JP5876214B2 - Method for producing lactic acid bacteria symbiotic culture extract - Google Patents

Description

The present invention relates to a method for producing a lactic acid bacteria symbiotic culture extract obtained by culturing and extracting a group of lactic acid bacteria.

  For example, health food processed from sweet potato stems and leaves (Japanese Patent Laid-Open No. 2006-306851), health food processed from kuzuhana (Japanese Patent Laid-Open No. 2006-262888), and enteric bacteria improving agent extracted from bark (Japanese Patent Laid-Open No. 2007-308373) No.), beverages containing sphingomyelin (Japanese Unexamined Patent Publication No. 2007-112793), fermented acerola fermented acerola fruit (Japanese Unexamined Patent Publication No. 2005-253463), or an interface containing sphingomyelin as an active ingredient A drug for enhancing Lon production (Japanese Patent No. 3519749) has been developed.

JP 2006-308551 A JP 2006-262888 A JP 2007-308373 A JP 2007-112793 A JP 2005-253463 A Japanese Patent No. 3519749

  According to the “New Influenza Action Plan”, which was decided by the Ministry of Education, Culture, Sports, Science and Technology in September 2006, “To close universities and refrain from going out as much as possible, and to communicate with universities and students during the closure period. It was stipulated that the method should be clarified and sufficient guidance should be given for student life during the closing period.

  Recently, influenza has been transformed into a new type of influenza instead of avian flu in Mexico, and has been raging throughout the world, including Japan, and WHO has called for caution by accrediting it as Phase 6 of its heyday.

  It is also true that stress is likely to accumulate due to excessive information, and gastrointestinal diseases such as irritable colitis and traveler's diarrhea, or diseases such as allergic dermatitis occur frequently. .

  In view of the above, even if it is a dietary supplement, I think that it will contribute to the suppression of influenza, the suppression of the said disease, etc., and it will help health promotion.

  An object of the present invention is to provide a lactic acid bacteria symbiotic culture extract obtained by culturing and extracting a group of lactic acid bacteria, which can contribute to the suppression of influenza and the above-mentioned diseases.

In order to achieve the above object, the method for producing a lactic acid bacterium symbiotic culture extract according to the present invention contains vitamin H (biotin), sphingomyelin, vitamin B12 as main components, and folic acid, Na (sodium) as other components. ), Cl (chloride) , K (potassium), IP (inorganic phosphorus), Ca (calcium), Mg (magnesium), Fe (iron), Cu (copper), Zn (zinc), ethanol, and lactic acid. The contained medium is selected , lactic acid bacteria are cultured in the selected medium, vitamin H (biotin) 3.7 ng / ml, sphingomyelin 0.2 mg / ml, vitamin B12 156 pg / ml, folic acid 0.3 ng / ml, Na (sodium) 5 mEq / l, Cl (chloride) 21 mEq / l, K (potassium) 1.0 mEq / l, IP (inorganic phosphorus) 0 .3 mEq / l, Ca (calcium) 0.3 mEq / l, Mg (magnesium) 0.5 μg / g, Fe (iron) 5 μg / g, Cu (copper) less than 0.1 μg / l, Zn (zinc) It is characterized by extracting an extract solution containing less than 1 μg / g, ethanol 0.2% v / v, and lactic acid 2.58% as components.

  According to the present invention, as a result of clinical trials, the effect of suppressing allergic reaction, activation of NK cells, immune function, irritable colitis, traveler's diarrhea, traveler's diarrhea, ulcerative colitis, food poisoning, etc. Effects such as improvement of organ system, allergic dermatitis, immune function, anti-aging effect (anti-aging), food texture improvement, shelf life (preservation effect), diabetes prevention, etc. can be obtained.

It is a figure which shows the result of the trial 1 using the lactic-acid-bacteria symbiotic culture extract which concerns on embodiment of this invention. It is a figure which shows the result of the trial 2 using the lactic-acid-bacteria symbiotic culture extract which concerns on embodiment of this invention. It is a figure which shows the result of the trial 3 using the lactic-acid-bacteria symbiotic culture extract which concerns on embodiment of this invention. It is a figure which shows the result of the trial 4 using the lactic-acid-bacteria symbiotic culture extract which concerns on embodiment of this invention. It is a figure which shows the result of the trial 6 using the lactic-acid-bacteria symbiotic culture extract which concerns on embodiment of this invention. It is a figure which shows the result of the trial 7 using the lactic-acid-bacteria symbiotic culture extract which concerns on embodiment of this invention. It is a figure which shows the result of the trial 8 using the lactic-acid-bacteria symbiotic culture extract which concerns on embodiment of this invention. It is a figure which shows the result of the trial 8 using the lactic-acid-bacteria symbiotic culture extract which concerns on embodiment of this invention. It is a figure which shows the result of the trial 8 using the lactic-acid-bacteria symbiotic culture extract which concerns on embodiment of this invention.

  Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings.

The method for producing a lactic acid bacteria symbiotic culture extract according to an embodiment of the present invention contains vitamin H (biotin), sphingomyelin, and vitamin B12 as main components, and folic acid, Na (sodium), Cl ( as other components ). Chloride) , K (potassium), IP (inorganic phosphorus), Ca (calcium), Mg (magnesium), Fe (iron), Cu (copper), Zn (zinc), ethanol, and a medium containing lactic acid as components. Lactic acid bacteria are cultured in the selected medium, vitamin H (biotin) 3.7 ng / ml, sphingomyelin 0.2 mg / ml, vitamin B12 156 pg / ml, folic acid 0.3 ng / ml, Na (sodium) 5 mEq / l, Cl (chloride) 21 mEq / l, K (potassium) 1.0 mEq / l, IP (inorganic phosphorus) 0.3 mEq / l , Ca (calcium) 0.3 mEq / l, Mg (magnesium) 0.5 μg / g, Fe (iron) 5 μg / g, Cu (copper) less than 0.1 μg / l, Zn (zinc) less than 1 μg / g It is characterized by extracting an extract containing g, ethanol 0.2% v / v, and lactic acid 2.58% as components.

In the embodiment of the present invention, the main components contained in the extract obtained by culturing and extracting the selected lactic acid bacteria group were examined. In the measurement, a plurality of the extract solutions were prepared as samples. As a result of arbitrarily selecting one sample extract and examining its components, the following results were obtained.
Vitamin H (biotin) 3.7 ng / ml
Sphingomyelin 0.2 mg / ml
Vitamin B12 156 pg / ml
Could be detected as the main component.
Besides this ingredient,
Folic acid 0.3 ng / ml
Was included. In addition, Na (sodium), Cl (salt), K (potassium), IP (inorganic phosphorus), Ca (calcium), Mg (magnesium), Fe (iron), Cu (copper), Zn (zinc), although in trace amounts. Ethanol was also included. An example of the content of these contained components contained in the extract was as follows.
Na (sodium) 5 mEq / l
Cl (chloride) 21 mEq / l
K (potassium) 1.0 mEq / l
IP (inorganic phosphorus) 0.3 mEq / l
Ca (calcium) 0.3 mEq / l
Mg (magnesium) 0.5 μg / g
Fe (iron) 5 μg / g
Cu (copper) less than 0.1 μg / l
Zn (zinc) less than 1 μg / g
Ethanol 0.2% v / v
Lactic acid 2.58%

Next, the contents of the main components contained in the remaining plurality of sample extracts were similarly examined. Based on the results, the contents of vitamin H (biotin), sphingomyelin, and vitamin B12, which are the main components contained in the lactic acid bacteria symbiotic culture extract according to this embodiment, are included in the following ranges. Met. The phenomenon in which the content of the main component is in a range is considered to be caused by subtle changes in conditions in the environment for culturing lactic acid bacteria and the process of extracting the extract.
Vitamin H (biotin) Minimum value 3.3 ng / ml to maximum value 4.1 ng / ml
Sphingomyelin minimum value 0.18mg / ml to maximum value 0.22mg / ml
Vitamin B12 minimum 140.0 pg / ml to maximum 171.6 pg / ml
For folic acid, the maximum value is 0.3 ng / ml. Depending on the extract of the sample, there may be a trace amount, and the content of folic acid is in the range of 0.3 ng / ml to 0.03 ng / ml. Is desirable.

