JP7460720B2 - Beer-taste fermented alcoholic beverage with a satisfying drinking sensation and method for producing the same - Google Patents

Description

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

A wide variety of beer-taste fermented alcoholic beverages have been developed in response to the diversification of consumer food preferences and increased health consciousness in recent years. Since the depth of flavor that contributes to the drinkability is an important element that characterizes beer-taste fermented alcoholic beverages, various techniques have been developed to improve the flavor of beer-taste fermented alcoholic beverages. For example, by increasing the amount of malt used and reducing the final fermentation degree, it is possible to produce a beer-taste fermented alcoholic beverage that has a richer and richer taste. By increasing this amount, a beer-like fermented alcoholic beverage with a refreshing taste can be produced.

However, increasing the thickness of the taste in beer-taste fermented alcoholic beverages also increases the intensity of the aftertaste, which lingers after drinking, which impairs the sharpness that is important in beer-taste fermented alcoholic beverages. To date, by reducing the amount of α-glucan branching during the manufacturing process, beer-taste fermented alcoholic beverages have been developed that have richness and depth of flavor and have a refreshing taste (Patent Document 1).

JP 2018-102288 A

An object of the present invention is to provide a novel beer-taste fermented alcoholic beverage that is both satisfying and sharp, 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-taste fermented alcoholic beverage containing malt and/or ungerminated barley as at least a part of the raw material, the beverage relative to the total peptide mass of the beverage fractionated by gel filtration for HPLC analysis. A beer-taste fermented alcoholic beverage in which the mass ratio (percentage) of peptides with a molecular weight of 2,600 to 4,600 Da (gel filtration method for HPLC analysis) is 11 to 17%.
[2] The beer-taste fermented alcoholic beverage according to [1] above, which has a carbohydrate concentration of less than 0.5 g/100 mL.
[3] The beer-taste fermented alcoholic beverage according to [1] or [2] above, wherein the malt usage ratio is 50% or more.
[4] A method for producing a beer-taste fermented alcoholic beverage using malt and/or ungerminated barley as at least a part of the raw materials, the total peptide mass of the beverage being fractionated by gel filtration for HPLC analysis. A method comprising adjusting the mass ratio (percentage) of peptides having a molecular weight of 2,600 to 4,600 Da (gel filtration method for HPLC analysis) in the beverage to 11 to 17%.
[5] In the manufacturing process of beer-taste fermented alcoholic beverages, molecular weights of 2600 to 4600 Da (gel filtration method for HPLC analysis) contained in beer-taste fermented alcoholic beverages containing malt and/or ungerminated barley as at least part of the raw materials. The manufacturing method according to [4] above, which includes the step of adding a peptide.
[6] A beer-taste fermented alcoholic beverage containing at least a part of malt and/or ungerminated barley as an active ingredient, which contains a peptide with a molecular weight of 2,600 to 4,600 Da (gel filtration method for HPLC analysis) as an active ingredient. Flavor improver for beer-taste fermented alcoholic beverages.
[7] A method for improving the flavor of a beer-taste fermented alcoholic beverage that uses malt and/or ungerminated barley as at least a part of the raw materials, the total peptides of the beverage being fractionated by a gel filtration method for HPLC analysis. Molecular weight in the beverage relative to mass: 2600 to 4600 Da (HPLC
A method comprising adjusting the mass ratio (percentage) of peptides (analytical gel filtration method) to 11-17%.

According to the present invention, it is possible to provide a novel beer-taste fermented alcoholic beverage that is both satisfying and sharp.

FIG. 2 is a diagram showing a calibration curve created from the retention time of gel filtration method for HPLC analysis and the molecular weight of a known substance, which was carried out in Example 1.

Specific description of the invention

In the present invention, "beer taste" refers to the taste and aroma unique to beer that is obtained when beer is normally produced, that is, when beer is produced based on fermentation using yeast or the like.

In the present invention, a "fermented alcoholic beverage with a beer taste" is a fermented alcoholic beverage with a beer taste that is fermented by yeast using a carbon source, a nitrogen source, water, and the like as ingredients,
Examples of such beverages include beer, happoshu, and beverages made by adding alcohol to beer or happoshu that use malt as an ingredient (for example, new liqueur-type beverages classified as "liqueur (sparkling) (1)" under the Liquor Tax Law).

