JP7750643B2 - Beer-flavored beverages - Google Patents

Description

The present invention relates to a beer-taste beverage, a method for reducing the bitterness of a beer-taste beverage, and an agent for reducing the bitterness of a beer-taste beverage.

With the recent diversification of consumer tastes, there is a demand for the development of beer-flavored beverages with a variety of flavor and taste characteristics.

Hops or hop extracts are sometimes added to beer-flavored beverages to impart a moderate bitterness and aroma, but iso-α acids, the bitter components contained in hops, are known to have a bitter taste (afterbitterness) that tends to linger in the mouth, which can pose a challenge in product design. To address this issue, attempts have been made to mask the afterbitterness of iso-α acids by adding neotame as a masking agent (Patent Document 1).

JP 2012-244971 A

However, the neotame used in Patent Document 1 itself has a strong sweetness, which can affect the taste and aroma of the beverage, limiting product design. Furthermore, increasing the amount of specific flavor components to suppress the sharp bitterness can change the quality of the designed flavor, resulting in an unbalanced flavor.

The present invention relates to a method for reducing the sharp bitterness of a beer-taste beverage without disrupting the flavor balance, and to providing a beer-taste beverage with a reduced sharp bitterness. It also relates to providing a bitterness reducer for a beer-taste beverage that reduces the sharp bitterness without disrupting the flavor balance of the beer-taste beverage. Here, "sharp bitterness" refers to a bitterness with a narrow peak width that is felt immediately after drinking.

The present invention relates to the following [1] to [7].
[1] A beer-flavored beverage having an iso-α acid content of 10 to 30 ppm, a protein content of 35 to 50 kDa molecular weight of 20 ppm or more, and a ratio of the protein content of 35 to 50 kDa molecular weight to the iso-α acid content of 1.0 or more.
[2] A method for reducing the bitterness of a beer-flavored beverage having an iso-α acid content of 10 to 30 ppm, comprising adjusting the content of proteins having a molecular weight of 35 to 50 kDa in the beer-flavored beverage to 20 ppm or more and adjusting the ratio of the content of proteins having a molecular weight of 35 to 50 kDa to the content of iso-α acids to 1.0 or more.
[3] A beer-flavored beverage having an iso-α acid content of 30 to 50 ppm, a protein content of 35 to 50 kDa molecular weight of 20 ppm or more, and a ratio of the protein content of 35 to 50 kDa molecular weight to the iso-α acid content of 0.5 or more.
[4] A method for reducing the bitterness of a beer-flavored beverage having an iso-α acid content of 30 to 50 ppm, comprising adjusting the content of proteins having a molecular weight of 35 to 50 kDa in the beer-flavored beverage to 20 ppm or more and adjusting the ratio of the content of proteins having a molecular weight of 35 to 50 kDa to the content of iso-α acids to 0.5 or more.
[5] A beer-flavored beverage having an iso-α acid content of 50 to 150 ppm, a protein content of 35 to 50 kDa molecular weight of 30 ppm or more, and a ratio of the protein content of 35 to 50 kDa molecular weight to the iso-α acid content of 0.4 or more.
[6] A method for reducing the bitterness of a beer-flavored beverage having an iso-α acid content of 50 to 150 ppm, comprising adjusting the content of proteins having a molecular weight of 35 to 50 kDa in the beer-flavored beverage to 30 ppm or more and adjusting the ratio of the content of proteins having a molecular weight of 35 to 50 kDa to the content of iso-α acids to 0.4 or more.
[7] A bitterness mitigating agent for beer-flavored beverages, comprising a protein having a molecular weight of 35 to 50 kDa.

The present invention provides a method for reducing the sharp bitterness of a beer-taste beverage without disrupting the flavor balance, and a beer-taste beverage with a reduced sharp bitterness. It also provides a bitterness reducer for a beer-taste beverage that reduces the sharp bitterness without disrupting the flavor balance of the beer-taste beverage.

The inventors investigated the above-mentioned problem and surprisingly discovered that a protein with a molecular weight of 35 to 50 kDa has the effect of reducing the bitterness of beer-flavored beverages containing a specific amount of iso-α acids. The mechanism behind this is unclear, but because the protein with a molecular weight of 35 to 50 kDa itself has no flavor, it is possible to reduce the bitterness without disrupting the flavor balance of the beer-flavored beverage.

