TW202214115A - Non-alcoholic beer-flavored beverage - Google Patents

Abstract

A non-alcohol beer-flavored beverage containing malt as a raw material, the beverage comprising one or more diketopiperazines selected from the group consisting of cycloleucyl proline, cyclovalyl proline, and cycloisoleucyl proline, wherein the concentration of the most abundant one of the diketopiperazines is at least 0.4 ppm.

Description

Non-alcoholic beer-flavored beverages

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

With the diversification of consumers' tastes in recent years, the development of a non-alcoholic beer-flavored beverage having various flavor characteristics is desired.

Patent Document 1 discloses that a peptide having a specific molecular weight is contained in order to improve the flavor of a beer-flavored beverage. [Prior Art Literature] [Patent Literature]

[Patent Document 1] Japanese Patent Laid-Open No. 2016-149975

[Problems to be Solved by the Invention]

Patent Document 1 describes a beverage, which is used as an evaluation index for the beverage, using such indicators as beer-like flavor, smooth taste flow, and gritty feeling remaining in the mouth. , the sensory evaluation score based on these indicators is higher.

Here, as a taste immediately after drinking a beer-flavored beverage, the five basic tastes, namely, sweet, salty, sour, bitter, and umami, cannot be expressed as the intensity, diffusion, and richness of the taste. Characteristics of a balance of intensity, flavor persistence or flavor intensity are preferred. Such a feature is called "swelling degree" in this specification. From the viewpoint of the swelling degree, the beverage described in Patent Document 1 can be said to have room for improvement, and a method for enhancing the swelling degree is desired.

The purpose of the present invention is to provide a beverage with enhanced swelling. In particular, the object of the invention is to provide a beverage with enhanced swelling in a non-alcoholic beer-flavored beverage containing malt as a raw material. Since the non-alcoholic beer-flavored beverage containing malt in the raw material tends to lack the swelling degree, a beverage with an enhanced swelling degree is particularly desired. [Means of Solving Problems]

That is, the present invention relates to the following non-alcoholic beer-flavored beverages. [1] A non-alcoholic beer-flavored beverage, which is a non-alcoholic beer-flavored beverage containing malt in the raw material, and comprises a non-alcoholic beer-flavored beverage selected from the group consisting of cyclomethicone, cyclovalinylproline, and cycloisolem At least one diketopiperazine in the group consisting of acylproline, and among the above-mentioned diketopiperazine, the concentration of the most contained diketopiperazine is 0.4 ppm or more. [2] The non-alcohol beer-flavored beverage according to the above [1], wherein the diketopiperazine most contained in the diketopiperazine has a concentration of 10 ppm or less. [3] The non-alcoholic beer-flavored beverage according to the above [1] or [2], further comprising a protein having a molecular weight of 35 to 50 kDa, and the concentration of the protein is 5 ppm or more. [4] The non-alcoholic beer-flavored beverage according to the above [3], wherein the concentration of the protein is 30 ppm or less. [Effect of invention]

According to the present invention, it is possible to provide a beverage with enhanced swelling among non-alcoholic beer-flavored beverages containing malt as a raw material.

The non-alcohol beer-flavored beverage system raw material of the present invention contains malt. The raw materials mentioned here mean grain raw materials and sugars other than water and hops. As a raw material, other than malt, rice, corn, sorghum, potato, starch, and wheat other than malt may be included. Components such as acidulants, sweeteners, bitternesses, seasonings, and spices that can be added in trace amounts are not included in the raw materials. The weight of the extract in the non-alcoholic beer-flavored beverage of the present invention is not particularly limited, but is preferably 0.01 to 20.0% by weight.

Non-alcoholic beer-flavored beverages refer to beer-flavored beverages with an alcohol content of less than 1%, preferably substantially free of alcohol. Also, the alcohol content may be 0%. The beer-flavored beverage means a beer-flavored carbonated beverage. Here, a beverage in a form that does not substantially contain alcohol is not a beverage that contains an undetectable amount of alcohol in a very small amount. Beverages whose alcohol content is rounded to the nearest 0.0% are also included in non-alcoholic beer-flavored beverages. Examples of non-alcoholic beer-flavored beverages of the present invention include non-alcoholic beer-flavored beverages, beer-flavored soft drinks, and the like. In addition, the "alcohol content (alcohol content)" here means the content of ethanol, and does not include the content of aliphatic alcohols other than ethanol.

