Method for manufacturing beer-flavored beverages

Abstract

To provide a method for producing a beer-flavored beverage with reduced purine content by breaking down nucleosides while minimizing the number of processes and energy costs. [Solution] A method for producing a beer-flavored beverage, comprising the step (A) of adding a microorganism having nucleoside assimilation ability to a wort fermentation liquid.

Description

Technical Field

[0001] The present invention relates to a method for producing 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 content has been increasing. Purine bodies are a general term for compounds having a purine skeleton. 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. It is known that nucleosides such as adenosine, guanosine, inosine, and xanthosine are hydrolyzed by the action of nucleosidase and become purine bases such as adenine, guanine, hypoxanthine, and xanthine, respectively, and the content is reduced. Among the purine bases, adenine, guanine, and hypoxanthine are known to be assimilated as nitrogen sources by common brewing yeasts (for example, yeasts belonging to the genus Saccharomyces) in the beer brewing process, and the content is reduced.

[0003] Patent Document 1 describes a method for producing a fermented beer-like foaming beverage by allowing nucleoside phosphorylase and / or nucleosidase to act on wort, then performing boiling treatment to inactivate the enzyme, and then further inoculating yeast for fermentation.

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0005] However, the method described in Patent Document 1 requires boiling (heat treatment) to inactivate nucleosidase, resulting in a large number of steps in the production of beer-flavored beverages and high energy costs associated with the heat treatment. Therefore, there was a need for a method that could decompose nucleosides and produce beer-flavored beverages with reduced purine content while reducing the number of steps and energy costs.

[0006] The present invention aims to provide a method for producing a beer-flavored beverage with reduced purine content by decomposing nucleosides while minimizing the number of steps and energy costs. [Means for solving the problem]

[0007] The inventors of the present invention have discovered that by adding microorganisms capable of assimilating nucleosides to the wort fermentation liquid in step (A), the microorganisms capable of assimilating nucleosides can be removed by filtration, thereby enabling the production of a beer-flavored beverage with reduced purine content by decomposing nucleosides while keeping the number of steps and energy costs down. This led to the invention of the method for producing a beer-flavored beverage.

[0008] In other words, the present invention relates to a method for producing the following beer-flavored beverage, although it is not limited to the following. [1] A method for producing a beer-flavored beverage, comprising the step (A) of adding a microorganism having nucleoside assimilation ability to a wort fermentation liquid. [2] The method for producing a product according to [1] above, wherein in step (A), a microorganism having nucleoside assimilation ability assimilates a nucleoside. [3] The method for producing the microorganism having nucleoside assimilation ability is a yeast having nucleoside assimilation ability, as described in [1] or [2] above. [4] The method for producing a nucleoside, wherein the yeast having nucleoside assimilation ability is at least one selected from the group consisting of yeasts belonging to the genus Lachancea, yeasts belonging to the genus Zygosaccharomyces, and yeasts belonging to the genus Kluyveromyces. [5] The method for producing the product according to [3] above, wherein the yeast having nucleoside assimilation ability is at least one selected from the group consisting of Lachancea thermotolerans, Lachancea fermentati, Zygosaccharomyces rouxii, and Kluyveromyces marxianus. [6] A method of manufacturing according to any of [1] to [5] above, excluding the step of adding nucleosidase (B). [Effects of the Invention]

[0009] According to the method for producing beer-flavored beverages of the present invention, it is possible to produce beer-flavored beverages with reduced purine content by decomposing nucleosides while reducing the number of steps and energy costs. [Modes for carrying out the invention]

[0010] The method for producing a beer-flavored beverage of the present invention (hereinafter also simply referred to as the "production method of the present invention") includes step (A) of adding microorganisms having nucleoside assimilation ability to a wort fermentation liquid. According to the production method of the present invention, by performing step (A), it is possible to produce a beer-flavored beverage with reduced purine content by decomposing nucleosides while reducing the number of steps and energy costs.

[0011] <Process (A)> Step (A) is the step of adding microorganisms capable of assimilating nucleosides to the wort fermentation liquid.

[0012] In this specification, "wort ferment liquor" refers to the liquid from which the components of malt have been dissolved and which has undergone fermentation by brewing yeast. "Wort ferment liquor" is a concept that includes fermented beer-flavored beverages. Also in this specification, "wort" refers to the liquid from which the components of malt have been dissolved and which has not undergone fermentation by brewing yeast. "Wort" is a concept that includes the saccharified liquid obtained in the saccharification process. In this specification, "wort ferment liquor" and "wort" are clearly distinguished.