The inventor extracts an extract containing the following main components, conducts various clinical trials using these lactic acid bacteria symbiotic culture extracts, and the effectiveness of the lactic acid bacteria symbiotic culture extract according to the embodiment of the present invention. , Examined the efficacy.
(Sample 1)
Vitamin H (biotin) 3.3 ng / ml
Sphingomyelin 0.18mg / ml
Vitamin B12 140.0pg / ml
(Sample 2)
Vitamin H (biotin) 3.7 ng / ml
Sphingomyelin 0.2mg / ml
Vitamin B12 156pg / ml
(Sample 3)
Vitamin H (biotin) 4.1 ng / ml
Sphingomyelin 0.22mg / ml
Vitamin B12 171.6pg / ml

  As for folic acid, the Ministry of Health, Labor and Welfare recommends that pregnant women should take 400 μg of folic acid daily. Since the lactic acid bacteria symbiotic culture extract according to the embodiment of the present invention contains vitamin H (biotin) and vitamin B12 as main components and folic acid is taken together with these, the effectiveness and efficacy of folic acid is sufficient. I think it can be predicted.

  Hereinafter, the results of the following clinical trials 1 to 8 using the lactic acid bacteria symbiotic culture extract of samples 1 to 3 described above will be described.

  Lactobacillus symbiotic culture extract according to an embodiment of the present invention is an extract obtained by culturing and extracting a group of lactic acid bacteria, improving digestive system such as hypersensitivity colitis, traveler's diarrhea, ulcerative colitis, food poisoning, Not only is it expected to have various effects such as allergic dermatitis, immune function, anti-aging effect (anti-aging), food texture improvement, shelf life (antiseptic effect), diabetes prevention, etc. Even when applied to animals, it is expected to have an effect of intestinal regulation, appetite enhancement, infectious disease improvement and anti-inflammatory by immune enhancement.

(Clinical trial 1)
Therefore, it was examined by measuring IFN-γ and IL-4 in blood how drinking of lactic acid bacteria symbiotic culture extract has an effect on immune balance.

  Moreover, the placebo effect was examined by classifying into a group for drinking lactic acid bacteria symbiotic culture extract and a group for drinking a pseudo drug.

(Clinical trial method)
The subjects and methods of the experiment were explained in advance to the subjects (a total of 6 people, one 22-year-old man and five women), and the study was conducted with the consent of the subjects.
Of the 6 subjects, 3 were drunk with lactic acid bacteria symbiotic culture extract, the other 3 were imitation drugs, 5 ml each day for 1 week each morning. Then, blood was collected every day at 10 am to obtain serum. Subjects were not informed in advance of which one was consumed to observe the placebo effect.

  In order to examine the immune balance, the cytokines IFN-γ and IL-4 in the blood were measured, and a clinical trial was conducted to determine whether there was a placebo effect. In the measurement, blood is collected, serum is then separated, the separated serum is stored at −20 ° C., and the cytokine IFN-γ is analyzed by a flow cytometry method using a cytokine measurement kit Flow Cytomix (BECKMAN COULTER). And IL-4 were measured.

(effect)
FIG. 1 shows the relationship between the number of days taken and the amount of change (%) in Th1 / Th2 as a result of the clinical trial. The horizontal axis of FIG. 1 represents the number of days of drinking, and the vertical axis represents the amount of change (%) in Th1 / Th2. Moreover, the solid line of FIG. 1 has shown the case where the said lactic acid bacteria symbiotic culture extract is drunk, and the dotted line has shown the case where a pseudo drug is drunk, respectively.

  As is clear from the clinical trial results shown in FIG. 1, it can be seen that when the lactic acid bacteria symbiotic culture extract is drunk, the rate of change of IFN-γ / IL-4 (Th1 / Th2) increases day by day. On the other hand, when the simulated drug was drunk, no significant change was observed in the amount of change in IFN-γ / IL-4 (Th1 / Th2). From the clinical trial results in FIG. 1, it can be seen that drinking the lactic acid bacteria symbiotic culture extract has the effect of increasing Th1 / Th2 and that there was no placebo effect.

(Discussion)
Helper T cells (Th cells) are divided into Th1 cells and Th2 cells based on the pattern of cytokine secretion. IFN-γ measured this time is a main cytokine produced by Th1 cells, has a strong macrophage activation action, and is effective in eliminating pathogens.
In contrast, IL-4 is a cytokine that produces Th2 cells and has the effect of promoting the production of IgE antibodies. IL-4 suppresses the activation of macrophages and suppresses the function of Th1 cells.
Thus, the immune function is adjusted by the balance of action between Th1 and Th2.

