JP2003116524A - Useful metabolite obtained by culturing yeast using saccharide culture solution - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a useful metabolite (fermented liquid) by making a cocultivation of a plurality kind of yeasts in a culture solution. SOLUTION: This metabolite(fermented liquid) is obtained by the following process: the plurality kind of yeasts C1 to Cn precultured respectively in tanks 3a to 3n are added to an initial culture tank 1 poured with a culture solution 2 to make an initial cocultivation in the culture solution 2a; after a lapse of a specified time, the cocultivation product is put to aging culture in a relevant tank 9; after completing the aging culture, the product is freed from the yeasts. The metabolite thus obtained is to be used after dilution according to its applications including antioxidation, antimicrobial use and moisturizing use; besides, it is also possible to further raise the efficacy of the metabolite by adding naturally derived components D1, D2 and D3 such as vegetables, fruits and crude drugs to the culture solution.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a useful metabolite using yeast, and particularly, except in the case of yeast of a specific species, a plurality of yeasts having different sugar assimilation properties are co-cultivated in a sugar culture medium. The present invention relates to useful metabolites thus obtained.

[0002]

2. Description of the Related Art Various substances such as preservatives, preservatives, and antioxidants are added to cosmetics and foods. For example, parabens (methylparaben, ethylparaben, butylparaben) are added as preservatives to most cosmetic products. These are effective against a very wide range of microorganisms, but on the other hand, they are so powerful that they are a strong irritant for people with sensitive skin, and in some cases they may cause contact dermatitis, allergic eczema, etc. Become.

Next, various substances are added as preservatives to many foods. For example, various substances such as isobutyl paraoxybenzoate, isopropyl paraoxybenzoate, benzoic acid, sorbic acid and propionic acid are added as preservatives.

Among the above preservatives, there are many substances such as paraoxybenzoic acid which do not exist in nature, and most of the preservatives are synthesized from petroleum as a raw material.

[0005]

Each of the above substances has been nationally approved for use as a preservative and a preservative, and is not highly toxic when used alone or for a short period of time. It is said that there is no. However,
It has been pointed out that if these substances are ingested for a long period of time or if a plurality of substances are combined, they have various adverse effects on the human body such as allergic factors, carcinogenic factors and mutagenicity.

At present, most of the foods, especially processed foods, use the above substances in large amounts as preservatives, and our eating habits focus on such processed foods. As a result, we ingest large amounts of these preservatives on a daily basis, and the possibility of long-term ingestion is increasing.

Further, there is a great possibility that the accumulation of such additives will be inherited by the offspring, resulting in mutation of genes and allergic constitution, and adversely affecting humanity in the future.

Further, foods and cosmetics are deteriorated by oxidation. In particular, lipids are oxidized by oxygen to become lipid peroxides, which deteriorates the quality and deteriorates the flavor. When a person eats the lipids, heartburn, indigestion and the like are adversely affected to the human body.

For these reasons, antioxidants are added to many foods and cosmetics. Conventionally, it has been believed that antioxidants are safe, but the risk of using artificial substances as antioxidants is pointed out, as is pointed out by the carcinogenicity of phenolic antioxidants such as BHA. It is like this.

[0010]

Means for Solving the Problems The present invention is constituted in view of the above-mentioned problems, and by co-culturing a plurality of yeasts (yeast group) having different sugar assimilation properties in a sugar culture solution, Alternatively, by culturing a specific yeast in a single culture or at a ratio close to that of a single culture, a natural and safe metabolite (fermentation liquid) having antioxidative properties, antibacterial properties, moisturizing properties (binding property, viscosity), etc. can be obtained. It is characterized by obtaining.

[0011]

BEST MODE FOR CARRYING OUT THE INVENTION In a sugar culture medium, a plurality of species including a fungus centered on yeast, preferably yeast, more preferably yeast Saccharomyces family, and still more preferably yeast Digosaccharomyces is co-cultivated. Alternatively, one kind of the yeast genus Digosaccharomyces is singly cultivated, or co-cultivation of a yeast strain mainly culturing the yeast Digosaccharomyces is performed.

