JP3362310B2 - Method for producing alcoholic beverage and alcoholic beverage obtained thereby - Google Patents

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for producing alcoholic beverages such as sake, beer, wine and the like using a basidiomycete having amylase activity and alcohol dehydrogenase activity, and an alcoholic beverage obtained thereby. .

[0002]

2. Description of the Related Art Conventionally, alcoholic beverages such as sake, beer and wine have been produced by alcoholic fermentation of yeast.

Conventionally, in the production of sake, koji mold has been used for the purpose of saccharifying starch and yeast has been used for the purpose of alcohol fermentation. Sake is generally produced by alcoholic fermentation of yeast with yeast while saccharifying steamed rice with Koji mold. For example, it is manufactured according to the flow sheet shown in FIG. That is, first add koji and water to steamed rice,
Yeast is grown on this. This is called liquor, and when liquor is formed, steamed rice, koji and water are added 3 times and fermented for about 1 month. During this time, saccharification of starch and alcoholic fermentation are performed. Then, by squeezing, orienting, and sterilizing by burning, the aged moromi that has completed fermentation
You can get sake.

[0004] Beer is generally obtained by germinating barley to form malt, saccharifying barley starch, and then alcohol-fermenting wort containing hops. For example, in FIG.
It is manufactured according to the flow sheet shown in. That is, first, barley is soaked in water to absorb water, and then 1
Germinate at 3 to 17 ° C for about 1 week. This is called green malt, which is produced by enzymes and is in a state suitable for saccharification.
Next, the malt is heated and dried to generate a unique color and aroma, and then crushed. After that, auxiliary materials such as rice and corn starch are added to saccharify the starch, hops are added to the filtered product, which is boiled and filtered to make wort. afterwards,
Cool the wort and add yeast to ferment. In the case of fermentation by top yeast, it is aged at 15 to 20 ° C. for 4 to 6 days, and in the case of fermentation by bottom yeast, it is aged at 0 to 2 ° C. for 1 to 3 months to dissolve carbon dioxide in beer. Then, when clear beer is directly packed in a bottle or a can, draft beer is obtained, and when heat sterilized, ordinary beer is obtained.

Wine is generally obtained by fermenting grape juice with yeast, and is roughly classified into red wine and white wine. Red wine is manufactured, for example, according to the flow sheet shown in FIG. 7. That is, first, the grapes of dark red or black-purple varieties are crushed, and fruit juice, peel,
Collect the seeds. Then, a sulfite compound (potassium metabisulfite, etc.) is added for the purpose of preventing the growth and oxidation of various bacteria, and further sugar is adjusted with sugar, glucose or the like to adjust the sugar content, and then yeast is added to add 20 to 25. Ferment at 7 ° C for 7-10 days. After that, squeezing, post-fermentation, pulling, barrel storage,
Red wine is obtained by filling each in a container such as a bottle through each step of filtration. In addition, white wine is produced in substantially the same manner as red wine, but only fruit juice is fermented with yeast using grapes of green-yellow varieties as a raw material, and usually after fermenting at 15 ° C for about 3 weeks. It is obtained by squeezing, stuffing in a barrel, and aging.

[0006]

As described above, alcoholic beverages have been conventionally produced using yeast having an alcohol fermenting ability, but there have been few reports of production using bacteria other than yeast. It is no exaggeration to say that there are no reports of bacteria. Moreover, since yeast has a small alcohol fermentation ability under aerobic conditions, there are restrictions on the production conditions for alcoholic beverages. Furthermore, the alcoholic beverage produced using yeast hardly contains β-D-glucan, which is an anticancer substance, and cannot be expected to have a thrombosis preventive effect such as myocardial infarction and cerebral thrombosis.

The present invention has been made in view of such circumstances, and in the alcohol fermentation step, instead of yeast,
It is an object of the present invention to provide a completely new method for producing an alcoholic beverage using basidiomycetes and an alcoholic beverage obtained thereby.

[0008]

In order to achieve the above object, the present invention is a method for producing an alcoholic beverage in which alcohol fermentation is carried out by a bacterium having alcohol fermentation ability, wherein the bacterium having alcohol fermentation ability is amylase. The first gist is a method for producing an alcoholic beverage which is a basidiomycete having activity and alcohol dehydrogenase activity.