The beer-taste fermented alcoholic beverage of the present invention uses malt and/or ungerminated barley as a part of the raw materials, and any malt usage ratio can be used, but for example, the malt usage ratio can be 50%. %, 50% or more, less than 67%, 67% or more, or 95% or more. As used herein, "malt usage ratio" refers to the ratio of malt mass to the mass of all raw materials excluding brewing water.

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

The carbohydrate concentration can be measured by known methods. From the mass of the sample, the moisture content (vacuum heating drying method), protein content (combustion method), lipid content (ether extraction method), ash content (direct ashing method), etc. can be calculated.
This can be done according to the method for calculating the amount of dietary fiber excluding the Prosky method (enzymatic - gravimetric method) (see the Food Labeling Standards (March 30, 2015, Digestive Health 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 ratio (percentage) of the mass of peptides (HPLC analysis gel filtration method) is within a specific range. The ratio can be calculated by dividing the concentration (mg/mL) of peptides with a molecular weight of 2600 to 4600 Da (HPLC analysis gel filtration method) in the beverage by the total peptide concentration (mg/mL) derived from the beverage fractionated by HPLC analysis gel filtration method. In the present invention, the lower limit of the peptide ratio (not less than or exceeding) is 11 to 150 mg/mL 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 is 1
It may be 1 to 17%, preferably 12 to 16%.

In the fermented beer-flavored 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 peptide concentration of 0.00 Da (gel filtration method for HPLC analysis) can be 0.15 mg/mL (preferably 0.17 mg/mL) and the upper limit (not more than or less than) can be 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 in the beverage with a molecular weight of 2600 to 4600 Da (gel filtration method for HPLC analysis) can be 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 concentration (mg/mL) of peptides with a molecular weight of 2600 to 4600 Da (gel filtration method for HPLC analysis) in the beverage is determined relative 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) 0.050 (preferably 0.046). be able to. These lower and upper limits can be arbitrarily combined, for example, the concentration (mg/ mL) to 0.018 to 0.050 (preferably 0.020 to 0.04)
6).

The lower limit (more than or above) 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 (below or less than) 20°P. (preferably 14°P, more preferably 10°P). These lower and upper limits can be arbitrarily combined, for example, the OE concentration in the beer-taste fermented alcoholic beverage of the present invention is set to 2 to 20°P (preferably 4 to 14°P).
It can be done. In the present invention, the "original extract concentration" can be measured using a commercially available device (for example, Alcolyzer (manufactured by Anton Paar)). In addition, in the case of a beer-taste fermented alcoholic beverage that uses malt and/or ungerminated barley 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 the peptide in the beverage is measured by a gel filtration method using high performance liquid chromatography (herein sometimes referred to as "gel filtration method for HPLC analysis"). Specifically, the molecular weight can be calculated from the retention time using a calibration curve of the gel filtration method for HPLC analysis (for example, FIG. 1), and samples with different molecular weights can be fractionated depending on the retention time. A specific example of molecular weight measurement by gel filtration method for HPLC analysis is as shown in Example 1 below. Moreover, quantification of peptides can be carried out by the Lowry method.

The beer-taste fermented alcoholic beverage of the present invention is characterized by having an enhanced taste while suppressing aftertaste. Here, "thickness of taste" refers to the sense of flavor perceived by the breadth of taste, complexity, body, etc., and "aftertaste" refers to the basic five tastes and stimulating tastes that remain in the mouth after swallowing. It refers to the sense of flavor consisting of sweetness, sourness, saltiness, umami, bitterness, astringency, spiciness, roughness, etc. In a beer-taste fermented alcoholic beverage, the thickness of the taste contributes to the drinkability, and suppressing the aftertaste increases the sharpness.According to the present invention, it is an object of the present invention to provide a beer-taste fermented alcoholic beverage that has both the drinkability and the sharpness. Can be done.

