JP2021106540A - Beer taste alcoholic beverage - Google Patents

Abstract

To provide a beer taste beverage that has excellent flavor stability while ensuring robust feel.SOLUTION: In a beer taste beverage, the content of a protein with a molecular weight of 35-50 kDa is 0.005 or more and 0.1 or less in a mass ratio to the total protein content.SELECTED DRAWING: None

Description

The present invention relates to a beer-taste beverage.

With the diversification of consumer tastes in recent years, the development of beer-taste beverages having various flavor characteristics is desired.

In beer-taste beverages, sugars and amino acids may be added to impart richness and the like. For example, Patent Document 1 discloses a non-fermented alcohol-taste beverage containing γ-aminobutyric acid (GABA).

Japanese Unexamined Patent Publication No. 2017-184697

However, since sugars and amino acids are highly reactive, the addition of these may cause a change in flavor during storage.

The present invention relates to providing a beer-taste beverage having excellent flavor stability while having a drinkable response.

The present invention relates to a beer-taste beverage in which the mass ratio of the protein content having a molecular weight of 35 to 50 kDa to the total protein content is 0.005 or more and 0.1 or less.

According to the present invention, it is possible to provide a beer-taste beverage having excellent flavor stability while having a drinkable response.

When the present inventors examined the above-mentioned problems, surprisingly, by setting the mass ratio of the protein content having a molecular weight of 35 to 50 kDa to the total protein content to a specific ratio, it was possible to have a drinkable response. It was newly found that a beer-taste beverage with excellent flavor stability can be obtained. Although the mechanism is not clear, it is presumed that the protein having a molecular weight of 35 to 50 kDa contributes to the drinking response of beer-taste beverages and does not produce aroma components having a low threshold value by reaction with sugar like amino acids. NS.

The beer-taste beverage of the present invention contains a protein having a molecular weight of 35 to 50 kDa.

The mass ratio of the protein content having a molecular weight of 35 to 50 kDa (protein with a molecular weight of 35 to 50 kDa / total protein) to the total protein content in the beer-taste beverage of the present invention has a drinkable response and flavor stability. From an excellent viewpoint, it is 0.005 or more, preferably 0.008 or more, more preferably 0.012 or more, still more preferably 0.014 or more, and the upper limit is not particularly limited, but is, for example, 0.1 or less. , 0.05 or less, 0.03 or less, etc., and may be in the range of any combination of these.

As a means for adjusting the mass ratio of the protein having a molecular weight of 35 to 50 kDa within the above range, the addition of the protein having a molecular weight of 35 to 50 kDa, the use of a raw material containing a large content of the 35 to 50 kDa protein, and the 35 to 50 kDa protein depending on the fermentation conditions. For example, controlling the content of the protein.

When adding a protein having a molecular weight of 35 to 50 kDa, it is preferable to add a protein derived from wheat. Examples of such wheat include barley, wheat, rye, oats, oats, and barley, and barley is preferable. Further, either germinated wheat or ungerminated wheat may be used, but germinated malt is preferable. These may be contained alone or in combination of two or more kinds.

When the content of 35 to 50 kDa protein is controlled by the fermentation conditions, it can also be controlled by controlling the fermentation temperature. For example, by lowering the temperature, the loss of 35 to 50 kDa protein due to foam separation by fermentation can be reduced, and as a result, more 35 to 50 kDa protein can be contained in the beer.

In the present specification, the 35 to 50 kDa protein is quantified by the Lowry method. The specific measurement method is shown below.

1.35 to 50 kDa Protein Purification 1) Protein Concentration with Ammonium Sulfate Add 3,900 g of ammonium sulfate (60% saturated ammonium sulfate) to 10 L of beer, stir for 3 hours with a stirrer, and centrifuge (12,000 g, 4 ° C.). 1 hour) to obtain a precipitate. Suspend the resulting precipitate in as little 20 mM phosphate buffer (pH 9.0) as possible and add 20 mM phosphate buffer (pH 9.0) until turbidity is removed. Then, ultrafiltration is performed using Amicon Ultra-15 (10 KDa cut off, Merck, UFC901024) (centrifugal at 2,800 g at 4 ° C until the final volume is about 1 ml). Then, about 10 ml of 20 mM phosphate buffer (pH 9.0) is added, and centrifugation is performed again under the same conditions to remove unnecessary ammonium sulfate. The supernatant thus obtained is used as a beer protein concentrated fraction in the next operation.

