JP2001286274A - Method for producing wine and wine obtained by the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an entirely new method for producing wine using basidiomycete instead of yeast. SOLUTION: This method comprises conducting alcoholic fermentation using basidiomycete having alcohol dehydrogenase activity in a wine-producing fermentation process.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a method for producing wine using basidiomycetes having alcohol dehydrogenase activity, and to a wine obtained thereby.

[0002]

2. Description of the Related Art Conventionally, yeast has been used in wine production for the purpose of performing alcohol fermentation. Wine is generally obtained by fermenting grape juice using yeast, and is roughly classified into red wine and white wine.

[0003] Red wine is produced, for example, according to a flow sheet shown in FIG. That is, first, the grape of dark red or black purple variety is crushed, and juice, pericarp, and seeds are collected. Then, a sulfite compound (potassium metabisulfite or the like) is added for the purpose of preventing the growth and oxidation of various bacteria, and sugar is adjusted with sugar, glucose or the like, and then the sugar content is adjusted. Ferment at 7 ° C for 7-10 days. After that, by pressing, post-fermentation, origination, barrel storage, and filtration, by filling in containers such as bottles,
Red wine is obtained.

[0004] White wine is produced in substantially the same manner as red wine, but is obtained by fermenting only fruit juice with yeast from green-yellow varieties of grapes, and is usually fermented at 15 ° C for about 3 weeks. It is obtained by squeezing, squeezing, sagging and aging.

[0005]

As described above, wine has been conventionally produced using yeast having an alcohol fermentation ability. However, there has been almost no report that wine has been produced using a fungus other than yeast. It is no exaggeration to say that there is no report at all. In addition, yeast has a low alcohol fermentation ability under aerobic conditions, so that the conditions for producing wine are limited. Furthermore, wine produced using yeast has almost no β-D-glucan, which is an anticancer substance, and cannot be expected to prevent thrombosis such as myocardial infarction and cerebral thrombosis.

[0006] The present invention has been made in view of such circumstances, and in the alcohol fermentation step, instead of yeast,
An object of the present invention is to provide a completely new method for producing wine using basidiomycetes and to provide a wine obtained thereby.

[0007]

In order to achieve the above-mentioned object, the present invention relates to a first method for producing wine in which alcohol fermentation is carried out by a basidiomycete having alcohol dehydrogenase activity in a fermentation step in wine production. Make a summary.

[0008] A second aspect of the present invention is a wine obtained by the above-described wine production method.

The present inventors have repeated various experimental studies on basidiomycetes producing useful physiologically active substances. In the process, basidiomycetes were conventionally thought to have no alcohol fermentation ability. However, among various basidiomycetes, it was found that those having alcohol dehydrogenase activity existed. Then, as a result of attempting to produce wine using such basidiomycetes, the present inventors have found that wine having an alcohol content comparable to that of the related art can be obtained, and have reached the present invention.

When the basidiomycete is used, wine can be produced under anaerobic conditions as in the conventional wine production.
In addition, wine can be produced even under aerobic conditions. Therefore, there is an advantage that the conditions for producing wine can be eased. In the case of anaerobic conditions, there is an advantage that the conventional manufacturing equipment can be used as it is and wine can be manufactured without adding new equipment.

[0011] The wine of the present invention is produced by a specific basidiomycete rather than a conventionally used yeast, and therefore contains a useful physiologically active substance that cannot be produced by yeast. That is, β- which exerts antitumor, immunological, antiallergic and dietary fiber effects
D-glucan, chitin and heteropolysaccharide exhibiting a dietary fiber effect [pectic, hemicellulose, active hemicellulose complex (AHCC, Active hemicellulose compl)
ex), polyuronide, lignin]. Therefore, the wine of the present invention becomes a completely new one that has never been seen before, and can be a functional and healthy beverage.

In the present invention, basidiomycetes refer to those whose fruiting body is called "mushroom", and include basidiomycetes (haratake mushrooms, Hidanashi mushrooms, abdomen fungi, rhododendrons) in addition to basidiomycetes in microbial taxonomy. It is used in the concept including a part of ascomycetes.

