JP2016123400A - Beer-taste alcoholic beverage and production method therefor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a beer-taste alcoholic beverage that has a suppressed miscellaneous taste, a beer-like soft and smooth texture, and a balanced taste of harmony.SOLUTION: Provided is a beer-taste fermented alcoholic beverage produced from malt and/or non-germination barley as a part of the raw-material, and in which the concentration of a peptide having a molecular weight of 10 to 20 kDa (gel permeation method) is 0.15 mg/mL or more.SELECTED DRAWING: None

Description

  The present invention relates to a beer-taste fermented alcoholic beverage made of malt and / or ungerminated barley as a raw material and a method for producing the same. The present invention also provides a flavor improving agent for a beer-taste alcoholic beverage comprising a peptide having a specific molecular weight derived from a beer-taste fermented alcoholic beverage made from malt and / or ungerminated wheat as a raw material, and the peptide. It also relates to a method for improving the flavor of the used beer-taste alcoholic beverage.

  Beer-taste alcoholic beverages such as sparkling liquor and new genre beverages are cheaper than beer and are widely drunk as an alternative to beer. However, these beverages have insufficient soft and smooth textures such as beer, have a lot of miscellaneous tastes such as astringency and roughness, and lack in harmony with the flavor, and there are points to be improved in flavor.

  As a technique for imparting richness and taste thickness to a beer-taste alcoholic beverage, techniques (Patent Documents 1 and 2) that contain carboxymethyllysine and use a peptide as a rich imparting substance are known. However, there is still room for improvement in the flavor of beer-taste alcoholic beverages, in particular, the application of soft and smooth textures, suppression of miscellaneous taste, and a sense of harmony in taste.

JP 2011-227070 A JP 2014-158502 A

  It is an object of the present invention to provide a beer-taste alcoholic beverage having a soft and smooth texture like beer, having a harmonious taste, and a method for producing the same. Another object of the present invention is to provide a flavor improving agent and a flavor improving method for beer-taste alcoholic beverages.

  The present inventors have found that in a beer-taste alcoholic beverage, a peptide having a specific molecular weight and α-glucan having a specific polymerization degree contribute to imparting a soft and smooth texture like beer and reducing miscellaneous taste. The present inventors have also found that a more harmonious taste like beer can be realized by setting the concentration of a peptide having a specific molecular weight within a specific concentration range. Furthermore, the present inventors have found that the above-described effect can be further exerted by setting the concentration of α-glucan having a specific polymerization degree in a specific concentration range in addition to a peptide having a specific molecular weight. The inventors further specifically identified peptides that contribute to the flavor improvement of beer-taste alcoholic beverages. The present invention is based on these findings.

According to the present invention, the following inventions are provided.
(1) A beer-taste fermented alcoholic beverage having malt and / or ungerminated wheat as part of the raw material, and having a molecular weight of 10 to 20 kDa (gel filtration method) and a peptide concentration of 0.15 mg / ml or more, Beer-taste fermented alcoholic beverage.
(2) The beer-taste fermented alcoholic beverage according to (1) above, wherein the malt use ratio is less than two-thirds.
(3) The beer-taste fermented alcoholic beverage according to (1) or (2) above, wherein the ratio of the peptide amount having a molecular weight of 10 to 20 kDa to the total protein amount is larger than 3.6%.
(4) The beer-taste fermented alcoholic beverage according to any one of (1) to (3) above, wherein the peptide concentration of a molecular weight of 10 to 20 kDa is 0.19 mg / ml or more.
(5) The beer-taste fermented alcoholic beverage according to any one of (1) to (4) above, wherein the concentration of α-glucan having a degree of polymerization of 5 to 10 is 2.1 mg / ml or more.
(6) The beer-taste fermented alcoholic beverage according to (5) above, wherein the concentration of α-glucan having a polymerization degree of 5 to 10 is 3.3 mg / ml or more.
(7) One or more peptides having a molecular weight of 10 to 20 kDa (gel filtration method) derived from a beer-taste fermented alcoholic beverage that uses malt and / or ungerminated wheat as a raw material are blended. , Beer-taste alcoholic beverages.
(8) The beer-taste alcoholic beverage according to (7), wherein the peptide having a molecular weight of 10 to 20 kDa is blended so as to have a concentration of 0.15 mg / ml or more.
(9) The beer-taste alcoholic beverage according to (7) or (8), wherein the peptide having a molecular weight of 10 to 20 kDa is blended so as to have a ratio of more than 3.6% with respect to the total protein amount in the beverage.
(10) One or two or more kinds of α-glucan having a polymerization degree of 5 to 10 derived from beer-taste fermented alcoholic beverages that use malt and / or ungerminated wheat as a raw material are further blended, The beer-taste alcoholic drink in any one of said (7)-(9).
(11) The beer-taste alcoholic beverage according to (10), wherein α-glucan having a polymerization degree of 5 to 10 is blended so as to have a concentration of 2.1 mg / ml or more.
(12) One or more peptides having a molecular weight of 10 to 20 kDa are selected from the group consisting of α-amylase / trypsin inhibitor, serpin-Z4 and non-specific lipid-transfer protein 1 The beer-taste alcoholic drink in any one of said (7)-(11) which is 1 type, or 2 or more types selected.
(13) A step of blending one or more peptides having a molecular weight of 10 to 20 kDa (gel filtration method) derived from a beer-taste fermented alcoholic beverage that uses malt and / or ungerminated wheat as part of the raw material. A method for producing a beer-taste alcoholic beverage.
(14) The method for producing a beer-taste alcoholic beverage according to (13), wherein the peptide having a molecular weight of 10 to 20 kDa is blended so as to have a concentration of 0.15 mg / ml or more.
(15) The beer-taste alcoholic beverage according to (13) or (14), wherein the peptide having a molecular weight of 10 to 20 kDa is blended so as to have a ratio of more than 3.6% with respect to the total amount of protein in the beverage. Production method.
(16) including a step of further blending one or two or more α-glucans having a degree of polymerization of 5 to 10 derived from a beer-taste fermented alcoholic beverage that uses malt and / or ungerminated wheat as part of the raw material The manufacturing method of the beer taste alcoholic beverage in any one of said (13)-(15) which consists of.
(17) The method for producing a beer-taste alcoholic beverage according to (16), wherein α-glucan having a polymerization degree of 5 to 10 is blended so as to have a concentration of 2.1 mg / ml or more.
(18) One or more peptides having a molecular weight of 10 to 20 kDa are selected from the group consisting of α-amylase / trypsin inhibitor, serpin-Z4 and non-specific lipid-transfer protein 1 The manufacturing method of the beer taste alcoholic beverage in any one of said (13)-(17) which is 1 type, or 2 or more types selected.
(19) As an active ingredient, one or more peptides having a molecular weight of 10 to 20 kDa (gel filtration method) derived from a beer-taste fermented alcoholic beverage that uses malt and / or ungerminated wheat as a raw material A flavor improving agent for beer-taste alcoholic beverages.
(20) A step of adding one or more peptides having a molecular weight of 10 to 20 kDa (gel filtration method) derived from a beer-taste fermented alcoholic beverage that uses malt and / or ungerminated wheat as part of the raw material. A method for improving the flavor of a beer-taste alcoholic beverage.

  According to the present invention, a peptide having a molecular weight of 10 to 20 kDa derived from a beer-taste fermented alcoholic beverage containing malt and / or ungerminated wheat as a raw material is blended, or the concentration of the peptide is within a predetermined range. By making it inside, it is possible to provide a beer-taste alcoholic beverage that has a soft and smooth texture like beer and has a harmonious taste. In particular, beer-taste fermented alcoholic beverages with a malt use ratio of less than 50% may not have the taste (especially soft and smooth texture) that can be felt with beer, and there is an improvement in flavor (astringency and roughness) The present invention is advantageous in that the flavor of such a beverage as a beer-taste beverage can be improved or improved. . Moreover, since a peptide having a molecular weight of 10 to 20 kDa is present in a beer-taste fermented alcoholic beverage that uses malt or ungerminated wheat as a raw material, the peptide is added to the beer-taste alcoholic beverage and blended. However, it is also advantageous in that no off-flavors are produced.

