JP6785102B2 - Low-carb beer taste beverage - Google Patents

Description

The present invention relates to a low-carbohydrate beer-taste beverage, and more particularly to a beer-taste beverage having a sugar concentration of less than 1.1 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 technologies 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

Low-carb beer-taste beverages have a watery, light-flavored impression due to the reduced sugar content. Therefore, if the thickness of the taste can be realized in a low-carbohydrate beer-taste beverage, it is possible to provide a low-carbohydrate beer-taste beverage that is responsive to drinking.

That is, an object of the present invention is to provide a low-carbohydrate beer-taste beverage in which the thickness of the taste is realized.

The present inventors have found that in a beer-taste beverage having a reduced sugar concentration, the thickness of the taste can be realized in the beer-taste beverage by setting the ratio of peptides having a molecular weight of 800 to 1500 Da within a specific range. The present invention is based on these findings.

According to the present invention, the following inventions are provided.
[1] A beer-taste beverage containing malt and / or ungerminated wheat as at least a part of the raw material and having a sugar concentration of less than 1.1 g / 100 mL, which was fractionated by a gel filtration method for HPLC analysis. A beer-taste beverage in which the ratio (percentage) of the peptide mass of a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the beverage to the total mass of the peptide derived from the beverage is 25 to 45%.
[2] The beer-taste beverage according to the above [1], wherein the ratio of the peptide mass is 28 to 38%.
[3] The beer-taste beverage according to the above [1] or [2], which is a fermented alcoholic beverage.
[4] A method for producing a beer-taste beverage having an improved flavor, using malt and / or ungerminated wheat as a raw material and having a sugar concentration of less than 1.1 g / 100 mL, which is analyzed by HPLC. The ratio (percentage) of the molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the beverage to the total mass of the peptide derived from the beverage fractionated by the gel filtration method for HPLC analysis is adjusted to 25 to 45%. How to include.
[5] 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 beverages with a sugar concentration of less than 1.1 g / 100 mL.
[6] A method for improving the flavor of a beer-taste beverage having a sugar concentration of less than 1.1 g / 100 mL, wherein the molecular weight in the beverage is 800 to the total mass of peptides derived from the beverage fractionated by the gel filtration method for HPLC analysis. A method comprising adjusting the peptide mass ratio (percentage) of 1500 Da (gel filtration for HPLC analysis) to 25-45%.

According to the present invention, in a beer-taste fermented alcoholic beverage having a sugar concentration of less than 1.1 g / 100 mL, a low sugar having a thickness of taste is realized by adjusting the peptide ratio 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 and can meet various needs such as consumer health consciousness.

It is a figure which showed the peptide amount of the beer taste fermented alcoholic beverage to which the peptide (fractions C and D) was added in Reference Example 1. It is a figure which showed the sensory evaluation result of Reference Example 1. FIG. Sensory evaluation scores were shown for fractions C and D. The sensory evaluation score of the additive-free control was 2.5. It is a figure which showed the sensory evaluation result of the sample brewed product and the peptide additive product produced in Example 1. 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 2. FIG.

Specific description of the invention

In the present invention, the "beer-taste beverage" means a beverage having a beer-specific taste and aroma obtained when beer is normally produced, that is, when beer is produced based on fermentation with yeast or the like, and "beer". Includes "taste alcoholic beverages" and "beer-taste non-alcoholic beverages."

"Beer-taste alcoholic beverages" also include "beer-taste fermented alcoholic beverages" that are fermented with yeast using carbon sources, nitrogen sources, water, etc. as raw materials, and "beer-taste fermented alcoholic beverages" include beer. , Happoshu and beer that uses malt as a raw material, and beverages made by adding alcohol to happoshu (for example, liqueur-based new genre beverages classified as "liqueur (foaming) (1)" under the Liquor Tax Law) Can be mentioned.

The beer-taste 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 less than 50%. , 50% or more, less than 67%, or 67% 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 non-alcoholic beverage" is a non-alcoholic beverage that does not contain alcohol components derived from fermentation because it is unfermented, but has a beer-like flavor. Further, in the present invention, the "non-alcoholic beverage" means a beverage containing no alcohol, that is, an ethanol concentration of 0.00v / v%, and is synonymous with a "completely alcohol-free beverage".

