Beer-flavored nonalcoholic beverage

Abstract

The purpose of the present invention is to provide a novel beer-flavored nonalcoholic beverage in which an overly sweet taste or aroma is suppressed, and a beer-like balance is achieved. The present invention provides a beer-flavored nonalcoholic beverage in which malt is used as at least a portion of the starting materials. The concentration of peptides in the beverage is 0.25 to 3.00 mg / mL, the peptides having a molecular weight of 800 to 4600 Da as fractionated by gel filtration for HPLC (gel filtration for HPLC).

Description

Beer-flavored non-alcoholic beverage

[0001] The present invention relates to a beer-flavored non-alcoholic beverage.

[0002] In addition to the recent rise in health consciousness, the demand for beer-flavored non-alcoholic beverages has been increasing year by year due to the diversification of drinking occasions and consumer preferences. In the field of beer-flavored non-alcoholic beverages, various technologies have been developed to enable non-alcoholic beverages to have the distinctive taste and aroma of beer (Patent Documents 1 and 2). However, in light of the increasing level of consumer preferences, it can be said that there is still a need for improved flavor in beer-flavored non-alcoholic beverages.

[0003] Japanese Patent No. 6786699 Japanese Patent Application Laid-Open No. 2019-154317

[0004] The present inventors have discovered a problem in that beer-flavored non-alcoholic beverages tend to have a stronger sweet flavor than regular beer due to the residual sugars that tend to remain during production. The present invention aims to provide a novel beer-flavored non-alcoholic beverage that has a reduced sweet flavor and aroma and achieves a beer-like balance.

[0005] The present inventors have found that by incorporating a peptide with a molecular weight of 800 to 4600 Da into a non-alcoholic beer-flavored beverage at a concentration within a predetermined range, the overly sweet taste and aroma of the beverage can be suppressed while still achieving a beer-like balance. The present invention is based on this finding.

[0006] The present invention provides the following: [1] A beer-flavored non-alcoholic beverage made at least in part from malt, wherein the concentration of peptides with molecular weights of 800 to 4,600 Da (as determined by gel filtration for HPLC analysis) in the beverage fractionated by gel filtration for HPLC analysis is 0.25 mg / mL or more and 3.00 mg / mL or less. [2] The beer-flavored non-alcoholic beverage according to [1] above, wherein the concentration of a genuine extract in the beverage is 0.3 w / w% or more and 10 w / w% or less. [3] The beer-flavored non-alcoholic beverage according to [1] or [2] above, wherein the peptide concentration is 1.00 mg / mL or more and 3.00 mg / mL or less. [4] The beer-flavored non-alcoholic beverage according to any one of [1] to [3] above, wherein the malt usage ratio is 25% by mass or more. [5] The beer-flavored non-alcoholic beverage according to any one of [1] to [4], wherein the beer-flavored non-alcoholic beverage is a dealcoholic beverage. [6] A method for producing a beer-taste non-alcoholic beverage using malt as at least a part of its raw material, comprising a step of incorporating peptides having a molecular weight of 800 to 4,600 Da (gel filtration for HPLC analysis) into the beverage so that the concentration of the peptides in the beverage fractionated by gel filtration for HPLC analysis is 0.25 mg / mL or more and 3.00 mg / mL or less. [7] The production method according to [6] above, comprising a step of removing alcohol from a fermented wort broth. [8] A method for improving the flavor of a beer-taste non-alcoholic beverage using malt as at least a part of its raw material, comprising a step of incorporating peptides in the beverage fractionated by gel filtration for HPLC analysis so that the concentration of the peptides in the beverage fractionated by gel filtration for HPLC analysis is 0.25 mg / mL or more and 3.00 mg / mL or less.

[0007] According to the present invention, by incorporating peptides with molecular weights of 800 to 4600 Da into a beer-flavored non-alcoholic beverage so that the concentration of said peptides falls within a predetermined numerical range, it is possible to provide a beer-flavored non-alcoholic beverage in which the overly sweet taste and aroma are suppressed and a beer-like balance is achieved, which is advantageous in that it can meet the diverse preferences and needs of consumers.

[0008] FIG. 1 shows a calibration curve prepared from retention times and molecular weights of known substances in a gel filtration method for HPLC analysis carried out in Example 1 of the Examples. Specific Description of the Invention

[0009] In the present invention, the term "beer-flavored non-alcoholic beverage" refers to a beverage with a beer-like flavor, and is used to include both beverages that contain no alcohol at all, i.e., beverages with an ethanol concentration of 0.00 v / v%, and beverages with an ethanol concentration of more than 0.00 v / v% and less than 1 v / v%. Beer-flavored non-alcoholic beverages include fermented beer-flavored non-alcoholic beverages and non-fermented beer-flavored non-alcoholic beverages. Fermented beer-flavored non-alcoholic beverages refer to beer-flavored non-alcoholic beverages that have undergone a fermentation process using yeast, and include beverages produced by removing the alcoholic components generated during the fermentation process after the fermentation process (sometimes referred to as "de-alcoholized beverages" in this specification), as well as beverages produced by terminating the fermentation process when the ethanol concentration is less than 1 v / v%.

[0010] In the non-alcoholic beer-taste beverage of the present invention, the upper limit value (or less) of the alcohol (ethanol) concentration is 0.99% (v / v), 0.98% (v / v), 0.97% (v / v), 0.96% (v / v), 0.95% (v / v), 0.94% (v / v), 0.93% (v / v), 0.92% (v / v), 0.91% (v / v), 0.90% (v / v), 0.89% (v / v), 0.88% (v / v), 0.87% (v / v), 0.86% (v / v), 0.85% (v / v), 0.84% (v / v), 0.83% (v / v), 0.82% (v / v), 0.81% (v / v), 0.80% (v / v), 0.79% (v / v), 0.78% (v / v), 0.77% (v / v), 0.76% (v / v), 0.75% (v / v), 0.74% (v / v), 0.73% (v / v), 0.72% (v / v), 0.71% (v / v), 0.70% (v / v), 0.69% (v / v), 0.68% (v / v), 0.67% (v / v), 0.66% (v / v), 0.65% (v / v), 0.64% (v / v), 0.63% (v / v), 0.62% (v / v), 0.61% (v / v), 0.60% (v / v), 0.59% (v / v), 0.58% (v / v), 0.57% (v / v), 0.56% (v / v), 0.55% (v / v), 0.54% (v / v), 0.53% (v / v), 0.52% (v / v), 0.51% (v / v), 0.50% (v / v), 0.49% (v / v), 0.48% (v / v), 0.47% (v / v), 0.46% (v / v), 0.45% (v / v), 0.44% (v / v), 0.43% (v / v), 0.42% (v / v), 0.41% (v / v), 0.40% (v / v), 0.39% (v / v), 0.38% (v / v), 0.37% (v / v), 0.36% (v / v), 0.35% (v / v), 0.34% (v / v), 0.33% (v / v), 0.32% (v / v), 0.31% (v / v), 0.30% (v / v), 0.29% (v / v), 0.28% (v / v), 0.27% (v / v), 0.26% (v / v), 0.25% (v / v), 0.24% (v / v), 0.23% (v / v), 0.22% (v / v), 0.21% (v / v), 0.20% (v / v), 0.19% (v / v), 0.18% (v / v), 0.17% (v / v), 0.16% (v / v), 0.15% (v / v),0.14% (v / v), 0.13% (v / v), 0.12% (v / v), 0.11% (v / v), 0.10% (v / v), 0.09% (v / v), 0.08% (v / v), 0.07% (v / v), 0.0 6% (v / v), 0.05% (v / v), 0.04% (v / v), 0.03% (v / v), 0.02% (v / v), 0.01% (v / v), 0.009% (v / v), 0.008% (v / v), 0.00 The upper limit may be 0.001% (v / v), 0.002% (v / v), 0.003% (v / v), or 0.004% (v / v), and the lower limit (not less than or exceeding) may be 0.001% (v / v), 0.002% (v / v), 0.003% (v / v), or 0.004% (v / v). These upper and lower limits may also be combined in any manner.

