JP2017123837A - Agent for improving astringency and mildness of beer taste beverage, and method for improving astringency and mildness of beer taste beverage - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide means capable of reducing astringency of a beer taste beverage and means capable of improving mildness thereof.SOLUTION: The present invention provides an agent for improving astringency and mildness of a beer taste beverage that comprises lipoxygenase-1 deleted barley material as an active ingredient.SELECTED DRAWING: None

Description

  The present invention relates to a beer-taste beverage astringency improving agent and mildness improving agent, and a beer-taste beverage astringency improving method and mildness improving method.

  In the production of beer-taste beverages, various techniques have been proposed to improve the flavor. For example, Patent Document 1 includes a step of adding α-glucosidase in order to produce a low-alcohol beer-taste beverage having sufficient richness and flavor and high drinkability, and the amount of extract, the degree of fermentation, etc. Is disclosed to be adjusted within a certain range. Patent Document 2 discloses a method including a step of fermenting a raw material of an alcoholic beverage with yeast in the presence of a substance belonging to the vitamin B group in order to obtain an alcoholic beverage having a low indole concentration.

Japanese Patent Laying-Open No. 2015-133924 JP 2009-278921 A JP 2008-43348 A JP 2005-102690 A

  In beer-taste beverages, there is still room for improvement in terms of flavor. An object of this invention is to provide the means which can reduce the astringency of a beer taste drink. Moreover, an object of this invention is to provide the means which can improve the mildness of a beer taste drink.

  The present invention provides a beer-taste beverage astringency improving agent and a mildness improving agent comprising a lipoxygenase-1-deficient barley material as an active ingredient.

  The present invention also provides a method for improving the astringency and mildness of beer-taste beverages, comprising using a lipoxygenase-1 deficient barley material as a barley material.

  In the method for improving the astringency of beer-taste beverages, the ratio of the lipoxygenase-1-deficient barley material in the total barley material is preferably 20% or more. When the ratio of the lipoxygenase-1 deficient barley material is within the above range, the astringency of the beer-taste beverage can be more efficiently reduced.

  In the method for improving the mildness of a beer-taste beverage, the ratio of the lipoxygenase-1-deficient barley material in the total barley material is preferably 10% or more. When the ratio of the lipoxygenase-1 deficient barley material is within the above range, the mildness of the beer-taste beverage can be improved more efficiently.

  In the astringency improving method and the mildness improving method, the barley raw material may be malt, and the method may include a step of obtaining wort from the malt.

  With the astringency improving agent or the astringency improving method of the present invention, the astringency of a beer-taste beverage can be reduced. Moreover, the mildness of a beer taste drink can be improved by the mildness improving agent or mildness improving method of the present invention.

  Hereinafter, embodiments for carrying out the present invention will be described in detail. In addition, this invention is not limited to the following embodiment.

  The astringency improving agent and mildness improving agent for beer-taste beverages of the present invention contain a lipoxygenase-1-deficient barley material as an active ingredient.

  Lipoxygenase-1 (hereinafter referred to as “LOX-1”), an enzyme contained in barley, oxidizes linoleic acid and produces 9-hydroperoxyoctadecadienoic acid in the manufacturing process of beverages containing barley ingredients as raw materials. It is known to generate.

  In the present specification, the barley raw material refers to barley or processed barley. Examples of processed barley products include barley extract, malt, wort, and malt extract.

  In this specification, the LOX-1 deficient barley material is a barley material lacking LOX-1. In the present specification, LOX-1 deletion means substantially not having LOX-1 protein, or having LOX-1 protein but having substantially no LOX-1 activity. That means. The LOX-1 deficient barley material can be selected, for example, by measuring and confirming the expression level of LOX-1 protein or LOX-1 activity. The LOX-1 deficient barley material only needs to lack LOX-1 when used in the production of beer-taste beverages. The improving agent according to the present embodiment can reduce the astringency of a beer-taste beverage and improve mildness by including a barley raw material lacking LOX-1 as an active ingredient. it can.

  As the LOX-1 deficient barley raw material, for example, barley lacking LOX-1 (hereinafter also referred to as “LOX-1 deficient barley”) is used as barley in a known barley raw material production method. Can be obtained. In this case, since the LOX-1 deficient barley raw material can be obtained simply by replacing barley with LOX-1 deficient barley, it is convenient.

  LOX-1 deficient barley can be selected, for example, by measuring and confirming the expression level of LOX-1 protein or LOX-1 activity. Moreover, you may use the barley which has the barley LOX-1 mutant gene mentioned later as LOX-1 deletion barley.

