Composition containing lactic acid bacteria production substance, method for producing equol, and method for producing composition containing lactic acid bacteria production substance

Abstract

To provide fermentation extracts of lactic acid bacteria comprising products produced by lactic acid bacteria, which exert in the body bactericidal effect against so-called bad bacteria as well as anti-aging effect without using chemicals (synthetic chemical substances) by adding a mixture to the extract.SOLUTION: The composition of lactic acid bacteria products using a lactic acid bacteria fermentation extract comprising products produced by lactic acid bacteria contains brown sugar, okara (bean curd), or lactobacillus fermentation extract derived from soybeans, contains more than 22% by weight of plum extract, a rosacea plant, contains ionized apatite with at least natural biogenic or mineral-derived hydroxyapatite and citric acid, contains cherry blossom extract, contains at least one of deep sea water minerals, Great Salt Lake minerals or lactic acid bacteria, and reduced fermented fulvic acid containing fulvic acid, and fermented with hydrogen and deep sea water.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a lactic acid bacteria production substance-containing composition using a lactic acid bacteria fermentation extract comprising a lactic acid bacteria production substance, a method for producing equol, and a method for producing a lactic acid bacteria production substance-containing composition. [Background technology]

[0002] The usefulness of lactic acid bacteria has been known for a long time, and they are used, for example, to solve problems in the oral cavity. Patent Document 1 discloses a technique in which lactic acid bacteria of the genus Leuconostoc are contained in an oral composition to induce coaggregation of major oral bacteria that form oral biofilms. This technique aims to prevent and treat oral diseases by allowing lactic acid bacteria to adhere to the oral mucosa and settle in the oral cavity, thereby efficiently inhibiting the growth of pathogenic bacteria present in the oral cavity.

[0003] Patent Document 2 also discloses a technique for a microorganism or a fragment thereof as an oral care agent for preventing tartar. This technique relates to a composition containing a microorganism or a fragment thereof for reducing mutans streptococci. Such a composition can be used, for example, in an oral care composition for preventing dental caries or for preventing tartar or bad breath.

[0004] Patent Document 3 discloses an inducer of antimicrobial peptides that is safe and has no side effects. In this inducer, the lactic acid bacteria may be ingested as live bacteria or in the form of a material containing live bacteria obtained by fermenting the lactic acid bacteria. It is believed that by ingesting this inducer of endocrine secretion of antimicrobial peptides, the antimicrobial peptides are induced in the body, and it is expected that the improvement of immunity due to various infection defenses, anti-aging, and extension of healthy lifespan can be achieved. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] JP 2010-53062 A [Patent Document 2] International Publication No. WO2012 / 100991 [Patent Document 3] International Publication No. WO2015 / 087919 Summary of the Invention [Problem to be solved by the invention]

[0006] As mentioned above, the benefits of lactic acid bacteria are known, but it has not been fully elucidated how lactic acid bacteria fermentation extract, which is made up of substances produced by lactic acid bacteria, should be combined with other substances to exert a bactericidal effect against so-called bad bacteria or to exert the function of breaking down substances that may cause health problems.

[0007] The present invention has been made in view of the above circumstances, and aims to provide a composition containing a lactic acid bacteria production substance that can exert a bactericidal effect against so-called "bad bacteria (such as Streptococcus mutans, Porphyromonas gingivalis, Pasteurella pylori, Staphylococcus aureus, Clostridium perfringens, Candida, etc.; the same applies below) in the body and an anti-aging effect (anti-glycation, anti-oxidation, skin care) without using chemical substances (synthetic chemical substances) by adding a mixture to a lactic acid bacteria fermentation extract. Candida is generally known as an opportunistic bacteria that is neither good nor bad, but it is a type of mold that rapidly increases in the oral cavity and intestines once the body's immune system weakens, causing oral candidiasis and leaky gut syndrome, and it is said that it is not easy to suppress its proliferation. [Means for solving the problem]

[0008] (1) In order to achieve the above object, the present invention takes the following measures: That is, the lactic acid bacteria production substance-containing composition of the present invention is a lactic acid bacteria production substance-containing composition using a lactic acid bacteria fermentation extract consisting of a lactic acid bacteria production substance, and is characterized in that it contains a lactic acid bacteria fermentation extract derived from brown sugar, soybean pulp, or soybean, and also contains 22% or more by weight of plum extract, which is a plant of the Rosaceae family.

[0009] (2) In the composition containing a lactic acid bacteria production substance of the present invention, the lactic acid bacteria fermentation extract contains 377 (mg / L) to 566 (mg / L) of peptides.

[0010] (3) The lactic acid bacteria production substance-containing composition of the present invention further contains at least one of deep sea water, water-soluble silicon, ceramide, hyaluronic acid, eggshell membrane peptide, nacre pearl powder, and soybean isoflavones including equol or daidzein.

[0011] (4) The composition containing a lactic acid bacteria production substance of the present invention is further characterized by containing 0.03 to 5% by weight of at least one or more of the following: extract of a plant of the Rosaceae family, hibiscus extract, cranberry extract, pomegranate extract, olive extract (leaves, fruit, flower), camellia extract (leaves, fruit, flower), perilla fruit extract, tea fruit extract, or soybean extract.

[0012] (5) The composition containing a lactic acid bacteria production substance of the present invention is characterized by further containing natural biologically-derived or mineral-derived hydroxyapatite and ionized apatite containing at least citric acid.

[0013] (6) The composition containing a lactic acid bacteria production substance of the present invention is characterized by further containing silk hydrogen pearl powder, which is a combination of silk powder and hydroxyapatite with hydrogen and deep sea water.

[0014] (7) The composition containing a lactic acid bacteria production substance of the present invention is also characterized by further containing curcumin and pea sprout extract.

[0015] (8) The composition containing a lactic acid bacteria production substance of the present invention is characterized by further containing at least one of rose petal powder, grape seed powder, hibiscus powder, and cranberry extract.

[0016] (9) The composition containing a lactic acid bacteria production substance of the present invention is characterized by further containing a lactic acid fermentation barley extract and a passion flower extract.

[0017] (10) The composition containing a lactic acid bacteria production substance of the present invention is characterized by containing at least fulvic acid out of fulvic acid or humic acid, and at least reduced and fermented fulvic acid out of deep-sea water minerals or Great Salt Lake minerals which have been reduced and processed with hydrogen, which further contains at least reduced-processed deep-sea water minerals or lactic acid bacteria.

[0018] (11) The composition containing a lactic acid bacteria production substance of the present invention is also characterized by further containing coconut shell activated carbon.

[0019] (12) The composition containing a lactic acid bacteria production substance of the present invention is characterized by further containing a combination of a red orange extract, a pineapple enzyme, a papaya enzyme, hydrogen and deep sea water.

[0020] (13) The composition containing a lactic acid bacteria production substance of the present invention is also characterized by further containing hydrocharcoal, which is a combination of at least one of activated charcoal and edible charcoal, inulin, hydrogen and deep sea water.

[0021] (14) In the composition containing a lactic acid bacteria production substance of the present invention, the hydrocharcoal further contains powder of Kagome kelp and citric acid. In addition to Kagome kelp, seaweed-derived materials such as Akamoku, Mekabu, Ganiashi, and seaweed dulse may be used.

[0022] (15) The composition containing a lactic acid bacteria production substance of the present invention is also characterized by further containing chlorogenic acid.

[0023] (16) The composition containing a lactic acid bacteria production substance of the present invention is characterized by further containing multiple types of oligosaccharides, lactic acid bacteria, and green papaya extract.

[0024] (17) The composition containing a lactic acid bacteria production substance of the present invention is characterized in that it further contains at least one of astaxanthin, an extract of Tricholoma matsutake, Coprino, Pleurotus ulmoides, and Antrodia camphorata in addition to the red pine bark extract powder.

[0025] (18) In addition, in the composition containing a lactic acid bacteria production substance of the present invention, the water-soluble silicon is characterized in that it comprises at least one of plant-derived silicon, mineral-derived silicon, and water-soluble silicon (hydrated silica or hydrated silicic acid such as metasilicic acid or orthosilicic acid) derived from hot springs, spring water, lava, or rocks in the Fuji, Hakone, Kirishima, and Sakurajima regions.

[0026] (19) The method for producing equol of the present invention is characterized in that it comprises adding soybean isoflavones including daidzin or daidzein under specified conditions to the composition containing a lactic acid bacteria production substance described in (1) above.

[0027] (20) Furthermore, the method for producing a composition containing a lactic acid bacteria production substance of the present invention is a method for producing a composition containing a lactic acid bacteria production substance using a lactic acid bacteria fermentation extract consisting of a lactic acid bacteria production substance, and is characterized by including at least the steps of: mixing a lactic acid bacteria fermentation extract derived from brown sugar, soybean pulp, or soybean with 22% or more by weight of plum extract, which is a plant of the Rosaceae family, to produce a mixture; and fermenting the mixture using hydrogen and deep sea water. Effect of the Invention

[0028] According to the present invention, it is possible to obtain an effective bactericidal effect against so-called "bad bacteria" in the body and an anti-aging effect without using chemical substances (synthetic chemical substances). [Brief description of the drawings]

[0029] [Figure 1]FIG. 1 is a graph showing the effect of deep sea water powder on the proliferation of bacteria in commercially available yogurt A. [Diagram 2] FIG. 1 shows the effect of deep sea water powder on the proliferation of bacteria in commercially available yogurt B. [Diagram 3] This is a graph showing the effect of adding deep sea water powder on the proliferation of Lactobacillus gasseri JCM1131, known as an intestinal lactic acid bacterium. [Figure 4] FIG. 1 is a diagram showing an overview of the mechanism of reduced fermenting lactic acid bacteria. [Figure 5A] FIG. 1 is a graph showing an increase in good bacteria and a decrease in bad bacteria. [Figure 5B] FIG. 1 is a diagram showing the functions of lactic acid bacteria fermentation extract (Biogenics). [Figure 6] FIG. 1 shows the results of verification of bioapatite (registered trademark: derived from natural living organisms and low crystallinity) and KT-11 lactic acid bacteria using immune cells. [Figure 7] 1 is an electron microscope photograph showing the results of a test on remineralization. [Figure 8] 1 is an electron microscope photograph showing the results of a test on remineralization. [Figure 9] 1 is an electron microscope photograph showing the results of a test on remineralization. [Figure 10] FIG. 1 is a diagram illustrating the mechanism of glycation. [Figure 11] FIG. 1 is a diagram showing a model of a "food chain." [Figure 12] FIG. 1 shows a model of the fate of fulvic acid. [Figure 13] FIG. 1 is a diagram showing the effects of hydrogen water. [Figure 14] FIG. 1 is a diagram showing a model of the fate of water-soluble silicon. [Figure 15] FIG. 1 shows the results of a test on the synergistic effect between "KS melt lactic acid bacteria production substance (raw material that serves as the base for reduced fermentation lactic acid bacteria)" and "KT-11 lactic acid bacteria HP." [Figure 16] FIG. 1 shows how silicon content in human aorta changes with age. [Figure 17]FIG. 1 is a graph showing a comparison of collagen production rates (relative ratios assuming that no additive is 100%) between types 2 and 3. [Figure 18] FIG. 1 is a graph showing a comparison of collagen production rates (relative ratios assuming that no additive is 100%) between types 2 and 3. [Figure 19] FIG. 1 is a graph showing the effect of bathing in hydrogen water on increasing the moisture content of the stratum corneum of the skin. [Figure 20] FIG. 1 is a graph showing the effect of bathing in hydrogen water on increasing the moisture content of the stratum corneum of the skin. [Figure 21] FIG. 1 shows the effect of a hydrogen bath on moisture loss due to dry skin. [Figure 22] FIG. 1 shows the results showing hydroxyl radical scavenging activity. [Diagram 23] FIG. 1 shows the results showing hydroxyl radical scavenging activity. [Figure 24] FIG. 1 shows the results showing hydroxyl radical scavenging activity. [Figure 25A] FIG. 13 is a graph showing a comparison between product 2 and product 3 in terms of hyaluronic acid production rate (relative ratio assuming no addition as 100%) in a test group containing 31.25 μg / mL collagen. [Figure 25B] FIG. 13 is a graph showing a comparison between product 2 and product 3 in terms of hyaluronic acid production rate (relative ratio assuming no addition as 100%) in a test group containing 125 μg / mL collagen. [Figure 25C] FIG. 13 is a graph showing a comparison between product 2 and product 3 in terms of hyaluronic acid production rate (relative ratio assuming no addition as 100%) in a test group containing 500 μg / mL collagen. [Figure 26] FIG. 1 is a diagram showing the function of water-soluble ionized silicon. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0030] The inventors focused on the effectiveness of lactic acid bacteria fermentation extract derived from brown sugar, soybean pulp, or soybeans, and discovered that by combining plum extract with hydrogen, deep sea water, and other highly functional ingredients, it is possible to obtain significant bactericidal and anti-aging effects, which led to the creation of the present invention.

[0031] In other words, the lactic acid bacteria production substance-containing composition of the present invention is a lactic acid bacteria production substance-containing composition using a lactic acid bacteria fermentation extract consisting of a lactic acid bacteria production substance, and is characterized by containing a lactic acid bacteria fermentation extract derived from brown sugar, soybean pulp, or soybean, and deep sea water, as well as containing 22% or more by weight of plum extract, which is a plant of the Rosaceae family.

[0032] As a result, the present inventors have been able to provide a composition containing a lactic acid bacteria production substance that is capable of exerting a bactericidal effect against so-called "bad bacteria" in the body and an anti-aging effect without using chemical substances (synthetic chemical substances). Hereinafter, embodiments of the present invention will be specifically described.

[0033] In this embodiment, the "composition containing a lactic acid bacteria production substance" according to the present invention is called "reduced fermenting lactic acid bacteria". This reduced fermenting lactic acid bacteria contains a lactic acid bacteria fermentation extract derived from brown sugar, soybean pulp, or soybean. Here, "lactic acid bacteria fermentation extract derived from brown sugar, soybean pulp, or soybean" means a lactic acid bacteria production substance cultured using brown sugar, soybean pulp, or soybean as food for a complex group of lactic acid bacteria. That is, the reduced fermenting lactic acid bacteria according to this embodiment contains "a lactic acid bacteria production substance cultured using brown sugar as food (medium) for a complex group of lactic acid bacteria", "a lactic acid bacteria production substance cultured using soybean pulp as food (medium) for a complex group of lactic acid bacteria", or "a lactic acid bacteria production substance cultured using soybean as food (medium) for a complex group of lactic acid bacteria". The preferred state of this lactic acid bacteria fermentation extract is not particularly limited, and may be liquid, powder, or granules. In addition, the lactic acid bacteria may be live or dead. The reduced fermented lactic acid bacteria according to this embodiment contains deep seawater and 22% or more by weight of plum extract, which is a plant of the Rosaceae family. Plum extract is available in various forms, such as powder, fruit juice, and concentrated fruit juice, but in the present invention, any form is considered to be an original liquid (or a form equivalent to an original liquid).

[0034] Although reduced fermented lactic acid bacteria are acidic, when they are placed in the mouth, the umami taste created by the sourness and sweetness of the bacteria causes saliva to be secreted quickly, neutralizing the acidity. This acts as a training for promoting secretion of salivary glands, and the addition of calcium and phosphate ions contained in saliva is expected to promote remineralization of teeth. This is expected to have an effect of suppressing periodontal disease and bad breath. Furthermore, as described below, the addition of deep seawater minerals that have been reduced and ionized by hydrogen is expected to have a positive effect.

