 rpm for 1 hour to adsorb the purines contained in the xanthine assimilation-treated beer-flavored beverage A obtained in Comparative Example 2 onto the cation exchange resin. The mixture was then allowed to stand at 4°C for 0.5 hours to precipitate the cation exchange resin A that had adsorbed the purines, and 15 mL of the supernatant was collected to be used as the sample for Example 5.

[0135] <Example 6> Sample for Example 6 was obtained in the same manner as in Example 5, except that the amount of cation exchange resin A added was set to 0.1 g and the content of cation exchange resin A was changed to 5000 ppm.

[0136] (Measurement of high molecular weight purine content) For the samples of Examples 5 and 6 and Comparative Example 2, the purine content (a) was measured by Method A and the purine content (b) was measured by Method B, respectively, using the same method as in Example 1, and the purine base content of high molecular weight purines was calculated.

[0137] (Measurement of total polyphenol content) The total polyphenol content of the samples from Examples 5 and 6 and Comparative Example 2 was measured using the same method as in Example 1.

[0138] (Measurement of amino acid content) The amino acid content of the samples from Examples 5 and 6 and Comparative Example 2 was measured using the same method as in Example 1.

[0139] (Measurement of total peptide content) The total peptide content of the samples from Examples 5 and 6 and Comparative Example 2 was measured using the same method as in Example 1.

[0140] (Sensory evaluation) Four expert panelists conducted sensory evaluations of the samples from Examples 5 and 6 and Comparative Example 2 from the perspective of "taste." As a baseline, the evaluation of the Comparative Example 2 sample was set at 3.0 points, and the evaluation of the 5% ethanol aqueous solution was set at 0 points, with scores calculated in 0.5-point increments. A higher sensory evaluation score indicates a higher evaluation (better taste). Here, "good taste" means that the taste characteristic of a beer-flavored beverage is clearly discernible. The average score of the four expert panelists was used as the evaluation result.

[0141] Table 2 shows the measurement results for the purine content (a), purine content (b), high molecular weight purine content (in purine base equivalent), total polyphenol content, amino acid content, and total peptide content of the samples from Examples 5 and 6 and Comparative Example 2, as well as the results of the sensory evaluation.

[0142] [Table 2]

[0143] As shown in Table 2, in Examples 5 and 6, where an adsorption process using a cation exchange resin was performed, the content of high molecular weight purines in terms of purine bases was reduced by approximately 16-27% compared to Comparative Example 2, where no adsorption process using a cation exchange resin was performed. Furthermore, comparing the results of Examples 5 and 6, it was found that Example 6, which had a larger amount of cation exchange resin added, showed a higher reduction rate in the content of high molecular weight purines in terms of purine bases. In addition, in Examples 5 and 6, the purine content (a) was reduced by 0.7-1.3 ppm compared to Comparative Example 2, and of that, the reduction in the content of high molecular weight purines in terms of purine bases accounted for 0.7-1.2 ppm. This indicates that it is possible to specifically adsorb high molecular weight purines (specifically reduce the content of high molecular weight purines) among purines through the adsorption process using a cation exchange resin. On the other hand, the total polyphenol content in Example 5 was hardly reduced compared to Comparative Example 2, with a reduction rate of less than 1%, and in Example 6, the reduction rate compared to Comparative Example 2 was only about 5%. Furthermore, in Example 5, compared to Comparative Example 1, which did not undergo an adsorption process using a cation exchange resin, the purine content (b), the content and composition ratio of each amino acid, and the total peptide content remained almost unchanged, and the taste was not significantly impaired in the sensory evaluation and was within an acceptable range.

[0144] <Comparative Example 3> (Nucleosidase treatment process) A base beer was prepared by adding 0.5 g / L of nucleosidase (product name: PNF-L, manufactured by Shin Nippon Chemical Co., Ltd., unit count: 1600 U / g) and 0.5% by weight of glucose to a commercially available beer-flavored beverage A. The activity level of nucleosidase in the base beer was 0.8 U.

[0145] (Xanthine utilization process) In this process, Torulaspora delbrueckii (LEVEL2 BIODIVA, LALLEMAND), a yeast capable of xanthine assimilation, was used for the xanthine assimilation reaction.