As shown in FIG. 1, as a result of the trial of drinking the lactic acid bacteria symbiotic culture extract, the increase in IFN-γ / IL-4 every day indicates activation of IFN-γ (Th1 cells), or It is thought that suppression of IL-4 (Th2 cell) is performed.
From the above clinical trial results, it was confirmed that drinking the lactic acid bacteria symbiotic culture extract is effective in suppressing allergic reactions with the lactic acid bacteria symbiotic culture extract.

(Clinical trial 2)
Next, infection and defense (defense) experiments of pathogenic Escherichia coli and Salmonella by oral administration of the lactic acid bacteria symbiotic culture extract to mice were conducted. In this experiment, a lactic acid bacteria symbiotic culture extract solution having a concentration of 5% was orally administered to mice, and the efficacy of the lactic acid bacteria symbiotic culture extract solution against infection with pathogenic Escherichia coli (O-18) and Salmonella (S. typhimurium ATCC14028). Observations were made using mice.

(experimental method)
Six mice were grouped into four groups, and each group was allowed to freely absorb a 5% concentration lactic acid bacteria symbiotic culture extract according to the following schedule, and then the mice were administered Salmonella. However, Salmonella was administered on the same day.
(1) Group 1 From 5 days before administration of Salmonella, 5% lactic acid bacteria co-culture extract was administered.
(2) Group 2 Salmonella and 5% lactic acid bacteria symbiotic culture extract were administered.
(3) Group 3 From 5 days after administration of Salmonella, 5% lactic acid bacteria co-culture extract was administered.
(4) Group 4 Only Salmonella was administered.

The above experimental results are shown in FIG.
In group 1, 6 mice did not die from Salmonella by administration of 5% lactic acid bacteria symbiotic culture extract despite administration of Salmonella.
In group 2, one out of 6 mice died from Salmonella infection.
In group 3, of 6 mice, 2 mice died from infection with Salmonella.
In group 4, 3 out of 6 mice died from infection with Salmonella.
All the mice that died were mice whose BW (body weight) fell below 20%. In addition, one of the mice in group 1 also had a BW (weight) of 20% or less, but survived from there.

(Discussion)
Based on the above results, the mortality rate of mice was 50% when only Group 4 Salmonella was administered. However, in the group administered with 5% lactic acid bacteria symbiotic culture extract, the mortality rate of mice by administration of Salmonella is 0% in group 1, 16.7% in group 2, 33.3% in group 3. The earlier the time when the 5% concentration lactic acid bacteria symbiotic culture extract was administered, the lower the mortality rate.
From this result, it is considered that the administration of the lactic acid bacterium symbiotic culture extract has preventive and therapeutic effects against the action in the digestive tract of pathogenic Escherichia coli (O-18) and Salmonella causing food poisoning. It is done.
As a result of examining the lactic acid bacteria symbiotic culture extract, it is considered that the lactic acid bacteria symbiotic culture extract is effective in preventing and improving various symptoms including the digestive system, and one of its mechanisms. It is thought that this is due to the increase of good bacteria in the intestine.

(Clinical trial 3)
Next, the above-mentioned lactic acid bacteria symbiotic culture extract was administered to a subject with a tumor, and the result of image diagnosis was obtained.

(Method)
March 20, 2008 Administration of 20 ml / d of the lactic acid bacteria symbiotic culture extract started March 24, 2008 Administration of 50 ml / d of the lactic acid bacteria symbiotic culture extract April 20, 2008 50 ml / d of the lactic acid bacteria symbiotic culture extract d administration May 3, 2008 75 ml / d administration of the lactic acid bacteria symbiotic culture extract

(result)
In addition, pain score shall be shown as score 0-10 before the start of administration (March 20, 2008), 0 when healthy and intact, and 10 when most painful. , Described by patient subjectivity.
March 6, 2008 Left iliac pain score 8
March 20, 2008 Left iliac pain score 10
March 24, 2008 Left iliac pain score 8
April 20, 2008 Left iliac pain score 6
May 3, 2008 Left iliac pain score 3
May 14, 2008 Left iliac pain score 1-2

  As described above, by administering the lactic acid bacteria symbiotic culture extract, left iliac pain was suppressed every day, and finally left iliac pain was reduced from “8” to “1-2”. As a result of the image diagnosis of bone scintigraphy shown in FIG. 3, a shrinking tendency was seen in the tumor nest of the left iliac part as indicated by an arrow.