The alcohol content of the non-alcoholic beer-flavored beverage according to the present invention means the content (v/v%) of the alcohol component in the beverage, and can be measured by any known method, but can be measured by, for example, a vibrating density meter. Specifically, it is possible to prepare a sample for removing carbonic acid gas from beverages by filtration or ultrasonic waves, and subject the sample to direct-fire distillation to measure the density of the obtained distillate at 15°C, using the analytical method specified by the National Taxation Bureau (flat). 19. Order No. 6 of the National Taxation Bureau (revised on June 22, 2019) in the attached table "Table 2 Alcohol Composition and Density (15°C) and Specific Gravity (15/15°C) Conversion Table" to convert and calculate. When the alcohol content is less than 1.0%, a commercially available alcohol measuring device or gas chromatography can also be used.

The non-alcoholic beer-flavored beverage of the present invention comprises at least one diketopiperazine selected from the group consisting of cycloleukinylproline, cyclovalinylproline, and cycloleukinylproline . Diketopiperazines are cyclic dipeptides bonded to two amino acids. Cyclolemacylproline is a bond between leucine and proline, and may be referred to as Cyclo (Leu-Pro) in this specification. Cyclovalinylproline is a bond between valine and proline, and may be referred to as Cyclo (Val-Pro) in this specification. Cycloproline is a bond between isoleucine and proline, and may be referred to as Cyclo (Ile-Pro) in this specification.

In the non-alcohol beer-flavored beverage of the present invention, among the above-mentioned diketopiperazine, the concentration of the diketopiperazine contained the most is 0.4 ppm or more. If the concentration of these diketopiperazine is 0.4 ppm or more, the swelling degree in a non-alcoholic beer-flavored beverage can be enhanced. It is a fact that the inventors of the present invention have found that it has not been known so far that the specific diketopiperazine contained in a specific concentration or more is related to the swelling degree in a non-alcohol beer-flavored beverage.

In the non-alcoholic beer-flavored beverage of the present invention, among the diketopiperazine, the concentration of the most contained diketopiperazine is preferably 10 ppm or less.

In addition, the non-alcoholic beer-flavored beverage of the present invention further contains a protein with a molecular weight of 35 to 50 kDa, and the concentration of the protein is preferably 5 ppm or more.

The protein with a molecular weight of 35 to 50 kDa is a protein with a molecular weight of 35 to 50 kDa when a non-alcoholic beer flavored beverage is subjected to SDS-PAGE electrophoresis. Before supplying the non-alcoholic beer-flavored beverage to SDS-PAGE electrophoresis, for example, as a pretreatment, the non-alcohol beer-flavored beverage may be subjected to extra-limited filtration using a 30 kDa cut off membrane. The above-mentioned protein is preferably a protein with a molecular weight of 35-45 kDa, more preferably a protein of about 40 kDa. In this specification, proteins with a molecular weight of 35 to 50 kDa are also referred to as 40 kDa proteins.

The 40kDa protein is preferably a protein derived from cereals. It is preferable that the above-mentioned cereals are at least one selected from the group consisting of barley, wheat, corn, rice, and soybeans. In addition, when the grain is wheat, a well-known wheat-derived protein used for the production of a non-alcoholic beer-flavored beverage can be contained. Examples of such wheat include barley, wheat, rye, black wheat, barley, oats, and the like, and barley is preferred. In addition, it may be either malted wheat or unmalted wheat, but it is preferably malted wheat malt. These may be contained individually or in combination of 2 or more types.

Further, as the 40 kDa protein, Serpin Z4 (alias: BSZ4, HorvuZ4, Major endosperm albumin or Protein Z) derived from barley (scientific name: Hordeum vulgare) and Serpin Z7 (alias: BSZ7 or HorvuZ7) derived from barley are preferable. In addition, among the above-mentioned proteins, amino acids which are part of the amino acid sequence may also have amino acid sequences deleted, substituted, inserted and/or added.

By further containing 40 kDa protein in addition to the diketopiperazine, the swelling degree in the non-alcoholic beer-flavored beverage can be further enhanced. Furthermore, the concentration of the 40 kDa protein is preferably 30 ppm or less.

Moreover, when the diketopiperazine and the 40kDa protein are simultaneously contained, the swelling degree of the non-alcoholic beer flavor beverage can be effectively enhanced by the synergistic effect of the diketopiperazine and the 40kDa protein.

The following shows the production steps of general non-alcoholic beer-flavored beverages. A non-alcoholic beer-flavored beverage can be easily produced by the fermentation step without yeast.

