JP2010148485A - Fermented alcoholic beverage with high content of ferulic acid - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for producing a fermented alcoholic beverage high in functionality and rich in flavor and taste, in which, to give the beverage functionality, the content of free ferulic acid is increased and yet a balance of fermented alcoholic beverages between flavor and taste is kept in fermented alcoholic beverages such as beer, sparkling wine or other brewage. <P>SOLUTION: In the production of fermented alcoholic beverages using at least one selected from barley, wheat, soybean, barley malt and wheat malt as fermentation material, the content of free ferulic acid is increased in a saccharification process under the presence of an enzyme having a ferulic acid esterase activity, an enzyme having a cellulase activity and an enzyme having a xylanase activity by using the saccharification temperature pattern in which the saccharification temperature is 45°C±10°C and the time for protein degrading is not less than 5 min; the formation of 4-VG(4-vinyl guaiacol) is repressed by fermenting with yeast having a low ability of generating 4-VG; and fermented alcoholic beverage keeping a balance between flavor and taste is produced. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention is a beer, sparkling liquor, or other brewed liquor that contains a high content of free ferulic acid and suppresses the production of 4-vinyl guaiacol (4-VG) and maintains the flavor balance of the fermented alcoholic beverage. It is related with the manufacturing method of such fermented alcoholic beverages. Furthermore, the present invention relates to a method for producing a baitzen beer with reduced baitzen fragrance and high free ferulic acid content.

  Due to the recent health-oriented boom, there has been an increasing interest in the functionality of foods, and the development of health-conscious products is also expected in fermented alcoholic beverages such as beers. With regard to the health functionality of beer, research has been conducted to scientifically approach the functionality of beer and its raw materials from the viewpoint of preventing lifestyle-related diseases (brewing association magazine, 98, 228-240, 2003). Antioxidant properties of beer have been studied. When compared in vitro, beer is about one-fifth of alcoholic equivalent red wine, but is given to rat blood after oral administration. It has been shown that the antioxidant activity was about the same. This suggests that some antioxidant component in beer may exhibit a high in vivo antioxidant effect, but it is not clear what this substance is.

  On the other hand, ferulic acid is known as a component having a health function. Ferulic acid is a phenolic compound contained in a large amount in wheat bran, rice bran and the like. In plant tissues, ferulic acid is present in the cell wall, and has a function of regulating the growth of plants and suppressing the invasion of pathogenic microorganisms by forming a bridge between cell wall polysaccharides. In the plant cell wall, ferulic acid is present as an ester bond with sugars such as arabinose, but ferulic acid esterase can be hydrolyzed by ferulic acid esterase to release ferulic acid. In alcoholic beverages, this free ferulic acid is decarboxylated by ferulic acid decarboxylase, which is a microbial enzyme, to produce a unique aroma called 4-zen guaiacol (4-VG) -derived baizen fragrance.

  The functionality of ferulic acid is said to have an antioxidant effect by donating hydrogen to free radicals through phenolic —OH groups. Because of these properties, it is used as an antioxidant and has been reported to have functions such as lipid metabolism improvement, anti-inflammatory, dementia, diabetic nephropathy prevention, and whitening action. It has been reported that phenolic acid including ferulic acid is actively and efficiently absorbed by a transport system like a monocarboxylic acid transporter (MCT), and ferulic acid with excellent in-vivo and absorbability is reported. It is getting attention (Chemistry and Biology, 44, 532-538, 2006). Although in vitro antioxidant activity of ferulic acid is not high, it is expected to exhibit a higher antioxidant function in vivo due to its excellent in vivo utilization and absorbability. However, there is actually no report comparing the antioxidant activity of beer with a high ferulic acid content.

  Numerous studies aiming at high ferulic acid content have been conducted. For example, research has been conducted on the production of beer characterized in that the release of ferulic acid from malt is promoted by adding cell wall degrading enzymes in the saccharification process and the content of phenolic compounds is high (Japan). Abstracts of Agricultural Chemistry Conference, 2004). However, although research on enzymes relating to ferulic acid production has been conducted, preparation conditions and flavor balance have not been studied, and the relationship between ferulic acid high content and antioxidant activity has not been studied. In addition to alcoholic beverages, in addition to knowledge about cereal ferulic acid, investigations have been reported on the relationship between ferulic acid fate and antioxidant activity in the malting process of wheat miso (Brewing Association Journal, 96, 100-106, 2001).

  On the other hand, conventionally, distilled liquor has a technology for producing a distilled liquor with a rich taste by adding ferulic acid or producing free ferulic acid in the production process as an improvement in taste, and therefore A process for producing free ferulic acid is disclosed. That is, in distilled liquor, as a mechanism of flavor generation, ferulic acid esterified with arabinoxylan, which constitutes the cell wall of raw cereals, is liberated in the mash by esterases such as koji molds, and this ferulic acid is released during atmospheric distillation. It is decarboxylated by heating to 4-VG, and it is distilled in the resulting distilled liquor. And 4-VG contained in distilled liquor has the mechanism that it is oxidized during the storage and turns into vanillin, and gives a rich taste to distilled liquor. Therefore, various methods have been disclosed as techniques for promoting the mechanism of taste generation.