[0011] The alcohol concentration (ethanol concentration) in a beverage can be measured by gas chromatography (GC) with an FID detector. To achieve more accurate concentration measurements, it is desirable to use a calibration curve created based on the measurements of several control samples with known concentrations. It is preferable that these control samples with known concentrations are in the same range as the concentration to be measured. It is also preferable to use an internal standard, such as 2-propanol.

[0012] The beer-taste non-alcoholic beverage of the present invention uses malt as at least a portion of its raw material, and preferably uses at least one or both of barley malt and wheat malt as the barley-derived raw material. In the present invention, the proportion of barley malt (by mass) in the raw material malt (by mass) can be, for example, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 100%, and the proportion of wheat malt (by mass) in the raw material malt (by mass) can be, for example, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 100%. It should be noted that the present invention can also use extracted malt (i.e., malt extract). In the above, "or more" can be interpreted as "exceeding."

[0013] The beer-flavored non-alcoholic beverage of the present invention can suppress sweet taste and aroma and achieve a beer-like balance regardless of the malt content ratio, and in this case, the malt content ratio can be, for example, 10% or less, 10% or more, 15% or less, 15% or more, 20% or less, 20% or more, 25% or more, 30% or less, 30% or more, 40% or less, 40% or more, 50% or less, 50% or more, 55% or less, 55% or more, 60% or less, 60% or more, 65% or less, 65% or more, 70% or less, 70% or more, 75% or less, 75% or more, 80% or less, 80% or more, 85% or less, 85% or more, 90% or less, 90% or more, 95% or more, 95% or more, or 100%, and "or less" can be interpreted as "less than," and "or greater than or equal to," can be interpreted as "greater than." These upper and lower limits can be combined as desired. In the present invention, the "ratio of malt used" refers to the ratio of the mass of malt to the mass of all raw materials excluding hops and water.

[0014] In the beer-taste non-alcoholic beverage of the present invention, in addition to malt, ungerminated wheat species such as ungerminated barley (including extracts) and ungerminated wheat (including extracts) can also be used as raw materials.

[0015] The beer-taste non-alcoholic beverage of the present invention is characterized in that the concentration of peptides with molecular weights of 800 to 4600 Da (gel filtration for HPLC analysis) in the beverage fractionated by gel filtration for HPLC analysis is within a predetermined range. In the present invention, the peptides with molecular weights of 800 to 4600 Da are derived from a beer-taste beverage made at least in part from malt, and are preferably derived from a fermented malt beverage (or fermented wort broth).

