JP7811231B2 - Beer-flavored fermented alcoholic beverage with a satisfying taste and its manufacturing method - Google Patents

Description

The present invention relates to a beer-flavored fermented alcoholic beverage having a satisfying drinking sensation and a method for producing the same.

In response to the diversification of consumer food preferences and growing health consciousness in recent years, a wide variety of beer-flavored fermented alcoholic beverages have been developed. Because the richness of flavor that contributes to a satisfying drinking experience is an important factor that characterizes beer-flavored fermented alcoholic beverages, various techniques have been developed to improve the flavor of beer-flavored fermented alcoholic beverages. For example, by increasing the amount of malt used and reducing the final degree of fermentation, it is possible to produce a beer-flavored fermented alcoholic beverage with a rich and full-bodied flavor, while by reducing the amount of malt used and increasing the final degree of fermentation, it is possible to produce a beer-flavored fermented alcoholic beverage with a refreshing flavor.

However, increasing the flavor depth of a beer-flavored fermented alcoholic beverage also increases the intensity of the aftertaste, which lingers after consumption, thereby impairing the crispness that is valued in beer-flavored fermented alcoholic beverages. To date, a beer-flavored fermented alcoholic beverage with a refreshing taste while retaining richness and flavor depth has been developed by reducing the amount of α-glucan branching during the production process (Patent Document 1).

JP 2018-102288 A

The present invention aims to provide a novel beer-flavored fermented alcoholic beverage that is both satisfying to drink and crisp, and a method for producing the same.

The present inventors have now discovered that a beer-flavored fermented alcoholic beverage with a 2600 to 4600 D
It has been found that by adjusting the content ratio of peptide a, it is possible to suppress the aftertaste while increasing the depth of flavor. The present invention is based on this finding.

According to the present invention, the following inventions are provided.
[1] A beer-flavored fermented alcoholic beverage made at least in part from malt and/or ungerminated barley and wheat, in which the ratio (percentage) of the mass of peptides in the beverage having a molecular weight of 2600 to 4600 Da (gel filtration for HPLC analysis) to the total mass of peptides in the beverage fractionated by gel filtration for HPLC analysis is 11 to 17%.
[2] A beer-flavored fermented alcoholic beverage described in [1] above, having a carbohydrate concentration of less than 0.5 g/100 mL.
[3] A beer-flavored fermented alcoholic beverage according to [1] or [2] above, in which the malt usage ratio is 50% or more.
[4] A method for producing a beer-flavored fermented alcoholic beverage using malt and/or ungerminated barley and wheat as at least part of the raw materials, the method comprising adjusting the ratio (percentage) of the mass of peptides in the beverage having a molecular weight of 2600 to 4600 Da (gel filtration for HPLC analysis) to the total mass of peptides in the beverage fractionated by gel filtration for HPLC analysis to 11 to 17%.
[5] The production method described in [4] above, comprising a step of adding a peptide having a molecular weight of 2600 to 4600 Da (gel filtration method for HPLC analysis) contained in a beer-flavored fermented alcoholic beverage that uses malt and/or ungerminated barley as at least part of its raw materials, during the production process of the beer-flavored fermented alcoholic beverage.
[6] A flavor improver for a beer-flavored fermented alcoholic beverage, comprising as an active ingredient a peptide having a molecular weight of 2600 to 4600 Da (HPLC analytical gel filtration method) contained in a beer-flavored fermented alcoholic beverage that uses malt and/or ungerminated barley as at least part of its raw materials.
[7] A method for improving the flavor of a beer-flavored fermented alcoholic beverage using malt and/or ungerminated barley as at least a part of the raw material, comprising: determining a molecular weight of 2600 to 4600 Da (HPLC) in the beverage relative to the total peptide mass of the beverage fractionated by gel filtration for HPLC analysis;
and adjusting the mass ratio (percentage) of peptides (analytical gel filtration method) to 11-17%.

According to the present invention, a novel beer-flavored fermented alcoholic beverage that is both satisfying to drink and crisp can be provided.

FIG. 1 shows a calibration curve prepared from retention times and molecular weights of known substances in the gel filtration method for HPLC analysis carried out in Example 1.

Specific Description of the Invention

In the present invention, the term "beer taste" refers to the taste and aroma specific to beer that can be obtained when beer is produced in the usual manner, i.e., when beer is produced based on fermentation by yeast or the like.

