JP3276929B2 - Concentrated beverage with anti-inflammatory and anti-infective effects - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to the beverage concentrate with anti-inflammatory action and infection protective action, more particularly, Bull
Gall bacilli, granulobacilli, acidophiles, lactococci, yeast
Five beneficial bacteria Liquid concentrate filtrate co raw culture-obtained culture broth, to a beverage concentrate with anti-inflammatory action and infection protective action.

[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, Bulgarian rods
Bacteria, balance granules bacilli, from 嗜酸bacilli, Lactococcus, filtrate five beneficial bacteria obtained by co-live culture broth of yeast, amino acid by concentration method, the effective ingredients such as vitamins and minerals It has been found that a concentrated solution containing well and having a high nutritional value can be obtained, and that this concentrated solution is suitable as a health drink. And, through further studies, they found that this concentrated beverage had an inflammation inhibitory effect and an infectious disease protective effect. The present invention is based on such findings. That is, the concentrated beverage having an inflammation-suppressing action and an infectious disease-protecting action according to the present invention is a Bulgarian rod.
Bacteria, a granule bacillus,嗜酸bacilli, Lactococcus, concentrate itself filtrate five beneficial bacteria were obtained by co-viable culture broth of yeast, additives Artificial been used at all Absent.
And <br/> beneficial bacteria used in the production of beverage concentrate according to the present invention, may be mentioned also that shown in Table 1.

[0005]

[Table 1]

[0006] The concentrated beverage according to the present invention is a Bulgarian rod.
Bacteria, granules bacilli,嗜酸bacilli, Lactococcus, is obtained by concentrating the filtrate five beneficial bacteria were obtained by co-viable culture broth of yeast, as groupings of these beneficial bacteria Can be exemplified, for example, as shown in Table 2.

[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]

In the concentrated beverage according to the present invention, as amino acids, in addition to essential amino acids such as threonine, valine, lysine and leucine, almost 20 kinds of amino acids such as glutamic acid, aspartic acid and arginine could be confirmed. .

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 caused swelling of the paw of the rats for 5 to 18 days after inoculation, and the rats were inoculated with the concentrated beverage of the present invention for 1 hour to 2 days a day for 5 consecutive days.
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
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, metastasis of inflammation to the forelimbs was suppressed (4/5 in the case where 0.5% CMC was orally administered, as compared to the case where 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 (4/5 the same as when 0.5% CMC was orally administered, when the concentrated beverage according to the present invention was orally administered) 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 includes Bulgarian bacilli, granular bacilli, and acidophila.
Bacteria, Lactococcus, be one obtained by concentrating the filtrate of a microorganism fermentation product is a culture broth obtained by co-live cultured five beneficial bacteria yeast, artificial additives It is not used at all and is suitable as a health drink.

[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 a Bulgarian bacillus, a granulobacillus, a peracid bacillus, a lactic acid sphere.
Bacteria, produced by concentrating the filtrate five beneficial bacteria were obtained by co-viable culture broth yeast. Optimally, soymilk from soybeans is used as the culture medium. Milk is commonly used for cultivation of lactic acid bacteria.Natural drinks that do not use any artificial additives in consideration of the non-uniformity of milk, its aging, and the use of chemicals in its production process. In view of this, it is most preferable to use soymilk, which is a natural food and completely pesticide-free soybean (from Iowa), as a culture medium. 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) 3rd step (manufacture of special soymilk culture medium) After removing fat from soybeans, adding distilled water and boiling for 70-110 minutes, add salt, trisaccharide and beneficial bacteria to the filtered soymilk filtrate. 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 placed in a thermostat. And culture at 39 ° C for 48-50 hours. (5) Fifth step (medium volume adaptation 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) Candida albicans (ATCC10231) was aerated at 37 ° C for 16 hours using a Sabouraud liquid medium (Difco, Detroit, Michigan, USA) (hereinafter abbreviated as SB medium) as a culture medium. Phase-growing bacteria were obtained. 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). Viable mold count of inoculum is 550nm
Was measured based on the turbidity measured by the above method, and was accurately measured by inoculating a serial dilution on an SB agar plate. Male ICR
Mice (5 weeks old, about 24 g) were 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) The Sabouraud liquid medium was used as a culture medium to prepare Candida albi.
Cans (ATCC10231) was subjected to aeration culture at 37 ° C. for 16 hours to obtain logarithmically growing bacteria. 0-10% mortality in normal mice,
An inoculum containing the amount of mold expected to produce 70% mortality in immunosuppressed mice 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. Concentrated beverage according to the present invention and azimexone (Azimexone) is PBS (pH 7.4)
And diluted. Male ICR mice (4 weeks old, about 20
g), 0.8 ml of test compound or PBS was administered by intraperitoneal administration (ip) on days 1, 3 and 5, and the immunosuppressant cyclophosphamide (30 mg / Kg) was administered orally (po).
They were given on days 4 and 6. On day 7, the mouse was 0.2 m
l of Candida albicans diluted inoculum was inoculated and then surviving animals were scored for 10 days after inoculation.

(In vivo effect) In the immunomodulation test, 10
Using an ICR mouse, 1% and 0.1% of the concentrated beverage according to the present invention or 100% by intraperitoneal administration (ip) was given to each mouse.
mg / Kg of 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 (1% dose was administered) In this case, 70 out of 100 animals died, and when administered in the 0.1% dose, 72 out of 100 animals died). The mortality rate with azimexone was 82% (41/50 deaths).

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.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ identification code FI A61K 35/74 A23L 2/00 F (58) Field surveyed (Int.Cl. ⁷ , DB name) A23L 1/28-1 / 30 A61K 35/72-35/74

Claims (1)

(57) [Claims] (1) Bulgarian bacilli, granular bacilli, acid bacilli, milk
Acid cocci, five beneficial bacteria co raw culture-obtained culture solution of the filtrate concentrated beverage with anti-inflammatory action and infection protective action, which is a concentrate of yeast.