It is the figure which showed the amount of peptides added to beer type malt alcoholic beverage in Example 1 (6). It is the figure which showed the alpha-glucan amount added to beer type malt alcoholic beverage in Example 1 (6). It is the figure which showed the sensory evaluation result of Example 1 (6). A sensory evaluation score was shown for each added fraction. The sensory evaluation score of the additive-free control was 2.5. It is the figure which showed the analytical curve created from the retention time of the HPLC gel filtration fraction implemented in Example 2 (3), and the molecular weight of a known substance. It is a bubble graph which showed the sensory evaluation result (taste like beer) of Example 2 (4). Each sample was plotted with the degree of polymerization being 5 to 10 α-glucan concentration (mg / ml) and the horizontal axis being the 10 to 20 kDa peptide concentration (mg / ml). The horizontal stripe pattern corresponds to a commercial product, the vertical stripe pattern corresponds to less than 50% malt (test brew), and the rhombus pattern corresponds to less than 25% malt (test brew). The bubble size represents a beer-like taste score. It is a bubble graph which showed the sensory evaluation result (taste like beer) of Example 2 (4). Each sample was plotted with the vertical axis representing the 10-20 kDa peptide ratio (%) and the horizontal axis representing the 10-20 kDa peptide concentration (mg / ml). The horizontal stripe pattern corresponds to a commercial product, the vertical stripe pattern corresponds to less than 50% malt (test brew), and the rhombus pattern corresponds to less than 25% malt (test brew). The bubble size represents a beer-like taste score. It is the figure which evaluated the correlation with the alpha-glucan density | concentration of 5-10 degrees of polymerization, the peptide density | concentration of 10-20 kDa, and a sensory evaluation result. FIG. 7A is a graph plotting sensory evaluation results against α-glucan concentrations having a degree of polymerization of 5 to 10. FIG. 7B is a diagram plotting sensory evaluation results against a peptide concentration of 10 to 20 kDa. FIG. 7C is a graph plotting sensory evaluation results against a value obtained by multiplying an α-glucan concentration having a polymerization degree of 5 to 10 by a peptide concentration of 10 to 20 kDa. It is the bubble graph which showed the sensory evaluation result (taste like beer) of Example 4 (2). Each sample was plotted with the degree of polymerization being 5 to 10 α-glucan concentration (mg / ml) and the horizontal axis being the 10 to 20 kDa peptide concentration (mg / ml). The horizontal stripe pattern corresponds to the sample added to sample No. 4, the vertical stripe pattern corresponds to the sample added to sample No. 19, and the rhombus pattern corresponds to the sample added to sample No. 16. The bubble size represents a beer-like taste score. It is the bubble graph which showed the sensory evaluation result (taste like beer) of Example 4 (2). Each sample was plotted with the vertical axis representing the 10-20 kDa peptide ratio (%) and the horizontal axis representing the 10-20 kDa peptide concentration (mg / ml). The horizontal stripe pattern corresponds to the sample added to sample No. 4, the vertical stripe pattern corresponds to the sample added to sample No. 19, and the rhombus pattern corresponds to the sample added to sample No. 16. The bubble size represents a beer-like taste score. It is the graph which showed the sensory evaluation result of Example 4 (2). FIG. 10A is a sensory evaluation result about the taste like beer. FIG. 10B shows the sensory evaluation results for the roughness remaining in the mouth. It is the figure which showed the result of 2D-PAGE electrophoresis about the drink manufactured in Example 1 and 2. FIG. STD is a molecular weight marker. FIG. 11A shows the results for the test section 1 of Example 1, and FIG. 11B shows the results for the test section 2 of Example 1. FIG. 11C shows the results for the sample 16 of Example 2, and FIG. 11D shows the results for the sample 15 of Example 2.

Detailed description of the invention

  In the present specification, “beer-taste alcoholic beverage” means an alcoholic beverage having a taste and aroma peculiar to beer obtained when beer is usually produced, that is, when beer is produced based on fermentation by yeast or the like. .

  Beer-taste alcoholic beverages also include “beer-taste fermented alcoholic beverages” fermented by yeast using carbon sources, nitrogen sources, and water as raw materials, and “beer-taste fermented alcoholic beverages” include beer, foam Beer-taste sparkling alcoholic beverages that do not use liquor, wheat or malt as raw materials (for example, brewing new genre beverages classified as “Other brewed liquor (foaming) (1)” under the liquor tax law) and malt as raw materials Examples include beverages obtained by adding alcohol to beer or sparkling liquor (for example, liqueur-based new genre beverages classified as “liqueur (sparkling) (1)” in the liquor tax law).

  According to the first aspect of the present invention, there is provided a beer-taste fermented alcoholic beverage using malt and / or ungerminated wheat as a part of the raw material. The beer-taste fermented alcoholic beverage of the present invention can use at least malt as a raw material derived from wheat. In that case, the malt use ratio can be less than 2/3, and preferably the malt use ratio is Less than 50%. In the present specification, the “malt use ratio” refers to the ratio of malt mass to the mass of all raw materials excluding brewing water.

  The beer-taste fermented alcoholic beverage of the present invention is characterized in that the peptide concentration having a molecular weight of 10 to 20 kDa derived from the raw material of the beer-taste fermented alcoholic beverage is a specific value or more. In this specification, the “peptide concentration” is calculated by summing the contents of one or more peptides having a molecular weight of 10 to 20 kDa. In the present specification, the “molecular weight” of a peptide is measured by gel filtration, and a specific example of the measurement is as shown in Example 2 below. Peptide quantification can be performed by the Lowry method.

  In the beer-taste fermented alcoholic beverage of the present invention, the peptide concentration with a molecular weight of 10 to 20 kDa can be 0.15 mg / ml or more, preferably 0.19 mg / ml or more, more preferably 0.20 mg / ml or more. The peptide concentration can have an upper limit from the viewpoint of harmony of taste, and for example, the upper limit can be 0.45 mg / ml. Further, the ratio of the amount of peptide having a molecular weight of 10 to 20 kDa to the total amount of protein can be made larger than 3.6%, preferably 3.7% or more, and more preferably 4.0% or more. The upper limit of the ratio can be set from the viewpoint of harmony of taste, and for example, the upper limit can be 8.0%. By setting the concentration and ratio of the peptide having a molecular weight of 10 to 20 kDa to a specific value or more as shown in Examples 2 and 4 to be described later, a beer taste fermented alcoholic beverage (particularly, a beer taste having a malt use ratio of less than 50%) Fermented alcoholic beverage) can be reduced, and the beverage can be a beverage having a soft and smooth texture like beer and having a harmonious taste.

  The beer-taste fermented alcoholic beverage of the present invention is also characterized in that the concentration of α-glucan having a polymerization degree of 5 to 10 is a specific value or more. In the present specification, “α-glucan concentration” is calculated by summing the contents of one or more α-glucans having a degree of polymerization of 5 to 10. In the present specification, “α-glucan” means a linear or branched glucan composed of a plurality of glucose molecules bonded by α-1,4-glucoside bonds. Furthermore, in this specification, “degree of polymerization” of α-glucan means the number of glucose residues constituting glucan, and not only the number of glucose residues constituting linear glucan but also glucose constituting a branched structure. Contains the number of residues. The degree of polymerization and content of α-glucan can be measured by LC-MS / MS (liquid chromatography-mass spectrometry), and a specific example of the measurement is as shown in Example 2 below. In the present specification and drawings, the degree of polymerization may be simply expressed as “G”.

