JP3362312B2 - Beer production method and beer obtained thereby - Google Patents

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for producing beer using a basidiomycete having an alcohol dehydrogenase activity and a beer obtained thereby.

[0002]

2. Description of the Related Art Conventionally, yeast is used in the production of beer for the purpose of carrying out alcohol fermentation. In general, beer is obtained by germinating barley to make malt, saccharifying barley starch, and then alcohol-fermenting wort containing hops. For example, it is manufactured along the flow sheet shown in FIG. That is, first, barley is soaked in water to absorb water, and then germinated at 13 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, heat the malt,
Grind after drying to produce a unique color and scent. After that, saccharified starch is added by adding auxiliary materials such as rice and corn starch, and hops are added to the filtered product and boiled.
Filter to make wort. Then, the wort is cooled and yeast is added to ferment it. 15 in case of fermentation by top yeast
In the case of fermentation with bottom yeast at ~ 20 ° C for 4-6 days,
Aging is performed 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.

[0003]

As described above, beer is conventionally produced using yeast having an alcohol-fermenting ability, but there are almost no reports of producing it using a bacterium other than yeast, and in particular basidiomycetes. It is not an exaggeration to say that there is no report. Moreover, since yeast has a small alcohol fermentation ability under aerobic conditions, there are restrictions on beer production conditions. Furthermore, β-D-glucan, which is an anti-cancer substance, is hardly present in beer produced using yeast, and further, a preventive effect on thrombosis such as myocardial infarction and cerebral thrombosis cannot be expected.

The present invention has been made in view of the above circumstances, and provides a completely new beer production method in which basidiomycetes are used in place of yeast in the fermentation step in the production of beer and beer obtained thereby. Is its purpose.

[0005]

In order to achieve the above-mentioned object, the present invention provides a first method for producing beer in which alcohol fermentation is performed by a basidiomycete having alcohol dehydrogenase activity in a fermentation step in the production of beer. Use as a summary.

A second aspect of the present invention is a beer obtained by the above-mentioned beer production method.

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, one having alcohol dehydrogenase activity exists. Then, as a result of trying to produce beer using such basidiomycetes, it was found that beer having the same alcohol content as the conventional one was obtained, and the present invention was reached.

Using the above-mentioned basidiomycete, beer can be produced under anaerobic conditions as in the conventional beer production.
Further, beer can be produced even under aerobic conditions. Therefore, there is an advantage that the beer production conditions 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 beer can be manufactured without adding new equipment.

Since the beer of the present invention is produced by a specific basidiomycete, not by the conventionally used yeast, it contains useful physiologically active substances which cannot be produced by yeast. That is, β- which exhibits antitumor properties, immune activity, antiallergic properties, and dietary fiber effects
D-glucan, chitin, heteropolysaccharide [pectic, hemicellulose, active hemicellulose complex (AHCC, Active hemicellulose compl
ex), polyuronide, lignin] and the like. Therefore, the beer of the present invention becomes a novel product which has never been available 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.

[0012]

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

The beer production method of the present invention uses a specific basidiomycete in the fermentation step in the conventionally known various beer production methods, and does not use a yeast that has been generally used in the past. Alcohol fermentation is performed.

The specific basidiomycete used in the present invention is not particularly limited as long as it has alcohol dehydrogenase activity, and for example, Agaricus blazei collected in forests in various places. ,
Pleurotus ostreatus (Pleurotus ostreatus), Enokitake (Flammulina velutipes), Tricholoma matsutak (Tricholoma matsutak)
e, Torikoromatsu Matsutake, Asteraceae (Auricularia a)
uricula, aricularia aricula), Kuritake (N
aematoloma sublateritium, Naematoloma subtilisum, Shiitake mushroom (Lentinus edodes, Lentinas eddes), Shouro (Rhizopogon rubescens, Rhizopogon rubescens), Suehirotake (Schizophyllum)
commune, schizophyllum commune, Pleu
rotus citrinopileatus, Plerotos citrinopileatus), butterflies (Dendropolyporus umbellat)
us, Dendro Polyporus Uberatas), Bukyou (Poria cocos, Polyacokos), Beech Shimeji (Hyps)
izygus marmoreus, Hypsizygus marmores, Maitake (Grifola frandosa),
Mustache (Laetiporus sulphureus), Ganoderma lucidum (Ganoderma lucidum), Tsukuritake (Agaricus bisporus)
, Agaricus bisporus), Salix matsutake (Agroc)
Ybe cylindracea, Aguroshibe cylindracea), Ya <br/> Ma Bed Shitake (Hericium erinaceum, Herishiumu Erinasemu), Coriolus versicolor (Coriolus versicolor, Koriorusu Vuyajikaraa), Kopf key Polyporaceae (Elfvingi
a applanata, Elfingia aplanata), nameko (Pholiota nameco, polio nameko), numerisugitake (Pholiota adiposa, poliota adiposa), aageki jellyfish (Auricularia polytricha, alicularia politrisha), white shimeji (Tricholoma japonic)
um, Tricoloroma japonicaum), Lactariu (Lactariu)
s volemus, Lactarius boremus), Numeiriguchi (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, Ganoderma neo japonicum), Hanabirattake (Sparassis crispa, Sparassis)
Crispa), Pleurotus cornucopia (Cordyceps militaris), and the like. Above all,
In consideration of availability, alcohol dehydrogenase activity, and the like, enoki mushroom, matsutake mushroom, mustard mushroom, and tsukuri mushroom are preferable. The use of Tsukuritake not only produces beer with a refreshing taste and aroma, but also has an advantage that various physiologically active substances are contained in beer.