The beer-flavored beverage of the present invention contains iso-α acids and a protein with a molecular weight of 35 to 50 kDa.

The iso-α acid content in the beer-taste beverage of the present invention can be 10 to 150 ppm. In this specification, the iso-α acid content is measured using a high-performance liquid chromatograph (HPLC) with a UV detector, with reference to the content described in J. Am. Soc. Brew. Chem., 43:136 (1985).

The content of proteins with a molecular weight of 35 to 50 kDa and the ratio of the content of proteins with a molecular weight of 35 to 50 kDa to the content of iso-α acids (proteins with a molecular weight of 35 to 50 kDa/iso-α acids) in the beer-taste beverage of the present invention will vary depending on the content of iso-α acids.

When the iso-α acid content is 10-30 ppm, the content of proteins with a molecular weight of 35-50 kDa is, from the viewpoint of reducing bitterness, 20 ppm or more, preferably 25 ppm or more, more preferably 35 ppm or more, and even more preferably 45 ppm or more. From the viewpoint of flavor balance, it is preferably 300 ppm or less, more preferably 170 ppm or less, and even more preferably 80 ppm or less, or any combination of these ranges is acceptable. Furthermore, from the viewpoint of reducing bitterness, the ratio of the content of proteins with a molecular weight of 35-50 kDa to the content of iso-α acids is, from the viewpoint of reducing bitterness, 1.0 or more, preferably 1.3 or more, more preferably 1.8 or more, and even more preferably 2.3 or more. From the viewpoint of flavor balance, it is preferably 15.0 or less, more preferably 8.0 or less, and even more preferably 4.0 or less, or any combination of these ranges is acceptable.

When the iso-α acid content is 30-50 ppm, the content of proteins with a molecular weight of 35-50 kDa is 20 ppm or more, preferably 25 ppm or more, and more preferably 35 ppm or more, from the viewpoint of reducing bitterness; and from the viewpoint of flavor balance, it is preferably 300 ppm or less, more preferably 170 ppm or less, and even more preferably 80 ppm or less, or any combination of these ranges is acceptable. Furthermore, the ratio of the content of proteins with a molecular weight of 35-50 kDa to the content of iso-α acids is 0.5 or more, preferably 0.7 or more, and more preferably 0.9 or more, from the viewpoint of reducing bitterness; and from the viewpoint of flavor balance, it is preferably 7.5 or less, more preferably 4.2 or less, and even more preferably 2.0 or less, or any combination of these ranges is acceptable.

When the iso-α acid content is 50 to 150 ppm, preferably 50 to 100 ppm, more preferably 60 to 100 ppm, and even more preferably 70 to 90 ppm, the content of proteins with a molecular weight of 35 to 50 kDa is, from the viewpoint of reducing bitterness, 30 ppm or more, preferably 35 ppm or more, more preferably 45 ppm or more, and even more preferably 65 ppm or more. From the viewpoint of flavor balance, it is preferably 300 ppm or less, more preferably 170 ppm or less, and even more preferably 120 ppm or less, or any combination of these ranges is acceptable. Furthermore, from the viewpoint of reducing bitterness, the ratio of the content of proteins with a molecular weight of 35 to 50 kDa to the content of iso-α acids is 0.4 or more, preferably 0.5 or more, and even more preferably 0.9 or more, from the viewpoint of reducing bitterness. From the viewpoint of flavor balance, it is preferably 3.5 or less, more preferably 2.0 or less, and even more preferably 1.5 or less, or any combination of these ranges is acceptable.

Methods for adjusting the mass ratio of proteins with a molecular weight of 35 to 50 kDa within the above range include adding proteins with a molecular weight of 35 to 50 kDa, using raw materials with a high content of 35 to 50 kDa proteins, and controlling the content of 35 to 50 kDa proteins through fermentation conditions.