[0013] In the production method of the present invention, the total nitrogen content of the wort ferment is not particularly limited, but is preferably 2 to 100 mg / 100 mL, more preferably 5 to 80 mg / 100 mL, and even more preferably 10 to 50 mg / 100 mL. In this specification, "total nitrogen content" refers to the total amount of all nitrogen compounds, including proteins and compounds containing amino acids. The total nitrogen content of the wort ferment 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). The total nitrogen content of the wort fermentation liquid 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 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, 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.), and the beer filtration conditions.

[0014] In this specification, nucleoside assimilation means hydrolyzing nucleosides (adenosine, guanosine, inosine, and xanthosine) among purines to produce purine bases (adenine, guanine, hypoxanthine, and xanthine). In this specification, a microorganism having nucleoside assimilation ability means a microorganism that has the ability to hydrolyze at least one selected from the group consisting of adenosine, guanosine, inosine, and xanthosine to produce at least one selected from the group consisting of adenine, guanine, hypoxanthine, and xanthine. The microorganisms possessing nucleoside assimilation ability are not particularly limited as long as they are microorganisms capable of assimilating nucleosides, and may include eukaryotes such as fungi like yeast, or prokaryotes such as bacteria and archaea, but yeast possessing nucleoside assimilation ability is preferred. One type of microorganism possessing nucleoside assimilation ability may be used alone, or two or more types may be used in combination.

[0015] Examples of yeasts that possess nucleoside assimilation ability include those belonging to the genera Lachancea, Zygosaccharomyces, Kluyveromyces, and Torulaspora. Examples of yeasts belonging to the genus Lachancea include Lachancea fermentati and Lachancea thermotolerans. Examples of yeasts belonging to the genus Zygosaccharomyces include Zygosaccharomyces rouxii. Examples of yeasts belonging to the genus Kluyveromyces include Kluyveromyces marxianus. Examples of yeasts belonging to the genus Torulaspora include Torulaspora delbrueckii and Torulaspora pretoriensis. In the production method of the present invention, the yeast having nucleoside assimilation ability is preferably at least one selected from the group consisting of yeasts belonging to the genus Lachancea, yeasts belonging to the genus Zygosaccharomyces, yeasts belonging to the genus Kluyveromyces, and yeasts belonging to the genus Torulaspora; more preferably at least one selected from the group consisting of yeasts belonging to the genus Lachancea, yeasts belonging to the genus Zygosaccharomyces, and yeasts belonging to the genus Kluyveromyces; and even more preferably yeasts belonging to the genus Lachancea or yeasts belonging to the genus Zygosaccharomyces. In the production method of the present invention, the yeast having nucleoside-assimilating ability is preferably at least one selected from the group consisting of Lachancea fermentati, Lachancea thermotolerans, Zygosaccharomyces rouxii, Kluyveromyces marxianus, and Torulaspora delbrueckii, more preferably at least one selected from the group consisting of Lachancea fermentati, Lachancea thermotolerans, Zygosaccharomyces rouxii, and Kluyveromyces marxianus, and still more preferably at least one selected from the group consisting of Lachancea fermentati, Lachancea thermotolerans, and Zygosaccharomyces rouxii.

[0016] As the microorganism having nucleoside-assimilating ability, a commercially available product may be used, or one isolated and purified from nature may be used. Further, these microorganisms having nucleoside-assimilating ability may be grown in advance by preculture, and the obtained preculture solution may be used in step (A). Furthermore, when the yeast is dried, it may be rehydrated by a conventionally known method and then used in step (A).

[0017] When pre-culturing a microorganism having nucleoside assimilation ability, the culture conditions are not particularly limited as long as the microorganism having nucleoside assimilation ability can grow, and can be appropriately set according to the type of the microorganism having nucleoside assimilation ability and the like. The culture temperature may be, for example, 10 to 35 °C, preferably 20 to 30 °C. Also, the culture time may be, for example, 5 hours to 5 days, preferably 16 hours to 3 days.

[0018] In step (A), it is preferable to allow a microorganism having nucleoside assimilation ability to assimilate a nucleoside. As a method for allowing a microorganism having nucleoside assimilation ability to assimilate a nucleoside in step (A), for example, a method of adding a microorganism having nucleoside assimilation ability to a wort fermentation broth and then performing a reaction by the microorganism in the wort fermentation broth can be mentioned.