  In the beer-taste fermented alcoholic beverage of the present invention, the concentration of α-glucan having a polymerization degree of 5 to 10 can be 2.1 mg / ml or more, preferably 3.3 mg / ml or more, more preferably 3.9 mg / ml or more. It is. The α-glucan concentration can have an upper limit from the viewpoint of harmony of taste, for example, 13.0 mg / ml can be set as the upper limit. As shown in Examples 2 and 4 below, by setting the α-glucan concentration having a polymerization degree of 5 to 10 to a specific value or more, a beer-taste fermented alcoholic beverage (particularly, a beer taste having a malt use ratio of less than 50%) Fermented alcoholic beverage) can be reduced, and the beverage can be a beverage having a soft and smooth texture like beer and having a harmonious taste.

  In the beer-taste fermented alcoholic beverage according to the present invention, the α-glucan concentration having a degree of polymerization of 5 to 10 is added in addition to the peptide concentration of molecular weight of 10 to 20 kDa from the viewpoint of further suppressing miscellaneous taste and further harmony of texture and taste Is preferably a predetermined value. In the beer-taste fermented alcoholic beverage of the present invention, the miscibility is further suppressed by adjusting the peptide having a molecular weight of 10 to 20 kDa to a predetermined concentration and adjusting the α-glucan concentration having a polymerization degree of 5 to 10 to a predetermined concentration. It has not been reported so far that the harmony of texture and taste is further improved. That is, in the beer-taste fermented alcoholic beverage of the present invention, the peptide concentration with a molecular weight of 10 to 20 kDa is 0.15 mg / ml or more (preferably 0.19 mg / ml or more, more preferably 0.20 mg / ml or more), and The concentration of α-glucan having a degree of polymerization of 5 to 10 can be 2.1 mg / ml or more (preferably 3.3 mg / ml or more, more preferably 3.9 mg / ml or more), and the molecular weight relative to the total protein amount The ratio of the peptide amount of 10 to 20 kDa can be made larger than 3.6% (preferably 3.7% or more, more preferably 4.0% or more). The peptide concentration, α-glucan concentration, and peptide ratio can all have an upper limit as described above.

  As long as the beer-taste fermented alcoholic beverage of the present invention has a peptide concentration of molecular weight of 10 to 20 kDa and / or an α-glucan concentration of a polymerization degree of 5 to 10 within a predetermined range, production of a normal beer-taste fermented alcoholic beverage It can be manufactured according to the procedure. For example, fermented brewer's yeast can be added to wort prepared from brewing raw materials such as malt, hops, auxiliary raw materials, and brewing water for fermentation, and a fermented malt beverage can be brewed. The obtained beer-taste fermented alcoholic beverage can be stored at a low temperature, and then the yeast can be removed by a filtration process.

  In the above production procedure, wort can be produced according to a conventional method. For example, saccharifying a mixture of brewing raw materials and brewing water, filtering to obtain wort, adding hops to the wort, boiling, and preparing the wort by cooling the boiled wort Can do. Moreover, wort can also be produced by adding a commercially available enzyme preparation during the saccharification step. For example, protease preparations for proteolysis, α-amylase preparations, glucoamylase preparations, pullulanase preparations, etc. for carbohydrate degradation, β-glucanase preparations, fibrinolytic enzyme preparations, etc. for fibrinolysis, respectively. These can be used, or these mixed preparations can also be used.

  In the production of the beer-taste fermented alcoholic beverage of the present invention, in addition to malt, ungerminated wheat (eg, ungerminated barley (including extracted), ungerminated wheat (including extracted))); rice , Corn, corn, potato, starch, starch, sugar (eg, liquid sugar) and other auxiliary materials specified by the liquor tax law; nitrogen sources such as proteolysates and yeast extract; fragrances, pigments, foaming / foaming improver, water quality Other additives such as regulators and fermentation aids can be used as brewing raw materials. That is, the beer-taste fermented alcoholic beverage of the present invention can use raw materials other than brewing water at least as malt, ungerminated barley (preferably ungerminated barley), and hops. Corn, starch, etc. can be used as raw materials.

  In order to adjust the peptide concentration of a molecular weight of 10 to 20 kDa in the produced beverage in the method for producing a beer-taste fermented alcoholic beverage of the present invention within a predetermined value range, for example, malt and / or ungerminated wheat It can be adjusted by suppressing proteolysis in the preparation and saccharification process or by suppressing the proteolysis degree in the malting process of malt as a raw material. Examples of proteolysis include proteases inherent in malt and ungerminated germs, or protease preparations added from the outside. Degradation can be suppressed by reducing the amount of protease preparation added, reducing the treatment time at the proteolytic action temperature, or changing the working pH from the optimum condition.

  In order to adjust the concentration of α-glucan having a degree of polymerization of 5 to 10 in the produced beverage in the method for producing a beer-taste fermented alcoholic beverage of the present invention within a predetermined value range, for example, malt and / or ungerminated Inhibiting α-glucan degradation in the preparation and saccharification process of wheat, inhibiting α-glucan degradation in the malting process of malt as a raw material, or using liquid sugar in which the degree of degradation of α-glucan is suppressed It can be adjusted depending on the situation. Examples of α-glucan degradation include α-amylase and β-amylase inherent in malt and ungerminated wheat, or α-amylase preparations, glucoamylase preparations, pullulanase preparations, etc. added from the outside. Degradation can be suppressed by reducing the amount of α-amylase preparation, glucoamylase preparation, pullulanase preparation, etc., reducing the treatment time at the action temperature of α-glucan decomposition, changing the working pH from the optimum condition, etc. .

  Alternatively, as described in Example 4, a beer-taste fermented alcoholic beverage is prepared according to the procedure described in Example 2, and a peptide having a molecular weight of 10-20 kDa is included from the beverage according to the procedure described in Example 1. And a fraction containing an α-glucan having a polymerization degree of 5 to 10 and a fraction containing a peptide having a molecular weight of 10 to 20 kDa and / or a polymerization degree of 5 by adding the fraction to a beer-taste fermented alcoholic beverage. It is also possible to produce a beer-taste fermented alcoholic beverage having an α-glucan concentration of 10 to 10 adjusted within a predetermined value range.

  According to the second aspect of the present invention, one or more peptides having a molecular weight of 10 to 20 kDa derived from beer-taste fermented alcoholic beverages that use malt and / or ungerminated wheat as a raw material are blended. A beer-taste alcoholic beverage and a method for producing the beverage are provided. The beverage containing the peptide has improved or improved flavor as a beer-taste alcoholic beverage. Specifically, the miscellaneous taste is suppressed and there is a soft and smooth texture like beer, which is harmonious. It is a delicious beverage.

  Examples of one or two or more peptides having a molecular weight of 10 to 20 kDa derived from a beer-taste fermented alcoholic beverage made from malt and / or ungerminated wheat as raw materials include α-amylase / trypsin inhibitor, serpin -Z4 and non-specific lipid-transfer protein 1 are preferred, preferably these proteins and peptides are derived from barley. When α-amylase / trypsin inhibitor, serpin-Z4 or non-specific lipid transfer protein 1 is added to a beer-taste alcoholic beverage, these peptides or proteins are malt or ungerminated wheat beer as part of the raw material. Peptides or proteins other than those prepared from taste fermented alcoholic beverages may be used.

  The peptide concentration of the molecular weight of 10 to 20 kDa in the beer-taste alcoholic beverage of the present invention can be 0.15 mg / ml or more, preferably 0.19 mg / ml or more, more preferably 0.20 mg / ml or more. The peptide concentration can have an upper limit from the viewpoint of harmony of taste, and for example, the upper limit can be 0.45 mg / ml. Further, the ratio of the peptide amount having a molecular weight of 10 to 20 kDa with respect to the total protein amount can be made larger than 3.6%, preferably 3.7% or more, more preferably 4.0% or more. The upper limit of the ratio can be set from the viewpoint of harmony of taste, and for example, the upper limit can be 8.0%. By setting the concentration and ratio of the peptide having a molecular weight of 10 to 20 kDa to a specific value or more as shown in Examples 2 and 4 to be described later, a beer taste fermented alcoholic beverage (particularly, a beer taste having a malt use ratio of less than 50%) Fermented alcoholic beverage) can be reduced, and the beverage can be a beverage having a soft and smooth texture like beer and having a harmonious taste. For the measurement of the molecular weight and content of a peptide having a molecular weight of 10 to 20 kDa, reference can be made to the description of the beer-taste fermented alcoholic beverage that is the first aspect of the present invention.