2) Fractionation of beer protein concentrated fraction by cation resin
SP Sepharose Fast Flow (manufactured by GE Healthcare Life Sciences) 100 ml is packed in an econocolumn, 300 ml of water is poured, and then 300 ml of 20 mM acetate buffer (pH 4.5) is poured for equilibration.
Prepare a beaker and add 20 mM acetate buffer (pH 4.5) to the beer protein concentrated fraction obtained in 1) to prepare 400 ml. Add 100 ml of SP Sepharose equilibrated with acetic acid buffer (pH 4.5) to the solution, and perform batch adsorption over 3 hours while stirring with a spatula or the like about every 10 minutes. After that, the contents of the beaker are packed in a column and the passing fractions are collected (FT). Then, 300 ml of 20 mM acetate buffer (pH 4.5) is loaded (A). Below, 20 mM acetate buffer (pH 4.5) containing 0.1 M NaCl 300 ml (B), 20 mM acetate buffer (pH 4.5) containing 0.2 M NaCl 300 ml (C), 20 mM acetate buffer containing 0.3 M NaCl (pH 4.5). ) Load 300 ml (D), 300 ml (E) of 20 mM acetate buffer (pH 4.5) containing 0.5 M NaCl. The obtained fractions are analyzed by SDS-PAGE and fractions containing 35-50 kDa protein are collected. Fractions containing these 35 to 50 kDa proteins are used as beer protein cation exchange resin binding fractions in the next operation.

3) Concentration of beer protein cation exchange resin bound fraction with ammonium sulfate Add 430 g of ammonium sulfate to 1 L of beer protein cation exchange resin bound fraction with stirring in a beaker. After stirring for 3 hours, the contents of the beaker are transferred to a centrifuge tube and centrifuged (12,000 g, 4 ° C., 3 hours) to obtain a precipitate.
Suspend the precipitate in as little 20 mM acetate buffer (pH 4.5) as possible and add 20 mM acetate buffer (pH 4.5) until turbidity is removed. Then, ultrafiltration is performed using Amicon Ultra-15 (10 KDa cut off, Merck, UFC901024) (centrifugal at 2,800 g at 4 ° C until the final volume is about 1 ml). Then, about 10 ml of 20 mM acetate buffer (pH 4.5) is added, and centrifugation is performed again under the same conditions to remove unnecessary ammonium sulfate. The supernatant thus obtained is used as a beer protein cation exchange resin bound fraction concentrate in the next operation.

4) Fraction of beer protein cation exchange resin-bonded fraction concentrate by anion resin
Q Sepharose Fast Flow (manufactured by GE Healthcare Life Sciences) 100 ml is packed in an econocolumn, 300 ml of water is poured, and then 300 ml of 20 mM phosphate buffer (pH 9.0) is poured for equilibration.
Prepare a beaker and add 20 mM phosphate buffer (pH 9.0) to the beer protein cation exchange resin-bonded fraction concentrate obtained in 3) to adjust to 400 ml. Add 100 ml of Q Sepharose equilibrated with 20 mM phosphate buffer (pH 9.0) to the solution, and perform batch adsorption over 3 hours while stirring with a spatula or the like about every 10 minutes. After that, the contents of the beaker are packed in a column and the passing fractions are collected (FT). Then load 500 ml of 20 mM phosphate buffer (pH 9.0) (A). 20 mM phosphate buffer (pH 9.0) containing 0.1 M NaCl, 300 ml (B), 20 mM phosphate buffer (pH 9.0) containing 0.2 M NaCl 300 ml (C), 20 mM phosphate buffer containing 0.3 M NaCl (pH 9.0). Load 300 ml (D) of pH 9.0), 300 ml (E) of 20 mM phosphate buffer (pH 9.0) containing 0.5 M NaCl. The obtained fractions FT and A to E are analyzed by SDS-PAGE, respectively, and the fractions containing 35 to 50 kDa protein are collected. Fractions containing these 35 to 50 kDa proteins are used as 35 to 50 kDa protein ion exchange resin-bound fractions in the next operation.
For (A), (B), (C), (D), and (E), 0.5 M disodium phosphate is being added at a ratio of 15 ml to 500 ml of liquid immediately after column elution. Reconcile. The operation of 4) is carried out within one day in order to minimize the change in protein due to alkali.