[0035] [Mechanism of reduced fermentation lactic acid bacteria] FIG. 4 is a diagram showing an outline of the mechanism of reduced fermentation lactic acid bacteria. As shown in FIG. 4, reduced fermentation lactic acid bacteria are produced by mixing hydrogen and deep sea water at optimal concentrations during co-culture of multiple types of lactic acid bacteria (e.g., specific 8 or 12 types) and fermenting them to mature. This antibacterial peptide P is secreted by the co-cultured lactic acid bacteria to compete with each other and protect themselves from other bacteria. Then, when this antibacterial peptide P is produced, plum extract is added. This produces an antibacterial peptide with high antibacterial activity (shown as "P+" in FIG. 4). This antibacterial peptide P+ has high antibacterial activity, and by adjusting the concentration, it can be used for oral care and gastrointestinal care. In other words, it is expected to act on the inhibition of bad bacteria present in the body and make the internal environment healthy. Thus, reduced fermentation lactic acid bacteria were developed for the purpose of inhibiting bad bacteria, through research into combining multiple materials to enhance synergistic effects based on the inventors' knowledge of health and basic principles.

[0036] Traditionally, in appealing to the intestinal flora, the main focus has been on increasing good bacteria by ingesting lactic acid bacteria, bifidobacteria, oligosaccharides, etc. In other words, the aim has been to increase the good bacteria, thereby relatively decreasing the ratio of bad bacteria and improving the intestinal environment. However, in addition to increasing the good bacteria, we should pursue suppressing the bad bacteria themselves. From the perspective of "inner beauty", the present inventors have found an approach to reduce bad bacteria in the intestines and oral cavity in addition to an approach to increase good bacteria. Figure 5A is a diagram showing the logic of the present inventors, and Figure 5B is a diagram showing the function of lactic acid bacteria fermentation extract (Biogenics). According to the reduced fermentation lactic acid bacteria of this embodiment, the action of "lactic acid bacteria (live bacteria / dead bacteria)" and "lactic acid bacteria fermentation extract (Biogenics)" increases good bacteria and improves the intestinal environment, and the action of antibacterial peptide P+ is expected to reduce bad bacteria. In other words, it is possible to expect "antioxidation based on the effect of reducing bad bacteria," which is the main cause of active oxygen, which is said to account for 90% of the production in the intestines, and "anti-glycation based on the effect of suppressing the production of AGEs," which cause aging and disease when they accumulate. Based on measures against "oxidation and glycation," the two major causes of aging and disease, and equipped with additional functionality, the reduced fermentation lactic acid bacteria of this embodiment have been realized.

[0037] The lactic acid bacteria fermentation extract contains 377 (mg / L) to 566 (mg / L) peptides. That is, the lactic acid bacteria fermentation extract was filtered through a 0.45 μm filter and used as a measurement sample. The measurement sample was subjected to total peptide quantification using "Pierce Quantitative Colorimetric Peptide Assay (manufactured by Thermo Fisher Scientific)". It was found that the lactic acid bacteria fermentation extract contains 472 (mg / L) peptides. If the effective range is defined as a range of ±20% from this value, it is preferable that the peptides are contained in an amount of 377 (mg / L) to 566 (mg / L). It may also be contained as lactic acid bacteria cells. Since lactic acid bacteria secrete peptides having antibacterial properties to protect themselves, this embodiment uses these peptides as a bactericidal function. In addition, the lactic acid bacteria cells include both live bacteria and dead bacteria. This is because either can provide an effect. In this embodiment, it is preferable that the peptide is contained at 377 (mg / L) to 566 (mg / L), but research by the inventors has shown that a peptide concentration of about 1000 (mg / L) can be realized. Therefore, the present invention is not limited to this numerical range. When actually producing the reduced fermenting lactic acid bacteria according to this embodiment, it is also possible to use a KS melt lactic acid bacteria production substance derived from brown sugar. However, the present invention is not limited to this, and it is also possible to use a lactic acid bacteria production substance derived from soybean pulp or soybeans.

[0038] As for lactic acid bacteria, the following types can be used, for example, within the scope of legal restrictions. In addition, many other types of lactic acid bacteria and bifidobacteria may be added to the complex culture. [Table 1]

[0039] In addition, the reduced fermented lactic acid bacteria (powder) according to the present embodiment can be adjusted to "lactic acid bacteria count: 100 billion / g" by, for example, a composite culture of 12 kinds of lactic acid bacteria. In addition, for example, the reduced fermented lactic acid bacteria (powder) according to the present embodiment can be adjusted to "lactic acid bacteria count: 50 billion / g" by, for example, a composite culture of 8 kinds of lactic acid bacteria. In this way, the reduced fermented lactic acid bacteria (powder) according to the present embodiment contains about "30 to 150 billion / g" of lactic acid bacteria. When calculated based on the number of lactic acid bacteria (1 billion) specified in the fermented milk ingredient standard of the Milk and Milk Products Ordinance, assuming that one commercially available yogurt is 100 mL, by taking "1 g" of the reduced fermented lactic acid bacteria powder, it is possible to take in the amount of lactic acid bacteria equivalent to 30 to 150 commercially available yogurts. Note that these values ​​are examples for practical use, and the present invention is not limited thereto.

[0040] Deep ocean water is generally understood to be seawater at a depth of 200 m or more, and has the characteristics of cleanliness, abundance of inorganic nutrients, and low-temperature stability compared to surface water. In other words, deep ocean water is not affected by river water polluted by human wastewater, so it is not contaminated by chemicals, and since plankton cannot grow because sunlight does not reach it, the amount of harmful bacteria is less than one thousandth of that of surface water. In addition, compared to surface water, it is rich in inorganic nutrients necessary for the growth of phytoplankton, and furthermore, it has the characteristics that the water temperature and contained components are less likely to change, and the water quality is stable. The reduced fermented lactic acid bacteria according to this embodiment contain 0.03 to 5% of deep ocean water reduced by hydrogen. By performing reduction treatment using hydrogen, hydrogen is absorbed, and it is thought that when dissolved in water, it contributes to promoting the ionization of minerals such as calcium and magnesium.

[0041] In addition, in this embodiment, an example is shown in which deep sea water is subjected to reduction treatment with hydrogen and then mixed with lactic acid bacteria fermentation extract, etc., but the present invention is not limited to this, and it is also possible to perform reduction treatment with hydrogen after the oral composition is completed. Note that the present invention does not necessarily require reduction treatment with hydrogen for deep sea water. In addition, deep sea water may be handled in a powdered state for convenience, but the present invention is not limited to powder. In addition, deep sea water in which minerals have been concentrated using ion exchange technology with activated carbon may be added.

[0042] The inventors conducted a test on the ability of "deep ocean water that has been reduced using hydrogen" (sometimes referred to as "ultra-concentrated solution of reduced minerals") to eliminate alcohol and acetaldehyde. The equipment used was a gas chromatograph GC-2010AF / AOC (manufactured by Shimadzu Corporation), and the reagents used were ethanol 95% (manufactured by Wako Pure Chemical Industries, Ltd.), acetaldehyde (manufactured by Wako Pure Chemical Industries, Ltd.), and whiskey (Suntory Kakubin). The test method was as follows. [About alcohol] (a) Three drops of the sample were added to 100 mL of whiskey concentrate (40% alcohol), stirred, and then allowed to stand for two hours, after which the alcohol content was measured. (b) Six drops of the sample were added to 100 mL of whiskey concentrate (40% alcohol), stirred, and then allowed to stand for two hours before measuring the alcohol content. [About acetaldehyde] Dilute acetaldehyde (90%) 100 times to make a 0.9% solution. (A) Three drops of the sample were added to 100 mL of (a) above, stirred, and the acetaldehyde concentration was measured after 30 minutes. (B) Six drops of the sample were added to 100 mL of (a) above, stirred, and the acetaldehyde concentration was measured after 30 minutes. The results are as shown in the following table. [Table 2] In this way, it was confirmed that the "deep ocean water that has been subjected to reduction processing using hydrogen" according to this embodiment exhibits the function of eliminating alcohol and acetaldehyde. Acetaldehyde is known to be a bad substance that causes hangovers and bad hangovers and is involved in "oxidation and glycation of the body." The "deep ocean water that has been subjected to reduction processing using hydrogen (reduced mineral concentrated solution)" according to this embodiment has high acetaldehyde elimination activity, and is therefore expected to suppress oxidation and glycation of the body.

[0043] As described above, by processing deep sea water with hydrogen, it contains essential and trace minerals such as calcium, magnesium, and silicon, and both hydrogen gas and hydrogen ions carrying electrons can be efficiently generated and can be dissolved in water for a long time. Regarding this "electron-carrying hydrogen ion," while general hydrogen ions are "H+" that are missing one electron, there are "negative hydrogen ions H-" that are charged with two electrons, and it has been reported that research toward practical use is being conducted at Kyoto University and Tokyo Institute of Technology. If you drink the "deep sea water that has been reduced with hydrogen" according to this embodiment, you can expect to efficiently deliver hydrogen to the stomach and intestines, liver, and blood. In addition, if you use it as a cosmetic product applied to the skin or scalp, such as a hydrogen gel pack or hydrogen treatment, you can expect to efficiently deliver hydrogen from the surface of the skin or scalp to the endothelium, eliminating the active oxygen that is generated by damage caused by ultraviolet rays.

[0044] Next, we will explain silicon and its related substances. Here, based on the International Atomic Weight Table (2010), the atomic weights are "Si 28.0855", "H 1.00794", and "O 15.9994", rounded off to the third decimal place. Silicon is expressed as "Si" and has an atomic weight of 28.09. The amount of silicon required by the human body per day is "10 to 40 mg", and the recommended intake amount of silicon is used. Next, silica is also called silicic acid, anhydrous silicic acid, and silicon dioxide, and is expressed as "SiO2", with a molecular weight of 60.09 (28.09 + 16.00 × 2 = 60.09). Although it is anhydrous and not water-soluble silicon that is easily absorbed by the body, it is sometimes called "silica" as an alternative name for water-soluble silicon because the word "silica" sounds good. However, water-soluble silicon (hydrated silica / hydrated silicic acid such as metasilicic acid or orthosilicic acid) and silica (anhydrous silicic acid) are different substances, as shown in the molecular formula and molecular weight below. Silica (silicic acid / anhydrous silicic acid) is hydrated and one H2O is added to turn it into metasilicic acid, and it is further hydrated and one H2O is added to turn it into orthosilicic acid, which is easily absorbed by the body and used effectively. Water-soluble silicon is hydrated silica (hydrated silicic acid), a substance that takes the form of metasilicic acid or orthosilicic acid. Insoluble silica (silica / silicic acid / anhydrous silicic acid) that makes up mountains and rocks combines with H2O and dissolves, turning into metasilicic acid. It is further hydrated (adding H2O) and turns into orthosilicic acid, which is more easily absorbed by the body, and reaches the ocean, where it is absorbed by the "phytoplankton diatoms" at the bottom of the food chain, and is used by the body. To repeat, the "drinkable silica derived from hot springs and spring waters" used in mineral water is "water-soluble silicon," which dissolves from rocks (silica, silicic acid, anhydrous silicic acid) and refers to "metasilic acid (hydrated silica, hydrated silicic acid)" or "orthosilicic acid," which have long been used as skin-beautifying bath ingredients in drinking springs. This results in "silicon / silica" = "28.09 / 60.09" = "0.47" times. Also, "silica / silicon" = "60.09 / 28.09" = "2.14" times, so silicic acid (silica, SiO2) = silicon (Si) x 1 / 0.47 = silicon (Si) x 2.14 (based on the conversion value used by the Japan Food Analysis Center).

[0045] Next, metasilicic acid is expressed as "H2SiO3" and has a molecular weight of 78.1 (1 x 2 + 28.09 + 16.00 x 3 = 30.09 + 48.0 = 78.09). From this, it can be said that metasilicic acid is formed when silica (silicic acid, silicon dioxide) is hydrated and changed into "H2O + SiO2 = H2SiO3". In addition, the following relationship is found: "Silicon / Metasilicic acid" = "28.09 / 78.09" = "0.36" times "Metasilicic acid / silicon" = "78.09 / 28.09" = "2.78" times "Silica / Metasilicic Acid" = "60.09 / 78.09" = "0.77" times "Metasilicic acid / silica" = "78.09 / 60.09" = "1.30" times

[0046] Next, orthosilicic acid is expressed as "H4SiO4" and has a molecular weight of 78.1 (1 x 4 + 28.09 + 16.00 x 4 = 32.09 + 64.0 = 96.09). Compared to the molecular formula of metasilicic acid above, it has an additional "H2O" added to make it hydrated. Silica exists mainly in the form of "orthosilicic acid (H4SiO4)" and its biogeochemical cycle is controlled by diatoms. This "orthosilicic acid (H4SiO4)" is "Si(OH)4" and has a "beautiful molecular structure with four OH groups clasping hands" with Si at the center, making it highly absorbent. In addition, the following relationship is found: "Silicon / orthosilicic acid" = "28.09 / 96.09" = "0.29" times "Orthosilicic acid / silicon" = "96.09 / 28.09" = "3.42" times "Silica / orthosilicic acid" = "60.09 / 96.09" = "0.63" times "Orthosilicic acid / silica" = "96.09 / 60.09" = "1.60" times

[0047] Here, we will explain the recommended daily intake. As mentioned above, "silica / silicon" = "60.09 / 28.09" = "2.14" times, "metasilic acid / silicon" = "78.09 / 28.09" = "2.78" times, "orthosilicate / silicon" = "96.09 / 28.09" = "3.42" times. Since the recommended daily intake of silicon alone is "10 to 40 mg", the recommended daily intake when converted to silica, metasilicic acid, and orthosilicate is considered to be within the range of each conversion value from the silicon amount. In other words, the amount of silicon is 10 to 40 mg, the amount of silica is 21.4 to 85.6 mg (10 to 40 mg x 2.14 times), the amount of metasilicic acid is 27.8 to 111.2 mg (10 to 40 mg x 2.78 times), and the amount of orthosilicic acid is 34.2 to 136.8 mg (10 to 40 mg x 3.42 times).

[0048] According to the paper "Biochemistry of Silicon and Related Problems (Nobel Foundation Symposia)" edited by "Gerd Bendz," "the silicon content in the human aorta changes with age." As shown in Figure 16, if the amount of water-soluble silicon (hydrated silica and hydrated silicic acid such as metasilicic acid or orthosilicic acid) in the body at birth is taken as 100, it will decrease to about half by the age of 40, ignoring individual differences. Humans cannot produce the necessary water-soluble silicon in their own bodies, so it is important to actively ingest water-soluble silicon in order to maintain beauty and health.

[0049] Water-soluble silicon (hydrated silica and hydrated silicic acid such as metasilicic acid or orthosilicic acid) is also found in the human body, and is present in hair, nails, blood vessels, bones, joints, cell walls, etc. Silicon in the body has the ability to bind collagen, and is useful for regenerating, strengthening, and maintaining bones, hair, nails, and collagen, as well as affecting skin moisturization. Silicon is also found in the skin (dermis layer), hair, nails, etc., and binds collagen, ceramide, elastin, hyaluronic acid, chondroitin, etc., maintaining the firmness and elasticity of the skin and binding and strengthening tissues. Furthermore, it is known that the combination of ceramide or hyaluronic acid, lactic acid bacteria, and silicon can provide excellent beauty and health effects. That is, ceramide enhances the barrier function of the skin (especially the moisturizing effect) and has the function of suppressing the evaporation of moisture from the epidermis, hyaluronic acid enhances the water retention function of the skin and prevents dryness, lactic acid bacteria have an intestinal regulation effect as well as an immune-boosting function and an anti-allergic function, and silicon promotes the synthesis of collagen in the skin and has the function of bonding and improving the quality of collagen layers, so it is expected that the combination of these will produce beauty and health effects. Furthermore, silicon is absorbed through the intestinal wall and has the effect of solubilizing adhesions inside the blood vessels when passing through the blood vessels, and is also effective in preventing arteriosclerosis. Furthermore, it has the function of promoting plant growth or strengthening stems.

[0050] In this way, since a synergistic effect on beauty can be expected, in addition to the reduced fermentation lactic acid bacteria according to this embodiment, lactic acid bacteria, lactic acid bacteria fermentation extract, oligosaccharides, various polyphenols (plum extract, rose petal extract, pomegranate extract, camellia seed extract, camellia flower extract, etc.), plant enzymes, and citric acid may be added. Collagen, ceramide, elastin, hyaluronic acid, chondroitin, proteoglycan, bird's nest extract, GABA (gamma-aminobutyric acid), and soybean germ isoflavone-derived "equol" may also be added.