When a protein with a molecular weight of 35 to 50 kDa is added as a bitterness mitigating agent, a preferred embodiment is to add a bitterness mitigating agent containing a barley-derived protein. Examples of such barley include barley, wheat, rye, oats, oats, and oats, with barley being preferred. Furthermore, either germinated or ungerminated barley may be used, with germinated barley malt being preferred. These may be added alone or in combination of two or more types.

When controlling the content of 35-50 kDa proteins through fermentation conditions, it is also possible to control this by controlling the fermentation temperature. For example, by using a lower temperature, the loss of 35-50 kDa proteins due to foam separation during fermentation can be reduced, resulting in more 35-50 kDa proteins being contained in the beer.

In this specification, quantification of 35-50 kDa proteins is performed using the Lowry method. The specific measurement method is shown below.

1. Purification of 35-50 kDa Proteins 1) Protein Concentration with Ammonium Sulfate 3,900 g of ammonium sulfate (60% saturated ammonium sulfate) was added to 10 L of beer, stirred for 3 hours, and then centrifuged at 12,000 g at 4°C for 1 hour to obtain a precipitate. The resulting precipitate was suspended in as little 20 mM phosphate buffer (pH 9.0) as possible, and 20 mM phosphate buffer (pH 9.0) was added until turbidity subsided. Ultrafiltration was then performed using an Amicon Ultra-15 (10 kDa cutoff, Merck, UFC901024) at 2,800 g at 4°C to a final volume of approximately 1 ml. Approximately 10 ml of 20 mM phosphate buffer (pH 9.0) was then added, and the mixture was centrifuged again under the same conditions to remove unnecessary ammonium sulfate. The resulting supernatant was used as the beer protein-enriched fraction for the next step.

2) Fractionation of beer protein enriched fraction using cationic resin
An Econo-column is packed with 100 ml of SP Sepharose Fast Flow (GE Healthcare Life Sciences), and equilibrated with 300 ml of water and then 300 ml of 20 mM acetate buffer (pH 4.5).
A beaker was prepared, and the beer protein enriched fraction obtained in step 1 was mixed with 20 mM acetate buffer (pH 4.5) to bring the volume to 400 ml. 100 ml of SP Sepharose equilibrated with acetate buffer (pH 4.5) was added to the resulting solution, and batch adsorption was carried out for 3 hours, stirring approximately every 10 minutes with a spatula. The contents of the beaker were then packed into a column, and the flow-through fraction was collected (FT). Next, 300 ml of 20 mM acetate buffer (pH 4.5) was loaded (A). The following loads were then added: 300 ml of 20 mM acetate buffer (pH 4.5) containing 0.1 M NaCl (B), 300 ml of 20 mM acetate buffer (pH 4.5) containing 0.2 M NaCl (C), 300 ml of 20 mM acetate buffer (pH 4.5) containing 0.3 M NaCl (D), and 300 ml of 20 mM acetate buffer (pH 4.5) containing 0.5 M NaCl (E). The obtained fractions were analyzed by SDS-PAGE, and fractions containing 35-50 kDa proteins were collected. These fractions containing 35-50 kDa proteins were used in the next step as the beer protein cation exchange resin-bound fraction.

3) Concentration of the beer protein cation exchange resin-bound fraction with ammonium sulfate: Add 430 g of ammonium sulfate to 1 L of the beer protein cation exchange resin-bound fraction in a beaker while stirring. After stirring for 3 hours, transfer the contents of the beaker to a centrifuge tube and centrifuge (12,000 g, 4°C, 3 hours) to obtain a precipitate.
The precipitate was suspended in the smallest possible amount of 20 mM acetate buffer (pH 4.5), and 20 mM acetate buffer (pH 4.5) was added until the turbidity disappeared. Ultrafiltration was then performed using an Amicon Ultra-15 (10 kDa cutoff, Merck, UFC901024) at 2,800 g and 4°C to a final volume of approximately 1 ml. Approximately 10 ml of 20 mM acetate buffer (pH 4.5) was then added, and the mixture was centrifuged again under the same conditions to remove any unnecessary ammonium sulfate. The resulting supernatant was used as the beer protein cation exchange resin-bound fraction concentrate for the next step.

