Method for manufacturing beer-flavored fermented beverages

Abstract

Providing a novel manufacturing method for beer-flavored fermented beverages. [Solution] The present invention provides a method for producing a beer-flavored fermented beverage, comprising the step of fermenting using adenine-requiring yeast, wherein the adenine-requiring yeast is deficient in at least a part of the function of the enzyme that catalyzes the metabolism of the IMP biosynthesis pathway. The present invention is advantageous in that, in the production of a beer-flavored fermented beverage, fermentation using adenine-requiring yeast can be easily reduced in terms of the xanthine concentration of the beer-flavored fermented beverage.

Description

Technical Field

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

Background Art

[0002] It is said that about 4 to 10 mg / 100 mL (40,000 to 100,000 ppb) of purine compounds are present in beer, which is a fermented malt beverage. When purine compounds are ingested as food, they are decomposed into uric acid, which is said to contribute to an increase in uric acid levels. Therefore, for fermented malt beverages such as beer, products with reduced purine compounds that meet consumers' health orientation are desired.

[0003] So far, technologies for reducing the concentration of purine compounds in fermented malt beverages such as beer have been developed (for example, Patent Documents 1 and 2). Also, a beer-taste fermented alcoholic beverage with a reduced purine concentration and a realized richness in taste is known (Patent Document 3).

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Patent Document 2

Patent Document 3

Summary of the Invention

Problems to be Solved by the Invention

[0005] An object of the present invention is to provide a novel method for producing a beer-taste fermented beverage.

Means for Solving the Problems

[0006] The inventors have discovered that by using adenine-requiring yeast for fermentation in the production of beer-flavored fermented beverages, the amount of xanthine produced during fermentation can be reduced, thereby enabling the production of beer-flavored fermented beverages with reduced xanthine concentration. This invention is based on this finding.

[0007] The present invention provides the following inventions. [1] A method for producing a beer-flavored fermented beverage, comprising the step of fermenting using adenine-requiring yeast, wherein the adenine-requiring yeast is deficient in at least a part of the function of an enzyme that catalyzes the metabolism of the IMP biosynthesis pathway. [2] The manufacturing method according to [1] above, wherein the enzyme is one or more selected from the group consisting of ADE5,7, ADE4, ADE8, ADE6, ADE2, ADE1, ADE13, ADE16, ADE17, and ADE3. [3] The manufacturing method according to [1] above, wherein the enzyme is ADE5,7. [4] The manufacturing method according to any one of [1] to [3] above, wherein the malt usage ratio of the beverage is 50% or more and 90% or less. [5] The manufacturing method according to any one of [1] to [4] above, wherein the beverage has a xanthine concentration of 18,000 ppb or less and / or a purine concentration of 35,000 ppb or less. [6] The manufacturing method according to any one of [1] to [5] above, wherein the ratio of xanthine concentration (ppb) to purine concentration (ppb) of the beverage is 0.4 or less. [7] A beer-flavored fermented beverage having a xanthine concentration of 18,000 ppb or less and / or a purine concentration of 35,000 ppb or less, and containing adenine-requiring yeast. [8] The beverage described in [7] above, wherein the ratio of xanthine concentration (ppb) to purine concentration (ppb) is 0.4 or less. [9] The beverages described in [7] or [8] above, wherein the malt content is 50% or more and 90% or less.

[10] A method for reducing the xanthine concentration and / or purine concentration of a beer-flavored fermented beverage, comprising the step of fermenting using adenine-requiring yeast, wherein the adenine-requiring yeast is deficient in at least part of the function of an enzyme that catalyzes the metabolism of the IMP biosynthesis pathway.

[0008] According to the present invention, in the production of beer-flavored fermented beverages, it is advantageous that the xanthine concentration of the beer-flavored fermented beverage can be easily reduced by using adenine-requiring yeast for fermentation. [Brief explanation of the drawing]

[0009] [Figure 1] Figure 1 shows the IMP biosynthesis pathway (black arrow) in yeast (Saccharomyces pastorianus and Saccharomyces cerevisiae). In the figure, PRPP represents 5-phosphoribosyl-1α-bisphosphate, PRA represents 5-phospho-β-ribosylamine, GAR represents glycinamide lipotide, FGAR represents formylglycinamide lipotide, FGAM represents formylglycineamidine lipotide, AIR represents 5-aminoimidazole lipotide, CAIR represents 4-carboxy-5-aminoimidazole lipotide, SAICAR represents 5-aminoimidazole-4-(N-succinocarboxamide) lipotide, AICAR represents 5-aminoimidazole-4-carboxamide lipotide, FAICAR represents 5-formylaminoimidazole-4-carboxamide lipotide, and IMP represents inosinic acid. These are intermediates in purine biosynthesis. In the figure, each ADE represents the enzyme that catalyzes each metabolic step in the IMP biosynthesis pathway. In the diagram, GDP represents guanosine diphosphate, GMP represents guanylic acid, XMP represents xanthosine monophosphate, and AMP represents adenosine monophosphate. [Modes for carrying out the invention]

[0010] In this invention, "beer-flavored beverage" means a beverage having a beer-like flavor. In this invention, "beer-flavored fermented alcoholic beverage" means a beverage fermented with yeast using a carbon source, a nitrogen source, and water as raw materials, and includes beer, sparkling wine, and other sparkling alcoholic beverages (alcoholic beverages other than beer and sparkling wine that have an alcohol concentration of less than 11 v / v% and are effervescent). A preferred embodiment of the beer-flavored fermented alcoholic beverage is a beer-flavored fermented alcoholic beverage in which malt and / or ungerminated grains are used as at least part of the raw materials, and more preferably a beer-flavored fermented alcoholic beverage in which malt is used as at least part of the raw materials. In this invention, "beer-flavored fermented non-alcoholic beverage" means a beverage produced by removing the alcohol component produced in a beer-flavored fermented alcoholic beverage that has undergone a fermentation process using yeast, and a beverage produced by stopping the fermentation process at a stage where the ethanol concentration is less than 1 v / v%. In this invention, "beer-flavored fermented non-alcoholic beverage" includes beverages that contain no alcohol at all, i.e., beverages with an ethanol concentration of 0.00 v / v%, and beverages with an ethanol concentration greater than 0.00 v / v% and less than 1 v / v%. In this invention, "beer-flavored fermented beverage" includes both "beer-flavored fermented alcoholic beverage" and "beer-flavored fermented non-alcoholic beverage."