The fermented, beer-taste alcoholic beverage of the present invention can be produced in accordance with normal production procedures for fermented, beer-taste alcoholic beverages, so long as the ratio of 2,600 to 4,600 Da peptides in the beverage is adjusted to fall within a specified range.

For example, brewer's yeast for fermentation is added to wort prepared from brewing raw materials such as malt, hops, auxiliary raw materials, and brewing water, fermentation is performed, and the resulting fermented liquid is stored at a low temperature and then subjected to a filtration process. By removing yeast, a beer-taste fermented alcoholic beverage can be produced.
In addition, among beer-taste fermented alcoholic beverages, all-malt beer is made with malt, hops,
Needless to say, it can be produced from water.

In the above manufacturing procedure, wort can be prepared according to a conventional method. For example, wort is prepared by saccharifying and filtering a mixture of brewing raw materials and brewing water to obtain wort, adding hops to the wort, boiling it, and cooling the boiled wort. Can be done. In addition, the wort is
It can also be produced by adding a commercially available enzyme preparation during the saccharification process. For example, protease preparations are used for proteolysis, α-amylase preparations, glucoamylase preparations, pullulanase preparations, etc. are used for carbohydrate decomposition, and β-glucanase preparations, fibrinolytic enzyme preparations, etc. are used for fibrinolysis. Alternatively, a mixed formulation of these can be used.

In one embodiment of the production of the beer-taste fermented alcoholic beverage of the present invention, malt with high endo-protease activity and/or an enzyme preparation possessing endo-protease activity are used for proteolysis in the saccharification step or fermentation step. be able to. By carrying out such a process, the molecular weight contained in the beer-taste fermented alcoholic beverage is 26.
By increasing the ratio of peptides ranging from 00 to 4,600 Da (gel filtration method for HPLC analysis), it is possible to adjust the ratio to within a predetermined value range, thereby producing a beer-taste fermented alcoholic beverage that suppresses the aftertaste and enhances the depth of the taste. can be manufactured. That is, the present invention provides a method for producing a beer-taste beverage, which comprises using malt with high endo-protease activity and/or an enzyme preparation containing endo-protease in the saccharification or fermentation step. The enzyme activity characteristics of the malt and enzyme preparations described above can be specified based on the endo-protease activity.

The beer-taste fermented alcoholic beverage of the present invention uses either or both of malt and ungerminated barley as at least a part of the raw material, and preferably malt or malt and ungerminated barley as at least a part of the raw material. It shall be a part of it. In the beer-taste fermented alcoholic beverage of the present invention, barley malt and/or wheat malt can be used as part of the raw material. In the beer-taste fermented alcoholic beverage of the present invention,
As ungerminated barley, ungerminated barley (including extracts) and ungerminated wheat (including extracts) 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 barley, rice, corn, soybeans, koryan, potatoes, starch, sugars (for example, liquid sugar),
Fruits (e.g. fruit juice, fruit juice concentrate), coriander or its seeds, spices or their raw materials (
(e.g. pepper, cinnamon, Japanese pepper), herbs (e.g. chamomile, sage, basil), vegetables (e.g. Japanese pepper, pumpkin), buckwheat or sesame, carbohydrates (e.g. honey, brown sugar), salt, miso, flowers. , tea, coffee, cocoa (tea, coffee and cocoa include their preparations), seafood (e.g. oysters, kelp, wakame, bonito flakes), etc.;
Nitrogen sources such as protein decomposition products and yeast extract; other additives such as fragrances, pigments, foaming/foam retention improvers, water quality regulators, and fermentation aids can be used as brewing raw materials. The beer-taste fermented alcoholic beverage of the present invention can use at least malt and hops as raw materials other than brewing water, and in some cases can further use sugars, rice, corn, starch, etc. as raw materials.