The LOX-1 activity of barley can be confirmed, for example, by measuring the oxidative activity of linoleic acid in a crude enzyme solution extracted from barley seeds or malt. Specifically, for example, it can be measured by the following method.
One ripe barley seed is crushed and extracted with 500 μL of extraction buffer (0.1 M sodium acetate buffer (pH 5.5)) at 4 ° C. for 30 minutes. The obtained extract is centrifuged at 15000 rpm for 10 minutes, and then the supernatant is obtained as a crude enzyme solution. Next, 5 μL of a substrate solution (40 mM linoleic acid, 1.0% (W / V) Tween 20 aqueous solution) and 85 μL of extraction buffer are added to 10 μL of the crude enzyme solution and mixed, and then reacted at 24 ° C. for 5 minutes. The reaction is stopped by adding and mixing 100 μL of a reaction stop solution (80 mM 2,6-di-t-butyl-p-cresol, methanol solution). The reaction solution is allowed to stand at −20 ° C. for 30 minutes, and then centrifuged at 3000 rpm for 20 minutes to obtain a supernatant. To 20 μL of the obtained supernatant, 200 μL of color developing solution (4 mM 2,6-di-t-butyl-p-cresol, 25 mM sulfuric acid, 0.25 mM ammonium iron (II) sulfate hexahydrate, 100 mM xylenol orange, 90% (Methanol aqueous solution) is added, and after standing for 30 minutes, the absorbance at a wavelength of 550 nm is measured. As a negative control, a crude enzyme solution was heat-treated at 100 ° C. for 5 minutes, and a reaction in which LOX-1 was inactivated in the same manner was used. As a positive control, a crude enzyme solution of seeds of barley variety Kendall was used. .

The expression level of barley LOX-1 protein can be examined by Western blotting using, for example, an anti-LOX-1 antibody. Specifically, for example, the following method can be used.
3 μg of total soluble protein extracted from barley using 0.1 M sodium acetate buffer (pH 5.5) is fractionated by SDS polyacrylamide electrophoresis (SDS-PAGE) and then blotted onto a PVDF membrane (Millipore). After washing this membrane with TTBS (20 mM Tris-HCl (pH 7.5), 0.15 M sodium chloride, 0.05% (w / v) Tween 20, 0.05% (w / v) sodium azide)), React with the LOX-1 antibody solution (1000-fold dilution / TTBS) for 30 minutes. The membrane is washed with TTBS for 5 minutes three times, and then reacted with an alkaline phosphatase-labeled goat anti-rabbit IgG antibody solution (Santa Cruz, 1000-fold diluted TTBS solution) for 30 minutes. The membrane was washed with TTBS for 5 minutes × 2 times, and further washed with AP9.5 (10 mM Tris-HCl (pH 9.5), 0.1 M sodium chloride, 5 mM magnesium chloride) for 5 minutes × 1 time. Color is developed by reacting with an alkaline phosphatase substrate solution (1 mg / ml nitro blue tetrazolium, 0.5 mg / ml BCIP, AP 9.5 solution). Thereby, the expression level can be compared according to the strength of the band relative to the subject.

  LOX-1-deficient barley is, for example, barley with a barley LOX-1 mutant gene in which the guanine of the splicing donor site (5′-GT-3 ′) of the fifth intron of barley LOX-1 gene is mutated to adenine. is there. The mutant gene is characterized in that the 60th base G of the fifth intron of the LOX-1 gene is replaced with A as compared to the known LOX-1 gene (SEQ ID NO: 1) (SEQ ID NO: 2). . Since the 60th-61st position of the 5th intron of the LOX-1 gene is a splicing donor site (5′-GT-3 ′), substitution of this base causes LOX-1 to be spliced abnormally and active LOX-1 Cannot be expressed. In the sequence listing, SEQ ID NO: 1 shows the nucleotide sequence of the fifth intron region of the known LOX-1 gene, and SEQ ID NO: 2 shows the nucleotide sequence of the fifth intron region of the LOX-1 mutant gene of the present invention. The base sequence of the corresponding part is shown. The above-mentioned LOX-1 activity measurement method, LOX-1 protein expression level measurement method, LOX-1 deficient barley having a barley LOX-1 mutant gene, LOX-1 deficient barley selection method and the like are described in detail in Patent Document 3. Have been described.

  As described above, LOX-1 deficient barley has the characteristics of not expressing LOX-1 protein or having substantially no LOX-1 activity as described above, and is suitable for production of beer-taste beverages. It is preferable to have characteristics of varieties, characteristics of barley varieties suitable for breeding, and the like. LOX-1 deficient barley having desired characteristics can be obtained, for example, by crossing LOX-1 deficient barley having a LOX-1 mutant gene with a variety having the characteristics.

  The LOX-1 deficient barley raw material can also be obtained by using conventional barley, that is, barley that does not lack LOX-1, by making it by a known method for reducing LOX-1. (For example, refer to Patent Document 4).