[0011] In this invention, "low-malt beer" includes those containing distilled alcohol as an ingredient, and those that do not contain malt or hops. In this invention, "beer" refers to a beverage made by fermenting malt, hops, and water, or a beverage made by fermenting malt, hops, water, and certain auxiliary ingredients, and that satisfies the following two conditions (limited to those with an alcohol content of less than 20%). • The malt ratio must be 50 / 100 or higher. • The weight of the fruit used (including dried, boiled, or concentrated fruit juice) and certain flavorings must not exceed 5% of the weight of the malt (including those not used). This can be interpreted as meaning (see the Liquor Tax Act (Law No. 6 of 1953), which came into effect on October 1, 2024).

[0012] The present invention relates to a method for producing a beer-flavored fermented beverage, characterized by including a step of fermentation using adenine-requiring yeast. In the present invention, "adenine-requiring yeast" refers to a type of brewing yeast that requires adenine for growth. The present invention is characterized by using adenine-requiring yeast in which at least a part of the function of an enzyme that catalyzes the metabolism of the IMP biosynthesis pathway is deficient. In the present invention, "IMP biosynthesis pathway (novel purine base synthesis pathway)" refers to the pathway in which inosinic acid (IMP) is synthesized from 5-phosphoribosyl-1α-bisphosphate (PRPP) in yeast, as shown in Figure 1 (Stepchenkova et al., BMC Genetics BMC Genet., 2005; 6: 31.). The adenine-requiring yeast used in the present invention is characterized by deficient in at least a part of the function of at least one enzyme that catalyzes the metabolism of the IMP biosynthesis pathway.

[0013] The yeast used in this invention is brewer's yeast, and the species is, for example, Saccharomyces pastorianus and Saccharomyces cerevisiae These are some examples.

[0014] In the present invention, enzymes that catalyze the metabolism of the IMP biosynthesis pathway include ADE5,7, ADE4, ADE8, ADE6, ADE2, ADE1, ADE13, ADE16, ADE17, and ADE3, which are encoded by the ADE gene. In the present invention, adenine-requiring yeast is characterized by a deficiency in the enzymatic activity of one or more ADEs selected from ADE5,7, ADE4, ADE8, ADE6, ADE2, ADE1, ADE13, ADE16, ADE17, and ADE3, preferably characterized by a deficiency in the function of ADE5,7, and more preferably characterized by a deficiency in at least the AIRS function of ADE5,7.

[0015] In the present invention, ADE5,7 (system name: YGL234W, SGD: S000003203) is a bifunctional enzyme having two functions of phosphoribosylglycinamidine synthetase (GARS) and phosphoribosylaminoimidazole synthetase (AIRS). ADE5,7 catalyzes the reaction from PRA to GAR and the reaction from FGAM to AIR in the IMP biosynthetic pathway. In this specification, the system name and SGD are based on the Saccharomyces Genome Database (SGD) (https: / / www.yeastgenome.org / ).

[0016] In the present invention, ADE4 (system name: YMR300C, SGD: S000004915) is phosphoribosylpyrophosphate amidotransferase and catalyzes the first reaction (the reaction from PRPP to PRA) in the IMP biosynthetic pathway.

[0017] In the present invention, ADE8 (system name: YDR408C, SGD: S000002816) is phosphoribosylglycinamide transformylase and catalyzes the reaction from GAR to FGAR in the IMP biosynthetic pathway.

[0018] In the present invention, ADE6 (system name: YGR061C, SGD: S000003293) is formylglycinamide nucleotide (FGAM) synthetase and catalyzes the reaction from FGAR to FGAM in the IMP biosynthetic pathway.

[0019] In the present invention, ADE2 (system name: YOR128C, SGD: S000005654) is phosphoribosylaminoimidazole carboxylase and catalyzes the reaction from AIR to CAIR in the IMP biosynthetic pathway.

[0020] In the present invention, ADE1 (system name: YAR015W, SGD: S000000070) is N-succinyl-5-aminoimidazole-4-carboxamide ribotide synthetase and catalyzes the reaction from CAIR to SAICAR in the IMP biosynthetic pathway.

[0021] In the present invention, ADE13 (system name: YLR359W, SGD: S000004351) is an adenylate succinate lyase that catalyzes the reaction from SAICAR to AICAR in the IMP biosynthesis pathway.

[0022] In this invention, ADE16 (strain name: YLR028C, SGD: S000004018) possesses the enzymatic activity of both 5-aminoimidazole-4-carboxamidoliponucleotide transformylase and inosine monophosphate cyclohydrolase, and ADE17 is a paralog resulting from whole-genome duplication.

[0023] In this invention, ADE17 (strain name: YMR120C, SGD: S000004727) possesses the enzymatic activity of both 5-aminoimidazole-4-carboxamidoliponucleotide transformylase and inosine monophosphate cyclohydrolase, and ADE16 is a paralog resulting from whole-genome duplication.

[0024] In the present invention, ADE3 (strain name: YGR204W, SGD: S000003436) has three enzymatic activities: 10-formyl-THF synthase, 5,10-methenyl-THF cyclohydrolase, and 5,10-methylene-THF dehydrogenase.

[0025] In the manufacturing method of the present invention, when fermentation is carried out using adenine-requiring yeast, the amount of purines and xanthine produced during fermentation is reduced compared to when fermentation is carried out using ordinary yeast, resulting in a reduced xanthine concentration in the fermented liquid. In other words, the purine concentration and xanthine concentration of the produced beer-flavored fermented beverage are reduced. In the beer-flavored fermented beverage of the present invention, the reduction in purine concentration and xanthine concentration can be confirmed, for example, by measuring and comparing the purine concentration and xanthine concentration in both cases: when fermentation is carried out using adenine-requiring yeast and when fermentation is carried out using ordinary yeast. Note that "xanthine" is one of the four purine bases (adenine, xanthine, guanine, and hypoxanthine) that make up purines.

[0026] Furthermore, in the production method of the present invention, when fermentation is carried out using adenine-requiring yeast, the ratio of xanthine content to purine content in the fermented liquid after fermentation is reduced compared to when fermentation is carried out using ordinary yeast. That is, the ratio of xanthine concentration to purine concentration in the produced beer-flavored fermented beverage is reduced. The reduction in the ratio of xanthine concentration to purine concentration in the beer-flavored fermented beverage of the present invention can be confirmed, for example, by calculating and comparing the ratio of xanthine concentration to purine concentration in both cases: when fermentation is carried out using the adenine-requiring yeast of the present invention and when fermentation is carried out using ordinary yeast.