[0051] The Framingham Offspring Study in the United States found that there is a close relationship between silicon intake (contained in water-soluble silicon) and bone mineral density (BMD). In this study, 2,846 male and female study subjects in their 30s to 80s were divided into four groups according to the results of their silicon intake and compared. As a result, it was found that the higher the silicon intake, the higher the femoral neck bone density in men and premenopausal women. This suggests that silicon is effective in preventing osteoporosis. Since the importance of silicon has been revealed, dietary supplements and supplements containing water-soluble silicon (hydrated silica and hydrated silicic acid such as metasilicic acid or orthosilicic acid), which is easily absorbed by the body, have been attracting attention in Europe and the United States for quite some time, and the market for silicon products in Europe and the United States is already very large. Human bones are composed of about 70% hydroxyapatite and about 30% collagen, and water-soluble silicon is what runs through them and makes them strong as tissue. For this reason, by including silicon, hydroxyapatite, and collagen, it is expected that the synergistic effect of the collaboration of these three ingredients will be large. In this embodiment, "Sacran" may be added. Sacran is a substance belonging to the "polysaccharides" category, the same as "hyaluronic acid," which is an essential moisturizing agent for cosmetics, and "chondroitin sulfate," which is used in health foods, and is produced from the agar-like substance of Aphanothece sacrum, a freshwater photosynthetic microorganism that is an edible blue-green algae unique to Japan. Sacran is highly viscous and has high moisturizing, anti-inflammatory, and film-forming functions.

[0052] Also, instead of the soybean germ isoflavone-derived equol, a yam extract containing diosgenin may be added. This yam extract containing diosgenin has estrogenic effects, making it an alternative source of equol. Furthermore, it may contain "wild yam extract powder," which is the most nutritious and invigorating of all yams. Coprino and Tamogitake mushrooms, which are rich in the antioxidant amino acid "ergothioneine," may be added, and mushroom extracts such as Himematsutake and Beni-Kusunokitake, which are expected to have excellent immune-stimulating and anti-tumor effects due to their "β-glucan protein complex," may be added. In addition, excipients such as inulin, acacia dietary fiber, soluble dietary fiber such as resistant dextrin, cellulose, and dextrin, starch, and insoluble dietary fiber may be added. Furthermore, it is possible to add excellent antioxidant and anti-glycation raw materials such as red pine bark extract powder, larch bark extract powder, pine bark (pycnogenol), astaxanthin, coenzyme Q10, fullerene, etc., and it is also possible to add raw materials that are expected to have anti-glycation, hair growth, and diet effects such as salacia, fermented soy isoflavones (hydroxylated isoflavones), fucoidan, kudzu flower isoflavones, etc. In addition, it is possible to add SENSEPUR (registered trademark) as a "rice germ and soybean fermented extract" containing yucca extract and quillaja extract to enhance the deodorizing effect.

[0053] In particular, the combination of red pine bark extract powder and astaxanthin is expected to have excellent antioxidant and anti-glycation functions. Furthermore, by blending hydrogen, water-soluble silicon, deep sea water, or adding red or orange coloring, the appeal of the product can be enhanced, and collaboration products with the reduced fermentation lactic acid bacteria of this embodiment are expected to be realized. In addition, collaboration between red pine bark extract powder and Princess Matsutake extract powder or "mushroom beta-glucan" such as Camphor Tree Mushroom is expected to enhance immune activation, and collaboration with Coprino Tamogitake, which is rich in the antioxidant amino acid "ergothioneine," is expected to enhance antioxidant effects.

[0054] The inventors also verified the α-glucosidase inhibitory activity of red pine bark extract powder through a third-party organization. Sugars contained in food are broken down from polysaccharides to monosaccharides by decomposing enzymes and absorbed through the intestinal wall. The enzyme that breaks down disaccharides into monosaccharides at the end of this decomposition process is α-glucosidase. Inhibiting α-glucosidase prevents the breakdown of disaccharides into monosaccharides, suppressing the absorption of sugar from the small intestine. As a result, it is expected to have an effect of suppressing a sudden rise in blood sugar levels after meals. The test method is as follows. That is, the provided sample was dissolved in 50% DMSO at a concentration of 1 mg / mL and filtered through a 0.45 μm filter to prepare the stock solution. The α-glucosidase inhibitory activity was measured for the stock solution and solutions diluted 10-fold and 100-fold with 50% DMSO. The substrate solution was prepared by dissolving "7 mM p-nitrophenyl-α-D-glucopyranoside" in 50 mM phosphate buffer (pH 7.0), and the enzyme solution was prepared by dissolving α-glucosidase at 0.9 U / mL in 50 mM phosphate buffer (pH 7.0). 10 μL of the sample solution and 40 μL of the enzyme solution were placed in a microtube and incubated at 37°C for 5 minutes, after which 950 μL of the substrate solution was placed and incubated at 37°C for 15 minutes. Then, 1000 μL of 0.5 M Tirs solution was added to stop the reaction. The absorbance at 405 nm of p-nitrophenol released through decomposition in this reaction was measured. The inhibition rate of p-nitrophenol production when the sample was added was calculated from the decrease in absorbance from when no sample was added, and this was taken as the α-glucosidase inhibitory activity. The results are shown in the table below, and indicate that the higher the α-glucosidase inhibitory activity, the greater the effect of suppressing the rise in blood glucose level. [Table 3] Thus, the red pine bark extract powder of this embodiment was found to have α-glucosidase inhibitory activity in both the undiluted solution, 10-fold dilution, and 100-fold dilution, and particularly high α-glucosidase inhibitory activity was confirmed in the undiluted solution.

[0055] The inventors also verified the Amadori compound production inhibitory activity of red pine bark extract powder through a third-party organization. Amadori compounds are important intermediates in the early stage of glycation reactions, and by inhibiting the production of Amadori compounds, it is possible to inhibit the production of AGEs, which are late products of glycation reactions. The test method is as follows. That is, the provided sample was dissolved in 50% DMSO at a concentration of 1 mg / mL and filtered through a 0.45 μm filter to prepare a stock solution. The stock solution and solutions diluted 10-fold and 100-fold with 50% DMSO were used as measurement samples to measure the Amadori compound production inhibitory activity. For the measurement, the measurement sample, glucose solution, and bovine serum albumin solution were mixed and incubated at 60°C for 48 hours to prepare a glycation reaction solution. 300 μL of 0.25 mM nitroblue tetrazolium solution (pH 9.0) was added to 40 μL of the glycation reaction solution, incubated at 37°C for 30 minutes, and then allowed to stand at room temperature for 16 hours, after which the absorbance at 540 nm was measured. The inhibitory activity against the formation of Amadori compounds when the sample was added was calculated from the change in absorbance, and this was taken as the inhibitory activity against the formation of Amadori compounds. The results are shown in the following table, with higher inhibitory activity indicating higher anti-glycation activity. [Table 4] Thus, the red pine bark extract powder according to this embodiment was found to have inhibitory activity against the production of Amadori compounds in the undiluted solution and in a 10-fold dilution.

[0056] The inventors also verified the AGEs cross-linking cleavage activity of red pine bark extract powder through a third-party organization. α-Dicarbonyl compounds are known to be intermediates produced in the glycation process of proteins in vivo, and include 3-deoxyglucosone and glyoxal. AGEs, which are produced by non-enzymatic reaction (Maillard reaction) between α-dicarbonyl compounds produced in vivo and amino groups of proteins, are the cause of various diseases, as mentioned above. It is expected that the glycation of proteins will be prevented by ingredients that have the activity of decomposing α-dicarbonyl compounds, i.e., the activity of decomposing AGEs cross-linking.

[0057] The AGEs cross-link cleavage test was performed as follows. That is, the amount of benzoic acid generated by decomposition of the dicarbonyl bond of the model compound "1-phenyl-1,2-propanedione" having a dicarbonyl bond was measured. The provided sample was dissolved in 50% DMSO aqueous solution at a concentration of 1 mg / mL and filtered through a 0.45 μm filter to prepare the stock solution. The stock solution and the solution diluted 10-fold and 100-fold with 50% DMSO aqueous solution were used as the measurement samples. The measurement sample, "1-phenyl-1,2-propanedione solution," and methanol were mixed to prepare the reaction solution. After incubating the reaction solution at 37°C for 24 hours, hydrochloric acid was added to stop the reaction. The amount of benzoic acid generated by decomposition in the reaction solution was quantified using high-performance liquid chromatography. The AGEs cross-link cleavage rate was calculated as 100 (%) when all of the "1-phenyl-1,2-propanedione" in the reaction solution was decomposed to benzoic acid. The results are as shown in the following table. [Table 5] Thus, the red pine bark extract powder according to this embodiment was found to be able to cleave AGEs cross-links in the undiluted solution and at a 10-fold dilution. Note that the "-" in the 100-fold dilution indicates that the AGEs cross-link cleavage rate could not be calculated because the concentration of benzoic acid was below the quantification limit.

[0058] The present inventors verified the "fluorescent AGEs production inhibitory activity" and "non-fluorescent AGEs (CML) production inhibitory activity" of red pine bark extract powder through a third-party organization. AGEs (Advanced Glycation End products) are substances that are generated when reducing sugars such as glucose react non-enzymatically with amino groups in proteins (Maillard reaction). When AGEs accumulate in blood vessels, they cause myocardial infarction and cerebral infarction, and when they accumulate in bones, they cause osteoporosis, and when they accumulate in the eyes, they cause cataracts. AGEs are broadly divided into fluorescent AGEs that have fluorescence and non-fluorescent AGEs that do not have fluorescence.

[0059] The fluorescent AGEs formation inhibitory activity test method is as follows. The provided sample was dissolved in 50% DMSO at a concentration of 1 mg / mL and filtered through a 0.45 μm filter to prepare a stock solution. The fluorescent AGEs formation inhibitory activity was measured for the stock solution and solutions diluted 10-fold and 100-fold with 50% DMSO. Glucose and bovine serum albumin were mixed in a test tube and incubated at 60°C for 40 hours, after which the generated fluorescent AGEs were measured using a fluorescence spectrophotometer at an excitation wavelength of 370 nm and a fluorescence wavelength of 440 nm. The rate of inhibition of fluorescent AGEs formation when the sample was added was calculated from the decrease in fluorescence intensity, and this was taken as the fluorescent AGEs formation inhibitory activity. The results are shown in the following table, with a higher inhibition rate indicating a higher anti-glycation activity. [Table 6] Thus, it was confirmed that red pine bark extract powder has the effect of inhibiting the production of fluorescent AGEs.

[0060] Next, we will explain the test method for the inhibitory activity against the formation of non-fluorescent AGEs (CML). CML (carboxymethyllysine) is a representative non-fluorescent AGE, and is generated by glycation modification of lysine residues of proteins. It is expected that the components that have the inhibitory effect against the formation of CML will prevent the glycation of proteins. The test method is as follows. That is, the sample was dissolved in 50% DMSO aqueous solution at a concentration of 1 mg / mL and filtered through a 0.45 μm filter to prepare the stock solution, and the stock solution and a solution diluted 10 times and 100 times with 50% DMSO aqueous solution were used as the measurement samples. The measurement sample, glucose, and bovine serum albumin were mixed in a microtube and incubated at 60°C for 40 hours, after which a portion was taken, a protease solution was added, and the mixture was incubated at 37°C for 24 hours to perform enzymatic degradation. After that, substances with a molecular weight of 10,000 or more were removed by ultrafiltration, and CML was analyzed by liquid chromatography mass spectrometry. The rate of inhibition of CML formation when the sample was added was calculated from the change in CML concentration, and this was taken as the inhibitory activity against non-fluorescent AGEs (CML) formation. The higher the inhibitory activity, the greater the inhibitory effect against non-fluorescent AGE formation. The results are as follows: [Table 7] In this way, it was shown that red pine bark extract powder has a high inhibitory effect on the production of non-fluorescent AGEs. In addition, "rose petal powder" is known to have anti-glycation, skin-beautifying, and diet effects. For this reason, by combining red pine bark extract powder with rose petal powder, a synergistic effect on anti-glycation, skin-beautifying, and dieting is expected.

[0061] The inventors also verified the hyaluronidase inhibitory activity of red pine bark extract powder through a third-party organization. Hyaluronic acid exists around cells and between cells in the body, and maintains the moisture of the skin to maintain healthy skin and is involved in the transport of nutrients and waste products. The amount of hyaluronic acid in the body decreases with age, causing arthritis, wrinkle formation, etc. On the other hand, hyaluronidase is an enzyme that breaks down hyaluronic acid and is widely present in the body. It is thought that it is activated during inflammation, destroys the structure of tissues, and promotes inflammation. Inhibiting hyaluronidase can prevent the decomposition of hyaluronic acid, and is expected to have a moisture retention function effect on the skin and an inflammation suppression effect.

[0062] The test method for hyaluronidase inhibitory activity is as follows. The provided sample was dissolved in 50% DMSO aqueous solution at a concentration of 1 mg / mL and filtered through a 0.45 μm filter to prepare the stock solution. The stock solution and the solution diluted 10-fold and 100-fold with 50% DMSO aqueous solution were used to measure the hyaluronidase inhibitory activity. The enzyme solution was prepared by dissolving bovine hyaluronidase in 0.1 M acetate buffer (pH 4.0) at 400 units / mL. The substrate solution was prepared by dissolving potassium hyaluronate in acetate buffer at 1 mg / mL. The enzyme activator was prepared by dissolving Compund48 / 80 in acetate buffer at 0.1 mg / mL. The color developing solution was prepared by dissolving p-dimethylaminobenzaldehyde (p-DABA) in a hydrochloric acid / acetic acid mixture at 100 mg / mL, and was used after diluting 10-fold with acetic acid just before use.

[0063] The measurement was carried out by adding 12 μL of sample solution and 12 μL of enzyme solution to a microtube and incubating at 40°C for 20 minutes. 12 μL of enzyme activator was added thereto and incubated at 40°C for 20 minutes. 12 μL of substrate solution was added and incubated at 40°C for 40 minutes. 12 μL of 0.4N NaOH aqueous solution was then added to stop the reaction, and the mixture was immediately cooled on ice for 5 minutes, 12 μL of 0.8M borate buffer (pH 9.0) was added, boiled for 3 minutes, and then cooled on ice for 10 minutes. 180 μL of color developing solution was added thereto and incubated at 40°C for 30 minutes, after which the absorbance at 585 nm was measured. If the activity of hyaluronidase is inhibited when the sample is added, N-acetylglucosamine, a decomposition product of hyaluronic acid, decreases, and the absorbance of p-DABA decreases. The amount of change in absorbance was calculated to determine the hyaluronidase inhibitory activity. The results are as shown in the following table. [Table 8] Thus, the red pine bark extract powder according to this embodiment was found to have hyaluronidase inhibitory activity in all cases, whether it was the undiluted solution, a 10-fold dilution, or a 100-fold dilution.

[0064] The inventors also verified the collagenase inhibitory activity of red pine bark extract powder through a third-party organization. Collagen accounts for about 1 / 3 of the total protein in the body, is found in skin and bones, and is greatly involved in maintaining their structure. Collagenase is an enzyme that breaks down collagen, and when collagenase is activated by aging or ultraviolet rays, collagen is broken down, making it difficult to maintain the structure of the skin, leading to the occurrence of wrinkles and sagging. By adding a collagenase inhibitor to cosmetics, anti-aging effects against wrinkles and sagging can be expected.