5) Concentration of 35 to 50 kDa protein ion exchange resin-bound fraction with ammonium sulfate Add 430 g of ammonium sulfate to 1 L of the 35 to 50 kDa protein ion exchange resin-bound fraction with stirring in a beaker. After stirring for 3 hours, the contents of the beaker are transferred to a centrifuge tube and centrifuged (12,000 g, 4 ° C., 3 hours) to obtain a precipitate.
Suspend the precipitate in as little 20 mM acetate buffer (pH 4.5) as possible and add 20 mM acetate buffer (pH 4.5) until turbidity is removed. Then, ultrafiltration is performed using Amicon Ultra-15 (10 KDa cut off, Merck, UFC901024) (centrifugal at 2,800 g at 4 ° C until the final volume is about 1 ml). Then, about 10 ml of 20 mM acetate buffer (pH 4.5) is added, and centrifugation is performed again under the same conditions to remove unnecessary ammonium sulfate. The supernatant thus obtained is used for analysis as a purified product of 35 to 50 kDa protein.

2. Protein quantification by the Lowry method Concentration occurs when preparing a 35-50 kDa protein purified product. That is, the volume of the 35 to 50 kDa protein purified product obtained from the beer-taste beverage is smaller than the volume of the beer-taste beverage. Therefore, the protein content of the 35-50 kDa protein purified product of the beer-taste beverage in the present invention is the protein content measured using the 35-50 kDa protein purified product prepared from the beer-taste beverage. Concentration rate when the 35-50 kDa protein purified product was prepared (that is, the volume of the beer-taste beverage used in the preparation was divided by the volume of the 35-50 kDa protein purified product obtained in the preparation. It is calculated by dividing by the obtained ratio). The protein quantification was performed by the Lowry method using a commercially available kit (DC protein assay, manufactured by Bio-Rad). First, the concentration of the fractionated solution was adjusted so as to be within an appropriate range. To 5 μL of the sample whose concentration was adjusted, 50 μL of solution A was added and stirred, and then 400 μL of solution B was added and stirred. After the color reaction was carried out at room temperature for 15 minutes, 350 μL was transferred to a 96-well plate and the absorbance at 750 nm was measured. The peptide concentration (mg / mL) was calculated based on the obtained absorbance and the calibration curve prepared in advance. The calibration curve was prepared using BSA (bovine serum albumin). Based on the calibration curve, the content of 35 to 50 kDa protein in the purified 35 to 50 kDa protein is calculated.

Further, in the present specification, the total protein content is calculated by multiplying the total nitrogen content by a protein conversion coefficient of 6.25. Here, the total amount of nitrogen is measured by the Kjeldahl method. Nitrogen compounds in beer-taste beverages are not necessarily proteins, but may be amino acids, amides, purine bases, creatines, etc., but total nitrogen is regarded as derived from proteins and converted.

The method for producing a beer-taste beverage of the present invention is not particularly limited, and examples thereof include a production method having a step of adding 35 to 50 kDa protein. More specifically, an embodiment in which a wheat-derived protein having a molecular weight of 35 to 50 kDa is added is mentioned, and the production method in this embodiment is illustrated below. In addition, as another embodiment, the conditions in the manufacturing process are conventional, such as using a high nitrogen malt raw material produced in North America having a high content of 35 to 50 kDa protein, or increasing the amount of 35 to 50 kDa protein depending on the fermentation conditions. It may be increased by setting a condition for producing more wheat-derived protein having a molecular weight of 35 to 50 kDa than the condition.

The production method of the embodiment in which the wheat-derived protein having a molecular weight of 35 to 50 kDa is added (the production method of this embodiment) is the same as the production method of a general beer-taste beverage except that the step of adding the 35 to 50 kDa protein is included. The production mode of the beer-taste beverage will be illustrated below. There are two types of beer-taste beverages, one in which malt is used as a raw material and the other in which malt is not used as a raw material, and the beer-taste beverage can be produced as follows.