4) Fractionation of beer protein cation exchange resin-bound fraction concentrate using an anion exchange resin
100 ml of Q Sepharose Fast Flow (GE Healthcare Life Sciences) was packed into an Econo-column, and 300 ml of water was passed through it, followed by 300 ml of 20 mM phosphate buffer (pH 9.0) to equilibrate it.
Prepare a beaker and add 20 mM phosphate buffer (pH 9.0) to the beer protein cation exchange resin-bound fraction concentrate obtained in step 3 to bring the volume to 400 ml. Add 100 ml of Q Sepharose equilibrated with 20 mM phosphate buffer (pH 9.0) to the resulting solution, and allow batch adsorption to occur over 3 hours, stirring with a spatula approximately every 10 minutes. The contents of the beaker are then packed into a column, and the flow-through fraction is collected (FT). Next, load 500 ml of 20 mM phosphate buffer (pH 9.0) (A). The following fractions were loaded: 300 ml of 20 mM phosphate buffer (pH 9.0) containing 0.1 M NaCl (B), 300 ml of 20 mM phosphate buffer (pH 9.0) containing 0.2 M NaCl (C), 300 ml of 20 mM phosphate buffer (pH 9.0) containing 0.3 M NaCl (D), and 300 ml of 20 mM phosphate buffer (pH 9.0) containing 0.5 M NaCl (E). The resulting FT and fractions A through E were analyzed by SDS-PAGE, and fractions containing 35-50 kDa proteins were collected. These fractions containing 35-50 kDa proteins were used in the next step as the 35-50 kDa protein ion-exchange resin-bound fraction.
For (A), (B), (C), (D), and (E), immediately after column elution, neutralize by adding 15 ml of 0.5 M disodium phosphate to 500 ml of solution. Step 4) should be performed within one day to minimize changes in the protein due to alkali.

5) Concentration of the 35-50 kDa protein ion exchange resin-bound fraction with ammonium sulfate: Add 430 g of ammonium sulfate to 1 L of the 35-50 kDa protein ion exchange resin-bound fraction in a beaker while stirring. After stirring for 3 hours, transfer the contents of the beaker to a centrifuge tube and centrifuge (12,000 g, 4°C, 3 hours) to obtain a precipitate.
The precipitate was suspended in the smallest possible amount of 20 mM acetate buffer (pH 4.5), and 20 mM acetate buffer (pH 4.5) was added until the turbidity disappeared. Ultrafiltration was then performed using an Amicon Ultra-15 (10 kDa cutoff, Merck, UFC901024) (centrifugation at 2,800 g at 4°C to a final volume of approximately 1 ml). Approximately 10 ml of 20 mM acetate buffer (pH 4.5) was then added, and the mixture was centrifuged again under the same conditions to remove any unnecessary ammonium sulfate. The resulting supernatant was used for analysis as the 35-50 kDa purified protein.

2. Protein Quantification by the Lowry Method Concentration occurs during the preparation of a 35-50 kDa protein purified product. Specifically, the volume of a 35-50 kDa protein purified product obtained from a beer-taste beverage is smaller than the volume of the beer-taste beverage. Therefore, the protein content of a 35-50 kDa protein purified product from a beer-taste beverage of the present invention is calculated by dividing the protein content measured using a 35-50 kDa protein purified product prepared from a beer-taste beverage by the concentration factor (i.e., the ratio obtained by dividing the volume of the beer-taste beverage used in the preparation by the volume of the 35-50 kDa protein purified product obtained in the preparation) when the 35-50 kDa protein purified product was prepared from the beer-taste beverage. Protein quantification was performed by the Lowry method using a commercially available kit (DC Protein Assay, Bio-Rad). First, the concentration of the fraction was adjusted to an appropriate range. To 5 μL of the adjusted concentration sample, 50 μL of Solution A was added and stirred, followed by 400 μL of Solution B, which was then added and stirred. After a 15-minute color reaction at room temperature, 350 μL was transferred to a 96-well plate and the absorbance at 750 nm was measured. The peptide concentration (mg/mL) was calculated based on the absorbance obtained and a previously prepared calibration curve. The calibration curve was prepared using BSA (bovine serum albumin). The content of 35-50 kDa protein in the 35-50 kDa protein purified product was calculated based on this calibration curve.