  For example, Japanese Patent Application Laid-Open No. 6-62829 discloses a method for improving mild distilled liquor by adding acetic acid, succinic acid, syringic acid, ferulic acid, isoamyl alcohol and the like to distilled liquor. Further, JP-A-9-238673 discloses that alcohol concentration is performed using Saccharomyces cerevisiae TSH-1 having ferulic acid decarboxylase to increase the 4-VG concentration in the mash and distill after storage. JP-A-10-276788 discloses a method for producing a distilled liquor such as shochu (Awamori) having a rich flavor by increasing vanillin in the liquor. JP-A-10-276788 discloses a ferulic acid decarboxylase gene. A method for producing a distilled liquor with excellent flavor using the introduced yeast is disclosed.

  Furthermore, JP-A-2000-125840 discloses a method for producing a shochu with a sweet and rich flavor and a rich body by using a yeast having a high ferulic acid decarboxylation activity as a yeast in the production of shochu. JP-A-2008-75 discloses a method for producing a distilled liquor such as awamori with a rich flavor by acting on Lactococcus lactis having an enzyme that converts ferulic acid into 4-VG. No. 2004-236634 discloses a koji mold having a xylanase activity of 13.5 U / g 麹 or more and a ferulic acid esterase activity of 10.0 U / g 麹 or more as koji molds used for koji making in a method for producing cereal distilled liquor. A method for producing a high-flavored cereal distilled liquor by using the above has been developed.

  In order to improve the taste in these distilled spirits, either ferruleic acid is added in the production process or free ferulic acid is produced to produce a rich taste distilled spirit. It is expected that ferulic acid is converted into 4-VG or vanillin by distillation or the like in the manufacturing process or in the storage process. However, such a method is, for example, beer or sparkling liquor. It is difficult to apply to the production of fermented alcoholic beverages where production and product management are performed under anaerobic conditions.

  In addition, it has been disclosed to improve the flavor of sake such as sake by using ferulic acid or the like. For example, in JP-A-8-89230, in a mash saccharification and fermentation ripening process of sake, hydroxycinnamate ester hydrolase is added to act to add ferulic acid, ferulic acid ethyl ester, vanillin, vanillic acid, etc. A method for producing sake with a rich flavor component is disclosed. Further, JP-A-2007-202504 discloses the use of a plant cell wall degrading enzyme agent having a ratio of xylanase activity / ferulic acid esterase activity of 4.1 or more when producing sake such as sake. It has been disclosed to produce alcoholic beverages excellent in liquor quality containing a high content of ferulic acid even when treated with activated carbon. In this disclosed method, when raw materials having a low koji and fine polishing ratio are used to make fermented acid contain a high amount of ferulic acid, ferulic acid is adsorbed at the same time when adsorbing and removing lipids and proteins brought in by activated carbon treatment. Although it is prevented from being removed and ferric acid is highly contained in alcoholic beverages, such ferulic acid is a sugar-binding type and is not free type ferulic acid.

  In these alcoholic beverages such as sake, the method used to make ferulic acid and the like highly contained is also a method targeting alcoholic beverages such as sake, as described above, such as beer and sparkling sake. It is not directly applicable to the production of a fermented alcoholic beverage.

JP-A-6-62829. JP-A-8-89230. JP-A-9-238673. Japanese Patent Laid-Open No. 10-276788. JP 2000-125840 A. JP-A-2004-236634. Japanese Patent Application Laid-Open No. 2007-202504. Japanese Patent Application Laid-Open No. 2008-75. Journal of the Brewing Society, 98, 228-240, 2003. Chemistry and Biology, 44, 532-538, 2006. Abstracts of the Japanese Agricultural Chemistry Conference, 2004. Journal of the Brewing Society, 96, 100-106, 2001.

  An object of the present invention is to provide a fermented alcoholic beverage such as beer, happoshu, or other brewed liquor with a high content of free ferulic acid in order to impart functionality to the beverage, while maintaining the flavor balance of the fermented alcoholic beverage An object of the present invention is to provide a method for producing a fermented alcoholic beverage with high functionality and high flavor.