Components of plants, such as vegetables, fruits, seaweeds, herbs, etc., which can be used as Chinese herbs, animals, plants, minerals, etc., depending on the intended use of the fermented liquor containing useful metabolites finally obtained. It is also possible to add active ingredients such as vitamins, minerals and enzymes into the fermentation broth to increase the added value of the fermentation broth, which is a metabolite.

The co-cultivation is divided into, for example, an initial culture and an aging culture. The initial culture is, for example, about 1 to 7 days, and the aging culture is, for example, about 3 days to 1 year.

The metabolites that have been matured are filtered to obtain a stock solution used for a predetermined purpose. For example, when it is used as a preservative, the effect is sufficiently exhibited by diluting it to 1 to 10% (weight ratio / the concentration is shown below by weight ratio unless otherwise specified) before use.

[0015]

EXAMPLES Examples of the present invention will be described below. Reference numeral 1 in the figure
Is an initial culture tank for performing initial co-cultivation (hereinafter simply referred to as “initial culture”), and a sugar solution containing a saccharide is injected as a culture solution 2 into the initial culture tank 1. As the raw material sugar of the sugar solution, various kinds such as polysaccharides, monosaccharides, oligosaccharides and honey can be used, but it is preferable to use mainly oligosaccharides.

The sugar concentration of the culture solution 2 is about 2% to about 6%.
It can be up to 0%, but it is preferable to use a solution having a high osmotic pressure of 40% to 50%.

As the fungi to be added to the culture solution 2a in the initial culture tank 1, a plurality of fungi having different sugar assimilation properties are selected. Fungi centered on yeast that ferment sugar solution, preferably yeast, more preferably yeast Saccharomyces family, more preferably using a plurality of species including yeast Digosaccharomyces bacterium, this culture Liquid 2a is added (inoculated) and co-cultivated.

The fungus to be co-cultivated is preferably pre-cultured in a sugar solution having a sugar concentration of about 2% to about 40%. In the figure, symbols C1, C2, and Cn represent the types of bacteria to be added, and 3a, 3b, and 3n represent preculture tanks for preculturing these bacteria. The pre-culture liquid containing the pre-cultured fungus is injected into the initial culture tank 1 filled with the culture liquid 2 to add a predetermined bacterium to the culture liquid 2a, and the initial culture is performed.

During this initial culture, temperature adjustment 4, humidity adjustment 5, aeration adjustment 6, sugar concentration adjustment 7, and PH measurement 8 are performed to set the environment according to the conditions of the initial culture.

The conditions of the initial culture are as follows. (1) The sugar concentration of the culture solution 2 is from 2% to 55%. (2) Humidity adjustment 5 is from 30% to 80%. (3) The pH of the culture solution is 3 to 7. (4) Temperature adjustment 4 is from 10 ° C to 35 ° C. (5) The culture time is from 1 to 7 days.

Next, when the initial culture is completed, aging culture is started. In this case, when the plant extract or the like described later is not added, the aging culture may be performed using the initial culture tank 1. In the configuration shown, the case where no additive material such as plants is used will be described first, and then the case where the additive material is used will be described.

After the initial culture, the culture solution 2a is poured into the aging tank 9 as the culture solution 2b, and the aging culture is carried out. In the aging culture, the culturing is carried out under the following conditions so that the co-cultivation of a plurality of types of bacteria is surely advanced, and as a result, useful components are efficiently and stably produced by fermentation. Particularly important points in co-cultivation are the shape of bubbles formed on the upper surface of the culture solution 2b, the pH of the culture solution 2b, the sugar content, and the like.
The temperature adjustment 4, the humidity adjustment 5, the ventilation adjustment 6, and the sugar concentration adjustment 7 are performed while observing these numerical values and conditions.