[0016] In the non-alcoholic beer-flavored beverage of the present invention, the lower limit (or higher) of the concentration of peptides with a molecular weight of 800 to 4,600 Da (gel filtration method for HPLC analysis) in the beverage is 0.25 mg / mL, but the lower limit (or higher) of the concentration of peptides with a molecular weight of 800 to 4,600 Da (gel filtration method for HPLC analysis) in the beverage is 0.26 mg / mL, 0.27 mg / mL, 0.28 mg / mL, 0.29 mg / mL, 0.30 mg / mL, 0.31 mg / mL, 0.32 mg / mL, 0.33 mg / mL, 0.34 mg / mL, 0.35 mg / mL, 0.36 mg / mL, 0.37 mg / mL, 0.38 mg / mL, 0.39 mg / mL, 0.40 mg / mL, 0.41 mg / mL, g / mL, 0.42 mg / mL, 0.43 mg / mL, 0.44 mg / mL, 0.45 mg / mL, 0.46 mg / mL, 0.47 mg / mL, 0.48 mg / mL, 0.49 mg / mL, 0.50 mg / mL, 0.51 mg / mL, 0.52 mg / mL, 0.53 mg / mL , 0.54 mg / mL, 0.55 mg / mL, 0.56 mg / mL, 0.57 mg / mL, 0.58 mg / mL, 0.59 mg / mL, 0 .60mg / mL, 0.61mg / mL, 0.62mg / mL, 0.63mg / mL, 0.64mg / mL, 0.65mg / mL, 0.6 6mg / mL, 0.67mg / mL, 0.68mg / mL, 0.69mg / mL, 0.70mg / mL, 0.71mg / mL, 0.72m g / mL, 0.73 mg / mL, 0.74 mg / mL, 0.75 mg / mL, 0.76 mg / mL, 0.77 mg / mL, 0.78 mg / mL, 0.79 mg / mL, 0.80 mg / mL, 0.81 mg / mL, 0.82 mg / mL, 0.83 mg / mL, 0.84 mg / mL , 0.85mg / mL, 0.86mg / mL, 0.87mg / mL, 0.88mg / mL, 0.89mg / mL, 0.90mg / mL, 0 .. 91mg / mL, 0.92mg / mL, 0.93mg / mL, 0.94mg / mL, 0.95mg / mL, 0.96mg / mL, 0.97 mg / mL, 0.98 mg / mL, 0.99 mg / mL, 1.00 mg / mL, 1.01 mg / mL, 1.02 mg / mL, 1.03 mg / mL, 1.04mg / mL, 1.05mg / mL, 1.06mg / mL, 1.07mg / mL, 1.08mg / mL, 1.09mg / m L, 1.10 mg / mL, 1.11 mg / mL, 1.12 mg / mL, 1.13 mg / mL, 1.14 mg / mL, 1.15 mg / mL,It can also be set to 1.16 mg / mL, 1.17 mg / mL, 1.18 mg / mL, 1.19 mg / mL, 1.20 mg / mL, 1.21 mg / mL, 1.22 mg / mL, 1.23 mg / mL, 1.24 mg / mL, 1.25 mg / mL, 1.26 mg / mL, 1.27 mg / mL, 1.28 mg / mL, 1.29 mg / mL, 1.30 mg / mL, 1.31 mg / mL, 1.32 mg / mL, 1.33 mg / mL, 1.34 mg / mL, 1.35 mg / mL, 1.36 mg / mL, 1.37 mg / mL, 1.38 mg / mL, 1.39 mg / mL or 1.40 mg / mL. The upper limit value (hereinafter or less) is 3.00 mg / mL, but it can also be 2.99 mg / mL, 2.98 mg / mL, 2.97 mg / mL, 2.96 mg / mL, 2.95 mg / mL, 2.94 mg / mL, 2.93 mg / mL, 2.92 mg / mL, 2.91 mg / mL, 2.90 mg / mL, 2.89 mg / mL, 2.88 mg / mL, 2.87 mg / mL, 2.86 mg / mL, 2.85 mg / mL, 2.84 mg / mL, 2.83 mg / mL, 2.82 mg / mL, 2.81 mg / mL, 2.80 mg / mL, 2.79 mg / mL, 2.78 mg / mL, 2.77 mg / mL, 2.76 mg / mL, 2.75 mg / mL, 2.74 mg / mL, 2.73 mg / mL, 2.72 mg / mL, 2.71 mg / mL, 2.70 mg / mL, 2.69 mg / mL, 2.68 mg / mL, 2.67 mg / mL, 2.66 mg / mL, 2.65 mg / mL, 2.64 mg / mL, 2.63 mg / mL, 2.62 mg / mL, 2.61 mg / mL, 2.60 mg / mL, 2.59 mg / mL, 2.58 mg / mL, 2.57 mg / mL, 2.56 mg / mL, 2.55 mg / mL, 2.54 mg / mL, 2.53 mg / mL, 2.52 mg / mL, 2.51 mg / mL, 2.50 mg / mL, 2.49 mg / mL, 2.48 mg / mL, 2.47 mg / mL, 2.46 mg / mL, 2.45 mg / mL, 2.44 mg / mL, 2.43 mg / mL, 2.42 mg / mL, 2.41 mg / mL, 2.40 mg / mL, 2.39 mg / mL, 2.38 mg / mL, 2.37 mg / mL, 2.36 mg / mL, 2.35 mg / mL, 2.34 mg / mL, 2.33 mg / mL, 2.32 mg / mL, 2.31 mg / mL, 2.30 mg / mL, 2.29 mg / mL,It can also be 2.28 mg / mL, 2.27 mg / mL, 2.26 mg / mL, 2.25 mg / mL, 2.24 mg / mL, 2.23 mg / mL, 2.22 mg / mL, 2.21 mg / mL, 2.20 mg / mL, 2.19 mg / mL, 2.18 mg / mL, 2.17 mg / mL, 2.16 mg / mL, 2.15 mg / mL, 2.14 mg / mL, 2.13 mg / mL, 2.12 mg / mL, 2.11 mg / mL, 2.10 mg / mL, 2.09 mg / mL, 2.08 mg / mL, 2.07 mg / mL, 2.06 mg / mL, 2.05 mg / mL, 2.04 mg / mL, 2.03 mg / mL, 2.02 mg / mL, 2.01 mg / mL or 2.00 mg / mL. These lower and upper limits can be combined in any desired manner, and the concentration of peptides having a molecular weight of 800 to 4,600 Da (as determined by gel filtration for HPLC analysis) in the beer-flavored non-alcoholic beverage of the present invention may be, for example, 0.25 mg / mL to 3.00 mg / mL, 0.30 mg / mL to 3.00 mg / mL, 0.35 mg / mL to 3.00 mg / mL, 0.40 mg / mL to 3.00 mg / mL, 0.45 mg / mL to 3.00 mg / mL, 0.50 mg / mL to 3.00 mg / mL, 0.60 mg / mL to 3.00 mg / mL, 0.70 mg / mL to 3.00 mg / mL, 0.80 mg / mL to 3.00 mg / mL, 0.90 mg / mL to 3.00 mg / mL, or 1.00 mg / mL to 3.00 mg / mL. Bottom, 1.10 mg / mL or more and 3.00 mg / mL or less, 1.20 mg / mL or more and 3.00 mg / mL or less, 1.30 mg / mL or more and 3.00 mg / mL or less, 1.40 mg / mL or more and 3.00 mg / mL or less, 0.25 mg / mL or more and 2.70 mg / mL or less, 0.30 mg / mL or more and 2.70 mg / mL or less, 0.35 mg / mL or more and 2.70 mg / mL or less, and 0.40 mg / mL or less Upper 2.70 mg / mL or less, 0.45 mg / mL or more and 2.70 mg / mL or less, 0.50 mg / mL or more and 2.70 mg / mL or less, 0.60 mg / mL or more and 2.70 mg / mL or less, 0.70 m g / mL or more and 2.70 mg / mL or less, 0.80 mg / mL or more and 2.70 mg / mL or less, 0.90 mg / mL or more and 2.70 mg / mL or less, 1.00 mg / mL or more and 2.70 mg / mL or less,1.10 mg / mL to 2.70 mg / mL, 1.20 mg / mL to 2.70 mg / mL, 1.30 mg / mL to 2.70 mg / mL, 1.40 mg / mL to 2.70 mg / mL, 0.25 mg / mL to 2.40 mg / mL Below, 0.30mg / mL to 2.40mg / mL, 0.35mg / mL to 2.40mg / mL, 0.40mg / mL to 2.40mg / mL, 0.45mg / mL to 2.40mg / mL, 0.50mg / mL to 2.40mg / mL or less, 0.60 mg / mL or more and 2.40 mg / mL or less, 0.70 mg / mL or more and 2.40 mg / mL or less, 0.80 mg / mL or more and 2.40 mg / mL or less, 0.90 mg / mL or more and 2.40 mg / mL or less, 1.00 mg / mL or more and 2.40 mg / mL or less, 1.10 mg / mL or more and 2.40 mg / mL or less, 1.20 mg / mL or more and 2.40 mg / mL or less, 1.30 mg / mL or more and 2.40 mg / mL or less, or 1.40 mg / mL or more and 2.40 mg / mL or less.

