JP7103980B2 - Beer-taste fermented alcoholic beverage with zero sugar - Google Patents

Description

The present invention relates to a sugar-free beer-taste fermented alcoholic beverage, and more particularly to a beer-taste fermented alcoholic beverage having a sugar concentration of less than 0.5 g / 100 mL.

Due to the growing health consciousness in recent years, beer-taste beverages having a reduced sugar content are required. So far, various techniques have been developed aiming at beer-taste beverages having a reduced sugar content.

For example, as a technique for stably producing a low-carbohydrate beer-taste fermented alcoholic beverage, a technique using assimilating monosaccharides and assimilating disaccharides as sugar sources is known (Patent Document 1). Further, as a technique for producing a low-carbohydrate beer-taste beverage by a method having a remarkably low risk of microbial contamination, a technique of using the oligosaccharide content in a sugar raw material within a specific range is known (Patent Document 2).

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

Among the beer-taste fermented alcoholic beverages with reduced sugar, the so-called "zero sugar" beer-taste fermented alcoholic beverages with a sugar concentration of less than 0.5 mg / 100 mL have a considerable amount of sugar reduced. Especially, it gives a watery and light flavor impression. Therefore, if the thickness of the taste can be realized in a beer-taste fermented alcoholic beverage having zero sugar, it is possible to provide a beer-taste fermented alcoholic beverage having zero sugar and a satisfying drink.

That is, an object of the present invention is to provide a sugar-free beer-taste fermented alcoholic beverage having a thick taste.

The present inventors set the ratio of peptides having a molecular weight of 800 to 1500 Da within a specific range in a beer-taste fermented alcoholic beverage in which the sugar concentration is reduced to less than 0.5 mg / 100 mL. It was found that the thickness of the taste can be realized. 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 at least a part of malt and / or ungerminated wheat as a raw material and having a sugar concentration of less than 0.5 g / 100 mL, which is fractionated by a gel filtration method for HPLC analysis. A beer-taste fermented alcoholic beverage in which the ratio (percentage) of the mass of peptides having 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 is 17 to 45%.
[2] The beer-taste fermented alcoholic beverage according to the above [1], wherein the ratio of the mass of the peptide is 18 to 25%.
[3] The beer-taste fermented alcoholic beverage according to the above [1] or [2], wherein the malt usage ratio is 50% or more.
[4] A method for producing a beer-taste fermented alcoholic beverage having an improved flavor, using malt and / or ungerminated wheat as a raw material and having a sugar concentration of less than 0.5 g / 100 mL. The ratio (percentage) of the mass of the peptide having a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the beverage to the mass of all the peptides in the beverage fractionated by the gel filtration method for HPLC analysis is 17 to 45%. A method that involves adjusting.
[5] In the process of producing a beer-taste fermented alcoholic beverage, a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) contained in a beer-taste fermented alcoholic beverage containing at least a part of malt and / or ungerminated wheat as a raw material. The production method according to the above [4], which comprises a step of adding the peptide of.
[6] A peptide having a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) contained in a beer-taste fermented alcoholic beverage containing at least a part of malt and / or ungerminated wheat as a raw material is contained as an active ingredient. A flavor improver for beer-taste fermented alcoholic beverages with a sugar concentration of less than 0.5 g / 100 mL.
[7] A method for improving the flavor of a beer-taste fermented alcoholic beverage having a sugar concentration of less than 0.5 g / 100 mL, which uses malt and / or ungerminated wheat as at least a part of the raw material, and is a gel filtration for HPLC analysis. Including adjusting the ratio (percentage) of the mass of the peptide having a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the beverage to the mass of the total peptide of the beverage fractionated by the method to 17 to 45%. How to be.

According to the present invention, in a beer-taste fermented alcoholic beverage having a sugar concentration of less than 0.5 g / 100 mL, a sugar whose taste thickness is realized by adjusting the ratio of peptides having a molecular weight of 800 to 1500 Da within a specific range. It is advantageous in that it can provide fermented alcoholic beverages with a beer taste of zero quality and can meet various needs such as consumer health consciousness.

