JP4587368B2 - Fermented malt beverage containing lactic acid bacteria having antiallergic function or malt substitute fermented beverage, and method for producing the same - Google Patents

Description

  The present invention relates to a fermented malt beverage containing lactic acid bacteria having an antiallergic function or a malt substitute fermented beverage, and a method for producing the same. In particular, in the production of fermented malt beverages or malt substitute fermented beverages, by performing alcoholic fermentation with yeast after lactic acid fermentation with lactic acid bacteria having antiallergic activity, it has a refreshing refreshing acidity and mellow flavor, and anti The present invention relates to a fermented malt beverage containing lactic acid bacteria or malt substitute fermented beverages containing lactic acid bacteria having allergic activity, and a method for producing the same.

  Fermented malt beverages such as beer and happoshu are produced using malt as the main ingredient, starchy ingredients such as rice, wheat, corn, and starch as the auxiliary ingredients, and hops and water as raw ingredients. In the liquor tax law, the amount of malt used in beer is defined as 67% by weight or more in all raw materials when compared with malt use ratio. On the other hand, there are three types of Happoshu, the amount of malt excluding water, which is 50 wt% or more and less than 66.7 wt%, 25 wt% or more and less than 50 wt%, and less than 25 wt%. It is prescribed.

  Recently, malt that produces alcoholic beverages using a method similar to the production of fermented malt beverages such as beer and happoshu, using raw materials with a high nitrogen content and mineral content instead of malt in fermented malt beverages. Alternative fermented beverages are manufactured. Such malt-substitute fermented beverages are classified as “miscellaneous sake” under Japanese liquor tax law. According to the Japanese liquor tax law, “Zozake” is classified into “Happoshu” using malt or wheat as a part of raw materials and “Other miscellaneous drinks” other than Happoshu. Here, “other miscellaneous sake” does not use malt or wheat but mixes a nitrogen source obtained by enzymatic decomposition of plant proteins such as legumes and cereals with a saccharified solution and performs alcoholic fermentation with yeast.

  As mentioned above, Happoshu belongs to miscellaneous sake using malt as a raw material under Japanese liquor tax law. Both beer and Happoshu are derived from malt active enzymes and fungi. The saccharified malt and the auxiliary starch material are saccharified using a purified enzyme such as saccharified, and the saccharified solution is alcohol-fermented with yeast and decomposed into alcohol and carbon dioxide to produce an alcoholic beverage. Then it is common. Therefore, neither the method of making beer nor the method of making Happoshu has changed significantly. In addition, malt substitute fermented beverages belonging to “other miscellaneous sake” are also subjected to alcoholic fermentation with yeast by mixing a nitrogen source obtained by enzymatic decomposition of the above plant proteins and the saccharified solution instead of enzymatic decomposition of malt and the like. This is not basically different from the method of making beer or happoshu, but is produced in a similar manner using a beer or happoshu production apparatus.

  In recent years, in fermented malt beverages such as beer and happoshu, attempts have been made to develop fermented malt beverages having various flavors due to demands for diversification of consumer tastes. As an attempt to use microorganisms for diversification of the flavor, for example, in JP-A-6-303959, a raw material for producing happoshu such as malt and starchy auxiliary materials was prepared using yellow koji or white koji. A method for producing sake-flavored happoshu by adding sake brewer's sake and sake brewer's sake, as well as grape juice, apple juice and other fruit juice fermented and grown grapes as a raw material for happoshu Thus, a method for producing a sparkling liquor imparted with a wine flavor or a cider flavor is disclosed. Japanese Patent No. 2727048 discloses rice bran, barley koji, or corn koji obtained by breeding white koji mold or Rhizopus fungus in a saccharified solution prepared from a raw material for producing sparkling liquor such as malt and starchy auxiliary materials. A method for producing a sparkling liquor having a citric acid-rich flavor by adding a citric acid-rich source produced using

However, these are all methods for producing a sparkling liquor imparted with a sake flavor, a fruit liquor flavor, etc., which are different in flavor and type from conventional beer and sparkling liquor produced from malt and starchy auxiliary materials. It is a method for producing fermented malt beverages.
Also, since ancient times, in fermented malt beverages such as beer and sparkling wine, yeast is contained in the filtered fermented malt beverage, and as a so-called yeast-containing fermented malt beverage, flavorful and mellow fermentation Commercialization as a malt beverage has been carried out. However, since this yeast remains within the range of fermentation materials used in the production of fermented malt beverages such as beer and happoshu, there is a limit to the variety of flavor improvements.