(Clinical trial 4)
Next, a clinical trial was conducted on the temporal change of NK cell activity due to administration of the lactic acid bacteria symbiotic culture extract. In this clinical trial, subjects who complained of body temperature 39 °, headache, sore throat, joint pain, and muscle pain were allowed to drink 10 ml of the lactic acid bacteria symbiotic culture extract, and the activity of NK cells over time was measured. The result is shown in FIG.

  In FIG. 4, the horizontal axis represents time, and the vertical axis represents NK cell activity (%). For comparison, the activity of healthy NK cells is indicated by a dotted line.

  When the subject's body temperature reached 39 ° C and the subject complained of headache, sore throat, joint pain, and muscle pain, 10 ml of the lactic acid bacterium symbiotic culture extract was administered to the subject, and NK cells with time passed. The degree of activity was measured.

As is clear from FIG. 4, the NK cell activity increased to 60% when 2 hours passed after the subject was infected with influenza and ingested the lactic acid bacteria symbiotic culture extract. The activity level of NK cells showed a tendency to decrease, and after 6 hours, the activity level of NK cells of the subject became the same value as that of healthy subjects.
According to the results of this clinical trial, when the lactic acid bacteria symbiotic culture extract is administered immediately upon infection with influenza, the degree of NK cell activity is increased within a short time (after 2 hours of drinking), and then (for 6 hours). After), it turns out that it returns to a healthy state. Therefore, the lactic acid bacteria symbiotic culture extract is considered to be effective in preventing influenza.

(Clinical trial 5)
5 ml of the lactic acid bacteria symbiotic culture extract was administered to a 22-year-old female medical student, who was the subject, to examine changes in constipation and intestinal microflora before and after administration. The subject had constipation and defecation once a week.
The results are shown below.
Before administration After administration (2 months later)
Status of constipation Once / week → Once / day [Intestinal flora]
Anaerobic bacteria 71.4% → 46.5%
Lactic acid bacteria 0.0% → 0.6%
Bifigus no detection → 2.0%

As is clear from the above results, the anaerobic bacteria in the intestine decreased from 71.4% to 46.5%, the lactic acid bacteria increased from 0.0% to 0.6%, and the bifigus bacteria was 2.0%. Rose to%. As a result, the stool was in a state of constipation once a week before administration, but the stool was improved once a day after administration.
This also shows that constipation is improved by administering the lactic acid bacteria symbiotic culture extract.

(Clinical trial 6)
Next, 5 mice of the same day and 1 year of age are divided into 2 groups, and a lactic acid bacteria symbiotic culture extract of 5% concentration is administered to the mice of group 1, and the lactic acid symbiotic culture extract is administered to the mice of group 2. The course was observed without administration.

  FIG. 5 shows the state of mice in Group 2 bred without administration of the lactic acid bacteria symbiotic culture extract and the state of mice administered with the lactic acid symbiotic culture extract.

When the skin of group 2 mice shown in the upper left of FIG. 5 is observed, a bald spot is seen in a part thereof. However, when the skin of the group 1 mouse shown in the lower right of FIG. 5 is observed, the mice observed with the administration of the lactic acid bacterium symbiotic culture extract having a concentration of 5% do not have a wrinkle portion, and the skin does not appear. Stay healthy.
Although this phenomenon is unknown medically (biologically), the life span of the mouse is about 24 months, and in particular, the mouse used this time has been recognized to be generalized as the death phase approaches ( Aging phenomenon).
As a result of administering a 5% concentration lactic acid bacteria symbiotic culture extract to group 1 mice, the group 1 mice seem to have an anti-aging effect due to the synergistic effect of folic acid contained in the lactic acid symbiotic culture extract. Can be considered.

  In addition, among the mice in the group administered with the 5% concentration lactic acid bacteria symbiotic culture extract, the last surviving mouse was 3 months after the death of the mice that survived without administering the 5% concentration lactic acid bacteria symbiotic culture extract. Survived in good health. Three months for a mouse corresponds to about 10 years in terms of human age. Also from this fact, it was found that administration of the lactic acid bacteria symbiotic culture extract has the effect of anti-aging effect (anti-aging).

  Moreover, from the anti-aging effect confirmed in the clinical trial 6, it is considered that there is an effect and efficacy even when folic acid is contained. In that case, the content of the folic acid is desirably 0.3 ng or less (ng / ml) in 1 ml of the extract.