In the present invention, a "beer-taste fermented alcoholic beverage" refers to a beer-taste fermented alcoholic beverage obtained by fermenting a carbon source, a nitrogen source, water, and the like, with yeast,
Examples include beer, happoshu, and beverages made by adding alcohol to beer or happoshu, which are made using malt as an ingredient (for example, new genre liqueur beverages classified as "liqueur (sparkling) (1)" under the Liquor Tax Act).

The beer-taste fermented alcoholic beverage of the present invention uses malt and/or ungerminated barley as part of its raw materials, and can use any malt ratio, for example, less than 50%, 50% or more, less than 67%, 67% or more, or 95% or more. In this specification, the term "malt ratio" refers to the ratio of the mass of malt to the mass of all raw materials excluding brewing water.

The beer-taste fermented alcoholic beverage of the present invention may be a beverage with a reduced carbohydrate concentration. For example, the carbohydrate concentration of the beer-taste fermented alcoholic beverage of the present invention may be 0.5 g/10
It can be less than 0 mL (equivalent to zero carbohydrates).

The carbohydrate concentration can be measured by known methods, and the moisture content (vacuum heating drying method), protein content (combustion method), lipid content (ether extraction method), and ash content (direct incineration method) can be determined from the mass of the sample.
and dietary fiber content (Prosky method (enzyme-weight method)) (see Food Labeling Standards (March 30, 2015, Dietary Fiber Table No. 139) Appendix: Analytical Methods for Nutritional Components, etc.).

In the beer-flavored fermented alcoholic beverage of the present invention, the molecular weight in the beverage is 2600 to 4600 Da (HP) relative to the total peptide mass derived from the beverage as fractionated by gel filtration for HPLC analysis.
The beverage is characterized in that the mass ratio (percentage) of peptides (HPLC gel filtration method) is within a specific range. This ratio can be calculated by dividing the concentration (mg/mL) of peptides with a molecular weight of 2600 to 4600 Da (HPLC gel filtration method) in the beverage by the total peptide concentration (mg/mL) derived from the beverage fractionated by HPLC gel filtration method. In the present invention, the lower limit of the peptide ratio (not less than or exceeding) is 1100 Da from the viewpoint of imparting depth of flavor.
% (preferably 12%), and the upper limit (not more than or less than) can be 17% (preferably 16%) from the viewpoint of suppressing aftertaste. These lower and upper limits can be arbitrarily combined, and for example, the peptide ratio range can be 1
It can be set to 1 to 17%, preferably 12 to 16%.

In the beer-flavored fermented alcoholic beverage of the present invention, the molecular weight of the beverage is 2,600 to 46
The lower limit (not less than or exceeding) of the concentration of a peptide having a molecular weight of 2600 to 4600 Da (gel filtration method for HPLC analysis) can be set to 0.15 mg/mL (preferably 0.17 mg/mL), and the upper limit (not more than or less than) can be set to 0.42 mg/mL (preferably 0.38 mg/mL). These lower and upper limits can be combined in any manner, and for example, the concentration of a peptide having a molecular weight of 2600 to 4600 Da (gel filtration method for HPLC analysis) in a beverage can be set to 0.15 to 0.42 mg/mL or 0.17 to 0.38 mg/mL.

In the beer-taste fermented alcoholic beverage of the present invention, the lower limit (greater than or equal to) of the ratio of the concentration (mg/mL) of peptides having a molecular weight of 2600 to 4600 Da (gel filtration method for HPLC analysis) in the beverage to the original extract concentration (OE concentration) (°P) in the beverage can be set to 0.018 (preferably 0.020), and the upper limit (less than or equal to) of 0.050 (preferably 0.046). These lower and upper limits can be arbitrarily combined; for example, the ratio of the concentration (mg/mL) of peptides having a molecular weight of 2600 to 4600 Da (gel filtration method for HPLC analysis) in the beverage to the OE concentration (°P) in the beverage can be set to 0.018 to 0.050 (preferably 0.020 to 0.046).
6).

The lower limit (not less than or exceeding) of the OE concentration in the beer-taste fermented alcoholic beverage of the present invention can be 2°P (preferably 4°P), and the upper limit (not more than or below) can be 20°P (preferably 14°P, more preferably 10°P). These lower and upper limits can be combined in any desired manner. For example, the OE concentration in the beer-taste fermented alcoholic beverage of the present invention can be set to 2 to 20°P (preferably 4 to 14°P).
In the present invention, the "original extract concentration" can be measured using a commercially available device (for example, an Alcolyzer (manufactured by Anton Paar)). In the case of a beer-flavored fermented alcoholic beverage in which malt and/or ungerminated barley or wheat is used as at least a part of the raw materials, the "original extract concentration" can be rephrased as the "original wort extract concentration."