[0027] The beer-flavored fermented beverage of the present invention has reduced concentrations of purines and / or xanthine. The upper limits (less than or equal to) of the purine concentration of the beer-flavored fermented beverage of the present invention are, for example, 35,000 ppb, 34,000 ppb, 33,000 ppb, 32,000 ppb, 31,000 ppb, 30,000 ppb, 29,000 ppb, 28,000 ppb, 27,000 ppb, 26,000 ppb, 25,000 ppb, 24,000 ppb, 23,000 ppb, 22,000 ppb, 21,000 ppb, and 2 It can be 0,000 ppb, 19,000 ppb, 18,000 ppb, 17,000 ppb, 16,000 ppb, 15,000 ppb, 14,000 ppb, 13,000 ppb, 12,000 ppb, 11,000 ppb, 10,000 ppb, 9,000 ppb, 8,000 ppb, 7,000 ppb, 6,000 ppb, 5,000 ppb, 4,000 ppb, 3,000 ppb, or 2,000 ppb. The lower limit (greater than or greater than) can be, for example, 0 ppb, 10 ppb, 50 ppb, 100 ppb, 200 ppb, 300 ppb, 400 ppb, 500 ppb, 600 ppb, 700 ppb, 800 ppb, 900 ppb, or 1,000 ppb. These upper and lower limits can be combined in any way. Beverages with a purine concentration of less than 5,000 ppb are considered "purine-free beverages." In this invention, the unit "ppb" is synonymous with "μg / L."

[0028] The upper limits (less than or equal to) of the xanthine concentration in the beer-flavored fermented beverage of the present invention are, for example, 20,000 ppb, 19,000 ppb, 18,000 ppb, 17,000 ppb, 16,000 ppb, 15,000 ppb, 14,000 ppb, 13,000 ppb, 12,000 ppb, 11,000 ppb, 10,000 ppb, 9,900 ppb, 9,800 ppb, 9,700 ppb, 9,600 ppb, 9,500 ppb, 9,400 ppb, 9,300 ppb, 9,200 ppb, 9,100 ppb, 9,000 ppb, and 8,000 ppb. ,900ppb, 8,800ppb, 8,700ppb, 8,600ppb, 8,500ppb, 8,400ppb, 8,300ppb, 8,200ppb, 8,100ppb, 8,000ppb, 7,900ppb, 7,800ppb, 7,700ppb, 7,600 ppb, 7,500ppb, 7,400ppb, 7,300ppb, 7,200ppb, 7,100ppb, 7,000ppb, 6,900ppb, 6,800ppb, 6,700ppb, 6,600ppb, 6,500ppb, 6,400ppb, 6,300ppb, 6 ,200ppb, 6,100ppb, 6,000ppb, 5,900ppb, 5,800ppb, 5,700ppb, 5,600ppb, 5,500ppb, 5,400ppb, 5,300ppb, 5,200ppb, 5,100ppb, 5,000ppb, 4,900 ppb, 4,800ppb, 4,700ppb, 4,600ppb, 4,500ppb, 4,400ppb, 4,300ppb, 4,200ppb, 4,100ppb, 4,000ppb, 3,900ppb, 3,800ppb, 3,700ppb, 3,600ppb, 3 It can be 500ppb, 3,400ppb, 3,300ppb, 3,200ppb, 3,100ppb, 3,000ppb, 2,900ppb, 2,800ppb, 2,700ppb, 2,600ppb, 2,500ppb, 2,400ppb, 2,300ppb, 2,200ppb, 2,100ppb, 2,000ppb, 1,900ppb, 1,800ppb, 1,700ppb, 1,600ppb, 1,500ppb, 1,400ppb, 1,300ppb, 1,200ppb, 1,100ppb, or 1,000ppb.The lower limit (greater than or greater than) can be, for example, 0 ppb, 10 ppb, 50 ppb, 100 ppb, 200 ppb, 300 ppb, 400 ppb, or 500 ppb. These upper and lower limits can be combined in any way.

[0029] In the beer-flavored fermented beverage of the present invention, the upper limit (less than or equal to) of the ratio of xanthine concentration (ppb) to purine concentration (ppb) can be, for example, 0.40, 0.39, 0.38, 0.37, 0.36, 0.35, 0.34, 0.33, 0.32, 0.31, 0.30, 0.29, 0.28, 0.27, 0.26, 0.25, 0.24, 0.23, 0.22, 0.21, 0.20, 0.19, 0.18, 0.17, 0.16, 0.15, 0.14, 0.13, 0.12, 0.11, or 0.10.

[0030] The concentrations of adenine, guanine, xanthine, and hypoxanthine can be measured by known methods. For example, they can be measured by detecting them using LC-MS / MS (liquid chromatography-mass spectrometry) after hydrolysis with perchloric acid (see Journal of Food Hygiene 55(2):110-116 (2014)). Furthermore, the purine concentration can be calculated as the total concentration of the four purine bases: adenine, xanthine, guanine, and hypoxanthine.

[0031] In the present invention, the purine concentration and xanthine concentration can be further reduced by known methods in addition to those described above. Non-limiting examples of methods for reducing the purine concentration and xanthine concentration include methods for reducing the purine content in beverages by contacting the pre-fermentation liquid or fermentation liquid with an adsorbent such as activated carbon or zeolite (see Japanese Patent Publication No. 2003-169658, Japanese Patent Publication No. 2004-290071, Japanese Patent Publication No. 2004-290072, Japanese Patent Publication No. 2015-112090, etc.), and raw materials other than malt that have low purine content (for example, soy protein). Methods for reducing purine content in beverages using corn grits (see Japanese Patent Publication No. 2014-117204, Japanese Patent Publication No. 2014-117205, etc.) and methods for reducing yeast-non-assimilable purines in wort by performing enzyme inactivation treatment to inactivate at least a portion of the enzyme group derived from malt, and then performing purine nucleosidase treatment on the brewing liquid (for example, Japanese Patent Publication No. 2018-64502). These methods for reducing purine concentration can be used individually or in combination.

[0032] In the present invention, the preparation of the mash can be carried out according to conventional methods, for example, by sequentially performing the following steps: (a) saccharifying a mixture of raw materials including malt and water, filtering it to obtain wort; (b) adding hops to the obtained wort and then boiling it; and (c) cooling the boiled wort. In the saccharification step, malt with high endoprotease activity and / or enzyme preparations possessing endoprotease activity can be used to promote protein degradation, and malt with low endoprotease activity can be used to suppress protein degradation.

[0033] In the present invention, the decomposition process can also be carried out by adding proteolytic enzymes, carbohydrate-degrading enzymes, and cellulose-degrading enzymes at any point before the completion of the fermentation process (for example, during mashing, during fermentation, or both during mashing and fermentation), either individually or in combination. Examples of proteolytic enzymes include protease preparations, examples of carbohydrate-degrading enzymes include enzymes derived from brewing raw materials such as malt, and commercially available enzyme preparations (such as α-amylase, β-amylase, glucoamylase, α-glucosidase, prunalase, and isoamylase), and examples of cellulose-degrading enzymes include commercially available enzyme preparations (such as β-glucanase, xylanase, and hemicellulase).