The conditions of aging culture are as follows. (1) The sugar concentration adjustment 7 of the culture solution 2b is from 20% to 60%. Highly osmolar sugar concentrations of 50% to 65% are preferred. (2) Humidity adjustment 5 is from 30% to 80%. (3) The pH of the culture solution 2b is 4 to 7. (4) Temperature adjustment 4 is from 20 ° C to 35 ° C. It should be noted that it is desirable to adjust the temperature within plus / minus 2 ° C. depending on the bacterial species and raw material. (5) Culture time is from 3 days to 1 year. The number of aging days varies depending on the combination of symbiotic bacteria and the blending of raw materials.

When the aging culture is completed under the above conditions, the removal 11 of the bacteria is carried out. Methods for removing the bacteria include filtration with a filter and sterilization by heating. This culture solution (fermentation solution) is used as the metabolite A after diluting it according to the purpose corresponding to its intended use such as a preservative and a moisturizer. Unlike the case of the metabolite B described below, the metabolite A is a plant extract D1, a drug D2 that can be used as a herbal medicine,
Animal, plant and mineral component D3 is not added.

Next, the case where additives are used in the aging culture process will be described. Depending on the intended use of the metabolites (food, cosmetics, health foods, pharmaceuticals, etc.), vegetables,
Extract D1 of plants such as fruits, seaweeds, and herbs, or D2 that can be used as a herbal medicine, component D of plants and animals, minerals
It is also possible to add 3 or the like and extract active ingredients such as vitamins, minerals, and enzymes into the fermentation broth to make the fermentation broth, which is a metabolite, more effective depending on the above-mentioned use.

In the initial culture tank 1, instead of the sugar solution, an extract D1 of the above-mentioned plants such as vegetables, fruits, seaweeds, herbs, etc., which can be used as a herbal medicine D2, plants and animals, and mineral components D3 Is added to carry out initial culture and aging culture. In this case, when injecting the initial culture solution 2a into the aging culture tank 9, the residue of plants, animal minerals, etc. is roughly filtered by the filter 10 and then transferred to the aging culture tank 9 for aging culture. To do.

When the aging culture is completed by satisfying the conditions of the aging culture described above, the culture solution is extracted from the aging culture tank 9, and after removing the bacteria 11, the metabolite B is used as the final product. obtain. It should be noted that, as described above, the method 11 for removing the bacteria includes filtration with a filter and sterilization by heating.

[0028]

[Test 1] An antiseptic test was conducted using the metabolite (fermentation solution) of the present invention. The test method is as follows.

(Used Metabolite) A fermentation solution obtained by co-cultivation with a plurality of yeasts, that is, a metabolite in FIG. Metabolites classified into A were used. The reason why plants were not added was to eliminate the effect of naturally occurring antibacterial components possessed by plants.

[0030] [Test target fungi] (Target 1)   Mixing of the following 3 kinds of bacteria     E. coli IFO 3972 (Escherichia coli)     Pseudomonas aeruginosa IFO 13275 (Pseudomonas aeruginosa)     Staphylococcus aureus IFO 13726 (Staphylococcus aureus) (Target 2)     yeast     Candida IFO 1594 (Candida albicans) (Target 3)     Mold     Aspergillus niger IFO 4407

[Test Method] The metabolite A of the present invention was inoculated with each of the above three kinds of subjects at a predetermined inoculation concentration, and the temperature was set to 25 ° C.
After culturing for 4 weeks, changes in the number of bacteria were measured. The inoculation concentration was about 10 ⁶ / g only for the above target 1 and 10 for the above targets 2 and 3.
⁵ / g, and the medium for measuring the number of bacteria is SCD only for the above target 1.
LP agar medium, subjects 2 and 3 were GPLP agar medium. Under this culture condition, 10% solution of the metabolite of the present invention, 5%
The tests were carried out using solutions, 1% solutions.

[Test Results] In all cases, the number of bacteria was remarkably reduced, and it was confirmed that the fermentation broth, which is the metabolite of the present invention, has a high antiseptic effect. FIG. 2 shows the test results using a 5% solution, and it can be seen that it is particularly effective for molds.