  That is, conventionally, in spirits and the like, ferulic acid is released and released by acting esterase etc. on ferulic acid ester-bonded with arabinoxylan which constitutes the cell wall of the raw cereal for improving taste and the like. A method for increasing the fatty acid content is disclosed. However, when the method is used as a method for adding high ferulic acid to fermented alcoholic beverages such as beer, happoshu, or other brewed liquor and imparting functionality to the beverage, free ferula Although the functionality can be imparted to the fermented alcoholic beverage by containing the acid in a high amount, there is a problem that the flavor balance of the fermented alcoholic beverage cannot be maintained and a high flavored fermented alcoholic beverage cannot be produced. Then, in the earnest examination about the cause, in producing fermented alcoholic beverages, 4-VG (4-vinyl produced in the reaction of enzyme reaction that releases ferulic acid ester-bonded with arabinoxylan or fermentation process. We found out that there was a cause in (Guaiacol).

  That is, as already explained, as a taste improvement in distilled liquor, all the methods for producing free ferulic acid in the production process to produce a rich taste distilled liquor are in the production process of the distilled liquor. It relies on the conversion of ferulic acid from 4-VG to vanillin, for example by oxidation or the like, during distillation or storage, and thus such a process is anaerobic, such as beer or happoshu. Even if it is applied to the production of fermented alcoholic beverages where production and product management are performed under the conditions, the conversion from 4-VG to vanillin is not performed, and the ferulic acid content is high and the flavor balance is maintained. A flavored fermented alcoholic beverage cannot be produced. 4-VG is called a baizen fragrance, and is a flavor component of baizen beer produced using wheat malt. However, when this component increases, the flavor balance of the fermented alcoholic beverage cannot be maintained.

  Therefore, the subject of the present invention is a fermented alcoholic beverage such as beer, happoshu, or other brewed liquor, and in order to impart functionality to the beverage, the content of free ferulic acid is high, and the flavor of the fermented alcoholic beverage A method for producing a fermented alcoholic beverage that effectively increases the amount of free ferulic acid in the fermented alcoholic beverage and suppresses the production of 4-VG in order to produce a highly functional and highly flavored fermented alcoholic beverage that maintains a balance. It is to provide. Here, “other brewed liquor” refers to “other miscellaneous liquor” that has been changed to “other brewed liquor” due to the revision of the Liquor Tax Law. “Miscellaneous sake” possessed is classified into “Happoshu” using wheat or malt as a raw material and “Other brewed liquors” other than Happoshu. “Other brewed liquors” do not use wheat or malt, but break down plant proteins such as beans and cereals with enzymes to obtain the necessary nitrogen source and ferment it by adding saccharified liquid. is there. Therefore, except for this point, the method for producing “other brewed liquor” is not different from the method for producing beer or happoshu.

  In order to solve the above-mentioned problems, the present inventor effectively increases free ferulic acid in a fermented alcoholic beverage, suppresses the production of 4-VG, and maintains a balanced flavor. In the intensive study of beverage production methods, barley, wheat, soybeans, barley malt, and wheat malt are used as brewing raw materials to produce fermented alcoholic beverages. , Select the amount of added enzyme, set the optimal conditions for ferulic acid to be released from arabinoxylan in the raw material, and ferment using yeast with low 4-VG production ability, the free ferulic acid in fermented alcoholic beverages It is possible to increase the production of highly functional and highly flavored fermented alcoholic beverages that are effectively increased and that suppress the production of 4-VG and maintain a flavor balance. It found, which resulted in the completion of the present invention.

  In the present invention, the saccharification temperature pattern for effectively increasing the free ferulic acid in the fermented alcoholic beverage is such that ferulic acid is produced from arabinoxylan in the malt as a temperature condition for producing a large amount of ferulic acid as a precursor of 4-VG. A saccharification pattern was set in which the temperature was lowered from 20 to 60 ° C. and the protein resting time was 5 minutes or longer in consideration of the optimum temperature of 45 ° C. to be released. That is, the present invention provides a saccharification step in the production of a fermented alcoholic beverage using any one or more of barley, wheat, soybean, barley malt, and wheat malt as a brewing material, an enzyme having ferulic acid esterase activity, and cellulase activity. In the presence of an enzyme having a glycanase activity and an enzyme having a xylanase activity, using a saccharification temperature pattern with a saccharification temperature of 45 ° C. ± 10 ° C. and a protein resting time of 5 minutes or more, and fermenting using yeast with low 4-VG production By this, it consists of the method of manufacturing the fermented alcoholic beverage which contains high free ferulic acid and the flavor balance was hold | maintained.

  In the present invention, examples of preferable enzymes include protease MG as an enzyme having ferulic acid esterase activity, laminex super as an enzyme having cellulase activity, and cellulosin BGL as an enzyme having xylanase activity. In the present invention, the content of free ferulic acid can be further increased by adding wheat bran to the brewing raw material. Furthermore, in the present invention, the amount of free ferulic acid in wort can be increased by increasing the amount of enzyme added, and the amount of free ferulic acid in wort can be increased to 30 mg / L. it can.