[0034] In the present invention, the saccharification temperature can be, for example, 30°C to 100°C, 30°C to 80°C, 30°C to 70°C, 35°C to 100°C, 35°C to 80°C, 35°C to 70°C, 40°C to 100°C, 40°C to 80°C, or 40°C to 70°C. The saccharification time can be set appropriately considering the saccharification temperature, but can be, for example, 30 minutes to 540 minutes, 30 minutes to 480 minutes, 30 minutes to 420 minutes, 30 minutes to 360 minutes, 30 minutes to 300 minutes, 30 minutes to 240 minutes, 30 minutes to 180 minutes, 50 minutes to 150 minutes, or 60 minutes to 140 minutes.

[0035] In the present invention, the boiling temperature can be, for example, 80°C to 150°C, 100°C to 150°C, 100°C to 140°C, or 100°C to 130°C. The boiling time can be set appropriately considering the boiling temperature, but can be, for example, 10 minutes to 240 minutes, 10 minutes to 180 minutes, 20 minutes to 150 minutes, or 30 minutes to 120 minutes.

[0036] In the present invention, the fermentation conditions are not particularly limited, except for the use of adenine-requiring yeast, and can be carried out according to conventional methods. The fermentation temperature can be, for example, 4°C to 30°C, 4°C to 25°C, 4°C to 20°C, 4°C to 18°C, 4°C to 15°C, 4°C to 12°C, 4°C to 10°C, 10°C to 30°C, 10°C to 25°C, 10°C to 20°C, or 10°C to 18°C. The fermentation time can be, for example, 1 to 20 days, 1 to 15 days, 1 to 10 days, 1 to 7 days, 4 to 20 days, 4 to 15 days, 4 to 10 days, or 4 to 7 days.

[0037] In the production of the beer-flavored fermented beverage of the present invention, brewing ingredients other than malt may be used. Usable brewing ingredients include unsprouted grains (e.g., unsprouted barley (including extract), unsprouted wheat (including extract)); rice, corn, sorghum, potatoes, starch, sugars (e.g., liquid sugar), fruits (e.g., dried fruit, boiled fruit, fruit juice, concentrated fruit juice), coriander or its seeds, spices or their raw materials (e.g., pepper, cinnamon, cloves, sansho pepper), herbs (e.g., chamomile, sage, basil, lemon). Ingredients include: auxiliary ingredients such as buckwheat, vegetables (for example, dried or boiled vegetables such as sweet potatoes and pumpkins), buckwheat or sesame, sugary substances (for example, honey, brown sugar), salt, miso, flowers, tea, coffee, cocoa (including preparations of tea, coffee, and cocoa), and seafood (for example, oysters, kelp, wakame seaweed, and bonito flakes); nitrogen sources such as protein hydrolysates and yeast extract; and other additives such as flavorings, colorings, foaming and foam retention enhancers, water conditioners, and fermentation aids.

[0038] In the production of the beer-flavored fermented beverage of the present invention, the raw materials used, other than brewing water, can be at least malt and hops, and in some cases sugars, rice, corn, starch, etc., can also be used as raw materials. It goes without saying that all-malt beer, among the beer-flavored fermented beverages produced, can be made from malt, hops, and water. The production method of the present invention can also be made without malt and / or ungerminated grains as raw materials, in which case the beer-flavored fermented beverage produced will have a malt usage ratio of 0%.

[0039] In the production of the beer-flavored fermented beverage of the present invention, malt and / or ungerminated grains can be used as at least part of the raw materials, preferably at least barley malt and wheat malt or both can be used as grain-derived raw materials, and malt extract can also be used. In the production of the beer-flavored fermented beverage of the present invention, the proportion of barley malt (mass) in the total mass of raw malt can be, for example, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 100% or less, or 100%, and the proportion of wheat malt (mass) in the total mass of raw malt can be, for example, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 100% or less, or 100%.

[0040] In the production of the beer-flavored fermented beverage of the present invention, malt and / or ungerminated grains can be used as at least part of the raw materials, preferably at least malt (e.g., wheat malt, barley malt) can be used as a grain-derived raw material, and malt extract can also be used. When malt is used as at least part of the raw materials in the production of the beer-flavored fermented beverage of the present invention, the malt usage ratio of the beverage can be, for example, 0%, 0% or more, 1% or less, 1% or more, 2% or less, 2% or more, 3% or less, 3% or more, 4% or more, 5% or less, 5% or more, 10% or less, 10% or more, 15% or less, 15% or more, 20% or less, 20% or more, 21% or less, 21% or more, 22% or less, 22% or more, 23% or less, 23% or more, 24% or less, 2 4% or more, 25% or less, 25% or more, 26% or less, 26% or more, 27% or less, 27% or more, 28% or less, 28% or more, 29% or less, 29% or more, 30% or less, 30% or more, 31% or less, 31% or more, 32% or less, 32% or more Above, 33% or less, 33% or more, 34% or less, 34% or more, 35% or less, 35% or more, 36% or less, 36% or more, 37% or less, 37% or more, 38% or less, 38% or more, 39% or less, 39% or more, 40% or less, 40% or more, 41 % or less, 41% or more, 42% or less, 42% or more, 43% or less, 43% or more, 44% or less, 44% or more, 45% or less, 45% or more, 46% or less, 46% or more, 47% or less, 47% or more, 48% or less, 48% or more, 49% or less , 49% or more, 50% or less, 50% or more, 55% or less, 55% or more, 60% or less, 60% or more, 65% or less, 65% or more, 70% or less, 70% or more, 75% or less, 75% or more, 80% or less, 80% or more, 81% or less, 81% The percentages can be 82% or less, 82% or more, 83% or less, 83% or more, 84% or less, 84% or more, 85% or less, 85% or more, 90% or less, 90% or more, 95% or less, 95% or more, 100% or less, or 100%, preferably 50% or more, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, or 85% or more, and "or less" can be written as "less than", and "or more" can be written as "greater than".These upper and lower limits can be combined in any way, for example: 1% to 100%, 1% to 90%, 1% to 85%, 1% to 84%, 1% to 83%, 1% to 82%, 1% to 81%, 1% to 80%, 1% to 70%, 1% to 60%, 1% to 50%, 10% to 100%, 10% to 90%, 10% to 85%, 10% to 84%, 10% or more. 83% or less, 10% to 82%, 10% to 81%, 10% to 80%, 10% to 70%, 10% to 60%, 10% to 50%, 20% to 100%, 20% to 90%, 20 % to 85%, 20% to 84%, 20% to 83%, 20% to 82%, 20% to 81%, 20% to 80%, 20% to 70%, 20% to 60%, 20% to 50%, 30% to 100%, 30% to 90%, 30% to 85%, 30% to 84%, 30% to 83%, 30% to 82%, 30% to 81%, 30% to 80%, 30% to 70% Below, 30% to 60%, 30% to 50%, 40% to 100%, 40% to 90%, 40% to 85%, 40% to 84%, 40% to 83%, 40% to 82%, 40% or more The malt content can be 81% or less, 40% to 80%, 40% to 70%, 40% to 60%, 40% to 50%, 50% to 100%, 50% to 90%, 50% to 85%, 50% to 84%, 50% to 83%, 50% to 82%, 50% to 81%, 50% to 80%, 50% to 75%, 50% to 70%, 50% to 65%, or 50% to 60%. In this invention, "malt usage ratio" refers to the ratio of the mass of malt to the mass of all raw materials excluding hops and brewing water.