It is a figure which showed the calibration curve prepared from the holding time of the gel filtration method for HPLC analysis and the molecular weight of a known substance carried out in Example 1. FIG. It is a figure which showed the sensory evaluation result (mean value ± standard error of n = 5) of Example 1. FIG. The effect of the addition of the peptide fraction having a molecular weight of 800 to 1500 Da on the thickness and astringency of the taste was shown by the sensory evaluation score.

Specific description of the invention

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

In the present invention, the "beer-taste fermented alcoholic beverage" is a beer-taste fermented alcoholic beverage fermented with yeast using carbon sources, nitrogen sources, water and the like as raw materials, and uses malt as beer, sparkling liquor and raw materials. Beverages obtained by adding alcohol to beer or sparkling liquor (for example, liqueur-based new genre beverages classified as "liqueur (foaming) (1)" under the Liquor Tax Law) can be mentioned.

The beer-taste fermented alcoholic beverage of the present invention uses malt and / or ungerminated wheat as a part of the raw material, and any malt usage ratio can be taken. For example, the malt usage ratio is 50. It can be less than%, 50% or more, less than 67%, 67% or more, or 95% or more. In the present specification, the "malt use ratio" means the ratio of the malt mass to the mass of all raw materials excluding brewing water.

The beer-taste fermented alcoholic beverage of the present invention is a sugar-free beverage with a reduced sugar concentration. Specifically, the sugar concentration of the beer-taste fermented alcoholic beverage of the present invention is less than 0.5 g / 100 mL. Is.

The carbohydrate concentration can be measured by a known method, and is calculated by excluding water, protein, lipid, ash and dietary fiber from the mass of the sample (Nutrition Labeling Standard (December 16, 2009). It can be done in accordance with (Refer to Notification No. 9 Partial Amendment) of the Consumer Affairs Agency).

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

In the beer-taste fermented alcoholic beverage of the present invention, the lower limit (or more or more) of the peptide concentration having a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the beverage is 0.10 mg / mL (preferably 0.15 mg /). mL), and the upper limit (or less or less) is 2.50 mg / mL (preferably 2.00 mg / mL, 1.50 mg / mL, 1.20 mg / mL, 1.00 mg / mL, It can be 0.70 mg / mL or 0.65 mg / mL). These lower and upper limits can be arbitrarily combined, and for example, the peptide concentration of a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in a beverage is 0.10 to 2.50 mg / mL or 0.15. It can be ~ 1.20 mg / mL.

The beer-taste fermented alcoholic beverage of the present invention also has a peptide concentration (mg / mL) having a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the beverage with respect to the original extract concentration (OE concentration) (° P) in the beverage. The lower limit (preferably 0.03) of the ratio of the above can be 0.02 (preferably 0.03), and the upper limit (less than or less than) of 0.50 (preferably 0.30, 0.). 15, 0.07 or 0.05). These lower and upper limits can be arbitrarily combined, and for example, the peptide concentration (mg / mL) of the molecular weight in the beverage of 800 to 1500 Da (gel filtration method for HPLC analysis) with respect to the OE concentration (° P) in the beverage. ) Can be 0.02 to 0.50 (preferably 0.03 to 0.30).

The lower limit (or more or more) 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 (less than or less than) is 20 ° P. (Preferably 14 ° P, more preferably 10 ° P). These lower and upper limits can be arbitrarily combined, and for example, the OE concentration in the beer-taste fermented alcoholic beverage of the present invention can be 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 alcoholizer (manufactured by Anton Pearl Co., Ltd.)). In a beer-taste fermented alcoholic beverage using malt and / or ungerminated wheat as a raw material, the "original extract concentration" can be rephrased as the "raw wort extract concentration".

The molecular weight of peptides in beverages is measured by gel filtration by high performance liquid chromatography (sometimes referred to herein as "gel filtration for HPLC analysis"). Specifically, the molecular weight can be calculated from the retention time using the calibration curve of the gel filtration method for HPLC analysis (for example, FIG. 1), and samples having different molecular weights can be sorted according to the retention time. Specific examples of the measurement of the molecular weight by the gel filtration method for HPLC analysis are as shown in Example 1 below. Further, the quantification of the peptide can be carried out by the Lowry method (Lowry method).