The method for producing a beer-taste beverage of the present invention is not particularly limited, but an example is a production method that includes a step of adding a 35-50 kDa protein. More specifically, an embodiment in which a barley-derived protein with a molecular weight of 35-50 kDa is added as a bitterness mitigator is exemplified below. The bitterness mitigator may optionally contain any known additive that can be added to beverages, as long as it does not impair the effects of the present invention. In another embodiment, the bitterness mitigator may be increased by changing the production process conditions to produce more barley-derived protein with a molecular weight of 35-50 kDa than conventional conditions, such as by using a high-nitrogen malt raw material from North America that has a high content of 35-50 kDa protein, or by increasing the amount of 35-50 kDa protein through fermentation conditions.

The manufacturing method of this embodiment in which a barley-derived protein with a molecular weight of 35 to 50 kDa is added as a bitterness mitigator is the same as the manufacturing method of a general beer-taste beverage, except for the step of adding the 35 to 50 kDa protein. Examples of manufacturing methods for beer-taste beverages are provided below. Beer-taste beverages can be manufactured using either malt or no malt as an ingredient, and can be manufactured as follows:

Beer-flavored beverages containing alcohol produced using malt as a raw material are first prepared by adding enzymes such as amylase to a mixture containing malt and other barley, as well as other grains, starch, sugars, bittering agents, or coloring agents, as needed, and water, followed by gelatinization and saccharification, followed by filtration to produce a saccharified liquid. Hops and bittering agents, as needed, are added to the saccharified liquid, which is then boiled and solids such as coagulated proteins are removed in a clarifying tank. As an alternative to this saccharified liquid, hops may be added to malt extract and warm water, and then boiled. Hops may be added at any stage, from the start of boiling to the end of boiling. Known conditions may be used for the saccharification, boiling, and solids removal processes. Known conditions may be used for the fermentation and storage processes. The resulting fermented liquid is filtered, and carbon dioxide gas is added to the filtrate. The liquid is then filled into containers and sterilized to produce the desired beer-flavored beverage. Grain-derived spirits may also be added as an alcoholic component. Spirits refer to alcoholic beverages obtained by fermenting grains such as barley, rice, buckwheat, and corn using yeast, followed by distillation. Barley is the preferred grain used to make spirits. In each of the above processes, the 35-50 kDa protein may be added at any stage up to the filling stage.

Beer-flavored beverages containing alcohol that are produced without using malt as a raw material are prepared by mixing liquid sugar containing a carbon source, a nitrogen source (barley or a non-malt amino acid-containing material), hops, coloring, and other ingredients with warm water to form a liquid sugar solution. This liquid sugar solution is then boiled. When hops are used as an ingredient, the hops may be added to the liquid sugar solution during boiling rather than before the start of boiling. As an alternative to this saccharified solution, hops may be added to an extract made from ingredients other than malt, to which warm water is added, and the mixture is then boiled. The hops may be added at any stage from the start of boiling to the end of boiling. Known conditions may be used for the fermentation and storage processes. The resulting fermented liquid is filtered, and carbon dioxide gas is added to the filtrate. The resulting beverage is then filled into containers and sterilized to obtain the desired beer-flavored beverage. Grain-derived spirits may also be added as an alcoholic component. Spirits refer to alcoholic beverages obtained by fermenting grains such as barley, rice, buckwheat, or corn using yeast, followed by distillation. Barley is the preferred grain used as a raw material for spirits. In each of the above steps, the 35-50 kDa protein may be added at any stage up to the filling stage.

Beer-flavored beverages that are non-fermented and contain alcohol may or may not use malt, and may be those in which the alcohol content of the final product is adjusted by adding raw material alcohol or the like. The raw material alcohol may be added at any stage from the saccharification process to the filling process. Grain-derived spirits may also be added as an alcohol component. Spirits refer to alcoholic beverages obtained by fermenting grains such as barley, rice, buckwheat, and corn using yeast, followed by distillation. Barley is preferred as the grain used as the raw material for spirits. In each of the above processes, the step of adding the 35-50 kDa protein may be performed at any stage up to the filling process.