  In the beer-taste alcoholic beverage of the present invention, one or two or more α-glucans having a degree of polymerization of 5 to 10 derived from a beer-taste fermented alcoholic beverage made of malt and / or ungerminated wheat are used as raw materials. Furthermore, you may mix | blend. Examples of one or two or more kinds of α-glucan having a degree of polymerization of 5 to 10 derived from beer-taste fermented alcoholic beverages that use malt and / or ungerminated wheat as a raw material include maltopentaose, malto Hexaose, maltoheptaose, maltooctaose, maltononaose and maltodechaose. When α-glucan is blended in a beer-taste alcoholic beverage, α-glucan is an α-glucan other than those prepared from beer-taste fermented alcoholic beverages that are made from malt and / or ungerminated wheat. Further, it may be a fraction obtained by fractionating a starch degradation product obtained by allowing an enzyme to act on starch.

  The α-glucan concentration with a degree of polymerization of 5 to 10 can be 2.1 mg / ml or more, preferably 3.3 mg / ml or more, more preferably 3.9 mg / ml or more. The α-glucan concentration can have an upper limit from the viewpoint of harmony of taste, for example, 13.0 mg / ml can be set as the upper limit. As shown in Examples 2 and 4 below, by setting the α-glucan concentration having a polymerization degree of 5 to 10 to a specific value or more, a beer-taste fermented alcoholic beverage (particularly, a beer taste having a malt use ratio of less than 50%) Fermented alcoholic beverage) can be reduced, and the beverage can be a beverage having a soft and smooth texture like beer and having a harmonious taste. For the measurement of the polymerization degree and content of α-glucan having a polymerization degree of 5 to 10, the description of the beer-taste fermented alcoholic beverage which is the first aspect of the present invention can be referred to.

  In the beer-taste alcoholic beverage of the present invention, from the viewpoint of further suppressing miscellaneous taste and further harmony of texture and taste, in addition to a peptide having a molecular weight of 10 to 20 kDa, an α-glucan having a polymerization degree of 5 to 10 is blended. It is preferable. In the beer-taste fermented alcoholic beverage of the present invention, in addition to a peptide having a molecular weight of 10 to 20 kDa, an α-glucan having a polymerization degree of 5 to 10 is further suppressed, and the texture and taste are further harmonized. It has not been reported so far. That is, in the beer-taste alcoholic beverage of the present invention, the peptide concentration with a molecular weight of 10 to 20 kDa is 0.15 mg / ml or more (preferably 0.19 mg / ml or more, more preferably 0.20 mg / ml or more), and polymerization is performed. The α-glucan concentration at a degree of 5 to 10 can be 2.1 mg / ml or more (preferably 3.3 mg / ml or more, more preferably 3.9 mg / ml or more), and the molecular weight relative to the total protein amount is 10 The ratio of the amount of peptide of ˜20 kDa can be made larger than 3.6% (preferably 3.7% or more, more preferably 4.0% or more). The peptide concentration, α-glucan concentration, and peptide ratio can all have an upper limit as described above.

  One or more peptides having a molecular weight of 10 to 20 kDa and α-glucan having a polymerization degree of 5 to 10 to be blended in a beer-taste alcoholic beverage are malt and / or ungerminated wheat as part of the raw material It can be prepared from a taste fermented alcoholic beverage (preferably a beer-taste fermented alcoholic beverage that uses at least malt as a raw material derived from wheat). For example, a beer-taste fermented alcoholic beverage is produced according to the procedure described in Example 1 or Example 2, and a molecular weight of 10 is obtained from the beverage using HPLC gel filtration fractionation or a solid phase extraction column as described in Example 1. A fraction containing a peptide of ˜20 kDa and a fraction containing α-glucan having a polymerization degree of 5 to 10 can be prepared. The peptide having a molecular weight of 10 to 20 kDa and the α-glucan having a polymerization degree of 5 to 10 are not necessarily isolated and purified, and the peptide fraction and α-glucan obtained by fractionation treatment from a beer-taste fermented alcoholic beverage Fractions can be blended into beer-taste alcoholic beverages.

  Blending into a beer-taste alcoholic beverage can be performed by addition to a pre-fermentation liquid before fermentation, a fermentation liquid during fermentation, or a fermentation liquid after fermentation. For example, as described in Example 4, a beer-taste fermented alcoholic beverage is produced according to the procedure described in Example 2, and a peptide having a molecular weight of 10-20 kDa is included from the beverage according to the procedure described in Example 1. And a peptide containing α-glucan having a polymerization degree of 5 to 10, and a peptide having a molecular weight of 10 to 20 kDa was blended by adding the fraction to a beer-taste fermented alcoholic beverage after fermentation. A beer-taste alcoholic beverage can be produced.

  The beer-taste alcoholic beverage of the present invention should be produced according to the usual procedure for producing a beer-taste alcoholic beverage, except that a peptide concentration of molecular weight of 10 to 20 kDa and an α-glucan concentration of a polymerization degree of 5 to 10 are optionally blended. Can do. For example, fermented brewer's yeast can be added to wort prepared from brewing raw materials such as malt, hops, auxiliary raw materials, and brewing water for fermentation, and a fermented malt beverage can be brewed. The obtained beer-taste fermented alcoholic beverage can be stored at a low temperature, and then the yeast can be removed by a filtration step.

  In the above production procedure, wort can be produced according to a conventional method. For example, saccharifying a mixture of brewing raw materials and brewing water, filtering to obtain wort, adding hops to the wort, boiling, and preparing the wort by cooling the boiled wort Can do.

  In the production of the beer-taste alcoholic beverage of the present invention, raw materials other than malt can be used. Specifically, raw materials other than malt can be used according to the description regarding the beer-taste fermented alcoholic beverage of the first aspect of the present invention.

  As mentioned above, although demonstrated centering on the beer taste alcoholic beverage which is the 2nd surface of this invention, the manufacturing method of the beer taste alcoholic beverage of this invention can be implemented according to the said description about this drink.

  According to the third aspect of the present invention, one or more peptides having a molecular weight of 10 to 20 kDa derived from a beer-taste fermented alcoholic beverage that uses malt and / or ungerminated wheat as a raw material are effective. A flavor improving agent for a beer-taste alcoholic beverage and a method for improving the flavor of the beverage are provided. In the present specification, “improvement of flavor of a beer-taste alcoholic beverage” means that a miscellaneous taste is suppressed and a soft and smooth texture like beer and a harmonious taste are more felt. The flavor improving agent and flavor improving method of the present invention can be carried out according to the description of the beer-taste alcoholic beverage and the method for producing the beverage which are the second aspect of the present invention.

  The present invention will be described more specifically based on the following examples, but the present invention is not limited to these examples. In the following examples, the percentage (%) represents mass% unless otherwise specified.

Example 1: Addition of peptide fraction and α-glucan fraction to beer-taste alcoholic beverage and sensory evaluation (Part 1)
(1) Production of beer-taste fermented alcoholic beverage A beer-taste fermented alcoholic beverage was produced by the infusion method using barley malt, hops, and enzyme preparation.

  In test group 1, 100 g of barley malt was added to 300 ml of hot water at 50 ° C., the enzyme preparation was added and held for 60 minutes, then heated to 65 ° C. and held for 60 minutes, and further heated to 78 ° C. for 5 minutes. After holding, it was filtered to obtain wort. In test group 2, wort was similarly obtained except that the 50 ° C. process was not performed and the enzyme preparation was not added. Subsequently, 0.8 g / L of hops was added and boiled at 100 ° C. for 90 minutes, followed by filtration to obtain a pre-fermentation solution.