  In the present invention, maintaining the flavor balance of a fermented alcoholic beverage with an increased content of free ferulic acid is achieved by inhibiting the production of 4-VG by fermenting with yeast having a low 4-VG production ability. The amount of 4-VG produced can be suppressed to 0.62 mg / L or less by applying the method of the present invention. Examples of the fermented alcoholic beverage produced by the production method of the present invention include beer, happoshu, and other brewed liquors. By applying the method for producing a fermented alcoholic beverage of the present invention to the production of a baitzen beer, a baitzen beer can be produced which has a reduced baitzen fragrance and contains a high amount of free ferulic acid.

  That is, the present invention specifically relates to (1) the production of a fermented alcoholic beverage using any one or more of barley, wheat, soybeans, barley malt and wheat malt as a brewing raw material. In the presence of an enzyme having an enzyme, an enzyme having a cellulase activity, and an enzyme having a xylanase activity, using a saccharification temperature pattern of 45 ° C. ± 10 ° C. and a protein resting time of 5 minutes or more. A method for producing a fermented alcoholic beverage containing a high acid content and maintaining a flavor balance; and (2) an enzyme having ferulic acid esterase activity is protease MG, an enzyme having cellulase activity is laminex super, and xylanase The release according to (1) above, wherein the enzyme having activity is cellulosin BGL A method for producing a fermented alcoholic beverage containing a high amount of erula acid and maintaining a flavor balance, and (3) the release according to (1) or (2) above, wherein wheat bran is added to the brewing material A method for producing a fermented alcoholic beverage containing a high content of ferulic acid and maintaining a flavor balance, and (4) the amount of free ferulic acid is 30 mg / L or more, (1) to (3) A method for producing a fermented alcoholic beverage containing a high content of free ferulic acid and maintaining a flavor balance.

  Moreover, this invention increases the addition amount of (5) enzyme until the amount of free ferulic acids in wort becomes 30 mg / L, The release in any one of said (1)-(4) characterized by the above-mentioned. A method for producing a fermented alcoholic beverage containing a high amount of ferulic acid and maintaining a flavor balance, and (6) maintaining the flavor balance comprises suppressing the production of 4-vinyl guaiacol in the fermentation process. A method for producing a fermented alcoholic beverage that contains a high amount of the free ferulic acid according to any one of the above (1) to (5) and that maintains a flavor balance, and (7) the amount of 4-vinylguaiacol produced is 0. The method for producing a fermented alcoholic beverage having a high content of free ferulic acid as described in the above (6) and maintaining a flavor balance, characterized by being suppressed to .62 mg / L or less, and (8) fermentation The alcohol beverage is beer, happoshu, or other brewed liquor, which contains a high content of free ferulic acid according to any one of (1) to (7) above, and maintains a flavor balance A method for producing a fermented alcoholic beverage, (9) a high content of free ferulic acid as described in (8) above, characterized in that it is a baitzen beer in which the baizen aroma is suppressed and the content of free ferulic acid is high. , A method for producing a fermented alcoholic beverage that maintains a flavor balance.

  According to the present invention, in fermented alcoholic beverages such as beer, happoshu, or other brewed liquor, high-functional, high-flavored fermented alcoholic beverages that contain a high content of free ferulic acid and that maintain the flavor balance of fermented alcoholic beverages, And a manufacturing method thereof. By applying the method for producing a fermented alcoholic beverage according to the present invention to a method for producing a baizen beer, to provide a baitzen beer with a high function and a high flavor that has a high taste and a high flavor and that contains a high amount of free ferulic acid with a low baizen aroma. Can do.

  The present invention relates to a saccharification step in the production of a fermented alcoholic beverage such as beer, happoshu, or other brewed liquor using any one or more of barley, wheat, soybean, barley malt, and wheat malt as a brewing raw material. In the presence of an enzyme having ferulic acid esterase activity, an enzyme having cellulase activity, and an enzyme having xylanase activity, using a saccharification temperature pattern of 45 ° C. ± 10 ° C. and a protein resting time of 5 minutes or more. Fermentation alcohol with high functionality and high flavor by maintaining the flavor balance by suppressing the production of 4-VG by fermenting with yeast having a high content of free ferulic acid and low 4-VG production ability Consisting of producing a beverage.

  In the saccharification step in the method for producing a fermented alcoholic beverage according to the present invention, ferulic acid is liberated from arabinoxylan in the brewing raw material, and the enzyme used for producing high ferulic acid content in wort is ferulic acid esterase activity. A combination of an enzyme having an enzyme, an enzyme having cellulase activity, and an enzyme having xylanase activity is used. Moreover, in the enzyme reaction in a fermentation process, use of yeast with low 4-VG production ability is desirable for the purpose of suppressing production of 4-VG. Examples of the yeast include bottom fermentation yeast KBY011.