Alcohol-containing beer-taste beverages produced using malt as a raw material include, first of all, in addition to wheat such as malt, other grains such as starch, starch, sugar, bitterness, or coloring agent as needed. If necessary, an enzyme such as amylase is added to the mixture containing water to gelatinize and saccharify, and the mixture is filtered to obtain a saccharified solution. If necessary, add hops and bitterness to the saccharified solution and boil, and remove solids such as coagulated protein in a clarification tank. As an alternative to this saccharified solution, hops may be added to a malt extract mixed with warm water and boiled. Hops may be mixed at any stage from the start of boiling to the end of boiling. As the conditions in the saccharification step, the boiling step, the solid content removing step and the like, known conditions may be used. As the conditions in the fermentation / liquor storage process and the like, known conditions may be used. The obtained fermentation broth is filtered, and carbon dioxide gas is added to the obtained filtrate. After that, it is filled in a container and sterilized to obtain a desired beer-taste beverage. In addition, as an alcohol component, spirits derived from grains may be further added. Spirits means alcoholic beverages obtained by fermenting grains such as wheat, rice, buckwheat, and corn with yeast and then further distilling them. Wheat is preferred as the grain that is the raw material for spirits. The step of adding the 35 to 50 kDa protein in each of the steps may be performed in any step up to filling.

Alcohol-containing beer-taste beverages produced without using malt as a raw material are a mixture of liquid sugar containing a carbon source, nitrogen source as an amino acid-containing material other than wheat or malt, hops, pigments, etc. together with warm water. Then, use a liquid sugar solution. The liquid sugar solution is boiled. When hops are used as a raw material, the hops may be mixed with the liquid sugar solution during boiling, not before the start of boiling. As an alternative to this saccharified solution, hops may be added to an extract using a raw material other than malt with warm water and boiled. Hops may be mixed at any stage from the start of boiling to the end of boiling. As the conditions in the fermentation / liquor storage process and the like, known conditions may be used. The obtained fermentation broth is filtered, and carbon dioxide gas is added to the obtained filtrate. After that, it is filled in a container and sterilized to obtain a desired beer-taste beverage. In addition, as an alcohol component, spirits derived from grains may be further added. Spirits means alcoholic beverages obtained by fermenting grains such as wheat, rice, buckwheat, and corn with yeast and then further distilling them. Wheat is preferred as the grain that is the raw material for spirits. The step of adding the 35 to 50 kDa protein in each of the steps may be performed in any step up to filling.

The non-fermented and alcohol-containing beer-taste beverage may or may not use malt, and the alcohol content of the final product may be adjusted by adding alcohol for raw materials. The alcohol for raw materials may be added in any step from the saccharification step to the filling step. In addition, as an alcohol component, spirits derived from grains may be further added. Spirits means alcoholic beverages obtained by fermenting grains such as wheat, rice, buckwheat, and corn with yeast and then further distilling them. Wheat is preferred as the grain that is the raw material for spirits. The step of adding the 35 to 50 kDa protein in each of the steps may be performed in any step up to filling.

For non-alcoholic beer-taste beverages produced using malt as a raw material, first, in addition to wheat such as malt, other grains, starch, saccharides, bitterness, or coloring agents and water are added as needed. If necessary, an enzyme such as amylase is added to the containing mixture to gelatinize and saccharify, and the mixture is filtered to obtain a saccharified solution. If necessary, add hops and bitterness to the saccharified solution and boil, and remove solids such as coagulated protein in a clarification tank. As an alternative to this saccharified solution, hops may be added to a malt extract mixed with warm water and boiled. Hops may be mixed at any stage from the start of boiling to the end of boiling. As the conditions in the saccharification step, the boiling step, the solid content removing step and the like, known conditions may be used. After boiling, the obtained wort is filtered, and carbon dioxide gas is added to the obtained filtrate. Then, it is filled in a container and sterilized to obtain a desired non-alcoholic beer-taste beverage. The step of adding the 35 to 50 kDa protein in each of the steps may be performed in any step up to filling.

When producing a non-alcoholic beer-taste beverage that does not use malt as a raw material, first, liquid sugar containing a carbon source, nitrogen source as an amino acid-containing material other than wheat or malt, hops, pigments, etc. are mixed with warm water. Then, use a liquid sugar solution. The liquid sugar solution is boiled. When hops are used as a raw material, the hops may be mixed with the liquid sugar solution during boiling, not before the start of boiling. Carbon dioxide gas is added to the liquid sugar solution after boiling. Then, it is filled in a container and sterilized to obtain a desired non-alcoholic beer-taste beverage. The step of adding the 35 to 50 kDa protein in each of the steps may be performed in any step up to filling.