  Then, it fermented with the brewer's yeast according to the conventional method, and the flavor check of the fermented liquid was performed. The panel of 8 people, "softness of taste", that is, the miscellaneous taste is suppressed, there is a soft and smooth texture like beer, and there is a harmonious taste, the minimum is 1 point, the maximum is 5 points As a result, sensory evaluation was performed in five stages, and an average score was calculated. The results were as shown in Table 1.

  As shown in Table 1, the test group 2 had a better sensory evaluation score (softness of taste) than the test group 1, and the flavor impression was also preferred.

(2) Gel filtration fraction The fermentation broth obtained in (1) 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 fractionation sample. The sample was subjected to gel filtration fractionation under the following conditions.

Column: Hiload Superdex 30pg 26/600 (manufactured by GE Healthcare)
Sample injection volume: 5ml
Eluent composition: 100 mM NaCl
Flow rate: 2.5 mL / min (constant flow rate)
Detection wavelength: 215 nm
Preparative: 0.29 cv (column volume) to 19.1 ml (fraction 0),
Then fractions from 0.35 cv to 5 ml each (fraction 1 to 51)

  According to the sensory evaluation of the fractions, the fractions were divided into nine groups of pre-fractions, A1, A2, B, C, D, E, F, and G, as shown in Table 2, depending on the flavor characteristics.

(3) Purification of peptide fraction and α-glucan fraction Each fraction obtained in (2) above was obtained from a solid phase extraction column (fractions A1, A2, and B were Bond Elute C18 EWP, fraction C, D, E, F, and G use Bond Elute C18, manufactured by Agilent Technologies), washed with demineralized water, eluted with 50% ethanol aqueous solution, concentrated and dried, Was condensed with demineralized water to obtain a concentrated solution. This was used as a peptide fraction. The flow-through fraction of the solid phase extraction column was further desalted with anion, cation, ion exchange resin (Amberlite IR120H and Amberlite XE583, manufactured by Organo), deodorized with a small amount of activated carbon, and concentrated to dryness. Solidified and condensed with demineralized water to obtain a concentrated solution. This was designated as α-glucan fraction.

(4) Protein quantification by the Lowry method The protein quantification of the peptide fraction obtained by the gel filtration fraction of (3) above was performed by the Lowry method using a commercially available kit (DC protein assay, manufactured by Bio-Rad). . First, 50 μL of the above fraction solution was taken and centrifuged to dryness, and 10 μL of ultrapure water was added and redissolved to obtain an analysis sample. Thereto, 50 μL of solution A was added and stirred, and then 400 μL of solution B was added and stirred. After a color development reaction at room temperature for 15 minutes, 350 μL was transferred to a 96-well plate and the absorbance at 750 nm was measured. The amount of peptide was calculated based on the obtained absorbance and a calibration curve prepared in advance. The calibration curve was prepared using BSA (bovine serum albumin).

(5) Quantification of α-glucan Quantification of α-glucan contained in the α-glucan fraction obtained in the above (3) was carried out using glucoamylase and a buffer (final concentration 10 U / ml, 100 mM Na acetate pH 4.5). ) And reacted at 40 ° C. overnight, and then measured as glucose with a commercially available glucose C-II test Wako (manufactured by Wako Pure Chemical Industries). The chain length distribution of the α-glucan contained was evaluated using an MCI-gel CK02AS column (20 × 250 mm) and a corona CAD detector without performing glucoamylase treatment. Specifically, the degree of polymerization (chain length) was analyzed under the following conditions. G1-G7 malto-oligosaccharide and amylose DP17 were used as mobility standards.

Column: MCI-gel CK02AS column (20 × 250 mm, manufactured by Mitsubishi Chemical Corporation)
Mobile phase: MQ water flow rate: 1.0 mL / min
Column temperature: 85 ° C
Detector: Corona CAD

(6) Sensory evaluation These fractionated / purified samples are added to a commercial beer-based alcoholic beverage using barley and barley malt with a malt use ratio of less than 49% and 50% of each fraction contained in the beverage. (See FIG. 1 and FIG. 2), sensory evaluation was performed by a panel of five people.

  The index of sensory evaluation was evaluated with a five-point score of 1 to 5 points as a comprehensive evaluation of “umami, sweetness, thickness, body, and astringency / taste dissonance as an off-flavor”. The additive-free control was scored 2.5. The average value of the sensory evaluation score is shown in FIG. 3, and the sensory evaluation comments are shown in Table 3.

  The peptide fraction had a high sensory evaluation score (Fig. 3) in test group 2 from the pre-fraction to D, which was consistent with the sensory evaluation result of the fermentation broth before fractionation. Moreover, in the fractions A1 and A2 of the test group 2, the score was high, and it was a sensory evaluation comment that softness and miscellaneous taste decreased (Table 3). In the pre-fraction of Test Zone 2, the score was equally high, but it was a sensory evaluation comment that it was soft but had less taste. Fractions B to D were equally high in score, but sensory evaluation comments were evaluated to have a greater contribution of thickness, body, and umami. That is, in the pre-fractions, A1, A2, B, C, and D, the evaluation of the test section 2 is high, but the taste qualities are different, and the fractions A1 and A2 are beer-like softness, poor taste, etc. It turned out that there is an effect.

  In the α-glucan fraction, when fractions B and C were added, the sensory evaluation score (FIG. 3) was high, and the sensory evaluation comment (Table 3) was expressed as sweet, mellow, and smooth. . The α-glucan derived from fractions A1 and A2 did not have a clear flavor.

  The peptide fraction was analyzed with a Superdex 75 10/300 column described in Example 2 and the molecular weight was estimated. As a result, the peptide of fraction A1 was distributed in a molecular weight of about 10 to 20 kDa, and SDS-PAGE Also on electrophoresis, it was confirmed that similar peptides having a molecular weight of about 10 to 20 kDa were distributed in the fractions A1 to A2 (data not shown). Moreover, it was confirmed that the amount of the peptide increased in the test group 2 which was excellent in flavor (FIG. 1).

  Moreover, it was as Table 4 when the polymerization degree distribution of the alpha-glucan fraction was confirmed.

  As shown in Table 4, it was confirmed that the main components of fractions B and C in which a clear flavor was felt included a degree of polymerization (DP) in the range of 2 to 10. Moreover, in the test group 2 which was excellent in flavor, it was confirmed that the amount of α-glucan in the fraction was increased. In the α-glucan derived from fractions A1 and A2, the chain length of the main component was longer (8 to about 40), and no clear flavor was felt.

  From the above results, the fractionated and purified peptide fractions A1 and A2 having a molecular weight of about 10 to 20 kDa and α-glucan fractions B and C having a degree of polymerization of glucose of 2 to 10 are suppressed and harmonious. It became clear that a beer-like taste can be imparted to beer-based beverages.

Example 2: Component analysis and sensory evaluation of beer-taste alcoholic beverage (1) Production of beer-taste alcoholic beverage A beer-taste alcoholic beverage was produced at a pilot plant. The malt ratio in the raw materials used for the beer-taste alcoholic beverage was less than 50% and less than 25%.

  In the production of beer-taste alcoholic beverages, barley malt was used as a main ingredient, and any one or more of barley, corn grits, corn starch, rice, liquid sugar was used as an auxiliary ingredient. In the saccharification, an enzyme preparation was used, and the temperature and time of saccharification were adjusted and filtered to obtain worts having different compositions. That is, the temperature range of saccharification was selected in 50-65 degreeC, such as 50, 60, or 65 degreeC. The saccharification time was adjusted between 5 and 140 minutes at each temperature step. The heat treatment temperature of the raw material was selected from 70 to 100 ° C. Specifically, wort was obtained as follows.