In the present invention, the anaerobic condition is defined as
It refers to a condition where the oxygen partial pressure is less than 1.0 kPa, and includes a state where oxygen is completely absent. On the other hand, aerobic conditions refer to conditions where the oxygen partial pressure is 1.0 kPa or more.

[0014]

Next, an embodiment of the present invention will be described.

The method for producing wine of the present invention uses a specific basidiomycete in a fermentation step in various conventionally known wine production methods, instead of using a yeast generally used in the past, and using this basidiomycete. It performs alcohol fermentation.

The specific basidiomycete used in the present invention is not particularly limited as long as it has an alcohol dehydrogenase activity. For example, Agaricus blazei (Agaricus blazei, Agaricus blazei) collected from forests and the like in various places. ,
Pleurotus ostreatus (Pleurotus ostreatus), Enokitake (Flammulina velutipes), Matsutake (Tricholoma matsutak)
e, Tricholoma matsutake, jellyfish (Auricularia a
uricula, Aricularia alicula, Kuritake (N
aematoloma sublateritium, Naematoma subrateritium, Shiitake mushroom (Lentinus edodes, Lentinas eddes), Shouro (Rhizopogon rubescens, Rhizopogon rubesensu), Suehirotake mushroom (Schizophyllum)
commune, Shizofiram komune), Tamagitake (Pleu)
rotus citrinopileatus, Prelotos citrinopiretas, Butterfly Maitake (Dendropolyporus umbellat)
us, dendropoliporus uberatasu, porcelain (Poria cocos, polia cocos), bunashimeji (Hyps)
izygus marmoreus, Hypsizigas marmores), Maitake (Grifola frandosa, Grifola frandosa),
Mustard (Laetiporus sulphureus, Laetiporas sulphreus), Ganoderma lucidum (Ganoderma lucidum), Amanita mushroom (Agaricus bisporus)
, Agaricus bisporus), Willow matsutake (Agroc)
ybe cylindracea, Agrosibe cylindracea, Yamapushitake (Hericium erinaceum), Kawaratake (Coriolus versicolor)
a applanata, Elfingia apranatha, nameko (Pholiota nameco, poliotanameko), squirrel mushroom (Pholiota adiposa, poliota adiposa), Amitake (Suillus bovinus, Suils bobinas), Shiroki jellyfish (Tremella fuciformis, Plesio sulcus, Tremella fuciformis) Impdekas, Nishikiaritake (Pseudohydnum)
gelatinosum, pseudohydramum geratinosam, Novoliru (Helvella crispa, Helvera crispa), Bunaharitatake (Mycoleptodonoides aitchisonii)
, Mycoleptdonoides Aichisonii), Hon Shimeji (Lyophyllumshimeji),
Matsuba Haritake (Bankera fuligineo-alba, Bankera)
Figineo alba), Molinokarebatake (Collybia d)
ryophila, Kolivia doriophylla, Polyporus tuberaster, Rozites caperatus, Logites
Capellatas, Pleurocybella porrigen
s, Pleurosibera polygens)
hyllum fumosum). Among them, in consideration of availability, alcohol dehydrogenase activity, and the like, agarictake, enokitake, oyster mushroom, and mannaketake are preferred. The use of Ganoderma radiata not only produces wine with a fruity taste and aroma, but also has the advantage that the wine contains a bioactive substance that is effective as a crude drug.

Among the above basidiomycetes, those having an alcohol dehydrogenase activity of 1.0 units / mg or more are preferable, and those having an alcohol dehydrogenase activity of 10.0 units / mg or more are 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 for wine production. The alcohol dehydrogenase activity was determined by using a supernatant obtained after basidiomycete sonication and centrifugation as a crude enzyme solution, using ethanol, Tris buffer, NAD ^{+ and the} like, and measuring 340 nm indicated by NADH. Is determined by measuring the absorbance at

The method for producing wine according to the present invention can be carried out using the basidiomycete described above, for example, as follows.