[0019] In step (A), when adding a microorganism having nucleoside assimilation ability to a wort fermentation broth, the wort fermentation broth preferably contains nucleosides such as adenosine, guanosine, inosine, and xanthosine. The content of nucleosides (total content of adenosine, guanosine, inosine, and xanthosine) in the wort fermentation broth is not particularly limited, but may be, for example, 1 to 50 ppm.

[0020] In step (A), the addition amount of the above microorganism when adding a microorganism having nucleoside assimilation ability to a wort fermentation broth is not particularly limited, but with respect to the wort fermentation broth, for example, it may be 1 to 200×10 6 cells / mL, and 10 to 100×10 6 cells / mL is preferable.

[0021] The reaction conditions for allowing microorganisms capable of assimilating nucleosides to assimilate are not particularly limited as long as the microorganisms capable of assimilating nucleosides can assimilate them, and can be appropriately set depending on the type of microorganism capable of assimilating nucleosides, the amount added, the nucleoside content in the wort ferment, etc. The reaction temperature may be, for example, 5 to 35°C, and is preferably 10 to 30°C. The reaction time may be, for example, 5 hours to 10 days, and is preferably 16 hours to 5 days. To enable microorganisms capable of assimilating nucleosides to assimilate nucleosides, it is preferable to add these microorganisms to the wort fermentation liquid and then carry out the reaction under aerobic conditions. In other words, it is preferable to carry out the reaction while supplying oxygen to the reaction vessel. Alternatively, the reaction may be carried out while stirring the wort fermentation liquid to which microorganisms capable of assimilating nucleosides have been added. In the production method of the present invention, it is preferable to add microorganisms having nucleoside assimilation ability to the wort fermentation liquid, and then carry out the reaction under aerobic conditions while stirring the wort fermentation liquid to which the microorganisms having nucleoside assimilation ability have been added.

[0022] In the manufacturing method of the present invention, the addition of microorganisms having nucleoside assimilation ability in step (A) can be performed at any time during the manufacturing process of beer-flavored beverages, from the start of fermentation by brewing yeast to the filling step. For example, microorganisms having nucleoside assimilation ability can be added to the wort ferment obtained by fermentation using ordinary brewing yeast and the reaction can be continued.

[0023] The manufacturing method of the present invention does not exclude an embodiment that includes a step (B) of adding nucleosidase in addition to the above-described step (A), but from the viewpoint of reducing the number of steps and energy costs when manufacturing beer-flavored beverages, it is preferable not to include the step of adding nucleosidase (B). Furthermore, from the viewpoint of reducing the number of steps and energy costs when manufacturing beer-flavored beverages, it is preferable that the manufacturing method of the present invention does not include a heat treatment step such as boiling after fermentation with brewing yeast.

[0024] In the manufacturing method of the present invention, step (A) described above may be appropriately combined with other methods for reducing the purine content, as long as the effects of the present invention are not impaired. Methods for reducing the purine content other than steps (A) and (B) are not particularly limited, but include, for example, reducing the malt ratio, adsorption and removal of purines using adsorbents such as activated carbon, dilution, assimilation of purines by microorganisms such as yeast, and decomposition of purines using enzymes such as xanthine oxidase. Each of these methods may be combined with step (A) individually, or two or more may be combined with step (A). These methods may be performed at any timing from the mashing process to the filling process. The specific conditions for these methods are not particularly limited, and conventionally known conditions may be adopted. The adsorbents used for adsorption and removal of purines are not particularly limited, but include activated carbon, zeolite, activated clay, etc.

[0025] The following describes the beer-flavored beverage obtained by the manufacturing method of the present invention. 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, the term "beer-flavored beverage" includes not only fermented beer-flavored beverages (including beer) which are malt-fermented beverages obtained by fermenting malt, hops, and water using yeast, but also carbonated beverages to which beer flavorings, such as esters, higher alcohols, and lactones, have been added. The beer-flavored beverage obtained by the manufacturing method of the present invention is preferably 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.

[0026] The malt ratio of the beer-flavored beverage obtained by the manufacturing method of the present invention is not particularly limited and may be 25% by mass or more, 30% by mass or more, 35% by mass or more, 40% by mass or more, 45% by mass or more, 50% by mass or more, 55% by mass or more, 60% by mass or more, 65% by mass or more, 66% by mass or more, more than 66% by mass, 67% by mass or more, 70% by mass or more, 75% by mass or more, 80% by mass or more, 85% by mass or more, 90% by mass or more, 95% by mass or more, or 100% by mass. Alternatively, it may be 100% by mass or less, less than 100% by mass, 98% by mass or less, 95% by mass or less, 90% by mass or less, 87% by mass or less, 85% by mass or less, 82% by mass or less, 80% by mass or less, 78% by mass or less, 76% by mass or less, 74% by mass or less, or less than 66% by mass. 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.