When manufacturing a non-alcoholic beer-flavored beverage using malt as a raw material, first, in addition to wheat such as malt, other raw materials and water such as grains, starch, sugar, bitterness, coloring, etc. are included as necessary. In the mixture, enzymes such as amylase are added as necessary to carry out gelatinization and saccharification, and then filtered as a saccharification solution. If necessary, add hops, bitters, etc. to the saccharification solution, boil it, and remove solids such as coagulated proteins in a clarification tank. As an alternative to this saccharification solution, hops may be added to the malt extract with warm water added and boiled. Hops can also be blended at any stage from the start of boiling until the end of boiling. The conditions in the saccharification step, boiling step, solid content removal step, etc. may be well-known conditions. After boiling, the obtained wort was filtered, and carbon dioxide gas was added to the obtained filtrate. After that, through the sterilization step of filling the container, the intended non-alcoholic beer-flavored beverage is obtained.

In the non-alcohol beer-flavored beverage according to the present invention, an aliphatic alcohol may be added from the viewpoint of imparting a alcoholic feel. The aliphatic alcohol is not particularly limited as long as it is a known one, but an aliphatic alcohol having 4 to 5 carbon atoms is preferable. In the present invention, as preferable aliphatic alcohols, those having 4 carbon atoms include 2-methyl-1-propanol, 1-butanol, and the like, and those having 5 carbon atoms include 3-methyl- 1-butanol, 1-pentanol, 2-pentanol, etc. These can be used alone or in combination of two or more. The content of the aliphatic alcohol having 4 to 5 carbon atoms is preferably 0.0002 to 0.0007% by weight, more preferably 0.0003 to 0.0006% by weight. In the present specification, the content of the aliphatic alcohol can be measured using headspace gas chromatography.

The non-alcoholic beer-flavored beverage related to the present invention caters to the low-calorie preference in recent years, and is expected to be low-calorie. Therefore, the calorie count of the non-alcoholic beer-flavored beverage related to the present invention is preferably less than 5kcal/100mL, more preferably less than 4kcal/100mL, more preferably less than 3kcal/100mL.

The number of calories contained in the non-alcoholic beer-flavored beverage according to the present invention is basically calculated based on the "Analysis method for nutritional components, etc., based on the nutrition representation standard" published in the Related Health Promotion Act. That is, in principle, it can be used as a calorie conversion factor (protein: 4kcal/g, lipid: 9kcal/g, carbohydrate: 4kcal/g, dietary fiber: 2kcal/g) by multiplying the quantitative amount of each nutritional component by the calorie conversion factor of each component. , alcohol: 7kcal/g, organic acid: 3kcal/g) sum to calculate. For details, please refer to "Analysis methods, etc., of nutrient components, etc. on the basis of nutritional representation."

The specific method for measuring the amount of each nutrient contained in the non-alcoholic beer-flavored beverage according to the present invention may be in accordance with the various analytical methods described in the Health Promotion Act "Analysis of Nutrients, etc. Regarding Nutritional Expression Standards". In addition, if you contact the Japan Food Analysis Center, you can know such calories and/or the amount of each nutrient.

The saccharide contained in the non-alcohol beer-flavored beverage according to the present invention means the saccharide based on the nutrition representation standard for foods (Ministry of Health, Labour and Welfare Notice No. 176 in 2015). Specifically, a carbohydrate means a protein, lipid, dietary fiber, ash, alcohol content, and water that are removed from food. Also, the amount of sugar in a food can be calculated by deducting the amounts of protein, lipid, dietary fiber, ash, and moisture from the weight of the food. At this time, the amounts of protein, lipid, dietary fiber, ash, and moisture were measured by the methods disclosed in the Nutritional Standard. Specifically, the amount of protein was measured by nitrogen quantitative conversion method, the amount of lipid was measured by ether extraction method, chloroform-methanol mixed liquid extraction method, Gerber method, acid decomposition method or Reese-Gottlieb method, dietary fiber The amount of ash is determined by high-speed liquid chromatography or Prosky method, the amount of ash is determined by magnesium acetate adding ashing method, direct ashing method or sulfuric acid adding ashing method, and the amount of moisture is determined by Karl-Fischer method, drying aid method. Dosage method, reduced pressure superheat drying method, atmospheric pressure heating drying method or plastic film method to measure.

The non-alcoholic beer-flavored beverage related to the present invention caters to the low-sugar content preference in recent years, and can also be low-sugar content. Therefore, the saccharide content of the non-alcoholic beer-flavored beverage related to the present invention may be less than 2.5 g/100 mL or less than 0.5 g/100 mL. In addition, the lower limit is not particularly set, but is usually about 0.1 g/100 mL, for example, 0.15 g/100 mL or more or 0.2 g/100 mL or more.

The non-alcoholic beer-flavored beverage related to the present invention may also contain a sour taste. As an acidulant, it is preferable to use 1 or more types of acid chosen from the group which consists of citric acid, lactic acid, phosphoric acid, and malic acid. Furthermore, in the present invention, succinic acid, tartaric acid, butenedioic acid, glacial acetic acid, and the like can also be used as acids other than the above-mentioned acids. These can be used without limitation as long as they are approved to be added to foods. In the present invention, lactic acid and phosphoric acid are preferably used in combination from the viewpoint of appropriately imparting a mild acidity and a viewpoint of appropriately imparting a slightly irritating acidity.