[0041] The beer-flavored fermented alcoholic beverage of the present invention allows for the arbitrary setting of the alcohol concentration (ethanol concentration), but the lower limit of the alcohol concentration (greater than or equal to) is, for example, 1.0v / v%, 1.1v / v%, 1.2v / v%, 1.3v / v%, 1.4v / v%, 1.5v / v%, 1.6v / v%, 1.7v / v%, 1.8v / v%, 1.9v / v%, 2.0v / v%, 2.1v / v%, 2.2v / v%, 2.3v / v%, 2.4v / v%, 2.5v / v%, 2.6v / v%, 2.7v / v%, 2.8v / v%, 2.9 The values ​​can be v / v%, 3.0v / v%, 3.1v / v%, 3.2v / v%, 3.3v / v%, 3.4v / v%, 3.5v / v%, 3.6v / v%, 3.7v / v%, 3.8v / v%, 3.9v / v%, 4.0v / v%, 4.1v / v%, 4.2v / v%, 4.3v / v%, 4.4v / v%, or 4.5v / v%, with upper limits (less than or equal to) being, for example, 20.0v / v%, 19.5v / v%, 19.0v / v%, 18.5v / v%, 18.0v / v%, 17.5v / v%, 17.0v / v%, 16.5v / v%, 16.0v / v%, 15.5v / v%, 15.0v / v%, 14.5v / v%, 14.0v / v%, 13.5v / v%, 13.0v / v%, 12.5v / v%, 12.0v / v%, 11.5v / v%, 11.0v / v%, 10.5v / v%, 10.0v / v%, 9 .9v / v%, 9.8v / v%, 9.7v / v%, 9.6v / v%, 9.5v / v%, 9.4v / v%, 9.3v / v%, 9.2v / v%, 9.1v / v%, 9.0v / v%, 8.9v / v%, 8.8v / v%, 8.7v / v%, 8.6v / v%, 8.5v / v% , 8.4v / v%, 8.3v / v%, 8.2v / v%, 8.1v / v%, 8.0v / v%, 7.9v / v%, 7.8v / v%, 7. 7v / v%, 7.6v / v%, 7.5v / v%, 7.4v / v%, 7.3v / v%, 7.2v / v%, 7.1v / v%, 7.0v / v%, 6.9v / v%, 6.8v / v%, 6.7v / v%, 6.6v / v%, 6.5v / v%, 6.4v / v%, 6.3v / v%, 6.2v / v%, 6.1v / v%, 6.0v / v%, 5.9v / v%, 5.8v / v%, 5.7v / v%, 5.6v / v% or 5.The alcohol concentration can be set to 5 v / v%. These lower and upper limits can be combined arbitrarily, and the alcohol concentration of the beer-flavored fermented alcoholic beverage of the present invention can be, for example, 1.0 v / v% to 20.0 v / v%, 1.0 v / v% to 19.0 v / v%, 1.0 v / v% to 18.0 v / v%, 1.0 v / v% to 17.0 v / v%, 1.0 v / v% to 16.0 v / v%, 1.0 v / v% to 15.0 v / v%, 1.0 v / v% to 14.0 v / v%, 1.0 v / v% to 13.0 v / v%, or 1.0 v / v% to 12.0 v / v%. Below, 1.0v / v% to 11.0v / v%, 1.0v / v% to 10.0v / v%, 1.0v / v% to 9.0v / v%, 1.0v / v% to 8.0v / v%, 1.0v / v% to 7.0v / v%, 1.0v / v% to 6.0v / v%, 2.0 v / v% to 20.0v / v%, 2.0v / v% to 19.0v / v%, 2.0v / v% to 18.0v / v%, 2.0v / v% to 17.0v / v%, 2.0v / v% to 16.0v / v%, 2.0v / v% to 15.0v / v%, 2.0v / v% to 14.0v / v%, 2.0v / v% to 13.0v / v%, 2.0v / v% to 12.0v / v%, 2.0v / v% to 11.0v / v%, 2.0v / v% to 10.0v / v%, 2.0v / v% to 9.0v / v%, 2.0v / v% and above Upper 8.0v / v% or less, 2.0v / v% or more and 7.0v / v% or less, 2.0v / v% or more and 6.0v / v% or less, 3.0v / v% or more and 20.0v / v% or less, 3.0v / v% or more and 19.0v / v% or less, 3.0v / v% or more and 18.0v / v% or less, 3.0v / v% or more and 17.0 v / v% or less, 3.0v / v% or more and 16.0v / v% or less, 3.0v / v% or more and 15.0v / v% or less, 3.0v / v% or more and 14.0v / v% or less, 3.0v / v% or more and 13.0v / v% or less, 3.0v / v% or more and 12.0v / v% or less, 3.0v / v% or more and 11.0v / v% or less, 3.0v / v% or more and 10.0v / v% or less, 3.0v / v% or more and 9.0v / v% or less, 3.0v / v% or more and 8.0v / v% or less, 3.0v / v% or more and 7.0v / v% or less, 3.0v / v% or more and 6.0v / v% or less, 4.0v / v% or more and 20.0v / v% or less, 4.0v / v% or more and 19.0v / v% or less, 4.0v / v% or more and 18.0v / v% or less, 4.0v / v% or more and 17.0v / v% or less, 4.0v / v% or more and 16.0 v / v% or less, 4.0v / v% or more and 15.0v / v% or less, 4.0v / v% or more and 14.0v / v% or less, 4.0v / v% or more and 13.0v / v% or less, 4.0v The values ​​can be between 12.0v / v% and 12.0v / v%, between 4.0v / v% and 11.0v / v%, between 4.0v / v% and 10.0v / v%, between 9.0v / v%, between 8.0v / v%, between 7.0v / v%, or between 6.0v / v%.

[0042] The alcohol concentration (ethanol concentration) of beer-flavored fermented alcoholic beverages can be measured by methods widely known to those skilled in the art, for example, by the method described in the "National Tax Agency's prescribed analytical method."