The molecular weight of peptides in beverages is measured by gel filtration using high performance liquid chromatography (sometimes referred to herein as "gel filtration for HPLC analysis"). Specifically, the molecular weight is calculated from the retention time using a calibration curve for gel filtration for HPLC analysis (e.g., Figure 1), and samples with different molecular weights can be separated based on the retention time. A specific example of molecular weight measurement using gel filtration for HPLC analysis is shown in Example 1 below. Alternatively, peptide quantification can be performed by the Lowry method.

The beer-taste fermented alcoholic beverage of the present invention is characterized by enhanced depth of flavor while suppressing aftertaste. Here, "depth of flavor" refers to the flavor sensation recognized by the breadth of flavor, complexity, and full-bodiedness, and "aftertaste" refers to the flavor sensation consisting of the five basic tastes and stimulating flavors (sweetness, sourness, saltiness, umami, bitterness, astringency, spiciness, roughness, etc.) that remain in the oral cavity after swallowing. In a beer-taste fermented alcoholic beverage, depth of flavor contributes to its satisfying drinking experience, and suppressing the aftertaste increases its crispness. Therefore, the present invention can provide a beer-taste fermented alcoholic beverage that is both satisfying and crisp.

The beer-taste fermented alcoholic beverage of the present invention can be produced according to a normal production procedure for a beer-taste fermented alcoholic beverage, as long as the ratio of 2600 to 4600 Da peptides in the beverage is adjusted to fall within a predetermined range.

For example, a beer-flavored fermented alcoholic beverage can be produced by adding fermentation beer yeast to wort prepared from brewing raw materials such as malt, hops, auxiliary materials, and brewing water, fermenting the wort, storing the resulting fermented liquid at low temperature, and then removing the yeast through a filtration process.
Among beer-flavored fermented alcoholic beverages, all-malt beer is made from malt, hops,
It goes without saying that it can be produced from water.

In the above-mentioned production procedure, wort can be produced by a conventional method. For example, a mixture of brewing raw materials and brewing water is saccharified, filtered to obtain wort, hops are added to the wort, the wort is boiled, and the boiled wort is cooled to prepare the wort.
Commercially available enzyme preparations can also be added during the saccharification process. For example, protease preparations can be used for protein degradation, α-amylase preparations, glucoamylase preparations, pullulanase preparations, etc. for carbohydrate degradation, and β-glucanase preparations, cellulose-degrading enzyme preparations, etc. for cellulose degradation, or a mixture of these preparations can also be used.

In one embodiment of the production of the beer-taste fermented alcoholic beverage of the present invention, malt with high endoprotease activity and/or an enzyme preparation possessing endoprotease activity can be used for proteolysis in the saccharification step or fermentation step. By carrying out such steps, it is possible to obtain the 26% molecular weight malt protein contained in the beer-taste fermented alcoholic beverage.
By increasing the ratio of peptides in the range of 00 to 4600 Da (gel filtration method for HPLC analysis) and adjusting it to fall within a predetermined range, it is possible to produce a beer-taste fermented alcoholic beverage with enhanced flavor and reduced aftertaste. That is, the present invention provides a method for producing a beer-taste beverage, which comprises using malt with high endoprotease activity and/or an enzyme preparation containing endoprotease in the saccharification step or fermentation step. The characteristics of the enzymatic activity of the malt or enzyme preparation can be identified based on the endoprotease activity.

The beer-taste fermented alcoholic beverage of the present invention uses either malt or ungerminated barley or wheat as at least a part of the raw materials, and preferably uses malt or malt and ungerminated barley or wheat as at least a part of the raw materials. In the beer-taste fermented alcoholic beverage of the present invention, barley malt and/or wheat malt can be used as the malt as a part of the raw materials. In the beer-taste fermented alcoholic beverage of the present invention,
As the ungerminated wheat and barley, ungerminated barley (including extract) and ungerminated wheat (including extract) can be used as raw materials.