(Clinical trial 7)
Next, a clinical trial was conducted to determine whether or not diabetes prevention is exhibited by using the lactic acid bacteria symbiotic culture extract according to this embodiment as a blood glucose level adjusting substance. In the clinical trial method, 5 healthy subjects aged 21 to 22 years were selected as subjects, and a 75 g oral glucose tolerance test (75 gOGTT) was performed (Trailan G75 g tolerance test). The results are shown in FIGS. 6 (a), (b) and (c), respectively.

  In the clinical trial method, 5 healthy subjects aged 21 to 22 years old were selected as subjects, and a 75 g oral glucose tolerance test was simultaneously ingested with a lactic acid bacterium symbiotic culture extract as a blood glucose regulator, and blood glucose levels were measured over time. At that time, a 75 g oral glucose tolerance test was performed by changing the intake of the lactic acid bacteria symbiotic culture extract. In FIG. 6, the horizontal axis indicates time, and the vertical axis indicates blood glucose level (mg / dl). Furthermore, in FIG. 6, the bar line connected with ◆ shows the change in blood glucose level when only Tralan G75g was ingested, and the bar line connected with ■ indicates that Tralan G75g and lactic acid bacteria co-culture extract were ingested simultaneously. This shows the change in blood glucose level.

FIG. 6 (a) shows the results of measuring changes in blood glucose level when 2 ml of lactic acid bacteria symbiotic culture extract was simultaneously ingested as a blood glucose regulating substance in the 75 g oral glucose tolerance test.
As is clear from FIG. 6 (a), when 30 to 60 minutes have elapsed, the blood glucose level when ingesting Tralan G75g and the lactic acid bacteria symbiotic culture extract at the same time is the blood glucose level when Toran G75g alone is ingested. It is lower than the value. When 60 to 80 minutes have passed, there is no change in the blood glucose level when Toleran G75g and lactic acid bacteria symbiotic culture extract are ingested at the same time, and the blood sugar level tends to be higher than that with Toleran G75g alone. there were. When 80 minutes passed and 150 minutes passed, the blood glucose level when taking Tralan G75g and the lactic acid bacteria symbiotic culture extract simultaneously was lower than the blood sugar level when taking Tralan G75g alone. . And when 180 minutes passed, the blood glucose level returned to the value before ingestion.

FIG. 6B shows the results of measuring changes in blood glucose level when 5 ml of lactic acid bacteria symbiotic culture extract was simultaneously ingested as a blood glucose regulating substance in the 75 g oral glucose tolerance test.
As is clear from FIG. 6 (b), when 30 to 60 minutes have elapsed, the blood glucose level when ingesting Toraylan G75g and the lactic acid bacteria symbiotic culture extract at the same time is the blood sugar level in ingesting Toraylan G75g alone. It is lower than the value. When 60 to 80 minutes have passed, there is no change in the blood glucose level when Toleran G75g and Lactobacillus symbiotic culture extract are ingested at the same time, but the blood sugar level is lower than that with Toleran G75g alone. Show. When 80 minutes passed and 150 minutes passed, the blood glucose level when taking Tralan G75g and the lactic acid bacteria symbiotic culture extract simultaneously was lower than the blood sugar level when taking Tralan G75g alone. . And when 180 minutes passed, the blood glucose level returned to the value before ingestion.
It is clear from FIG. 6 (b) that the blood glucose level when taking up Tralan G75g and 5 ml of lactic acid bacteria symbiotic culture extract simultaneously is lower than the blood sugar level when taking Tralan G75g alone, regardless of the passage of time. You can see that

FIG. 6 (c) shows the results of measuring the change in blood glucose level when 10 ml of lactic acid bacteria symbiotic culture extract was simultaneously ingested as a blood glucose regulating substance in the 75 g oral glucose tolerance test.
As is clear from FIG. 6 (c), when 30 to 60 minutes have elapsed, the blood glucose level when ingesting Toraylan G75g and the lactic acid bacteria symbiotic culture extract at the same time is the blood sugar level in ingesting Toraylan G75g alone. It is lower than the value. When 60 to 80 minutes have passed, there is no change in the blood glucose level when Toleran G75g and Lactobacillus symbiotic culture extract are ingested at the same time, but the blood sugar level is lower than that with Toleran G75g alone. Show. When 80 minutes passed and 150 minutes passed, the blood glucose level when taking Tralan G75g and the lactic acid bacteria symbiotic culture extract simultaneously was lower than the blood sugar level when taking Tralan G75g alone. . And when 180 minutes passed, the blood glucose level returned to the value before ingestion.
It is clear from FIG. 6 (c) that the blood glucose level when taking up Tralan G75g and 10 ml of lactic acid bacteria symbiotic culture extract at the same time is lower than the blood sugar level when taking Tralan G75g alone, regardless of the passage of time. You can see that The tendency of this change in blood glucose level was the same as in FIG.