  In this specification, a beer-taste beverage refers to a beverage that exhibits a taste and aroma like beer and gives a drinker a sense of drinking beer during drinking. The beer-taste beverage may be an alcoholic beverage or a non-alcoholic beverage. Examples of beer-taste alcoholic beverages include beer, happoshu, other brewed liquors (foaming (1)), and liqueurs under the Japan Sake Tax Act (Act No. 6 of February 28, 1948). The thing classified into (foaming property (1)) is mentioned. Non-alcohol means that alcohol is not substantially contained. The alcohol content of the non-alcohol beer-taste beverage is, for example, 0.00%. In the present specification, alcohol means ethanol unless otherwise specified.

  A beer taste drink can be manufactured by the same method as the manufacturing method of a well-known beer taste drink except using the astringency improving agent or mildness improving agent which concerns on this embodiment as at least one part of a raw material. That is, a beer-taste beverage with improved astringency and / or mildness can be obtained by replacing part or all of the raw materials used in the production of a known beer-taste beverage with the improver according to this embodiment. . The improving agent according to the present embodiment can be used for producing a beer-taste beverage in the same manner as a normal barley raw material without undergoing a special treatment. Therefore, by using the improving agent according to the present embodiment, for example, it is not necessary to perform fine control such as fermentation conditions or adjustment of trace components in the beverage, and the astringency and / or mildness of the beer-taste beverage can be easily achieved. Can be improved.

  The present invention also provides a method for improving the astringency of beer-taste beverages and a method for improving mildness, including the use of LOX-1 deficient barley materials as barley materials. In the manufacturing process of a beer-taste beverage, when the barley raw material contains the LOX-1 deficient barley raw material, the astringency of the resulting beer-taste beverage can be reduced and the mildness can be improved.

  In the above improvement method, a beer-taste beverage can be obtained by the same method as a known beer-taste beverage production method, except that the LOX-1 deficient barley material is used as at least part of the barley material. Astringency and / or mildness can be easily improved. Examples of the use of the LOX-1 deficient barley material include a step of obtaining a LOX-1 deficient malt from LOX-1 deficient barley, and a LOX-1 deficient malt from barley not lacking LOX-1. Step, Step of obtaining LOX-1 deficient wort from LOX-1 deficient malt, step of fermenting LOX-1 deficient wort, step of adding processed LOX-1 deficient barley to beer-taste beverage ingredients, etc. Is mentioned.

  Aspects such as barley raw materials, LOX-1 deficient barley, and beer-taste beverages in the astringency improving method and mildness improving method of beer-taste beverages can apply the same aspects as those described in the above-described improving agent.

  The barley material is preferably malt. When the barley raw material is malt, the improvement method may include a step of obtaining wort from the malt.

  When the beer-taste beverage is beer, for example, after a step of obtaining wort from malt, a step of boiling wort, a step of adding hops to wort before or after boiling, agglomerates in a whirlpool tank, etc. A beer can be manufactured by passing through the process which removes, a fermentation process, an aging process, etc. When the beer-taste beverage is a non-alcoholic beer, for example, after the step of obtaining wort from malt, the step of adding various additives, fragrances, etc. to the obtained wort, the non-alcoholic beer Can be manufactured.

  In the said improvement method, you may use barley raw materials other than LOX-1 deletion barley raw material, ie, the barley raw material which has not deleted LOX-1. As the barley materials other than the LOX-1 deficient barley materials, any materials suitable for producing beer-taste beverages can be used.

  In the method for improving the astringency of beer-taste beverages, the ratio of LOX-1-deleted barley materials in all barley materials used for producing beer-taste beverages may be 10% or more, and 20% or more. Preferably, it is 30% or more, more preferably 50% or more, still more preferably 75% or more, and particularly preferably 85 to 95%. The ratio of the LOX-1 deficient barley raw material in the total barley raw material used is, for example, 10 to 100%, 20 to 100%, 20 to 95%, 30 to 95%, 50 to 95%, 75 to 95%. It may be.

  In the method for improving mildness of beer-taste beverages, the ratio of LOX-1 deficient barley materials in all barley materials used for producing beer-taste beverages may be 10% or more, and 20% or more. Preferably, it is 30% or more, more preferably 50% or more, still more preferably 75% or more, and particularly preferably 85 to 95%. The proportion of the LOX-1 deficient barley material in the total barley material used may be, for example, 10-100%, 20-95%, 30-95%, 50-95%, 75-95%. The above ratio is based on mass.

  You may use fermentation raw materials other than barley raw material for manufacture of a beer taste drink. Examples of fermentation raw materials other than barley raw materials include starch raw materials such as rice, corn, starch, corn grits, wheat and corn, and sugar raw materials such as liquid sugar and sugar. In addition, the beer-taste beverage may contain an antioxidant, a colorant, carbon dioxide gas and the like as other auxiliary materials.