In the production of the beer-taste fermented alcoholic beverage of the present invention, in addition to malt and ungerminated wheat and barley, rice, corn, soybeans, koryuan (rice bran), potatoes, starch, sugars (e.g., liquid sugar),
Fruits (e.g., juice, juice concentrate), coriander or its seeds, spices or their ingredients (
secondary ingredients such as pepper, cinnamon, Japanese pepper), herbs (e.g., chamomile, sage, basil), vegetables (e.g., sweet potato, pumpkin), buckwheat or sesame, sugar-containing substances (e.g., honey, brown sugar), salt, miso, flowers, tea, coffee, cocoa (tea, coffee, and cocoa include preparations thereof), seafood (e.g., oysters, kelp, wakame seaweed, bonito flakes);
Nitrogen sources such as protein hydrolysates and yeast extract, as well as other additives such as flavorings, colorants, foaming/foam retention improvers, water quality conditioners, and fermentation aids can be used as brewing ingredients. The raw materials used in the beer-taste fermented alcoholic beverage of the present invention, 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.

In the present invention, a fraction containing peptides with a molecular weight of 2600 to 4600 Da is prepared from grains (e.g., malt and/or ungerminated wheat or soybeans, preferably barley malt or wheat malt), and the fraction is added to a beer-taste fermented alcoholic beverage, thereby adjusting the proportion of peptides with a molecular weight of 2600 to 4600 Da to fall within a predetermined range,
In other words, according to the present invention, a beer-flavored fermented alcoholic beverage having a reduced aftertaste and an enhanced flavor can be produced.
The present invention provides a method for producing a beer-taste fermented alcoholic beverage, comprising preparing a fraction containing a peptide of molecular weight 2600 to 4600 Da (gel filtration method for HPLC analysis), and adding the fraction to a beer-taste fermented alcoholic beverage. The peptide fraction can be added to the beer-taste fermented alcoholic beverage during the production process of the beverage (e.g., blending the peptide into a fermentation broth). Cereal-derived peptides with a molecular weight of 2600 to 4600 Da (gel filtration method for HPLC analysis) can be prepared, for example, by hydrolyzing cereals with a protease such as an endoprotease, and may also be obtained from cereal protein hydrolysates such as soy protein hydrolysates.

In the present invention, a beer-taste fermented alcoholic beverage is produced using malt and/or ungerminated barley as at least a part of the raw material, and a fermented alcoholic beverage having a molecular weight of 2600 to 4600 D is obtained from the beverage.
By preparing a fraction containing peptides of formula (a) and adding this fraction to a beer-taste fermented alcoholic beverage, it is possible to adjust the ratio of peptides with a molecular weight of 2600 to 4600 Da to within a predetermined range, thereby producing a beer-taste fermented alcoholic beverage with an enhanced flavor and a reduced aftertaste. That is, according to the present invention, there is provided a method for producing a beer-taste fermented alcoholic beverage, which comprises producing a beer-taste fermented alcoholic beverage using malt and/or ungerminated barley and wheat as at least a part of the ingredients, then preparing a fraction from the beverage containing peptides with a molecular weight of 2600 to 4600 Da (gel filtration method for HPLC analysis), and adding this fraction to the beer-taste fermented alcoholic beverage.
The peptide fraction is added to a beer-flavored fermented alcoholic beverage during the production process of the beverage (
For example, this can be carried out during the preparation of the fermentation liquid.

Furthermore, according to the present invention, there is provided a method for producing a fermented alcoholic beverage with a molecular weight of 2600 to 460, which is derived from a beer-flavored fermented alcoholic beverage containing, as at least a part of the raw materials, grains or malt and/or ungerminated wheat and barley.
A beer-flavored fermented alcoholic beverage containing one or more 0 Da peptides is provided, and this beverage is a part of the beer-flavored fermented alcoholic beverage of the present invention. The beverage containing the peptide has an improved or enhanced flavor as a beer-flavored fermented alcoholic beverage, specifically, a beverage with enhanced flavor while suppressing the aftertaste.

As described above, the beer-taste fermented alcoholic beverage of the present invention can be provided in a form in which the carbohydrate content has been reduced. The carbohydrate content can be reduced by known methods, such as a method in which glucoamylase is added during the saccharification process, a method in which glucoamylase is added during the fermentation process (
Enzyme Utilization Technology Series: From Basics and Analysis to Modification, Advanced Functionalization, and Industrial Use (Edited by Makoto Komiyama,
NTS Corporation, 2010), etc.), a method of reducing carbohydrates by increasing the assimilation rate of yeast using liquid sugar containing a large amount of carbohydrates that can be assimilated by yeast (see JP 2009-
131202), a method of filtering wort during the saccharification process (see JP 2012-1477
By using this method (see Patent Publication No. 80), the carbohydrate concentration in the beverage can be adjusted to a predetermined value.

