JP2000125810A - Concentrated beverage having suppressing action on inflammation and protecting action on infectious disease - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a concentrated beverage having suppressing actions on inflammations, protecting actions on infectious diseases, immunomodulating actions and recovering actions and suitable for a healthy beverage, etc., by including a yeast fungus and a lactic acid bacterium. SOLUTION: This concentrated beverage comprises (A) a yeast fungus such as a yeast and (B) a lactic acid bacterium such as Bacillus bulgaricus. The concentrated beverage is preferably a concentrate of a filtrate of a culture solution prepared by symbiotic culture of plural species of useful microorganisms selected from the yeast fungi and lactic acid bacteria. A soybean milk from completely organic guaranteed soybeans is preferably used as a raw material.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a concentrated beverage having an inflammation-inhibiting action and an infectious disease-protecting action, and more particularly to a concentrated beverage comprising yeast and lactic acid bacteria having an inflammation-inhibiting action and an infectious disease-protecting action. is there.

[0002]

2. Description of the Related Art Various drink products such as pharmaceutical drinks, sports drinks and soft drinks are commercially available. In many of these beverage products, artificial food additives are added in order to enhance nutritional value, palatability or preservability.

[0003]

However, these artificial food additives are not necessarily said to have no adverse effects on health, and in some cases, harm to the human body becomes a social problem. The present inventor has intensively studied to provide a beverage produced by using a microorganism fermentation product that does not use artificial additives.

[0004]

As a result, amino acids and vitamins are concentrated by a concentration method from the filtrate of a culture solution obtained by co-cultivating a plurality of beneficial bacteria selected from yeast and lactic acid bacteria in a specific combination. It has been found that a concentrated liquid having a high nutritional value containing active ingredients such as minerals and minerals in a well-balanced manner can be obtained, and that this concentrated liquid is suitable as a health drink. And as we continue our research,
It has been found that this concentrated beverage has an inflammation inhibitory effect and an infectious disease protective effect. The present invention is based on such findings. That is, the concentrated beverage according to the present invention having an inflammation-suppressing effect and an infectious disease-protecting effect comprises yeast and lactic acid bacteria. More specifically, it comprises a concentrated filtrate of a culture solution obtained by co-culturing a plurality of beneficial bacteria selected from yeast and lactic acid bacteria in a specific combination. The beneficial bacteria (yeast and lactic acid bacteria) used in the production of the concentrated beverage according to the present invention include those shown in Table 1.

[0005]

[Table 1]

[0006] The concentrated beverage according to the present invention is obtained by concentrating a filtrate of a culture solution obtained by co-culturing a plurality of beneficial bacteria selected from yeast and lactic acid bacteria in a specific combination. Examples of the grouping of a plurality of types of beneficial bacteria include the cases shown in Table 2, for example.

[0007]

[Table 2]

Analysis of the concentrated beverage according to the present invention reveals that protein, phosphorus, iron, calcium, thiamine (vitamin
B ₁ ), riboflavin (vitamin B ₂ ) and so on. Table 3 shows an example of the analysis result.

[0009]

[Table 3]

[0010] In the concentrated beverage according to the present invention, nearly 20 amino acids such as glutamic acid, aspartic acid and arginine could be confirmed as essential amino acids such as threonine, valine, lysine and leucine. .

Incidentally, it has been found that the concentrated beverage according to the present invention has an inflammation-suppressing effect and an infectious disease-protecting effect. The operation will be described in detail with data. The inflammation referred to in the present invention is a concept including various acute and chronic inflammations such as arthritis and allergic reactions. Complete Freund's adjuvant (0.3mg sterile M.tubercul in 0.1ml light mineral oil)
adjuvant arthritis test was performed using rats inoculated in the hind paws with a suspension of osteoarthritis. Inoculation with complete Freund's adjuvant resulted in swelling of the paw of the rats for 5 to 18 days after inoculation.
Oral administration was carried out at a concentration of 100%. In addition, 0.5% CMC (carboxyl methyl cellulose), hydrocortisone
(30 mg / Kg) was orally administered to rats in the same manner. Table 4 shows the results.