In the non-alcoholic beer-flavored beverage related to the present invention, the content of the sour flavor is converted to citric acid, and from the viewpoint of imparting a beer flavor, it is preferably 200 ppm or more, more preferably 550 ppm or more, and more preferably 700 ppm or more. Furthermore, from the viewpoint of acidity, it is preferably 15,000 ppm or less, more preferably 5,500 ppm or less, and more preferably 2,000 ppm or less. Therefore, in the present invention, the content of the sour taste is converted to citric acid, and there are suitable ranges such as 200ppm to 15000ppm, preferably 550ppm to 5500ppm, and more preferably 700ppm to 1500ppm. In addition, in this specification, the citric acid conversion amount means the amount converted from the acidity degree of each acidulant based on the acidity degree of citric acid. The citric acid equivalent of 100 ppm of phosphoric acid was converted as 200 ppm, and the citric acid equivalent of 100 ppm of malic acid was converted as 125 ppm.

The content of the acidulant in the non-alcohol beer-flavored beverage means what is calculated by analysis such as high-speed liquid chromatography (HPLC).

In the non-alcoholic beer-flavored beverage according to the present invention, hops can be used as part of the raw material. When hops are used, general flake hops, powdered hops, and hop extracts used in the production of beer and the like can be appropriately selected and used according to the desired flavor. In addition, processed hops such as dissimilated hops and reduced hops can also be used. These are included in the hops used in the non-alcoholic beer-flavored beverage related to the present invention. In addition, the addition amount of hops is not particularly limited, but is typically about 0.0001 to 1% by weight with respect to the total amount of the beverage.

In the non-alcoholic beer-flavored beverage related to the present invention, other raw materials may be used as necessary within the range that does not hinder the effects of the present invention. Coloring materials such as sweeteners (including high-intensity sweeteners), bitters, flavors, yeast extracts, caramel colors, and soybean saponins can be used as necessary within the range that does not hinder the effects of the present invention. Or plant extract saponins such as saponins, plant proteins such as corn or soybeans and substances containing peptides, bovine serum albumin and other protein-based substances, dietary fibers or amino acids and other seasonings, ascorbic acid, etc. of antioxidants.

The non-alcoholic beer-flavored beverage related to the present invention can be packaged as a container. There is no restriction on the form of the container, and it can be filled in sealed containers such as bottles, cans, barrels, or pet bottles, and can be used as beverages contained in containers.

The production method of the non-alcohol beer-flavored beverage of the present invention is not particularly limited, but a method of adding a specific amount of diketopiperazine to a non-alcohol beer-flavored beverage containing malt as a raw material is exemplified. Moreover, it is preferable to add 40kDa protein to the non-alcoholic beer-flavored beverage containing malt in the raw material. The preparation of the added diketopiperazine and the 40-kDa protein can be carried out by, for example, the procedure described in the examples below. Moreover, regarding the diketopiperazine and the 40 kDa protein, the content of these can also be increased by adjusting various conditions in the production process of the non-alcoholic beer-flavored beverage. [Example]

The present invention will be specifically described below with reference to examples, but the present invention is not limited to the following examples.

(Purification of Diketopiperazine) Purification of the diketopiperazine was carried out as follows. (1) Fractionation of HP20 of beer 60 L of beer was fractionated using 10 L of Diaion (registered trademark) HP20 (manufactured by Mitsubishi Chemical Co., Ltd.). The HP20 was washed three times with ethanol and then three times with 50% ethanol before use. The washed HP20 was packed in a column for a large number of fractionation and replaced with water. Mix the degassed 60L beer with the same amount of distilled water, use a medium pressure pump, and flow into the HP20 column. A solution that passed through the HP20 column was obtained as the pass fraction. Using a medium-pressure pump, 40 L of distilled water was poured into the solution to obtain an elution solution, which was used as a water elution fraction. Similarly, 40 L of water-containing ethanol (10% ethanol, 30% ethanol, and 70% ethanol) were each poured to obtain eluates, which were used as 10% ethanol eluted fractions, 30% ethanol eluted fractions, and 70% ethanol eluted fractions. The respective eluted fractions were refrigerated and stored as dried products using an evaporator and a freeze dryer.