A second aspect of the present invention is an alcoholic beverage obtained by the above method for producing an alcoholic beverage.

The present inventors have conducted various experimental studies on basidiomycetes which produce useful physiologically active substances. In the process, basidiomycetes were conventionally considered to have no alcohol fermenting ability, but it was found that among various types of basidiomycetes, ones having amylase activity and alcohol dehydrogenase activity exist. Then, as a result of trying to produce various alcoholic beverages using such basidiomycetes, it was found that various alcoholic beverages having the same alcohol content as conventional ones were obtained, and the present invention was reached.

Using the above-mentioned basidiomycetes, alcoholic beverages can be produced under anaerobic conditions, and alcoholic beverages can be produced even under aerobic conditions, as in the conventional production of various alcoholic beverages. it can. Therefore, there is an advantage that the production conditions of alcoholic beverages can be relaxed.
Further, in the case of anaerobic conditions, there is an advantage that the conventional manufacturing equipment can be used as it is and the alcoholic beverage can be manufactured without adding new equipment.

Since the alcoholic beverage of the present invention is produced by a specific basidiomycete, not by the conventionally used yeast, it contains a useful physiologically active substance which cannot be produced by yeast. That is, β-D-glucan that exhibits antitumor properties, immunoreactivity, antiallergic properties, and dietary fiber effects, chitin that exhibits dietary fiber effects, and heteropolysaccharide [pectin, hemicellulose, active hemicellulose complex (AHCC, Active hemicellulo
se complex), polyuronide, lignin] and the like. Therefore, the alcoholic beverage of the present invention becomes a novel beverage which has never existed before and can be a functional and healthy beverage.

In the present invention, the basidiomycete means that the fruiting body is referred to as a "mushroom", and in addition to basidiomycetes in the taxonomic classification of microorganisms (Agaricales, Hydrangea mushrooms, Abdominal fungi, Fungus fungi). , The concept also includes some of the ascomycetes.

In the present invention, the anaerobic condition means
It means the condition that the oxygen partial pressure is less than 1.0 kPa, and also includes the condition where there is no oxygen. On the other hand, the aerobic condition means a condition that the oxygen partial pressure is 1.0 kPa or more.

[0015]

BEST MODE FOR CARRYING OUT THE INVENTION Next, embodiments of the present invention will be described.

The method for producing an alcoholic beverage of the present invention is a method for producing various conventionally known alcoholic beverages in which alcohol fermentation is carried out by a bacterium having an alcohol fermentation ability, and a specific basidiomycete is used as the bacterium having an alcohol fermentation ability. Alcohol fermentation is performed by basidiomycetes.

The alcoholic beverages targeted by the present invention include brewed alcohol such as sake, beer and wine, as well as distilled alcohol such as shochu, whiskey and brandy.

The specific basidiomycete used in the present invention is not particularly limited as long as it has amylase activity and alcohol dehydrogenase activity. For example, Agaricus blazei, collected in forests in various places, Agaricus blazei, Pleurotus ostreatus
, Plerotos Ostretas), Enoki mushroom (Flammul)
ina velutipes, Flamlina velutipes, Tricholoma matsutake, Astericularia auricula, Alicicularia auricularia, Kuritake (Naematoloma sublateritium, Naematoloma sublateritium), Shiitake (Lentinus)
dodes, Lentinas Eddes), Shoro (Rhizopog)
on rubescens, Rhizopogon rubesense), Suehirotake (Schizophyllum commune, Schizophyllum commune), Pleurotus citrinopileatus, Pleurotus citrinopileatus, Pleurotus citrinopiletas, Deux maitake (De)
ndropolyporus umbellatus, dendropoliporus uberatas), bukuryou (Poriacocos, polyacokos), beech shimeji (Hypsizygus marmoreus, hypsizygas marmores), maitake (Grifola frandosa, glyphora frandosa), mustache (Laetiporus sulphureus)
s, Laetipora sulfures), Ganoderma lucidum (Gano)
derma lucidum, Ganoderma lucidum), Agaricus bisporus (Agaricus bisporus, Agaricus bisporus), willow matsutake (Agrocybe cylindracea, Aguroshibe cylindracea), mountain blanking Shitake (Hericium erinaceum,
Helicium Erinasem, Coriolus versi
color, Coriolus vyazykaraa), Elfvingia applanata (Elfingia aplanata), Nameko (Pholiota nameco, Poliota nameko), Plumiota adiposa (Pholiota adiposa)
Adiposa) and the like.