[0043] In the beer-flavored fermented non-alcoholic beverage of the present invention, the alcohol (ethanol) concentration can be less than 1.00 v / v%, and can be 0.99 v / v% or less, 0.95 v / v% or less, 0.90 v / v% or less, 0.85 v / v% or less, 0.80 v / v% or less, 0.75 v / v% or less, 0.70 v / v% or less, 0.65 v / v% or less, 0.60 v / v% or less, 0.55 v / v% or less, 0.50 v / v% or less, 0.45 v / v% or less, 0.40 v / v% or less, 0.35 v / v% or less, 0.30 v / v% or less, 0.25 v / v% or less, 0. The limits can also be 20 v / v% or less, 0.15 v / v% or less, 0.10 v / v% or less, 0.05 v / v% or less, 0.01 v / v% or less, and further, concentrations less than 0.01 v / v% (0.00 v / v%) can be 0.009 v / v% or less, 0.005 v / v% or less, 0.004 v / v% or less, 0.003 v / v% or less, 0.002 v / v% or less, or 0.001 v / v% or less, and the lower limits (greater than or equal to) can be 0.001 v / v%, 0.002 v / v%, 0.003 v / v%, or 0.004 v / v%. These upper and lower limits can be combined arbitrarily, "less than or equal to" can be changed to "less than", and "greater than or equal to" can be changed to "greater than".

[0044] The alcohol concentration (ethanol concentration) of beer-flavored fermented non-alcoholic beverages can be measured by gas chromatography (GC) with a flame ionization detector (FID). For more accurate concentration measurement, it is desirable to use a calibration curve created based on the measured values ​​of several control samples with known concentrations. These control samples with known concentrations are preferably within the same concentration range as the one being measured. Furthermore, it is preferable to use an internal standard; an example of an internal standard is 2-propanol.

[0045] In the production of a beer-flavored fermented non-alcoholic beverage of the present invention, methods for removing alcohol from a beer-flavored fermented alcoholic beverage include, for example, (i) a method of removing alcohol and low-boiling point components by distillation under reduced pressure or atmospheric pressure, (ii) a method of removing alcohol and low-molecular-weight components using a reverse osmosis (RO) membrane, and (iii) a method of removing volatile components by adsorption to vapor using centrifugal force. A beer-flavored fermented non-alcoholic beverage obtained by such a method is called a de-alcoholized beverage, and can also be described as a beverage made from de-alcoholized wort ferment.

[0046] The pH (measured at 25°C) of the beer-flavored fermented beverage of the present invention is not limited, but its lower limit (greater than or equal to) can be 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, or 3.0, and its upper limit (less than or equal to) can be 5.0, 4.9, 4.8, 4.7, 4.6, 4.5, 4.4, or 4.3. These lower and upper limits can be combined in any way, for example, 2.0 to 5.0, 2.0 to 4.5, 2.5 to 5.0, 2.5 to 4.5, 3.0 to 5.0, or 3.0 to 4.5. The pH of the beer-flavored fermented beverage of the present invention can be adjusted using a pH adjusting agent. The pH of the beer-flavored fermented beverage can be measured using a commercially available pH meter (e.g., a benchtop pH meter, manufactured by Horiba, Ltd.).

[0047] The beer-flavored fermented beverage of the present invention can be served as a carbonated beverage. The carbon dioxide pressure (gas pressure at 20°C) of the beer-flavored fermented beverage of the present invention is not limited, but its lower limit (greater than or equal to) can be 0.05 MPa, 0.06 MPa, 0.07 MPa, 0.08 MPa, 0.09 MPa, or 0.1 MPa, and its upper limit (less than or equal to) can be 0.4 MPa, 0.39 MPa, 0.38 MPa, 0.37 MPa, 0.36 MPa, or 0.35 MPa. These upper and lower limits can be combined in any way, for example, 0.05 MPa or more and 0.4 MPa or less, 0.07 MPa or more and 0.38 MPa or less, or 0.1 MPa or more and 0.35 MPa or less.

[0048] The bitterness value (BU) of the beer-flavored fermented beverage of the present invention is not limited, but its lower limit (greater than or equal to) can be, for example, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15, and its upper limit (less than or equal to) can be, for example, 120, 115, 110, 100, 95, 90, 85, 80, 75, 70, 65, 60, 59, 58, 57, 56, 55, 54, 53, 52, 51, 50, 49, 48, 47, 46, 45, 44, 43, 42, 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, or 20.These lower and upper limits can be combined in any way, for example, 5 to 120, 5 to 110, 5 to 100, 5 to 90, 5 to 80, 5 to 70, 5 to 60, 5 to 55, 5 to 50, 5 to 45, 5 to 40, 5 to 35, 5 to 30, 5 to 25, 5 to 20, 6 to 120, 6 to 110, 6 to 100, 6 to 90, 6 to 80, 6 to 70, 6 to 60, 6 to 55, 6 to 50, 6 to 45, 6 to 40, 6 to 35, 6 to 30, 6 to 25, 6 to 20, 7 to 120, 7 to 110, 7 7 to 100, 7 to 90, 7 to 80, 7 to 70, 7 to 60, 7 to 55, 7 to 50, 7 to 45, 7 to 40, 7 to 35, 7 to 30, 7 to 25, 7 to 2 0 or less, 8 to 120, 8 to 110, 8 to 100, 8 to 90, 8 to 80, 8 to 70, 8 to 60, 8 to 55, 8 to 50, 8 to 45, 8 to 40, 8 to 3 5 or less, 8 to 30, 8 to 25, 8 to 20, 9 to 120, 9 to 110, 9 to 100, 9 to 90, 9 to 80, 9 to 70, 9 to 60, 9 to 55, 9 to 50 The bitterness value of the beer-flavored fermented beverage of the present invention can be adjusted by the hop variety, amount added, timing of addition, etc. The bitterness value of the beer-flavored beverage can be measured by the method described in the "National Tax Agency's prescribed analytical method".