In the case of red wine, as shown in FIG. 1, first, grapes of dark red or black purple varieties are crushed, and fruit juice,
Collect pericarp and seeds. Then, after the sugar content is adjusted by sugar supplementation with sugar, glucose, etc., the basidiomycete is added in an amount of 1 to 10% by weight.
In addition, fermentation is performed under anaerobic conditions (fermentation step). It is preferable to use a basidiomycete obtained by subjecting pure culture to liquid culture and growing as mycelium. In addition, fermentation conditions are preferably preferably about 20 to 25 ° C. for about 7 to 10 days. Then, as with conventional wine production, squeezing, post-fermentation,
Red wine that has been subjected to alcohol fermentation by the basidiomycete is obtained by packing in containers such as bottles and cans through the steps of wiping, storage, and filtration.

In the case of white wine, as shown in FIG. 2, first, grapes of a green-yellow variety are crushed, and juice, peel,
Collect the seeds. Then, use a juicer to take only the juice. Next, after sugar supplementation is performed to adjust the sugar content, about 1 to 10% by weight of the basidiomycete is added and fermentation is performed under anaerobic conditions (fermentation step). Basidiomycetes are liquid cultures of pure cultures,
It is preferred to use those grown as mycelia.
The fermentation conditions are usually preferably at 13 to 18 ° C. for about 3 weeks. After that, as in the conventional wine manufacturing method, the white wine is subjected to alcohol fermentation by the above basidiomycete by passing through the steps of pulling, storing, and filtering, and packing the bottle in a container such as a bottle.

Since the wine thus obtained is produced by alcohol fermentation of the basidiomycete, it contains β-D-glucan, which is a physiologically active substance derived from basidiomycete. For this reason, drinking the above-mentioned wine provides a cancer prevention effect, a widespread plague prevention effect, an allergy prevention effect (effective for atopy), a dietary fiber effect, and the like. In addition, depending on the type of basidiomycetes, the basidiomycetes contain a fibrinolytic enzyme having an action of dissolving a thrombus and an antithrombin active substance which makes it difficult to form a thrombus. In addition, it may contain ergosterol (provitamin D ₂ ),
It will exhibit antibacterial activity (enhancing shelf life). When shiitake is used, it contains erytadenine, which has an effect of preventing hypertension and hypercholesterolemia, and when mannenthus is used, it contains polysaccharide ganoderan, which has a hypoglycemic effect. In this case, it contains a flamtoxin having a cardiotonic effect, and when Yamabushitake is used, it contains a nerve growth factor having an effect of improving dementia. In addition, when maitake is used, it exerts a blood pressure lowering effect of reducing cholesterol, triglyceride, blood sugar, and urine sugar levels, and when oyster mushroom is used, food intake inhibitory activity (lectin activity) Will be demonstrated.

The above-mentioned wine is produced using a specific basidiomycete, but has a similar alcohol content to conventional wine. Moreover, it has a mushroom flavor that is not in the flavor of conventional wine,
It will be completely new in olfaction. Furthermore, the taste becomes fruity, and the subtle taste of each basidiomycete is added, and it becomes completely novel.

Further, according to the above-mentioned production method, it is only necessary to replace the yeast used in the fermentation step in the conventional wine production method with the above-described specific basidiomycete, so that the conventional production equipment can be used as it is. Wine production can be performed without adding new equipment.

In the above-mentioned production method, an example in which fermentation is performed under anaerobic conditions has been described. However, the method for producing wine of the present invention is not limited to this, and fermentation can be performed under aerobic conditions. is there. This is because the basidiomycetes can perform alcohol fermentation even under aerobic conditions.

Next, examples will be described together with comparative examples.

[0026]

Example 1 Wine was produced in the following manner according to the flow sheet shown in FIG. That is, first, grapes (Kyoho) were crushed with a mixer to obtain fruit juice. Then
The sugar content of the juice was adjusted to 11% by weight, and the pH was further adjusted to 5.8.
After adjusting to about 6.0, a heat treatment was performed. The obtained juice is inoculated with basidiomycete oyster mushroom (available from the Hyogo Prefectural Plant Production Center) at about 5% by weight (based on the juice), and is anaerobically incubated at room temperature (20 ° C.) for 10 days. Alcohol fermentation was performed. Thus, the target wine (red wine) was produced. The alcohol content is 1
2.2%.