The beer-taste beverage of the present invention is a low-carbohydrate beverage having a reduced sugar concentration. Specifically, the beer-taste beverage of the present invention has a sugar concentration of less than 1.1 g / 100 mL. The sugar concentration of the beer-taste fermented alcoholic beverage of the present invention can be preferably 1.0 g / 100 mL or less.

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 beverage of the present invention, the ratio (percentage) of the peptide mass of the molecular weight 800 to 1500 Da (gel filtration method for HPLC analysis) in the beverage to the total peptide mass derived from the beverage fractionated by the gel filtration method for HPLC analysis is It is characterized by being within a specific range. In the present invention, the peptide ratio ranges from 25 to 45%, preferably 28 to 38%.

The molecular weight of a peptide in a beverage is measured by a gel filtration method by high performance liquid chromatography (sometimes referred to as "gel filtration method for HPLC analysis" in the present specification). 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. 4), 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 2 below. Further, the quantification of the peptide can be carried out by the Lowry method (Lowry method).

The beer-taste 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 beverage of the present invention is a normal beer-taste beverage 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 manufactured according to the manufacturing procedure of.

For example, when the beverage of the present invention is a beer-taste alcoholic beverage, fermentation is carried out by adding fermentation beer yeast to wort prepared from brewing raw materials such as malt, hops, auxiliary raw materials, and brewing water. A beer-taste fermented alcoholic beverage can be produced by storing the fermented wort at a low temperature and then removing yeast by a filtration step. 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. Wort can also be prepared by adding a commercially available enzyme preparation during the saccharification step. For example, protease preparations for proteolysis, α-amylase preparations, glucoamylase preparations, pullulanase preparations, etc. for carbohydrate decomposition, β-glucanase preparations, fibrinolytic enzyme preparations, etc. for fibrinolytic decomposition, respectively. It can be used, or a mixed preparation of these can be used.

In a preferred embodiment of the production of the beer-taste beverage of the present invention, an enzyme preparation having end-type protease activity can be used for proteolysis in a saccharification step or a fermentation step. 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 beer-taste beverages can be increased and adjusted within a predetermined value range, and thus low sugar. It is possible to produce a beer-taste beverage that is of high quality but has a thick taste. 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 an endotype protease for proteolysis in a saccharification step or a fermentation step.

The activity characteristics of the enzyme preparation used for the above proteolysis can be identified based on the endo-type protease activity, for example, the enzyme preparation is used so that the end-type protease activity value is 284 to 8000 units / kg of malt. can do. More preferably, the enzyme preparation can be used so that the endo-type protease activity value is 635 to 4000 units / kg of malt. The activity measurement of the end-type protease activity can be performed according to the procedure described in Example 2.

In the production of the beer-taste alcoholic beverage of the present invention, in addition to malt, ungerminated barley (for example, ungerminated barley (including extracted one), ungerminated wheat (including extracted one)); rice, Auxiliary raw materials stipulated by the Liquor Tax Law such as corn, koryan, malt, starch, sugar (for example, liquid sugar); nitrogen sources such as proteolytic products and yeast extract; Other additives such as agents and fermentation aids can be used as brewing ingredients. In the beer-taste alcoholic beverage of the present invention, at least malt and hops can be used as raw materials other than brewing water, and in some cases, sugars, rice, corn, starch and the like can be used as raw materials. It goes without saying that among beer-taste fermented alcoholic beverages, all-malt beer can be produced from malt, hops, and water.

For beer-taste non-alcoholic beverages, malt, hops, other raw materials (for example, fragrances, pigments, dietary fibers, protein hydrolysates, foaming / foam retention improvers, water quality modifiers, etc.), water and other raw materials A beer-taste non-alcoholic beverage can be produced by preparing a preparation liquid from the preparation liquid, removing solid content from the preparation liquid by standing, and then filtering the preparation liquid. The above-mentioned preparation liquid can be prepared according to a conventional method in the same manner as the above-mentioned method for preparing wort for a beer-taste fermented alcoholic beverage. In the saccharification step or fermentation step, an enzyme preparation containing an endoprotease can be used for proteolysis.

In the present invention, a beer-taste fermented alcoholic beverage using malt and / or ungerminated wheat as a raw material is produced, and a peptide containing a peptide having a molecular weight of 800 to 1500 Da is contained in the beverage according to the procedure of Example 1. By preparing the portion and adding the fraction to a low-sugar 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 by extension, the taste is low-sugar. It is possible to produce a beer-taste beverage with the thickness of. 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. ) Is prepared, and a method for producing a beer-taste beverage is provided, which comprises adding the fraction to a beer-taste beverage having a sugar concentration of less than 1.1 g / 100 mL.

