JP7392011B2 - Beer-flavored fermented alcoholic beverage with zero carbohydrates - Google Patents

Description

The present invention relates to a beer-taste fermented alcoholic beverage with zero carbohydrates, and more particularly to a beer-taste fermented alcoholic beverage with a carbohydrate concentration of less than 0.5 g/100mL.

As health consciousness has increased in recent years, there has been a demand for beer-flavored beverages with reduced sugar content. To date, various technologies have been developed with the aim of producing beer-flavored beverages with reduced sugar content.

For example, as a technique for stably producing a low-carbohydrate beer-taste fermented alcoholic beverage, a technique using assimilable monosaccharides and assimilable disaccharides as sugar sources is known (Patent Document 1). Furthermore, as a technique for producing a low-carbohydrate beer-taste beverage with a significantly low risk of microbial contamination, a technique is known in which the oligosaccharide content in carbohydrate raw materials is used within a specific range (Patent Document 2).

Japanese Patent Application Publication No. 2009-131202 Japanese Patent Application Publication No. 2012-157323

Beer-taste fermented alcoholic beverage with reduced sugar content, sugar concentration 0.5mg/1
So-called "sugar-free" beer-taste fermented alcoholic beverages with a volume of less than 0.0 mL have a considerable amount of carbohydrates reduced, and therefore have a particularly watery and weak flavor impression. Therefore, if a rich flavor can be achieved in a beer-taste fermented alcoholic beverage with zero carbohydrates, it is possible to provide a beer-taste fermented alcoholic beverage that is satisfying to drink despite having zero carbohydrates.

That is, an object of the present invention is to provide a beer-taste fermented alcoholic beverage with zero carbohydrates and a rich taste.

The present inventors have discovered that in a beer-taste fermented alcoholic beverage whose carbohydrate concentration has been reduced to less than 0.5 mg/100 mL, the ratio of peptides having a molecular weight of 800 to 1500 Da is within a specific range. We discovered that it is possible to achieve a richer taste. The present invention is based on this knowledge.

According to the present invention, the following inventions are provided.
[1] At least a part of the raw material is malt and/or ungerminated barley, and the carbohydrate concentration is 0.
A beer-taste fermented alcoholic beverage of less than 5 g/100 mL, with a molecular weight of 800 to 800, relative to the mass of all peptides in the beverage fractionated by gel filtration for HPLC analysis.
The mass ratio (percentage) of a peptide of 1500 Da (gel filtration method for HPLC analysis) is 17~
45% beer-taste fermented alcoholic beverage.
[2] The beer-taste fermented alcoholic beverage according to [1] above, wherein the mass ratio of the peptide is 18 to 25%.
[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 with improved flavor and a carbohydrate concentration of less than 0.5 g/100 mL, using malt and/or ungerminated barley as at least a part of the raw materials, The ratio (percentage) of the mass of peptides with a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the beverage to the mass of all peptides in the beverage fractionated by gel filtration method for HPLC analysis is 17 to 45%. A method including adjusting.
[5] In the manufacturing process of beer-taste fermented alcoholic beverages, molecular weights of 800 to 1500 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] Molecular weight 800 to 1500 Da (gel filtration method for HPLC analysis) contained in beer-taste fermented alcoholic beverages containing malt and/or ungerminated barley as at least a part of the raw materials
A flavor improving agent for a beer-taste fermented alcoholic beverage with a carbohydrate concentration of less than 0.5 g/100 mL, which comprises a peptide as an active ingredient.
[7] At least a part of the raw material is malt and/or ungerminated barley, and the carbohydrate concentration is 0.
A method for improving the flavor of a beer-taste fermented alcoholic beverage of less than 5 g/100 mL, the method comprising:
The ratio (percentage) of the mass of peptides with a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the beverage to the mass of all peptides in the beverage fractionated by gel filtration method for PLC analysis is 17 to 45%. A method including adjusting.