[0012]

[Table 4]

As is evident from Table 4, swelling of the rat's foot can be effectively reduced by orally administering the concentrated beverage according to the present invention. Inhibition of paw swelling was observed both in the acute phase (Days 0-5, p <0.05, t-test) and in the late phase (Days 14-18, p <0.05).
In contrast, hydrocortisone was effective in the acute phase (p <0.01, t-test), but had no effect in the late phase. On the other hand, inflammation metastasis from hindpaw to forefoot is 0.5% CM
When C. was orally administered, it was 4/5, whereas when the concentrated beverage according to the present invention was orally administered, it was 2/5 (see P in Table 4). Inflammation metastasis of 0.5
Oral administration of% CMC is 5/5,
When the concentrated beverage according to the present invention was orally administered, the ratio was 1/5 (see T in Table 4), indicating that the concentrated beverage according to the present invention suppressed the transfer of inflammation to both parts. I understand. When hydrocortisone was orally administered, the metastasis of inflammation to the forelimbs was suppressed (in contrast to 4/5 when 0.5% CMC was orally administered, the same as when the concentrated beverage according to the present invention was orally administered 2/5). 5, see P in Table 4), but could not be suppressed by metastasis of inflammation to the tail. 1/5, see T in Table 4).

[0014] Carrageenan-induced footpad edema test was performed on rats to which 0.1 ml of carrageenan (1% suspension) was administered into the hind toes. The administration of carrageenan caused swelling of the hind paws of the rats 3 hours later, and the rats were given a single oral administration of the concentrated beverage of the present invention at a concentration of 100% one hour before the administration of carrageenan. Similarly, distilled water (20 ml / Kg) and aspirin (150 mg / Kg) were orally administered to rats.
Table 5 shows the results.

[0015]

[Table 5]

As is apparent from Table 5, when the concentrated beverage according to the present invention is orally administered, an anti-inflammatory effect is observed.

Next, a passive dermal Analylaxis test was performed using rats sensitized 24 hours before with an intra-skin reactant (IgE) anti-ovalbumin serum (0.5 ml). The skin of this rat caused an allergic reaction after 30 minutes with ovalbumin, but the concentrated beverage according to the present invention was given to this rat.
Ovalbumin administration at 100% and 50% respectively 1
A single oral dose was given hours before. Also, distilled water (20ml / Kg),
Aspirin (3 mg / Kg) was similarly administered orally to rats. Table 6 shows the results.

[0018]

[Table 6]

As is clear from Table 6, when the concentrated beverage according to the present invention is orally administered at a concentration of 100%, it is recognized that the allergic reaction caused by ovalbumin is effectively inhibited. In addition, rats were treated with a skin reactant (Ig
E) Sensitized 24 h before with anti-ovalbumin serum (0.5 ml). The inhibition rate (%) was calculated by intravenous administration of ovalbumin (1 mg) plus Evans blue (5 mg), and recording the skin wheal reaction 30 minutes later.

Histamine diphosphate (30 μl in 0.05 ml)
g) Rats inoculated with g) were subjected to passive skin Analylaxis test. 10 minutes after inoculation, wheal was observed on the skin of the rat, and the concentrated beverage according to the present invention was applied to this rat.
Oral administration at a 100% concentration. Also, distilled water (20ml / Kg),
Cyproheptadine (3 mg / Kg) was similarly orally administered to rats. Table 7 shows the results.

[0021]

[Table 7]

As is clear from Table 7, the concentrated beverage according to the present invention does not act on histamine. The inhibition rate (%) was calculated by recording the decrease in skin wheals 10 minutes after inoculation of histamine diphosphate (30 μg in 0.05 ml) into the skin.

As is clear from the above, it can be seen that the concentrated beverage according to the present invention has various acute and chronic inflammation suppressing effects such as arthritis and allergic reactions. The concentrated beverage according to the present invention is obtained by concentrating a filtrate of a microbial fermentation product, which is a culture solution obtained by co-cultivating a plurality of beneficial bacteria selected from yeast and lactic acid bacteria in a specific combination. It is suitable for health drinks.

[0024]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the concentrated beverage according to the present invention will be described in more detail with reference to specific examples. The concentrated beverage according to the present invention is composed of yeast and lactic acid bacteria, and more specifically, a culture solution obtained by co-culturing a plurality of beneficial bacteria selected from yeast and lactic acid bacteria in a specific combination. Produced by concentrating the filtrate. Optimally, soymilk from soybeans is used as the culture medium. Milk is generally used for culturing lactic acid bacteria.
Considering time-dependent variability, use and mixing of drugs and the like in the production process, and further, from the viewpoint of a natural drink that does not use any artificial additives, it is a natural food as a culture medium,
It is best to use soy milk from 100% organically guaranteed soybeans (Iowa). Further, the concentrated beverage according to the present invention is produced by heating and sterilizing live bacteria after culture is completed to remove the cells, and concentrating the filtrate of the culture solution from which the cells were separated to about 1/15. Is preferred.