As basidiomycetes used for the production of sake in alcoholic beverages, in addition to the above-mentioned basidiomycetes, morel mushrooms (Morchella esculenta, morsella esculenta), ximeji (Tricholoma flavovireus, tricoloroma flavovires), white hatsu (Russula delica) ，
Rushla Delica), Tsugasaru no Shikae (Fomitopsis)
pinicola, Phymitosis pinicolas), mushroom (Ca)
lvatia craniiformis, Caradya claniiformis, Cordyceps dipterigene, Cordaria crabiformis, Ramaria bo
trytis, La Maria Botrytis), Pleurotus ostreatus (Tric
holoma robustum, Torikoromabustan), Murasashimeji (Lepista nuda, Lepista nuda), Yamaguchi (Leccinum scabrum, Reshinamsukaburan), black hatu (Russula nigricans, Rusula nigricans), Hatakeshijime (Lyophyllum decastes, Lyophyllum decastes) Boletus edulis, Boletus edulis) and the like.

As basidiomycetes used for the production of beer in alcoholic beverages, in addition to the above-mentioned basidiomycetes, Auricularia polytricha (Auricularia polytricha) and white shimejime (Tricholoma japonicu)
m, Tricoloroma japonicum), Pleurotus cornucopia (Lactarius)
volemus, Lactarias borems), squirrel (Suillus luteus, Suirus luteus), Lactarius hatsudake, Lactarius hatusudake,
Oyster mushrooms (Spathularia flavida, spatula flavida), pine trees (Lentinus lepideus, lentinas repi), musk mushrooms (Panellus serotinus, pannelus serotonas), yabrebenitatake (Russula rosacea)
, Russula rosacea), Ganoderma lucidum (Fistulina hap
atica, phytulina hepatica), dragonfly (Ganod)
erma neo-japonicum, Ganodermaneo japonicum), Hanabitake mushroom (Sparassis crispa, Sparassis)
Crispa), Pleurotus cornucopia (Cordyceps militaris), and the like.

As the basidiomycetes used for the production of wine in alcoholic drinks, in addition to the above-mentioned basidiomycetes, Amytake mushrooms (Suillus bovinus, Syrus bovinus), Tremella fuciformis, Phallus impudicus ， False Impdecas） 、 Nishawaharitake (Pseudohydnum)
gelatinosum, Sudhidonum Gelatinosam), Novoriryu (Helvella crispa, Helvera crispa), Beech mushroom (Mycoleptodonoides aitchisonii)
, Mycoreptodonoides aichisonii), Lyophyllum shimeji, Lyophyllum shimeji,
Pleurotus cornucopiae (Bankera fuligineo-alba, Bankera)
Figineo Alba), Molinocaretake (Collybia d)
ryophila, Colivia doriophylla), Tamachoreitake (Polyporus tuberaster),
Ginger (Rozites caperatus, Logites caperatas), Pleurocybella porrigens (Pleurocybella porrigens, Pleurocybera porgens), Shaka shimeji (Lyophyllum)
fumosum, Ryofilam fumosum) and the like.

Amira in the above specific basidiomycete
The enzyme activity can be confirmed, for example, as follows.
it can. That is, first, soluble starch 2.5% by weight
50 mM Acetate buffer containing 5 mM CaCl _{2 in} which is dissolved
Is prepared and used as a substrate solution. Next, the substrate solution 3
Add 100 μl of crude enzyme solution to 00 μl, and for 5 minutes at 40 ° C.
React. Enzyme reaction is heated in boiling water for 10 minutes
Stop. After stopping the reaction, add 1.0 ml of 0.5 to the reaction solution.
Add 3.0 ml of N-acetic acid and 0.1% iodine solution and stir well
And then using a double beam spectrophotometer to absorb at 700 nm
Degrees were measured, this value is an initial value (blank, the crude enzyme solution
If the value is less than the value measured using water instead,
It can be said to have a millase activity. In addition, first
The above amylase activity was confirmed even in the listed basidiomycetes
Those which are not used cannot be used in the present invention.
Yes.