  On the other hand, lactic acid bacteria are used as microorganisms for the production of alcoholic beverages in the field of sake and the like. However, in the production of fermented malt beverages such as beer and happoshu, rather, contamination is avoided as a contaminating bacterium. Therefore, in the production of fermented malt beverages such as beer and sparkling liquor, means for detecting it in order to avoid contamination with lactic acid bacteria (JP-A-7-289295, JP-A-9-121896, JP2003). -251 gazette, JP-T-2002-521650 gazette), means for removing it (JP-A-2002-315562, JP-A-2003-144128) and the like have been developed.

  On the other hand, in fermented malt beverages such as beer, a method is also known in which lactic acid bacteria are used and the acidity produced by lactic acid bacteria is used as a flavor. For example, Belgian lambic beer is known (Hideo Miyaji, “Beer Brewing Technology”, Food Industry Newspaper Co., Ltd. (December 28, 1999), P494-495). This lambic beer uses yeast and lactic acid bacteria mixed during brewing, and allows the microorganisms in a mixed state of the yeast and lactic acid bacteria to be fermented in parallel in a fermentation manner using a conventional beer fermenter. To manufacture. However, since this method uses lactic acid bacteria which happens to be mixed, there is a problem that it is difficult to control the concentration of the lactic acid bacteria, there is a problem in the stability of the taste, and the fermentation period is also long.

  Therefore, using one or more of sugar liquor, fruit juice, malt juice, and saccharified cereal as a fermentation raw material, the fermentation zone by yeast and the fermentation zone by acid-producing bacteria such as lactic acid bacteria are connected in series, and double fermentation is performed. A method has been developed for producing fermented beverages such as alcoholic beverages having a rich flavor or flavor with a refreshing sourness and ethanol produced by ethanol fermentation and a rich flavor or flavor (Japanese Patent No. 2888862). This method uses a refreshing sourness produced by lactic acid bacteria in a method for producing alcoholic beverages, etc., but when this method is applied to the production of fermented malt beverages such as beer, a fermentation process using lactic acid bacteria is required. In order to enter, it is necessary to control the diacetyl produced during fermentation.

  On the other hand, in recent years, in the field of foods and drinks and the like, food and drinks are actively used as functional foods and drinks by containing components such as physiologically active substances. One of its functions is antiallergic action. Lactic acid bacteria are known as one of the substances having this antiallergic action. Lactic acid bacteria used in the production of various foods and drinks are easy to apply to foods and drinks from the viewpoint of safety, and are useful materials as anti-allergic substances. However, when lactic acid bacteria are used for the production of functional foods and drinks as substances having anti-allergic action, even though lactic acid bacteria are generally referred to, not all lactic acid bacteria have a strong effect of enhancing anti-allergic action. When using, it is essential to select useful strains.

  To date, there are several known attempts to utilize lactic acid bacteria for anti-allergy. For example, the Lactobacillus rhamnosus LGG strain is known as an antiallergic lactic acid bacterium, and it has been shown that the onset of atopic dermatitis in children is suppressed by ingestion by pregnant women (Lancet, 2001, Vol. 357). 1076). Further, the use of lactic acid bacteria as an antiallergic agent is also disclosed in several patent publications. For example, Japanese Patent Laid-Open No. 9-2959 discloses L. Acidophilus, L. Brevis, L. Buchneri and L. Japanese Patent Application Laid-Open No. 10-309178 discloses the use of an anti-allergic agent having an IgE production amount of 30 ng / ml or less when mouse lymphocytes are cultured with lactic acid bacteria such as casei added. Use of bifidobacteria such as Infantis, Bifidobacterium breve, Bifidobacterium longum and Bifidobacterium bifidum as anti-allergic agents for treating food allergies, and No. 95697 discloses the use of lactic acid bacteria such as Enterococcus faecalis and Lactobacillus reuteri as inhibitors of type I allergies such as allergic bronchial asthma, allergic rhinitis and atopic dermatitis, respectively. .

  It is also known to use the antiallergic action of lactic acid bacteria for the production of food and drink. That is, as foods and drinks having an antiallergic effect, yogurt and lactic acid bacteria drinks using specific lactic acid bacteria as described above are partially put into practical use as foods and drinks having an antiallergic effect. However, such foods and drinks are from the field where the foods and drinks were originally manufactured using lactic acid bacteria, and the above-mentioned foods are selected from those having antiallergic activity for the lactic acid bacteria used in the production of the foods and drinks. In some cases, pursuing only the anti-allergic effect of lactic acid bacteria, there is another thing that does not satisfy in terms of taste, the use of anti-allergic activity of lactic acid bacteria, The characteristics of lactic acid fermentation and the antiallergic activity of lactic acid bacteria were not positively utilized in terms of their functions and taste.