[0065] The test method is as follows. The provided sample was dissolved in 50% DMSO aqueous solution at a concentration of 1 mg / mL, and filtered through a 0.45 μm filter to prepare the stock solution. Collagenase inhibitory activity was measured for the stock solution and solutions diluted 10-fold and 100-fold with 50% DMSO aqueous solution. Collagenase was dissolved in pure water at 0.1 mg / mL to prepare the enzyme solution, and Pz-peptide was dissolved in 0.1 M Tris buffer (pH 7.1, 20 mM CaCl2) at 0.39 mg / mL to prepare the substrate solution. For the measurement, 12.5 μL of sample solution, 12.5 μL of enzyme solution, and 100 μL of substrate solution were placed in a glass test tube, incubated at 37°C for 30 minutes, and then 250 μL of 25 mM citric acid was added to stop the reaction. In this reaction, 1250 μL of ethyl acetate was added to the collagenase-induced degradation products, which were then extracted by shaking, centrifuged at 3000 rpm for 10 minutes, and the upper layer (ethyl acetate layer) was collected and its absorbance at 320 nm was measured. The inhibition rate of Pz-peptide degradation products produced when the sample was added was calculated from the decrease in absorbance from the value when no sample was added, and this was taken as collagenase inhibitory activity. The results are shown in the table below. [Table 9] Thus, the red pine bark extract powder according to this embodiment was found to have high collagenase inhibitory activity in all cases, whether it was the undiluted solution, a 10-fold dilution, or a 100-fold dilution.

[0066] The inventors also verified the elastase inhibitory activity of red pine bark extract powder through a third-party organization. Elastase is an enzyme that breaks down elastin, an elastic fiber that, together with collagen, determines the mechanical properties of connective tissue. When elastase is activated by aging or ultraviolet rays, elastin is broken down, making it difficult to maintain the structure of the skin, leading to the development of wrinkles and sagging. By adding an ingredient that inhibits elastase to cosmetics, it is expected that it will have an anti-aging effect against wrinkles, sagging, and the like.

[0067] The test method is as follows. That is, the provided sample was dissolved in 50% DMSO aqueous solution at a concentration of 1 mg / mL, and filtered through a 0.45 μm filter to prepare the stock solution. The elastase inhibitory activity was measured for the stock solution and the solution diluted 10-fold and 100-fold with 50% DMSO aqueous solution. STANA (N-Succinyl-Ala-AlaAla-p-nitroanilide) was dissolved in Tris buffer (pH 7.4) at 1 mM to prepare the substrate solution, and porcine pancreatic elastase was dissolved in Tris buffer (pH 7.4) at 0.5 units / mL to prepare the enzyme solution. For the measurement, 50 μL of the sample solution, 50 μL of the enzyme solution, and 100 μL of the substrate solution were placed in a microtube and incubated at 37°C for 30 minutes. In this reaction, nitroaniline, which is a substrate decomposition product by elastase, was measured at 405 nm absorbance. The inhibition rate of nitroaniline formation when the sample was added was calculated from the decrease in absorbance from the value when no sample was added, and this was taken as the elastase inhibitory activity. The results are shown in the following table. [Table 10] Thus, the red pine bark extract powder according to this embodiment was found to have elastase inhibitory activity in all cases, whether it was the undiluted solution, a 10-fold dilution, or a 100-fold dilution.

[0068] Both plant-derived silicon and mineral-derived silicon can be processed under the same conditions, such as temperature and pressure, to obtain water-soluble silicon (hydrated silica or hydrated silicic acid, such as metasilicic acid or orthosilicic acid). However, the balance of minerals other than silicon contained in each is different, so if you want to enjoy both the blessings of the earth and the blessings of plants, you can expect a synergistic effect by mixing concentrated solutions of plant-derived water-soluble silicon and mineral-derived water-soluble silicon. In addition, the "water-soluble silicon" can be either "plant-derived silicon" derived from bamboo, sugarcane, Japanese bamboo grass, Japanese silver grass, rice husks, and rice straw, which are all grasses like bamboo, or "mineral-derived silicon" such as crystal and quartz. Water-soluble silicon derived from natural water or hot spring water from Kirishima (Kirishima Mountain Range) or Hakone can also be used. In addition, by utilizing the water-soluble silicon eluted from the Mt. Fuji lava marimo powder, or by combining it with the water-soluble silicon abundant in the spring water and hot spring water of Mt. Fuji, the "blessings of minerals from Mt. Fuji" and the "blessings of minerals from deep sea water" can be expected to have a further synergistic effect of minerals. It can be said that these are the blessings of high and low minerals, and the blessings of a balance of yin and yang minerals. In addition, the spring water, hot spring water, lava, etc. of Mt. Fuji contain "vanadium," which is known to be effective against diabetes. It is known that the rain and snow that fell on the land of Mt. Fuji becomes natural water that incorporates a balanced amount of abundant minerals, including vanadium, over a long period of time as it permeates through the thick basalt layer. In this embodiment, too, the inclusion of "vanadium" can be expected to be effective against diabetes.

[0069] Mineral waters rich in water-soluble silicon are mainly found in the Fuji and Hakone regions and Kyushu. Kyushu has some of the world's leading volcanoes and hot springs, such as Aso, Unzen, Kirishima, Kuju, Sakurajima, and Beppu. The strata in this area contain a lot of silica (silicic acid and anhydrous silicic acid), which is known to dissolve into the water over a long period of time (changing into hydrated silica and hydrated silicic acid, such as metasilicic acid or orthosilicic acid). Regarding the hot spring water of Kirishima, which is rich in water-soluble silicon, there is a myth that "long ago, Izanagi-no-Mikoto and Izanami-no-Mikoto took Hiruko-no-Mikoto, who could not stand, on a boat and had him undergo hot spring treatment at the "Nageki-no-Mori" (Forest of Lamentations) where they arrived." In this embodiment, water-soluble silicon (hydrated silica and hydrated silicic acid, such as metasilicic acid or orthosilicic acid) derived from spring water or natural water of Kirishima or Sakurajima may also be contained.

[0070] Furthermore, mineral water from Sakurajima may contain natural germanium. Germanium binds with oxygen, performs the function of transporting oxygen, and has been found to activate interferon in the body. For this reason, in this embodiment as well, it may be possible to obtain the benefits of natural germanium by using mineral water from Sakurajima. Interestingly, carbon (atomic number 6), silicon (atomic number 14), and germanium (atomic number 32) belong to the same Group 14 element in the periodic table, and have "s" in their valence electrons. 2 p 2 " each has a four-electron electronic structure, and each has unique characteristics that may provide health benefits.

[0071] Thus, it is meaningful to add collagen, ceramide, and silicon to reduced fermentation lactic acid bacteria, and the present inventors verified the synergistic effect on collagen production based on the results of research conducted by a research institute. The test substances were (a) collagen, (b) ultra-concentrated water-soluble ionized silicon, and (c) ceramide (1% ceramide solution), and the test method was as follows. Specifically, normal human skin fibroblasts were cultured and brought into the logarithmic growth phase, while securing the required number of cells, and the normal human skin fibroblasts were plated in a 96-well microplate at 1 x 10 4 The cells were seeded at a concentration of "cells / 100μL / well". Then, pre-culture was performed for 24 hours, the culture medium was removed, and the adjusted test sample solution was added to each well and cultured for 48 hours. The culture supernatant was then collected and measured using an ELISA collagen kit, with the values ​​corrected for cellular protein being shown. The results are shown in the following table. [Table 11]

[0072] Thus, the combination of "collagen 31.25 (μg / mL)" and "ceramide (1% ceramide solution) 125 (μg / mL)" resulted in a proliferation rate of 128.8% (28.8% increase), while the combination of "collagen 31.25 (μg / mL)", "ceramide (1% ceramide solution) 125 (μg / mL)" and "water-soluble silicon 125 (μg / mL)" resulted in a proliferation rate of 148.3% (48.3% increase). The results are shown in Figure 17. In addition, the combination of "collagen 125 (μg / mL)" and "ceramide (1% ceramide solution) 125 (μg / mL)" resulted in a proliferation rate of 165.2% (a 65.2% increase), and the combination of "collagen 125 (μg / mL)", "ceramide (1% ceramide solution) 125 (μg / mL)", and "water-soluble silicon 125 (μg / mL)" resulted in a proliferation rate of 208.8% (a 108.8% increase). These results are shown in Figure 18. Thus, the addition of "water-soluble silicon" was confirmed to have a statistically significant high synergistic effect compared to the collagen-ceramide two-type mixture (significance test (Student's t-test) vs. two-type mixture: p value < 0.001).

[0073] The inventors also verified the synergistic effect on hyaluronic acid production based on the results of research conducted by research institutes. That is, a hyaluronic acid production effect test was conducted using normal human dermal fibroblasts for the test substances "collagen," "silicon RICH ultra-concentrated solution," and "ceramide (1% ceramide solution)," both individually and in combination with each other. The test method was as follows. (A) Normal human dermal fibroblasts were cultured to ensure the required number of cells. (B) Normal human dermal fibroblasts were cultured in a "96-well microplate" at 1 x 10 4 The cells were seeded onto each plate at a concentration of "cells / 100μL / well". (C) Pre-culture was carried out for 24 hours. (D) After pre-culture, the culture medium was removed and the prepared test substance solution was added to each well, followed by culturing for 48 hours. (E) After completion of the culture, the culture supernatant was collected and the amount of hyaluronic acid produced in the culture supernatant was measured using an ELISA hyaluronic acid kit.

[0074] The calculation method for the test results is as follows. (A) The amount of hyaluronic acid produced was divided by the amount of protein, which is proportional to the number of cells, to calculate the amount of hyaluronic acid produced per protein amount. (B) For relative evaluation, the amount of hyaluronic acid produced in the control group was converted to 100%, and the hyaluronic acid production rate of the test substance was calculated. (C) The significant differences in the cell viability, amount of hyaluronic acid produced, and hyaluronic acid production rate were tested using the Student's t-test with the control. The data for each test group was tested three times independently, and a significant difference test was performed. The significance level was set at p<0.05 in a two-sided test. The test results for the hyaluronic acid production effect of the test substances alone and in combination are shown in the table below. [Table 12] The table above shows the hyaluronic acid production rate results for the test substances alone, and shows the relative ratios with no addition taken as 100%. [Table 13] The table above shows the hyaluronic acid production rate results for the combination of two products, with the relative ratio shown assuming no additives as 100%. [Table 14] The table above shows the hyaluronic acid production rate results for combinations of three products, with the relative ratio shown assuming no additives as 100%.

[0075] For the test substances alone, each test substance was found to have a hyaluronic acid production promoting effect, and collagen and ceramide alone showed high hyaluronic acid production, and a significant production promoting effect was observed in all test groups compared to no addition. For water-soluble silicon, a significant production promoting effect was observed in the 125μg / mL test group compared to no addition. In addition, for the combination of two products, the combination of collagen and water-soluble silicon significantly promoted hyaluronic acid production in all test groups compared to collagen alone, and for the combination of ceramide and water-soluble silicon, the combination with water-soluble silicon promoted hyaluronic acid production in the concentration group of ceramide at 31.25μg / mL or more, and a hyaluronic acid production promoting effect was observed compared to ceramide alone. For the combination of collagen and ceramide, the combination of collagen at 31.25μg / mL or more promoted hyaluronic acid production compared to collagen alone, and a synergistic effect was observed by the combination.

[0076] Furthermore, in a three-item mixture in which water-soluble silicon was added to a combination of collagen and ceramide, the amount of hyaluronic acid produced was significantly promoted in the test group with collagen of 31.25 μg / mL or more compared to the combination of two other items, and the synergistic effect of water-soluble silicon was observed. This is shown in Figures 25A to 25C.

[0077] Silicon is also known to have a significant effect on the growth and shell formation of the Sakura shrimp, known as the jewel of the sea. Sakura shrimp, which are precious worldwide, can only be caught in two places: Taiwan and Suruga Bay. Almost 100% of the shrimp caught in Japan comes from Suruga Bay, Shizuoka Prefecture. As shown in Figure 14, rivers such as the Fuji spring water that flows into Suruga Bay are rich in water-soluble silicon (hydrated silica and hydrated silicic acid such as metasilicic acid or orthosilicic acid), which is an important factor in the formation of the skeleton of phytoplankton called diatoms, which serve as food for Sakura shrimp in their juvenile stage. The diatoms taken in by Sakura shrimp serve as "in vivo silicon" and play a role as a mineral of beauty that is essential for the growth of living organisms through the food chain. That is, silicon travels from water or hot springs through rivers to the sea (Suruga Bay), where it becomes an important component of the body of the phytoplankton "diatoms," and through the rich ecosystem of Suruga Bay, including shrimp, and the food chain, it plays a role in strengthening the skeleton and tissues of the Japanese body. The flow of "transitioning from mineral silicon to plant silicon to biological silicon, passing through the food chain, becoming feces or corpses again, returning to the seabed or the earth, and changing into mineral silicon over a long period of time" is the "silicon circulation on earth." In this way, silicon circulates around the earth over a long period of time, moving between minerals, plants, and living organisms, and can be said to be a "beauty mineral" that is essential to the human body. The composition containing lactic acid bacteria production substances according to this embodiment effectively utilizes the advantages of silicon.

[0078] The present inventors conducted a skin permeability test of the water-soluble silicon according to this embodiment through a third-party organization. In this test, the skin permeability was evaluated by comparing the water-soluble silicon according to this embodiment with commercially available silicon water. That is, the test specimen was the water-soluble silicon according to this embodiment and commercially available silicon water, the subject was a 51-year-old man, and the test site was the inside of the right forearm. In the test method, a predetermined amount of fluorescent agent was added to each test specimen to make the permeability of the test specimen visible. The subject was exposed to the test site and allowed to acclimate for about 10 minutes in a seated position to get used to the environment in the test room under constant conditions (room temperature: 25°C, humidity 50%). After acclimatization, two test sections of 3 cm x 3 cm were set on the test site, one of which was the test section for "water-soluble silicon according to this embodiment" and the other was the test section for "commercially available silicon water". 20 μL of the specimen was dropped into each test section and spread evenly within the test section. Next, distilled water was sprayed over the entire surface of both test areas, and then immediately irradiated with a black light to observe whether fluorescent light was emitted or not.

[0079] As a result, in the test section of "water-soluble silicon according to this embodiment", no coloring of the fluorescent agent was observed, and only the test section of "commercially available silicon water" was observed to have fluorescent coloring. From this, it is considered that the "commercially available silicon water" did not have a skin penetration phenomenon because the fluorescent agent remained on the skin, while in the case of "water-soluble silicon according to this embodiment", it is considered that the fluorescent agent did not remain and have coloring, so it is considered that the fluorescent agent penetrated into the skin. Figure 26 is a diagram showing the function of water-soluble ionized silicon. As shown in Figure 26, silicon binds with collagen and the like, and plays a role in filling gaps in the skin and maintaining elasticity. However, since silicon itself does not easily penetrate the skin, in this embodiment, a "water-soluble ionized silicon" with excellent penetration rate is realized by special processing. This increases the penetration rate, and further effects such as restoring skin vitality and enabling skin reconstruction can be expected.

[0080] It may also contain eggshell membrane peptides or nacre pearl powder. Eggshell membrane contains proteins such as type I collagen, elastin, and keratin, glycosaminoglycans such as hyaluronic acid, chondroitin sulfate, and keratan sulfate, and hexosamines such as glucosamine and hexose, and is known to have anti-inflammatory, analgesic, and cartilage-protecting functions.

[0081] In addition, nacreous pearl powder is composed of multiple types of amino acids, contains conchiolin, which is said to be a beauty ingredient, and contains minerals such as calcium, and is known to have the function of reducing visceral fat and replenishing minerals. This "nacreous layer" is a pearl-colored ceramic layer that covers the inside of the shell, and is secreted from the entire surface of the mantle of the shell. In particular, beautiful nacre is found inside the pearl oyster and black lipped oyster, and it is known that this is reflected in the color of the pearl. It is known that pearls are produced when the shellfish wraps up foreign bodies that it has taken in and cannot expel with its own secretions to protect the body. Because of this process of production, which "transforms suffering into beautiful jewels," pearls are said to "represent the dignity of beautiful women who have overcome difficulties," and there are anecdotes that Yang Guifei and Cleopatra drank pearls, and they have been loved by people all over the world as a symbol of health, longevity, whitening, and wealth. By containing the above "eggshell membrane peptides" or "nacreous pearl powder," beauty effects such as beautiful skin, nails, and hair are expected.