According to the present invention, in a beer-taste fermented alcoholic beverage with a carbohydrate concentration of less than 0.5 g/100 mL, a rich taste is achieved by adjusting the ratio of peptides with a molecular weight of 800 to 1,500 Da within a specific range. It is advantageous in that it can provide beer-taste fermented alcoholic beverages with zero quality and can meet the diverse needs of consumers, such as health-consciousness.

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. FIG. 2 is a diagram showing the sensory evaluation results (average value ± standard error of n=5) of Example 1. The influence of the addition of a peptide fraction with a molecular weight of 800 to 1500 Da on the depth of taste and astringency was shown using sensory evaluation scores.

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 "beer-taste fermented alcoholic beverage" is a beer-taste fermented alcoholic beverage fermented by yeast using a carbon source, a nitrogen source, water, etc. as raw materials,
Beer, low-malt beer, and beverages made by adding alcohol to beer or low-malt beer that uses malt as an ingredient (e.g., a new genre of liqueur beverages classified as "liqueur (sparkling) (1)" under the Liquor Tax Act) can be mentioned.

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 beer-taste fermented alcoholic beverage of the present invention is a carbohydrate-free beverage with a reduced carbohydrate concentration. Specifically, the beer-taste fermented alcoholic beverage of the present invention has a carbohydrate concentration of 0.5
g/100mL.

Carbohydrate concentration can be measured by a known method, and is calculated by excluding water, protein, fat, ash, and dietary fiber from the mass of the sample (according to the Nutrition Labeling Standards (2009)).
This can be done in accordance with the Consumer Affairs Agency Notification No. 9 (partially revised) dated December 16).

In the beer-taste fermented alcoholic beverage of the present invention, the molecular weight in the beverage is 800 to 1500 Da (HP
Gel filtration method for LC analysis) is characterized in that the mass ratio (percentage) of the peptide is within a specific range. The ratio is the concentration of peptides (mg/mL) with a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the beverage to the concentration of total peptides derived from the beverage (mg/mL) fractionated by gel filtration method for HPLC analysis. It can be calculated by dividing by In the present invention,
The lower limit of the peptide ratio (more than or above) is 17% (from the perspective of imparting depth of taste).
The upper limit (less than or equal to) can be 45% (preferably 25%). These lower and upper limits can be combined arbitrarily, for example, the range of the peptide ratio is 17 to 45%, preferably 18 to 25%, more preferably 18% to less than 25%. can do.

In the beer-taste fermented alcoholic beverage of the present invention, the molecular weight in the beverage is 800 to 150.
The lower limit of the peptide concentration of 0 Da (gel filtration method for HPLC analysis) is set to 0
．． 10 mg/mL (preferably 0.15 mg/mL), and the upper limit (
less than or equal to 2.50 mg/mL (preferably 2.00 mg/mL, 1.50 mg)
/mL, 1.20mg/mL, 1.00mg/mL, 0.70mg/mL or 0.65
mg/mL). These lower and upper limits can be arbitrarily combined. For example, the concentration of peptides with a molecular weight of 800 to 1,500 Da (gel filtration method for HPLC analysis) in beverages is 0.10 to 2.50 mg/mL or 0.15 mg/mL. ～1.20mg
/mL.

In the beer-taste fermented alcoholic beverage of the present invention, the concentration of peptides with a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the beverage (mg/mL) relative to the original extract concentration (OE concentration) (°P) in the beverage The lower limit of the ratio (greater than or greater than) of 0.
0.02 (preferably 0.03), and the upper limit (less than or equal to) 0.02 (preferably 0.03).
50 (preferably 0.30, 0.15, 0.07 or 0.05). These lower and upper limits can be arbitrarily combined, for example, the concentration of peptides with a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the beverage (mg/mL) relative to the OE concentration (°P) in the beverage. ) is set at a ratio of 0.02 to 0.50 (preferably 0.02 to 0.50).
03 to 0.30).

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 a rich taste despite having a reduced carbohydrate concentration. Here, "thickness of taste" refers to the flavor sensation perceived by the breadth of taste, complexity, imparting a sense of body, etc.

The beer-taste fermented alcoholic beverage of the present invention can be used as a normal beer as long as the carbohydrate concentration in the beverage is reduced within a predetermined range and the peptide ratio of 800 to 1500 Da in the beverage is adjusted within a predetermined range. It can be manufactured according to the manufacturing procedure for flavored fermented alcoholic beverages.