  By the way, in the production of fermented malt beverages or malt substitute fermented beverages that have been alcoholic fermented with yeast as described above, using lactic acid bacteria that can be safely used as food and beverage materials, a unique taste with a refreshing acidity by lactic acid fermentation, It is conceivable to produce a fermented malt beverage or a malt substitute fermented beverage having excellent flavor and useful functionality by utilizing the antiallergic activity as described above. However, conventionally, in the production process of fermented malt beverages or malt substitute fermented beverages, it has functionality such as antiallergic activity due to problems such as diacetyl produced when lactic acid bacteria are added and lactic acid fermentation is performed. Moreover, there has been no fermented malt beverage or malt substitute fermented beverage containing lactic acid bacteria that can realize a harmonious, mild acidity and mild flavor.

JP-A-7-289295. Japanese Patent Laid-Open No. 9-2959. JP-A-9-121896. Japanese Patent Laid-Open No. 10-309178. JP 2000-95697 A. JP 2002-315562 A. Japanese Patent Laid-Open No. 2003-251. JP2003-144128A. JP-T-2002-521650. Japanese Patent No. 2888862. Hideo Miyaji “Beer Brewing Technology” Food Industry Newspaper Co., Ltd. (December 28, 1999), P494-495. Lancet, 2001, 357, 1076.

  An object of the present invention is to use a lactic acid fermentation by a lactic acid bacterium having antiallergic activity in a method for producing a fermented malt beverage or a malt substitute fermented beverage, having an antiallergic function, and having a refreshing refreshing acidity by lactic acid fermentation It is to provide a fermented malt beverage containing lactic acid bacteria or a malt substitute fermented beverage having a mellow flavor, and a method for producing the same.

  The present inventor has made extensive studies to solve the above problems, and in the manufacture of fermented malt beverages or malt substitute fermented beverages, after adding lactic acid bacteria having antiallergic activity and performing lactic acid fermentation, alcohol by yeast Alcohol fermentation by yeast with harmonious, refreshing and mild acidity derived from lactic acid fermentation by suppressing the diacetyl concentration of fermented malt beverage or malt substitute fermented beverage below the threshold of beer or happoshu It has been found that a fermented malt beverage containing lactic acid bacteria or a malt substitute fermented beverage having a mild flavor and flavor derived from lactic acid bacteria and having an antiallergic function due to the antiallergic activity of lactic acid bacteria can be produced, and the present invention has been completed.

  That is, in the present invention, in the production of fermented malt beverages and malt substitute fermented beverages, the fermented malt beverages and malt substitute fermented beverages are provided with an antiallergic function by using lactic acid bacteria having antiallergic activity, and the fermentation In the production process of malt beverages and malt substitute fermented beverages, after performing lactic acid fermentation with the lactic acid bacteria, by performing alcoholic fermentation with yeast, the acidity derived from lactic acid fermentation is compatible with the mild flavor and flavor derived from alcoholic fermentation Therefore, it is possible to achieve a harmonious, refreshing and mellow acidity, and further suppress the production of diacetyl by lactic acid fermentation as much as possible, and by performing alcoholic fermentation by yeast after the lactic acid fermentation, Diacetyl concentration is greatly reduced by the action of acetyl reducing ability, so that the concentration of diacetyl is the threshold of beer or happoshu. Fermented malt drink or malt substitute fermented drink with a refreshing sour taste with a refreshing sensation by reducing as much as possible the off-flavor components such as diacetyl, which is a typical off-flavor component, by suppressing to 0.1 mg / l or less It was found possible to manufacture the present invention, and the present invention was made.

  In the present invention, in order to suppress the production of diacetyl due to lactic acid fermentation, lactic acid fermentation is performed under anaerobic lactic acid fermentation control in an atmosphere in which an inert gas such as carbon dioxide or nitrogen gas is substituted. A method of suppressing the concentration below the threshold value of beer or happoshu can be employed. Moreover, in order to prevent the production | generation and residual of diacetyl as much as possible, the reduction | restoration of diacetyl in alcohol fermentation can be enlarged using top fermentation yeast as yeast in alcohol fermentation.