[0146] The wort used for the pre-culture of the above yeast was prepared by the following method. 50g of crushed barley malt was added to a mashing tank containing 200mL of warm water maintained at 45°C and held for 30 minutes. Then, the temperature was increased by 1°C per minute to 67°C and held for 50 minutes. After further increasing the temperature to 78°C, the mixture was filtered to remove the spent grain and obtain a saccharified liquid. The obtained saccharified liquid was boiled and filtered to obtain wort. 10 mL of wort was placed in a test tube, and Torulaspora delbrueckii was inoculated using one platinum loop for pre-culture. The resulting pre-culture of Torulaspora delbrueckii was used in the following reaction.

[0147] 250 mL of the above base beer was placed in a 1000 mL Erlenmeyer flask, and the pre-culture solution of Torulaspora delbrueckii was added to carry out the reaction. After the reaction, the reaction solution was filtered to obtain xanthine-assimilated beer-flavored beverage A. The obtained xanthine-assimilated beer-flavored beverage A was used as the sample for Comparative Example 3.

[0148] In the sample of Comparative Example 3, the ratio of xanthine content to the total content of adenine and guanine (xanthine content / total content of adenine and guanine) was 0.066. The method for measuring the content of adenine, guanine, and xanthine is the same as the method used in Comparative Example 1.

[0149] <Example 7> (Adsorption process using cation exchange resin) 20 mL of xanthine assimilation-treated beer-flavored beverage A obtained in Comparative Example 3 was placed in a 50 mL glass beaker, and 0.02 g of cation exchange resin A was added to obtain a mixture containing 1000 ppm of cation exchange resin A. The obtained mixture was stirred at 4°C and 800 rpm for 1 hour to adsorb the purines contained in the xanthine assimilation-treated beer-flavored beverage A obtained in Comparative Example 3 onto the cation exchange resin. The mixture was then allowed to stand at 4°C for 0.5 hours to precipitate the cation exchange resin A that had adsorbed the purines, and 15 mL of the supernatant was collected to be used as the sample for Example 7.

[0150] <Example 8> Sample for Example 8 was obtained in the same manner as in Example 7, except that the amount of cation exchange resin A added was changed to 0.2 g and the content of cation exchange resin A was changed to 10,000 ppm.

[0151] (Measurement of high molecular weight purine content) For the samples of Examples 7 and 8 and Comparative Example 3, the purine content (a) was measured by Method A and the purine content (b) was measured by Method B, respectively, using the same method as in Example 1, and the purine base content of high molecular weight purines was calculated.

[0152] (Measurement of total polyphenol content) The total polyphenol content of the samples from Examples 7 and 8 and Comparative Example 3 was measured using the same method as in Example 1.

[0153] (Measurement of total peptide content) The total peptide content of the samples from Examples 7 and 8 and Comparative Example 3 was measured using the same method as in Example 1.

[0154] Table 3 shows the results of measuring the purine content (a), purine content (b), high molecular weight purine content (in purine base equivalent), total polyphenol content, and total peptide content for the samples of Examples 7 and 8 and Comparative Example 3.

[0155] [Table 3]

[0156] As shown in Table 3, in Examples 7 and 8, where an adsorption process using a cation exchange resin was performed, the content of high molecular weight purines in terms of purine bases was reduced by approximately 16-25% compared to Comparative Example 3, where no adsorption process using a cation exchange resin was performed. Furthermore, comparing the results of Examples 7 and 8, it was found that Example 8, which had a larger amount of cation exchange resin added, showed a higher reduction rate in the content of high molecular weight purines in terms of purine bases. On the other hand, the total polyphenol content in Examples 7 and 8 showed a reduction rate of approximately 2.1 to 6.3% compared to Comparative Example 3.

Claims

1. A beer-flavored beverage having a malt content of 50% by weight or more, and where the difference between the purine content measured by method A below (a) and the purine content measured by method B below (b) is 4.0 ppm or less. Method A: A method in which a 70% by weight perchloric acid aqueous solution is added to a beer-flavored beverage to a concentration of 18.2 v / v%, treated at 98°C for 1 hour, neutralized by adding a 4M potassium hydroxide aqueous solution to a concentration of 41 v / v%, and the purine base content in the obtained sample is measured by liquid chromatography-tandem mass spectrometry (LC-MS / MS). Method B: A method for measuring the purine base and nucleoside content in beer-flavored beverages by liquid chromatography-tandem mass spectrometry (LC-MS / MS) and calculating the sum of the purine base content and the nucleoside content in terms of purine base.