Alcohol dehydration in the above specific basidiomycetes
The presence or absence of elementary enzyme activity can be confirmed, for example, as follows.
You can That is, first, ultrasonic waves are applied to basidiomycetes.
Process and centrifuge to prepare the supernatant as crude enzyme solution.
To make. In addition, 0.1 M Tris buffer (pH 7.4) 1
2.5 ml, NAD ⁺ 0.1 ml, 0.1 M ethanol
11.0 ml and phenazine methosulfate (P
0.5 ml of MS) and nitroblue tetrazolium (N
BT) 0.5 ml and water 10 ml are mixed and mixed.
To prepare a reaction solution. Next, the crude enzyme
Polyacrylamide gel electrophoresis using 0.02 ml of liquid
And then run the polyacrylamide gel on the above reaction.
Immerse in the solution and react at 30 ° C. for 1 hour. And dye
If it can be confirmed band of has been A alcohol dehydrogenase,
It can be said that it has alcohol dehydrogenase activity.
Even if the basidiomycetes listed above are used,
In the present invention, dehydrogenase activity is not confirmed.
It cannot be used.

Among the above basidiomycetes, those having an alcohol dehydrogenase activity of 1.0 units / mg or more are preferable, and those having 10.0 units / mg or more are particularly 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 beer. The alcohol dehydrogenase activity was determined by using a supernatant obtained by subjecting basidiomycetes to ultrasonic treatment and centrifugation as a crude enzyme solution, using ethanol, Tris buffer, NAD ^+, etc., and measuring 340 nm of NADH. It is determined by measuring the absorbance at.

The beer production method of the present invention can be carried out using the above-mentioned basidiomycetes, for example, as follows. That is, first, as shown in FIG. 1, 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 basidiomycete is added, and fermentation and aging are performed under anaerobic conditions (fermentation step). As the basidiomycete, it is preferable to use a pure culture that is liquid-cultured and grown as a mycelium. Fermentation conditions are usually 15 to 25 ° C and 1
About 0 to 20 days is preferable. Thereafter, as in the conventional beer production method, a beer subjected to alcohol fermentation with the above basidiomycetes is obtained through a filtration step, or after this step, a bottling or canning step and a sterilization step.

Since the beer thus obtained is produced by alcoholic fermentation of the above basidiomycete, it contains β-D-glucan which is a physiologically active substance derived from basidiomycete. For this reason, drinking the above-mentioned beer provides a cancer prevention effect, a widespread disease prevention effect, an allergy prevention effect (effective for atopy), a dietary fiber effect, and the like. Further, depending on the type of basidiomycete, a fibrinolytic enzyme having an action of dissolving a thrombus and an antithrombin active substance that makes it difficult to form a thrombus are contained, and a thrombosis preventive effect such as myocardial infarction or cerebral thrombosis can be obtained. In addition, it contains ergosterol (provitamin D ₂ ),
It exhibits antibacterial activity (improves shelf life). 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.

Further, the above-mentioned beer is produced using a specific basidiomycete, but has the same alcohol content as conventional beer. Moreover, a mushroom flavor that is not in the conventional beer flavor is added,
It will be completely new olfactively. Furthermore, in terms of taste, the taste is refreshing, and the subtle taste of each basidiomycete is added, resulting in a completely new taste.