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. I can do it. 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 80.
By increasing the ratio of peptides ranging from 0 to 1500 Da (gel filtration method for HPLC analysis), it is possible to adjust the ratio to within a predetermined value range, resulting in beer-taste fermentation that has no carbohydrates yet has a richer taste. Alcoholic beverages can be produced. 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 can be used as part of the raw material. In the beer-taste fermented alcoholic beverage of the present invention, ungerminated barley (including extracts) and ungerminated wheat (including extracts) may be used as raw materials. I can do it.

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 800 to 1,500 Da is prepared from grains (for example, malt and/or ungerminated barley and soybeans), and the fraction is converted into a sugar-free beer-taste beverage. By adding it, the ratio of peptides with a molecular weight of 800 to 1500 Da can be adjusted within a predetermined value range, and as a result, a beer-taste fermented alcoholic beverage with a rich taste can be produced while being free of carbohydrates. I can do it. That is, according to the present invention, a fraction containing peptides with a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) is prepared from grains, and the fraction is added to beer with a carbohydrate concentration of less than 0.5 g/100 mL. A method for producing a beer-taste fermented alcoholic beverage is provided, the method comprising adding the beer-taste fermented alcoholic beverage 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 800 to 1500 Da (gel filtration method for HPLC analysis) derived from grains can be prepared, for example, by hydrolyzing grains with a proteolytic enzyme such as peptidase, and by hydrolyzing soybean protein. It may also be obtained from protein hydrolysates of grains such as grains.

In the present invention, it is also possible to produce a beer-taste fermented alcoholic beverage using malt and/or ungerminated barley as at least a part of the raw materials,
By preparing a fraction containing peptides and adding this fraction to a sugar-free beer-taste beverage, the ratio of peptides with a molecular weight of 800 to 1500 Da can be adjusted within a predetermined range, and as a result, It is possible to produce a beer-taste fermented alcoholic beverage with no sugar content and a rich taste. That is, according to the present invention, a beer-taste fermented alcoholic beverage containing malt and/or ungerminated barley as at least a part of the raw materials is produced, and then the beverage is processed using gel filtration method for HPLC analysis (with a molecular weight of 800 to 1500 Da). )
Provided is a method for producing a beer-taste fermented alcoholic beverage, which comprises preparing a fraction containing a peptide and adding the fraction to a beer-taste fermented alcoholic beverage having a carbohydrate concentration of less than 0.5 g/100 mL. be done. 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).

Further, according to the present invention, a beer-taste fermented alcoholic beverage with a molecular weight of 800 to 1500 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 peptides of Da 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 enhanced flavor as a beer-taste fermented alcoholic beverage, and specifically, is a beverage with enhanced flavor.

Reduction of carbohydrates in beer-taste fermented alcoholic beverages can be carried out according to known methods, for example, a method of adding glucoamylase during the saccharification process, a method of adding glucoamylase during the fermentation process (Enzyme Utilization Technology System Basics)・From analysis to modification, advanced functionality, and industrial use (see Makoto Komiyama's supervision, NTS Co., Ltd., 2010), etc.) Sugar in beverages can be reduced by increasing assimilation (see Japanese Patent Application Laid-open No. 2009-131202), and by filtering wort during the saccharification process (see Japanese Patent Application Laid-Open No. 2012-147780). The quality concentration can be set to a predetermined value.

The beverage provided by the present invention may be subjected to a step of adding carbon dioxide gas, as the case may be, and may be provided as a packaged beverage through steps such as a filling step and a sterilization step. Sterilization may be performed before or after filling the container. Furthermore, if the pH of the beverage is adjusted to less than 4, the beverage can be packed into containers by directly performing the filling process without going through the sterilization process.