In the present invention, a fraction containing peptides with a molecular weight of 2600 to 4600 Da is prepared from grains (for example, malt and/or ungerminated barley or soybean, preferably barley malt or wheat malt), and the fraction is By adding it to a beer-taste fermented alcoholic beverage, the ratio of peptides with a molecular weight of 2,600 to 4,600 Da can be adjusted within a predetermined range,
As a result, it is possible to produce a beer-taste fermented alcoholic beverage with enhanced flavor while suppressing aftertaste. That is, according to the present invention, grains with a molecular weight of 2,600 to 4,600 D
Provided is a method for producing a beer-taste fermented alcoholic beverage, which comprises preparing a fraction containing the peptide of a (gel filtration method for HPLC analysis) and adding the fraction to the beer-taste fermented alcoholic beverage. . The above-mentioned peptide fraction can be added to the beer-taste fermented alcoholic beverage during the manufacturing process of the beverage (for example, the blending process into the fermentation liquid). Peptides with a molecular weight of 2,600 to 4,600 Da (gel filtration method for HPLC analysis) derived from grains can be prepared, for example, by hydrolyzing grains with a proteolytic enzyme such as endo-type protease. It may also be obtained from grain protein hydrolysates, such as hydrolysates.

In the present invention, a beer-flavored fermented alcoholic beverage is produced using malt and/or ungerminated wheat or 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 the fraction to a fermented beer-taste 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 fermented beer-taste alcoholic beverage with enhanced flavor and reduced aftertaste. That is, according to the present invention, there is provided a method for producing a fermented beer-taste alcoholic beverage comprising producing a fermented beer-taste alcoholic beverage using malt and/or ungerminated wheat or barley as at least a part of the raw material, preparing a fraction containing peptides with a molecular weight of 2600 to 4600 Da (gel filtration method for HPLC analysis) from the beverage, and adding the fraction to the fermented beer-taste 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 blending process into the fermentation broth.

Further, according to the present invention, a beer-taste fermented alcoholic beverage with a molecular weight of 2,600 to 460 derived from grains, malt, and/or ungerminated barley as at least a part of the raw materials
A beer-taste fermented alcoholic beverage containing one or more 0 Da peptides is provided, and the beverage is part of the beer-taste fermented alcoholic beverage of the present invention. A beverage containing the peptide has improved or improved flavor as a beer-taste fermented alcoholic beverage, and specifically, is a beverage with enhanced flavor while suppressing aftertaste.

As described above, the beer-taste fermented alcoholic beverage of the present invention can be provided in an embodiment in which the carbohydrate content has been reduced. The carbohydrate content can be reduced by a known method, for example, a method in which glucoamylase is added during the saccharification process, a method in which glucoamylase is added during the fermentation process (
Enzyme Technology Comprehensive Series: From Basics and Analysis to Modification, Advanced Functionalization, and Industrial Use (Edited by Makoto Komiyama,
(see, for example, NTS Corporation, 2010), and a method for reducing carbohydrates by increasing the assimilation rate of yeast using liquid sugar that contains a large amount of carbohydrates that can be assimilated by yeast (see JP 2009-
131202), and a method of filtering wort during the saccharification process (see JP 2012-1477
By using this method (see 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 as the case may be, 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. In addition, when the pH of the beverage is adjusted to less than 4, it is also possible to directly perform the filling step without going through the sterilization step to provide a bottled beverage.

The containers used for the beverage according to the invention may be any containers commonly used for filling beverages, such as metal cans, barrels, plastic bottles (e.g. PET bottles, cups),
Examples include paper containers, bottles, pouch containers, etc., but preferred are metal cans/barrel containers, plastic bottles (for example, PET bottles), and bottles.

According to another aspect of the present invention, there is provided a method for producing a beer-taste fermented alcoholic beverage having improved flavor and using malt and/or ungerminated barley as at least a part of the raw materials, the method comprising:
Adjust the ratio (percentage) of the mass of peptides with a molecular weight of 2,600 to 4,600 Da (gel filtration method for HPLC analysis) in the beverage to 11 to 17% with respect to the total peptide mass of the beverage fractionated by gel filtration method for LC analysis. A method is provided comprising: In the present invention, "improved flavor" or "improved flavor" of a beer-taste fermented alcoholic beverage means to increase the depth of taste while suppressing the aftertaste in the beer-taste fermented alcoholic beverage. The above-mentioned production method of the present invention is a method for producing a beer-taste fermented alcoholic beverage that preferably suppresses aftertaste and enhances the depth of taste. The above-mentioned production 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 its production method.