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 using malt and / or ungerminated wheat as a raw material are blended. A beer-taste beverage is provided, which is part of the beer-taste beverage of the present invention. The beverage containing the peptide has an improved or improved flavor as a beer-taste beverage, and specifically, a beverage having an enhanced taste thickness.

Carbohydrate reduction in beer-taste beverages can be performed 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 / analysis). From to modification / advanced functionalization / industrial use (see Makoto Komiyama supervision, NTS Co., Ltd., 2010), using liquid sugar containing a large amount of sugar assimilated into yeast. Concentration of carbohydrates in a beverage by a method of reducing carbohydrates by increasing the saccharification property (see JP-A-2009-131202) and a method of filtering wort during the saccharification step (see JP-A-2012-147780). 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 usually 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, a method for producing a low-sugar beer-taste beverage using malt and / or ungerminated wheat as a raw material with improved flavor, for HPLC analysis. Including adjusting the ratio (percentage) of the molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) of the molecular weight in the beverage to the total mass of the peptides derived from the beverage fractionated by the gel filtration method to 25 to 45%. A method is provided. In the present invention, "improved flavor" or "improved flavor" of a beer-taste beverage means that the thickness of the taste is realized in a low-carbohydrate beer-taste beverage. The above-mentioned production method of the present invention is preferably a method for producing a low-carbohydrate beer-taste beverage to which a thickness of taste is imparted. The above-mentioned production method of the present invention can be carried out in accordance with the description of the beer-taste beverage of the present invention and the production method thereof.

According to another aspect of the present invention, 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. A flavor improving agent for a low sugar beer-taste beverage, which is contained as an active ingredient, is provided. The flavor improving agent for beer-taste beverages is preferably a taste thickness-imparting agent for low-carbohydrate beer-taste beverages. The flavor improving agent of the present invention can be carried out in accordance with the description regarding the beer-taste 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 low-carbohydrate beer-taste 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 (percentage) of the peptide mass having a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) in the beverage to the total peptide mass derived from the beverage fractionated by the gel filtration method for HPLC analysis. Can be implemented by adjusting to 25-45%. The flavor improving method of the present invention is preferably a method of imparting a thickness of taste to a low-carbohydrate beer-taste beverage. The flavor improving method of the present invention can be carried out in accordance with the description of the beer-taste 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 (Soxhlet 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 solution.

Then, fermentation was carried out with brewer's yeast according to a conventional method, and the flavor of the fermented liquid was confirmed. Eight trained panelists have "soft taste", that is, the minimum is one point, the highest is based on the fact that it has a soft and smooth texture like beer, and has a harmonious taste. Was evaluated as 5 points on a 5-point scale, and the average score was calculated. The results were as shown in Table 1.

As shown in Table 1, the produced beer-taste fermented alcoholic beverages had a good sensory evaluation score (softness of taste) and a favorable impression of flavor.

(2) Gel Filtration Fraction The fermentation broth obtained in (1) above was filtered through a 0.45 μm filter, the filtered fermented broth 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 5 mL fractions 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 2 according to the difference in flavor characteristics.

(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 desalinated 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 (3) above was performed by the Lowry method using a commercially available kit (DC protein assay, manufactured by Bio-Rad). .. 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) Sensory Evaluation These fractions / refined samples were used in a commercially available fermented alcoholic beverage with a beer taste using barley and barley malt with a malt usage ratio of less than 49%, and the fractions C and D contained in the beverage were used. Fractions C and D were added so as to add 50% of the amount (see FIG. 1), and sensory evaluation was performed by 5 trained panelists.

The index of sensory evaluation was evaluated on a five-point scale of 1 to 5 as a comprehensive evaluation of "umami, sweetness, thickness, body, and astringency / taste incongruity as off-flavour". The additive-free control was given a score of 2.5. The average value of the sensory evaluation score was as shown in FIG. 2, and the sensory evaluation comment was as shown in Table 3.