Alcohol in the above specific basidiomycete
The presence or absence of dehydrogenase activity can be determined, for example, as follows.
You can check. That is, first, for basidiomycetes
Ultrasonication and centrifugation are performed, and the supernatant is used as the crude enzyme solution.
And prepare. In addition, 0.1 M Tris buffer (pH 7.
4) 12.5 ml , NAD ⁺ 0.1 ml, 0.1M
11.0 ml of ethanol and phenazine methsulfate
(PMS) 0.5 ml and nitro blue tetrazoliu
0.5 ml (NBT) and 10 ml of water are mixed and mixed.
A reaction solution is prepared by combining. Next, above
Using 0.02 ml of crude enzyme solution, polyacrylamide gel
After electrophoresis, run the polyacrylamide gel on top.
Immerse in the above reaction solution and react at 30 ° C. for 1 hour. That
Then, the stained alcohol dehydrogenase band can be confirmed.
If it comes, it means that it has alcohol dehydrogenase activity.
I can say. Even if the basidiomycetes listed above are used,
If the rucor dehydrogenase activity is not confirmed, the present invention
Cannot be used in.

Among the above-mentioned series of basidiomycetes, one having an alcohol dehydrogenase activity of 1.0 units / mg or more is preferable, and one having 10.0 units / mg or more is most preferable. That is, if the alcohol dehydrogenase activity is too low, it is necessary to take measures such as using a large amount of basidiomycetes, which is not preferable in the production of alcoholic beverages. The alcohol dehydrogenase activity was determined by NADH using a supernatant obtained by subjecting basidiomycetes to ultrasonic treatment and centrifugation as a crude enzyme solution, using ethanol, Tris buffer, NAD ^+, etc.
It is determined by measuring the absorbance at 0 nm.

Among the above-mentioned basidiomycetes, enokitake, matsutake mushroom, mustache and tsukuritake are preferable for producing beer in consideration of availability, alcohol dehydrogenase activity and the like, and also for producing wine. In view of availability, alcohol dehydrogenase activity, etc., agaricus mushrooms, enokitake mushrooms, oyster mushrooms, oyster mushrooms are suitable, and for the production of sake, availability, alcohol dehydrogenase activity, In consideration of the amylase activity and the like, Agaricus cystate, Matsutake mushroom, and Buna shimeji are preferable. In particular,
Tsukuritake is suitable for the production of beer in terms of its refreshing taste and aroma, Ganoderma lucidum is suitable for the production of wine in terms of its fruity taste and aroma, and beech syrup is said to have a rich taste. In terms of aroma, it is suitable for the production of sake.

The method for producing an alcoholic beverage of the present invention is carried out, for example, according to the flow sheet shown in FIG. That is, first, prepare the raw material, add the above basidiomycete to it,
An alcoholic beverage is manufactured by performing alcoholic fermentation under aerobic conditions.

Regarding the method for producing the alcoholic beverage of the present invention,
Taking sake as an example, description will be given along the flow sheet shown in FIG. That is, first, 1 to 10 of the above basidiomycetes is added to steamed rice.
Kokokoji mushrooms (grown with basidiomycetes), which have been added by about 5% by weight, are mixed with steamed rice and water to grow the basidiomycetes, thereby producing sake mothers. As the basidiomycete, it is preferable to use a pure culture that is liquid-cultured and grown as a mycelium. Next, steamed rice, mushroom mushrooms, and water are added to the liquor mother three times, and the fermentation is performed under aerobic conditions for about one month. During this period, saccharification of starch by the basidiomycete (saccharification step) and alcoholic fermentation (fermentation step) are performed. That is, a single fermentation is performed instead of the conventional parallel multiple fermentation. After that, as in the conventional sake production process, the fermented moromi mash is squeezed, squeezed,
By passing through each step of sterilization by burning and filling in a container such as a bottle, sake can be obtained in which saccharification of starch and alcohol fermentation are performed only by the basidiomycete.