[0017] The beer-taste non-alcoholic beverage of the present invention is characterized in that the ratio (percentage) of the mass of peptides in the beverage having a molecular weight of 800 to 4,600 Da (gel filtration for HPLC analysis) to the mass of all peptides derived from the beverage fractionated by gel filtration for HPLC analysis is within a predetermined range. This ratio can be calculated by dividing the concentration (mg / mL) of peptides in the beverage having a molecular weight of 800 to 4,600 Da (gel filtration for HPLC analysis) by the concentration (mg / mL) of all peptides derived from the beverage fractionated by gel filtration for HPLC analysis.In the present invention, the lower limit of the peptide ratio (greater than or equal to) is 25.0%, but also includes 25.1%, 25.2%, 25.3%, 25.4%, 25.5%, 25.6%, 25.7%, 25.8%, 25.9%, 26.0%, 26.1%, 26.2%, 26.3%, 26.4%, 26.5%, 26.6%, 26.7%, 26.8%, 26.9%, 27.0, 27.1%, 27.2%, 27.3%, 27.4%, 27.5%, 27.6%, 27.7%, and 27.8%. ,27.9%,28.0%,28.1%,28.2%,28.3%,28.4%,28.5%,28.6%,28.7%,28.8%,28.9%,29.0,29.1%,29.2%,29.3%,29.4%,29.5%,29.6%,29.7%,29.8%,29.9%,30.0%,30.1%,30.2%,30.3%,30.4%,30.5%,30.6%,30.7%,30.8%,30.9%,31.0%,31.1%,31.2%,31.3 %, 31.4%, 31.5%, 31.6%, 31.7%, 31.8%, 31.9%, 32.0%, 32.1%, 32.2%, 32.3%, 32.4%, 32.5%, 32.6%, 32.7%, 32.8%, 32.9%, 33.0%, 33.1%, 33.2%, 33.3%, 33.4%, 33.5%, 33.6%, 33.7%, 33.8%, 33.9%, 34.0%, 34.1%, 34.2%, 34.3%, 34.4%, 34.5%, 34.6%, 34.7%, 3 It can also be 4.8%, 34.9%, 35.0%, 35.1%, 35.2%, 35.3%, 35.4%, 35.5%, 35.6%, 35.7%, 35.8%, 35.9%, 36.0%, 36.1%, 36.2%, 36.3%, 36.4%, 36.5%, 36.6%, 36.7%, 36.8%, 36.9%, 37.0%, 37.1%, 37.2%, 37.3%, 37.4%, 37.5%, 37.6%, 37.7%, 37.8%, 37.9% or 38.0%.The upper limit (less than or equal to) is 50.0%, but the following values ​​are also available: 49.9%, 49.8%, 49.7%, 49.6%, 49.5%, 49.4%, 49.3%, 49.2%, 49.1%, 49.0%, 48.9%, 48.8%, 48.7%, 48.6%, 48.5%, 48.4%, 48.3%, 49.4%. 8.2%, 48.1%, 48.0%, 47.9%, 47.8%, 47.7%, 47.6%, 47.5%, 47.4%, 47.3%, 47.2%, 47.1%, 47.0%, 46.9%, 46.8%, 46.7%, 46.6%, 46.5%, 46.4%, 46.3%, 46.2%, 46.1 %, 46.0%, 45.9%, 45.8%, 45.7%, 45.6%, 45.5%, 45.4%, 45.3%, 45.2%, 45.1%, 45.0%, 44.9%, 44.8%, 44.7%, 44.6%, 44.5%, 44.4%, 44.3%, 44.2%, 44.1%, 44.0%, 4 It can also be 3.9%, 43.8%, 43.7%, 43.6%, 43.5%, 43.4%, 43.3%, 43.2%, 43.1%, 43.0%, 42.9%, 42.8%, 42.7%, 42.6%, 42.5%, 42.4%, 42.3%, 42.2%, 42.1% or 42.0%.These lower and upper limits can be combined in any desired manner, and the peptide ratio in the beer-flavored non-alcoholic beverage of the present invention can range, for example, from 25.0% to 50.0%; from 30.0% to 50.0%, from 31.0% to 50.0%, from 32.0% to 50.0%, from 33.0% to 50.0%, from 34.0% to 50.0%, from 35.0% to 50.0%, from 36.0% to 50.0%, from 37.0% to 50.0%, or from 38.0% to 50.0%. 50.0% or less, 30.0% to 45.0%, 31.0% to 45.0%, 32.0% to 45.0%, 33.0% to 45.0%, 34.0% to 45.0%, 35.0% to 45.0%, 36.0% to 45.0% 37.0% to 45.0%, 38.0% to 45.0%, 30.0% to 44.0%, 31.0% to 44.0%, 32.0% to 44.0%, 33.0% to 44.0%, 34.0% to 44.0%, 35. 0% to 44.0%, 36.0% to 44.0%, 37.0% to 44.0%, 30.0% to 43.0%, 31.0% to 43.0%, 32.0% to 43.0%, 33.0% to 43.0%, 34.0% to 4 3.0% or less, 35.0% or more and 43.0% or less, 36.0% or more and 43.0% or more, 37.0% or more and 43.0% or less, 30.0% or more and 42.5% or less, 31.0% or more and 42.5% or less, 32.0% or more and 42.5% or less, 33.0% or more and 42.5% or less or lower, 34.0% or more and 42.5% or less, 35.0% or more and 42.5% or less, 36.0% or more and 42.5% or less, 37.0% or more and 42.5% or less, 38.0% or more and 42.5% or less, 30.0% or more and 42.0% or less, 31.0% or more and 42.0% or less, 32.0% or more and 42.0% or less, 33.0% or more and 42.0% or less, 34.0% or more and 42.0% or less, 35.0% or more and 42.0% or less, 36.0% or more and 42.0% or less, 37.0% or more and 42.0% or less, or 38.0% or more and 42.0% or less.

[0018] The molecular weight of peptides in beverages is measured by gel filtration using high-performance liquid chromatography (sometimes referred to herein as "gel filtration for HPLC analysis"). Specifically, molecular weights are calculated from retention times using a calibration curve for gel filtration for HPLC analysis (see, for example, Example 1(1)A(vi) and Figure 1 in the Examples below), and samples with different molecular weights can be separated based on their retention times. For specific examples of molecular weight measurement using gel filtration for HPLC analysis, see, for example, Example 1(1)A(iii) to (vi) in the Examples below. Furthermore, peptide quantification can be performed by the Lowry method.

[0019] The beer-flavored non-alcoholic beverage of the present invention is characterized by a suppressed sweet taste and aroma and a beer-like balance, despite being a non-alcoholic beverage. Problems with beer-flavored non-alcoholic beverages include a strong sweet taste and aroma and a poor beer-like balance due to the non-alcoholic nature of the beverage and the unique method used to prepare it. The beer-flavored non-alcoholic beverage of the present invention suppresses the sweet taste and aroma and achieves a beer-like balance, thereby solving the above-mentioned problems of beer-flavored non-alcoholic beverages. Here, "sweet taste and aroma" refers to a flavor in which the sweetness of the taste and aroma is strong, and the sweetness lingers after swallowing, resulting in a lack of crispness. Furthermore, "beer-like balance" refers to a flavor in which the taste elements (sweetness, bitterness, sourness, umami, etc.) that make up the natural taste of beer are not prominent, but are instead well-balanced and beer-like.

[0020] The beer-flavored non-alcoholic beverage of the present invention can be produced according to a typical production procedure for a beer-flavored non-alcoholic beverage, so long as the peptides are added so that the concentration of the 800 to 4600 Da peptides in the beverage falls within a predetermined range. The beer-flavored non-alcoholic beverage of the present invention can be produced, for example, by preparing a brewing liquid from malt, hops, water, and other ingredients, allowing the brewing liquid to stand to remove solids, and then filtering the brewing liquid. The other ingredients may be used as ingredients for the brewing liquid, or may be blended with the prepared brewing liquid. This method allows the content of the 800 to 4600 Da peptides to be adjusted at any stage.

[0021] The preparation of the brewing liquid can be carried out according to a conventional method, for example, by sequentially carrying out the following steps: (a) saccharifying a mixture of water and raw materials containing malt, followed by filtering to obtain wort; (b) adding hops to the obtained wort and then boiling the wort; and (c) cooling the boiled wort.

[0022] In the present invention, wort can also be prepared by adding commercially available enzyme preparations during the saccharification step. For example, a protease preparation can be used for protein degradation, an α-amylase preparation, a glucoamylase preparation, a pullulanase preparation, or the like for carbohydrate degradation, and a β-glucanase preparation, a cellulose-degrading enzyme preparation, or the like for cellulose degradation, or a mixture of these preparations can also be used.

[0023] In the present invention, malt with high endoprotease activity and / or an enzyme preparation possessing endoprotease activity can be used in the saccharification step to promote proteolysis, and malt with low endoprotease activity can be used to inhibit proteolysis. By carrying out such steps, the concentration of peptides with a molecular weight of 800 to 4,600 Da (as determined by gel filtration for HPLC analysis) contained in a beer-flavored non-alcoholic beverage can be adjusted to fall within a predetermined range, thereby producing a beer-flavored non-alcoholic beverage that has a reduced sweet taste and aroma and a well-balanced flavor that is characteristic of beer, despite being a non-alcoholic beverage.

[0024] In the present invention, the saccharification temperature can be, for example, 35°C to 100°C. From the viewpoint of adjusting the content of 800 to 4600 Da peptides to a predetermined numerical range, it can be preferably 35°C to 100°C, more preferably 40°C to 100°C, and even more preferably 45°C to 100°C. The saccharification time can be appropriately set taking into account the saccharification temperature, and can be, for example, 5 minutes to 120 minutes. When the saccharification temperature is 45°C to 100°C, it can be preferably 10 minutes to 120 minutes (more preferably 30 minutes to 120 minutes). In the present invention, the boiling conditions can be set according to conventional methods.

[0025] The beer-taste non-alcoholic beverage of the present invention can also be produced, for example, by producing a fermented malt beverage such as beer and then removing alcohol from the resulting fermented malt beverage, or by terminating fermentation in the fermentation step (corresponding to step (III) described below) of the production of a fermented malt beverage at a stage where the ethanol concentration is less than 1 v / v %. This method allows the content of 800 to 4600 Da peptides to be adjusted at any stage of the production process.