The beverage provided by the present invention can be provided as a bottled beverage after being subjected to a step of adding carbon dioxide gas, and further subjected to steps such as a filling step and a sterilization step. Sterilization may be performed before or after filling into the container. Furthermore, if the pH of the beverage is adjusted to less than 4, it can be filled directly to produce a bottled beverage without undergoing the sterilization step.

The container used for the beverage of the present invention may be any container that is commonly used for filling beverages, such as a metal can, a barrel, a plastic bottle (e.g., a PET bottle, a cup),
Examples of the container include paper containers, bottles, and pouch containers, but metal cans, barrels, plastic bottles (for example, PET bottles), and bottles are preferred.

According to another aspect of the present invention, there is provided a method for producing a beer-flavored fermented alcoholic beverage having an improved flavor, using malt and/or ungerminated barley as at least a part of the raw materials, the method comprising:
and adjusting the ratio (percentage) of the mass of peptides in the beverage having a molecular weight of 2600 to 4600 Da (gel filtration for HPLC analysis) to the total peptide mass of the beverage fractionated by gel filtration for LC analysis to 11 to 17%. In the present invention, "improved flavor" or "flavor improvement" of a beer-taste fermented alcoholic beverage refers to enhancing the depth of flavor while suppressing the aftertaste of the beer-taste fermented alcoholic beverage. The above-mentioned production method of the present invention is preferably a production method of a beer-taste fermented alcoholic beverage that enhances the depth of flavor while suppressing the aftertaste. The above-mentioned production method of the present invention can be carried out according to the description of the beer-taste fermented alcoholic beverage and its production method of the present invention.

According to another aspect of the present invention, there is provided a method for producing a fermented alcoholic beverage with a beer taste, the method comprising the steps of: (a) providing a fermented alcoholic beverage containing grains or malt and/or ungerminated wheat and barley as at least a part of the raw material;
The present invention provides a flavor-improving agent for a beer-taste fermented alcoholic beverage, comprising a peptide of 0.00 Da (gel filtration method for HPLC analysis) as an active ingredient. The flavor-improving agent for a beer-taste fermented alcoholic beverage is preferably an agent for enhancing the flavor depth or drinking response of a beer-taste fermented alcoholic beverage. The flavor-improving agent of the present invention can be produced according to the description of the beer-taste fermented alcoholic beverage and the method for producing the same of the present invention.

According to yet another aspect of the present invention, there is provided a method for improving the flavor of a beer-taste fermented alcoholic beverage that uses malt and/or ungerminated barley as part of its ingredients. The flavor improvement method of the present invention can be carried out by adjusting the ratio (percentage) of the mass of peptides in the beverage having a molecular weight of 2600 to 4600 Da (gel filtration for HPLC analysis) to the total peptide mass of the beverage fractionated by gel filtration for HPLC analysis to 11 to 17%. The flavor improvement method of the present invention is preferably a method for enhancing the depth of flavor of a beer-taste fermented alcoholic beverage or a method for enhancing the drinking experience of the beverage. The flavor improvement method of the present invention can be carried out according to the description of the beer-taste fermented alcoholic beverage of the present invention and the method for producing the same.

The present invention will be described in more detail based on the following examples, but the present invention is not limited to these examples.

Example 1: Production of a beer-flavored fermented alcoholic beverage and the effect of adding a peptide to the beverage
Analysis of the effect on the thickness and aftertaste of the beer-flavored fermented alcoholic beverages (Beer Bases 1 and 2) A. Production of Beer Base 1 In the production of the beer-flavored fermented alcoholic beverage of this example, barley malt was used as the main raw material. An enzyme preparation was used for saccharification, and the saccharification temperature and time were adjusted, followed by filtration to obtain wort. Specifically, 20.0 parts by mass of barley malt was added to 100 parts by mass of hot water at 62°C.
After adding the enzyme preparation, the mixture was kept for 40 minutes, heated to 78°C and kept for 5 minutes, and then filtered to obtain wort.