Also, beer will be described as an example along the flow sheet shown in FIG. That is, first, barley is immersed in water to absorb water, and then germinated at 13 to 17 ° C. for about 1 week to prepare malt. Then, the malt is heated and dried to generate a unique color and aroma, and then crushed. After that, auxiliary materials such as rice and corn starch are added to saccharify the starch, hops are added to the filtered product, which is boiled and filtered to make wort. Next, the obtained wort is cooled,
About 1 to 10% by weight of the above-mentioned basidiomycetes is added, and fermentation and aging are performed under aerobic conditions (fermentation step). Fermentation conditions are usually 1
It is preferably at 5 to 25 ° C. for about 10 to 20 days. afterwards,
Similar to the conventional beer production method, a beer subjected to alcohol fermentation with the above basidiomycetes can be obtained through a filtration step, or after this step, a bottling or canning step and a sterilization step.

Further, taking wine (red wine) as an example,
Description will be given along the flow sheet shown in FIG. That is,
First, the grapes of dark red or black-purple varieties are crushed, and juice, peels and seeds are collected. Then, after adjusting the sugar content by supplementing sugar with sugar, glucose or the like, about 1 to 10% by weight of the above basidiomycete is added and fermented under aerobic conditions (fermentation step).
Fermentation conditions are usually preferably 20 to 25 ° C. for 7 to 10 days. Then, like the conventional wine production method, after pressing, post-fermentation, pulling, barrel storage, each step of filtration, by filling in a container such as bottles and cans, red wine alcohol-fermented by the basidiomycete can get.

The alcoholic beverage thus obtained is produced by alcoholic fermentation of the above-mentioned basidiomycete, and therefore β-D- which is a physiologically active substance derived from basidiomycete.
It contains glucan and the like. Therefore, when the above alcoholic beverage is drunk, it has a cancer prevention effect, a widespread epidemic prevention effect, an allergy prevention effect (effective for atopy),
The dietary fiber effect and the like can be obtained. Also, depending on the type of basidiomycete, it will contain a fibrinolytic enzyme that has the action of dissolving thrombus, and an antithrombin active substance that makes it difficult to form thrombus,
A preventive effect on thrombosis such as myocardial infarction and cerebral thrombosis can be obtained. Further, it becomes a substance containing ergosterol (provitamin D ₂ ) or exhibits antibacterial activity (improves preservability). And, when shiitake is used, it contains erytadenine, which has the effect of preventing hypertension and hypercholesterolemia, and when mannemtake is used, it contains polysaccharide ganodelan, which has a hypoglycemic effect. When it is present, it contains flamtoxin which has a cardiotonic action, and when it is used, it contains a nerve growth factor which has a dementia-improving effect. When maitake mushrooms are used, they exhibit a blood pressure-lowering action that reduces cholesterol levels, neutral fats, blood sugar levels, and urinary glucose levels, and when oyster mushrooms are used, they suppress food intake (lectin activity). Will be demonstrated.

The alcoholic beverage is produced using a specific basidiomycete, but has the same alcohol content as conventional alcoholic beverages. Moreover, a mushroom flavor that is not present in the flavor of conventional alcoholic beverages is imparted, and the olfactory sense is completely new. Furthermore, in terms of taste, by making the most of each ingredient,
It has a refreshing taste (beer), fruity taste (wine), and a rich taste (sake) with the subtle taste of basidiomycetes, making it a completely new product.

In the above production method, an example in which fermentation was carried out under aerobic conditions was explained, but the production method of the alcoholic beverage of the present invention is not limited to this, and fermentation can be carried out under anaerobic conditions as in the conventional method. It is also possible to do so. Under anaerobic conditions, the yeast used in the fermentation process in the conventional alcoholic beverage manufacturing method can be simply replaced with the specific basidiomycete, so that conventional manufacturing equipment can be used as it is, and new equipment can be used. It is possible to manufacture alcoholic beverages without adding more.

Next, examples will be described together with comparative examples.

[0034]

Example 1 First, 30 g of rice (Koshihikari from Toyama) was thoroughly washed, immersed overnight, drained, dispensed into an Erlenmeyer flask, and sterilized by an autoclave (120 ° C., 20 minutes). Then, the mycelium of the basidiomycete Agarikustake (Brazilian (directly imported) Agarikustake, manufactured by Eidai Pharmaceutical Co., Ltd.) was rubbed with a gauze-filled funnel in a clean bench, and sterilized rice To about 5% by weight. After that, cover with a sterilized sun cap sheet,
Room temperature (20
C.) and left for 15 days. Then, add 120 ml of sterilized water, cover with a sterilized sun cap sheet, and remove it at room temperature (2
It was shake-cultured at 0 ° C. for 29 days (aerobic condition). Then, the intended alcoholic beverage (sake) was produced by filtering the culture solution. Alcohol content is 8.0
%Met.