(Discussion)
From the results shown in FIG. 6, it was shown that the lactic acid bacteria symbiotic culture extract has a function of lowering the blood glucose level. Furthermore, the blood glucose level inhibitory action was strongly recognized by administering 5 ml or more of lactic acid bacteria symbiotic culture extract. From the results of the above clinical trials, it was found that the lactic acid bacteria symbiotic culture extract according to the present embodiment exhibits the effect of preventing diabetes.

(Clinical trial 8)
Next, a clinical trial was conducted on the texture and shelf life (generation of mold; antiseptic effect) when the lactic acid bacteria symbiotic culture extract according to the present embodiment was used as an additive for bread. The results are shown in FIGS.

As a clinical trial method, a bread to which the lactic acid bacteria symbiotic culture extract according to the present embodiment was added and an additive-free bread were produced using a household bread maker. Each bread was cut in half, tasted half, and the other half was put in a plastic bag for comparison.
The photographs on the right side of FIGS. 7 to 9 show the bread produced according to the instructions attached to the household bread maker. The photographs on the left side of FIGS. 7 to 9 show the bread produced based on the above description using 10% water of the lactic acid bacteria symbiotic culture extract according to this embodiment.
Moreover, the numbers appended to the photographs in FIGS. 7 to 9 indicate the elapsed days from the date of manufacture.

For the clinical trial, breads of January 19, 2010 were produced, and immediately after production, they were aseptically placed in plastic bags and stored at 20 ° C., and the progress was observed.
At the stage of January 25, 2010, two molds were found at the position indicated by a circle in the additive-free bread, as is apparent from the photograph on the right side of the top row in FIG.
On the other hand, in the case of the bread to which the lactic acid bacterium symbiotic culture extract shown in the left photograph of FIG. 6 is added, two molds are present at the position indicated by ○ at the stage of January 28, 2010 as shown in FIG. It's been found.
As shown in FIG. 9, at the stage of February 1, 2010, in the additive-free bread shown in the photograph on the right side of FIG. 9, the mold had expanded to an area of about 1/3. On the other hand, in the bread to which the lactic acid bacteria symbiotic culture extract shown in the left photograph of FIG. 9 was added, mold was hardly expanded.
As shown in FIG. 9, at the stage of February 2, 2010, in the additive-free bread shown in the photograph on the right side of FIG. 9, mold was clearly enlarged almost entirely. On the other hand, in the bread to which the lactic acid bacteria symbiotic culture extract shown in the left photograph of FIG. 9 was added, the mold was thinly expanded to about ½.

(Discussion)
The textures of the additive-free bread produced on January 19, 2010 and the bread added with the lactic acid bacterium symbiotic culture extract of this embodiment were compared. In the case of the bread to which the lactic acid bacterium symbiotic culture extract of this embodiment was added, there was a mochi and moist feeling that was not found in the additive-free bread, and there was a flavor.
Furthermore, as is apparent from the results shown in FIGS. 7 to 9, mold was generated after production in the case of bread to which the lactic acid bacteria symbiotic culture extract of this embodiment was added, although it was 9 days after production. On the other hand, in the case of additive-free bread, it was the sixth day after production, and the bread of the bread added with the lactic acid bacteria symbiotic culture extract of this embodiment was prolonged by about 50% compared to the additive-free bread.

(Conclusion)
By adding the lactic acid bacteria symbiotic culture extract of this embodiment to bread, the texture and shelf life (preservative effect) were clearly improved. This suggests that the lactic acid bacterium symbiotic culture extract of the present embodiment can be expected to have the same effect as an additive for other foods as a natural food additive.

  Based on the results of the above clinical trials, the lactic acid bacteria symbiotic culture extract according to the embodiment of the present invention contains biotin (vitamin H), sphingomyelin, and vitamin B12 as main components, and biotin (vitamin H) in 1 ml of the extract. 3.3 to 4.1 ng (ng / ml), sphingomyelin in the extract, 0.18 to 0.22 mg (mg / ml) in 1 ml of the extract, vitamin B12 in 140 ml in the extract. Including 0 to 171.6 pg (pg / ml), the results of the trial indicate that it is a dietary supplement, but is associated with irritable colitis, traveler's diarrhea, traveler's diarrhea, ulcerative colitis , Improvement of digestive system such as food poisoning, allergic dermatitis, immune function, various effects such as anti-aging effect (anti-aging), food texture improvement, shelf life (antiseptic effect), diabetes One in which it is possible to demonstrate the efficacy of such proof.