[0082] Keratin may also be included on its own. Keratin is a protein found in large amounts in nails and hair, giving them suppleness and strength. Keratin is made up of a combination of amino acids that is almost the same as the combination of amino acids (called amino acid composition) found in skin, particularly the epidermis, as well as nails and hair. By including keratin, beauty effects such as beautiful skin, nails and hair can be expected.

[0083] The reduced fermentation lactic acid bacteria according to the present embodiment may also contain soy isoflavones including equol, daidzin, or daidzein. In this case, soy isoflavones including daidzin or daidzein may be added to the reduced fermentation lactic acid bacteria according to the present embodiment under a predetermined condition. When soy isoflavone glycosides are ingested, the sugar portion of the soy isoflavone glycoside is separated by the action of the enzymes of the intestinal bacteria in the large intestine, and three types of substances that are not bound to sugar, that is, daidzein, genistein, and glycitein, are generated. These are collectively called soy isoflavone aglycones (sometimes called "aglycone-type isoflavones"). Of these, daidzein is converted to equol by the influence of the intestinal bacteria. When women reach menopause, the secretion of female hormones decreases, and the risk of menopausal disorders and osteoporosis increases. However, equol acts similarly to "estrogen," a female hormone that is directly linked to women's beauty, and is known to be effective against osteoporosis and menopausal disorders. It is also known to be effective in treating men's prostate enlargement and AGA (hair loss). Research has also shown that equol reduces skin wrinkles and inhibits the onset of cancer. Equol is produced by intestinal bacteria, but not everyone has this bacteria in their intestines; it is said that only 20-30% of Westerners and about 40-50% of Japanese people have it. However, the intestinal environment is not uniform or genetically determined, but changes depending on eating habits, etc. Even if a person does not have equol at a certain point, there is a possibility that they will have it later.

[0084] As an example, it has been reported that when Westerners were given aglycone-type isoflavones (soybean isoflavone aglycones) containing daidzein, the equol production rate increased by about 70%. It has also been reported that drinking aglycone-type isoflavones containing a lot of daidzein for three weeks increased the amount of equol produced more than drinking glycoside isoflavones. Furthermore, in a large-scale human clinical trial, it has been reported that drinking aglycone-type isoflavones containing a lot of daidzein increased the blood equol concentration in a dose-dependent manner. The reduced fermented lactic acid bacteria powder according to this embodiment contains equol or daidzein, which improves the intestinal environment, maintains and improves health, and provides an anti-aging effect. In addition, it may contain "soybean extract fermentation product containing hydroxylated isoflavones (fermented isoflavones)" which has been published for its inhibitory effect on the production of AGEs, which is glycation in the body, and its inhibitory effect on glycation hair loss. Lactobionic acid, a functional sugar produced from lactose, promotes calcium absorption and equol production, and is expected to be an anti-aging material that can prevent and alleviate osteoporosis and lifestyle-related diseases. By incorporating lactobionic acid, a synergistic effect with equol is expected. In addition to lactobionic acid produced from lactose, maltobionic acid, which is the same oligosaccharide acid and has similar characteristics to lactobionic acid, such as promoting calcium absorption, has the advantage of being a non-allergenic substance derived from corn starch. The inclusion of maltobionic acid is expected to be effective in promoting equol production.

[0085] The reduced fermented lactic acid bacteria according to this embodiment further contains at least one or more of the following by weight: Rosaceae plant extract, hibiscus extract, cranberry extract, pomegranate extract, olive extract (leaf, fruit, flower), camellia extract (leaf, fruit, flower) or perilla fruit extract. By further mixing various polyphenols such as mangosteen extract, bay leaf extract, quercetin, and eucalyptus, the taste becomes good, it becomes effective for oral care, and the functionality is improved, so that the appeal of the product can be increased.

[0086] Tea seed extract may also be included. Tea seed extract inhibits the α-glucosidase activity required for the absorption of sugar and starch in the digestive tract, thereby inhibiting the absorption of starch and especially sugar. This function reduces the energy intake from carbohydrates, making it possible to balance the calories taken in and the calories burned. By including tea seed extract, it is possible to sufficiently take in nutrients necessary for the human body, such as proteins, vitamins, and minerals, without excessive dietary restrictions, and it is also possible to reduce only the calories taken in from sugar. Furthermore, by taking tea seed extract during meals, it is possible to lower postprandial blood glucose levels.

[0087] It is also possible to add soybean fiber enzymatic hydrolyzate. This soybean fiber enzymatic hydrolyzate is made by removing "protein and lipid" from okara, a by-product of tofu manufacturing, and then partially hydrolyzing the cellulose with cellulase treatment to produce a powder with soluble dietary fiber and insoluble dietary fiber in a ratio of, for example, 1:2. This soybean fiber enzymatic hydrolyzate is used as a material to prevent constipation. Furthermore, by adding tea seed extract to this powder, it is possible to give it the function of inhibiting sugar absorption in addition to preventing constipation.

[0088] In addition, "charcoal or activated carbon with high adsorption performance carbonized by utilizing pressure, reduced water, etc. in addition to temperature (hereinafter referred to as "functional edible carbon (registered trademark)")" may be blended. "Functional edible carbon" adsorbs food additives, heavy metals, odorous substances such as indole and skatole produced by bad bacteria, and "AGEs" (advanced glycation end products) substances formed when proteins and sugars contained in food are heated, and it is possible to reduce bad substances that cause oxidation and saccharification, and to expect a synergistic effect with reduced fermentation lactic acid bacteria. The "functional coconut shell activated carbon" according to this embodiment meets the medicinal charcoal standards and has passed the test of the Japan Food Analysis Center.

[0089] The present inventors conducted an indole and skatole adsorption test for "functional coconut shell activated carbon powder (functional coconut shell activated carbon / palm tree shape is TM)" and "Ina Akamatsu Myotan (registered trademark) powder". The analytical samples were "functional coconut shell activated carbon powder" and "Ina Akamatsu Myotan powder", the equipment used was "gas chromatograph mass spectrometer (Shimadzu Corporation: QP5050A)", and the reagents used were "indole reagent (Wako Pure Chemical Industries, Ltd.)" and "skatole first-class reagent (Wako Pure Chemical Industries, Ltd.)". The test method is as follows. The above indole and skatole are dissolved in water and adjusted to 20 ppm each. Next, 1 g of the reagent is added to 100 mL of the 20 ppm solution, stirred for 10 minutes, and then filtered, and analyzed by the gas chromatograph mass spectrometer. The results are as shown in the following table. [Table 15]

[0090] The inventors also conducted an indole and skatole adsorption test on "Kishu Bincho Activated Carbon Powder" and "Kamakura Keishitake Charcoal (registered trademark) Powder." The samples analyzed were "Kishu Bincho Activated Carbon Powder" and "Kamakura Keishitake Charcoal Powder," the equipment used was a "gas chromatograph mass spectrometer (Shimadzu Corporation: QP5050A)," and the reagents used were "indole reagent (Wako Pure Chemical Industries, Ltd.)" and "skatole reagent (Wako Pure Chemical Industries, Ltd.)." The test method was as follows. The indole and skatole were dissolved in water and adjusted to 20 ppm each. Next, 1 g of the reagent was added to 100 mL of the 20 ppm solution, stirred for 10 minutes, and then filtered, and analyzed by the gas chromatograph mass spectrometer. The results are shown in the following table. [Table 16] As the above two test results show, reduced fermented lactic acid bacteria act to reduce Clostridium perfringens, which is the source of indole, and functional edible charcoal adsorbs indole itself, so intestinal environment care can be expected with a double effect. Skatole is also said to be a foul-smelling gas in tongue coating, which is one of the major causes of bad breath. Functional edible charcoal adsorbs foul-smelling gases such as skatole and methyl mercaptan, and reduced fermented lactic acid bacteria eliminate "bad bacteria in the oral cavity, which are also the source of foul-smelling gases," so oral care can be expected with a double effect.

[0091] Here, the inventors conducted a test to prove that functional edible charcoal has a low abrasive effect on teeth because it is characterized by being carbonized and crushed to a "soft and fluffy" shape using reduced water in addition to temperature and pressure. Since the hardness of the glass surface is weaker than that of tooth enamel, an abrasive effect test was conducted on the glass surface using "Ina Akamatsu Myo Charcoal / Coconut Shell Activated Carbon Granules (Dispersion Processing)" with an electric toothbrush. As a result, it was found that the glass surface was not scratched at all despite being brushed with an electric toothbrush using charcoal for an hour. This shows that the abrasive effect of functional edible charcoal on teeth is completely problem-free. In this embodiment, an example of carbonization and crushing using reduced water is shown, but the present invention is not limited to this. Carbonization and crushing may be performed using reduced water, particularly deep sea water that has been reduced using hydrogen, or water containing water-soluble silicon.

[0092] The effects and efficacy of activated charcoal are recognized within the EU. Specifically, in the category of "reducing excess intestinal gas accumulation," the "EU Health Claim (a statement suggesting a positive effect on health)" states that "activated charcoal contributes to reducing excessive abdominal bloating after meals," and the "labeling conditions" state that "it can only be used for foods that contain 1g of activated charcoal per serving." It is also permitted to "provide information to consumers that beneficial effects can be obtained by taking 1g of activated charcoal at least 30 minutes before a meal, or 1g immediately after a meal."

[0093] In addition, the website of the Forestry Agency of Japan has an article about new uses for charcoal. For example, by putting charcoal into a bathtub, the bath water becomes alkaline, providing the same bathing effect as a hot spring, and the water itself can also be expected to be purified. Also, by putting charcoal into rivers and lakes, the charcoal's adsorption action removes pollutants, and the microorganisms living in the pores of the charcoal are expected to decompose the pollutants themselves, purifying the water. As is clear from these examples, by using "functional edible charcoal" in various aspects of daily life, such as beverages, bath additives, and cosmetics, it is expected that the wastewater will eventually flow into the wastewater, contributing to the environment in the same way as charcoal.

[0094] In addition, the reduced fermentation lactic acid bacteria according to this embodiment may be powdered for the convenience of storage and transportation, user convenience, etc. That is, after mixing the liquid at the above-mentioned blending ratio, the reduced fermentation lactic acid bacteria can be powdered by adsorbing the liquid to water-soluble dietary fibers such as "inulin", "acacia dietary fiber", "dextrin", and "resistant dextrin" by a method such as spray drying or freeze drying. Here, spray drying is inexpensive, but since high temperatures are applied during production, there is a risk that the raw material properties such as enzymes and polyphenols will be deactivated, and the powder will have a low concentration, so freeze drying is considered to be preferable. Note that the present invention is not limited to these descriptions, and when powdering, commonly known "water-soluble dietary fibers such as pectin and alginic acid", "insoluble dietary fibers such as cellulose and lignin", and "oligosaccharides such as raffinose and fructooligosaccharides" can also be applied.

[0095] The reduced fermented lactic acid bacteria according to this embodiment is constituted by blending the above-mentioned substances, and can obtain a bactericidal effect against so-called "bad bacteria" in the body and an anti-aging effect as described below. In addition to being orally administered, it is also effective to apply it to the outer skin of the human body, and is useful for hair care, face washing, laundry, etc. by mixing it with shampoo, face wash, or detergent.

[0096] [Verification example of bactericidal effect against Streptococcus mutans] A 5-fold dilution of the reduced fermented lactic acid bacteria powder according to the present embodiment was used as a specimen. After the test liquid was inoculated into this specimen, the viable cell count in the test liquid was measured after a predetermined time had elapsed. In addition, a preliminary test (confirmation of neutralization conditions) was conducted in advance to confirm the conditions under which the viable cell count could be measured without being affected by the specimen. The test conditions are as shown in the following table.

[0097] [Table 17]

[0098] Next, the test results are shown. [Table 18] In the table showing the test results, "<10" indicates that no detection was found. The storage temperature was room temperature.

[0099] As described above, the reduced fermented lactic acid bacteria powder according to the present embodiment was found to have a sufficient bactericidal effect against mutans streptococci, since no live bacteria were detected after 5 minutes. This is expected to reduce mutans streptococci, which are bacteria that cause dental caries in the oral cavity, and improve the oral environment, and it is expected to be used for oral care.

[0100] [Verification example of bactericidal effect against Porphyromonas gingivalis] A 5-fold dilution of the reduced fermented lactic acid bacteria powder according to the present embodiment was used as a specimen. After the test liquid was inoculated into this specimen, the viable cell count in the test liquid was measured after a predetermined time had elapsed. In addition, a preliminary test (confirmation of neutralization conditions) was conducted in advance to confirm the conditions under which the viable cell count could be measured without being affected by the specimen. The test conditions are as shown in the following table.

[0101] [Table 19]

[0102] Next, the test results are shown. [Table 20] In the table showing the test results, "<100" indicates that no detection was found. The storage temperature was room temperature.

[0103] As described above, the reduced fermented lactic acid bacteria powder according to the present embodiment was found to have a sufficient bactericidal effect against Porphyromonas gingivalis, since no live bacteria were detected after 1 minute. This is expected to reduce Porphyromonas gingivalis, which is the causative bacterium of periodontal disease in the oral cavity, and improve the oral environment, and is expected to be useful for oral care.

[0104] [Verification example of bactericidal effect against Pasteurella] A 5-fold dilution of the reduced fermented lactic acid bacteria powder according to the present embodiment was used as a specimen. The test liquid was added to this specimen and mixed. The number of remaining viable bacteria was measured immediately after mixing and after reacting at room temperature for a certain period of time. The test conditions are as shown in the following table.

[0105] [Table 21]

[0106] Next, the test results are shown. [Table 22] In the table showing the test results, "<100" indicates that no detection was found. The storage temperature was room temperature.

[0107] As described above, the reduced fermented lactic acid bacteria powder according to the present embodiment was found to have a sufficient bactericidal effect against Pasteurella bacteria, since no live bacteria were detected after 1 minute. This is expected to reduce the number of Pasteurella bacteria, which are oral bacteria that infect humans from dogs and cats, and improve the oral environment, and it is expected to be used for oral care.

[0108] [Verification example of bactericidal effect against Helicobacter pylori] A 5-fold dilution of the reduced fermented lactic acid bacteria powder according to this embodiment was used as a specimen. A test bacteria liquid (hereinafter referred to as "test liquid") was inoculated into this specimen, and the number of live bacteria in the test liquid was measured after a predetermined time had passed. In addition, a preliminary test (confirmation of neutralization conditions) was conducted in advance to confirm the conditions under which the number of live bacteria could be measured without being affected by the specimen. The test conditions are as shown in the following table.

[0109] [Table 23]

[0110] Next, the test results are shown. [Table 24] In the table showing the test results, "<1000" indicates that no detection was found. The storage temperature was room temperature.

[0111] Thus, in the reduced fermented lactic acid bacteria powder according to the present embodiment, the number of live bacteria was 1 / 350 of the control after 1 minute, and was not detected after 5 minutes, demonstrating a sufficient bactericidal effect against H. pylori. As a result, drinking the powder is expected to reduce the number of H. pylori, which is a cause of stomach cancer and stomach ulcers, and improve the stomach environment.

[0112] [Verification example of bactericidal effect against Staphylococcus aureus] A 5-fold dilution of the reduced fermented lactic acid bacteria powder according to the present embodiment was used as a specimen. After the test liquid was inoculated into this specimen, the viable cell count in the test liquid was measured after a predetermined time had elapsed. In addition, a preliminary test (confirmation of neutralization conditions) was conducted in advance to confirm the conditions under which the viable cell count could be measured without being affected by the specimen. The test conditions are as shown in the following table.

[0113] [Table 25]

[0114] Next, the test results are shown. [Table 26] In the table showing the test results, "<10" indicates that no detection was found. The storage temperature was room temperature.