Following the wort preparation step, 19.0 parts by mass of hops and liquid sugar were added to the resulting wort and boiled at 100°C for 90 minutes. The wort was then allowed to stand, the trub was separated, and the mixture was cooled to obtain a pre-fermentation liquid. Bottom-fermenting yeast was then added to the pre-fermentation liquid, and main fermentation and post-fermentation were carried out according to conventional methods. The fermented liquid after post-fermentation was then stored at a lower temperature and filtered to obtain a clear beer-flavored alcoholic beverage (malt usage ratio 50%) (beer base 1) with a carbohydrate concentration of 3.1 g/100 mL.

B. Production of Beer Base 2 In the production of the beer-flavored fermented alcoholic beverage of this example, barley malt was used as the main raw material. An enzyme preparation was used for saccharification, and the saccharification temperature and time were adjusted, followed by filtration to obtain wort. Specifically, 100 parts by mass of hot water at 64°C was mixed with 10.0 parts by mass of barley malt.
Parts by weight of a β-glucanase enzyme preparation and an enzyme preparation mainly containing glucoamylase were added and kept for 100 minutes, then the temperature was raised to 78°C and kept for 5 minutes, and then filtered to obtain wort.

Following the wort preparation step, 9.5 parts by mass of liquid sugar mainly composed of hops and assimilable sugars was added to the resulting wort and boiled at 100°C for 90 minutes. The wort was then allowed to stand, the trub was separated, and the mixture was cooled to obtain a pre-fermentation liquid. Bottom-fermenting yeast was then added to the pre-fermentation liquid, and main fermentation and post-fermentation were carried out according to conventional methods. The fermented liquid after post-fermentation was then stored at a lower temperature and filtered to obtain a clear beer-flavored alcoholic beverage (malt usage ratio 50%) (beer base 2) with a carbohydrate concentration of 0.3 g/100 mL.

(2) Preparation of peptide fractions A commercially available beer-flavored beverage was degassed, weighed, and freeze-dried.
The sample was subjected to gel filtration fractionation under the following conditions:

<Gel filtration fractionation conditions>
Column: Hiload Superdex 30pg 26/600 (GE Healthcare)
Sample injection volume: 5 mL
Eluent composition: 100mM NaCl
Flow rate: 2.5mL/min (constant flow rate)
Detection wavelength: 215 nm
Fractionation: 19.1 mL (fraction 0) from 0.29 cv (column volume), then 5 mL fractions from 0.35 cv (fractions 1 to 51)

The obtained fractions (fraction numbers: F10 to F11) were purified and quantified under the following conditions.

The obtained fraction was extracted with a solid-phase extraction column (Bond Elute C18, Agilent Technology).
The peptide fraction was obtained by adsorption using a 50% ethanol solution (Gies), washed with demineralized water, and then eluted with 50% ethanol. The eluate was concentrated to dryness and reconstituted with demineralized water to obtain a concentrate. This was used as the peptide fraction.

Protein quantification of the peptide fraction obtained by purification was carried out using a commercially available kit (
The DC protein assay was performed by the Lowry method using a DC protein assay (Bio-Rad).
The concentration of the fraction solution was adjusted to an appropriate range. 50 μL of solution A was added to 5 μL of the adjusted concentration sample and stirred, followed by 400 μL of solution B and stirring. After a 15-minute color reaction at room temperature, 350 μL was transferred to a 96-well plate and the absorbance at 750 nm was measured. The peptide concentration (mg/mL) was calculated based on the obtained absorbance and a previously prepared calibration curve. The calibration curve was prepared using BSA (bovine serum albumin).

(3) Preparation of gel filtration fractions for HPLC analysis The test samples obtained in (1) above were degassed, weighed, and freeze-dried. Each freeze-dried product was dissolved in 50 mM sodium phosphate buffer (pH 7.0, 150 mM N
The peptide fraction obtained in (2) above was dissolved in the above buffer solution to a concentration equivalent to 10 times the concentration of the solution, and this was used as the sample for gel filtration fractionation. Each fraction sample was subjected to gel filtration fractionation under the following conditions to obtain fractions (fractions 1 to 10).

The conditions for gel filtration for HPLC analysis were as follows:
<Gel filtration analysis conditions for HPLC analysis>
Column: Superdex 75 10/300 (GE Healthcare)
Sample injection volume: 100 μL Eluent composition: 50 mM sodium phosphate (pH 7.0), 2
0% (v/v) acetonitrile, 150 mM NaCl
Flow rate: 0.5mL/min (constant flow rate)
Detection wavelength: 215 nm

Fractionation program:

Protein quantification of the fractions was performed using the Lowry method described above in (2).