Non-alcoholic beer-flavored beverages produced using malt as a raw material are first prepared by adding enzymes such as amylase to a mixture containing malt and other barley, as well as other grains, starch, sugars, bittering agents, or coloring agents, as needed, and water, followed by gelatinization and saccharification, filtration, and production of a saccharified liquid. Hops and bittering agents, as needed, are added to the saccharified liquid, which is then boiled and solids such as coagulated proteins are removed in a clarifying tank. As an alternative to this saccharified liquid, hops may be added to malt extract and warm water, and then boiled. Hops may be added at any stage, from the start of boiling to before the end of boiling. Known conditions may be used for the saccharification, boiling, and solids removal processes. After boiling, the resulting wort is filtered, and carbon dioxide gas is added to the filtrate. The resulting liquid is then filled into containers and sterilized to produce the desired non-alcoholic beer-flavored beverage. In each of the above processes, the addition of the 35-50 kDa protein may be performed at any stage up to the filling stage.

When producing a non-alcoholic beer-flavored beverage that does not use malt as an ingredient, first, liquid sugar containing a carbon source, a nitrogen source as an amino acid-containing material other than barley or malt, hops, coloring, etc. are mixed with warm water to form a liquid sugar solution. This liquid sugar solution is then boiled. If hops are used as an ingredient, the hops may be mixed into the liquid sugar solution during boiling rather than before the start of boiling. Carbon dioxide gas is added to the boiled liquid sugar solution. The resulting solution is then filled into containers and sterilized to obtain the desired non-alcoholic beer-flavored beverage. In each of the above steps, the 35-50 kDa protein may be added at any step up to filling.

In the production method of this embodiment, an aliphatic alcohol may be added to impart a boozy flavor. There are no particular limitations on the aliphatic alcohol as long as it is a known alcohol, but aliphatic alcohols with 4 to 5 carbon atoms are preferred. In the production method of this embodiment, preferred aliphatic alcohols include those with 4 carbon atoms such as 2-methyl-1-propanol and 1-butanol, and those with 5 carbon atoms such as 3-methyl-1-butanol, 1-pentanol, and 2-pentanol. These may be used alone or in combination of two or more. The content of the aliphatic alcohol with 4 to 5 carbon atoms is preferably 0.0002 to 0.0007% by mass, more preferably 0.0003 to 0.0006% by mass. In this specification, the content of the aliphatic alcohol can be measured using headspace gas chromatography.

(acidulant)
The acidulant used in the production method of this embodiment is preferably one or more acids selected from the group consisting of citric acid, lactic acid, phosphoric acid, and malic acid. In addition, in the production method of this embodiment, acids other than the above acids, such as succinic acid, tartaric acid, fumaric acid, and glacial acetic acid, can also be used. These acids can be used without limitation as long as they are approved for addition to foods. In the production method of this embodiment, it is preferable to use a combination of lactic acid, which appropriately imparts a mellow sourness, and phosphoric acid, which appropriately imparts a slightly pungent sourness.

The content of the acidulant in the beer-taste beverage obtained by the production method of this embodiment, calculated as citric acid, is preferably 200 ppm or more, more preferably 550 ppm or more, and even more preferably 700 ppm or more, from the viewpoint of imparting a beer-taste. From the viewpoint of sourness, the content is preferably 15,000 ppm or less, more preferably 5,500 ppm or less, and even more preferably 2,000 ppm or less. Therefore, in this embodiment, the content of the acidulant, calculated as citric acid, is preferably in the range of 200 ppm to 15,000 ppm, preferably 550 ppm to 5,500 ppm, and more preferably 700 ppm to 1,500 ppm. In this specification, the citric acid equivalent amount refers to the amount calculated from the acidity of each acidulant using the acidity of citric acid as the standard. For example, the citric acid equivalent amount equivalent to 100 ppm of lactic acid is 120 ppm, the citric acid equivalent amount equivalent to 100 ppm of phosphoric acid is 200 ppm, and the citric acid equivalent amount equivalent to 100 ppm of malic acid is 125 ppm.

The amount of acidulant contained in beer-flavored beverages is calculated by analysis using high-performance liquid chromatography (HPLC) or other methods.