The beer-taste fermented alcoholic beverage of the present invention is characterized in that the thickness of the taste is imparted even though the sugar concentration is reduced. Here, the "taste thickness" means a flavor sensation recognized by the spread, complexity, body feeling, and the like of the taste.

The beer-taste fermented alcoholic beverage of the present invention is a normal beer as long as the sugar 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 produced according to the production procedure of the taste fermented alcoholic beverage.

For example, fermented beer yeast is added to wort prepared from brewing raw materials such as malt, hops, auxiliary raw materials, and brewing water to ferment, and the obtained fermented liquid is stored at a low temperature and then filtered. By removing yeast, a beer-taste fermented alcoholic beverage can be produced. It goes without saying that among beer-taste fermented alcoholic beverages, all-malt beer can be produced from malt, hops, and water.

In the above production procedure, wort can be produced according to a conventional method. For example, a mixture of brewing ingredients and brewing water is saccharified and filtered to obtain wort, hops are added to the wort, and then the wort is boiled and the boiled wort is cooled to prepare the wort. Can be done. The wort can also be prepared by adding a commercially available enzyme preparation during the saccharification step. For example, a protease preparation for proteolysis, an α-amylase preparation, a glucoamylase preparation, a plulanase preparation, etc. for sugar decomposition, a β-glucanase preparation, a fibrinolytic enzyme preparation, etc. for fibrinolytic decomposition, respectively. It can be used, or a mixed preparation of these can be used.

In one aspect of the production of the beer-taste fermented alcoholic beverage of the present invention, in the saccharification step or fermentation step, malt having high endoprotease activity and / or an enzyme preparation having endoprotease activity is used for proteolysis. be able to. By carrying out such a step, the ratio of peptides having a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) contained in the beer-taste fermented alcoholic beverage can be increased and adjusted within a predetermined value range. As a result, it is possible to produce a beer-taste fermented alcoholic beverage having a thick taste while having no sugar. That is, according to the present invention, there is provided a method for producing a beer-taste beverage, which comprises using an enzyme preparation containing malt and / or end-type protease having high endo-type protease activity in a saccharification step or fermentation step. The characteristics of the enzyme activity of the above malt and enzyme preparations can be identified based on the endo-type protease activity.

The beer-taste fermented alcoholic beverage of the present invention is one in which either or both of malt and ungerminated wheat are used as raw materials, and preferably at least malt or malt and ungerminated wheat are used as raw materials. It is a part. In the beer-taste fermented alcoholic beverage of the present invention, barley malt can be used as a part of the raw material as malt. In the beer-taste fermented alcoholic beverage of the present invention, ungerminated barley (including extracted one) and ungerminated wheat (including extracted one) are used as raw materials as ungerminated wheat. Can be done.

In the production of the beer-taste fermented alcoholic beverage of the present invention, in addition to malt and ungerminated wheat, rice, corn, soybean, koryan, potato, starch, sugar (for example, liquid sugar), fruit (for example, fruit juice, concentrated). Juice), coriander or its seeds, spices or their ingredients (eg pepper, cinnamon, sansho), herbs (eg chamomile, sage, basil), vegetables (eg kansho, pumpkin), buckwheat or sesame, sugar-containing substances Auxiliary ingredients such as (eg, honey, brown sugar), salt, miso, flowers, tea, coffee, cocoa (tea, coffee and cocoa include their preparations), marine products (eg, oysters, konbu, wakame, katsuo-bushi); Nitrogen sources such as proteolytic products and yeast extracts; other additives such as fragrances, pigments, foaming / foaming improvers, water conditioners, fermentation aids and the like can be used as brewing ingredients. In the beer-taste fermented alcoholic beverage of the present invention, the raw materials used other than the water for brewing can be at least malt and hops, and in some cases, sugars, rice, corn, starch and the like can be used as raw materials.