[0033]

[Test 2] [Water-retaining property (binding property or viscosity), antibacterial property, and antioxidant effect test on livestock products] [Livestock products to be tested] Livestock products that do not contain the metabolite of the present invention are not added Then, a test was carried out by adding the metabolite of the present invention to this non-addition group as a fermentation solution addition group. (Mixing ratio of additive-free section) Pork thigh meat (g): 3000 Pork fat (g): 750 Salt (g): 72 Sugar (g): 18.8 Sodium nitrite (g): 0.563 Ice: 938

(Ratio of Fermented Liquid Addition) The above materials were all in the same proportions as those in the non-addition group.
And 75 ml of the metabolite used in Test 2 was added to the fermented liquid addition section.

(Production of Meat Emulsion) A meat emulsion was produced according to the following procedure using the above-mentioned mixing ratio materials. The cutter used was R-301 manufactured by Stefan.
It is a type. (1) The metabolite of the present invention is added to pig thigh meat (meat temperature of 15 ° C.) (by the way, the above metabolite is not added in the non-addition group). (2) After 15 seconds have passed, the emulsion is covered with a non-ventilated vinyl chloride film to block air (meat temperature 18 ° C.). (3) After 20 minutes, add half amount of water. (4) After 30 seconds, add salt, sugar and half amount of water. (5) Add fat after 30 seconds. (6) After 45 seconds, the meat emulsion was completed (meat temperature 21
C).

(Production of sausage) The above meat emulsion is heated to complete a processed animal product (sausage). (1) About 200 g of the above-mentioned meat emulsion was filled in a fine plus casing (folding diameter: 66 mm) and then steamed at 70 ° C. (2) After confirming that the center temperature of the sausage reached 63 ° C, the sausage was further cooked for 30 minutes. (3) A sample was prepared by boiling, cooling with cold water for 30 minutes, and cooling with a refrigerator (3 ° C.) overnight. The sample was air-wrapped with a non-breathable laminated film and stored at 10 ° C.

[Analysis Items] (1) Water Retention (Binding or Viscosity) 10 g of the meat emulsion was weighed on four wire meshes (15 mesh, 25 cm ² ), inserted into a binding bottle, and a rubber stopper was attached. After that, it was heated in a hot water bath at 75 ° C. for 20 minutes.
After heating, the sample was centrifuged. Centrifuge 2 minutes 30
From 3 seconds to 3 minutes, the rotation speed was increased to 1000 plus or minus 20 rpm, and thereafter, centrifugation was performed at the same rotation speed for 3 minutes. After centrifugation, the amount of the separated liquid was measured and the water retention (binding property) was calculated by the following formula. Binding property (%) = (A−B / A) × 100 Here, A and B are as follows. A: Water content (%) of meat emulsion B: Amount of separated liquid (%)

As a result of measuring the binding property by the above method, the non-addition group was 79.6% and the fermentation solution addition group was 82.7%, and it was confirmed that the binding property was increased by the addition of the metabolite of the present invention. did it.

(2) Peroxide value As shown in FIG. 3, when added, the fermented liquid-added group showed a peroxide value of about 1/2 as compared with the non-added group, and then both of them on the 5th day. After that, the non-addition group rose sharply. On the other hand, the fermentation liquor addition group did not increase thereafter, and the antioxidant effect of the present invention was confirmed.

(3) Antibacterial activity As shown in FIG. 4, almost no increase in bacteria was observed in the non-addition group and the fermentation solution addition group for about one week. However, after 1 week, the number of general viable bacteria increased rapidly in the non-addition group. On the other hand, in the fermented liquid addition group, the number of general viable bacteria did not increase, but rather gradually decreased, and the antibacterial activity against the food of the present invention was confirmed.

Although the efficacy of metabolites to which vegetables, fruits, seaweeds, herbal medicines, etc. have been added has not been tested, the results of the above test show that the addition and addition of these additives will result in higher and higher levels. It is presumed that it will exert the desired effects.