(hop)
In the production method of this embodiment, hops can be used as part of the raw materials. Since the flavor tends to be similar to that of beer, it is desirable to use hops as part of the raw materials. When using hops, ordinary pelleted hops, powdered hops, and hop extracts used in the production of beer and the like can be appropriately selected and used depending on the desired flavor. In addition, processed hop products such as isomerized hops and reduced hops may also be used. These products are included in the hops used in the production method of this embodiment. Furthermore, the amount of hops added is not particularly limited, but is typically about 0.0001 to 1% by mass of the total amount of the beverage.

(Other ingredients)
In the production method of this embodiment, other ingredients may be used as desired, such as sweeteners (including high-intensity sweeteners), bittering agents, flavorings, yeast extracts, coloring agents such as caramel color, plant-extracted saponin substances such as soybean saponin and quillaja saponin, plant protein and peptide-containing substances such as corn and soybean, animal protein such as whey, seasonings such as dietary fiber and amino acids, and antioxidants such as ascorbic acid, as long as they do not impair the effects of this embodiment.

From the perspective of improving the flavor of the beverage, the pH of the beer-taste beverage obtained by this manufacturing method is preferably 3.0 to 5.0, more preferably 3.0 to 4.5, and even more preferably 3.0 to 4.0.

(packaged beverages)
The beer-taste beverage obtained by the production method of this embodiment can be packaged in containers. The type of container is not particularly limited, and the beverage can be filled into a sealed container such as a bottle, can, barrel, or plastic bottle to produce a packaged beverage.

The present invention will be explained in more detail below using examples, but the present invention is not limited to the following examples.

Preparation Example 1: Preparation of 35-50 kDa Proteins 35-50 kDa proteins were prepared according to the procedure described above in the section "Purification of 35-50 kDa proteins."

Preparation Example 2: Preparation of amino acid mixture
Various amino acid reagents were mixed to prepare an amino acid mixture so that the amino acid ratio was the same as that of the amino acid sequence of Protein Z shown in Fig. 4 in Eur Food Res Technol (2010) 230:665-673.

Preparation of beer-taste beverages Reference Examples 1 to 3, Examples 1 to 26, Comparative Example 7
30 kg of malt was crushed to an appropriate particle size and placed in a mashing tank. 120 L of warm water was added to produce a mash at approximately 50°C. A portion of the mash was heated to 100°C and boiled, while the remainder was subjected to saccharification. After saccharification was completed, the mash was heated to 78°C and transferred to a wort filtration tank, where it was filtered to obtain a filtrate. Brewer's yeast was added to the filtrate, and fermentation was carried out at approximately 15°C for approximately 15 days to obtain a stored beer. The stored beer was then filtered to remove the yeast without heat treatment, and a beer-taste beverage for evaluation was produced. The resulting beer-taste beverage was analyzed according to the above-mentioned measurement method, and the content of 35-50 kDa proteins was found to be 12 ppm. Iso-α acids (ISOHOP (John I. HAAS)) and the 35-50 kDa protein of Preparation Example 1 were added to this beer-taste beverage to the concentrations shown in Tables 1, 3, and 5, to obtain the beer-taste beverages of Reference Examples 1-3, Examples 1-26, and Comparative Example 7.

Comparative Examples 1 to 6, 8 to 10
Beer-taste beverages were obtained in the same manner as in Examples 1 to 26, except that the 35-50 kDa protein of Preparation Example 1 was replaced with the amino acid mixture of Preparation Example 2 at the concentrations shown in Tables 2, 4, and 6.

Flavor Evaluation The flavors of Examples 1 to 26 and Comparative Examples 1 to 10 were evaluated by a sensory test. Five well-trained sensory evaluators evaluated the flavors on a scale of 1 to 5 for "intensity of sharp bitterness" and "overall flavor balance."

For the sharp bitterness suppression effect, "very noticeable" was assigned 5 points, "slightly noticeable" was assigned 4 points, "slightly noticeable" was assigned 3 points, "slightly noticeable" was assigned 2 points, and "not noticeable" was assigned 1 point. An average score was calculated and evaluation was conducted according to the following criteria. For the overall flavor balance, "very good" was assigned 5 points, "good" was assigned 4 points, "slightly good" was assigned 3 points, "slightly bad" was assigned 2 points, and "bad" was assigned 1 point. An average score was calculated and evaluation was conducted according to the following criteria. The results are shown in Tables 1 to 6. For all sensory evaluations, Reference Example 1 was assigned 1 point for Tables 1 and 2, Reference Example 2 was assigned 1 point for Tables 3 and 4, and Reference Example 3 was assigned 1 point for Tables 5 and 6.