In the present invention, a fraction containing a peptide having a molecular weight of 800 to 1500 Da is prepared from grains (for example, malt and / or ungerminated wheat or soybean), and the fraction is used as a sugar-free beer-taste beverage. By adding it, the ratio of peptides having a molecular weight of 800 to 1500 Da can be adjusted within a predetermined value range, and by extension, a beer-taste fermented alcoholic beverage having zero sugar and a thickness of taste can be produced. Can be done. That is, according to the present invention, a fraction containing a peptide having a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) is prepared from grains, and the fraction is used as a beer having a sugar concentration of less than 0.5 g / 100 mL. A method for producing a beer-taste fermented alcoholic beverage is provided, which comprises adding to the taste-fermented alcoholic beverage. The peptide fraction can be added to a beer-taste fermented alcoholic beverage in a manufacturing process of the beverage (for example, a step of blending into a fermented liquid). A peptide having 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 hydrolyzing soybean protein. It may be obtained from a protein hydrolyzate of grains such as stuff.

In the present invention, a beer-taste fermented alcoholic beverage using malt and / or ungerminated wheat as a raw material is produced, and a fraction containing a peptide having a molecular weight of 800 to 1500 Da is prepared from the beverage. By adding the fraction to a sugar-free beer-taste beverage, the ratio of peptides having a molecular weight of 800 to 1500 Da can be adjusted within a predetermined value range, and as a result, the thickness of the taste can be adjusted while having zero sugar. An imparted beer-taste fermented alcoholic beverage can be produced. That is, according to the present invention, a beer-taste fermented alcoholic beverage using malt and / or ungerminated wheat as a raw material is produced, and then a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) is produced from the beverage. A method for producing a beer-taste fermented alcoholic beverage, which comprises preparing a fraction containing the peptide of) and adding the fraction to a beer-taste fermented alcoholic beverage having a sugar concentration of less than 0.5 g / 100 mL. Provided. The peptide fraction can be added to a beer-taste fermented alcoholic beverage in a manufacturing process of the beverage (for example, a step of blending into a fermented liquid).

Further, according to the present invention, one or more peptides having a molecular weight of 800 to 1500 Da derived from a beer-taste fermented alcoholic beverage derived from grains or malt and / or ungerminated barley as a raw material. A beer-taste fermented alcoholic beverage that is compounded is provided, and the beverage is a part of the beer-taste fermented alcoholic beverage of the present invention. The beverage containing the peptide has an improved or improved flavor as a beer-taste fermented alcoholic beverage, and specifically, a beverage having an enhanced taste thickness.

The reduction of carbohydrates in beer-taste fermented alcoholic beverages can be performed according to known methods, for example, a method of adding glucoamylase during the saccharification step, a method of adding glucoamylase during the fermentation step (enzyme utilization technology system basics)・ From analysis to modification, advanced functionality, and industrial use (see Makoto Komiyama, NTS Co., Ltd., 2010), yeast uses liquid sugar containing a large amount of sugar assimilated into yeast. Sugar in beverages by a method of reducing sugar by increasing the assimilation property (see JP-A-2009-131202) and a method of filtering wort during the saccharification step (see JP-A-2012-147780). The quality concentration can be set to a predetermined value.

In some cases, the beverage provided by the present invention can be provided as a packaged beverage through a step of adding carbon dioxide gas, a filling step, a sterilization step, and the like. Sterilization may be performed before or after filling the container. Further, when the pH of the beverage is adjusted to less than 4, the filling step can be performed as it is without going through the sterilization step to obtain a packaged beverage.

The container used for the beverage according to the present invention may be a container normally used for filling the beverage, for example, a metal can, a barrel container, a plastic bottle (for example, PET bottle, a cup), a paper container, a bottle, etc. Examples thereof include pouch containers, but metal cans / barrel containers, plastic bottles (for example, PET bottles), and bottles are preferable.

According to another aspect of the present invention, there is a method for producing a sugar-free beer-taste fermented alcoholic beverage using at least a part of malt and / or ungerminated wheat as a raw material with improved flavor. The ratio (percentage) of the mass of the peptide having a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the beverage to the mass of all the peptides in the beverage fractionated by the gel filtration method for HPLC analysis is 17 to 45%. Methods are provided that include adjusting. In the present invention, "improved flavor" or "improved flavor" of a beer-taste fermented alcoholic beverage means that the thickness of the taste is realized in a beer-taste fermented alcoholic beverage having no sugar. The above-mentioned production method of the present invention is preferably a method for producing a sugar-free beer-taste fermented alcoholic beverage having a thickness of taste. The above-mentioned production method of the present invention can be carried out in accordance with the description of the beer-taste fermented alcoholic beverage of the present invention and the production method thereof.