(2) LH-20 Fractionation of 30% Ethanol Dissolution Fraction Among the HP20 fractions, 1.2 kg of Sephadex (registered trademark) LH-20 was used for fractionation of the 30% ethanol-eluted fraction. The LH-20 washed with ethanol was packed into a column for fractionation in large quantities, and replaced with water. 17.6 g of the 30% ethanol eluted fraction (87.9 g) obtained by fractional distillation with HP20 was dissolved in distilled water and used in an LH-20 column. Use a medium-pressure pump to flow in 13.5L of distilled water to obtain water-dissolved fractions -1~6. Next, 7 L of water-containing ethanol (35% ethanol, 70% ethanol and 100% ethanol) was poured into each to obtain elution liquids as 35% ethanol eluted fractions, 70% ethanol eluted fractions and 100% ethanol eluted fractions, respectively. The respective eluted fractions were refrigerated and stored as dried products using an evaporator and a freeze dryer.

(3) Separation of diketopiperazine Of the water-eluted fraction-3 (1.04 g) obtained by fractional distillation with LH-20, 86.7 mg was eluted with 10% ethanol using HPLC (COSMOSIL 5C18-PAQ, 20×250 mm). Next, the eluate was concentrated from 15 minutes to 20 minutes, and eluted with a mixture of a concentration gradient of ethanol-water (5:95→20:80) using HPLC (COSMOSIL 5C18-PAQ, 20×250 mm) to obtain Compound 1 (6.7 mg, tR=25 min). Similarly, the eluate was concentrated from 27.5 minutes to 40 minutes, and eluted with a concentration gradient mixture of ethanol-water (5:95→20:80) using HPLC (COSMOSIL 5C18-PAQ, 20×250 mm), Compound 2 (2.2 mg, tR=43 min) and compound 3 (2.6 mg, tR=46 min) were obtained. Compound 1 was identified as Cyclo (Pro-Val) based on the analysis of MS and NMR physical data and literature values. Compound 2 was identified as Cyclo (Ile-Pro) based on the analysis of MS and NMR physical data and literature values. Compound 3 was identified as Cyclo (Leu-Pro) based on the analysis of MS and NMR physical data and literature values. The reference cited is J. Agric. Food Chem. 2007, 55, 75-79. The analytical machine used is as described below. LC-MS; Q Exactive, manufactured by Thermo Fisher Scientific NMR; AVANCE400, manufactured by Bruker

(purification of 40kDa protein) Purification of the 40 kDa protein was performed from 1 L of commercial beer and as follows.

(1) Fractionation of cation exchange resin The cation exchange resin SP Sepharose 50mL was put into the empty column. Make the beer adsorb the resin. After that, the resin was converted into a column and washed with 20 mM sodium acetate buffer (pH 4.5). Next, it was eluted with 20 mM sodium acetate (pH 4.5) + 0.5 M-NaCl, and the fractions were collected. The resulting fractions were evaluated by SDS-PAGE, and the fractions contained in the 40 kDa protein were collected as cation exchange resin bound fractions.

(2) Filter outside the limit (buffer exchange) After washing with water, 10 mL of the cation exchange resin-bound fraction obtained in (1) was added to an out-of-limit filtration unit (Amicon Ultra-15 30K manufactured by Merck), and centrifuged at 3500 rpm to obtain an out-of-limit filtration concentrate.

(3) Ammonium sulfate fractionation 20 mM phosphate buffer (pH 7.0) + 2 M ammonium sulfate was added to the beaker, and the concentrated solution obtained in (2) was added dropwise and stirred. The suspension was then centrifuged (2330 g, 10 min, room temperature). Collect the supernatant in another container. The collected solution was concentrated using an out-of-limit filter unit. The concentrate was added to 20 mM sodium acetate (pH 4.5), centrifuged (2330 g, 10 minutes, room temperature), and concentrated to obtain a 40 kDa protein purified product (Bradford quantification (bovine serum albumin (BSA) conversion), 20.4 mg/mL, 2.21 mL). The purity of the obtained 40 kDa protein was confirmed by SDS-PAGE.

After the 40kDa protein was digested with enzymes, the identification of the protein was attempted by analysis by LC-MS/MS. Cut out the fragment around 40kDa separated by SDS-PAGE, reduce with dithiothreitol (56°C, 1 hour), and carry out carbamoamide methylation with iodoacetamide (protected from light, room temperature) ,45 minutes). Next, 15 μL of 10 ng/μL chymotrypsin solution (5 mM calcium chloride, 50 mM ammonium bicarbonate solution) containing 0.01% ProteaseMax, 15 μL of 5 mM calcium chloride, 50 mM ammonium bicarbonate solution was added, and after overnight incubation, the enzyme digestion was recovered. liquid. The recovered solution was dried under reduced pressure, and then dissolved in 0.1% formic acid solution. This was used in LC-MS/MS analysis.