(4) Measurement of Molecular Weight by Gel Filtration for HPLC Analysis A. Preparation of Samples for Gel Filtration for HPLC Analysis The fractions (fractions 1 to 10) obtained in (3) above were used as samples.

Using the column, eluent, flow rate, and detection wavelength described above as gel filtration conditions for HPLC analysis, peptides with known molecular weights were dissolved in ultrapure water at 0.1 to 5 mg/mL, and 50 μL of the solution was injected for HPLC analysis, and the retention time was confirmed (Table 2). A calibration curve (Figure 1) was created from the retention time and molecular weight.

C. Calculation of Molecular Weight As a result of molecular weight measurement, it was confirmed that the peptide contained in fraction 5 of fractions 1 to 10 had a molecular weight in the range of 2600 to 4600 Da.

(5) Calculation of the ratio of peptides with molecular weights of 2600 to 4600 Da The ratio of peptides with molecular weights of 2600 to 4600 Da was calculated using the peptide concentration in each fraction equivalent to the product using the following calculation formula.

(6) Analysis of Carbohydrate Concentration and Original Extract Concentration The carbohydrate concentration of the test product was measured in accordance with the method described in the 5th edition of the Japanese Standard Tables of Food Composition Analysis Manual, based on the nutrition labeling standard.
Decompression heating drying method), protein (method using an automatic analyzer), lipids (Soxhlet extraction method (3)), ash (muffle furnace combustion method), and dietary fiber content (Prosky enzyme weight method)
The original extract concentration (OE concentration) of the brewed product was measured in accordance with the BCOJ Beer Analysis Method (Beer Brewers Association (2013) 8.3.6, 8.4.3 and 8.5). The measurement was performed using an Alcolyzer (manufactured by Anton Paar).

(7) Sensory Evaluation Beer-flavored fermented alcoholic beverages (Beer Bases 1 and 2) produced in (1) above
A test group was prepared in which the peptide fraction obtained in (2) above was not added, and a test group in which it was added.
The peptide concentrations of 600 to 4600 Da were adjusted as shown in Tables 3 and 4.
Samples Nos. 1 to 10 were prepared. Test Example 1 was an example in which the peptide fraction was added to beer base 1, and Test Example 2 was an example in which the peptide fraction was added to beer base 2.

The sensory evaluation was conducted by four trained panelists. Specifically, "depth of flavor" and "aftertaste" were evaluated on a nine-point scale from 1 to 9, with increments of one point, and the average scores of the four panelists were calculated. Here, "depth of flavor" refers to the flavor sensation recognized by the breadth of flavor, complexity, and body. "Aftertaste" refers to the flavor sensation consisting of the five basic tastes and stimulating tastes (sweetness, sourness, saltiness, umami, bitterness, astringency, spiciness, roughness, etc.) remaining in the oral cavity after swallowing.

Test Example 1 (Tests 1 to 5) Evaluating the Effect of Adding Peptide Fractions to Beer Base 1
In Test Example 2 (Tests 6 to 10), which evaluated the effect of adding a peptide fraction to beer base 2, the scores for "depth of flavor" and "aftertaste" of Test Plot 1 were fixed at 3 in advance and the evaluation was carried out. In Test Example 2 (Test Plots 6 to 10), which evaluated the effect of adding a peptide fraction to beer base 2, the scores for "depth of flavor" and "aftertaste" of Test Plot 6 were fixed at 3 in advance and the evaluation was carried out. In all test examples, the depth of flavor was rated at 3.5 points or more, and the aftertaste was rated at 5.0 points or more.
The following was determined to be a preferred embodiment.

The results of the sensory evaluation are shown in Table 3.

From the results in Tables 3 and 4, it can be seen that the beer-flavored fermented alcoholic beverage (malt content: 50%)
It was confirmed that by adjusting the ratio of 2600 to 4600 Da peptides within a predetermined range, it is possible to suppress the aftertaste while enhancing the richness of the flavor. Furthermore, it was confirmed that this effect is achieved regardless of the carbohydrate concentration of the beer-flavored fermented alcoholic beverage.