As shown in FIG. 2, when the fractions C and D were added, the sensory evaluation score was high, which was in agreement with the sensory evaluation result of the fermented liquid before the fraction. In addition to the high scores of fractions C and D, the taste was evaluated in the sensory evaluation comments, and in discussions with the panelists, it was evaluated that the thickness and response of the taste and the contribution to the body were strong. It was. That is, it was found that the fractions C and D have an effect of contributing to the thickness, the body and the like, and the umami.

The peptide fraction was analyzed on a Superdex 75 10/300 column and the molecular weight was estimated. 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 clarified that the fractionated and purified peptide fractions C and D having a molecular weight of about 800 to 1500 Da can impart thickness, body and umami to beer-based beverages.

Example 1: Production of low-sugar beer-taste fermented alcoholic beverage and analysis of the effect of peptide addition on the beverage on the thickness of taste (1) Production of low-sugar beer-taste fermented alcoholic beverage by pilot plant Beer taste of this example In the production of fermented alcoholic beverages, barley malt was used as the main raw material. At the time of saccharification, an enzyme preparation was used, the temperature and time of saccharification were adjusted, and the mixture was filtered to obtain wort. Specifically, 33.6 parts by mass of barley malt and a β-glucanase enzyme preparation were added to 100 parts by mass of hot water at 64 ° C. and held for 100 minutes, then heated to 78 ° C. and held for 5 minutes. It was filtered to obtain wort.

Following the above wort preparation step, hops are added to the obtained wort and boiled at 100 ° C. for 90 minutes, the wort is allowed to stand, the trube is separated, and then cooled to prepare the pre-fermentation solution. Obtained. An enzyme preparation mainly containing glucoamylase was added to the obtained pre-fermentation solution. 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 all-malt type low-carbohydrate beer-taste alcoholic beverage.

(2) Preparation of peptide fraction (a) Gel filtration fraction All-malt beer (commercially available) 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. The sample was subjected to gel filtration fractionation under the same conditions as the gel filtration fractionation condition of Reference Example 1. Further, as in Reference Example 1, the fractionated fractions were divided into nine groups shown in Table 2.

(B) Purification of peptide fractions The fractions C to D (fraction numbers: F25 to 33) obtained in (a) above are purified peptides in the same manner as the purification of the peptide fraction described in Reference Example 1. I got a fraction.

(C) Protein quantification by the Lowry method The peptide fraction obtained by the gel filtration fraction of (a) above is added to the low-sugar beer-taste fermented alcoholic beverage produced in (1) above, and the HPLC according to Example 2 Gel filtration fractionation was performed by the analytical gel filtration method. The protein quantification of the peptide fraction obtained thereby was adjusted in concentration so as to be within an appropriate range, 25 μL of solution A was added to 5 μL of the sample and stirred, and then 200 μL of solution B was added, and described in Reference Example 1. The same procedure as for protein quantification was performed.

(3) Calculation of Peptide Ratio of Molecular Weight of 800 to 1500 Da The peptide ratio of molecular weight of 800 to 1500 Da was calculated by the following formula.

(4) Analysis of carbohydrate concentration The carbohydrate concentration of the sample brewed product and the additive (Sample Nos. 1 to 3) based on the nutrition labeling standard was performed as described above. The results were as shown in Table 4.

(5) Sensory evaluation A test group (trial product) in which the peptide fraction obtained in (2) above is not added and a test group (additive product (additive product)) in which the peptide fraction obtained in (2) above is not added to the low-sugar beer taste fermented alcoholic beverage produced in (1) above. 1) and additive (2)) were provided, and sample beverages (sample numbers 1 to 3) having a peptide concentration of 800 to 1500 Da adjusted as shown in Table 4 were prepared.

Sensory evaluation was performed by 5 trained panelists. Specifically, the "taste thickness" was evaluated on a 9-point scale of 1 to 9, 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 was given 3 points (there was no taste thickness), and those having a higher numerical value than sample number 1 were judged to have increased taste thickness.

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

From the results of Table 4 and FIG. 3, even in the case of a beer-taste fermented alcoholic beverage having a sugar concentration of less than 1.1 g / 100 mL, the "taste thickness" can be improved by increasing the peptide ratio of 800 to 1500 Da. confirmed.