[0027]

Example 2 A wine was produced in the same manner as in Example 1 except that alcohol fermentation was performed under aerobic conditions. The alcohol content was 12.2%.

[0028]

Example 3 A wine was produced in the same manner as in Example 1 except that enokitake was used as a basidiomycete (available from Nippon Mushroom Center).

[0029]

Example 4 A wine was produced in the same manner as in Example 1, except that enokitake was used as a basidiomycete (available from Nippon Mushroom Center) and alcohol fermentation was performed under aerobic conditions.

[0030]

[Example 5] Matsutake as basidiomycete (collected at Tanba Sasayama)
A wine was produced in the same manner as in Example 1, except for using.

[0031]

[Example 6] Matsutake as basidiomycete (collected at Tanba Sasayama)
And wine was produced in the same manner as in Example 1 except that alcohol fermentation was performed under aerobic conditions.

[0032]

Example 7 A wine was produced in the same manner as in Example 1 except that agaricus stamen (Agaricus stamen from Brazil (directly imported), manufactured by Eisai Pharmaceutical Co., Ltd.) was used as the basidiomycete.

[0033]

[Example 8] Agarixtake (Agaricutake from Brazil (directly imported), manufactured by Eisai Pharmaceutical Co., Ltd.) was used as a basidiomycete.
A wine was produced in the same manner as in Example 1 except that alcohol fermentation was performed under aerobic conditions.

[0034]

Example 9 A wine was produced in the same manner as in Example 1, except that mussels (collected from Ashioya, Shiga Prefecture) were used as basidiomycetes.

[0035]

Example 10 A wine was produced in the same manner as in Example 1, except that mash was used as a basidiomycete (collected from Ashioya, Shiga Prefecture) and alcohol fermentation was performed under aerobic conditions.

[0036]

Example 11 A wine was produced in the same manner as in Example 1 except that Mannentake was used as a basidiomycete (available from the Hyogo Prefectural Plant Production Center).

[0037]

Example 12 A wine was produced in the same manner as in Example 1 except that menthol mushrooms (available from the Hyogo Prefectural Plant Production Center) were used as basidiomycetes and alcohol fermentation was performed under aerobic conditions.

[0038]

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

The wines of Examples and Comparative Examples thus obtained were evaluated for alcohol dehydrogenase activity, presence or absence of β-D-glucan, fibrinolytic activity, and antithrombin activity, as described below. Was done. The results are shown in Tables 1 to 3 below.

[Alcohol dehydrogenase activity] * 1: Presence or absence of alcohol dehydrogenase First, basidiomycete was subjected to ultrasonic treatment and centrifugation.
The supernatant was prepared as a crude enzyme solution. Also, 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, nitro blue tetrazolium (NBT) 0.5 ml, water 10
Then, a reaction solution was prepared by mixing and mixing the resulting mixture with the same. Next, polyacrylamide gel electrophoresis was performed using 0.02 ml of the above crude enzyme solution, and then the polyacrylamide gel was immersed in the above 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, those in which a stained alcohol dehydrogenase band could be confirmed were evaluated as ○ as having alcohol dehydrogenase activity, and those in which no band could be confirmed were evaluated as x as having no alcohol dehydrogenase activity. * 2: Measurement of alcohol dehydrogenase activity First, a crude enzyme solution was prepared in the same manner as described above. Also,
0.16 ml of 0.25 M Tris buffer (pH 7.4)
, NAD ⁺ 0.05 ml, 0.1 M ethanol 0.
A reaction solution was prepared by mixing and mixing 1 ml of water and 0.19 ml of water. Next, the reaction solution was cooled to 30 ° C.
, And 0.05 ml of the crude enzyme solution preliminarily heated to 30 ° C. was added and reacted. And 340
The change in absorbance with time at nm was measured, and the rate of increase in absorbance was determined to determine the alcohol dehydrogenase activity (units / mg). 1 μm per minute
The amount of the enzyme that reduces ole NAD ⁺ was defined as 1 unit.