(A) Sample No. Saccharification conditions of 16 and 17 For 100 parts by mass of hot water at 60 ° C., 27.1 parts by mass of corn grits and 2.1 parts by mass of barley malt were added and held for 5 minutes, then heated to 78 ° C. and held for 5 minutes. Further, the temperature was raised to 100 ° C. and held for 30 minutes was designated as A 醪. In another kettle, 27.1 parts by weight of barley malt, 45.0 parts by weight of barley and an enzyme preparation were put into 246.4 parts by weight of hot water at 50 ° C. The obtained A 醪 and B 醪 were immediately mixed to 65 ° C. Then, after hold | maintaining at 65 degreeC for 100 to 140 minutes, after heating up at 78 degreeC and hold | maintaining for 5 minutes, it filtered and wort was obtained.

(A) Sample No. 7 saccharification conditions 1100 parts by weight of barley malt, 18.7 parts by weight of barley, 100 parts by weight of hot water at 50 ° C., 70 minutes after holding the enzyme preparation and heating to 65 ° C. and holding for 130 minutes The temperature was further raised to 78 ° C. and held for 5 minutes, followed by filtration to obtain wort.

(C) Sample No. 8-10 saccharification conditions For 100 parts by mass of hot water at 50 ° C., 30.4 parts by mass of barley, 3.0 parts by mass of barley malt, and the enzyme preparation were held for 60 minutes, then heated to 60 ° C. and 20 The sample was held for 40 minutes, further heated to 70 to 80 ° C. and held for 40 minutes, which was designated as A 醪. In another kettle, 62.5 parts by mass of hot water at 50 ° C. and 34.2 parts by mass of barley malt and the enzyme preparation were added and held for 20 minutes to make B koji. The obtained A 醪 and B 醪 were immediately mixed, and the mixture was held at 65 ° C. for 100 minutes as an AB mixing vessel, heated to 78 ° C. and held for 5 minutes, and then filtered to obtain wort.

(D) Sample No. 14 saccharification conditions For 100 parts by weight of hot water at 50 ° C., 6.8 parts by weight of barley malt, 26.3 parts by weight of barley, an enzyme preparation were held for 60 minutes, then heated to 65 ° C. and held for 100 minutes The temperature was further raised to 78 ° C. and held for 10 minutes, followed by filtration to obtain wort.

(E) Sample No. 15 saccharification conditions For 100 parts by mass of hot water at 50 ° C., 30.4 parts by mass of corn grits and 2.8 parts by mass of barley malt were added, held for 5 minutes, then heated to 78 ° C. and held for 5 minutes. What was heated up to 100 degreeC and hold | maintained for 40 minutes was made into A *. In a separate kettle, 4.8 parts by weight of barley malt, 50.4 parts by weight of barley, and enzyme preparation were added to 148.0 parts by weight of hot water at 50 ° C., held for 60 minutes, and then heated to 70 ° C. What was hold | maintained for 30 minutes was made into B *. Further, in another kettle, 20.2 parts by mass of barley malt and enzyme preparation were added to 140.0 parts by mass of hot water at 50 ° C. and held for 30 minutes to make C koji. The obtained B 醪 and C 醪 were immediately mixed and held at 60 ° C. for 15 minutes to obtain D 醪. The obtained A 醪 and D 醪 were immediately mixed and held at 65 ° C. for 80 minutes, further heated to 78 ° C. and held for 10 minutes, and then filtered to obtain wort.

(F) Sample No. 18 saccharification conditions For 100 parts by mass of hot water at 60 ° C., 27.1 parts by mass of corn grits and 2.5 parts by mass of barley malt were added, held for 10 minutes, heated to 100 ° C. and held for 30 minutes. A. In another kettle, 15.7 parts by weight of barley malt, 45.0 parts by weight of barley and enzyme preparation were put into 246.4 parts by weight of hot water at 50 ° C. and held for 45 minutes. B parts were prepared by adding 25 parts by weight of the enzyme preparation and the enzyme preparation. The obtained A 醪 and B 醪 were immediately mixed to 65 ° C. Then, after hold | maintaining at 65 degreeC for 100 minutes, after heating up to 78 degreeC and hold | maintaining for 5 minutes, it filtered and wort was obtained.

  Following the above wort preparation steps (a) to (f), hops and some worts (samples Nos. 15 to 18) are further added to each wort obtained to add liquid sugar to 100 ° C. After boiling for 60 to 70 minutes, the wort was left standing to separate the trube, and then cooled to obtain a pre-fermentation solution. Thereafter, the bottom fermentation yeast was added to the pre-fermentation solution to prepare a fermentation solution. Main fermentation was performed by maintaining this fermentation broth at a predetermined temperature for a predetermined period. Furthermore, post-fermentation was performed by maintaining the fermentation liquid after the main fermentation at a predetermined temperature for a predetermined period. Subsequently, the fermented liquid after post-fermentation was stored by maintaining it at a lower temperature for a predetermined period and filtered to obtain clear beer-taste alcoholic beverages (samples Nos. 7 to 10 and 14 to 18).

  In addition, a commercial product (beer) having a malt use ratio of 67% or more as samples 1 to 3, a commercial product (malt beer) having a malt use ratio of 50% or less as samples 4 to 6, and a commercial product having a malt use ratio of 25% or less. (New genre-based alcoholic beverages) were subjected to analytical tests as Samples 11 to 13 and 19, respectively.

(2) Analysis of amount of α-glucan having a degree of polymerization of 5 to 10 (a) Preparation of sample for saccharide analysis Beer product, product sample produced in test brewing equipment, weighed, 30% (v / v) acetonitrile / 70% (V / v) A sample diluted 50 times with an aqueous solution was used as a sample for quantification.

(A) Determination of sugars by LC / MS / MS HPLC conditions were as shown in Table 5.
Equipment used: 1200 Series (Agilent technologies)
Column: ACQUITY UPLC BEH Amide 2.1mm x 50mm (particle size 1.7μm) (Waters)
Mobile phase A: 95% (v / v) acetonitrile water + 0.1% (v / v) formic acid + 10 mM ammonium formate Mobile phase B: Water + 0.1% (v / v) formic acid + 10 mM ammonium formate Injection amount: 5 μL
Column temperature: 55 ° C
Sample temperature: 20 ° C
Gradient conditions / flow velocity:

(C) Setting mass spectrometer conditions For Maltopentaose, Maltohexaose, and Maltoheptaose, purchase standard products from Tokyo Kasei Co., Ltd., Maltooctaose, and Maltonona. Standard materials were purchased from Elicityl SA for Maltononaose and Maltodecaose, and mass spectrometer conditions were set. The purchased standard was adjusted to 100 ppm using 50% (v / v) acetonitrile / 50% (v / v) water / 0.1% (v / v) formic acid / 10 mM ammonium formate solution, and 3200Q trap manufactured by ABSciex. The various parameters were optimized by direct injection.

(D) Analysis of sample The actual sample was analyzed under the analysis conditions set in (c) above. Five samples obtained by adding standard samples with different concentrations to the matrix were analyzed, and a calibration curve was created from the results by linear approximation using the least square method, and quantification was performed using only the slope. For each sample, a sample in which a standard product with a known concentration was added in addition to the sample for quantification was analyzed, and it was confirmed that the recovery rate was 75% or more and 135% or less. The results were as shown in Table 6.

  In the α-glucan addition test of Example 1, the strong flavor imparting effect was confirmed to be in the range of the polymerization degree of 2 to 10, and the correlation between the component strength and the flavor of various polymerization degrees was examined. As a result, since it was confirmed that the correlation was high (data omitted), the concentration of α-glucan (maltooligosaccharide) having a polymerization degree of 5 to 10 was set as a component index.

(3) Analysis of 10-20 kDa peptide amount (a) Sample preparation for gel filtration fractionation Product samples and commercial products produced in a pilot plant were weighed and freeze-dried. The dried product was dissolved in 50 mM phosphate buffer (containing 150 mM NaCl) to prepare a 2.5-fold concentrated solution, which was used as a fractionation sample.