Next, an example of a method for producing a concentrated beverage according to the present invention will be described. As one example, Japanese Patent No. 1962512
No. (Japanese Patent Publication No. 6-95914). (1) First step (manufacture of special agar culture medium) To a purified natural agar, add soup, salt, and beneficial bacteria culture solution obtained from beef and kelp that have been roasted with distilled water to obtain a special agar culture medium To manufacture. (2) Second step (pure cultivation of beneficial bacteria) Beneficial bacteria of each kind are transplanted to the special agar culture medium produced in the first step, which is placed in a thermostat and cultured at 39 ° C for 48 to 50 hours. (3) Third step (manufacture of special soymilk culture medium) After removing fat from soybeans, adding distilled water and boiling for 70 to 110 minutes, culture the filtered soymilk filtrate with salt, trisaccharide and beneficial bacteria The liquid is added to produce a special soymilk culture medium. (4) Fourth step (pure cultivation of beneficial bacteria in special soymilk culture medium) The beneficial bacteria purely cultured in the second step are transplanted for each type into the special soymilk culture medium produced in the third step and put in a thermostat. And culture at 39 ° C for 48-50 hours. (5) Fifth step (medium volume conditioned culture of beneficial bacteria) Purely cultured beneficial bacteria produced in the fourth step are divided into groups on the special soymilk culture medium obtained in the third step, and transplanted for each group. For 48-50 hours at 39 ° C. (6) Sixth step (industrial symbiotic culture of beneficial bacteria) In 13 liters of special soymilk culture medium obtained in the third step,
According to the group produced in the fifth step, live cells conditioned and cultured are collectively transplanted and co-cultivated at 40 ° C. for 100 to 120 hours. (7) Step 7 (Stopping and Concentrating Culture) The probiotic culture produced in Step 6 is heated at 100 ° C. for 30 minutes to stop the growth of viable bacteria, and then the viable bacteria are separated and removed. The filtrate obtained is concentrated to about 1/15 (removing 93-96% of water). (8) Eighth Step (Aging) The concentrated solution obtained in the seventh step is allowed to stand at 17 ° C. for 3 months or more to mature. The product obtained by the above steps is sealed in a container and subjected to legal heating to obtain a concentrated beverage product.

The concentrated beverage according to the present invention can be produced through such a process, and can be summarized as a fermentation product of yeast and lactic acid bacteria. It can also be said that it is a natural health drink without artificial additives. As described above, this concentrated beverage has an inflammation-suppressing action, and drinking it can suppress various acute and chronic inflammations such as arthritis and allergic reactions.

The concentrated beverage according to the present invention has an immunosuppressive effect. In using the mouse model
In vivo (in vivo test), a test on immunosuppression of the concentrated beverage according to the present invention showed a 20% increase in survival rate as compared with the control of infected animals, and an immunosuppression test by cyclophosphamide treatment Showed a 30% increase in survival. From these results, it was found that the concentrated beverage according to the present invention has an immunosuppressive effect. Hereinafter, the immunosuppression test will be briefly described.

(Immune adjustment test) Sabouraud liquid medium (Difc
o, Detroit, Michigan, USA (S
B medium), Candida albican
s (ATCC10231) was subjected to aeration culture at 37 ° C. for 16 hours to obtain log phase growing bacteria. An inoculum containing an amount of mold that could be expected to have a 90-100% mortality was prepared by diluting the culture overnight with a phosphate buffer solution (hereinafter abbreviated as PBS). The number of viable molds of the inoculum was determined based on the turbidity at 550 nm, and was accurately determined by inoculating serial dilutions on SB agar plates. Male ICR mice (5 weeks old, approx.
24 g) was injected intravenously with 0.2 ml of the diluted inoculum. The inoculum ranged from 1.5 × 10 ⁷ to 2.0 × 10 ⁷ CFU per mouse.