Further, according to the above-mentioned manufacturing method, since the yeast used in the fermentation step in the conventional beer manufacturing method is simply replaced with the above-mentioned specific basidiomycete, the conventional manufacturing equipment can be used as it is, Beer can be manufactured without adding new equipment.

In the above production method, an example in which the fermentation step was carried out under anaerobic conditions was explained, but the production method of beer of the present invention is not limited to this, and fermentation can also be carried out under aerobic conditions. Is. This is because the specific basidiomycete can perform alcoholic fermentation even under aerobic conditions.

Next, examples will be described together with comparative examples.

[0023]

Example 1 According to the flow sheet shown in FIG. 1, beer was produced as follows. That is, first, malt prepared from barley was prepared. Then, the sugar content of malt was adjusted to 7.6% by weight, and the pH was further adjusted to 5.8 to 6.0.
After adjusting to a certain degree, heat treatment was performed to prepare wort. Then, about 5% by weight (to wort) of Matsutake mushroom (collected at Tanba Sasayama), which is a basidiomycete, was added to the obtained wort at room temperature (2
Alcoholic fermentation was performed under anaerobic conditions at 0 ° C. for 14 days. In this way, the intended beer was produced. The alcohol content was 4.6%.

[0024]

Example 2 Beer was produced in the same manner as in Example 1 except that the alcohol fermentation was carried out under aerobic conditions. The alcohol content was 4.6%.

[0025]

Example 3 Beer was produced in the same manner as in Example 1 except that Mustake (collected in Ashiodani, Shiga Prefecture) was used as the basidiomycete.

[0026]

[Example 4] Beer was produced in the same manner as in Example 1 except that Mustake (collected in Ashiodani, Shiga Prefecture) was used as a basidiomycete and alcohol fermentation was performed under aerobic conditions.

[0027]

Example 5 Beer was produced in the same manner as in Example 1 except that Enokitake mushrooms (sold by the Japan Mushroom Center) were used as basidiomycetes.

[0028]

Example 6 Beer was produced in the same manner as in Example 1 except that Enokitake mushrooms (sold by the Japan Mushroom Center) were used as basidiomycetes and alcohol fermentation was carried out under aerobic conditions.

[0029]

Example 7 Tsukuritake (Mushroom, Mitsuura Norin) was used as a basidiomycete and beer was produced in the same manner as in Example 1.

[0030]

[Example 8] Beer was produced in the same manner as in Example 1 except that Tsukuritake (Mushroom, Mitsuura Norin) was used as a basidiomycete and alcohol fermentation was performed under aerobic conditions.

[0031]

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

With respect to the thus-obtained beers of Examples and Comparative Examples, measurement and evaluation of alcohol dehydrogenase activity, presence / absence of β-D-glucan, fibrinolytic activity and antithrombin activity were carried out as follows. I went. The results are shown in Tables 1 and 2 below.

[Alcohol dehydrogenase activity] * 1: Presence or absence of alcohol dehydrogenase First, basidiomycetes are subjected to ultrasonic treatment and centrifugation 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 beer, high performance liquid chromatography (HPLC) is used.
Was used to detect β-D-glucan. And β
A peak was observed for -D-glucan, and it was confirmed that the peak was present in the beer. Note that FIG. 2 is data of beer produced using Matsutake (Example 1 product), and it can be confirmed that a β-D-glucan peak is present in a portion indicated by an arrow. Further, FIG. 3 is data of commercially available beer (comparative example), and some peaks are recognized in the portions indicated by arrows.
However, it has been confirmed that the peak shown by the data of commercially available beer is not β-D-glucan as a result of the enzyme (β-1,3-glucanase) treatment.

[Fibrinolytic activity] Beer was added on the surface of a fibrin flat plate (5671 mm ² ) in an amount of 0.
Sprayed with 03ml circular (scatter area 19.6 mm ^2),
The dissolution area after standing for 1 hour was measured.

[Anti-thrombin activity] The time (thrombin time) until fibrinogen was changed from fibrinogen 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 beer was allowed to act on ml and the time until coagulation was measured.