[0026] The fermented malt beverage used as the raw material for the beer-flavored non-alcoholic beverage of the present invention can be produced according to a conventional production procedure. For example, a fermented malt beverage can be produced by sequentially performing the following steps: (I) saccharifying a mixture of malt-containing raw materials and warm water, followed by filtering to obtain wort; (II) adding hops to the resulting wort and then boiling it; (III) cooling the boiled wort; (IV) adding yeast to the cooled wort and fermenting it; and (V) storing the resulting wort fermentation liquid at low temperature and then removing the yeast by filtration. As described above, wort can also be produced by adding a commercially available enzyme preparation during the saccharification step. The saccharification conditions can be set as described above, and the boiling, fermentation, and storage conditions can be set according to conventional methods.

[0027] Methods for removing alcohol from the above-mentioned fermented malt beverages include, for example, (i) a method for removing alcohol and low-boiling point components by distillation under reduced pressure or normal pressure, (ii) a method for removing alcohol and low-molecular-weight components using a reverse osmosis (RO) membrane, and (iii) a method for removing volatile components by adsorbing them into steam using centrifugal force.

[0028] In the production of the beer-flavored non-alcoholic beverage of the present invention, in addition to malt and ungerminated wheat and barley, rice, corn, soybeans, koryang (Japanese radish), potatoes, starch, sugars (e.g., liquid sugar), fruits (e.g., fruit juice, concentrated fruit juice), coriander or its seeds, spices or raw materials thereof (e.g., pepper, cinnamon, cloves, Japanese pepper), herbs (e.g., chamomile, sage, basil, lemongrass), vegetables (e.g., sweet potato, pumpkin), buckwheat or sesame, carbohydrate-containing substances (e.g., honey, brown sugar), salt, miso, flowers, tea, coffee, cocoa (tea, coffee, and cocoa include preparations thereof), seafood (e.g., Other additives that can be used as raw materials include: oysters, kelp, wakame seaweed, and dried bonito flakes), nitrogen sources such as protein hydrolysates and yeast extract, hops or hop processed products (e.g., hop extract), flavorings (e.g., commercially available beer flavors containing ethyl acetate, isoamyl acetate, and isoamyl alcohol, which are typical aroma components of beer), coloring components (e.g., colorings such as caramel color), foaming and foam retention improvers, sweeteners (e.g., high-intensity sweeteners), seasoning components (e.g., amino acids), acidulants (e.g., gluconic acid), bittering components (e.g., naringin), dietary fiber, herbs, antioxidants, and water quality conditioners. The content of dietary fiber (e.g., indigestible dextrin) in the beer-flavored non-alcoholic beverage of the present invention can be less than 1% (preferably less than 0.9%).

[0029] When producing the fermented beer-taste non-alcoholic beverage of the present invention, for example, one or more of the above-mentioned ingredients can be used as raw materials for producing the fermented malt beverage in addition to malt and hops. In producing the fermented beer-taste non-alcoholic beverage of the present invention, one or more of the above-mentioned ingredients can be blended during the production process of the fermented malt beverage, or one or more of the above-mentioned ingredients can be blended before or after removal of alcohol after production of the fermented malt beverage.

[0030] When producing the non-fermented, beer-taste, non-alcoholic beverage of the present invention, for example, one or more of the above-mentioned ingredients can be used in addition to malt and hops as raw materials for producing wort. In producing the non-fermented, beer-taste, non-alcoholic beverage of the present invention, one or more of the above-mentioned ingredients can be blended during the wort production process, or one or more of the above-mentioned ingredients can be blended after wort production.

[0031] In the present invention, a fraction containing peptides with a molecular weight of 800 to 4600 Da is prepared from malt and added to a beer-taste non-alcoholic beverage. This allows the concentration of peptides with a molecular weight of 800 to 4600 Da in the beverage to be adjusted to a predetermined range, thereby producing a beer-taste non-alcoholic beverage with a suppressed sweet taste and aroma and a beer-like balance. Specifically, the present invention provides a method for producing a beer-taste non-alcoholic beverage, which comprises preparing a fraction containing peptides with a molecular weight of 800 to 4600 Da (HPLC analytical gel filtration method) from malt and adding the fraction to a beer-taste non-alcoholic beverage. The peptide fraction can be added to the beer-taste non-alcoholic beverage during the beverage's production process (e.g., the blending process).

[0032] The present invention also provides a method for producing a beer-taste non-alcoholic beverage, comprising producing a fermented malt beverage using malt as at least a part of the raw material, preparing a fraction from the beverage containing peptides with a molecular weight of 800 to 4600 Da, and adding the fraction to a beer-taste non-alcoholic beverage, thereby adjusting the concentration of peptides with a molecular weight of 800 to 4600 Da in the beverage to fall within a predetermined range, thereby producing a beer-taste non-alcoholic beverage with a suppressed sweet taste and aroma and a beer-like balance. Specifically, the present invention provides a method for producing a beer-taste non-alcoholic beverage, comprising producing a fermented malt beverage using malt as at least a part of the raw material, preparing a fraction from the beverage containing peptides with a molecular weight of 800 to 4600 Da (as determined by gel filtration for HPLC analysis), and adding the fraction to a beer-taste non-alcoholic beverage. The present invention also provides a method for producing a non-alcoholic beer-taste beverage, which comprises preparing a fraction containing peptides with a molecular weight of 800 to 4,600 Da (as determined by gel filtration for HPLC analysis) from a fermented malt beverage made at least in part from malt, and adding the fraction to a non-alcoholic beer-taste beverage. The peptide fraction can be added to the non-alcoholic beer-taste beverage during the production process (e.g., the blending process) of the beverage.

[0033] The present invention also provides a beer-flavored non-alcoholic beverage containing one or more peptides with a molecular weight of 800 to 4,600 Da derived from a fermented malt beverage containing malt as at least a portion of its raw material, which beverage is a part of the beer-flavored non-alcoholic beverage of the present invention. The beverage containing the peptide has an improved or enhanced flavor as a beer-flavored non-alcoholic beverage, specifically, a beverage with a reduced sweet taste and aroma and a well-balanced flavor that is characteristic of beer.

[0034] The carbohydrate concentration of the beer-flavored non-alcoholic beverage of the present invention is not particularly limited, but the lower limit (not less than or exceeding) can be 0.5 g / 100 mL or 1.0 g / 100 mL, and the upper limit (not more than or less than) can be 7.0 g / 100 mL, 6.95 g / 100 mL, 6.9 g / 100 mL, 6.85 g / 100 mL, 6.8 g / 100 mL, 6.75 g / 100 mL, The upper limit and lower limit may be less than 6.7 g / 100 mL, 6.65 g / 100 mL, 6.6 g / 100 mL, 6.55 g / 100 mL, 6.5 g / 100 mL, 6.4 g / 100 mL, 6.3 g / 100 mL, 6.2 g / 100 mL, 6.1 g / 100 mL, 6.0 g / 100 mL, 5.5 g / 100 mL, or 5.0 g / 100 mL, and these upper and lower limits may be combined arbitrarily.

[0035] The carbohydrate concentration can be measured by known methods, and can be calculated by subtracting the amounts of water, protein, fat, ash, and dietary fiber from the mass of the sample to be measured (see the Nutrition Labeling Standards (Consumer Affairs Agency Notification No. 9, December 16, 2009, partial revision)).