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 with zero carbohydrates, which uses malt and/or ungerminated barley as at least a part of the raw materials and has improved flavor, The ratio (percentage) of the mass of peptides with a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the beverage to the mass of all peptides in the beverage fractionated by gel filtration method for HPLC analysis is 17 to 45%. A method is provided comprising adjusting. "Improved flavor" or "improved flavor" of beer-taste fermented alcoholic beverage according to the present invention
This means that a rich taste is achieved in a beer-taste fermented alcoholic beverage with zero carbohydrates. The above-mentioned production method of the present invention is preferably a method for producing a beer-like fermented alcoholic beverage with zero carbohydrates and imparted with a rich 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, the beer-taste fermented alcoholic beverage containing grains or malt and/or ungerminated barley as at least a part of the raw materials has a molecular weight of 800 to 150.
Provided is a flavor improving agent for beer-taste fermented alcoholic beverages with zero carbohydrates, which contains a 0 Da (gel filtration method for HPLC analysis) peptide as an active ingredient. The flavor improving agent for beer-taste fermented alcoholic beverages is preferably an agent that thickens the taste of beer-taste fermented alcoholic beverages with zero carbohydrates. The flavor improving agent of the present invention can be produced according to the description regarding the beer-taste fermented alcoholic beverage of the present invention and its manufacturing method.

According to yet another aspect of the present invention, there is provided a method for improving the flavor of a sugar-free 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 is characterized in that the ratio of the mass of peptides with a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the beverage to the mass of all peptides in the beverage fractionated by gel filtration method for HPLC analysis ( This can be carried out by adjusting the percentage) to 17 to 45%. The flavor improvement method of the present invention is a method for imparting a richer taste to a beer-taste fermented alcoholic beverage, preferably with zero carbohydrates. 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.

Analysis of Carbohydrate Concentration Carbohydrate concentration was measured based on the nutrition labeling standards in accordance with the method described in the well-known fifth edition Japanese Food Standard Composition Table Analysis Manual. That is, from the mass of the sample drink, water (vacuum heat drying method), protein (method using an automatic analyzer), lipid (Soxhlet extraction method (3)),
It was measured by a calculation method excluding ash content (muffle furnace combustion method) and dietary fiber content (Prosky enzyme gravimetric method).

Reference Example 1: Identification of a peptide fraction that imparts a desirable flavor to a beer-taste beverage (1) Production of a beer-taste fermented alcoholic beverage A beer-taste fermented alcoholic beverage is produced by an infusion method using barley malt, hops, and an enzyme preparation. Manufactured. Specifically, 100 g of barley malt was added to 300 mL of 65° C. hot water, held for 60 minutes, further heated to 78° C., held for 5 minutes, and then filtered to obtain wort. Subsequently, 0.8 g/L of hops was added and boiled at 100°C for 90 minutes, followed by filtration to obtain a pre-fermentation liquid.

Thereafter, fermentation was performed using beer yeast according to a conventional method, and the flavor of the fermented liquid was confirmed. As a result, it was confirmed that the resulting fermented liquid had suppressed unpleasant tastes, had a soft and smooth texture similar to beer, and had a harmonious taste.

(2) Gel filtration fractionation The fermentation liquid obtained in the above (1) was filtered with a 0.45 μm filter, and the filtered fermentation liquid was weighed and freeze-dried. The dried product was dissolved in a 100 mM NaCl solution to prepare a 5-fold concentrated solution, which was used as a sample for fractionation. The sample was subjected to gel filtration fractionation under the following conditions.

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

Based on the sensory evaluation of the fractions, the fractions were divided into nine groups, pre-fraction, A1, A2, B, C, D, E, F, and G, as shown in Table 1, according to differences in flavor characteristics. In addition, the flavor characteristics of each fraction were determined through discussion among five trained panelists.

(3) Purification of peptide fraction Fractions C and D obtained in (2) above were purified using a solid phase extraction column (Bond Elute C).
18, manufactured by Agilent Technologies), washed with demineralized water, eluted with 50% ethanol aqueous solution, concentrated to dryness, condensed with demineralized water, and concentrated. did. This was designated as a peptide fraction.