[0115] Thus, it was revealed that the reduced fermented lactic acid bacteria powder according to this embodiment has a sufficient bactericidal effect against Staphylococcus aureus, since no live bacteria were detected after 5 minutes. This is expected to reduce the bad intestinal bacteria that increase with age, i.e., Staphylococcus aureus, one of the two major bad bacteria in the large intestine, and lead to an intestinal flora dominated by good bacteria. Staphylococcus aureus is said to be a causative bacterium of atopic dermatitis on the skin and scalp, and by mixing the reduced fermented lactic acid bacteria powder according to this embodiment with hair shampoo, hair treatment, facial cleanser, etc., it is expected to be used for skin care and scalp care. That is, it can be used for face washing, oral care, scalp care, or added to skin gel packs, bath additives, etc. By mixing the reduced fermented lactic acid bacteria powder according to this embodiment with shampoo or facial cleanser, it is possible to easily replenish hydrogen and minerals to the scalp and skin, and reduce the skin cleanly to suppress oxidation troubles. As described above, since the reduced fermented lactic acid bacteria powder according to this embodiment has a high bactericidal effect, it is expected to be effective against Propionibacterium acnes and Malassezia fungus, and it is expected to be useful for treating acne and folliculitis.

[0116] [Verification example of bactericidal effect against Clostridium perfringens] A 5-fold dilution of the reduced fermented lactic acid bacteria powder according to the present embodiment was used as a specimen. The test liquid was added to this specimen and mixed. After a predetermined time had elapsed, the viable cell count in the test liquid was measured. The test conditions were as shown in the following table.

[0117] [Table 27]

[0118] Next, the test results are shown. [Table 28] In the table showing the test results, "<10" indicates that no detection was found. The storage temperature was room temperature.

[0119] Thus, it was revealed that the reduced fermented lactic acid bacteria powder according to this embodiment has a sufficient bactericidal effect against Clostridium perfringens, since no live bacteria were detected after 1 minute. In the human intestine, changes characterized by a decrease in bifidobacteria and an increase in Clostridium perfringens occur from middle age onwards. Clostridium perfringens is one of the putrefactive bacteria, which putrefies proteins to produce harmful substances such as ammonia, amines, phenols, and indoles. These harmful substances also include carcinogens, most of which are broken down in the liver, but if they exceed the amount the liver can process, they will affect the entire body. Indole is converted to indoxyl sulfate in the liver. Although this is not a problem for people with healthy kidneys, as they can excrete indoxyl sulfate from urine, chronic kidney disease patients, who are said to number one in eight Japanese people and approximately 13 million in Japan, are unable to excrete indoxyl sulfate from urine, and research is currently being conducted on the fact that indoxyl sulfate flows into the bloodstream and is the causative substance that causes uremia and heart disease. This decrease in bifidobacteria and relative increase in Clostridium perfringens can be considered as aging in the intestine. The reduced fermented lactic acid bacteria powder according to this embodiment is expected to have the effect of reducing bad intestinal bacteria that increase with age, i.e., Clostridium perfringens, one of the two major bad bacteria in the large intestine, and leading to an intestinal flora dominated by good bacteria.

[0120] [A case study on the "bactericidal effect on oral bacteria" of a man in his 70s with periodontal disease] The specimen was the reduced fermented lactic acid bacteria powder according to the present embodiment. The test liquid was the "saliva of a man in his 70s with periodontal disease." As controls, gels containing only KS Melt lactic acid bacteria production substances, with and without mint, were used. That is, "KS Melt Gel with Mint" and "KS Melt Gel without Mint." In addition, "Control Sample 1," a commercially available toothpaste, was used. The test conditions are as shown in the following table. That is, (1) the test liquid was diluted 100 times with physiological saline, and then filtered (Advantec filter paper No. 2), and (2) 1 g of the specimen was added to 10 mL of the liquid obtained in (1) above, mixed for 3 minutes, and then inoculated into the medium after dilution. The medium used was Nissui Compact Dry TC manufactured by Nissui Pharmaceutical Co., Ltd. The results are as follows.

[0121] [Table 29] In the test result tables, "<10" means that no detection was detected, and ">99" means that the result is as close to 100% as possible.

[0122] Thus, although a reduction in the number of bacteria was observed in "KS Melt Gel with Mint", "KS Melt Gel without Mint" and Control Sample 1, a considerable proportion of bacteria remained, whereas no bacteria were detected in the reduced fermented lactic acid bacteria powder according to this embodiment, demonstrating that bacteria could be almost completely eradicated. This demonstrates that the reduced fermented lactic acid bacteria powder according to this embodiment has a sufficient bactericidal effect on oral bacteria.

[0123] [Verification example of existing live bacteria] The samples used were "KS Melt Gel with mint", "KS Melt Gel without mint" and the reduced fermented lactic acid bacteria powder according to this embodiment. 1 g of each sample was added with physiological saline to make 5 g, and the medium was inoculated with Nissui Compact Dry TC manufactured by Nissui Pharmaceutical Co., Ltd. The results are as follows.

[0124] [Table 30]

[0125] In this way, it was possible to confirm that no live bacteria were present in "KS Melt Gel with Mint", "KS Melt Gel without Mint", and the reduced fermented lactic acid bacteria powder according to this embodiment.

[0126] [Example of verification of antioxidant properties (SOD-like activity)] The reduced fermented lactic acid bacteria powder according to this embodiment was diluted 100 times with pure water and filtered through a 0.45 μm filter to prepare a stock solution. The stock solution and the stock solution were diluted 10 times and 100 times with the buffer solution included with the SOD Assay Kit-WST (manufactured by Dojindo Chemical Industries, Ltd.) to calculate the superoxide production inhibition rate as SOD-like activity (%) according to a predetermined method using the SOD Assay Kit-WST. The results are as follows, and the higher the SOD-like activity, the higher the antioxidant property.

[0127] [Table 31]

[0128] Thus, the reduced fermented lactic acid bacteria powder according to this embodiment has high SOD-like activity in the original solution. In addition, even when diluted 10 or 100 times, it has a sufficiently high SOD-like activity, so it is expected to be sufficiently effective even when diluted, and it was found that it has a sufficiently high antioxidant property (SOD-like activity) from the viewpoint of practical use. This can be expected to have an antioxidative effect on the body caused by active oxygen.

[0129] [Example of verification of α-glucosidase inhibitory activity] We verified the α-glucosidase inhibitory activity (α-glucosidase inhibitory activity measurement). α-glucosidase creates a state of high blood sugar levels in the body, so if the α-glucosidase inhibitory activity is high, it will inhibit the absorption of sugar in the body and avoid a state of high blood sugar levels, which will ultimately suppress the production of AGEs.

[0130] The reduced fermented lactic acid bacteria powder according to this embodiment was diluted 100 times with pure water and filtered through a 0.45 μm filter to prepare the stock solution. The stock solution and the liquids obtained by diluting the stock solution 10 times and 100 times with pure water were used as the measurement samples. After adding 10 μL of sample solution to each microtube and stirring, 40 μL of enzyme solution and 40 μL of 50 mM sodium phosphate buffer were added, and the mixture was pre-incubated at 37 ° C for 5 minutes, and then 950 μL of substrate solution was added and stirred, followed by incubation at 37 ° C for 15 minutes. Then, 1000 μL of 0.5 M Tris aqueous solution was added to stop the reaction, and the absorbance at 405 nm was measured. The α-glucosidase inhibitory activity (%) when the sample was added was calculated from the change in absorbance. The results are as follows. [Table 32]

[0131] Thus, it was found that the reduced fermented lactic acid bacteria powder according to this embodiment has extremely high α-glucosidase inhibitory activity. This suggests that by inhibiting α-glucosidase activity in vivo, it is possible to inhibit sugar absorption and avoid high blood sugar levels. If high blood sugar levels can be avoided, it will be possible to create a state in which early AGEs-producing reaction products are less likely to be produced.

[0132] [Verification example of inhibitory activity against the formation of amadori compounds] In the glycation reaction in the body, Amadori compounds are produced as "early AGE-forming reactants," so if the production of Amadori compounds is inhibited, the production of the next intermediate reactants and the subsequent late AGE-forming reactants is inhibited, resulting in the inhibition of the production of AGEs. As blood sugar levels rise, "hemoglobin, a protein present in red blood cells," undergoes glycation and turns into "glycated hemoglobin." "Glycated hemoglobin" is a type of Amadori compound, and the test item used to examine this is "hemoglobin A1c," which is used as a criterion for diagnosing diabetes.

[0133] The reduced fermented lactic acid bacteria powder according to the present embodiment was diluted 100 times with pure water and filtered through a 0.45 μm filter to prepare a stock solution. The stock solution and the stock solution diluted 10 times and 100 times with pure water were used as measurement samples. The measurement of the amadori compound production inhibitory activity was performed by mixing the measurement sample, glucose solution, and bovine serum albumin solution and incubating at 60°C for 48 hours to prepare a saccharification reaction solution. 300 μL of 0.25 mM nitrobrute tetrazolium solution (pH 9.0) was added to 40 μL of the saccharification reaction solution, and the mixture was incubated at 37°C for 30 minutes, and then allowed to stand at room temperature for 16 hours, after which the absorbance at 540 nm was measured. The amadori compound production inhibitory activity when the measurement sample was added was calculated from the change in absorbance. The results are as follows.

[0134] [Table 33] In the above table, negative numbers are equivalent to 0.

[0135] Thus, the reduced fermented lactic acid bacteria powder according to the present embodiment has a high inhibition rate in the original solution. Therefore, it can be said that the reduced fermented lactic acid bacteria powder according to the present embodiment has an activity of inhibiting the production of Amadori compounds, and therefore, when consumed, it is expected to have a glycation inhibitory effect that lowers the value of "hemoglobin A1c".

[0136] [Example of verification of AGEs cross-linking cleavage function] The AGEs cross-linking cleavage function was verified by measuring the amount of benzoic acid produced by decomposition of the dicarbonyl bond of the model compound 1-phenyl-1,2-propanedione, which has a dicarbonyl bond, when the dicarbonyl bond is cleaved. The reduced fermented lactic acid bacteria powder according to this embodiment was diluted 100 times with pure water and filtered through a 0.45 μm filter to prepare a stock solution. This stock solution and solutions obtained by diluting the stock solution 10 times and 100 times with pure water were used as measurement samples.

[0137] The measurement sample, 1-phenyl-1,2-propanedione solution, and methanol were mixed to prepare a reaction solution. The reaction solution was incubated at 37°C for 24 hours, and then hydrochloric acid was added to stop the reaction. The amount of benzoic acid produced by decomposition in the reaction solution was quantified using high performance liquid chromatography. The AGEs cross-link cleavage rate was calculated as 100% when all 1-phenyl-1,2-propanedione in the reaction solution was decomposed to benzoic acid. The results are as follows:

[0138] [Table 34]

[0139] As a result, the reduced fermentation lactic acid bacteria powder according to this embodiment has an AGE cross-linking cleavage function, and is expected to have the effect of decomposing the intermediate reaction product "AGE cross-linking" that has already been generated, thereby suppressing glycation.

[0140] [Verification example of fluorescent AGEs production inhibitory activity] The reduced fermented lactic acid bacteria powder according to this embodiment was diluted 100-fold with pure water and filtered through a 0.45 μm filter to prepare a stock solution. This stock solution and the stock solution diluted 10-fold and 100-fold with pure water were used as measurement samples. The fluorescent AGEs production inhibitory activity was measured by mixing the measurement sample, glucose solution, and bovine serum albumin solution and incubating at 60°C for 40 minutes, after which the generated fluorescent AGEs were measured with a fluorescence spectrophotometer at an excitation wavelength of 370 nm and a fluorescence wavelength of 440 nm. The fluorescent AGEs production inhibitory activity (%) when the measurement sample was added was calculated as the inhibition rate from the decrease in fluorescence intensity. The results are as follows:

[0141] [Table 35]

[0142] Thus, the reduced fermented lactic acid bacteria powder according to this embodiment has a high inhibition rate in its original form. Even when diluted 10 or 100 times, the inhibition rate is still sufficiently high, so it is expected to be effective even when diluted, and it has been found to have a sufficiently high inhibitory effect on the production of late-stage AGEs, "fluorescent AGEs," from the practical standpoint. As a result, it can be expected to have an inhibitory effect on glycation when consumed.

[0143] [Verification example of tyrosinase inhibitory activity] The reduced fermented lactic acid bacteria powder according to this embodiment was diluted 100 times with pure water and filtered through a 0.45 μm filter to prepare a stock solution. This stock solution and the stock solution diluted 10 times and 100 times with pure water were used as measurement samples. For the measurement, a solution of tyrosinase derived from mushrooms dissolved at 300 units / mL in 0.1 M phosphate buffer (pH 6.7) was used as an enzyme solution, and a solution of L-DOPA dissolved at 1 mM in 0.1 M phosphate buffer (pH 6.7) was used as a substrate solution.

[0144] 25 μL of sample solution and 100 μL of enzyme solution were placed in a 96-well microplate and incubated at 37°C for 10 minutes, after which 125 μL of substrate solution was added and incubated at 37°C for 30 minutes, and the absorbance at 490 nm was measured. The tyrosinase inhibitory activity (%) upon addition of the sample was calculated from the change in absorbance. The results are as follows:

[0145] [Table 36]

[0146] Tyrosinase is known to produce melanin in the body, so if it is present in large quantities, it will increase skin blemishes. The reduced fermented lactic acid bacteria powder according to this embodiment has high tyrosinase inhibitory activity in its original form, but when diluted 10 times, it showed a higher value. This shows that sufficient effect can be expected even when diluted, and that a sufficiently high tyrosinase inhibitory activity function can be obtained from the viewpoint of practicality. This makes it difficult for melanin to be synthesized, and is expected to prevent skin blemishes.

[0147] [Verification example of hyaluronidase inhibitory activity] The reduced fermented lactic acid bacteria powder according to this embodiment was diluted 100 times with pure water and filtered through a 0.45 μm filter to prepare a stock solution. This stock solution and the liquids obtained by diluting the stock solution 10 times and 100 times with pure water were used as measurement samples. For the measurement, hyaluronidase was dissolved at 400 units / mL in 0.1 M acetate buffer (pH 4.0) to prepare an enzyme solution, potassium hyaluronate was dissolved at 1 mg / mL in acetate buffer (pH 4.0) to prepare a substrate solution, Compound48 / 80 was dissolved at 0.1 mg / mL in acetate buffer (pH 4.0) to prepare an enzyme activator, and dimethylaminobenzaldehyde was dissolved at 10 mg / mL in a hydrochloric acid and acetic acid mixture to prepare a color developing solution.

[0148] 12 μL of sample solution and 12 μL of enzyme solution were added to a microtube and incubated at 40° C. for 20 minutes, then 12 μL of enzyme activator was added and incubated at 40° C. for 20 minutes. 12 μL of substrate solution was added and incubated at 40° C. for 40 minutes, then 12 μL of 0.4NaOH aqueous solution was added and cooled on ice. Then, 12 μL of 0.8M borate buffer (pH 9.0) was added, boiled for 3 minutes, cooled on ice, 180 μL of color developing solution was added, incubated at 40° C. for 30 minutes, and absorbance at 595 nm was measured. Hyaluronidase inhibitory activity (%) when the sample was added was calculated from the change in absorbance. The results are as follows.

[0149] [Table 37]

[0150] Hyaluronic acid is said to exist in large amounts between cells, and protects cells by its water retention and buffering functions. On the other hand, hyaluronidase is an "enzyme that breaks down hyaluronic acid" that increases due to aging and UV stimulation, and if it is present in large amounts, it is impossible to maintain the moisture retention and viscoelasticity of the skin, which may lead to wrinkles and sagging of the skin. The reduced fermented lactic acid bacteria powder according to this embodiment has high hyaluronidase inhibitory activity in its original form. In addition, even if it is diluted 10 times or 100 times, it has a sufficiently high hyaluronidase inhibitory activity, so it is expected that it will have a sufficient effect even if it is diluted, and it has been found that a sufficiently high hyaluronidase inhibitory activity function can be obtained from the viewpoint of practicality. This makes it difficult for hyaluronic acid to be decomposed, and it is expected that the moisture retention and viscoelasticity of the skin will be maintained, preventing wrinkles and sagging of the skin.