[0036] The concentration of the true (genuine) extract in the non-alcoholic beer-flavored beverage of the present invention is not limited, but the lower limit (or higher) thereof is 0.3 w / w%, 0.4 w / w%, 0.5 w / w%, 0.6 w / w%, 0.7 w / w%, 0.8 w / w%, 0.9 w / w%, 1.0 w / w%, 1.1 w / w%, 1.2 w / w%, 1.3 w / w%, 1.4 w / w%, 1.5 w / w%, 1.6 w / w%, 1.7 w / w%, 1.8 w / w%, 1.9 w / w%, 2.0 w / w%, 2.1 w / w%, 2.2 w / w%, 2.3 w / w%, 2.4 w / w%, 2.5 w / w%, 2.6 w / w%, 2.7 w / w%, 2.8 w / w%, 2 ... . 1 w / w%, 2.2 w / w%, 2.3 w / w%, 2.4 w / w%, 2.5 w / w%, 2.6 w / w%, 2.7 w / w%, 2.8 w / w%, 2.9 w / w%, 3.0 w / w%, 3.1 w / w%, 3.2 w / w%, 3.3 w / w%, 3.4 w / w%, 3.5 w / w%, 3.6 w / w%, 3.7 w / w%, 3.8 w / w%, 3.9 w / w% or 4.0 w / w%, and the upper limit (not more than or less than) is 10 w / w%, 9.9 w / w%, 9.8 w / w%, 9. 7w / w%, 9.6w / w%, 9.5w / w%, 9.4w / w%, 9.3w / w%, 9.2w / w%, 9.1w / w%, 9.0w / w%, 8.9w / w%, 8.8w / w%, 8.7w / w%, 8.6w / w%, 8.5w / w%, 8.4 w / w%, 8.3w / w%, 8.2w / w%, 8.1w / w%, 8.0w / w%, 7.9w / w%, 7.8w / w%, 7.7w / w%, 7.6w / w%, 7.5w / w%, 7.4w / w%, 7.3w / w%, 7.2w / w%, 7.1w %, 7.0 w / w%, 6.9 w / w%, 6.8 w / w%, 6.7 w / w%, 6.6 w / w%, 6.5 w / w%, 6.4 w / w%, 6.3 w / w%, 6.2 w / w%, 6.1 w / w%, 6.0 w / w%, 5.9 w / w%, 5.8 w / w%, 5.7 w / w%, 5.6 w / w%, 5.5 w / w%, 5.4 w / w%, 5.3 w / w%, 5.2 w / w%, 5.1 w / w%, and 5.0 w / w%, and these upper and lower limits can be combined arbitrarily. The true extract concentration can be measured at 20°C using a commercially available device (e.g., Alcolyzer (Anton Paar)).

[0037] The pH (measured at 25°C) of the beer-taste non-alcoholic beverage of the present invention is not limited, but the lower limit (greater than or equal to) can be 3.5, 3.6, 3.7, or 3.8, and the upper limit (less than or equal to) can be 4.6, 4.5, 4.4, 4.35, or 4.3. These lower and upper limits can be combined arbitrarily, for example, 3.5 to 4.6, 3.5 to 4.5, 3.5 to 4.4, or 3.5 to less than 4.4. The pH of the beer-taste non-alcoholic beverage of the present invention can be adjusted using a pH adjuster. The pH of the beer-taste non-alcoholic beverage can be measured using a commercially available pH meter (e.g., a benchtop pH meter, manufactured by Horiba, Ltd.).

[0038] The EBC color score of the beer-taste non-alcoholic beverage of the present invention is not limited, but the lower limit (not less than or exceeding) can be 4.0 EBC, 4.5 EBC, 5.0 EBC, 5.5 EBC, or 6.0 EBC, and the upper limit (not more than or less than) can be 10.0 EBC, 9.9 EBC, 9.8 EBC, 9.7 EBC, 9.6 EBC, 9.5 EBC, 9.4 EBC, 9.3 EBC, or 10.0 EBC. The EBC color index can be 8.9EBC, 8.8EBC, 8.7EBC, 8.6EBC, 8.5EBC, 8.4EBC, 8.3EBC, 8.2EBC, 8.1EBC, 8.0EBC, 7.9EBC, 7.8EBC, 7.7EBC, 7.6EBC, 7.5EBC, 7.4EBC, 7.3EBC, 7.2EBC, 7.1EBC, or 7.0EBC. These lower and upper limits can be combined arbitrarily. The EBC color index of the beer-flavored non-alcoholic beverage of the present invention can be measured using "Revised BCOJ Beer Analysis Method 4.3.8, compiled by the International Technical Committee (Analysis Committee) of the Brewers Association of Japan, Brewery Society of Japan."

[0039] The bitterness value (BU) of the beer-taste non-alcoholic beverage of the present invention is not limited, but the lower limit (not less than or exceeding) can be 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, or 6.0, and the upper limit (not more than or less than) can be 23.0, 22.9, 22.8, 22.7, 22.6, 22.5, 22.4, 22.3, 22.2, 22.1, 22.0, 21.9, 21.8, 21.7, 21.8, 21.9 ... .6, 21.5, 21.4, 21.3, 21.2, 21.1, 21.0, 20.9, 20.8, 20.7, 20.6, 20.5, 20.4, 20.3, 20.2, 20.1, 20.0, 19.9, 19.8, 19.7, 19.6, 19.5, 19.4, 19.3, 19.2, 19.1, 19.0, 18.9, 18.8, 18.7, 18.6, 18.5, 18.4, 18.3, 18.2, 18.1 or 18.0. These lower and upper limits can be arbitrarily combined, for example, 2.0 to 23.0, 3.0 to 23.0, 3.5 to 22.0, 4.0 to 22.0, or 5.0 to 21.0. In the present invention, "bitterness value (BU)" refers to a value measured according to the bitterness value method described in "8.15" of the BCOJ Beer Analysis Method (revised and expanded in 2013). Specifically, the bitterness value can be determined by adding an acid to a sample beverage, extracting it with isooctane, measuring the absorbance (275 nm) of the isooctane layer, and multiplying this measured value by 50.

[0040] The beverage provided by the present invention can be provided as a bottled beverage after optionally being subjected to a step of adding carbon dioxide gas, and further undergoing steps such as a filling step and a sterilization step. Sterilization may be carried out either before or after filling into a container.

[0041] The beer-taste non-alcoholic beverage of the present invention can be provided as a carbonated beverage. The carbon dioxide pressure (gas pressure at 20°C) of the beer-taste non-alcoholic beverage of the present invention is not limited, but the lower limit (greater than or equal to) can be 0.05 MPa, 0.06 MPa, 0.07 MPa, 0.08 MPa, 0.09 MPa, or 0.1 MPa, and the upper limit (less than or equal to) can be 0.4 MPa, 0.39 MPa, 0.38 MPa, 0.37 MPa, 0.36 MPa, or 0.35 MPa. These upper and lower limits can be combined in any manner, and can be, for example, 0.05 MPa to 0.4 MPa, 0.07 MPa to 0.38 MPa, or 0.1 MPa to 0.35 MPa.

[0042] The container used for the beverage of the present invention may be any container that is normally used for filling beverages, such as a metal can, a barrel, a plastic bottle (e.g., a PET bottle or cup), a paper container, a bottle, or a pouch container, but metal cans, barrels, plastic bottles (e.g., a PET bottle), or a bottle are preferred.