According to another aspect of the present invention, there is provided a beer-flavored fermented alcoholic beverage having grains or malt and/or ungerminated wheat and barley as at least a part of the raw material, which comprises a fermented alcoholic beverage having a molecular weight of 2,600 to 46
The flavor improver of the present invention is a flavor improver for a fermented beer-taste alcoholic beverage, which comprises a peptide of 0.00 Da (gel filtration method for HPLC analysis) as an active ingredient. The flavor improver for a fermented beer-taste alcoholic beverage is preferably an agent for enhancing the depth of flavor or the drinking response of a fermented beer-taste alcoholic beverage. The flavor improver of the present invention can be produced according to the description of the fermented beer-taste 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 the raw materials. The flavor improvement method of the present invention provides a ratio (percentage) of the mass of peptides with a molecular weight of 2600 to 4600 Da (gel filtration method for HPLC analysis) in the beverage to the total peptide mass of the beverage fractionated by gel filtration method for HPLC analysis. ) by adjusting it to 11 to 17%. The flavor improving method of the present invention is preferably a method of enhancing the depth of taste of a beer-taste fermented alcoholic beverage or a method of enhancing the drinking sensation of the beverage. The flavor improvement method of the present invention can be carried out according to the description regarding the beer-taste fermented alcoholic beverage of the present invention and its manufacturing method.

The present invention will be explained 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
(1) Production of beer-flavored fermented alcoholic beverages (Beer Base 1 and 2) 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.
Parts by weight of the enzyme preparation were added and held for 40 minutes, then the temperature was raised to 78°C and held for 5 minutes, and then filtered to obtain wort.

Following the above wort preparation step, 19.0 parts by mass of hops and liquid sugar were added to the obtained wort and boiled at 100°C for 90 minutes, and the wort was allowed to stand, the trub was separated, and then cooled to obtain a pre-fermentation liquid. Then, bottom-fermenting yeast was added to the pre-fermentation liquid, and main fermentation and post-fermentation were carried out according to a conventional method. Next, the fermentation liquid after the post-fermentation was stored by maintaining it at a lower temperature, and filtered to obtain a clear beer-flavored alcoholic beverage (malt usage ratio 50%) (beer base 1) with a sugar 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, 10.0 parts by mass of barley malt was mixed with 100 parts by mass of hot water at 64°C.
The mixture was heated to 78° C. and held for 5 minutes, and then filtered to obtain wort.

Following the above wort preparation process, 9.5 parts by mass of liquid sugar mainly composed of hops and assimilable sugars was added to the obtained wort, and after boiling at 100°C for 90 minutes, the wort was allowed to stand still. After separating the trives, the mixture was cooled to obtain a pre-fermentation liquid. Thereafter, bottom-fermenting yeast was added to the pre-fermentation solution, and main fermentation and post-fermentation were performed according to a conventional method. Subsequently, the fermented liquid after post-fermentation is stored by maintaining it at a lower temperature and filtered to produce a clear beer-taste alcoholic beverage (50% malt usage ratio) with a carbohydrate concentration of 0.3 g/100 mL ( Beer base 2) was obtained.

(2) Preparation of peptide fractions A commercially available beer-flavored beverage was degassed, weighed, and freeze-dried.
The resulting solution was concentrated 5-fold in 100 mM NaCl solution to prepare a sample for gel filtration fractionation. 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: 100 mM NaCl
Flow rate: 2.5 mL/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 fraction (fraction number: F10-11) was purified under the following conditions and quantified.

The obtained fractions were extracted using a solid phase extraction column (Bond Elute C18, Agilent technolo
Gies, Inc.), washed with demineralized water, eluted with a 50% ethanol aqueous solution, concentrated to dryness, and condensed with demineralized water to obtain a concentrated solution. This was designated as a peptide fraction.