[0027] 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 obtained by the manufacturing method of the present invention, it is preferable that the malt is barley malt.

[0028] The beer-flavored beverage obtained by the manufacturing method of the present invention may also use grains other than malt, protein, sugar solution, etc., as ingredients other than malt. 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.

[0029] The beer-flavored beverage obtained by the manufacturing method of the present invention has a reduced purine content compared to a beer-flavored beverage produced without going through step (A) in the manufacturing method of the present invention. In this specification, "purine" refers to a total of eight compounds, consisting of four nucleosides (adenosine, guanosine, inosine, and xanthosine) and four purine bases (adenine, guanine, hypoxanthine, and xanthine). In this specification, "purine content" refers to the total content of adenosine, guanosine, inosine, xanthosine, adenine, guanine, hypoxanthine, and xanthine. In this specification, "nucleoside content" refers to the total content of adenosine, guanosine, inosine, and xanthosine. In the beer-flavored beverage obtained by the manufacturing method of the present invention, the purine content is not particularly limited, but from a health-conscious standpoint, it is preferably 15 ppm or less, 14 ppm or less, 13 ppm or less, 12 ppm or less, 11 ppm or less, 10 ppm or less, 9 ppm or less, 8 ppm or less, 7 ppm or less, 6 ppm or less, 5 ppm or less, 4 ppm or less, 3 ppm or less, 2 ppm or less, 1.5 ppm or less, or 1 ppm or less. The lower limit of the purine content is not particularly limited, but for example, it can be 0 ppm, 0.01 ppm, or 0.1 ppm. In the beer-flavored beverage obtained by the manufacturing method of the present invention, the nucleoside content is not particularly limited, but from a health-conscious viewpoint, it is preferably 5 ppm or less, 4 ppm or less, 3 ppm or less, 2 ppm or less, 1.5 ppm or less, or 1 ppm or less. The lower limit of the nucleoside content is not particularly limited, but for example, it can be 0 ppm, 0.01 ppm, or 0.1 ppm. In this specification, the content of each of the eight types of purines in beer-flavored beverages refers to the values ​​measured using a liquid chromatography-tandem mass spectrometer (LC-MS / MS) ("Guide to 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). The purine content can be calculated by summing these measured values. In addition, the nucleoside content can be calculated by summing the measured values ​​of adenosine, guanosine, inosine, and xanthosine. The beer-flavored beverage obtained by the manufacturing method of the present invention may be a low-purine beverage, a purine-free beverage, or a purine-free beverage.

[0030] 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% by weight.

[0031] The beer-flavored beverage obtained by the manufacturing method of the present invention may be an alcohol-containing beer-flavored beverage with an alcohol content of 1 (v / v) or more, or a non-alcoholic beer-flavored beverage with an alcohol content of less than 1 (v / v). Preferably, the beer-flavored beverage obtained by the manufacturing method of the present invention is 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), or they 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] When the beer-flavored beverage obtained by the manufacturing method 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, or 4.8% by volume. Preferably, the amount is % or more, 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 obtained by the manufacturing method 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 obtained by the manufacturing method 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 and other sugary substances, salt or miso; flowers or tea, coffee, cocoa or preparations thereof; oysters, kelp, wakame seaweed or bonito flakes, etc. If the beer-flavored beverage obtained by the manufacturing method 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] When the beer-flavored beverage obtained by the manufacturing method of the present invention is to be 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.

[0052] The beer-flavored beverage obtained by the manufacturing method of the present invention may contain aliphatic alcohols from the viewpoint of alcoholic beverage characteristics. While there are no particular limitations on the aliphatic alcohols, known aliphatic alcohols with 4 to 5 carbon atoms are preferred. Examples of 4-carbon aliphatic alcohols include 2-methyl-1-propanol and 1-butanol. Examples of 5-carbon aliphatic alcohols include 3-methyl-1-butanol, 1-pentanol, and 2-pentanol. These may be used individually or in combination of two or more.

[0053] In the beer-flavored beverage obtained by the production method 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.

[0054] The beer-flavored beverage obtained by the manufacturing method 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 obtained by the manufacturing method 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 / 1 00mL 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 Bottom, 0.85g / 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 It may also be 0.55g / 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, or 0.65g / 100mL or more. It may also be 0.7g / 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 obtained by the manufacturing method of the present invention preferably has a carbohydrate content of less than 0.5 g / 100 mL.