[0043] According to another aspect of the present invention, there is provided a method for producing a flavor-improved non-alcoholic beer-taste beverage made at least in part from malt, the method comprising the step of incorporating peptides having molecular weights of 800 to 4600 Da (as fractionated by HPLC gel filtration) into the beverage so that the concentration of said peptides in the beverage is 0.25 mg / mL or more and 3.00 mg / mL or less. In the present invention, "improved flavor" or "flavor improvement" of a non-alcoholic beer-taste beverage refers to achieving a beer-like balance while suppressing the sweet taste and aroma of the non-alcoholic beer-taste beverage. The above-described production method of the present invention is preferably a method for producing a beer-taste non-alcoholic beverage that suppresses the sweet taste and aroma and achieves a beer-like balance. The above-described production method of the present invention can be carried out according to the description of the non-alcoholic beer-taste beverage of the present invention.

[0044] According to another aspect of the present invention, there is provided a flavor improver for a beer-taste non-alcoholic beverage, comprising as an active ingredient a peptide having a molecular weight of 800 to 4,600 Da (as determined by gel filtration for HPLC analysis) contained in a beer-taste beverage (preferably a fermented malt beverage) made at least in part from malt. The flavor improver of the present invention can be produced according to the description of the beer-taste non-alcoholic beverage and the method for producing the same of the present invention.

[0045] According to yet another aspect of the present invention, there is provided a method for improving the flavor of a beer-taste non-alcoholic beverage that uses malt as at least one raw material. The flavor improvement method of the present invention can be carried out by a step of incorporating peptides having a molecular weight of 800 to 4,600 Da (HPLC analytical gel filtration method) into the beverage fractionated by HPLC analytical gel filtration so that the concentration of the peptides is 0.25 mg / mL or more and 3.00 mg / mL or less. The flavor improvement method of the present invention can be carried out according to the description of the beer-taste non-alcoholic beverage of the present invention and the method for producing the same.

[0046] The present invention will be described in more detail based on the following examples, but the present invention is not limited to these examples.

[0047] Measurement of alcohol concentration The alcohol concentration (ethanol concentration) in the beverages was measured using gas chromatography (GC) with an FID detector (Agient). A calibration curve was prepared based on the concentrations of the target samples ranging from 0.000 to 0.05% v / v.

[0048] Measurement of True Extract Concentration The true extract concentration was measured at 20°C using an Alcolyzer (Anton Paar).

[0049] Example 1: Preparation and evaluation of beer-flavored non-alcoholic beverages In Example 1, test samples of beer-flavored non-alcoholic beverages were prepared and evaluation tests were carried out on each sample.

[0050] (1) Method A Preparation of Test Samples (i) Production of Dealcoholized Wort Fermentation Liquid Barley malt, enzymes, and warm water were placed in a mash tank and saccharified at a temperature of 60°C to 78°C. This saccharification liquid was filtered, transferred to a boiling kettle, and hops were added. The mixture was then boiled for 70 minutes. After boiling, the evaporated warm water was added to the resulting mixture, and the heat trub was removed in a whirlpool tank. The mixture was then cooled to 10°C to obtain cold wort. Brewer's yeast was added to the wort, and the mixture was fermented at around 10°C for 7 days, after which the brewer's yeast was removed. The resulting mixture was transferred to another tank and aged for 7 days, then cooled to around -1°C and stabilized for 14 days. The resulting mixture was then diluted with degassed water and filtered to obtain a wort fermentation liquid. The mixture was then sprayed into a degassing tank to remove carbon dioxide, and the mixture was then heated to around 50°C. The mixture was then brought into contact with steam heated to around 50°C in a reduced pressure column at around 60 mbar, causing the volatile components to be adsorbed by the steam, removing the alcohol and volatile components, and adding carbon dioxide gas to obtain a dealcoholized wort fermentation liquid (dealcoholized wort fermentation liquid) with an alcohol concentration of 0.001 v / v% (malt usage ratio 100%).

[0051] (ii) Production of wort Barley malt, enzymes, and warm water were added to a mash tank and saccharified. The saccharified liquid was filtered and transferred to a boiling kettle, where hops were added and boiled for 90 minutes. After boiling, warm water was added to the resulting mixture to compensate for the evaporation, and the mixture was placed in a whirlpool tank to remove the heat trough. The mixture was then cooled to obtain wort (100% malt content).

[0052] (iii) Fractionation by gel filtration for HPLC analysis: The dealcoholized wort fermentation broth obtained in (i) above was filtered through a 0.45 μm filter, and the filtered fermentation 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 fractionation under the following conditions.

[0053] <Gel filtration conditions for HPLC analysis> Column: Hiload Superdex 30pg26 / 600 (Cytiva) Sample injection volume: 5 mL Eluent composition: 100 mM NaCl Flow rate: 2.5 mL / min (constant flow rate) Detection wavelength: 215 nm Fractionation: 19.1 mL (fraction 0) from 0.29 cv (column volume), followed by 5 mL fractionation from 0.35 cv (fractions 1 to 51)

[0054] (iv) Purification of Peptide Fractions Fractions 8 to 28 obtained in (iii) above were subjected to adsorption treatment using a solid-phase extraction column (Bond Elute C18, Agilent Technologies), washed with demineralized water, eluted with 50% aqueous ethanol, concentrated to dryness, and reconstituted with demineralized water to obtain peptide fractions as a concentrate.

[0055] (v) Protein Quantification by Lowry Method: The peptide concentration of the peptide fraction obtained in (iii) above was quantified by the Lowry method using a commercially available kit (DC Protein Assay, Bio-Rad). First, the concentration of the fraction solution was adjusted to an appropriate range. 50 μL of solution A was added to 5 μL of the adjusted sample 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 and a previously prepared calibration curve. The calibration curve was prepared using BSA (bovine serum albumin). (vi) Calculation of Molecular Weight Range by Gel Filtration for HPLC Analysis: The molecular weight range corresponding to the peptide fractions (fractions 8-28) obtained in (iv) above was calculated by gel filtration for HPLC analysis. The specific procedure is as follows.

[0056] The peptides in the peptide fractions (fractions 8-28) were weighed and then lyophilized. This dried product was dissolved in 50 mM sodium phosphate buffer (pH 7.0, containing 150 mM NaCl) to prepare a 2.5-fold concentrated solution, which was used as the sample for fractionation. The sample was subjected to gel filtration fractionation under the following conditions. <Gel filtration conditions for HPLC analysis> Column: Superdex 75 increase (Cytiva) Sample injection volume: 100 μL Eluent composition: 50 mM sodium phosphate (pH 7.0), 20% (v / v) acetonitrile, 150 mM NaCl Flow rate: 0.5 mL / min (constant flow rate) Detection wavelength: 215 nm Fractionation program: shown in Table 1

[0057]

[0058] Using the column, eluent, flow rate, and detection wavelength described above for the gel filtration conditions for HPLC analysis, peptides with known molecular weights were dissolved in ultrapure water at concentrations of 0.1 to 5 mg / mL, and 50 μL of each solution was injected to perform gel filtration for HPLC analysis, and the retention times were confirmed (Table 2). A calibration curve was then created from the retention times and molecular weights, and the molecular weight range and fractionation range were determined (Figure 1).

[0059]

[0060] Each peptide fraction was analyzed using a Superdex 75 Increase (Cytiva) column to calculate the molecular weight. It was confirmed that the peptides in fractions 8 to 28 were distributed in the molecular weight range of 800 to 4600 Da, which corresponds to fractions #5 to #7.

[0061] (vii) Calculation of the ratio of peptides with a molecular weight of 800 to 4600 Da The ratio of peptides with a molecular weight of 800 to 4600 Da was calculated using the peptide concentration in each fraction equivalent to the product using the following calculation formula.