Protein quantification of the peptide fraction obtained by purification of the peptide fraction can be performed using a commercially available kit (
It was carried out by the Lowry method using a DC protein assay (manufactured by Bio-Rad). first,
The concentration of the fractionated solution was adjusted to an appropriate range. To 5 μL of the concentration-adjusted sample, 50 μL of Solution A was added and stirred, and then 400 μL of Solution B was added and stirred. After carrying out a color reaction for 15 minutes at room temperature, 350 μL of the mixture 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 calibration curve prepared in advance. Note that the calibration curve was created using BSA (bovine serum albumin).

(3) Preparation of gel filtration fraction for HPLC analysis Regarding the sample brew product, the sample sample obtained in (1) above was degassed, weighed, and freeze-dried. Each lyophilizate was dissolved in 50mM sodium phosphate buffer (pH 7.0, 150mM N
(contains aCl) to prepare a 2.5-fold concentrated solution, which was used as a sample for gel filtration fractionation. The peptide fraction obtained in (2) above was dissolved in the above buffer to a concentration equivalent to a 10-fold concentrate, and used as a sample for gel filtration fractionation. Each of the fractionated samples was subjected to gel filtration fractionation under the following conditions to obtain fractionated fractions (fractions 1 to 10).

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

Fractionation program:

Protein quantification of the fractionated solution was performed by the Lowry method described in (2) above.

(4) Measurement of molecular weight by gel filtration method for HPLC analysis Sample preparation for gel filtration for HPLC analysis The fractions obtained in (3) above (fractions 1 to 10) were used as samples.

B Creation of a molecular weight calibration curve Inject 50 μL of a peptide with a known molecular weight dissolved in ultrapure water at 0.1 to 5 mg/mL as appropriate under the column, eluent, flow rate, and detection wavelength described in the gel filtration conditions for HPLC analysis above. HPLC analysis was performed to confirm the retention time (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 a molecular weight of 2,600 to 4,600 Da The ratio of peptides with a molecular weight of 2,600 to 4,600 Da was calculated using the following calculation formula using the peptide concentration corresponding to the product in each fraction.

(6) Analysis of Carbohydrate Concentration and Original Extract Concentration The carbohydrate concentration of the sample was measured according to the method described in the 5th Edition of the Analysis Manual for the Standard Tables of Food Composition in Japan, based on the Nutrition Labeling Standards.
Reduced pressure 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 according to the BCOJ Beer Analysis Method (Beer Brewing 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 produced in (1) above (Beer Bases 1 and 2)
A test group was set up 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 carried out by four trained panelists. Specifically, the "depth of flavor" and "aftertaste" were evaluated on a nine-point scale from 1 to 9, with one-point increments, and the average evaluation 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. In addition, "aftertaste" refers to the flavor sensation consisting of the five basic tastes and stimulating tastes (sweetness, sourness, saltiness, umami, bitterness, astringency, spiciness, roughness, etc.) that remain in the mouth after swallowing.

Test example 1 evaluating the effect of adding peptide fraction to beer base 1 (test groups 1 to 5)
In this case, evaluation was performed with the scores of "thickness of taste" and "aftertaste" of test group 1 fixed at 3 in advance. In addition, in Test Example 2 (Test Groups 6 to 10) in which the effect of adding the peptide fraction to Beer Base 2 was evaluated, the scores for "Taste Thickness" and "Aftertaste" of Test Group 6 were fixed at 3 in advance. I did it. In all test examples, the thickness of the taste was 3.5 points or higher, and the aftertaste was 5.0 points.
The following was determined to be a preferable embodiment.

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

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

In addition, when we conducted the above sensory evaluation test for Beer Bases 1 and 2 (however, the scores for "thickness of taste" and "aftertaste" were not fixed at 3 in advance), we obtained the following results. Ta.

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 lose rich flavor, adjusting the ratio of 2600-4600 Da peptides within a specified range can increase the richness of flavor while suppressing the aftertaste.

Claims (1)

A beer-flavored fermented alcoholic beverage that uses malt as at least a part of the raw material, wherein 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 is 11 to 17%, the concentration of the peptides having a molecular weight of 2600 to 4600 Da (gel filtration for HPLC analysis) in the beverage 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 that uses malt as at least a part of the raw material.

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