[0049] The appearance extract (°P) of the beer-flavored fermented beverage of the present invention is not limited, but its lower limit (greater than or equal to) can be, for example, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, or 7.5, and its upper limit (less than or equal to) can be, for example, 15.0, 14.5, 14.0, 13.5, 13.0, 12.5, 12.0, 11.5, 11.0, 10.5, or 10.0. These lower and upper limits can be combined in any way, for example, 2 to 15, 2 to 14, 2 to 13, 2 to 12, 2 to 11, 2 to 10, 3 to 15, 3 to 14, 3 to 13, 3 to 12, 3 to 11, 3 to 10, 4 to 15, 4 to 14, 4 to 13, 4 to 12, 4 to 11, 4 to 10, 5 to 15, 5 to 14, 5 to 14, 5 to 13, 5 to 12, 5 to 11, 5 to 10, 6 to 15, 6 to 14, 6 to 13, 6 to 12, 6 to 11, 6 to 10, 7 to 15, 7 to 14, 7 to 13, 7 to 12, 7 to 11, or 7 to 10. The visual extract of the beer-flavored fermented beverage of the present invention can be adjusted by the amount of raw materials used and the fermentation and filtration conditions. The visual extract of the beer-flavored beverage can be calculated using the BCOJ Beer Analysis Method (Brewing Brewers Association, 8.3.6 (Alcoholizer Method)) using the specific gravity and alcohol content measured by the BCOJ Beer Analysis Method (Brewing Brewers Association, 8.5 (Extract-Related Calculation Method)).

[0050] The wort extract (°P) of the beer-flavored fermented beverage of the present invention is not limited, but its lower limit (greater than or equal to) can be, for example, 5.0, 5.5, 6.0, 6.5, 7.0, or 7.5, and its upper limit (less than or equal to) can be, for example, 50.0, 45.0, 40.0, 35.0, 30.0, 25.0, 20.0, 15.0, or 14.0. These lower and upper limits can be combined in any way, for example, 5.0 to 50.0, 5.0 to 45.0, 5.0 to 40.0, 5.0 to 35.0, 5.0 to 30.0, 5.0 to 25.0, 5.0 to 20.0, 5 to 15.0, 5 to 14.0, 6.0 to 50.0, 6.0 to 45.0, 6.0 to 40.0, 6.0 to 35.0, 6.0 to 30.0, 6.0 to 25.0, 6.0 to 20.0, 6.0 to 15.0, or 6.0 to 14.0. The wort extract of the beer-flavored fermented beverage of the present invention can be adjusted by the amount of raw materials used and the fermentation and filtration conditions. The wort extract of beer-flavored beverages can be measured according to the BCOJ Beer Analysis Method (Japan Beer Brewers Association, 8.5 (Calculation Method for Extracts)).

[0051] The beer-flavored fermented beverage of the present invention can be provided as a packaged beverage. The container used for the beverage according to the present invention may be any container that is normally used for filling beverages, such as metal cans, barrels, plastic bottles (e.g., PET bottles, cups), paper containers, bottles, pouches, etc., but preferably metal cans / barrels, plastic bottles (e.g., PET bottles), or bottles.

[0052] In addition to carbon dioxide, nitrogen may also be added to the beer-flavored fermented beverage of the present invention. The nitrogen concentration can be adjusted as appropriate according to preference. The form of nitrogen used when adding nitrogen to the beverage may be any form known to those skilled in the art, such as nitrogen gas or liquid nitrogen, but liquid nitrogen is preferred. The total gas pressure including nitrogen in the beer-flavored fermented beverage of the present invention can be adjusted within the range of 0.05 to 0.4 MPa (gas pressure at 20°C) (without separately sealed nitrogen). There are many purposes for adding nitrogen, but for example, it can make the foam of the beer-flavored fermented beverage creamier.

[0053] Furthermore, the container used for the beer-flavored fermented beverage of the present invention may include a hollow insert, such as a so-called "widget," which is separate from the container and used to efficiently supply nitrogen or the like to the beverage inside the container. The shape of the widget can vary, such as a sphere or a cube, and any shape is acceptable. The widget may or may not be fixed to the container. Before the container is opened, the widget contains a gas containing pressurized nitrogen gas that is in equilibrium with the total pressure of the container, which is higher than the atmospheric pressure. When the container is opened to make the beverage drinkable, the empty space where there is no beverage is released, resulting in a pressure difference, and the nitrogen sealed inside the widget is ejected into the container. It is desirable that the container and the widget be arranged such that when the container is opened, the nitrogen inside the widget is ejected directly into the liquid.

[0054] Furthermore, the method for incorporating nitrogen into the beer-flavored fermented beverage of the present invention can include methods such as using small objects to eject nitrogen, or, in addition to nitrogen already being present in the beverage, adding liquid nitrogen or the like to the container to fill the empty space with nitrogen and saturate the beverage with nitrogen. This addition of liquid nitrogen can be done simultaneously when sealing the beer-flavored beverage in the container. According to these methods, the so-called cascade foam phenomenon occurs in the beer-flavored fermented beverage of the present invention, making it possible to generate more distinctive foam.

[0055] According to another aspect of the present invention, a method for reducing the xanthine concentration and / or purine concentration of a beer-flavored fermented beverage is provided, comprising the step of fermentation using adenine-requiring yeast in the production of the beer-flavored fermented beverage. The method of the present invention can be carried out according to the description of the beer-flavored fermented beverage and the method for producing the same of the present invention. [Examples]

[0056] The present invention will be described more specifically based on the following examples, but the present invention is not limited to these examples.

[0057] Measurement of alcohol concentration The alcohol concentration (ethanol concentration) was measured using the method described in the "National Tax Agency's prescribed analytical method."

[0058] Measurement of purine concentration Adenine, guanine, xanthine, and hypoxanthine concentrations were measured using LC-MS / MS (liquid chromatography-mass spectrometry) after hydrolysis with perchloric acid (see Journal of Food Hygiene 55(2):110-116(2014)). Purine concentration was calculated as the total concentration of the four purine bases: adenine, xanthine, guanine, and hypoxanthine.

[0059] Calculation of the external extract The visible extract was calculated using the BCOJ Beer Analysis Method (Bergary Brewers Association, 8.3.6 (Alcoholizer Method)), with the specific gravity and alcohol content measured according to the BCOJ Beer Analysis Method (Bergary Brewers Association, 8.5 (Extract-Related Calculation Method)).

[0060] Example 1: Investigation of xanthine reduction using adenine-requiring yeast (1) In Example 1, we investigated the effect on xanthine concentration after fermentation by adenine-requiring yeast.

[0061] (1) Method a. Preparation of wort Wort (malt usage ratio 41.6%) was prepared using a raw material containing spray malt (Spraymalt Muntons extra light 8 EBC) and glucose. Wort (malt usage ratio 81.6%) was also prepared using a raw material containing spray malt and glucose. Furthermore, wort (malt usage ratio 100%) was prepared using spray malt as the raw material. Each wort was obtained by mixing the raw materials with water, dissolving them at 98°C, and then filtering; the visible extract was 11°P.

[0062] (i) Yeast and fermentation conditions Add yeast to each 500 mL of wort obtained in step A above. Saccharomyces pastorianus After adding () and carrying out fermentation, the xanthine concentration of the fermentation liquid was measured and the purine concentration was calculated. Four types of yeast were used: lager yeast (1) and (2), adenine-requiring yeast (1) (lamger yeast (1) mutant), and adenine-requiring yeast (2) (lamger yeast (2) mutant). The fermentation temperature was 15°C and the fermentation period was 7 days.