[Presence or absence of β-D-glucan]
To confirm whether D-glucan is present, use high performance liquid chromatography (HPLC) to
D-glucan was detected. Then, the peak of β-D-glucan was recognized, and it was confirmed that the peak was present in the wine. FIG. 3 shows data of wine (Example 7) manufactured using agaricus stake.
It can be confirmed that a peak of -D-glucan exists.
FIG. 4 shows data of commercially available wine (comparative example), and it can be confirmed that there is no peak in the portion indicated by the arrow.

[Fibrinolytic activity] Fibrin plate (5671 m
0.03 ml of wine was sprayed in a circle on the surface of m ² ) (spray area 19.6 mm ² ), and the dissolution area after standing for 1 hour was measured.

[Anti-thrombin activity] The time from the change of fibrinogen to fibrin by the reaction of thrombin to the coagulation (thrombin time) was measured. That is, 0.1 ml of wine was allowed to act on 0.4 ml of a 0.13% by weight fibrinogen solution, and the time required for coagulation was measured.

[0044]

[Table 1]

[0045]

[Table 2]

[0046]

[Table 3]

From the results shown in Tables 1 to 3, the wines produced using basidiomycetes having alcohol dehydrogenase activity (Examples 1 to 12) contain β-D-glucan as a physiologically active substance. You can see that it is included. In contrast, it can be seen that commercially available wine (comparative product) does not contain β-D-glucan. Further, it can be confirmed that the wines of Examples 1 to 12 show fibrinolytic activity. Furthermore, the wines manufactured using oyster mushroom, enokitake mushroom, matsutake mushroom, mustard mushroom, and mannentake mushroom as the basidiomycetes (Examples 1 to 4, 5, 5, and 9 to 12) are compared with commercially available wines (comparative examples) Can also be confirmed to exhibit antithrombin activity.

Next, the antibacterial activity test was performed on the wines of Examples 7 and 8 and Comparative Example as described below. The results are shown in Table 4 below.

[Antibacterial activity test] Black mold (Aspergillus niger, Aspergillus niger) which is a fungal fungus, fungal yeast (Saccharomyces cerevisiae, Saccharomyces cerevisiae), Escherichia coli which is a gram negative bacterium, The antibacterial activity against Bacillus natto, a gram-positive bacterium, was evaluated. Specifically, first, a standard agar medium capable of growing each of the above-mentioned bacteria was kept at 55 ° C., and then each of the bacteria was transplanted, poured into a petri dish with a penicillin cup, and solidified. After solidification, the penicillin cup was removed, wine was added into the hole, and each bacterium was cultured under standard conditions. As a result, those in which an inhibition circle was formed (showing antibacterial activity) were marked with "O", and those in which no inhibition circle was formed (thing showing no antibacterial activity) were marked with "x".

[0050]

[Table 4]

From the results shown in Table 4, the wines manufactured using Agaricus staphy as basidiomycetes (Examples 7 and 8) were as follows:
It can be seen that it has antibacterial activity against black mold, yeast and Escherichia coli. On the other hand, the comparative example product has no antibacterial activity against black mold, yeast, Escherichia coli, and Bacillus natto.

[0052]

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

The use of the basidiomycete has the advantage that wine can be produced not only under anaerobic conditions but also under aerobic conditions. Therefore, the conditions for producing wine can be eased. In addition, when alcohol fermentation is performed using specific basidiomycetes under anaerobic conditions, the conventional manufacturing equipment can be used as it is, and there is an advantage that wine can be manufactured without adding new equipment.

[Brief description of the drawings]

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

FIG. 2 is a flow sheet showing the procedure of the method for producing wine of the present invention.

FIG. 3 is a chart diagram of wine of an example product.

FIG. 4 is a chart of wine of a comparative example product.

FIG. 5 is a flow sheet showing the procedure of a conventional wine production method.

Claims (4)

[Claims] 1. A method for producing wine, wherein in a fermentation step in the production of wine, alcohol fermentation is carried out by basidiomycetes having alcohol dehydrogenase activity. 2. The method according to claim 1, wherein the alcohol fermentation is performed under anaerobic conditions. 3. The method according to claim 1, wherein the alcohol fermentation is performed under aerobic conditions. 4. A wine obtained by the method for producing wine according to claim 1.