(Measurement by LC-MS/MS) The measurement by LC-MS/MS was carried out under the following conditions. Equipment used: direct flow nanoLC system Easy-nLC 1000TM (Thermo Scientific) Capture column: Acclaim PepMap (registered trademark) (Thermo Scientific) Analytical column: NANO HPLC CAPILLARY COLUMN (Nikyo-tec Co., Ltd.) Liquid chromatography mass analyzer Q Exactive Plus (Thermo Scientific) Mobile phase: solution A: 0.1% formic acid/water, solution B: 0.1% formic acid/acetonitrile Flow rate: 300nL/min Gradient: 0-40%B/0-30min, 40-60%B/30-35min, 60-90%B/35-37min, 90%B/37-45min Injection volume: 10 μL Ionization Mode: ESI Positive Measuring range: MS1 (m/z 350-1750) Data Dependent Scan Mode

(4) Analysis of protein Protein identification was performed under the following conditions. Search software: Proteome Discoverer 2.2.0.388 (manufactured by ThermoFisher) Biological species: barley (Hordeum vulgare), hops (Humulus), yeast (Saccharomyces cerevisiae) Search terms: Digestive enzyme: chymotrypsin Precursor ion mass error range: monoisotope, ±10ppm Product ion mass error range: ±0.02Da Maximum number of missed cleavages: 5 Percolator: High (the highest degree of accuracy in the three stages of accuracy) Database: SwissProt

As a result, the 40 kDa proteins were found to be Serpin Z4 derived from barley (sequence coverage: 77.2%) and Serpin Z7 derived from barley (sequence coverage: 72.8%).

(Sensory evaluation of adding diketopiperazine to commercially available non-alcoholic beer-flavored beverages) Diketopiperazine was added to a commercially available non-alcoholic beer-flavored beverage, and sensory evaluation of swelling degree was performed. The non-alcoholic beer-flavored beverage is a non-alcohol beer-flavored beverage containing malt as a raw material. The raw materials are malt, hops, carbonic acid, spices, sour, caramel, vitamin C, bitters, and sweeteners. As nutrients, each 100ml contains 0% alcohol, 0g protein, 0g sugar, and dietary fiber. 0~0.1g, about 0mg of purine body.

The standard score of the sensory evaluation is as follows. Five professional evaluators were assigned points on a scale of 0.05 points according to the following criteria, and the score values were averaged. The swelling degree strength is based on the following criteria. 0 points: no feeling at all 1 point: Slightly felt 2 points: clearly felt 3 points: very feeling As the standard score, the commercial beer-flavored alcoholic beverage (I) different from the commercial non-alcoholic beer-flavored beverage described above as the evaluation object was used as the standard beer-flavored alcoholic beverage (I), and the swelling degree was set to 0.7 point. In addition, other commercially available beer-flavored alcoholic beverages were used as the standard beer-flavored alcoholic beverages (II), and the swelling degree was set to 1.5 points of the standard. And the swelling degree of the said commercially available non-alcohol beer-flavored drink which is the evaluation object was made into 0.5 point based on the swelling degree of the said commercially available beer-flavored alcoholic beverages (I) and (II). The standard beer-flavored alcoholic beverage (I) is a beer-flavored alcoholic beverage in which the malt ratio in the raw material exceeds 0% by weight and is less than 50% by weight. The raw materials are sparkling wine, malt, hops, sugar, dietary fiber, and distilled liquor (wheat). As nutrients, each 100ml contains 4% alcohol, 0~0.2g protein, 0.5~0.8g carbohydrate, and purine. About 2.0mg. The standard beer-flavored alcoholic beverage (II) is a beer-flavored alcoholic beverage in which the proportion of malt in the raw material is 50% by weight or more. The raw materials are malt and hops. As nutrients, each 100ml contains 5.5% alcohol, 0.4~0.6g protein, 3.6g carbohydrate, and about 12.5mg purine.

The order of sensory evaluation is as follows. (1) Pour non-alcoholic beer-flavored beverages into 1/10 of the final volume (v/v) small glass bottles (2) The diketopiperazine is weighed by any weight and added (3) Vibration treatment for 30 seconds (4) Stand at room temperature for 30 minutes (5) Fill the non-alcoholic beer-flavored beverage to the final capacity (6) Drinking evaluation after injection

(Analysis of Commercially Available Non-Alcohol Beer-Flavored Beverage) The concentration of diketopiperazine contained in the commercially available non-alcoholic beer-flavored beverage used in the sensory evaluation was quantified by LC-MS in the following order. (1) Preparation of standard product and preparation of calibration curve The diketopiperazine was diluted to the following concentrations, respectively, and passed through a 0.22 μm filter before being supplied for measurement. Final concentration: 0.001ppm, 0.025ppm, 0.050ppm, 0.100ppm, 0.200ppm, 0.300ppm, 0.500ppm, 0.750ppm, 1.000ppm (1ppm=1μg/mL) Diluent use 5% (v/v) ethanol aqueous solution. And, in the analysis result of the standard product, the measured value of the dilution ratio within the range of maintaining the linearity of the calibration curve (R ² >0.99) was used.