[0055] 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.

[0056] The beer-flavored beverage obtained by the manufacturing method of the present invention typically contains peptides with a molecular weight of 35 to 50 kDa derived from malt, etc. Peptides with a molecular weight of 35 to 50 kDa are peptides found in the molecular weight range of 35 to 50 kDa, measured by ultrafiltration of the raw material liquid of the beer-flavored beverage using a 30 kDa cutoff membrane, followed by electrophoresis by SDS-PAGE. 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 obtained by the manufacturing method 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.

[0057] The beer-flavored beverage obtained by the manufacturing method of the present invention contains other peptides derived from malt, etc., in addition to the 40kDa peptide described above. Other peptides are peptides with a molecular weight of less than 35kDa or greater than 50kDa, and more specifically, peptides other than the 40kDa peptide found in the molecular weight range of 35 to 50kDa (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 obtained by the manufacturing method of the present invention, the total peptide content, which is the sum of the 40kDa peptide and other peptides, is not particularly limited, but for example, 100 to 500 ppm is preferred, and 200 to 400 ppm is more preferred. In this specification, total peptide content refers to the value measured by the Lowry method.

[0058] In the beer-flavored beverage obtained by the manufacturing method of the present invention, the total polyphenol content is not particularly limited, but from the viewpoint of drinkability, it may be, for example, 40 ppm or more, 50 ppm or more, 60 ppm or more, 70 ppm or more, 80 ppm or more, 90 ppm or more, 100 ppm or more, 110 ppm or more, or 250 ppm or less, 220 ppm or less, 200 ppm or less, 180 ppm or less, 160 ppm or less, 150 ppm or less, 140 ppm or less, 130 ppm or less, or 120 ppm or less. In this specification, 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). 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.). Furthermore, the total polyphenol content of the beer-flavored beverage obtained by the manufacturing method 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.

[0059] In the beer-flavored beverage obtained by the manufacturing method of the present invention, the total nitrogen content is not particularly limited and may be, for example, 15 mg / 100 mL or more, 20 mg / 100 mL or more, or 25 mg / 100 mL or more, or 100 mg / 100 mL or less, 95 mg / 100 mL or less, 90 mg / 100 mL or less, 85 mg / 100 mL or less, or 80 mg / 100 mL or less. In this specification, "total nitrogen content" refers to the total amount of all nitrogen compounds, including proteins and compounds containing amino acids. The total nitrogen content of the beer-flavored beverage obtained by the production method 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). The total nitrogen content of beer-flavored beverages 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 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 wort filtration time; 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; 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 beer filtration conditions.

[0060] The beer-flavored beverage obtained by the manufacturing method 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 obtained by the manufacturing method 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.

[0061] The beer-flavored beverage obtained by the manufacturing method of the present invention may be a packaged beverage. Any form or 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.

[0062] 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]

[0063] 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.

[0064] (Materials used) Commercially available beer-flavored beverage A (malt ratio: 50% by weight or more, carbohydrate content: 0.2g / 100mL, alcohol content: 5.5v / v%) The total nitrogen content of commercially available beer-flavored beverage A was measured using 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), and was found to be 18 mg / 100 mL.

[0065] <Example 1> As a microorganism possessing nucleoside assimilation ability, the yeast Lachancea thermotolerans (strain number: NBRC-1985), obtained from the National Institute of Technology and Evaluation (NITE) Biotechnology Center (NBRC), was used to obtain the sample for Example 1 by the following method.

[0066] (preculture) First, yeast pre-culture was performed. The wort used for pre-culture was prepared using 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 Lachancea thermotolerans was inoculated using one platinum loop. Pre-culture was then performed under the following conditions. Culture temperature: 30℃ Culture time: 24hr Stirring speed: 120rpm The resulting pre-culture medium of Lachancea thermotolerans was used in the following reaction.

[0067] (Nucleoside assimilation reaction conditions) Step (A): A base beer was prepared by adding glucose to a commercially available beer-flavored beverage A to a concentration of 0.5% by weight. 250 mL of the base beer was placed in a 1000 mL Erlenmeyer flask, and a pre-culture solution of Lachancea thermotolerans was added. The reaction was carried out under the following aerobic conditions. Reaction temperature: 20℃ Reaction time: 120 hours Stirring speed: 120rpm After the reaction, the reaction mixture was filtered to obtain a filtrate. The obtained filtrate was used as a sample.