[0151] [Verification example of collagenase inhibitory activity] Collagenase is a proteolytic enzyme that hydrolyzes collagen. The present inventors conducted a test on the "collagenase inhibitory activity" of the reduced fermented lactic acid bacteria powder according to this embodiment. The test method was to dissolve the reduced fermented lactic acid bacteria powder in pure water to prepare a 10 mg / mL solution, which was then filtered through a 0.45 μm filter to prepare a stock solution. The stock solution and solutions obtained by diluting the stock solution 10-fold and 100-fold with pure water were used as measurement samples.

[0152] For the measurement, Pz-peptide was dissolved in 0.1 M Tris buffer (pH 7.1) at 0.39 mg / mL to prepare a substrate solution, and collagenase Type IV was dissolved in pure water at 0.1 mg / mL to prepare an enzyme solution. 25 μL of sample solution, 25 μL of enzyme solution, and 200 μL of substrate solution were placed in a glass test tube, and incubated at 37°C for 30 minutes. Then, 250 μL of 25 mM citric acid was added to stop the reaction. Then, 2500 μL of ethyl acetate was added and the mixture was shaken for extraction, followed by centrifugation (1500 rpm, 5 minutes), and the upper layer (ethyl acetate layer) was collected and the absorbance at 320 nm was measured. The collagenase inhibitory activity when the sample was added was calculated from the change in absorbance, and was taken as the collagenase inhibitory activity. The test results are shown in the table below. [Table 38] In the above table, negative numbers are equivalent to 0.

[0153] In this way, it has been revealed that the reduced fermented lactic acid bacteria powder according to this embodiment has the function of inhibiting collagenase, which breaks down collagen. Therefore, it is expected that collagen in the skin will be less likely to be broken down by ingesting or applying the reduced fermented lactic acid bacteria according to this embodiment.

[0154] [Verification example of elastase inhibitory activity] Elastase is an enzyme that breaks down elastin, an elastic fiber that determines the mechanical properties of connective tissue together with collagen. The present inventors conducted a test on the "elastase inhibitory activity" of the reduced fermented lactic acid bacteria powder according to this embodiment. The test method was to dissolve the reduced fermented lactic acid bacteria powder in pure water to prepare a 10 mg / mL solution, which was then filtered through a 0.45 μm filter to prepare a stock solution. The stock solution and solutions obtained by diluting the stock solution 10-fold and 100-fold with pure water were used as measurement samples.

[0155] For the measurement, STANA (N-Succinyl-Ala-Ala-Ala-p-nitroanilide) was dissolved at 1 mM in 50 mM Tris buffer (pH 8.8) to prepare a substrate solution, and porcine pancreatic elastase was dissolved at 0.5 units / mL in 50 mM Tris buffer (pH 8.8) to prepare an enzyme solution. Using a 96-well microplate, 50 μL of sample solution, 50 μL of enzyme solution, and 100 μL of substrate solution were placed and incubated at 37°C for 30 minutes, after which the absorbance at 405 nm was measured. The elastase inhibitory activity when the sample was added was calculated from the change in absorbance, and was taken as the elastase inhibitory activity. The test results are shown in the table below. [Table 39] In the above table, negative numbers are equivalent to 0.

[0156] Thus, it has been revealed that the reduced fermented lactic acid bacteria powder according to this embodiment has the function of inhibiting elastase, which breaks down elastin. Therefore, it is expected that by ingesting or applying the reduced fermented lactic acid bacteria according to this embodiment, elastin in the skin will be less likely to be broken down.

[0157] [Verification example of lipase inhibitory activity] The reduced fermented lactic acid bacteria powder according to this embodiment was diluted 100 times with pure water and filtered through a 0.45 μm filter to prepare a stock solution. This stock solution and the liquids obtained by diluting the stock solution 10 times and 100 times with pure water were used as measurement samples. The measurement was performed using a Lipase Kit S (DS Pharma Biomedical) with some improvements to the operating procedure. The measurement sample, lipase solution, substrate solution, and color development solution were mixed and incubated at 30° C. for 30 minutes, after which a stop solution was added and the absorbance at 405 nm was measured. The lipase inhibitory activity (%) when the sample was added was calculated from the change in absorbance. The results are as follows.

[0158] [Table 40]

[0159] Lipase is an enzyme that breaks down and emulsifies lipids, one of the three major nutrients, and is mainly secreted in the stomach. The emulsified oil moves to the small intestine, where it is broken down into small pieces by the action of bile acids and absorbed by the body. If the decomposition (emulsification) of oil in food by lipase can be suppressed, it will not be absorbed by the body and will move to the large intestine and be excreted with feces, so a diet effect is expected. The reduced fermented lactic acid bacteria powder according to this embodiment shows a lipase inhibitory activity of 18.1% in the original solution, and is expected to have a diet effect in that it inhibits lipase, an enzyme that breaks down lipids, and makes it difficult for the body to absorb fat.

[0160] [Example of verification of effects on lactic acid bacteria] As described above, the reduced fermented lactic acid bacteria powder according to this embodiment has a high bactericidal ability against bad bacteria, and is therefore effective in removing so-called "bad bacteria." However, at the same time, we investigated whether it also has an effect on the so-called "good bacteria," lactic acid bacteria (Lactobacillus plantarum).

[0161] The reduced fermented lactic acid bacteria powder according to this embodiment was used as a "specimen," and the specimen was diluted to 1% with sterilized water to prepare a test sample. The test bacteria (lactic acid bacteria: Lactobacillus plantarum) was inoculated onto a BCP agar medium and cultured at 30°C for 48 hours. After the culture, the number of bacteria was reduced to "10" using sterilized physiological saline. 8 The test bacteria liquid was prepared by adjusting the concentration of the test sample to "1 / mL". Then, 0.1 mL of the test bacteria liquid was inoculated into 10 mL of the test sample and cultured at 25°C. Five minutes and 15 minutes after inoculation, a 10-fold serial dilution of the test sample was made using sterilized saline to prepare test liquids, which were then inoculated onto BCP agar medium and cultured at 30°C for 48 to 72 hours. After culture, the formed colonies were counted and the viable cell count was calculated. The test was also carried out in the same manner using sterilized phosphate buffered saline as a control. The results are as follows. [Table 41] In the table showing the test results above, the figures indicate the average viable cell count (CFU / mL) and n3.

[0162] Thus, it was found that the reduced fermented lactic acid bacteria powder according to this embodiment did not reduce the number of live lactic acid bacteria even after 15 minutes. This revealed that the reduced fermented lactic acid bacteria powder according to this embodiment has an effective bactericidal function against bad bacteria, but does not affect good bacteria. The reduced fermented lactic acid bacteria powder according to this embodiment is made from good bacteria (live bacteria) and their secretions, plum extract, deep sea water, and hydrogen, and the above test results suggest that it is unlikely to have a negative effect on good bacteria (live bacteria), i.e., bacteria that are beneficial to the body, such as bifidobacteria and lactic acid bacteria.

[0163] On the other hand, periodontal disease is a disease that is less painful than tooth decay, progresses slowly, and is said to affect 80% of people by age 30 and 90% by age 50, so anyone can get it. Porphyromonas gingivalis, the main cause of periodontal disease, spreads in the oral cavity, creating an environment where bad bacteria dominate. Therefore, to improve the oral environment, chewables and gums containing good bacteria (live bacteria) have been released one after another. It is important to provide good bacteria (live bacteria) in the oral cavity, and although the image is good, the rate at which good bacteria (live bacteria) settle in the oral cavity is not high. For this reason, it is also important to directly act on Porphyromonas gingivalis and Streptococcus mutans to reduce the dominant bad bacteria. In that sense, the collaboration between good bacteria (live bacteria) and reduced fermentation lactic acid bacteria can be expected to have a synergistic effect on the approach to oral care.

[0164] KT-11 lactic acid bacteria (dead bacteria) is well-known as a lactic acid bacteria with a strong evidence base for oral care, and oral chewables and supplements containing KT-11 lactic acid bacteria are on the market. KT-11 lactic acid bacteria (dead bacteria) are crispatus bacteria, which are abundant in the birth canal of women and are the first lactic acid bacteria that babies receive at birth. They have been confirmed to have the function of increasing components that protect the intestinal mucosa, but they do not directly act on bacteria in the oral cavity. When swallowed, they increase intestinal immune activity, and as an indirect effect, they increase the IgA antibodies in saliva, which enhances the bactericidal power of saliva and leads to a mechanism for reducing Porphyromonas gingivalis, a bad bacterium in the oral cavity. For this reason, oral chewables and supplements containing good bacteria, whether live or dead, are thought to enhance intestinal immunity by swallowing them, and as a result, they increase the IgA antibodies in saliva, which indirectly has a positive effect on the oral environment. The reduced fermented lactic acid bacteria powder according to this embodiment is a lactic acid bacteria fermentation extract based on "KS melt lactic acid bacteria production material" which has been used for many years in dentist research groups and has a wealth of evidence for oral care. At the same time, it contains "30 billion to 150 billion / g" of good bacteria (dead bacteria). In addition to a direct effect on bad bacteria in the oral cavity, it is expected to increase "IgA antibodies" in saliva through improved intestinal immunity. Furthermore, the above-mentioned "KT-11 lactic acid bacteria" has been patented to increase the synergistic effect with other good bacteria (Patent No. 4942831), and by simultaneously blending the reduced fermented lactic acid bacteria and "KT-11 lactic acid bacteria", it is expected to further increase the synergistic effect. Figure 15 is a diagram showing the test results of the synergistic effect between "KS melt lactic acid bacteria production material (raw material that is the base of reduced fermented lactic acid bacteria)" and "KT-11 lactic acid bacteria HP". This test uses the samples "KS Melt Lactic Acid Bacteria Production Material: manufactured by KS West Japan Co., Ltd." and "KT-11 Lactic Acid Bacteria HP: manufactured by Kitty Co., Ltd.", which are the base raw materials for reduced fermentation lactic acid bacteria, to evaluate the interleukin 12 (IL-12) production induction ability using animal immune cells. This "KT-11 Lactic Acid Bacteria HP: manufactured by Kitty Co., Ltd." is composed of a mixed raw material of "20% KT-11 Lactic Acid Bacteria (dead bacteria) and 80% dextrin," and is known to have an immune activation (biogenics) function by stimulating the intestinal tract. The test method is as follows. That is, the cells used were macrophage-like cell line (immune cells), and the cultured cells were suspended in a medium, then dispensed into a microplate, and the test substance was added to the final concentration, and the amount of IL-12 produced in the culture supernatant was measured by ELISA. The test results are shown in Figure 15. As shown in Figure 15, the "immunoactive IL-12 value" of "KT-11 lactic acid bacteria HP" alone was 1847 (pg / mL), but when the same amount of "KT-11 lactic acid bacteria HP" was added to "KS melt lactic acid bacteria production material (raw material that serves as the base for reduced fermentation lactic acid bacteria)", the "immunoactive IL-12 value" was 2515 (pg / mL), indicating a synergistic effect of more than 36%.

[0165] The present inventors have confirmed the function of the reduced fermented lactic acid bacteria powder according to the present embodiment by referring to the results of the verification of the effects of the deep sea water according to the present embodiment on bacteria such as lactic acid bacteria.

[0166] [Verification example of bacterial growth in commercially available yogurt] The present inventors focused on the effect of the deep sea water according to this embodiment on the proliferation of bacteria in commercially available yogurt, and confirmed the results of a test conducted by a third party. The test method was as follows: (a) 1 mL of commercially available yogurt A and B diluted 100 times was added to 12 mL of MRS medium or MRS medium to which 1% deep sea water powder had been added, (b) the culture was conducted at 37°C, and the turbidity was measured over time. This "turbidity" indicates the degree of turbidity of a liquid, and the degree of turbidity when 1 mg of white clay is contained in 1 L of distilled water is defined as 1 degree (or 1 ppm). Figure 1 shows the effect of deep sea water powder on the proliferation of bacteria in commercially available yogurt A, and Figure 2 shows the effect of deep sea water powder on the proliferation of bacteria in commercially available yogurt B. As shown in Figures 1 and 2, it has been revealed that the addition of deep sea water powder promotes the proliferation of bacteria in yogurt.

[0167] [Verification example of the proliferation of "Lactobacillus gasseri JCM1131" bacteria] Conventionally, the results of a verification of the effect of deep sea water on the proliferation of bacteria "Lactobacillus gasseri JCM1131) known as intestinal lactic acid bacteria are known. The present inventors confirmed the function of the reduced fermented lactic acid bacteria powder according to this embodiment by referring to the verification results. In this test method, (a) 1 mL of "Lactobacillus gasseri JCM1131" strain with a turbidity (OD600) value of 1.0 (MRS medium) was added to 12 mL of modified MRS medium or modified MRS medium to which deep sea water powder was added at 1% or 2%, and (b) the mixture was cultured at 37°C, and the turbidity was measured over time. Figure 3 shows the effect of adding deep sea water powder on the proliferation of "Lactobacillus gasseri JCM1131". As shown in Figure 3, it has been revealed that the addition of deep sea water powder promotes the proliferation of the "Lactobacillus gasseri JCM1131" strain.

[0168] In this way, by combining deep sea water with lactic acid bacteria fermentation extract, it is expected that drinking the reduced fermented lactic acid bacteria powder will be beneficial for intestinal lactic acid bacteria.

[0169] [Verification example of total peptide quantification for reduced fermented lactic acid bacteria powder] The present inventors conducted the following test for the total peptide quantification of reduced fermented lactic acid bacteria powder. The test method was to dissolve reduced fermented lactic acid bacteria powder in pure water, adjust the concentration to 5 mg / mL, and filter the resultant through a 0.45 μm filter to prepare a measurement sample. The measurement sample was subjected to total peptide quantification using "Pierce Quantitative Colorimetric Peptide Assay (manufactured by Thermo Fisher Scientific)". The results are shown in the following table.

[0170] [Table 42] This result shows the total peptide amount per 100 g of the reduced fermented lactic acid bacteria powder. Thus, it was found that the reduced fermented lactic acid bacteria powder according to this embodiment contains a very large amount of peptides.

[0171] [Verification example of deodorizing properties] The present inventors conducted a test on the deodorizing properties of the reduced fermented lactic acid bacteria powder according to this embodiment. In this deodorizing property test, the deodorizing properties of the reduced fermented lactic acid bacteria powder according to this embodiment were verified for "ammonia" which causes bad odors such as sweat odor, aging odor, excrement odor, cigarette odor, food waste odor, and ammonia odor, and "trimethylamine" which emits the odor of rotten fish. The specimen was the reduced fermented lactic acid bacteria powder according to this embodiment, and a 5-fold diluted solution of the reduced fermented lactic acid bacteria powder was used, which was produced by increasing the amount of hydrogen and deep sea water added during reduction fermentation by hydrogen by more than twice the amount of the reduced fermented lactic acid bacteria powder used in the above-mentioned verification examples of mutans and Porphyromonas gingivalis. The test method was to dilute the sample 5 times with distilled water, use ammonia (100 ppm) and "trimethylamine (28 ppm)" as odor components, set the sample volume to 0.5 mL, and set the standing time to 30 minutes and 120 minutes. The test results are as shown in the following table.

[0172] [Table 43]

[0173] It was found that the deodorizing effect was observed even with water alone compared to the blank, but in contrast, the reduced fermented lactic acid bacteria powder SP1 and reduced fermented lactic acid bacteria powder SP2 according to the present embodiment exhibited a high reduction rate (deodorizing rate) of 98% or more for both "ammonia" and "trimethylamine". In addition, it was found that the reduced fermented lactic acid bacteria powder SP1 and reduced fermented lactic acid bacteria powder SP2 according to the present embodiment have a higher deodorizing function for "ammonia" than "Ina Akamatsu Myo-tan / coconut shell activated carbon granules (dispersed processing)", which is generally said to have a high deodorizing effect, and also have a deodorizing function that is comparable to "persimmon tannin extract", which is famous for its high deodorizing effect. From the above, it is expected to reduce the bad odor itself generated in the oral cavity and intestines, and to improve the oral environment and gastrointestinal environment, and it can be expected to be used for oral care and gastrointestinal care. It is known that "Helicobacter pylori" exists in the stomach while neutralizing gastric acid by producing an enzyme called urease and creating alkaline ammonia around itself. As mentioned above, reduced fermentation lactic acid bacteria eliminate Helicobacter pylori while in collaboration with functional edible charcoal such as Ina Akamatsu Myotan and coconut shell activated charcoal granules (dispersed processing), which have high adsorption activity for the ammonia produced by Helicobacter pylori through the urease enzyme, we can expect a total approach to Helicobacter pylori, which is believed to be the cause of stomach cancer.