The measurement conditions of LC-MS are as follows. LC-MS: X500R manufactured by AB Sciex Separation column: HSST3 1.8μm, 2.1x150mm, manufactured by Waters Esolution: A solution: 0.1% formic acid/water, B solution: 0.1% formic acid/acetonitrile Gradient: A solution: B solution = 98:2→2:98 (27 minutes) Injection volume: 5μL Flow rate: 0.2mL/min Column oven: 40℃ (MS) Ionization Mode: ESI Positive Measurement range: MS1 (m/z 100-1000) Data Independent Scan Mode Ion source temperature: 350℃

(2) Samples for measurement prepared from commercially available non-alcoholic beer-flavored beverages A commercially available non-alcohol beer-flavored beverage was degassed by vibrating, and after the bubbles were relieved, it was appropriately diluted and passed through a 0.22 μm filter before being supplied to the measurement. A 5% (v/v) ethanol aqueous solution was used as the diluent. The concentration of each diketopiperazine contained in a commercially available non-alcoholic beer-flavored beverage was taken as a control group.

(Example 1: Evaluation of Cyclo (Leu-Pro) addition) The concentration of Cyclo (Leu-Pro) contained in a commercially available non-alcoholic beer-flavored beverage was 0.008 ppm. On the other hand, Cyclo (Leu-Pro) was added so that the Cyclo (Leu-Pro) concentration was 0.4 ppm, 1 ppm, 3 ppm, and 10 ppm, respectively, and sensory evaluation was performed. The results of the sensory evaluation are shown in Table 1.

From the results shown in Table 1, it can be seen that in the non-alcoholic beer-flavored beverage containing malt as a raw material, the swelling degree is enhanced when the Cyclo (Leu-Pro) concentration is 0.4 ppm or more.

(Example 2: Evaluation of the additive effect of Cyclo (Leu-Pro) and 40kDa protein) Cyclo (Leu-Pro) was added to commercially available non-alcoholic beer-flavored beverages and the concentration of Cyclo (Leu-Pro) was 0.4ppm as a benchmark, and 40kDa protein was further added to evaluate Cyclo (Leu-Pro) and The additive effect of 40kDa protein. The concentration of 40 kDa protein was 5 ppm. In addition, Cyclo (Leu-Pro) was added to a commercially available non-alcoholic beer-flavored beverage with a concentration of 1 ppm of Cyclo (Leu-Pro) as a reference, and 40 kDa protein was further added to evaluate Cyclo (Leu-Pro) Additive effect with 40kDa protein. The concentration of 40kDa protein was 5ppm, 10ppm, 25ppm. Among the 40kDa proteins, those purified above were used. Furthermore, for comparison, only 40 kDa protein was added to a commercially available non-alcoholic beer flavored beverage, and the evaluation was performed (comparative sample) in which the concentration of 40 kDa protein was 5 ppm. The results of the sensory evaluation are shown in Table 2.

From the results shown in Table 2, it can be seen that the swelling degree can be further enhanced by adding Cyclo (Leu-Pro) to the commercially available non-alcoholic beer-flavored beverage, and further adding 40 kDa protein. From the results of the comparative samples, it can be seen that the swelling degree can be enhanced even by adding only 40 kDa protein. However, the sum of the increase in the control group (0.02) from the sensory evaluation of sample 1 above and the increase in the control group (0.03) from the sensory evaluation of the comparative sample can be found, compared to the combination of diketopiperazine and 40 kDa The additive effect (0.05) in the case of protein was larger than the increase value (0.11) of the control group derived from the sensory evaluation of sample 5. Therefore, the combined use of diketopiperazine and 40 kDa protein exhibited an unexpected additive effect. Moreover, the sum of the increase value (0.12) of the control group from the sensory evaluation of the above-mentioned sample 2 and the increase value (0.03) of the control group from the sensory evaluation of the comparative sample can be obtained, compared with the combination of diketopiperazine and 40kDa The additive effect (0.15) in the case of protein was larger than the increase value (0.20) of the control group derived from the sensory evaluation of sample 6. Therefore, the combined use of diketopiperazine and 40 kDa protein exhibited an unexpected additive effect.