[0068] <Example 2> Samples for Example 2 were obtained in the same manner as in Example 1, except that the yeast Lachancea fermentati (strain number: NBRC-479), obtained from the National Institute of Technology and Evaluation (NITE) Biotechnology Center (NBRC) as a microorganism with nucleoside assimilation ability, was used instead of the yeast Lachancea thermotolerans.

[0069] <Example 3> Samples for Example 3 were obtained in the same manner as in Example 1, except that the yeast Kluyveromyces marxianus (strain number: NBRC-10005), obtained from the National Institute of Technology and Evaluation (NITE) Biotechnology Center (NBRC) as a microorganism with nucleoside assimilation ability, was used instead of the yeast Lachancea thermotolerans.

[0070] <Example 4> Samples for Example 4 were obtained in the same manner as in Example 1, except that the yeast Zygosaccharomyces rouxii (strain number: NBRC-1130), obtained from the National Institute of Technology and Evaluation (NITE) Biotechnology Center (NBRC) as a microorganism with nucleoside assimilation ability, was used instead of the yeast Lachancea thermotolerans.

[0071] <Example 5> Samples for Example 5 were obtained in the same manner as in Example 1, except that Torulaspora delbrueckii (strain number: NBRC-955), a microorganism with nucleoside assimilation ability obtained from the National Institute of Technology and Evaluation (NITE) Biotechnology Center (NBRC), was used instead of the yeast Lachancea thermotolerans.

[0072] <Comparative Example 1> A sample for Comparative Example 1 was obtained in the same manner as in Example 1, except that the yeast Saccharomyces cerevisiae (strain number: NBRC-10217), obtained from the National Institute of Technology and Evaluation (NITE) Biotechnology Center (NBRC), was used instead of the yeast Lachancea thermotolerans.

[0073] <Comparative Example 2> A sample for Comparative Example 2 was obtained in the same manner as in Example 1, except that the yeast Saccharomyces pastrianus (strain number: NBRC-11024), obtained from the National Institute of Technology and Evaluation (NITE) Biotechnology Center (NBRC), was used instead of the yeast Lachancea thermotolerans.

[0074] (Measurement of the content of each of the 8 types of purines) The content of each of the eight types of purines in commercially available beer-flavored beverage A (Reference Example 1), as well as in each sample obtained in Examples 1-5 and Comparative Examples 1-2, was measured by the following method.

[0075] The content of adenosine, guanosine, inosine, xanthosine, adenine, guanine, hypoxanthine, and xanthine was measured using a liquid chromatography-tandem mass spectrometer (LC-MS / MS) (SCIEX X500R QTOF) under the following conditions. The purine content was calculated by summing these measurements. 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℃

[0076] Furthermore, for each sample obtained in Examples 1-5 and Comparative Examples 1-2, the nucleoside retention rate was calculated using the following formula based on the nucleoside content of commercially available beer-flavored beverage A (Reference Example 1) (total content of adenosine, guanosine, inosine, and xanthosine) and the nucleoside content of each sample. Nucleoside retention rate (%) = (Nucleoside content of each sample × 100) / Nucleoside content of commercially available beer-flavored beverage A (Reference Example 1) The results are shown in Table 1.

[0077] [Table 1]

[0078] The results in Table 1 show that the manufacturing method of the present invention can produce a beer-flavored beverage with reduced purine content by decomposing nucleosides while reducing the number of steps and energy costs.

Claims

1. A method for producing a beer-flavored beverage, comprising step (A) of adding a microorganism having nucleoside assimilation ability to a wort fermentation liquid.

2. The method for producing a product according to claim 1, wherein in step (A), a microorganism having nucleoside assimilation ability assimilates a nucleoside.

3. The method for producing a product according to claim 1 or 2, wherein the microorganism having nucleoside assimilation ability is a yeast having nucleoside assimilation ability.

4. The method for producing a product according to claim 3, wherein the yeast having nucleoside assimilation ability is at least one selected from the group consisting of yeasts belonging to the genus Lachancea, yeasts belonging to the genus Zygosaccharomyces, and yeasts belonging to the genus Kluyveromyces.

5. The method for producing the product according to claim 3, wherein the yeast having nucleoside assimilation ability is at least one selected from the group consisting of Lachancea thermotolerance, Lachancea fermentati, Zygosaccharomyces rouxii, and Kluyveromyces marxianus.

6. The manufacturing method according to claim 1 or 2, which does not include the step (B) of adding nucleosidase.

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