Next, examples of using the lactic acid bacteria symbiotic culture extract according to the present embodiment as an animal supplement and contributing to the improvement of symptoms will be listed.
(1) An example of cat's mouth sores that caused a stomatitis and appetite was abolished. Appetite was abolished and I went to a veterinary clinic. .
When a 6-year-old Japanese cat was orally administered 2 ml of the lactic acid bacteria symbiotic culture extract according to this embodiment, appetite began after about 1 hour, all the findings were the same as normal, and the behavior was 3, I became 4 years old.
Since then, 1 ml daily has been administered orally and maintained in good condition for 2 years.
(2) Use of intractable chronic diarrhea for Akita dogs
In Akita prefecture with intractable chronic diarrhea that lasted for more than 1 year, nothing was detected by microbiological examinations (bacteria, fungi, parasites), and symptomatic treatment was performed but no improvement was observed.
When the lactic acid bacteria symbiotic culture extract according to this embodiment was given orally at 2 ml per day, diarrhea stopped within a few days.
(3) Use for cats with intractable external auditoryitis (topical)
A 5-year-old Japanese cat who went to a veterinary clinic for many years, had a bad odor from his ears, had pain, and hardly improved. When the lactic acid bacteria symbiotic culture extract according to this embodiment was diluted about 50 times and the local area was wiped with a cotton swab or the like, it improved in several days and the symptoms disappeared.
(4) Use for cats with intractable external auditoryitis (systemic)
A 6-year-old Japanese cat had the same situation as Case 3. When 1 ml of the lactic acid bacteria symbiotic culture extract according to this embodiment was orally administered, it improved in several days and the symptoms disappeared.
(5) Use of colds for cats A 5-year-old Japanese cat with a body temperature of about 39 ° C, sneezed, thought to have a cold without appetite.
When 2 ml of the lactic acid bacteria symbiotic culture extract according to this embodiment was orally administered, appetite appeared after about 30 minutes, and it was eaten as usual, and then recovered smoothly.

  The lactic acid bacteria symbiotic culture extract according to this embodiment is considered to have an intestinal regulating action, an appetite enhancing action, an infectious disease ameliorating action and an anti-inflammatory action by enhancing immunity even when applied to animals as in humans. Expected to help with the disease.

  Although the present invention is a dietary supplement, it not only contributes to prevention and improvement of pathogenic diseases such as new influenza that are prevalent in this term, but also irritable colitis, traveler diarrhea, traveler diarrhea, Improvement of digestive system such as ulcerative colitis, food poisoning, allergic dermatitis, immune function, efficacy such as anti-aging effect (anti-aging), food texture improvement, shelf life (antiseptic effect), prevention of diabetes In addition to exerting such effects, it is considered that even when applied to animals, it has an intestinal regulating action, an appetite enhancing action, an infectious disease improving action and an anti-inflammatory action by enhancing immunity.

Claims (1)

Vitamin H (biotin), sphingomyelin, and vitamin B12 are included as the main components, and folic acid, Na (sodium), Cl (chloride) , K (potassium), IP (inorganic phosphorus), Ca (other components) A medium containing calcium, Mg (magnesium), Fe (iron), Cu (copper), Zn (zinc), ethanol, and lactic acid as components is selected , lactic acid bacteria are cultured in the selected medium, and vitamin H ( (Biotin) 3.7 ng / ml, sphingomyelin 0.2 mg / ml, vitamin B12 156 pg / ml, folic acid 0.3 ng / ml, Na (sodium) 5 mEq / l, Cl (chloride) 21 mEq / l, K (potassium) 1.0 mEq / l, IP (inorganic phosphorus) 0.3 mEq / l, Ca (calcium) 0.3 mEq / l, Mg (magnesium 0.5 μg / g, Fe (iron) 5 μg / g, Cu (copper) less than 0.1 μg / l, Zn (zinc) less than 1 μg / g, ethanol 0.2% v / v, lactic acid A method for producing a lactic acid bacteria symbiotic culture extract comprising extracting an extract containing 58% as a component.

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