According to another aspect of the present invention, a beer-taste fermented alcoholic beverage containing grains or malt and / or ungerminated wheat as a raw material has a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis). A sugar-free beer-taste fermented alcoholic beverage flavor-improving agent containing the above peptide as an active ingredient is provided. The flavor improving agent for beer-taste fermented alcoholic beverages is preferably a sugar-free beer-taste fermented alcoholic beverage flavor-imparting agent. The flavor improving agent of the present invention can be carried out in accordance with the description regarding the beer-taste fermented alcoholic beverage of the present invention and the method for producing the same.

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 using malt and / or ungerminated wheat as a part of the raw material. In the flavor improving method of the present invention, the ratio of the mass of peptides having a molecular weight of 800 to 1500 Da (the gel filtration method for HPLC analysis) in the beverage to the mass of all the peptides in the beverage fractionated by the gel filtration method for HPLC analysis ( This can be done by adjusting the percentage) to 17-45%. The flavor improving method of the present invention is preferably a method of imparting a thickness of taste to a sugar-free beer-taste fermented alcoholic beverage. The flavor improving method of the present invention can be carried out in accordance with 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.

Analysis of carbohydrate concentration The measurement of carbohydrate concentration based on the nutrition labeling standard was performed according to the method described in the known 5th edition Standard Tables of Food Composition in Japan Analysis Manual. That is, from the mass of the sample beverage, water content (vacuum heating and drying method), protein (method by automatic analyzer), lipid (soxley extraction method (3)), ash content (combustion method by muffle furnace) and amount of dietary fiber (proski). It was measured by a method calculated excluding the enzyme gravimetric method).

Reference Example 1: Identification of a peptide fraction that imparts a favorable flavor to beer-taste beverages (1) Production of beer-taste fermented alcoholic beverages Beer-taste fermented alcoholic beverages are prepared by the infusion method using barley malt, hops, and enzyme preparations. Manufactured. Specifically, 100 g of barley malt was placed in 300 mL of hot water at 65 ° C., held for 60 minutes, further heated to 78 ° C. and 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, and then filtered to obtain a pre-fermentation liquid.

Then, fermentation was carried out with brewer's yeast according to a conventional method, and the flavor of the fermented liquid was confirmed. As a result, it was confirmed that the obtained fermented liquid had a soft and smooth texture like beer with suppressed miscellaneous taste and had a harmonious taste.

(2) Gel Filtration Fraction The fermented liquid obtained in (1) above was filtered through a 0.45 μm filter, the filtered fermented liquid was weighed, and freeze-dried. The dried product was dissolved in 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 fraction under the following conditions.

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

According to the sensory evaluation of the fractions, the fractions were divided into 9 groups of pre-fractions, A1, A2, B, C, D, E, F and G as shown in Table 1 according to the difference in flavor characteristics. In addition, the flavor characteristics of each fraction were determined by discussions with five trained panelists.

(3) Purification of Peptide Fractions Fractions C and D obtained in (2) above are adsorbed on a solid-phase extraction column (Bond Elute C18, manufactured by Agilent technologies) and washed with desalinated water. , Elution with 50% aqueous ethanol solution, concentrated to dryness, and reconstituted with desalted water to prepare a concentrated solution. This was used as a peptide fraction.

(4) Protein quantification by the Lowry method The protein quantification of the peptide fraction obtained by the gel filtration fraction of the above (2) and the purification of the peptide fraction of the above (3) is a commercially available kit (DC protein assay, Bio-. The Lowry method using (manufactured by Rad) was used. First, the concentration of the fractionated solution was adjusted so as to be within an appropriate range. To 5 μL of the sample whose concentration was adjusted, 50 μL of solution A was added and stirred, and then 400 μL of solution B was added and stirred. After the color reaction was carried out at room temperature for 15 minutes, 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 the calibration curve prepared in advance. The calibration curve was prepared using BSA (bovine serum albumin).

(5) Results The peptide fraction was analyzed on a Superdex 75 10/300 column to estimate the molecular weight. As a result, the peptides of the fractions C and D were distributed at a molecular weight of about 800 to 1500 Da.