(A) HPLC gel filtration fraction The conditions of HPLC gel filtration fraction were as follows.
<HPLC gel filtration fractionation conditions>
Column: Superdex 75 10/300 (manufactured by GE Healthcare)
Sample injection volume: 100 μL
Eluent composition: 50 mM phosphoric acid, 20% (v / v) acetonitrile, 150 mM NaCl
Flow rate: 0.5 mL / min (constant flow rate)
Detection wavelength: 215 nm
Fractionation program:

(C) Setting of the fraction range What is obtained by appropriately dissolving a peptide having a known molecular weight in ultrapure water at 0.1 to 5 mg / mL in the column, eluent, flow rate, and detection wavelength described in the above HPLC gel filtration fraction conditions. A 50 μL injection was performed and HPLC analysis was performed to confirm the retention time (Table 8). A calibration curve (FIG. 4) was created from the retention time and molecular weight, and the molecular weight range and fractional range were determined.

(D) Protein quantification of the fraction by the Lowry method Protein quantification was performed by the Lowry method described in Example 1. A calibration curve was prepared from the obtained absorbance and BSA concentration, the peptide amount of the fraction solution (BSA conversion) was calculated, and the product equivalent peptide concentration (mg / mg) of the fraction was calculated from the fraction solution amount and the concentration ratio. ml) was calculated.

(E) Quantification of 10-20 kDa peptide The 10-20 kDa peptide concentration is the product equivalent peptide of the protein concentration contained in fraction 3 in the molecular weight range of 10-20 kDa determined from the calibration curve in the HPLC gel filtration fraction. It was calculated in terms of concentration (mg / ml). Further, the ratio of the 10-20 kDa peptide amount to the total protein amount, that is, the 10-20 kDa peptide ratio was determined by the following calculation formula.
Peptide ratio (%) of 10-20 kDa = 10-20 kDa peptide amount (mg / product ml) / total protein amount (mg / product ml) × 100

The results were as shown in Table 9.

(4) Sensory evaluation of beer-taste alcoholic beverages The beer-taste alcoholic beverages obtained by manufacturing were subjected to sensory evaluations by six trained panelists. Moreover, sensory evaluation was carried out by eight trained panelists regarding commercially available beer-taste alcoholic beverages. The evaluation items were as follows.

  As evaluation item 1, sensory evaluation was performed on 9 levels from 1 (not like beer) to 9 (like beer) for beer-like taste (soft and smooth texture as in beer is felt and harmonious). In addition, as evaluation item 2, sensory evaluation was performed on 9 roughnesses ranging from 1 (weak) to 9 (strong) for roughness remaining in the mouth (a bitter taste such as astringency and a rough feel remaining on the tongue). The results of component analysis and sensory evaluation of each sample are as shown in Table 10.

  As shown in Table 10, sample No. which is a commercial beer. 1 to 3, the beer-like taste was as high as 7.1 to 7.3, and the roughness remaining in the mouth was as low as 3.1 to 4.5. In addition, when the malt ratio is less than 50% and the malt ratio is less than 25%, the peptide concentration of 10-20 kDa is 0.19 mg / mL product or more, and the peptide ratio of 10-20 kDa is 3.7% or more, In addition, when the concentration of α-glucan having a polymerization degree of 5 to 10 is 3.3 mg / ml or more (specifically, in the case of sample Nos. 7, 8, 10, 14, and 18), the taste like beer is obtained. The roughness increased to 5.2 to 6.6, the roughness remaining in the mouth decreased to 3.8 to 5.1, and a tendency to approach the beer score was observed.

  Moreover, the bubble graph of the sensory evaluation result of the taste like beer was shown in FIG. 5 and FIG. In any figure, it turned out that the sensory evaluation score of the taste like beer becomes 5.1 or more in the sample plotted in the range surrounded by the frame.

  From the above results, when the beer-taste alcoholic beverage contains a peptide concentration of 10 to 20 kDa and the ratio thereof, and the total concentration of α-glucan having a polymerization degree of 5 to 10 is included in a specific range, such as in beer It was shown that a soft and smooth texture was felt, there were few astringent tastes such as astringency and roughness, and the sense of harmony was improved.

Example 3: Evaluation of relationship between α-glucan, peptide and sensory evaluation For beer-taste sparkling alcoholic beverages such as commercially available beer, happoshu, and new genre beverages, α-glucan concentration of 5 to 10 and 10 to 20 kDa The correlation coefficient (R-square) between the peptide concentration and sensory evaluation was evaluated. The analysis method for the α-glucan concentration of 5 to 10 and the peptide concentration of 10 to 20 kDa is as shown in Example 2. The numerical value including both of the two components was calculated by multiplying the α-glucan concentration (mg / ml) having a degree of polymerization of 5 to 10 and the peptide concentration (mg / ml) of 10 to 20 kDa. The sensory evaluation was performed by 10 trained panelists, with “taste harmony”, that is, harmony of taste related to beer-like, as an index. It was evaluated as having never felt in the person's recognition.

  Table 11 shows the α-glucan concentration with a degree of polymerization of 5 to 10, the peptide concentration of 10 to 20 kDa, and the sensory evaluation results. Further, the relationship between the concentration and the sensory evaluation result was as shown in FIG.

  As shown in Table 11 and FIG. 7, the correlation coefficient with the sensory evaluation result was obtained for each of the α-glucan concentration at a polymerization degree of 5 to 10 and the peptide concentration of 10 to 20 kDa (R2 = 0 respectively). .29, 0.46), the correlation coefficient between the two components (the value obtained by multiplying the concentrations of the two components) is higher (R2 = 0.58). From this result, it was confirmed that the α-glucan having a degree of polymerization of 5 to 10 and the peptide having a degree of polymerization of 10 to 20 kDa were not alone, but the effectiveness of using two components as indexes.

Example 4: Addition of a peptide fraction and an α-glucan fraction to a beer-taste alcoholic beverage and sensory evaluation (part 2)
(1) Production of beer-taste fermented alcoholic beverage and preparation and analysis of fractionated purified product Sample 1 described in Example 2 (commercial beer product of malt 67% or more) and sample in the same manner as described in Example 1 Peptide fractions (pre, A1, A2, B, C) and α-glucan fractions (B, C) were prepared from 7 (test brewed product with less than 50% malt). The determination of the amount of 10-20 kDa peptide and the analysis of the amount of α-glucan (maltooligosaccharide) having a degree of polymerization of 5-10 were also carried out in the same manner as described in Example 2. The prepared fractionated / purified samples were added to the following commercial products and test brews so as to have the amounts shown in Table 12, and sensory evaluation was performed by a panel of six persons. In addition, purified fraction pres, A1 + A2 and B + C were added to sample no. 4 was added so that it might become the quantity shown in Table 13, and sensory evaluation was similarly performed by the panel of six persons.

(2) Sensory evaluation The items of sensory evaluation were the same as in Example 2. That is, as evaluation item 1, taste like beer (a soft and smooth texture as in beer is felt and harmonious) is categorized in 9 levels from 1 (not beer) to 9 (beer) evaluated. In addition, as evaluation item 2, sensory evaluation of roughness remaining in the mouth (miscellaneous taste such as astringency and a rough feel remaining on the tongue) was performed in 9 stages from 1 point (weak) to 9 points (strong). The sensory results and analysis results were as shown in Table 12 and FIGS.

  From these results, when the amount of α-glucan having a 10-20 kDa peptide or a polymerization degree of 5-10 increases to a certain value or more, the sensory evaluation score improves, and when both of these amounts exceed a certain value, It became clear that evaluation score improved more effectively. Moreover, even when it did not reach a fixed value, it became clear that the sensory evaluation score tends to improve when the amount of α-glucan having a 10-20 kDa peptide or a polymerization degree of 5-10 increases.