According to a test separately conducted by the inventors,
When the yeast Digosaccharomyces is used for fermentation, even if it is a co-cultivation with only one kind of this fungus or a ratio close to a single culture of this fungus, it is close to the case of the co-cultivation of the fungus shown in Examples. It was confirmed that it is possible to obtain a metabolite having an effect.

[0043]

EFFECTS OF THE INVENTION As specifically shown above, the present invention is basically a metabolite of co-cultivation of a eubacterium with a focus on saccharides and yeast, which is different from conventional naturally derived products. It is a substance that can be obtained inexpensively and stably without having to go through an extraction and purification step that is inefficient and lacks in stability, and has high antibacterial properties, antioxidant properties, moisturizing properties, and the like.

Since the above metabolites do not contain artificially synthesized substances such as drugs, they are very safe. By diluting and using the metabolites of the present invention, pharmaceuticals, cosmetics, foods, dietary supplements can be prepared. It can be widely used as a safe and safe natural antibacterial agent, antioxidant, moisturizer, etc. for feed, fertilizer, other water and environmental improvement according to its purpose.

In addition, plants, seaweed extracts, those that can be used as herbal medicines, animals and plants, mineral components, etc. are appropriately selected and added, and these active ingredients are extracted into metabolites without any special steps. The effect of these metabolites can be further enhanced by these additives.

[Brief description of drawings]

FIG. 1 is a process chart showing an example of a process for obtaining a useful metabolite (fermentation solution) according to the present invention.

FIG. 2 is a graph showing the results of testing the antiseptic activity of the metabolite of the present invention (fermentation solution) for mixed bacteria, yeast, and mold.

FIG. 3 is a graph showing the results of testing the changes in the peroxide value of the processed livestock products by dividing them into a non-addition group and a fermentation liquid addition group to which the metabolite of the present invention (fermentation liquid) was added.

FIG. 4 shows processed animal products divided into an additive-free group and a fermentation solution-added group to which the metabolite of the present invention (fermentation solution) has been added, and the results of testing the antibacterial properties of both by the change in the number of general viable bacteria It is a graph shown.

[Explanation of symbols]

1 Initial culture tank 2, 2a, 2b culture solution 3a, 3b, 3n Preculture tank 4 Temperature adjustment 5 Humidity adjustment 6 Ventilation adjustment 7 sugar content adjustment 8 PH adjustment 9 Aging culture tank 10 filters Removal of 11 bacteria C1, C2, Cn co-cultivated bacteria D1 plant extract What can be used as D2 herbal medicine D3 Animal and plant components

   ─────────────────────────────────────────────────── ─── Continued front page    (72) Inventor Yuji Tojima             Stocks 1-5 Azabu 10-4 Minato-ku, Tokyo             Company Plans Board Souei F term (reference) 4B042 AC06 AD03 AK16                 4B065 AA80X BB22 BC50 CA41

Claims (6)

[Claims] 1. A useful metabolite obtained by culturing a bacterium using a sugar culture solution, which is obtained by co-culturing a plurality of eubacteria having different sugar assimilation in a culture solution containing a saccharide. 2. The useful metabolite obtained by culturing a bacterium using the sugar culture solution according to claim 1, wherein the eubacterium is yeast. 3. The useful metabolite obtained by culturing a bacterium using the sugar culture solution according to claim 2, wherein the yeast is a yeast belonging to the family Saccharomyces setae. 4. One of the yeasts belonging to the family Saccharomyces setae belongs to the genus Digosaccharomyces, and is obtained by culturing a bacterium using a sugar culture solution according to claim 3. Useful metabolites. 5. A useful metabolite obtained by culturing a bacterium using a sugar culture medium, which is obtained by culturing a yeast digosaccharomyces alone in a culture medium containing a saccharide. . 6. A naturally occurring component such as vegetables, fruits, seaweeds, animals and plants used as a herbal medicine, or a mineral or the like or an extract containing the component is added to a culture solution containing a saccharide, and these naturally derived components are added. 6. A useful metabolite obtained by culturing a bacterium using the sugar culture solution according to claim 1 or 5, which is obtained by extraction into the culture solution.