  Protease MG (protease, peptidase, ferulic acid esterase) (protease M Amano G: Amano Enzyme Co., Ltd.); Laminex Super (cellulase (β-glucanase) (Danisco Japan Co., Ltd.)); Co.) is a commercially available enzyme and can be obtained under the trade name of the enzyme used in the method for producing a fermented alcoholic beverage of the present invention. Examples of the added amount of enzyme include kg malt, laminex super 0.5 g / kg malt, and cellulosin BGLg / kg malt.

  In the method for producing a fermented alcoholic beverage of the present invention, barley, wheat, soybeans, barley malt, and wheat malt are used as brewing raw materials for producing free ferulic acid. Further, bran, brown rice, and polished rice are used. Can be added. In cereals and food materials made from cereals, ferulic acid is contained in large amounts in the order of brown rice, germ-milled rice, and polished rice in rice, and in wheat and in germ and brown rice. Since ferulic acid is mainly contained in the cell wall, it is considered that the ferulic acid increases as the cell wall content increases. Moreover, in malt, the production amount of ferulic acid increases when barley malt is used as a raw material rather than wheat malt. Therefore, when producing a fermented alcoholic beverage using malt, the amount of ferulic acid produced can be adjusted by appropriately changing the blending ratio of barley malt and wheat malt as a brewing raw material.

  In the method for producing a fermented alcoholic beverage of the present invention, the amount of enzyme added to produce free ferulic acid can be appropriately determined. In the present invention, as the amount of enzyme added increases, the amount of free ferulic acid produced increases in a linear relationship up to about 30 mg / L. When it exceeded about 30 mg / L, a moderate increase in free ferulic acid concentration was observed with an increase in the amount of enzyme. In the present invention, the amount of free ferulic acid can be produced up to 34 mg / L by increasing the amount of enzyme.

  In the method for producing a fermented alcoholic beverage of the present invention, wheat bran can be added to the brewing raw material in order to increase the production of free ferulic acid. Since wheat bran contains a lot of ferulic acid, adding fermented wheat bran in addition to brewing materials such as barley malt and wheat malt increases the content of free ferulic acid in wort Can do. By applying the method for producing a fermented alcoholic beverage according to the present invention, wheat malt and barley malt are mixed and used to optimize the saccharification process and ferment using a yeast having a low 4-VG production ability, thereby producing a baizen fragrance. It is possible to produce a high-function, high-flavored new taste of Bautzen beer with a high content of free ferulic acid.

  In the method for producing a fermented alcoholic beverage according to the present invention, as a saccharification step condition for effectively increasing the free ferulic acid in the fermented alcoholic beverage, the saccharification temperature pattern is produced in a large amount of ferulic acid that is a precursor of 4-VG. As a temperature condition, an optimum condition for releasing ferulic acid from arabinoxylan in the malt, a saccharification temperature of 20 to 60 ° C. and a protein resting time of 5 minutes or more in consideration of a saccharification temperature of 45 ° C. were set. That is, a saccharification pattern having a saccharification temperature of 45 ° C. ± 10 ° C. and a protein rest time of 5 minutes or more is used as the saccharification temperature pattern. The protein rest time is 5 minutes or longer, usually 5 to 60 minutes, and 5 to 120 minutes at the maximum.

  In the method for producing a fermented alcoholic beverage according to the present invention, the conditions for the saccharification step for effectively increasing free ferulic acid in the fermented alcoholic beverage are the conditions optimized for the brewing raw material, the type of enzyme to be added, the amount of the added enzyme, and the saccharification temperature. In addition, there is no difference from the processing method and processing conditions of the saccharification and fermentation processes in the production of ordinary fermented alcoholic beverages, except that the use of yeast having a low 4-VG production capacity in the fermentation process is adopted. The processing method and processing conditions of the fermented alcoholic beverage manufacturing process other than the saccharification and fermentation processes are the same as the normal fermented alcoholic beverage manufacturing process.

  EXAMPLES Hereinafter, although an Example demonstrates this invention more concretely, the technical scope of this invention is not limited to these illustrations.

  <Test method>

[500 ml saccharification test]
As a 500 ml saccharification test, a saccharification test was conducted using the following saccharification temperature pattern: a total amount of 80 ml of barley malt or wheat malt was added with 320 ml of hot water and protease MG 0.2 g / kg malt, lami Enzyme was added so that nexus super 0.5 g / kg malt and cellulosin BGL 0.5 g / kg malt were added, and saccharification treatment was performed. As the saccharification temperature pattern, a temperature condition in which a large amount of ferulic acid as a precursor of 4-VG was produced was used. A saccharification pattern was used in consideration of a saccharification temperature of 45 ° C. ± 10 ° C. at which ferulic acid is released from arabinoxylan in the malt and a protein rest time of 5 minutes or more.