2. The beer-flavored beverage according to claim 1, wherein the purine content (a) is less than 5.0 ppm.

3. A beer-flavored beverage according to claim 1 or 2, wherein the real extract concentration is 0.5% by weight or more.

4. A beer-flavored beverage according to claim 1 or 2, wherein the total peptide content is 60 ppm or more.

5. A beer-flavored beverage according to claim 1 or 2, wherein the total polyphenol content is 50 ppm or more.

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

7. A method for producing a beer-flavored beverage with a malt ratio of 50% by weight or more, comprising a purine content adjustment step of adjusting the difference between the purine content (a) measured by method A below and the purine content (b) measured by method B below to 4.0 ppm or less. Method A: A method in which a 70% by weight perchloric acid aqueous solution is added to a beer-flavored beverage to a concentration of 18.2 v / v%, treated at 98°C for 1 hour, neutralized by adding a 4M potassium hydroxide aqueous solution to a concentration of 41 v / v%, and the purine base content in the obtained sample is measured by liquid chromatography-tandem mass spectrometry (LC-MS / MS). Method B: A method for measuring the purine base and nucleoside content in beer-flavored beverages by liquid chromatography-tandem mass spectrometry (LC-MS / MS) and calculating the sum of the purine base content and the nucleoside content in terms of purine base.

8. The method for producing a beer-flavored beverage according to claim 7, wherein the purine content adjustment step includes an adsorption step of adsorbing purines contained in the wort or wort fermentation liquid onto a cation exchange resin.

9. A method for producing a beer-flavored beverage according to claim 8, wherein the amount of cation exchange resin used relative to the wort or wort ferment liquor in the adsorption step is 500 to 4000 ppm.

10. A method for producing a beer-flavored beverage according to claim 8 or 9, wherein the purine content adjustment step further includes a xanthine assimilation step, prior to the adsorption step, in which xanthine contained in the wort or wort ferment liquor is assimilated by yeast having xanthine assimilation ability.

11. The method for producing a beer-flavored beverage according to claim 10, wherein the purine content adjustment step further comprises a nucleosidase treatment step of treating the wort or wort ferment with nucleosidase before the xanthine assimilation step.

12. The method for producing a beer-flavored beverage according to claim 8 or 9, wherein the wort fermentation liquid has a ratio of xanthine content to the total content of adenine and guanine (xanthine content / total content of adenine and guanine) of less than 0.

5.

13. A method for producing a beer-flavored beverage according to any one of claims 7 to 9, wherein in the purine content adjustment step, the purine content (a) is adjusted to less than 5.0 ppm.

14. A method for reducing purines in a beer-flavored beverage having a malt ratio of 50% by weight or more, comprising a purine content adjustment step of adjusting the difference between the purine content (a) measured by method A below and the purine content (b) measured by method B below to 4.0 ppm or less. Method A: A method in which a 70% by weight perchloric acid aqueous solution is added to a beer-flavored beverage to a concentration of 18.2 v / v%, treated at 98°C for 1 hour, neutralized by adding a 4M potassium hydroxide aqueous solution to a concentration of 41 v / v%, and the purine base content in the obtained sample is measured by liquid chromatography-tandem mass spectrometry (LC-MS / MS). Method B: A method for measuring the purine base and nucleoside content in beer-flavored beverages by liquid chromatography-tandem mass spectrometry (LC-MS / MS) and calculating the sum of the purine base content and the nucleoside content in terms of purine base.

15. The method for reducing the purine content of a beer-flavored beverage according to claim 14, wherein the purine content adjustment step includes an adsorption step of adsorbing purines contained in the wort or wort fermentation liquid onto a cation exchange resin.

16. A method for reducing purines in a beer-flavored beverage according to claim 15, wherein the amount of cation exchange resin used relative to the wort or wort ferment liquor in the adsorption step is 500 to 4000 ppm.

17. The method for reducing purines in a beer-flavored beverage according to claim 15 or 16, wherein the purine content adjustment step further includes a xanthine assimilation step, in which xanthine contained in the wort or wort ferment liquor is assimilated by yeast having xanthine assimilation ability, prior to the adsorption step.

18. The method for reducing purines in a beer-flavored beverage according to claim 17, wherein the purine content adjustment step further comprises a nucleosidase treatment step of treating the wort or wort ferment with nucleosidase prior to the xanthine assimilation step.

19. The method for reducing purines in a beer-flavored beverage according to claim 15 or 16, wherein the ratio of the xanthine content to the total adenine and guanine content (xanthine content / total adenine and guanine content) of the wort ferment is less than 0.

5.

20. A method for reducing the purine content of a beer-flavored beverage according to any one of claims 14 to 16, wherein in the purine content adjustment step, the purine content (a) is adjusted to less than 5.0 ppm.

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