  The beer-taste beverage may be provided in a container. As a container, the well-known thing used for alcoholic beverages can be used, For example, plastic containers, such as a can, a bottle, a PET bottle, a paper container, a pouch container, etc. are mentioned.

  Hereinafter, based on an Example, this invention is demonstrated more concretely. However, the present invention is not limited to the following examples.

[Test Example 1]
(Barley)
LOX-1 deficient barley and barley having LOX-1 activity are malted by slaughtering, germination, and roasting using a wheat making apparatus made by Phoenix, LOX-1 deficient malt and LOX- A malt having 1 activity was obtained. The LOX-1-deficient barley used in the test examples of the present specification is barley LOX-1 in which the guanine at the splicing donor site (5′-GT-3 ′) of the fifth intron of barley LOX-1 gene is mutated to adenine. Barley with a mutated gene.

(Preparation)
LOX-1 deficient malt was mixed with malt having LOX-1 activity in the proportions shown in Tables 1 and 2 below, so that the total amount of malt was 1080 g. The malt was pulverized, mixed with 4 L of water for preparation to which 0.924 g of calcium sulfate was added, saccharified, and the produced saccharified solution was filtered to obtain wort. To 3.5 L of the obtained wort, 4.6 g of hops and 383 μL of lactic acid were added and boiled, and the wort was cooled and filtered.

(fermentation)
Yeast was added to the obtained filtrate for fermentation to obtain beer.

  About the astringency and mildness of the obtained beer, sensory evaluation was performed by the expert panel. The evaluation in Sample 1 in which the ratio of LOX-1 deficient malt was 0% of the total malt was evaluated as a standard (3.0), and was evaluated with a maximum of 5 points. The results are shown in Tables 1 and 2. Similarly, the overall evaluation of beer was evaluated using Sample 1 as a reference (3.0). The evaluation of astringency indicates that the higher the number, the less astringency. The higher the numerical value, the milder the mildness. The higher the numerical value, the better the beverage.

  Sample 1 using only LOX-1 deficient malt as malt had astringency and bitterness and was inferior to mild. On the other hand, samples 2-8 produced using at least a portion of the LOX-1-deleted malt improved mildness, while samples 3-8 had reduced astringency. In particular, Sample 7 in which the proportion of malt lacking LOX-1 in the malt was 90% was excellent in both astringency and mildness and well-balanced.

[Test Example 2]
The total amount of malt is 868 g, and 346.2 g of rice, starch and corn grits are combined as fermentation raw materials other than barley raw materials, and the ratio of the LOX-1 deficient malt amount in the total amount of malt is the value shown in Table 3 Prepared beer in the same manner as in Test Example 1.

  About the astringency and mildness of the obtained beer, sensory evaluation was performed in the same manner as in Test Example 1 by using an expert panel with reference to Sample 9. The results are shown in Table 3. Even when fermenting raw materials other than barley were blended, a tendency that mildness was improved and astringency was reduced when the proportion of the LOX-1-deficient barley raw material in the barley raw material was large to some extent was confirmed.

[Test Example 3]
As barley raw materials, 23.1% (254g) of malt and 76.9% (847g) of barley are mixed, the ratio of LOX-1 deletion malt amount in the total amount of malt, and LOX-1 deletion in the total amount of barley A beer-taste beverage was obtained by preparing in the same manner as in Test Example 1 except that the barley content was set to the values shown in Table 4.

  About the astringency and mildness of the obtained beer taste drink, sensory evaluation was performed like the test example 1 by the expert panel on the basis of the sample 15. The results are shown in Table 4. Even when malt and barley were used in combination as barley raw materials, an improvement in mildness and a reduction in astringency were observed when the proportion of the LOX-1-deficient barley raw material in the whole barley raw material was large to some extent. Sample 19 was excellent in both astringency and mildness, and had the best balance.

Claims (8)

  A beer-taste beverage astringency improving agent comprising a lipoxygenase-1-deficient barley material as an active ingredient.   A mildness improving agent for beer-taste beverages comprising a lipoxygenase-1 deficient barley material as an active ingredient.   A method for improving the astringency of a beer-taste beverage, comprising using a lipoxygenase-1-deficient barley material as a barley material.   The astringency improvement method of Claim 3 whose ratio of the said lipoxygenase-1 deficient barley raw material in a whole barley raw material is 20% or more.   The method for improving astringency according to claim 3 or 4, wherein the barley raw material is malt and includes a step of obtaining wort from the malt.   A method for improving the mildness of a beer-taste beverage, comprising using a lipoxygenase-1 deficient barley material as a barley material.   The mildness improvement method of Claim 6 whose ratio of the said lipoxygenase-1 deficient barley raw material in a whole barley raw material is 10% or more.   The method for improving mildness according to claim 6 or 7, wherein the barley raw material is malt and includes a step of obtaining wort from the malt.