[0062] (viii) Preparation of Test Samples The test sample for Test Plot 1 was prepared by mixing the dealcoholized wort fermentation liquid obtained in (i) above with the wort obtained in (ii) above, diluting the mixture with water, and adding a liquid sugar mainly composed of non-assimilable sugars, so as to achieve the peptide concentrations shown in Table 3. On the other hand, the test samples for Test Plots 2 to 9 were prepared by adding the peptide fraction obtained in (iv) above (peptides of 800-4600 Da) to the test sample for Test Plot 1 so as to achieve the respective peptide concentrations shown in Table 3.

[0063] (i) Evaluation Test The test was conducted by five trained panelists. Specifically, the two evaluation items, "sweet taste and aroma" and "beer-like balance," were rated on a scale of 1 to 9 in increments of 0.5, and the average evaluation scores of the five panelists were calculated. Here, "sweet taste and aroma" refers to a flavor in which the taste and aroma are very sweet, and the sweetness lingers even after drinking, resulting in a lack of aftertaste. The higher the score, the more suppressed the sweet taste and aroma. "Beer-like balance" refers to a flavor in which the individual taste elements that make up the natural taste of beer (sweetness, bitterness, sourness, umami, etc.) are not prominent, and the higher the score, the more beer-like the balance. In addition, the score for each evaluation item in Test Plot 1 was set at 3.0, and Test Plots 2 to 9 were evaluated based on the sensory evaluation of Test Plot 1. In each evaluation item in Table 3, ◎ indicates a score of 5.5 or more and 9 or less, 〇 indicates a score of 3.5 or more and less than 5.5, and × indicates a score of 1 or more and less than 3.5. In the overall evaluation in Table 3, ◎ indicates that both evaluation items are ◎, 〇 indicates that the evaluation item is ◎ or 〇, and × indicates that one of the evaluation items is ×.

[0064] (2) Results The results are shown in Table 3. It was shown that when the concentration of 800 to 4600 Da peptides was at a predetermined value, the sweet taste and aroma were suppressed and a beer-like balance was achieved.

[0065]

[0066] Example 2: Production and evaluation of beer-flavored non-alcoholic beverages In Example 2, test samples of beer-flavored non-alcoholic beverages with various malt usage ratios were produced, and evaluation tests were carried out on each sample.

[0067] (1) Methods A. Preparation of test samples (i) Test area 1 Test samples for test area 1 were prepared in the same manner as in Example 1(1)A.

[0068] (ii) Test Plot 10 The test sample for Test Plot 10 was produced as a dealcoholized beverage with a 100% malt ratio. Specifically, barley malt, enzymes, and warm water were placed in a mash tank and saccharified at a temperature of 60°C to 78°C. The saccharified liquid was filtered, transferred to a boiling kettle, and hops were added. The mixture was then boiled for 70 minutes. After boiling, the evaporated warm water was added to the resulting mixture, trub was removed in a whirlpool tank, and the mixture was cooled to 10°C to obtain a cold wort. Brewer's yeast was added to the wort, and the mixture was fermented at around 10°C for 7 days, after which the brewer's yeast was removed. The resulting mixture was transferred to another tank and aged for 7 days, then cooled to around -1°C and stabilized for 14 days. The resulting mixture was then diluted with degassed water and filtered to obtain a fermented wort liquor. The mixture was then sprayed into a degassing tank to remove carbon dioxide, and then heated to around 50°C. The mixture was then brought into contact with steam heated to around 50°C in a reduced pressure column at around 60 mbar, the volatile components were adsorbed by the steam, the alcohol and volatile components were removed, and carbon dioxide gas was added to obtain a test sample of test area 10 with an alcohol concentration of 0.001 v / v%.

[0069] (ii) Test Area 11 The test sample for Test Area 11 was produced as a dealcoholic beverage with a 75% malt ratio. Specifically, saccharification was performed in a brewing tank using barley malt, rice, corn starch, and corn grits as raw materials. The saccharified liquid was filtered, transferred to a boiling kettle, and hops were added. The mixture was then boiled for 90 minutes. After boiling, hot water was added to the resulting mixture to remove the evaporated fluid. The mixture was then cooled to 9°C to obtain a cold wort. Brewer's yeast was added to the wort, and the mixture was fermented at around 10°C for 7 days, after which the brewer's yeast was removed. The resulting mixture was transferred to another tank and aged for 7 days, then cooled to around -1°C and stabilized for 14 days. The resulting mixture was then diluted with degassed water and filtered to obtain a fermented wort liquor. The mixture was then sprayed into a degassing tank to remove carbon dioxide, and then heated to around 50°C. The mixture was then brought into contact with steam heated to around 50°C in a reduced pressure column at around 60 mbar, the volatile components were adsorbed by the steam, the alcohol and volatile components were removed, and carbon dioxide gas was added to obtain a test sample of test area 11 with an alcohol concentration of 0.001 v / v%.

[0070] (i) Evaluation test An evaluation test was conducted in the same manner as in Example 1 (1) (i), and a comparison was made with Test Group 1 of Example 1. In addition, for each evaluation item in Table 4, ◎ indicates a score of 5.5 or more and 9 or less, 〇 indicates a score of 3.5 or more and less than 5.5, and × indicates a score of 1 or more and less than 3.5. In the overall evaluation in Table 4, ◎ indicates that both evaluation items are ◎, 〇 indicates that the evaluation item is ◎ or 〇, and × indicates that one of the evaluation items is ×.

[0071] (2) Results The results are shown in Table 4. It was shown that even in beer-flavored non-alcoholic beverages with 100% and 75% malt content, when the concentration of 800 to 4600 Da peptides was within a predetermined range, the sweet taste and aroma were suppressed and a beer-like balance was achieved.

[0072]

Claims

1. A non-alcoholic beer-taste beverage using malt as at least a part of the raw material, wherein the peptide concentration in the beverage fractionated by gel filtration for HPLC analysis and having a molecular weight of 800 to 4600 Da (gel filtration for HPLC analysis) is 0.25 mg / mL or more and 3.00 mg / mL or less.

2. The non-alcoholic beer-taste beverage according to claim 1, wherein the real extract concentration in the beverage is 0.3 w / w% or more and 10 w / w% or less.

3. The non-alcoholic beer-taste beverage according to claim 1 or 2, wherein the peptide concentration is 1.00 mg / mL or more and 3.00 mg / mL or less.

4. The non-alcoholic beer-taste beverage according to claim 1 or 2, wherein the malt usage ratio is 25% by mass or more.

5. The non-alcoholic beer-taste beverage according to claim 1 or 2, which is a de-alcoholized beverage.

6. A method for producing a non-alcoholic beer-taste beverage using malt as at least a part of the raw material, the method comprising a step of containing the peptide so that the peptide concentration in the beverage fractionated by gel filtration for HPLC analysis and having a molecular weight of 800 to 4600 Da (gel filtration for HPLC analysis) is 0.25 mg / mL or more and 3.00 mg / mL or less.

7. The production method according to claim 6, comprising a step of removing alcohol from the wort fermentation broth.

8. A method for improving the flavor of a non-alcoholic beer-taste beverage using malt as at least a part of the raw material, the method comprising, in the production method of the beverage, a step of containing the peptide so that the peptide concentration in the beverage fractionated by gel filtration for HPLC analysis and having a molecular weight of 800 to 4600 Da (gel filtration for HPLC analysis) is 0.25 mg / mL or more and 3.00 mg / mL or less.

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