  When combined with the data of Example 2, when the malt ratio is less than 50% and the malt ratio is less than 25%, the peptide concentration of 10-20 kDa is 0.15 mg / mL or more and the peptide ratio of 10-20 kDa is When the concentration of α-glucan having a degree of polymerization of 5 to 10 is 3.3 mg / mL or more, the taste like beer is as high as 5.1 or more, and the roughness remaining in the mouth is 5 It became clear that the sensory evaluation score tended to improve.

  In addition, when the concentration of α-glucan having a polymerization degree of 5 to 10 is 2.1 mg / mL or more, the taste like beer is 4.2 or more, the roughness remaining in the mouth is 5.1 or less, and the sensory evaluation score Tended to be close to beer scores.

  Further, as shown in Table 13 and FIG. 10, the purified fraction pres, A1 + A2 and B + C were used as sample Nos. When added to No. 4, it was revealed that fraction A1 + A2 had the highest beer-like taste score and reduced roughness.

Example 5: Identification of molecular species of protein fraction In this example, an attempt was made to identify a protein that contributes to a beer-like taste by 2D-PAGE and MALDI-TOF-MS.

  200 μL of an equal volume mixture of fractions A1 and A2 of the gel filtration fraction obtained in the same manner as in Example 1 was purified by TCA acetone precipitation, followed by 2D-PAGE electrophoresis and silver staining. . FIG. 11 shows the result of 2D-PAGE. FIG. 11A is a prescription for test section 1 of Example 1, and FIG. 11B is a prescription for test section 2 of Example 1. Moreover, FIG. 11C is the sample 16 of Example 2, and FIG. 11D is the sample 15 of Example 2. FIG. As can be seen from FIGS. 11A and 11B, it was found that there was a quantitative difference between the samples in the test group 1 and the test group 2 in the band in the circled range. Further, as can be seen from C and D in FIG. 11, it was found that there is a quantitative difference between the sample 16 and the sample 15 in the band in the circled range.

  Next, the band indicated by the arrow shown in FIG. 11 was cut out, and the protein was digested using trypsin (In-Solution Tryptic Digestion and Guanidination Kit, manufactured by Thermo Scientific). The obtained trypsin-digested peptide solution was desalted and concentrated using a resin-filled pipette tip (Zip-Tip C18, manufactured by Merck Millipore). The desalted and concentrated peptide solution and the matrix (10 μg / μL α-cyano-4-hydroxycinnamic acid / methanol solution) were mixed on the plate in equal amounts, and then MS was prepared using MALDI-TOF-MS (manufactured by Bruker Daltonics). Spectra were acquired and proteins were identified by Mascot / SWISS-PROT database search.

The results of protein identification by MALDI-TOF-MS were as shown in Table 14.

Examples of the protein having a difference in amount in the gel filtration fraction (fractions A1 to A2) having a difference in flavor between the test sections 1 and 2 of Example 1 include α-amylase / trypsin inhibitor CMb, α-amylase. / Trypsin inhibitor CMd and trypsin inhibitor Cme were identified. Examples of proteins having a difference in gel filtration fraction (fractions A1 to A2) between sample 16 and sample 15 of Example 2 include α-amylase / trypsin inhibitor CMb and α-amylase / trypsin inhibitor CMd. , Trypsin inhibitor Cme, serpin-Z4, non-specific lipid-transfer protein 1 were identified. The identified proteins are all derived from barley ( Hordeum vulgare ). α-amylase / trypsin inhibitor CMb, α-amylase / trypsin inhibitor CMd, trypsin inhibitor Cme, serpin-Z4 are known as protease inhibitor proteins, and non-specific lipid-transfer protein 1 is known as a lipid transfer protein. ing.

Claims (20)

  A beer-taste fermentation alcoholic beverage comprising malt and / or ungerminated wheat as a raw material, wherein the peptide concentration of molecular weight 10-20 kDa (gel filtration method) is 0.15 mg / ml or more. Alcoholic beverages.   The beer-taste fermented alcoholic beverage according to claim 1, wherein the malt use ratio is less than two-thirds.   The beer-taste fermented alcoholic beverage according to claim 1 or 2, wherein a ratio of a peptide amount having a molecular weight of 10 to 20 kDa to a total protein amount is larger than 3.6%.   The beer-taste fermented alcoholic beverage according to any one of claims 1 to 3, wherein a peptide concentration of a molecular weight of 10 to 20 kDa is 0.19 mg / ml or more. The beer-taste fermented alcoholic beverage according to any one of claims 1 to 4, wherein the concentration of α-glucan having a polymerization degree of 5 to 10 is 2.1 mg / ml or more. The beer-taste fermented alcoholic beverage according to claim 5, wherein the concentration of α-glucan having a polymerization degree of 5 to 10 is 3.3 mg / ml or more.   A beer taste comprising one or more peptides having a molecular weight of 10 to 20 kDa (gel filtration method) derived from a beer-taste fermented alcoholic beverage made of malt and / or ungerminated wheat as a raw material. Alcoholic beverages.   The beer-taste alcoholic beverage of Claim 7 mix | blended so that it may become a density | concentration of 0.15 mg / ml or more of peptides with a molecular weight of 10-20 kDa.   The beer-taste alcoholic beverage of Claim 7 or 8 mix | blended so that a peptide with a molecular weight of 10-20 kDa may become a ratio larger than 3.6% with respect to the total protein amount in a drink.   8. One or more kinds of α-glucan having a degree of polymerization of 5 to 10 derived from a beer-taste fermented alcoholic beverage that uses malt and / or ungerminated wheat as a raw material are further blended. The beer-taste alcoholic drink as described in any one of -9.   The beer-taste alcoholic beverage of Claim 10 mix | blended so that it may become a density | concentration of 2.1 mg / ml or more of alpha-glucan with a polymerization degree of 5-10.   One or more peptides having a molecular weight of 10-20 kDa are selected from the group consisting of α-amylase / trypsin inhibitor, serpin-Z4 and non-specific lipid-transfer protein 1 The beer-taste alcoholic beverage as described in any one of Claims 7-11 which is 1 type, or 2 or more types.   Comprising a step of blending one or more peptides having a molecular weight of 10 to 20 kDa (gel filtration method) derived from a beer-taste fermented alcoholic beverage that uses malt and / or ungerminated wheat as a raw material. The manufacturing method of a beer taste alcoholic beverage.   The manufacturing method of the beer taste alcoholic beverage of Claim 13 mix | blended so that it may become a density | concentration of 0.15 mg / ml or more of peptides with a molecular weight of 10-20 kDa.   The manufacturing method of the beer taste alcoholic beverage of Claim 13 or 14 mix | blended so that a peptide with a molecular weight of 10-20 kDa may become a ratio larger than 3.6% with respect to the total protein amount in a drink.   A step of further blending one or two or more α-glucans having a degree of polymerization of 5 to 10 derived from a beer-taste fermented alcoholic beverage that uses malt and / or ungerminated wheat as a raw material, The manufacturing method of the beer taste alcoholic beverage as described in any one of Claims 13-15.   The manufacturing method of the beer taste alcoholic beverage of Claim 16 mix | blended so that the alpha-glucan with a polymerization degree of 5-10 may become a density | concentration of 2.1 mg / ml or more.   One or more peptides having a molecular weight of 10-20 kDa are selected from the group consisting of α-amylase / trypsin inhibitor, serpin-Z4 and non-specific lipid-transfer protein 1 The manufacturing method of the beer taste alcoholic beverage as described in any one of Claims 13-17 which is 1 type, or 2 or more types.   Comprising, as an active ingredient, one or more peptides having a molecular weight of 10 to 20 kDa (gel filtration method) derived from beer-taste fermented alcoholic beverages that use malt and / or ungerminated wheat as part of the raw material, A flavor improver for beer-taste alcoholic beverages. A step of adding one or two or more peptides having a molecular weight of 10 to 20 kDa (gel filtration method) derived from a beer-taste fermented alcoholic beverage made of malt and / or ungerminated wheat as a raw material. , How to improve the flavor of beer-taste alcoholic beverages.