From the above results, it was suggested that the peptide fractions C and D having a molecular weight of about 800 to 1500 Da, which were fractionated and purified from the beer-taste fermented alcoholic beverage, were effective in imparting a drinking response (that is, taste thickness). ..

Example 1: Production of zero-sugar beer-taste fermented alcoholic beverage and analysis of the effect of addition of peptides to the beverage on the thickness of taste (1) Production of zero-sugar beer-taste fermented alcoholic beverage Beer-taste fermentation of this example In the production of alcoholic beverages, barley malt was used as the main raw material. For saccharification, an enzyme preparation was used, the temperature and time of saccharification were adjusted, and the mixture was filtered to obtain wort. Specifically, 10 parts by mass of barley malt, an enzyme preparation containing β-glucanase and an enzyme preparation mainly composed of glucoamylase are added to 100 parts by mass of hot water at 50 ° C., and the temperature is raised to 64 ° C. after holding for 30 minutes. It was held for 70 minutes. Then, the temperature was raised to 78 ° C. and the temperature was maintained for 5 minutes, and then the mixture was filtered to obtain wort.

Following the above wort preparation step, 9.5 parts by mass of liquid sugar mainly composed of hops and assimilating sugar is added to the obtained wort and boiled at 100 ° C. for 90 minutes, and then the wort is allowed to stand. After separating the trube, it was cooled to obtain a pre-fermentation wort. Then, bottom fermentation yeast was added to the pre-fermentation liquid, and main fermentation and post-fermentation were carried out according to a conventional method. Subsequently, the fermented liquor after post-fermentation was stored 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. The lyophilized product was dissolved in a 100 mM NaCl solution to prepare a 5-fold concentrated solution, which was used as a sample for gel filtration fractionation. A gel filtration fraction was carried out according to the description of Reference Example 1 (2), and the obtained fraction C (fraction number: F25 to 29) was purified according to the description of Reference Example 1 (3). At this time, in order to improve the recovery rate in purification, the passing fraction of the solid-phase extraction column was also recovered with a synthetic adsorbent (Sepa beads 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 brewed product, the sample brewed product obtained in (1) above was degassed, weighed, and freeze-dried. Each lyophilized product was dissolved in 50 mM sodium phosphate buffer (containing pH 7.0 and 150 mM NaCl) 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 solution so as to be equivalent to a 10-fold concentrated solution, and used as a sample for gel filtration fraction. Each of the fractionated samples was subjected to gel filtration fractionation under the following conditions to obtain fractionation fractions (fractions 1 to 10).

The conditions of the gel filtration method for HPLC analysis were as follows.
<Analytical conditions for gel filtration method for HPLC analysis>
Column: Superdex 75 10/300 (manufactured by GE Healthcare)
Sample injection volume: 100 μL
Eluent composition: 50 mM sodium phosphate (pH 7.0), 20% (v / v) acetonitrile, 150 mM NaCl
Flow velocity: 0.5 mL / min (constant flow velocity)
Detection wavelength: 215 nm

The protein quantification of the fractionation solution was performed by the Lowry method described in Reference Example 1 (4).

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

B. Preparation of molecular weight calibration line In the column, eluent, flow velocity, and detection wavelength described above for HPLC analysis gel filtration conditions, 50 μL of a peptide having a known molecular weight dissolved in ultrapure water at 0.1 to 5 mg / mL was injected. Then, HPLC analysis was performed to confirm the retention time (Table 2). A calibration curve (Fig. 1) was prepared from the holding time and molecular weight.

C. Calculation of molecular weight As a result of molecular weight measurement, it was confirmed that the peptides contained in fraction 7 out of fractions 1 to 10 have a molecular weight in the range of 800 to 1500 Da.

(5) Calculation of Peptide Ratio with Molecular Weight of 800 to 1500 Da The peptide ratio with a molecular weight of 800 to 1500 Da was calculated by the following formula using the peptide concentration corresponding to the product in each fractionation fraction.

The brewed peptide ratios were as shown in Table 3.