[200L saccharification test]
As a 200 L saccharification test, a saccharification test was performed using the following saccharification temperature pattern: protease malt 0.2 mg / kg malt, laminex super 0.5 g / kg malt for a total of 50 kg of barley malt or wheat malt. Enzyme treatment was performed by adding enzyme so that the cellulosin BGL was 0.5 g / kg malt. Similar to the small-scale saccharification test, a saccharification pattern in which ferulic acid was released from arabinoxylan in the malt at a saccharification temperature of 45 ° C. ± 10 ° C. and a protein rest time of 5 minutes or more was used.

[Fermentation test]
Fermentation tests were performed using worts prepared in the 500 ml saccharification test and the 200 L saccharification test: In the 500 ml fermentation test, 2.5 g of bottom fermentation yeast KBY011 strain was added to 500 ml of wort, and after aeration, the temperature was increased. A fermentation test was conducted at 20 ° C. for 7 days. In the 200 L fermentation test, 0.52% yeast was added to 200 L of wort.

[Measurement of ferulic acid concentration]
In this example, the concentration of free ferulic acid and sugar-bound ferulic acid were measured by the following method.

[Method for measuring concentration of free ferulic acid]
To measure the concentration of free ferulic acid, 5 ml of water and 50 ml of a 1 mol / L aqueous sodium hydroxide solution were added to 1 g of the sample used, followed by hydrolysis at 40 ° C. for 3 hours. About 28 ml of 2 mol / L hydrochloric acid and 150 ml of ethyl acetate were added and shaken for 15 minutes. After separating the ethyl acetate layer, it was dehydrated and dried, and 5 ml of a mixed solution of methanol and water (1: 1) was added, followed by membrane filter filtration. Then, it used for the high performance liquid chromatograph. The operating conditions of the high performance liquid chromatograph are as follows.
<Operating conditions of high performance liquid chromatograph>
Column: YMC-Pack ODS-A AA12S05-1506WT, φ6.0 × 150mm
Mobile phase: Acetic acid and acetonitrile mixed liquid flow rate: 1.0 ml / min
Column temperature: 40 ° C
Measurement wavelength: 320 nm

[Measurement of 4-VG concentration]
The measurement of 4-VG concentration was analyzed by GC (gas chromatography) after pretreatment.

[Antioxidant activity measurement]
For the measurement of antioxidant activity, ORAC (Oxygen Radical Absorbance Capacity) method (J. Agric. Food Chem., 53,4290-4302, 2005) was used. The ORAC method is a representative method for measuring the antioxidant power of foods, and is said to be able to measure the antioxidant power against peroxyl radicals, which are the main free radicals in the living body. The higher the ORAC value, the higher the scavenging ability (antioxidant power) of active oxygen. In serum ORAC measurement, pretreatment with perchloric acid was performed to eliminate the influence of blood proteins (J. Agric. Food Chem., 54, 7940-7946, 2006). Serum ferulic acid concentration was measured according to the method of Konishi et al. (J. Agric. Food Chem., 53, 9928-9933, 2005).

  <Test and results>

  In considering the development of ferulic acid-rich beers, we first analyzed how much ferulic acid was contained in existing beers. The analyzed beers were German Bizen Beer [Products A, B, C], Japanese wheat malt beer [Products D, E, F], and as a control normal beer [Product G] and Happoshu [ There are 8 types of products H]. The analysis items are free ferulic acid and 4-VG.

  As a result, as shown in FIG. 1 (concentration of free ferulic acid and 4-VG in product beer), German Bizen beer products A, B and C contain a large amount of 4-VG, but free ferulic acid. Was hardly included. Japanese wheat malt beer products D, E, F Normal beer product G and Happoshu product H contain about 2 mg / L of free ferulic acid, but have a 4-VG concentration compared to German baitzen beer. Was low. From the above results, it was suggested that the concentrations of free ferulic acid and 4-VG were greatly different between the investigated Bezen beer and the other beers.

[Enzyme content and free ferulic acid production]
The effect of enzyme amount on the formation of free ferulic acid was tested. In ferulic acid, ferulic acid bound to arabinoxylan is decomposed into free ferulic acid by ferulic acid esterase. Using a mixture of the proteolytic enzyme protease MG containing the ferulic acid esterase and the hemicellulase enzymes laminex super and cellulosin BGL, the effect of the enzyme amount on ferulic acid production was tested. Added to 80 g of barley malt, protease MG 0.2 g / kg malt, laminex super 0.5 g / kg malt, cellulosin BGL 0.5 g / kg malt, saccharification temperature 45 ° C. ± 10 ° C., protein rest time Ferulic acid was liberated using saccharification conditions for 5 minutes or longer.

  As a result, as shown in FIG. 2, the ferulic acid concentration increased in a linear relationship with the amount of free ferulic acid up to about 30 mg / L as the amount of enzyme increased. When it exceeded about 30 mg / L, a moderate increase in free ferulic acid concentration was observed with an increase in the amount of enzyme. From this result, it was suggested that it is possible to produce free ferulic acid concentration up to about 34 mg / L by further increasing the amount of enzyme.