(Example 3: Evaluation of Cyclo (Val-Pro) addition) The concentration of Cyclo (Val-Pro) contained in a commercially available non-alcoholic beer-flavored beverage was 0.012 ppm. On the other hand, Cyclo (Val-Pro) was added so that the Cyclo (Val-Pro) concentration would be 0.4 ppm and 0.8 ppm, respectively, and sensory evaluation was performed. In addition, evaluation was also performed with the concentration of the 40 kDa protein set to 5 ppm. The results of the sensory evaluation are shown in Table 3.

From the results shown in Table 3, it can be seen that the swelling degree can be enhanced by adding Cyclo (Val-Pro) to a commercially available non-alcoholic beer-flavored beverage. Furthermore, by further adding a 40 kDa protein, the degree of swelling can be further enhanced. From the results of the comparative samples (refer to Table 2), it can be seen that the swelling degree can be enhanced even by adding only 40 kDa protein. However, the sum of the increase value (0.02) of the control group from the sensory evaluation of the above-mentioned sample 9 and the increase value (0.03) of the control group from the sensory evaluation of the comparative sample shown in Table 2 can be obtained, compared with the combined use of the two The additive effect of ketopiperazine and 40kDa protein (0.05), the increase value (0.09) of the control group from the functional evaluation of sample 10 is larger, so by using diketopiperazine and 40kDa protein together, it can exert unexpected effects. Multiplier effect. Moreover, the sum of the increase value (0.08) of the control group from the sensory evaluation of the above-mentioned sample 11 and the increase value (0.03) of the control group from the sensory evaluation of the comparative sample shown in Table 2 can be obtained. The additive effect of ketopiperazine and the 40kDa protein (0.11) was greater in the control group (0.20) from the functional evaluation of sample 12. Therefore, by using the diketopiperazine and the 40kDa protein in combination, an unexpected effect could be exerted. Multiplier effect.

(Example 4: Evaluation of Cyclo (Ile-Pro) addition) The concentration of Cyclo (Ile-Pro) contained in commercially available non-alcoholic beer-flavored beverages is 0 ppm. On the other hand, Cyclo (Ile-Pro) was added so that the Cyclo (Ile-Pro) concentration would be 0.4 ppm and 0.8 ppm, respectively, and sensory evaluation was performed. In addition, evaluation was also performed with the concentration of the 40 kDa protein set to 5 ppm. The results of the sensory evaluation are shown in Table 4.

From the results shown in Table 4, it can be seen that the swelling degree can be enhanced by adding Cyclo (Ile-Pro) to a commercially available non-alcoholic beer-flavored beverage. Furthermore, by further adding a 40 kDa protein, the degree of swelling can be further enhanced. From the results of the comparative samples (refer to Table 2), it can be seen that the swelling degree can be enhanced even by adding only 40 kDa protein. However, the sum of the increase value (0.04) of the control group from the sensory evaluation of the above-mentioned sample 13 and the increase value (0.03) of the control group from the sensory evaluation of the comparative sample shown in Table 2 can be obtained, compared with the combined use of the two The additive effect of ketopiperazine and 40kDa protein (0.07), the increase value (0.11) of the control group from the functional evaluation of sample 14 is larger, so by using diketopiperazine and 40kDa protein together, it is possible to exert unexpected effects. Multiplier effect. Moreover, the sum of the increase value (0.09) of the control group from the sensory evaluation of the above-mentioned sample 15 and the increase value (0.03) of the control group from the sensory evaluation of the comparative sample shown in Table 2 can be obtained. The additive effect of ketopiperazine and 40kDa protein (0.12), the increase value (0.21) of the control group from the functional evaluation of sample 16 is larger, so by using diketopiperazine and 40kDa protein together, it is possible to exert unexpected effects. Multiplier effect. [industrial availability]

According to the present invention, it is possible to provide a beverage with enhanced swelling among non-alcoholic beer-flavored beverages containing malt as a raw material.

Claims (4)

A non-alcoholic beer-flavored beverage, which is a non-alcoholic beer-flavored beverage containing malt in raw materials, and comprises a non-alcoholic beer-flavored beverage selected from the group consisting of: At least one kind of diketopiperazine in the group of amino acids, and among the above-mentioned diketopiperazine, the concentration of the most contained diketopiperazine is 0.4 ppm or more. The non-alcohol beer-flavored beverage according to claim 1, wherein among the diketopiperazines, the concentration of the diketopiperazine most contained is 10 ppm or less. The non-alcoholic beer-flavored beverage according to claim 1 or 2, further comprising a protein with a molecular weight of 35 to 50 kDa, and the concentration of the protein is 5 ppm or more. The non-alcoholic beer-flavored beverage according to claim 3, wherein the protein concentration is 30 ppm or less.