[0035]

Example 2 A target alcoholic beverage (sake) was produced in the same manner as in Example 1 except that the culture condition was static culture (anaerobic condition). The alcohol content was 8.0%.

[0036]

Example 3 An alcoholic beverage (sake) was produced in the same manner as in Example 1 except that Buna-shimeji mushroom (Super, Mabikoshimeji, JA Nagano Keiren) was used as the basidiomycete.

[0037]

[Example 4] An alcoholic beverage (sake) was produced in the same manner as in Example 1 except that Buna-shimeji mushroom (Super Yamabikoshimeji, JA Nagano Keizai Ren) was used as a basidiomycete and alcohol fermentation was performed under anaerobic conditions.

[0038]

[Example 5] 16 g of beer base [tasty beer base B (component: malt extract, hop), manufactured by NB Japan] was dissolved in 150 ml of water to obtain a sugar content of 7.6%,
A wort having a pH of 5.8 to 6.0 was prepared, dispensed in an Erlenmeyer flask, and sterilized by an autoclave (120 ° C., 20 minutes). Next, prepare basidiomycete Matsutake (collected at Tanba Sasayama), strain this mycelium with a funnel filled with gauze in a clean bench, and add about 5% by weight to the sterilized wort.
Inoculated. Then, the lid was covered with a sterilized suncap sheet, and shake culture was carried out at room temperature (20 ° C.) for 14 days (aerobic condition). Then, the intended alcoholic beverage (beer) was produced by filtering the culture solution. The alcohol content was 4.6%.

[0039]

Example 6 A target alcoholic beverage (sake) was produced in the same manner as in Example 5, except that the culture condition was static culture (anaerobic condition). The alcohol content was 4.6%.

[0040]

[Example 7] An alcoholic beverage (beer) was produced in the same manner as in Example 5 except that Tsukuritake (Mushroom, Mitsukura Agriculture and Forestry) was used as the basidiomycete.

[0041]

[Example 8] An alcoholic beverage (beer) was produced in the same manner as in Example 5, except that Tsukuritake (Mushroom, Mitsukura Norin) was used as the basidiomycete and alcohol fermentation was performed under anaerobic conditions.

[0042]

[Example 9] First, the grape (Kyoho) was removed from the tuft, washed well, crushed with a mixer, and gauze was squeezed into the funnel to obtain 30 ml of fruit juice. Then, adjust the sugar content of the juice to 11
Adjusted to%. Next, after adjusting the pH to 5.8 to 6.0, the mixture was dispensed into an Erlenmeyer flask and the autoclave (120
Sterilized at 20 ° C for 20 minutes. Next, prepare basidiomycetes, oyster mushrooms (sold from the Hyogo Plant Production Center),
The mycelium was rubbed with a funnel filled with gauze in a clean bench, and about 5% by weight of the sterilized fruit juice was inoculated. After that, cover with a sterilized suncap sheet and store at room temperature (20
The cells were shake-cultured for 10 days (aerobic condition). And
The intended alcoholic beverage (red wine) was produced by filtering the culture solution. The alcohol content is 12.
It was 2%.

[0043]

Example 10 The intended alcoholic beverage (red wine) was produced in the same manner as in Example 9 except that the culture condition was static culture (anaerobic condition). Alcohol content is 1
It was 2.2%.

[0044]

Example 11 An alcoholic beverage (red wine) was produced in the same manner as in Example 9 except that Ganoderma lucidum (sold by the Plant Production Center of Hyogo Prefecture) was used as the basidiomycete.

[0045]

[Example 12] An alcoholic beverage (red wine) was produced in the same manner as in Example 9 except that Ganoderma lucidum (sold by the Plant Production Center of Hyogo Prefecture) was used as a basidiomycete and alcohol fermentation was performed under anaerobic conditions.

[0046]

COMPARATIVE EXAMPLE 1 As Comparative Example 1, commercially available sake (manufactured by King Brewing Co., Noho, alcohol content 13.5%) was prepared.