  In the present invention, as a lactic acid bacterium having antiallergic activity for imparting allergic function to fermented malt beverage or malt substitute fermented beverage, spleen derived from mouse spleen sensitized with ovalbumin by lactic acid bacteria is a medium containing ovalbumin When the test lactic acid bacterium is added to the suspension and cultured, Lactobacillus paracasei KW3110 strain is used as the test lactic acid bacterium, and the amount of interleukin 12 produced is 50% or more. Lactic acid bacteria exhibiting high anti-allergic activity and showing less than 50% of interleukin 4 production amount of Lactobacillus paracasei KW3110, Lactobacillus plantarum KW4110, Lactobacillus paracasei KW3925, Lactobacillus paracasei KW3926 strain or Streptococcu s salivarius KW3210.

  The present invention makes it possible to produce a fermented malt beverage or a malt substitute fermented beverage that does not cause a decrease in the antiallergic activity of lactic acid bacteria during long-term storage at room temperature. That is, the fermented malt beverage or malt substitute fermented beverage provided by the production method of the present invention is at room temperature, in the fermented malt beverage that has passed a period of 6 months or less corresponding to the market residence period after production, or the The production amount of the interleukin 12 and the production amount of the interleukin 4 indicated by the lactic acid bacteria in the malt alternative fermented beverage are within the range of the production amount of the interleukin 12 and the production of the interleukin 4 of the lactic acid bacteria. It is possible to provide a fermented malt beverage containing lactic acid bacteria or a malt substitute fermented beverage having an antiallergic function for preservation.

Specifically, the present invention relates to a method for producing a fermented malt beverage or a malt substitute fermented beverage, wherein a mouse spleen-derived lymphocyte sensitized with ovalbumin is suspended in an ovalbumin-containing medium in a fermentation raw material solution, and a test lactic acid bacterium When Lactobacillus paracasei KW3110 strain is used as a test lactic acid bacterium, the amount of interleukin 12 produced is 50% or more and less than 50% of the lactic acid bacterium-free control. Fermented malt by adding lactic acid bacteria showing high anti-allergic activity indicating the production amount of leukin 4 and performing lactic acid fermentation under lactic acid fermentation control under anaerobic conditions, followed by alcohol fermentation with yeast using top fermentation yeast diacetyl concentration of beverage or malt substitute fermented beverage suppressed below a threshold 0.1 mg / l of beer or malt liquors, and Anti fermentation malt beverage or malt substitute fermented beverage to a lactic acid 300~3000mg / L, fermented malt beverage or malt substitute fermented beverages per ^100ml, wherein the preparation to contain 109 ^{to 1011} pieces of lactic acid bacteria It comprises a method for producing a fermented malt beverage containing lactic acid bacteria having an allergic function or a malt substitute fermented beverage (Claim 1) .

The present invention, lactic acid bacteria showing high antiallergic activity that used in lactic acid fermentation, Lactobacillus paracasei KW3110 strain, Lactobacillus plantarum KW4110 strain, Lactobacillus paracasei KW3925 strain, or claim 1, wherein it is a Lactobacillus paracasei KW3926 strain A fermented malt beverage containing lactic acid bacteria having an antiallergic function or a method for producing a malt substitute fermented beverage ( Claim 2) .

  According to the method for producing a fermented malt beverage or a malt substitute fermented beverage of the present invention, it is a fermented malt beverage such as beer or sparkling liquor or a malt substitute fermented beverage, which provides a refreshing and refreshing mellow acidity and a mild flavor. In addition, by the coexistence of lactic acid bacteria having antiallergic activity, a fermented malt beverage or a malt substitute fermented beverage having antiallergic functionality can be produced. Furthermore, the method for producing a fermented malt beverage or a malt substitute fermented beverage of the present invention can provide a fermented malt beverage or a malt substitute fermented beverage that does not cause a decrease in the antiallergic activity of lactic acid bacteria with respect to long-term storage at room temperature.

Moreover, according to the method for producing a fermented malt beverage or a malt substitute fermented beverage of the present invention, a fermented malt beverage or a malt substitute fermented beverage obtained by lactic acid fermentation and alcohol fermentation by yeast is separately produced, and lactic acid fermentation is performed. It is possible to prepare a fermented malt beverage or a malt substitute fermented beverage produced by a method for producing a normal fermented malt beverage or a malt substitute fermented beverage, and to adjust the flavor to the target. At that time, in order to obtain a final amount of lactic acid bacteria of 10 ⁹ to 10 ¹¹ per 100 ml, it is also possible to adjust by adding the same bacteria that are separately cultured and generated.