[Effect of enzyme type on free ferulic acid production]
The effect of enzyme type on the formation of free ferulic acid was tested. Ferulic acid esterase that liberates ferulic acid is not a commercially available enzyme by itself. Contained in hemicellulase and proteolytic enzymes. Therefore, the effect of enzyme type on ferulic acid production was examined. Ferulic acid-producing ability was evaluated for 8 types of enzymes (Amano 3G, Neutase, HP, Protease MG, Sumiteam LPL, Sumiteam FP-G, Umamizyme G, Bromelain F).

  As a result, when protease MG was used as shown in FIG. 3 (enzyme type and production of free ferulic acid), ferulic acid production was increased as compared to the control without enzyme. Protease MG is a proteolytic enzyme, but also possesses ferulic acid esterase. Further, in FIG. 4 (protease MG, laminex super, cellulosin BGL enzyme alone and in combination and production of free ferulic acid), the enzyme alone affects the production of free ferulic acid for three types of protease MG, laminex super, and cellulosin BGL. The contribution was tested. As a result, it has been clarified that Amano MG and Laminex super enzymes have high activities and contribute greatly to the production of free ferulic acid. It has been shown that high free ferulic acid production can be achieved by mixing these three enzymes.

[Wheat malt use ratio and free ferulic acid production]
Wheat malt use ratio and free ferulic acid production were tested. The ferulic acid content in cereals and food materials and processed foods made from cereals was investigated, and in rice, brown rice, germ-milled rice, and milled rice were contained in this order, and in wheat, germ and brown grain were found to contain more (Brewing association magazine, 96, 100-106, 2001). Since ferulic acid is mainly contained in the cell wall, it is estimated that the ferulic acid increases as the cell wall content increases. Therefore, the effect of wheat malt use ratio on ferulic acid production was investigated. As a result, contrary to prediction, ferulic acid production decreased as the wheat malt use ratio increased (FIG. 5). That is, it was shown that the amount of ferulic acid produced increases under this condition when barley malt is used as a raw material rather than wheat malt.

[Antioxidant activity of saccharified liquid containing high free ferulic acid]
In vitro antioxidant activity of beer with high free ferulic acid content was measured by ORAC method. As shown in FIG. 6 (in vitro antioxidant activity measurement results of beer with a high ferulic acid content), beer with a high ferulic acid content (beer with high ferulic acid content) by the production method described above is obtained by using the same raw materials. It was confirmed that the antioxidant power was enhanced by 50% or more as compared with beer produced by the general production method used (control beer). Then, the ORAC value in the blood when the ferulic acid-rich beer and the control beer were orally forcibly administered was measured over time. Each beer was freeze-dried, redissolved in water, and administered to rats (Wistar strain, male 8 weeks old) to a concentration of 0.58 g / kg body weight (3 mL). As shown in FIG. 7 (in vivo antioxidant activity measurement result of ferulic acid-rich beer), compared to the control beer-administered group, the ORAC value was significantly increased in the ferulic acid-rich beer-administered group. It was also shown that it is excellent in enhancing the antioxidant power in vivo. In addition, as shown in FIG. 8 (blood ferulic acid concentration in rats orally administered with beer containing high ferulic acid), the ferulic acid concentration in the rat serum 20 minutes after the ORAC value markedly increased was measured. A higher ferulic acid concentration was detected in the ferulic acid-rich beer group compared with the control beer group.

[Production of Bizen Beer High in Free Ferulic Acid Containing Bitzen Aroma]
A bezen beer with high free ferulic acid content with reduced baizen fragrance was produced.
Bautzen beer is a top-fermented beer that is often drunk mainly in the state of Bavaria in Germany. Its characteristic is that it uses more than 50% of wheat malt as a raw material, and it is unique called ester fragrance and 4-VG derived baizen fragrance. It is a point with the scent of.

  Therefore, for the purpose of producing a baitzen beer with a reduced baizen fragrance and high ferulic acid content, 70% wheat malt is used, and 4-VG precursor (free ferulic acid) is produced in a large amount. In order to suppress the production, 200L saccharification test and fermentation test were conducted under fermentation conditions using the bottom fermenting yeast KBY011 strain to produce a test brewed beer. As a result, the brewed beer contained 9 mg / L of free ferulic acid (4.5 times the commercial product), and the 4-VG content was 0.62 mg / L (about 1/4 of German Bizen beer). (FIG. 9: concentration of free ferulic acid and 4-VG in baitzen beer with high free ferulic acid content with suppressed baitzen fragrance). That is, by this method, a bezen containing a high concentration of ferulic acid was able to be produced while suppressing the baizen fragrance. As a result of tasting the prepared beer, although it felt slightly astringent, it was possible to produce a new flavored Bizen beer with a low phenolic aroma, a reduced baizen aroma, and a high ferulic acid content.