[0047]

[Comparative Example 2] As Comparative Example 2, a commercially available beer (manufactured by Asahi Breweries, Iccho, alcohol content 4.5%) was prepared.

[0048]

Comparative Example 3 As Comparative Example 3, a commercially available wine (Santa Ana Cabernet Sauvignon, manufactured by Santa Ana Co., 12.5% alcohol) was prepared.

With respect to the alcoholic beverages of Examples and Comparative Examples thus obtained, as shown below,
Amylase activity, alcohol dehydrogenase activity, β-D-
The presence or absence of glucan, fibrinolytic activity, and antithrombin activity were measured and evaluated. The results are shown in Tables 1 to 4 below.

[Amylase activity] First, 5 mM C in which 2.5% by weight of soluble starch was dissolved
A 50 mM acetate buffer containing aCl ₂ was prepared and used as a substrate solution. Next, 100 μl of the crude enzyme solution was added to 300 μl of the substrate solution and reacted at 40 ° C. for 5 minutes. The enzymatic reaction was stopped by heating in boiling water for 10 minutes. After the reaction was stopped, 1.0 ml of 0.5 N acetic acid and 3.0 ml of 0.1% iodine solution were added to the reaction solution, and the mixture was stirred well and then the absorbance at 700 nm was measured with a double beam spectrophotometer. The value of Abs (700 nm) is the initial value (blank,
0.400) (value measured using water in place of the crude enzyme solution)
It was evaluated as having an amylase activity when it decreased from 0, and evaluated as having no amylase activity when not decreasing from 0.400. For example, the crude enzyme solution of Agaricus mushroom (Example 1) decreased to 0.049,
Since the crude enzyme solution of Matsutake (Example 5) decreased to 0.148, it could be confirmed that each had amylase activity.

[Alcohol dehydrogenase activity] * 1: Presence or absence of alcohol dehydrogenase First, basidiomycetes are subjected to ultrasonic treatment and centrifugal separation treatment,
The supernatant was prepared as a crude enzyme solution. Further, 12.5 ml of 0.1 M Tris buffer (pH 7.4) and NAD ⁺ 0.
1 ml, 0.1 M ethanol 11.0 ml, phenazine methosulfate (PMS) 0.5 ml, nitroblue tetrazolium (NBT) 0.5 ml, and water 10
A reaction solution was prepared by blending and mixing with ml. Next, after performing polyacrylamide gel electrophoresis using 0.02 ml of the crude enzyme solution, the polyacrylamide gel was immersed in the reaction solution and reacted at 30 ° C. for 1 hour. Then, the presence or absence of alcohol dehydrogenase activity was evaluated from the stained active band. That is, the one in which a stained alcohol dehydrogenase band was confirmed was marked as having alcohol dehydrogenase activity, and the one in which no band was confirmed was marked as having no alcohol dehydrogenase activity. * 2: Measurement of alcohol dehydrogenase activity First, a crude enzyme solution was prepared in the same manner as above. Also,
0.16 ml of 0.25M Tris buffer (pH 7.4)
, NAD ⁺ 0.05 ml, 0.1 M ethanol 0.
A reaction solution was prepared by mixing 1 ml and 0.19 ml of water and mixing them. Next, the reaction solution is heated to 30 ° C.
The reaction mixture was heated to 0.degree. C., and 0.05 ml of the crude enzyme solution which had been heated to 30.degree. And 340
The alcohol dehydrogenase activity (units / mg) was determined by measuring the change with time in absorbance at nm and determining the rate of increase in absorbance. In addition, 1 μm per minute
The amount of ole NAD ⁺ reducing enzyme was set to 1 unit.

[Presence or absence of β-D-glucan] In order to confirm whether or not β-D-glucan is present in an alcoholic beverage, high performance liquid chromatography (H
Β-D-glucan was detected using PLC). Then, the one in which the peak of β-D-glucan could be confirmed was ◯, the one in which it could not be confirmed or some peaks were confirmed in other experiments [enzyme (β-1,3-glucanase)
Treatment experiments] confirmed that the peak was not β-D-glucan, and were marked with x.