  The present invention relates to a method for producing a fermented malt beverage or a malt substitute fermented beverage. After adding lactic acid bacteria having antiallergic activity and performing lactic acid fermentation, alcohol fermentation with yeast is performed, and fermented malt beverage or malt substitute ferment By controlling the diacetyl concentration of the beverage to be below the threshold of 0.1 mg / l for beer or happoshu, a harmonious, refreshing and mellow acidity derived from lactic acid fermentation and a mild flavor and flavor derived from yeast alcohol fermentation And producing a fermented malt beverage containing lactic acid bacteria or a malt substitute fermented beverage having an antiallergic function due to the antiallergic activity of lactic acid bacteria.

  In the method for producing a fermented malt beverage or a malt substitute fermented beverage of the present invention, the lactic acid bacteria used for lactic acid fermentation are lactic acid bacteria having antiallergic activity, and the lactic acid bacteria include mouse spleen-derived lymphocytes sensitized with ovalbumin. When suspended in an ovalbumin-containing medium and added with a test lactic acid bacterium and cultured, Lactobacillus paracasei KW3110 strain is used as the test lactic acid bacterium, showing an interleukin 12 production amount of 50% or more, and The lactic acid bacteria which show the high antiallergic activity which show the production amount of interleukin 4 of less than 50% of lactic acid bacteria addition control can be mentioned. Moreover, it is preferable that it is a lactic acid bacterium which can maintain the activity for a long period of time. Specific examples of the lactic acid bacteria include Lactobacillus paracasei KW3110 strain, Lactobacillus plantarum KW4110 strain, Lactobacillus paracasei KW3925 strain, Lactobacillus paracasei KW3926 strain or Streptococcus salivarius KW3210.

  In the present invention, in order to obtain a lactic acid bacterium having high antiallergic activity used for lactic acid fermentation, animal spleen-derived lymphocytes sensitized with allergen are suspended in the allergen-containing medium, added with a test lactic acid bacterium, and cultured. 50% or more of Th1-induced cytokine production as compared to the case of using Lactobacillus paracasei KW3110 as a test lactic acid bacterium or 50% or more compared to the case of using L. paracasei KW3110 A lactic acid bacterium having an interleukin 4 production amount of less than% is isolated to obtain a lactic acid bacterium having high antiallergic activity.

  That is, using a known lactic acid bacterium strain or a newly isolated lactic acid bacterium strain, mouse spleen-derived lymphocytes sensitized with ovalbumin were suspended in an ovalbumin-containing medium and cultured, Basically, L. as a test lactic acid bacterium. Equivalent to using paracasei KW3110 strain or L. Acquired by isolating lactic acid bacteria that show 50% or more interleukin 12 production and less than 50% interleukin 4 production of the lactic acid bacteria-free control compared to when paracasei KW3110 strain is used be able to. As described above, the lactic acid bacterium used as an active ingredient in the present invention is based on the index of the present invention by carrying out an experimental method for evaluating the antiallergic activity in vitro among various known or newly isolated lactic acid bacteria. A person skilled in the art can easily select and obtain the data.

  In the present invention, lactic acid bacteria having high antiallergic activity used for lactic acid fermentation are exemplified by L. The paracasei KW3110 strain can be obtained from the Japan Dairy Industry Association as the L.casei L14 strain. According to the description of the Japan Dairy Technology Association, Although there is a description of casei, when the present inventors analyzed using RFLP (Restriction Fragment Length Polymorphism) and AFLP (Amplified Fragment Length Polymorphism) using a QUALICON riboprinter, the strain was found to be L. casei. Since it was determined to be paracasei, L. Indicated as paracasei. L. used as an active ingredient of the antiallergic composition in the present invention. paracasei KW3110 can be obtained from the Japan Dairy Industry Association as described above, but based on the Budapest Treaty for the Deposit of Patent Microorganisms, the International Depository Agency, National Institute of Advanced Industrial Science and Technology, Patent Biological Deposit Center Has been deposited as FERM BP-08634.

  In addition, L. lactic acid bacteria having high antiallergic activity used for lactic acid fermentation in the present invention are used. plantarum KW4110 strain plantarum JCM1149 strain, The paracasei KW3926 strain Paracasei JCM8132 strain can be obtained from JCM (RIKEN Microbial System Storage Facility). paracasei KW3925 strain It can be obtained from NRIC (Tokyo University of Agriculture) as paracasei NRIC1917.