[0037]

[Table 1]

[0038]

[Table 2]

From the results of Tables 1 and 2, the beer (Examples 1 to 8) produced by alcoholic fermentation with basidiomycetes having alcohol dehydrogenase activity has β-D- which is a physiologically active substance. You can see that it contains glucan. In contrast, commercially available beer (comparative example product) has β
It can be seen that -D-glucan is not included. Also,
It can also be confirmed that the beer (Examples 4, 7, and 8) produced by using master bamboo and Tsukuritake as basidiomycetes show fibrinolytic activity. Furthermore, beer produced using Matsutake mushrooms, mustache mushrooms, enokitake mushrooms, and Tsukuritake mushrooms as basidiomycetes (Examples 2, 4, 5 to 8 products) have antithrombin activity as compared with commercially available beers (comparative example products). You can also confirm that it is shown.

Next, the antibacterial activity test was carried out on the above-mentioned products of Examples 3, 4 and Comparative Example as follows. The results are shown in Table 3 below.

[Antibacterial Activity Test] Fungal fungus, Black mold (Aspergillus niger), fungal yeast (Saccharomyces)
cerevisiae, Saccharomyces cerevisiae, Gram-negative bacteria Escherichia coli, and Gram-positive bacteria Bacillus natto.
The antibacterial activity against natto and Bacillus natto was evaluated. Specifically, first, the standard agar medium on which each of the above-mentioned bacteria can grow was kept warm at 55 ° C., then each of the bacteria was transplanted, poured into a Petri dish with a penicillin cup set up, and solidified. After solidification, remove the penicillin cup and into the hole,
Beer was added and each fungus was cultured under standard conditions. As a result, ◯ was given to those in which a blocking circle was formed (which showed antibacterial activity), and x was given to those in which no blocking circle was formed (which did not show antibacterial activity).

[0042]

[Table 3]

From the results shown in Table 3, it can be seen that the beers produced using master salmon as basidiomycetes (Examples 3 and 4) have antibacterial activity against black mold, yeast and E. coli. On the other hand, it can be seen that the comparative example product has no antibacterial activity against black mold, yeast, Escherichia coli, and Bacillus natto.

[0044]

INDUSTRIAL APPLICABILITY As described above, in the production of beer of the present invention, in the fermentation step in the conventional production of beer, alcohol fermentation is not performed by yeast but by basidiomycetes having alcohol dehydrogenase activity. Is to do. Therefore, a completely new beer containing an anti-cancer substance (β-D-glucan) and the like that cannot be produced by yeast can be obtained. Therefore, it is possible to provide a functional and healthy beverage that has never existed before.

The use of basidiomycetes has the advantage that beer can be produced not only under anaerobic conditions but also under aerobic conditions. Therefore, beer production conditions can be relaxed. Further, if alcohol fermentation is carried out with a specific basidiomycete under anaerobic conditions, there is an advantage that conventional manufacturing equipment can be used as it is and beer can be manufactured without adding new equipment.

[Brief description of drawings]

FIG. 1 is a flow sheet showing a procedure of a method for producing beer according to the present invention.

FIG. 2 is a chart of beer as an example product.

FIG. 3 is a chart of beer as a comparative example product.

FIG. 4 is a flow sheet showing a procedure of a conventional beer production method.

Claims (5)

(57) [Claims] 1. A method for producing beer, characterized in that, in a fermentation step in the production of beer, alcohol fermentation is carried out by a basidiomycete having alcohol dehydrogenase activity. 2. Having the alcohol dehydrogenase activity
Basidiomycetes, Agaricus tussalis, oyster mushroom, enoki mushroom, ma
Ivy, mushroom, chrysanthemum, shiitake, ginger, su
Ehirotake, Pleurotus cornucopiae, Choreimaitake, Bukuri
Leopard, beech, maitake, mustache, mannenta
Bamboo, Tsukuritake, Salix matsutake, Yamabushitake, Kawa
Ratake, Coffis sarnoche, sardine, numeris-gita
Moss, moss jellyfish, shimejimeji, shiitake mushroom, numeri
Guchi, matsutake mushroom, oyster mushroom, pine tree, mukitsuke, ya
Brebenitatake, liquorice, octopus, Hanabira
At least one selected from the group consisting of bamboo and pupa
The method for producing beer according to claim 1, wherein the beer is also a kind. 3. A beer method according to claim 1 or 2, wherein performing the alcohol fermentation in anaerobic conditions. 4. A method of performing alcoholic fermentation under aerobic conditions.
Item 1. A method for producing beer according to Item 1 or 2. 5. The bis according to any one of claims 1 to 4.
Beer obtained by the brewing method.