(4) Protein quantification by the Lowry method The protein quantification of the peptide fraction obtained by the gel filtration fraction in (2) above and the peptide fraction purified in (3) above can be carried out using commercially available kits (DC protein assay, Bio- The Lowry method was used (manufactured by 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 15 minutes of color reaction at room temperature, 3 cells were added to a 96-well plate.
50 μL was transferred 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).

(5) Results The peptide fractions were analyzed using a Superdex 75 10/300 column and the molecular weights were estimated, and the peptides in fractions C and D were found to have a molecular weight distribution of about 800 to 1500 Da.

From the above results, the molecular weight of the fractionated and purified beer-taste fermented alcoholic beverage is approximately 80.
It was suggested that peptide fractions C and D ranging from 0 to 1500 Da are effective in imparting drinking sensation (ie, depth of taste).

Example 1: Production of sugar-free beer-taste fermented alcoholic beverage and addition of peptides to the beverage
Analysis of the influence of addition on flavor depth (1) Production of beer-taste fermented alcoholic beverage with zero carbohydrates In the production of the beer-taste fermented alcoholic beverage of this example, barley malt was used as the main raw material. During saccharification, an enzyme preparation was used, the temperature and time of saccharification was adjusted, and wort was obtained by filtration. Specifically, 10 parts by mass of barley malt, a β-glucanase enzyme preparation, and an enzyme preparation mainly consisting of glucoamylase were added to 100 parts by mass of hot water at 50°C.
After holding for 0 minutes, the temperature was raised to 64°C and held for 70 minutes. Thereafter, the temperature was raised to 78°C, 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 was stored by maintaining it at a lower temperature and filtered to obtain a clear beer-taste alcoholic beverage.

(2) Preparation of peptide fraction A commercially available beer-taste beverage was degassed, weighed, and freeze-dried. 1 lyophilized product
A 5-fold concentrated solution was prepared by dissolving in 00 mM NaCl solution and used as a sample for gel filtration fractionation. Gel filtration fractionation was carried out as described in Reference Example 1 (2), and the obtained fraction C (fraction number: F25-29) was purified as described in Reference Example 1 (3). At this time, in order to improve the recovery rate in purification, the fraction that passed through the solid phase extraction column was also collected using a synthetic adsorbent (Sepabead SP207, manufactured by Mitsubishi Chemical Corporation). Protein quantification was performed by the Lowry method described in Reference Example 1 (4).

(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 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), 20% (v/v) acetonitrile, 150mM NaCl
Flow rate: 0.5mL/min (constant flow rate)
Detection wavelength: 215nm

Protein quantification of the fractionated solution was performed by the Lowry method described in Reference Example 1 (4).

(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 7 of fractions 1 to 10 had a molecular weight in the range of 800 to 1500 Da.

(5) Calculation of the ratio of peptides with a molecular weight of 800 to 1,500 Da The ratio of peptides with a molecular weight of 800 to 1,500 Da was calculated using the following calculation formula using the peptide concentration corresponding to the product in each fraction.

The sample peptide ratios were as shown in Table 3.

(6) Analysis of carbohydrate concentration and original extract concentration The carbohydrate concentration of the sample brewed product was measured based on the nutritional labeling standards as described above. The results were as shown in Table 3. In addition, the original extract concentration (OE concentration) of the sample brewed product was measured according to the BCOJ beer analysis method (Beer Brewing Association (2013), 8.3.6, 8.4.3, and 8.5. This was carried out using an alcoholizer (manufactured by Anton Paar).

(7) Sensory evaluation Test in which the peptide fraction obtained in (2) above was not added to the beer-taste fermented alcoholic beverage (malt usage ratio 50%) produced in (1) above and in a test group (trial brew product). Sample drinks (sample numbers 1 to 10) with peptide concentrations of 800 to 1500 Da adjusted as shown in Table 3 were prepared.