  In the method for producing the fermented malt beverage or malt substitute fermented beverage of the present invention, lactic acid fermentation is performed using the lactic acid bacteria having the antiallergic activity, and then alcohol fermentation with yeast is performed. In the method for producing a fermented malt beverage or malt substitute fermented beverage of the present invention, as a fermentation raw material that can be assimilated by lactic acid bacteria and yeast, saccharification using assimilation sugars, fruit juice, wort, and grains containing oligosaccharides as raw materials A fermentation raw material liquid mainly composed of one or more of the liquids can be used. In particular, in the case of fermented malt beverages such as beer and happoshu, wort is used. The process currently performed is employ | adopted for the preparation process in the case of wort manufacture. That is, the process normally performed for manufacture of fermented malt drinks, such as beer and happoshu, or malt substitute fermented drinks is employ | adopted. However, it is desirable that the wort used for lactic acid fermentation does not add hops having a bacteriostatic action against many lactic acid bacteria. In order to make a beer or a sparkling liquor with a refreshing taste of hops, it is desirable to add yeast wort with hops after lactic acid fermentation, and to add isomerized hops before filling.

  As conditions for lactic acid fermentation, basically, fermentation conditions for normal lactic acid fermentation can be employed. That is, in order to perform an appropriate lactic acid fermentation, a temperature condition suitable for the bacterium used is adopted, and the pH is controlled during the fermentation so that the lactic acid fermentation is not inhibited by the pH reduction due to the lactic acid produced by the bacterium itself. It is desirable.

In the method for producing a fermented malt beverage or a malt substitute fermented beverage of the present invention, it is important to suppress the diacetyl concentration of the fermented malt beverage or the malt substitute fermented beverage to a threshold value of 0.1 mg / l or less for beer or happoshu. . For that purpose, it is necessary to reduce as much as possible the concentration of diacetyl produced in lactic acid fermentation by lactic acid bacteria having an antiallergic function. As the method, the production of diacetyl is suppressed by lactic acid fermentation management under anaerobic conditions. For example, at the time of lactic acid fermentation, an atmosphere substituted with an inert gas such as CO ₂ , N ₂ or a rare gas is created, and the formation of diacetyl after lactic acid fermentation is minimized by making it an highly anaerobic condition. Can be suppressed.

  In the method for producing a fermented malt beverage or a malt substitute fermented beverage of the present invention, as a method of reducing the diacetyl concentration, the yeast used for alcoholic fermentation is top fermented yeast, so that diacetyl produced by lactic acid fermentation is finally obtained. It is also possible to reduce to below the threshold. It is known that the top fermenting yeast is fermented at a higher temperature than the bottom fermenting yeast, and therefore has a high diacetyl reducing ability.

  In the method for producing a fermented malt beverage or a malt substitute fermented beverage of the present invention, alcoholic fermentation with yeast is performed after lactic acid fermentation with lactic acid bacteria. Alcohol fermentation by this yeast enables the sourness derived from lactic acid fermentation to be combined with the mild flavor and flavor derived from alcoholic fermentation to create a harmonious, refreshing and mellow sourness. By greatly reducing diacetyl due to the action of the acetyl reducing ability, the diacetyl concentration can be suppressed to a threshold of 0.1 mg / l or less for beer or happoshu. Alcohol fermentation by yeast after lactic acid fermentation can employ fermentation conditions generally employed in the production of fermented malt beverages or malt substitute fermented beverages. That is, the process normally performed in manufacture of fermented malt drinks, such as beer and sparkling liquor, or malt substitute fermented drinks can be employ | adopted.

The fermented liquor after the alcoholic fermentation by yeast is appropriately prepared separately, and is prepared by a fermented malt beverage or a malt substitute fermented beverage produced by a normal fermented malt beverage or malt substitute fermented beverage that is not subjected to lactic acid fermentation. However, it is possible to adjust the target flavor. In that case, if it mixes after a filtration process, since quality control will become comparatively easy, it is more preferable. At that time, in order to make the final amount of lactic acid bacteria to 10 ⁹ to 10 ¹¹ per 100 ml, the same bacteria separately cultured by a general lactic acid bacteria culture method should be added and adjusted before the filling step. Is possible.