(Immune recovery test) Candida albicans (ATCC10231) was used as a culture medium with the above Sabouraud liquid medium.
Aeration culture was performed at 16 ° C. for 16 hours to obtain logarithmic growth bacteria.
0-10% mortality in normal mice, 70 in immunosuppressed mice
An inoculum containing the amount of mold expected to have a% mortality was prepared by diluting the culture overnight with PBS. The number of viable molds of the inoculum was determined based on the turbidity at 550 nm, and was accurately determined by inoculating serial dilutions on SB agar plates. Male ICR mice (4 weeks old, about 20 g) were injected intravenously with 0.2 ml of the diluted inoculum. The inoculation volume was 6 × 10 ⁶ to 8 × 10 ⁶ CFU per mouse. The concentrated beverage according to the present invention and azimexone were solubilized and diluted with PBS (pH 7.4). Intraperitoneal administration to male ICR mice (4 weeks old, about 20g)
0.8 ml of test compound or PBS by (ip)
Administered on days 3 and 5, the immunosuppressant cyclophosphamide (3
0 mg / Kg) was given on days 2, 4, and 6 by oral administration (po). On day 7, the mice received 0.2 ml of Candida albi
Cans diluted inoculum was inoculated, and surviving animals were recorded for 10 days after inoculation.

(In vivo effect) In the immunomodulation test, 10 ICR mice were used for each administration group, and 1% and 0.1% of each mouse were intraperitoneally administered (ip) according to the present invention. Concentrated beverage or 100mg / Kg azimexone
(Azimexone) was administered, and the number of deaths in 10 experiments was accumulated every day and totaled. In the PBS control group,
The mortality rate was 89% (89 out of 100 animals died), but decreased to about 70% when the concentrated beverage according to the present invention was administered (100% when the 1% dose was administered). 70 of the 100 animals died, or 72 of 100 died when given the 0.1% dose). 82% mortality when receiving azimexone
(41 of 50 died).

In the immune recovery test, 10 ICR mice were used for each administration group, and 1%
And 0.1% concentration of the concentrated beverage according to the present invention or 100 mg / Kg of azimexone were individually intraperitoneally administered (ip), and the number of deaths in eight experiments was accumulated every day, and the total was counted. In the control group administered with PBS and cyclophosphamide, the mortality rate of 75% (60 of 80 animals died) was reduced to 35% and 43% when the concentrated beverage according to the present invention was administered (1%). When the concentration was administered, 28 out of 80 animals died, and when the 0.1% concentration was administered, 34 out of 80 animals died). The mortality rate in the azimexone group was 50% (40 of 80 animals died).

As described above, in the immunomodulation test, the concentrated beverage according to the present invention showed a 20% increase in the survival rate as compared with the control of infected animals, and the recovery test against the immunosuppression by cyclophosphamide treatment. Showed a 30% to 40% increase in survival. As is clear from the above, it can be seen that the concentrated beverage according to the present invention has an immunoregulatory and recovery effect. That is, it has an effect of adjusting and restoring immunity against mold infection when the immune function is normal and when it is in a suppressed state, so that it is possible to protect against various infections or opportunistic infections.

[0033]

EFFECT OF THE INVENTION Since the concentrated beverage of the present invention has an inflammation-suppressing action, drinking it can not only suppress various acute and chronic inflammations such as arthritis and allergic reactions, but also immunomodulation. Since it has a recovery effect, it can also protect against various infections or opportunistic infections.

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[Procedure amendment]

[Submission date] November 20, 1998 (1998.11.
20)

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] 0009

[Correction method] Change

[Correction contents]

[0009]

[Table 3]

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61P 31/00 A61K 35/74 G A61K 35/74 35/78 J 35/78 A23L 2/00 F F term (Reference) 4B017 LC03 LG08 LK21 LP01 LP02 LP05 4B018 LB08 LE05 MD81 MD86 ME07 ME14 MF06 MF13 4C087 AA01 AA02 BC11 BC56 BC58 BC61 BC64 BC70 MA02 MA17 NA05 NA14 ZA96 ZB08 ZB11 ZB13 ZB35 ZC21 ZC22 4C014 AD1712 ZA96 ZB08 ZB11 ZB13 ZB35 ZC21 ZC22

Claims (2)

[Claims] 1. A concentrated beverage comprising an yeast and a lactic acid bacterium, which has an inflammation inhibitory action and an infectious disease protective action. 2. The inflammation-suppressing action and infection according to claim 1, which is a concentrate of a filtrate of a culture solution obtained by co-culturing a plurality of beneficial bacteria selected from yeast and lactic acid bacteria in a specific combination. Concentrated beverage with protective effect.