(6) Analysis of sugar concentration and original extract concentration The sugar concentration of the sample was measured based on the nutrition labeling standard as described above. The results were as shown in Table 3. The original extract concentration (OE concentration) of the sample brewed product was measured according to the BCOJ beer analysis method (Beer Sake Brewing Association (2013), 8.3.6, 8.4.3 and 8.5. The measurement was performed. This was performed using an alcoholizer (manufactured by Anton Pearl).

(7) Sensory evaluation A test in which the peptide fraction obtained in (2) above is not added to the beer-taste fermented alcoholic beverage (malt usage ratio: 50%) produced in (1) above, and a test in which it is added. Sections (additives 1 to 9) were provided, and sample beverages (sample numbers 1 to 10) having a peptide concentration of 800 to 1500 Da adjusted as shown in Table 3 were prepared.

Sensory evaluation was performed by 5 trained panelists. Specifically, "taste thickness" and "miscellaneous taste" were evaluated on a scale of 1 to 9 in 0.5-point increments, and the average value of the evaluation scores of the five panelists was calculated. Here, the "taste thickness" refers to a flavor sensation recognized by the spread, complexity, body feeling, and the like of the taste. Regarding the "taste thickness", the sample beverage of sample number 1 is 1 point (there is no taste thickness), the sample beverage of sample number 6 is 5 points (there is a taste thickness), and 1.5 points or more is the taste. It was judged that the thickness was improved. In addition, "miscellaneous taste" means astringent taste, astringent taste, and astringent taste. Regarding "miscellaneous taste", the sample beverage of sample number 1 was 1 point (no miscellaneous taste), the sample beverage of sample number 6 was 3 points (some astringency), and 4.0 points or more was said to have improved the astringency. It was judged.

Based on the sensory evaluation results for "thickness of taste" and "presence or absence of miscellaneous taste", a comprehensive evaluation was performed according to the following criteria.
⊚: The thickness of the taste is improved (2.5 points or more), and the taste is less (less than 4.0 points).
◯: The thickness of the taste is slightly improved (1.5 points or more) and the unpleasant taste is small (less than 4.0 points), or the thickness of the taste is improved (2.5 points or more) but the unpleasant taste is present. (4.0 points or more)
×: The thickness of the taste is not improved (less than 1.5 points)

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

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

Claims (6)

A beer-taste fermented alcoholic beverage containing malt as at least a part of the raw material and having a sugar concentration of less than 0.5 g / 100 mL, based on the total amount of peptides in the beverage fractionated by the gel filtration method for HPLC analysis. A beer taste in which the ratio (percentage) of the mass of the peptide having a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the beverage is 18 to 25 %, and the peptide uses malt as a raw material at least. A beer-taste fermented alcoholic beverage derived from a fermented alcoholic beverage. The beer-taste fermented alcoholic beverage according to claim 1, wherein the malt usage ratio is 50% or more. A method for producing a beer-taste fermented alcoholic beverage having an improved flavor, using malt as at least a part of the raw material, and having a sugar concentration of less than 0.5 g / 100 mL, which is fractionated by a gel filtration method for HPLC analysis. The ratio (percentage) of the mass of the peptide having a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the beverage to the mass of the total peptide of the beverage was adjusted to 18 to 25 %. A method in which the peptide is derived from a beer-taste fermented alcoholic beverage that uses malt as a raw material. A claim including a step of adding a peptide having a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) contained in a beer-taste fermented alcoholic beverage containing at least a part of malt as a raw material in the process of producing a beer-taste fermented alcoholic beverage. Item 3. The manufacturing method according to Item 3. A sugar concentration of less than 0.5 g / 100 mL containing a peptide having a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) derived from a beer-taste fermented alcoholic beverage using malt as a raw material as an active ingredient. Beer taste Fermented alcoholic beverage flavor improver. A method for improving the flavor of a beer-taste fermented alcoholic beverage having a sugar concentration of less than 0.5 g / 100 mL and using malt as a raw material, and all of the beverages fractionated by the gel filtration method for HPLC analysis. It comprises adjusting the ratio (percentage) of the mass of the peptide in the beverage to the mass of the peptide having a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) to 18 to 25 %, and the peptide is fermented . A method derived from a beer-taste fermented alcoholic beverage that uses at least part of the raw material.

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