In order to make the fermented malt beverage or the malt substitute fermented beverage of the present invention beer or happoshu having a harmonious, mild acidity and mild flavor, the lactic acid content of the final product is 300 to 3000 mg / l, and It is preferable to adjust so that the pH in that case does not fall below 3.8. At the same time, the concentration of lactic acid bacteria having antiallergic activity is desirably 10 ⁹ to 10 ¹¹ lactic acid bacteria per 100 ml of fermented malt beverage or malt substitute fermented beverage. Furthermore, it is important that the diacetyl concentration be 0.1 mg / l or less than the threshold of beer or happoshu. The antiallergic activity by the lactic acid bacteria of the fermented malt beverage or the malt substitute fermented beverage produced in the present invention is that the fermented malt beverage or the malt substitute fermented beverage has passed through an average market residence period, that is, 6 Even if stored for months, it does not change.

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

1 w / w% of lactic acid bacteria Lactobacilllus paracasei KW3110 strain pre-cultured in the same wort for 24 hours at 32 ° C. was added to the hopless wort adjusted to a sugar content of 14 ° P., and lactic acid fermentation was performed at 32 ° C. (The initial number of lactic acid bacteria is 10 ⁶ cells / ml). The lactic acid fermentation tank had a capacity of 20 L, the stirring speed was 10 rpm, and the inside of the tank was replaced with nitrogen gas. The tank was set to a slightly positive pressure and was not vented. Fermentation was carried out for 48 hours, and sodium hydroxide was added so that the pH did not drop below 4.8 until 24 hours. The fermentation broth was sterilized by a plate heat exchanger of 80 ° C. × 30 seconds. The obtained fermented broth contained 8.0 × 10 ¹⁰ Lactobacilllus paracasei KW3110 strains per 100 ml, and although it was somewhat sweet, it had a very refreshing and mellow acidity and a sweet taste of malt juice. The composition of the lactic acid fermentation broth was as shown in Table 1. The number of bacteria after sterilization was measured by a method using real-time PCR. Specifically, the sample was centrifuged (15000 rpm, room temperature, 5 minutes), the precipitate was suspended in TE buffer, glass beads (50-212 microns, SIGMA) were then added to the cell suspension, and the vortex mixer was added. The tube was set upside down on (Model Vortex-Genie 2), vigorously stirred for 2 minutes, the cells were crushed, and centrifuged (15000 rpm, room temperature, 5 minutes), and the supernatant was used as a DNA sample. On the other hand, Roche LightCycler was used as a real-time PCR instrument, and FastStart DNA Master SYBR Green I was used for the reaction. MgCl ₂ concentration is 3 mM, and L. paracasei detection primer
PARA; 5'-CACCGAGATTCAACATGG-3 '(SEQ ID NO: 1)
Y2; 5′-CCCACTGCTGCCTCCCGTAGGAGT-3 ′ (SEQ ID NO: 2)
It was used.

Add the same amount of hopped wort adjusted to 14 ° P sugar content, add 0.6 w / w% brewer's yeast (Saccharomyses cerevisiae), ferment at 10 ° C for 1 week, and store for about 1 month. Then, filling was performed after adjusting the alcohol to 5.5 v / v%. This beer contains 2.5 × 10 ¹⁰ cells of KW3110 strain per 100 ml
/ Ml, and the flavor had a taste that harmonized the soft sourness and the refreshing feeling peculiar to the malt fermented beverage. The composition of this malt fermented beverage was as shown in Table 2.

(Evaluation of antiallergic ability of lactic acid bacteria in vitro)
1. Sample A part of each fermentation broth for measuring antiallergic activity was centrifuged at 9000 G, and the resulting precipitate was collected and suspended in sterilized water, and then centrifuged at 500 to 1000 G. Thereafter, the supernatant was centrifuged at 3000 G, and the resulting precipitate was lyophilized and suspended in PBS.

2. Experimental animals, rearing 7-10 week old BALB / C mice (Charles River) were intraperitoneally injected with day 0 and day 6 with 1 mg of ovalbumin (OVA) together with 2 mg of adjuvant Alum. Day 13 was dissected and the spleen was isolated and lymphocytes were prepared.

3. Measurement of splenocytes IL-4 and IL-12 IL-4 and IL-12 were measured using 0ptEIA ELISA Set (Becton Dickinson).

4). Measurement conditions Spleen lymphocytes prepared by the method of “2. Experimental animals, rearing” were suspended in RPMI 1640 (SIGMA) + 10% FCS (Roche) +1 mg OVA medium to 2.5 × 10 ⁶ cells / ml. Further, lactic acid bacteria to be tested were added at 0.1 or 1 μg / ml. One week later, the culture supernatant was collected, and IL-12 was measured as Th1 cytokine and IL-4 was measured as Th2 cytokine.