[0063] Genetic analysis of adenine-requiring yeast DNA was extracted using the DNeasy Blood & Tissue Kit (QIAGEN), whole-genome sequencing was performed using NovaSeq6000 (Illumina), and the genes of adenine-requiring yeast were analyzed using Geneious Prime (Dotmatics).

[0064] (2) Results The results are shown in Tables 1-3. At malt usage ratios of 41.6% and 81.6%, the xanthine concentration in the fermented liquid after fermentation was approximately half or less than that when using conventional yeast when adenine-requiring yeast was used. On the other hand, at a malt usage ratio of 100%, the xanthine concentration in the fermented liquid after fermentation was lower when using adenine-requiring yeast than when using conventional yeast, but the difference was not significant. Note that in Tables 1-3, the purine and xanthine concentrations represent the average values ​​for each test group.

[0065] Furthermore, genetic analysis of adenine-requiring yeast (1) and (2) revealed mutations in the sequences encoding the AIRS domain of the ADE5,7 genes, and confirmed that the 604th amino acid of the produced ADE5,7 proteins was substituted from asparagine to lysine.

[0066] The results above indicate that when adenine-requiring yeast is used, the xanthine concentration in the fermented liquid after fermentation is reduced compared to when conventional yeast is used.

[0067] [Table 1]

[0068] [Table 2]

[0069] [Table 3]

[0070] Example 2: Investigation of xanthine reduction using adenine-requiring yeast (2) In Example 2, the amounts of metabolites in the IMP biosynthesis pathway of adenine-requiring yeast were measured.

[0071] (1) Method Metabolites of the IMP biosynthesis pathway were quantified using an Agilent / HMT metabolome analysis system (Agilent, G7100A CE Instrument, G6230B TOF LC / MS) after 40 hours of stirring culture of adenine-requiring yeast (1) or (2) from Example 1 in SD medium (Becton Dickinson, Inc.) supplemented with 1 ppm adenine hydrochloride. The quantification conditions were set according to the description in Soga T, et al., J Proteome Res. 2003 Sep-Oct;2(5):488-94. In addition, 100 μM of D-Camphar-10-sulfonic acid sodium salt (Fujifilm Wako Pure Chemical Industries, Ltd., 037-01032) was added to the sample as an internal standard.

[0072] The amount of each metabolite was calculated by dividing the peak area obtained from the chromatogram of each metabolite, obtained using the Agilent / HMT metabolome analysis system described above, by the peak area of ​​the internal standard substance (internal standard ratio).

[0073] (2) Results The results are shown in Table 4. Compared to normal yeast, adenine-requiring yeast showed accumulation of FGAM, while AIR was not detected. This result indicates that in adenine-requiring yeast, there is a mutation in the sequence encoding the AIRS domain of the ADE5,7 genes, resulting in non-functional enzymatic activity of the AIRS domain of the ADE5,7 protein. Although not bound by the following theory, it is thought that the non-functional enzymatic activity of the AIRS domain of the ADE5,7 protein in adenine-requiring yeast prevented the IMP biosynthesis pathway from functioning, resulting in a decrease in the xanthine concentration of the fermented broth after fermentation. Note that in Table 4, the amounts of FGAM and AIR represent the average values ​​for each test group.

[0074] [Table 4]

[0075] Example 3: Investigation of xanthine reduction using adenine-requiring yeast (3) In Example 3, we investigated the effect of ADE5,7 gene knockout on xanthine concentration after fermentation in yeast.

[0076] (1) Method a. Preparation of wort The raw materials, including spray malt (Spraymalt Muntons extra light 8 EBC) and glucose, were mixed with water and dissolved at 98°C. The mixture was then filtered to obtain wort (malt usage ratio 41.6%, visible extract 11°P).

[0077] (i) Yeast and fermentation conditions Add yeast to 30 mL of each wort obtained in A above ( Saccharomyces cerevisiae After adding the specified ingredients and allowing fermentation to proceed, the purine and xanthine concentrations of the fermentation liquid were measured. The yeasts used were common yeast (BY4741) and adenine-requiring yeast S. cerevisiae Knock Out (YGL234W) (purchased from Horizon Discovery (Japanese distributor Funakoshi Co., Ltd.)). The fermentation temperature was 15°C or 20°C, and the fermentation period was 7 days.

[0078] (2) Results The results are shown in Tables 5 and 6. At both fermentation temperatures of 15°C and 20°C, when yeast with the ADE5,7 genes knocked out was used as an adenine-requiring yeast, the xanthine concentration in the fermentate after fermentation was reduced compared to when normal yeast was used. These results support the idea that the xanthine concentration in the fermentate after fermentation is reduced in adenine-requiring yeast because the de novo synthesis pathway does not function.

[0079] [Table 5]

[0080] [Table 6]

Claims

1. A method for producing a beer-flavored fermented beverage, comprising the step of fermenting using adenine-requiring yeast, wherein the adenine-requiring yeast is deficient in at least a portion of the function of the enzyme that catalyzes the metabolism of the IMP biosynthesis pathway.

2. The manufacturing method according to claim 1, wherein the enzyme is one or more selected from the group consisting of ADE5, ADE7, ADE4, ADE8, ADE6, ADE2, ADE1, ADE13, ADE16, ADE17, and ADE3.

3. The manufacturing method according to claim 1, wherein the enzymes are ADE5 and ADE7.

4. The manufacturing method according to claim 1 or 2, wherein the malt usage ratio of the beverage is 50% or more and 90% or less.

5. The manufacturing method according to claim 1 or 2, wherein the beverage has a xanthine concentration of 18,000 ppb or less and / or a purine concentration of 35,000 ppb or less.

6. The manufacturing method according to claim 1 or 2, wherein the ratio of xanthine concentration (ppb) to purine concentration (ppb) of the beverage is 0.4 or less.

7. A beer-flavored fermented beverage having a xanthine concentration of 18,000 ppb or less and / or a purine concentration of 35,000 ppb or less, and containing adenine-requiring yeast.

8. The beverage according to claim 7, wherein the ratio of xanthine concentration (ppb) to purine concentration (ppb) is 0.4 or less.

9. The beverage according to claim 7 or 8, wherein the malt usage ratio is 50% or more and 90% or less.

10. A method for reducing the xanthine concentration and / or purine concentration of a beer-flavored fermented beverage, comprising the step of fermentation using adenine-requiring yeast, wherein the adenine-requiring yeast is deficient in at least part of the function of the enzyme that catalyzes the metabolism of the IMP biosynthesis pathway.

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