Example 2: Preparation of a low-sugar beer-taste fermented alcoholic beverage and analysis of the effect of a peptide having a molecular weight of 800 to 1500 Da on the thickness and miscellaneous taste (1) Production of a low-sugar beer-taste fermented alcoholic beverage by a pilot plant This Example In the production of beer-taste fermented alcoholic beverages, barley malt was used as the main raw material. At the time of saccharification, an enzyme preparation was used, the temperature and time of saccharification were adjusted, and the mixture was filtered to obtain wort. Specifically, 33.6 parts by mass of barley malt and a β-glucanase enzyme preparation were added to 100 parts by mass of hot water at 64 ° C. and held for 100 minutes, then heated to 78 ° C. and held for 5 minutes. It was filtered to obtain wort.

Following the above wort preparation step, hops are added to the obtained wort and boiled at 100 ° C. for 90 minutes, the wort is allowed to stand, the trube is separated, and then cooled to prepare the pre-fermentation solution. Obtained. An enzyme preparation mainly containing glucoamylase was added to the obtained pre-fermentation solution. 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 all-malt type beer-taste fermented alcoholic beverage (sample numbers 11 to 15). Further, sample number 14 and sample number 15 were mixed at a ratio of 3: 2 to obtain a beer-taste fermented alcoholic beverage (sample number 16).

(2) Protease activity in enzyme preparation The external enzyme preparation used in (1) above is an enzyme preparation containing an endo-type protease and can be specified based on the endo-type protease activity. Specifically, it was added to each test group so that the endo-type protease activity value was 50 to 7000 units / kg of malt.

The activity of the end-type protease was measured by using milk casein (pH 6) as an enzyme reaction substrate, performing an enzyme reaction, and measuring the amino acids released in the solution. The specific measurement method is as follows.

A 13.3 mM sodium phosphate buffer (pH 6.0) containing 0.6% milk casein was used as a milk casein substrate. The enzyme preparation was appropriately diluted with ion-exchanged water. To 5 mL of the milk casein substrate solution, 1 mL of the enzyme preparation diluted solution was added, and the reaction was carried out at 30 ° C. for 10 minutes. After completion of the reaction, 5 mL of 0.44 M trichloroacetic acid solution was added, mixed well, and left at 30 ° C. for 30 minutes to stop the reaction and insolubilize the undecomposed substrate. No. The reaction solution was filtered through 6 filter papers. 2.5 mL of 0.55 M sodium carbonate solution was added to 1 mL of the filtrate, 0.5 mL of a 3-fold diluted phenol reagent was added, and the mixture was left at 30 ° C. for 30 minutes to allow sufficient color development. The absorbance at 660 nm was measured with an absorbance meter (manufactured by Shimadzu Corporation), and the amino acids released from the substrate were quantified. Enzyme activity was defined as 1 unit (Unit) of the amount of enzyme that resulted in an increase in the Folin reagent colorant corresponding to 1 μg of tyrosine per minute.

(3) Preparation of Gel Filtration Fraction for HPLC Analysis The sample brewed products (sample numbers 11 to 16) obtained in (1) above were 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. Each sample was subjected to gel filtration fractionation under the following conditions.

The conditions of the gel filtration method for HPLC analysis were as follows.
<Analysis 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 carried out according to the Lowry method as in Reference Example 1.

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

(B) Preparation of calibration curve of molecular weight In the column, eluent, flow velocity, and detection wavelength described in the gel filtration conditions for HPLC analysis, a peptide having a known molecular weight dissolved in ultrapure water at 0.1 to 5 mg / mL as appropriate. 50 μL was injected and HPLC analysis was performed to confirm the retention time (Table 5). A calibration curve (Fig. 4) was prepared from the retention time and molecular weight.

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

(5) Calculation of Peptide Ratio of Molecular Weight of 800 to 1500 Da The peptide ratio of molecular weight of 800 to 1500 Da was calculated by the following formula.

The peptide ratios of the taste products (sample numbers 11 to 16) were as shown in Table 6.

(6) Analysis of carbohydrate concentration The carbohydrate concentration of the sample brewed product (Sample Nos. 11 to 16) based on the nutrition labeling standard was measured as described above. The results were as shown in Table 6.

(7) Sensory evaluation Each sample beverage was subjected to sensory evaluation by four trained panelists. Specifically, the two items of "taste thickness" and "presence or absence of miscellaneous taste" were evaluated according to the following criteria.