Sensory evaluation was conducted by five trained panelists. Specifically, "thickness of taste" and "offensive taste" were evaluated on a 17-point scale from 1 to 9 in 0.5-point increments, and the average value of the evaluation scores of the five panelists was calculated. Here, "thickness of taste" refers to the flavor sensation perceived by the breadth of taste, complexity, imparting body feeling, etc. Sample number 1 for “thickness of taste”
The sample drink of Sample No. 6 was given a score of 1 (no thick taste), and the sample drink of sample number 6 was given a score of 5 (good taste), and a score of 1.5 or more was judged as having an improved taste. In addition, "miscellaneous taste" refers to astringent, harsh, and astringent tastes. For "miscellaneous taste", add 1 sample drink of sample number 1.
The sample drink of sample number 6 was given a score of 3 (some astringency), and a score of 4.0 or more was judged as improved astringency.

Based on the sensory evaluation results regarding "thickness of flavor" and "presence or absence of off-taste", comprehensive evaluation was performed using the following criteria.
◎: Improved flavor depth (2.5 points or more) and less unpleasant taste (less than 4.0 points)
○: Thickness of taste is slightly improved (1.5 points or more) and there is little off-taste (less than 4.0 points), or the depth of taste is improved (over 2.5 points) but there is some off-taste. (4.0 points or more)
×: Thickness of taste has not improved (less than 1.5 points)

The results of the sensory evaluation were as shown in Tables 3 and 4 and FIG. 2.

From the results in Table 3, even in a sugar-free beer-taste fermented alcoholic beverage with a sugar concentration of less than 0.5g/100mL (50% malt usage ratio), increasing the ratio of 800 to 1500 Da peptides will improve the taste. It was confirmed that the "thickness" was improved.

Claims (9)

A beer-taste fermented alcoholic beverage containing barley malt as at least a part of the raw material and having a carbohydrate concentration of less than 0.5 g/100 mL, based on the mass of total peptides of the beverage fractionated by gel filtration method for HPLC analysis. A beer in which the mass ratio (percentage) of peptides with a molecular weight of 800 to 1,500 Da (gel filtration method for HPLC analysis) in the beverage is 17 to 45%, and the peptide is made from barley malt as at least a part of the raw material. A beer-taste fermented alcoholic beverage derived from a fermented-taste alcoholic beverage. The beer-taste fermented alcoholic beverage according to claim 1, wherein the mass ratio of the peptide is 18 to 25%. The beer-taste fermented alcoholic beverage according to claim 1 or 2, wherein the malt usage ratio is 50% or more. The original extract concentration of the drink is 4°P or more and 14°P or less, and/or the concentration of peptides with a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the drink is 0.15 mg/mL or more. The beer-taste fermented alcoholic beverage according to any one of claims 1 to 3, which has a concentration of .65 mg/mL or less. A method for producing a beer-taste fermented alcoholic beverage with an improved flavor and a sugar concentration of less than 0.5 g/100 mL, using barley malt as at least a part of the raw material, the method comprising: fractionating the beverage by gel filtration for HPLC analysis; adjusting the ratio (percentage) of the mass of peptides with a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the beverage to 17 to 45% with respect to the mass of all peptides in the beverage, and A manufacturing method, wherein the peptide is derived from a beer-taste fermented alcoholic beverage containing barley malt as at least a part of the raw material. A claim that includes a step of adding a peptide with a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) contained in a beer-taste fermented alcoholic beverage containing barley malt as at least a part of the raw material, in the manufacturing process of a beer-taste fermented alcoholic beverage. The manufacturing method according to item 5. The original extract concentration of the drink is 4°P or more and 10°P or less, and/or the concentration of peptides with a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the drink is 0.15 mg/mL or more. The manufacturing method according to claim 5 or 6, wherein the concentration is .65 mg/mL or less. A method for improving the flavor of a beer-taste fermented alcoholic beverage with a carbohydrate concentration of less than 0.5 g/100 mL, which uses barley malt as at least a part of the raw material, wherein the entire content of the beverage is fractionated by a gel filtration method for HPLC analysis. adjusting the mass ratio (percentage) of a peptide having a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the beverage to the mass of the peptide to 17 to 45%; a beer-taste fermented alcoholic beverage at least in part as an ingredient. The original extract concentration of the drink is 4°P or more and 10°P or less, and/or the concentration of peptides with a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the drink is 0.15 mg/mL or more. 9. The method of claim 8, wherein the amount is .65 mg/mL or less.

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