In the production method of this embodiment, an aliphatic alcohol may be added from the viewpoint of imparting a feeling of alcohol. The aliphatic alcohol is not particularly limited as long as it is known, but an aliphatic alcohol having 4 to 5 carbon atoms is preferable. In the production method of this embodiment, preferred aliphatic alcohols include 2-methyl-1-propanol and 1-butanol as those having 4 carbon atoms, and 3-methyl-1-butanol as those having 5 carbon atoms. Examples thereof include 1-pentanol and 2-pentanol. These can be used in one type or a combination of two or more types. The content of the aliphatic alcohol having 4 to 5 carbon atoms is preferably 0.0002 to 0.0007% by mass, and more preferably 0.0003 to 0.0006% by mass. In the present specification, the content of the aliphatic alcohol can be measured by using a headspace gas chromatograph method.

(Acidulant)
As the acidulant used in the production method of this embodiment, it is preferable to use one or more acids selected from the group consisting of citric acid, lactic acid, phosphoric acid, and malic acid. Further, in the production method of this embodiment, succinic acid, tartaric acid, fumaric acid, glacial acetic acid and the like can also be used as the acid other than the acid. These can be used without limitation as long as they are approved to be added to foods. In the production method of this embodiment, it is preferable to use a combination of lactic acid from the viewpoint of appropriately imparting a mild acidity and phosphoric acid from the viewpoint of appropriately imparting a slightly irritating acidity.

The content of the acidulant in the beer-taste beverage obtained by the production method of this embodiment is preferably 200 ppm or more, more preferably 550 ppm or more, still more preferably 700 ppm or more, in terms of citric acid, from the viewpoint of imparting a beer-taste feeling. Further, from the viewpoint of acidity, 15000 ppm or less is preferable, 5500 ppm or less is more preferable, and 2000 ppm or less is further preferable. Therefore, in this embodiment, the content of the acidulant may be in a preferable range of 200 ppm to 15000 ppm, preferably 550 ppm to 5500 ppm, more preferably 700 ppm to 1500 ppm in terms of citric acid. In the present specification, the citric acid conversion amount is an amount converted from the acidity of each acidulant based on the acidity of citric acid. For example, the citric acid conversion amount corresponding to 100 ppm of lactic acid is used. The citric acid conversion amount corresponding to 120 ppm and 100 ppm of phosphoric acid is converted as 200 ppm, and the citric acid conversion amount corresponding to 100 ppm of malic acid is converted as 125 ppm.

The content of the acidulant in the beer-taste beverage refers to the one calculated by analysis by high performance liquid chromatography (HPLC) or the like.

(hop)
In the production method of this embodiment, hops can be used as a part of the raw material. Hops are desirable as part of the raw material, as the flavor tends to resemble beer. When hops are used, ordinary pellet hops, powdered hops, and hop extracts used in the production of beer and the like can be appropriately selected and used according to a desired flavor. Further, processed hop products such as isometric hops and reduced hops may be used. These are included in the hops used in the production method of this embodiment. The amount of hops added is not particularly limited, but is typically about 0.0001 to 1% by mass with respect to the total amount of the beverage.

(Other raw materials)
In the production method of this embodiment, other raw materials may be optionally used. For example, sweeteners (including high-sweetness sweeteners), bitterness, flavors, yeast extracts, colorants such as caramel pigments, plant-extracted saponin-based substances such as soybean saponin and kiraya saponin, plant proteins such as corn and soybean, and Peptide-containing substances, animal proteins such as milk syrup, seasonings such as dietary fiber and amino acids, and antioxidants such as ascorbic acid can be used as needed as long as they do not interfere with the effects of this embodiment.

The pH of the beer-taste beverage obtained by the production method of this embodiment is preferably 3.0 to 5.0, more preferably 3.0 to 4.5, from the viewpoint of improving the flavor of the beverage. More preferably, it is 3.0 to 4.0.