The "taste thickness" was evaluated on a 9-point scale of 1 to 9, and the average value of the evaluation scores of the four 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 No. 14 was evaluated as 3 points (no taste thickness), and 4.1 points or more were judged to have achieved a good flavor with respect to the taste thickness.

"Presence or absence of miscellaneous taste" was evaluated on a 9-point scale of 1 to 9, and the average value of the evaluation scores of the four panelists was calculated. Here, the "miscellaneous taste" means astringent taste, astringent taste, and astringent taste. Regarding "presence or absence of miscellaneous taste", 5 points were evaluated when there was miscellaneous taste, and 9 points were evaluated when there was very strong miscellaneous taste. A good flavor was achieved with 4.5 points or less. It was judged that it was done.

Based on the sensory evaluation results for "thickness of taste" and "presence or absence of miscellaneous taste", comprehensive evaluation was performed according to the following criteria.
A: The thickness of the taste is improved (5 points or more), and the taste is less (4.5 points or less).
B: The thickness of the taste is slightly improved (4.1 points or more), and the taste is less (4.5 points or less).
C: Although the thickness of the taste is improved (5 points or more), there is a miscellaneous taste (more than 4.5 points)
D: Although the thickness of the taste is slightly improved (4.1 points or more), there is a miscellaneous taste (4.5 points or more).
E: Taste thickness is not improved (less than 4.1 points)

(8) Evaluation results The results of the sensory evaluation are as shown in Table 6.

From the results in Table 6, even for beer-taste fermented alcoholic beverages with a sugar concentration of less than 1.1 g / 100 mL, beer with a molecular weight of 800 to 1500 Da and a peptide ratio of 25% or more realized a thick taste. It was confirmed that a taste fermented alcoholic beverage could be provided. In particular, it was confirmed that when the peptide ratio of the molecular weight of 800 to 1500 Da is 28 to 38%, it is possible to provide a beer-taste fermented alcoholic beverage in which the thickness of the taste is realized while suppressing the unpleasant taste.

Claims (7)

And a malt use ratio of 50% or more, and a low-alcohol beer carbohydrate concentration is less than 1.1 g / 100 mL, mass of all peptides derived from the beverage was fractionated by gel filtration for HPLC analysis the peptides of molecular weight of the beverage for 800～1500Da is the ratio of the peptide mass (HPLC analytical gel filtration method) (percentage) is 25% to 45%, molecular weight 800～1500Da (HPLC analytical gel filtration method) , A beer-taste beverage that is a peptide derived from a beer-taste fermented alcoholic beverage with a malt usage ratio of 50% or more . The beer-taste beverage according to claim 1, wherein the ratio of the mass of the peptide is 28 to 38%. The beer-taste beverage according to claim 1 or 2, which is a fermented alcoholic beverage. The beer-taste beverage according to any one of claims 1 to 3, wherein ungerminated wheat is used as at least a part of the raw material. A method for producing a beer-taste beverage having an improved flavor, a malt usage ratio of 50% or more, and a sugar concentration of less than 1.1 g / 100 mL, which is fractionated by a gel filtration method for HPLC analysis. Ri Na include adjusting the molecular weight 800~1500Da of the beverage to the mass of all peptides derived from the beverage the ratio of the mass of peptide (HPLC analytical gel filtration method) (percentage) in 25% to 45% was, A production method, wherein the peptide having a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) is a peptide derived from a beer-taste fermented alcoholic beverage having a malt usage ratio of 50% or more . A beer-taste fermented alcoholic beverage having a malt usage ratio of 50% or more contains a peptide having a molecular weight of 800 to 1500 Da (gel filtration method for HPLC analysis) as an active ingredient, and the malt usage ratio is 50% or more. , A flavor improver for beer-taste beverages having a sugar concentration of less than 1.1 g / 100 mL. A method for improving the flavor of a beer-taste beverage having a malt usage ratio of 50% or more and a sugar concentration of less than 1.1 g / 100 mL, which is derived from the beverage fractionated by a gel filtration method for HPLC analysis. Ri Na include adjusting the ratio of the mass of peptides of molecular weight 800～1500Da of the beverage to the mass of the whole peptide (HPLC analytical gel filtration) and (percentage) in 25% to 45%, molecular weight 800～1500Da ( A method for improving flavor, wherein the peptide of the gel filtration method for HPLC analysis) is a peptide derived from a beer-taste fermented alcoholic beverage having a malt use ratio of 50% or more .

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