5). Experimental Results After lactic acid fermentation, then yeast fermentation, lactic acid bacteria contained in the malt fermented beverage product were sensitized with ovalbumin and mouse spleen-derived lymphocytes were suspended in ovalbumin-containing medium and the test lactic acid bacteria were added. As a result of culturing, as shown in FIGS. 1 and 2, compared to the case where L. paracasei KW3110 strain was used as the test lactic acid bacterium, the production amount of interleukin 12 was 50% or more and 50 of the lactic acid bacterium-free control was obtained. The amount of interleukin 4 produced was less than%, and the amount was almost unchanged even after yeast fermentation (control was measured without addition of lactic acid bacteria).

    In Example 1, in order to adjust the fermented malt beverage that had been fermented and stored to a malt fermented beverage with less sourness characteristics, alcohol 4.3% / v%, appearance extract 1.1 ° P filtered Filling was performed after blending 25% with a malt fermented beverage. The flavor had a light and refreshing taste with a soft sour taste. The composition of this malt fermented beverage was as shown in Table 3.

  As a result of suspending mouse spleen-derived lymphocytes sensitized with ovalbumin in lactic acid bacteria contained in the fermented malt beverage product in an ovalbumin-containing medium and adding the test lactic acid bacteria, the final product of Example 1 The production amounts of equivalent interleukin 12 (IL-12) and interleukin 4 (IL-4) are shown.

  Changes in production of IL-12 and IL-4 over time when the malt fermented beverage product is stored at room temperature of 20 to 30 ° C. for 6 months are shown in FIGS. As a result, it was confirmed that almost no change over time in the production amount of IL-12 and IL-4 was observed, and both maintained high activity.

In Examples of the present invention, after lactic acid fermentation and then yeast fermentation, lactic acid bacteria contained in a malt fermented beverage product were sensitized with ovalbumin and mouse spleen-derived lymphocytes were suspended in ovalbumin-containing medium and tested. It is a figure which shows the production amount of IL-12 as a result of adding and cultivating lactic acid bacteria. In Examples of the present invention, after lactic acid fermentation and then yeast fermentation, lactic acid bacteria contained in a malt fermented beverage product were sensitized with ovalbumin and mouse spleen-derived lymphocytes were suspended in ovalbumin-containing medium and tested. It is a figure which shows the production amount of IL-4 as a result of adding and culture | cultivating lactic acid bacteria. In the Example of this invention, it is a figure which shows the production amount of IL-12 when the malt fermented drink product of this invention is preserve | saved at room temperature of 20-30 degreeC for 6 months. In the Example of this invention, it is a figure which shows the production amount of IL-4 when the malt fermented drink product of this invention is preserve | saved at room temperature of 20-30 degreeC for 6 months.

Claims (2)

In the production method of fermented malt beverage or malt substitute fermented beverage, when cultivated by suspending mouse spleen-derived lymphocytes sensitized with ovalbumin in ovalbumin-containing medium and adding test lactic acid bacteria to the fermentation raw material solution, Compared to the case where Lactobacillus paracasei KW3110 strain (deposit number: FERM BP-08634) is used as the test lactic acid bacterium, the production amount of interleukin 12 is 50% or more and less than 50% of the control with no lactic acid bacterium added. Fermented malt by adding lactic acid bacteria showing high anti-allergic activity indicating the production amount of leukin 4 and performing lactic acid fermentation under lactic acid fermentation control under anaerobic conditions, followed by alcohol fermentation with yeast using top fermentation yeast The diacetyl concentration of beverages or malt substitute fermented beverages is controlled to be below the threshold of 0.1 mg / l for beer or happoshu. And fermented malt beverage or malt substitute fermented beverage contains 300 to 3000 mg / L lactic acid and 10 ⁹ to 10 ¹¹ lactic acid bacteria per 100 ml fermented malt beverage or malt substitute fermented beverage A method for producing a fermented malt beverage containing lactic acid bacteria or a malt substitute fermented beverage having an antiallergic function. The lactic acid bacterium exhibiting high antiallergic activity used for lactic acid fermentation is Lactobacillus paracasei KW3110 strain (deposit number: FERM BP-08634) , Lactobacillus plantarum KW4110 strain, Lactobacillus paracasei KW3925 strain, or Lactobacillus paracasei KW3926 strain. Item 2. A method for producing a fermented malt beverage containing lactic acid bacteria having an antiallergic function according to Item 1 or a fermented malt substitute beverage.