[Increase of free ferulic acid content of beer by adding wheat bran]
The increase in free ferulic acid content of beer by adding wheat bran was tested. Wheat bran is thought to contain a lot of ferulic acid. Thus, the effects of ferulic acid content of wheat, wheat malt, wheat bran and barley malt and the addition of wheat bran on ferulic acid production were examined.

  As a result, as shown in FIG. 10 (average free ferulic acid concentration (mg / L) per 11 worts of ferulic acid content of wheat, wheat malt, wheat bran and barley malt), wort 11 plateau When the average free ferulic acid concentration was compared, barley malt contained more ferulic acid than wheat malt, and only wheat bran had the highest concentration. Further, as shown in FIG. 11 (wheat bran addition amount and free ferulic acid production, average free ferulic acid concentration per 11 plateaus of wort (mg / L)), the ferulic acid production amount increased as the wheat bran addition amount increased. In order to increase the free ferulic acid content of beers, the free ferulic acid content can be increased by adding wheat bran to the barley malt.

It is a figure which shows the result of the comparison test of the free ferulic acid in the product beer of this invention, and the density | concentration of 4-VG. In the Example of this invention, it is a figure which shows the result in the test about the influence which the amount of enzymes has on the production | generation of free ferulic acid. In the Example of this invention, it is a figure which shows the result in the test about the production | generation of the kind of enzyme and free ferulic acid. In the Example of this invention, it is a figure which shows the result in the test about the production | generation of free ferulic acid with the enzyme single and combination of protease MG, laminex super, and cellulosin BGL. In the Example of this invention, it is a figure which shows the result in the test about the production | generation ratio of wheat malt and the free ferulic acid. It is a figure which shows the measurement result of the in vitro antioxidant activity of the ferulic acid high content beer in the Example of this invention. It is a figure which shows the measurement result of the blood antioxidant activity of the rat which orally administered the ferulic acid high content beer in the Example of this invention. It is a figure which shows the measurement result of the blood ferulic acid density | concentration of the rat which orally administered the ferulic acid high content beer in the Example of this invention. In the Example of this invention, it is a figure which shows the measurement result of the density | concentration of the free ferulic acid and 4-VG in the free ferulic acid high content baitzen beer which suppressed the baizen fragrance. In the Example of this invention, it is a figure which shows the average free ferulic acid density | concentration (mg / L) per 11 wort plateaus in the measurement test about the ferulic acid content of wheat, wheat malt, wheat bran, and barley malt. In the Example of this invention, it is a figure which shows the average free ferulic acid concentration (mg / L) per 11 plateaus of wort in the measurement test about wheat bran addition amount and free ferulic acid production.

Claims (9)

In the production of fermented alcoholic beverages using any one or more of barley, wheat, soybeans, barley malt and wheat malt as a brewing raw material, the saccharification step is performed using an enzyme having ferulic acid esterase activity, an enzyme having cellulase activity, and xylanase activity. In the presence of an enzyme having a saccharification temperature of 45 ° C. ± 10 ° C. and a saccharification temperature pattern with a protein resting time of 5 minutes or more, containing a high content of free ferulic acid and maintaining a flavor balance A method for producing a fermented alcoholic beverage. The enzyme having ferulic acid esterase activity is protease MG, the enzyme having cellulase activity is laminex super, and the enzyme having xylanase activity is cellulosin BGL. A method for producing a fermented alcoholic beverage containing and maintaining a flavor balance. The method for producing a fermented alcoholic beverage having a high content of free ferulic acid and maintaining a flavor balance, wherein wheat bran is added to the brewing material. The amount of free ferulic acid is 30 mg / L or more, The method for producing a fermented alcoholic beverage having a high content of free ferulic acid and maintaining a flavor balance according to any one of claims 1 to 3. The amount of enzyme added is increased until the amount of free ferulic acid in the wort reaches 30 mg / L, and the content of free ferulic acid according to any one of claims 1 to 4 is high, and the flavor balance The manufacturing method of the fermented alcoholic beverage which hold | maintained. The holding of flavor balance consists of suppressing the production | generation of 4-vinyl guaiacol in a fermentation process, The high content of the free ferulic acid of any one of Claims 1-5 characterized by the above-mentioned, and holding a flavor balance A method for producing a fermented alcoholic beverage. The production amount of 4-vinyl guaiacol is suppressed to 0.62 mg / L or less, and the production of a fermented alcoholic beverage containing a high content of free ferulic acid and maintaining a flavor balance according to claim 6 Method. The fermented alcohol having high free ferulic acid content according to any one of claims 1 to 7 and having a balanced flavor, wherein the fermented alcoholic beverage is beer, happoshu, or other brewed liquor. A method for producing a beverage. The method for producing a fermented alcoholic beverage having a high content of free ferulic acid and maintaining a flavor balance, characterized in that it is a baitzen beer in which the baiten flavor is suppressed and the content of free ferulic acid is high. .