<Evaluation criteria>
×: average value 1.0 or more but less than 2.0 △: average value 2.0 or more but less than 3.0 ○: average value 3.0 or more but less than 4.0 ◎: average value 4.0 or more but less than 5.0

As can be seen from Tables 1 to 6, the beer-taste beverages of Examples 1 to 18, which contained 20 to 40 ppm iso-α acids and 20 to 200 ppm 35 to 50 kDa proteins, and the beer-taste beverages of Examples 19 to 26, which contained 80 ppm iso-α acids and 30 to 200 ppm 35 to 50 kDa proteins, had a reduced sharp bitterness and excellent flavor balance. On the other hand, in Comparative Examples 1 to 6 and 8 to 10, which contained the addition of an amino acid mixture, although the bitterness was reduced, there were problems with the flavor balance.

The present invention makes it possible to provide a beer-flavored beverage with a reduced sharp bitterness.

Claims (7)

A beer-flavored beverage having an iso-α acid content of 10 to 30 ppm, a protein content of 35,000 to 50,000 molecular weight of 20 ppm or more, a ratio of the protein content of 35,000 to 50,000 molecular weight to the iso-α acid content of 2.3 to 15, and the protein of molecular weight 35,000 to 50,000 is a protein derived from a beer-flavored beverage containing barley as an ingredient. A method for reducing the bitterness of a beer-taste beverage containing 10 to 30 ppm of iso-α acids, comprising a step of adjusting the content of proteins with a molecular weight of 35,000 to 50,000 in the beer-taste beverage so that the content is 20 ppm or more and the ratio of the content of proteins with a molecular weight of 35,000 to 50,000 to the content of iso-α acids is 1.0 or more, wherein the proteins with a molecular weight of 35,000 to 50,000 are proteins derived from a beer-taste beverage containing barley as an ingredient. A beer-flavored beverage having an iso-α acid content of 30 to 50 ppm, a protein content of 35,000 to 50,000 molecular weight of 20 ppm or more, a ratio of the protein content of 35,000 to 50,000 molecular weight to the iso-α acid content of 0.9 to 7.5, and the protein of molecular weight 35,000 to 50,000 being a protein derived from a beer-flavored beverage containing barley as an ingredient. A method for reducing the bitterness of a beer-taste beverage containing 30 to 50 ppm of iso-α acids, comprising a step of adjusting the content of proteins with a molecular weight of 35,000 to 50,000 in the beer-taste beverage to 20 ppm or more and the ratio of the content of proteins with a molecular weight of 35,000 to 50,000 to the content of iso-α acids to 0.5 or more, wherein the proteins with a molecular weight of 35,000 to 50,000 are proteins derived from a beer-taste beverage containing barley as an ingredient. A beer-flavored beverage having an iso-α acid content of 50 to 150 ppm, a protein content of 35,000 to 50,000 molecular weight of 30 ppm or more, a ratio of the protein content of 35,000 to 50,000 molecular weight to the iso-α acid content of 0.9 to 3.5, and the protein of molecular weight 35,000 to 50,000 being a protein derived from a beer-flavored beverage containing barley as an ingredient. A method for reducing the bitterness of a beer-taste beverage containing 50 to 150 ppm of iso-α acids, comprising a step of adjusting the content of proteins with a molecular weight of 35,000 to 50,000 in the beer-taste beverage to 30 ppm or more and the ratio of the content of proteins with a molecular weight of 35,000 to 50,000 to the content of iso-α acids to 0.4 or more, wherein the proteins with a molecular weight of 35,000 to 50,000 are proteins derived from a beer-taste beverage containing barley as an ingredient. The bitterness mitigating agent for beer-flavored beverages contains iso-α acid, which is a protein having a molecular weight of 35,000 to 50,000, the protein being derived from a beer-flavored beverage containing barley as a raw material.