[Fibrinolytic activity] 0.03 ml of an alcoholic beverage was sprinkled in a circular shape on the surface of a fibrin plate (5671 mm ² ) (spraying area 19.6 m 2
m ² ) The dissolution area after standing for 1 hour was measured.

[Antithrombin activity] The time (thrombin time) until fibrinogen was changed to fibrin by the reaction of thrombin and coagulation was measured. That is, 0.13% by weight fibrinogen solution 0.4
0.1 ml of alcoholic beverage was allowed to act on ml, and the time until coagulation was measured.

[0055]

[Table 1]

[0056]

[Table 2]

[0057]

[Table 3]

[0058]

[Table 4]

From the results shown in Tables 1 to 4, alcoholic beverages (Examples 1 to 12) produced by alcoholic fermentation with basidiomycetes having amylase activity and alcohol dehydrogenase activity are physiologically active substances. It can be seen that β-D-glucan is contained. On the other hand, commercially available sake, beer, and wine (Comparative Examples 1 to 3) have β-
It can be seen that D-glucan is not included. It can also be confirmed that the alcoholic beverages of Examples 1 to 12 exhibit fibrinolytic activity. Further, it can be confirmed that the alcoholic beverages of Examples 1 to 12 have antithrombin activity as compared with commercially available sake, beer, and wine (Comparative Examples 1 to 3).

[0060]

As described above, the method for producing an alcoholic beverage of the present invention uses a basidiomycete having an amylase activity and an alcohol dehydrogenase activity in place of the yeast which has been conventionally used in the alcohol fermentation step, Alcohol fermentation is performed by this basidiomycete. for that reason,
It is possible to obtain a completely new alcoholic beverage containing an anti-cancer substance (β-D-glucan) and the like that cannot be produced by yeast. Therefore, it is possible to provide a functional and healthy beverage that has never existed before.

The use of basidiomycetes has the advantage that it is possible to produce alcoholic beverages not only under anaerobic conditions but also under aerobic conditions. Therefore, the manufacturing conditions of alcoholic beverages can be eased. If alcohol fermentation is performed with specific basidiomycetes under anaerobic conditions,
There is an advantage that conventional manufacturing equipment can be used as it is, and alcoholic beverages can be manufactured without adding new equipment.

[Brief description of drawings]

FIG. 1 is a flow sheet showing a method for producing an alcoholic beverage of the present invention.

FIG. 2 is a flow sheet showing a procedure of a method for producing an alcoholic beverage (sake) according to the present invention.

FIG. 3 is a flow sheet showing a procedure of a method for producing an alcoholic beverage (beer) of the present invention.

FIG. 4 is a flow sheet showing a procedure of a method for producing an alcoholic beverage (wine) of the present invention.

FIG. 5 is a flow sheet showing a conventional procedure for producing sake.

FIG. 6 is a flow sheet showing a procedure of a conventional beer production method.

FIG. 7 is a flow sheet showing a procedure of a conventional wine manufacturing method.

Claims (5)

(57) [Claims] 1. A method for producing an alcoholic beverage in which alcoholic fermentation is performed by a bacterium having an alcoholic fermentation ability, wherein the bacterium having an alcoholic fermentation ability is a basidiomycete having an amylase activity and an alcohol dehydrogenase activity. Alcoholic beverage manufacturing method. 2. The amylase activity and alcohol desorption
Basidiomycetes with hydrogen enzyme activity
Bamboo, enokitake, matsutake, jellyfish, chestnut, shii
Bamboo shoots, gyoza, suehirotake, taro mushrooms, butterflies
Rei Maitake, Bukyou, Beech Shimeji, Maitake, Ma
Bamboo shoots, Ganoderma lucidum, Tsukuritake mushrooms, Salix matsutake mushrooms,
Yamabushitake, Kawaratake, Kobukisarunokeshi, Na
A few selected from the group consisting of Meko and Numerisugitake
The method for producing an alcoholic beverage according to claim 1, which is at least one kind . 3. A method according to claim 1 or 2, wherein the alcoholic beverage performing alcohol fermentation under aerobic conditions. 4. A method of performing alcoholic fermentation under anaerobic conditions.
Item 1. A method for producing an alcoholic beverage according to item 1 or 2. 5. The method according to any one of claims 1 to 4.
An alcoholic beverage obtained by the method for producing a rucor beverage.