In addition, the above-mentioned sensory evaluation test was conducted on Beer Bases 1 and 2 (however, the scores for "depth of flavor" and "aftertaste" were not fixed at 3 in advance), and the following results were obtained.

The results in Table 4 confirm that zero-sugar beer base 2 has a relatively less rich flavor and aftertaste compared to beer base 1. Combining the results in Tables 3 and 4 with the results in Table 5, it was confirmed that in zero-sugar beer-flavored fermented alcoholic beverages, which tend to have a reduced rich flavor, adjusting the ratio of 2600 to 4600 Da peptides to fall within a specified range can increase the richness of flavor while suppressing the aftertaste.

Claims (8)

A beer-flavored fermented alcoholic beverage made at least in part from malt, wherein the ratio of the concentration (mg/mL) of peptides in the beverage having a molecular weight of 2600 to 4600 Da (gel filtration for HPLC analysis) to the original extract concentration (OE concentration) (°P) in the beverage is 0.018 to 0.050, the ratio (percentage) of the mass of the peptides in the beverage having a molecular weight of 2600 to 4600 Da (gel filtration for HPLC analysis) to the total peptide mass of the beverage fractionated by gel filtration for HPLC analysis is 11 to 17%, the concentration of the peptides in the beverage having a molecular weight of 2600 to 4600 Da (gel filtration for HPLC analysis) is 0.15 to 0.42 mg/mL, and the peptides having a molecular weight of 2600 to 4600 Da (gel filtration for HPLC analysis) are derived from a beer-flavored fermented alcoholic beverage made at least in part from malt. The beer-flavored fermented alcoholic beverage according to claim 1 , wherein the carbohydrate concentration is less than 0.5 g/100 mL. 3. The beer-taste fermented alcoholic beverage according to claim 1, wherein the malt content is 50% or more. 3. The beer-taste fermented alcoholic beverage according to claim 1 , wherein the malt content is less than 50%. 5. The beer-flavored fermented alcoholic beverage according to claim 1, further comprising one or more secondary ingredients selected from the group consisting of ungerminated wheat, rice, corn, soybeans, koryang (dried radish), potatoes, starch, sugars, fruit, coriander or its seeds, spices or raw materials thereof, herbs, vegetables, buckwheat or sesame, carbohydrate-containing substances, salt, miso, flowers, tea, coffee, cocoa, and seafood. A method for producing a beer-taste fermented alcoholic beverage using malt as at least a part of an ingredient, the method comprising: adjusting the ratio (mg/mL) of the concentration of peptides in the beverage having a molecular weight of 2600 to 4600 Da (gel filtration for HPLC analysis) to the original extract concentration (OE concentration) (°P) in the beverage to 0.018 to 0.050; adjusting the ratio (percentage) of the mass of the peptides in the beverage having a molecular weight of 2600 to 4600 Da (gel filtration for HPLC analysis) to the total peptide mass of the beverage fractionated by gel filtration for HPLC analysis to 11 to 17%; and adjusting the concentration of the peptides in the beverage having a molecular weight of 2600 to 4600 Da (gel filtration for HPLC analysis) to 0.15 to 0.42 mg/mL , wherein the peptides are derived from a beer-taste fermented alcoholic beverage using malt as at least a part of an ingredient. The production method according to claim 6, comprising a step of adding a peptide having a molecular weight of 2600 to 4600 Da (as determined by gel filtration for HPLC analysis) contained in a beer-taste fermented alcoholic beverage that uses malt as at least a part of its raw materials, in the production process of the beer - taste fermented alcoholic beverage. A method for improving the flavor of a beer-taste fermented alcoholic beverage that uses malt as at least a part of an ingredient, the method comprising: adjusting the ratio (mg/mL) of the concentration of peptides in the beverage having a molecular weight of 2600 to 4600 Da (gel filtration for HPLC analysis) to the original extract concentration (OE concentration) (°P) in the beverage to 0.018 to 0.050; adjusting the ratio (percentage) of the mass of the peptides in the beverage having a molecular weight of 2600 to 4600 Da (gel filtration for HPLC analysis) to the total peptide mass of the beverage fractionated by gel filtration for HPLC analysis to 11 to 17%; and adjusting the concentration of the peptides having a molecular weight of 2600 to 4600 Da (gel filtration for HPLC analysis) in the beverage to 0.15 to 0.42 mg/mL , wherein the peptides are derived from a beer-taste fermented alcoholic beverage that uses malt as at least a part of an ingredient.