(Beverage in a container)
The beer-taste beverage obtained by the production method of this embodiment can be packed in a container. The form of the container is not limited at all, and it can be filled in a sealed container such as a bottle, a can, a barrel, or a PET bottle to obtain a beverage in a container.

Hereinafter, the present invention will be specifically described with reference to Examples, but the present invention is not limited to the following Examples.

Preparation Example 1: Preparation of 35 to 50 kDa protein A 35 to 50 kDa protein was prepared according to the description in the above section “Purification of 35 to 50 kDa protein”.

Preparation Example 2: Preparation of amino acid mixture
In Eur Food Res Technol (2010) 230: 665-673, various amino acid reagents were mixed so as to have the same amino acid ratio as the amino acid sequence of Protein Z shown in Fig. 4, and an amino acid mixed solution was prepared.

Preparation of beer-taste beverage Reference Example 1, Examples 1-5
30 kg of malt was crushed to an appropriate particle size, placed in a charging tank, and then 120 L of warm water was added to make a mash at about 50 ° C. A part was heated to 100 ° C. and boiled, and the rest was saccharified. After the saccharified mash was heated to 78 ° C., it was transferred to a wort filtration tank and filtered to obtain a filtrate. Hops were added to the filtrate and then boiled for 80 minutes to obtain prepared wort. Hops were added as needed even after the start of boiling. Brewer's yeast was added to the prepared wort and fermented at about 15 ° C. for about 15 days to obtain a stored beer. Next, the beer-taste beer of Reference Example 1 was produced by filtering the stored beer without heat treatment to remove yeast. When the beer-taste beverage of Reference Example 1 was analyzed according to the above measurement method, the content of 35 to 50 kDa protein was 12 ppm, and the content of total protein was 3000 ppm. The 35 to 50 kDa protein obtained in Preparation Example 1 was added to this beer-taste beverage so as to have the concentration shown in Table 1 to obtain the beer-taste beverage of Examples 1 to 5.

Comparative Examples 1 to 3
A beer-taste beverage was prepared in the same manner as in Examples 1 to 5 except that the amino acid mixture obtained in Preparation Example 2 was added in place of the 35 to 50 kDa protein of Preparation Example 1 so as to have the concentration shown in Table 2. Obtained.

Evaluation of flavor The flavors of Examples 1 to 5 and Comparative Examples 1 to 3 were evaluated by a sensory test. Five well-trained sensory evaluators evaluated "richness" and "freshness" on a scale of 5 points. The richness was evaluated using the one stored at 4 ° C. for 3 weeks after production. The evaluation of the freshness was carried out using a product stored at 28 ° C. for 3 weeks after production.

Regarding the richness, the average score of the evaluation points is 5 points for "very feel", 4 points for "feel", 3 points for "slightly feel", 2 points for "slightly feel", and 1 point for "not feel". Was calculated and evaluated according to the following criteria according to the average score. Regarding the freshness (that is, flavor stability), "very good" is 5 points, "good" is 4 points, "somewhat good" is 3 points, "somewhat bad" is 2 points, and "bad" is 1 point. The average score of the evaluation points was calculated, and the evaluation was performed according to the following criteria according to the average score. The results are shown in Tables 1 and 2. Reference Example 1 was evaluated as 1 point for richness, and Reference Example 1 was evaluated as 5 points for freshness.

<Criteria for evaluation points>
X: Average value 1.0 or more and less than 2.0 Δ: Average value 2.0 or more and less than 3.0 ○: Average value 3.0 or more and less than 4.0 ◎: Average value 4.0 or more and less than 5.0. 0 or less

As can be seen from Table 1, the beer-taste beverages of Examples 1 to 5 in which the mass ratio of the protein content having a molecular weight of 35 to 50 kDa to the total protein content is 0.005 or more and 0.1 or less are rich. It had excellent flavor stability. On the other hand, as can be seen from Table 2, in Comparative Examples 1 to 3 to which the amino acid mixed solution was added, although the richness could be imparted, there was a problem in flavor stability.

According to the present invention, it is possible to provide a beer-taste beverage having excellent flavor stability while imparting richness.

Claims (2)

A beer-taste beverage in which the mass ratio of the protein content having a molecular weight of 35 to 50 kDa to the total protein content is 0.005 or more and 0.1 or less. The beer-taste beverage according to claim 1, wherein the mass ratio is 0.008 or more and 0.05 or less.