[0174] [Verification example of bactericidal effect against Candida] "Candida bacteria" is neither good nor bad bacteria, but an "opportunistic bacteria" and a type of mold. When the human body's immune system weakens, it grows rapidly, causing oral candidiasis, and proliferating in the intestines, causing intestinal leakage (leaky gut syndrome). The inventors conducted a test on the bactericidal effect of the reduced fermented lactic acid bacteria powder according to this embodiment on Candida bacteria. The specimen was the reduced fermented lactic acid bacteria powder according to this embodiment, and a 5-fold dilution of "reduced fermented lactic acid bacteria powder produced by increasing the amount of hydrogen and deep sea water added at the time of reduction fermentation by hydrogen by more than twice the amount of reduced fermented lactic acid bacteria powder used in the above-mentioned verification examples of mutans and Porphyromonas gingivalis" was used. This is referred to here as "reduced fermented lactic acid bacteria powder SP2". The test bacteria was Candida bacteria (Candida albicans: NBRC1594). The test method is as follows. That is, the specimen was diluted with sterilized water to 1%, 10%, and 20% to prepare the test samples. The test bacteria was inoculated onto potato dextrose agar medium and cultured at 25°C for 48 hours. After the culture, the number of bacteria was reduced to 10 using sterilized physiological saline. 8 The test solution was prepared by adjusting the concentration of the test sample to 100 / mL. Next, 0.1 mL of the test solution was inoculated into 10 mL of the test sample and incubated at 25°C. The 1% solution was prepared by adjusting a 10-fold dilution series of the test sample with sterilized saline 30 minutes and 1 hour after inoculation, and the 10% and 20% solutions were prepared 5 minutes, 15 minutes, 30 minutes and 1 hour after inoculation. These test solutions were inoculated into Sabouraud dextrose agar medium with a neutralizing agent and incubated at 25°C for 48 to 72 hours. After incubation, the formed colonies were counted and the viable cell count was calculated. The test was also conducted in the same manner using sterilized phosphate buffered saline as a control. The test results are shown in the table below.

[0175] [Table 44] The values ​​in the above table indicate the average value of viable bacteria count (CFU / mL) and n3, and "NT" indicates that the test was not performed. As shown in this table, it was found that the reduced fermented lactic acid bacteria powder according to this embodiment does not completely kill Candida bacteria, but can reduce the number of bacteria to a maximum of 1 / 143 of the initial number of bacteria. A high bacterial reduction rate is shown at all concentrations. It was found that the powder has a high bactericidal effect on Candida bacteria, which is a type of mold and is not easy to inhibit from growing. From the above, it can be expected that the powder will have a beneficial effect on the symptoms of oral candidiasis and leaky gut syndrome through its effect of reducing Candida bacteria, which are the cause of these conditions.

[0176] [Ionized apatite blend] The reduced fermented lactic acid bacteria powder according to this embodiment may contain an oral composition containing "hydroxyapatite" and citric acid (hereinafter referred to as "ionized apatite").

[0177] As for hydroxyapatite, it is preferable to use hydroxyapatite derived from scallop shells (bioapatite: registered trademark, defined as naturally derived from living organisms and with low crystallinity). Hydroxyapatite is a component that accounts for approximately 97% of human tooth enamel and approximately 65% ​​of bone, and is composed of phosphoric acid and calcium. It is said to have high biocompatibility, is neutral to weakly alkaline, and is safe for the human body. Hydroxyapatite is almost the same component as tooth enamel, so it has the function of remineralizing and repairing scratches on the surface of teeth caused by eating or brushing teeth, and early caries caused by demineralization. Note that mineral-derived hydroxyapatite may be used instead of natural living organism-derived hydroxyapatite.

[0178] Hydroxyapatite also has 10 calcium atoms per molecule, and these calcium atoms are replaced by various metals such as cadmium and lead through ion exchange. This ion exchange property allows it to adsorb harmful substances such as heavy metals. Hydroxyapatite also has a complex crystal structure and an electrically charged surface, allowing it to adsorb bacteria, viruses, pollen, and the like. Hydroxyapatite also has the function of adsorbing pigments and lipid peroxides.

[0179] Hydroxyapatite that is generally available is often synthesized using a mineral (apatite) as a raw material, but the ionized apatite according to the present embodiment is made from natural scallop shells. However, the present invention is not limited to hydroxyapatite derived from scallop shells, and it is also possible to use natural hydroxyapatite derived from living organisms, such as coral, fish scales and bones, mammalian bones such as cow bones, seaweed, egg shells, sea urchin shells, corbicula shells, oyster shells, pearls, dolomite, and weathered shells (Pleurotus fasciatus). It is also possible to use hydroxyapatite derived from shells used as pearl mother bodies. In this case, shells such as pearl oysters and black lipped pearl oysters are suitable.

[0180] Unlike mineral-derived hydroxyapatite, natural biological hydroxyapatite is said to have high biocompatibility because it contains natural trace minerals such as magnesium and sodium in the apatite crystal structure. Furthermore, by adding deep sea water, it is possible to additionally replenish natural ionized minerals other than calcium.

[0181] In addition, in this embodiment, weathered shells (Camionisiki shells), fossil coral powder, and dolomite can be used instead of hydroxyapatite. Although weathered shells (Camionisiki shells), fossil coral powder, and dolomite are not hydroxyapatite, they contain high-quality minerals other than calcium, and are expected to have a synergistic effect with calcium supplementation on teeth and bones.

[0182] [Example of verification using immune cells between Bioapatite (registered trademark: derived from natural organisms and low crystallinity) and KT-11 lactic acid bacteria] The present inventors performed an evaluation test of the interleukin 12 (IL-12) production induction ability using immune cells. "IL-12" is a cytokine produced by immune cells that induces immune activation and anti-allergic action. It is also known that "IL-12" activates "natural killer cells" that control immune cells and enhances immunity. Furthermore, "IL-12" activates "Th1 cells" that induce anti-allergic action, and is useful for alleviating allergic symptoms. In this test, macrophage cells (immune cells) were used as cells. Macrophage cells were dispensed into a microplate, samples were added, and the cells were cultured, and the amount of IL-12 produced was measured by ELISA. As a result, as shown in Figure 6, when bioapatite and KT-11 lactic acid bacteria were used in combination (1% or 10% KT-11 lactic acid bacteria were added to 100% hydroxyapatite), "significant improvement in IL-12 production" was confirmed compared to when each was used alone. In addition, it was confirmed that bioapatite alone can produce IL-12.

[0183] [Anti-aging effect verification example using nematodes (in the case of granules)] The present inventors focused on the effect of the ionized apatite according to this embodiment on the average lifespan of nematodes, and conducted a study based on the results of a test conducted by a third party. In other words, it is said that by ingesting foods containing antioxidant components, the decline in the body's ability to remove active oxygen that occurs with aging can be suppressed, and lifestyle-related diseases caused by aging can be prevented, but the functional evaluation of foods is mainly performed "in vitro". In order to demonstrate the effect in the living body, "in vivo" biological tests are required. In this "in vivo" test, a type of nematode called "Caenorhabditis elegans" is an organism that can replace mice, etc. in this "in vivo" test. This "C. elegans" is a non-parasitic nematode that has a lifespan of about 3 weeks and lives in soil of about 1 mm. All of its genes have been elucidated, and it is used in tests around the world as a model organism for experiments to analyze various life phenomena.

[0184] The test method in this embodiment is as follows. That is, the bait E. coli OP-50 strain was suspended in the "S-medium" shown in the table below to prepare a culture solution for nematodes (fer-15 mutant strain). The nematodes were cultured (20°C, 100 rpm) using this culture solution until they became adults. After that, the nematodes were collected and washed in the "S-buffer" shown in the table below, and the bodies of the nematodes were dissolved using a NaOH solution and Haiter (registered trademark), and eggs were collected from the bodies. The collected eggs were cultured overnight at 20°C and hatched. The hatched L1 larvae were synchronously cultured at 26.5°C using heat-treated E. coli OP-50 strain (dead bacteria) as bait using a 24-hole microplate. At this time, the number of nematodes per well of the microplate was adjusted to about 20. The stock solution was prepared by dissolving ionized apatite (granules) in pure water at a concentration of 100 mg / mL. On the fifth day after egg collection, the stock solution was sterilized by filtration using a 0.22 μm filter, and added to each well of the microplate so that the final concentration in the culture solution was 10, 100, or 1000 times the stock solution. After that, the survival rate of the nematodes was checked under a microscope every few days, and the number of survivors on the fifth day was set to 100%, and a survival rate curve was drawn using the Kaplan Meier method, and the p-value was calculated using the log-rank test. The average lifespan was also calculated from the number of days of survival. The results are as follows: [Table 45] [Table 46] [Table 47]

[0185] The ionized apatite (granules) according to this embodiment showed a statistically significant extension of lifespan compared to the control in all treatment groups (p value < 0.05). In particular, when diluted 10 times, the lifespan was extended by 16.6% compared to the control, and even when diluted 1000 times, a statistically significant extension of the average lifespan was confirmed, indicating that a high anti-aging effect can be expected in practical terms even when diluted.

[0186] [Anti-aging effect verification example using nematodes (in the case of powder)] The present inventors focused on the effect of the ionized apatite according to this embodiment on the average lifespan of nematodes, and conducted a study based on the results of a test conducted by a third party. The test method was as follows. That is, the fertilized E. coli OP-50 strain was suspended in "S-medium" to prepare a culture solution for nematodes (fer-15 mutant strain). The culture solution was used to culture the nematodes until they became adults (20°C, 100 rpm). After that, the nematodes were collected and washed in "S-buffer", and the bodies of the nematodes were dissolved using a NaOH solution and Haiter (registered trademark), and eggs were collected from the bodies. The collected eggs were cultured overnight at 20°C and hatched. The hatched L1 larvae were cultured synchronously at 26.5°C using a 24-hole microplate and fed with heat-treated E. coli OP-50 strain (dead bacteria). At this time, the number of nematodes per well of the microplate was adjusted to about 20. The stock solution was prepared by dissolving ionized apatite (powder) in pure water at a concentration of 100 mg / mL. On the fifth day after egg collection, the stock solution was sterilized by filtration using a 0.22 μm filter, and added to each well of the microplate so that the final concentration in the culture solution was 10, 100, or 1000 times the stock solution. After that, the survival rate of the nematodes was checked under a microscope every few days, and the number of survivors on the fifth day was set to 100%, and a survival rate curve was drawn using the Kaplan Meier method, and the p-value was calculated using the log-rank test. The average lifespan was also calculated from the number of days of survival. The results are as follows: [Table 48]

[0187] The ionized apatite (powder) according to this embodiment showed a statistically significant extension of lifespan compared to the control in all treatments (p value < 0.05). In particular, the extension effect was highest at 1000 times dilution, which was even greater than the 10 and 100 times dilutions, and the lifespan was extended by 15.86% compared to the control, a notable figure. This shows that a high anti-aging effect can be expected in practical terms even when diluted.

[0188] Further, a comparative example will be described. Mineral-derived hydroxyapatite is used in many oral care products. The present inventors have investigated the "anti-aging effect" of mineral-derived hydroxyapatite based on the results of a test conducted by a third party. The test method is as follows. That is, Escherichia coli OP-50 strain, which serves as food, was suspended in "S-medium" to form a culture solution for nematodes (fer-15 mutant strain). The nematodes were cultured (20°C, 100 rpm) using this culture solution until they became adults. After that, the nematodes were collected and washed in "S-buffer", and the bodies of the nematodes were dissolved using NaOH solution and Haiter (registered trademark), and eggs were collected from the bodies. The collected eggs were cultured overnight at 20°C and hatched. The hatched L1 larvae were synchronously cultured at 26.5°C using a 24-hole microplate and fed with heat-treated Escherichia coli OP-50 strain (dead cells). At this time, the number of nematodes was adjusted so that there were about 20 nematodes per well of the microplate. Mineral-derived hydroxyapatite was dissolved in pure water at a concentration of 100 mg / mL to prepare the stock solution. On the fifth day after egg collection, the stock solution was filtered and sterilized with a 0.22 μm filter, and added to each well of the microplate so that the final concentration in the culture solution was 10, 100, or 1000 times the stock solution. After that, the survival rate of the nematodes was checked under a microscope every few days, and the number of survivors on the fifth day was set to 100%, and a survival rate curve was drawn using the "Kaplan Meier method" and the p-value was calculated using the log-rank test. The average lifespan was also calculated from the number of days of survival. The results are as follows. [Table 49] [Table 50] [Table 51]

[0189] Thus, mineral-derived hydroxyapatite was found to have a shorter average lifespan than the control in all treatment groups. In particular, mineral-derived hydroxyapatite at a 100-fold dilution showed a statistically significant shortening of the average lifespan compared to the control (p value < 0.05).

[0190] [Verification example of ionization rate] The inventors have verified the ionization rate of the ionized apatite according to this embodiment. The specimen was the ionized apatite according to this embodiment, and the specimens were scallop shell-derived hydroxyapatite and a dispersion liquid in which the scallop shell-derived hydroxyapatite was crushed and turned into nanopowder as Comparative Example 1. 50 mL of water was added to 5 g of each specimen, and the specimens were shaken for 30 minutes, and the phosphorus and calcium contents of the liquid filtered through a 0.45 μm membrane filter were measured by ICP emission spectrometry. However, the contents were calculated from the specimen weight. In addition, the pH was measured by the glass electrode method for a 1% aqueous solution. The measurement results are shown in the following table.

[0191] [Table 52] [Consideration] As a result of the above verification, the ionized apatite according to this embodiment was found to have values ​​that greatly exceed those of the original materials "hydroxyapatite derived from scallop shells" and "nano dispersion liquid of hydroxyapatite derived from scallop shells". In other words, it was found that the ionized apatite, which has a maximum hydroxyapatite content of 80%, has an ionization rate that is significantly higher than that of the original material, 100% hydroxyapatite derived from scallop shells. Furthermore, it was found that the ionized apatite has an ionization rate that is significantly higher than that of "a dispersion liquid in which hydroxyapatite derived from scallop shells is crushed to a <nano level> at a cost of several tens of thousands of yen per kg".

[0192] In order to prevent tooth demineralization from progressing to caries, it is known that it is important to use sugar alcohols such as xylitol, which do not produce acids that cause demineralization, as a substitute for sugar, and to supply calcium that is easily dissolved in saliva. Human saliva has sufficient phosphate ions necessary for remineralization, but not sufficient calcium ions. In other words, the phosphate ion concentration is sufficient, but the calcium ion concentration is less than half that amount. As described above, the ionized apatite according to this embodiment can supply a large amount of calcium ions together with phosphate ions, so that it is possible to dissolve sufficient calcium ions in saliva to replenish the minerals lost in the early stages of "dental caries". Hydroxyapatite supplies calcium ions and phosphate ions to teeth, and therefore promotes tooth remineralization. In addition, since calcium ions are relatively insufficient compared to phosphate ions in saliva, ionized apatite that can supply a large amount of calcium ions can be expected to have even more useful effects.

[0193] [Silk Hydrogen Pearl Powder Blend] The reduced fermented lactic acid bacteria according to this embodiment may contain silk hydrogen pearl powder in which hydrogen is carried by "silk powder impregnated with deep sea water" and "hydroxyapatite derived from pearl oysters". This makes it possible to synergistically obtain the benefits of silk, hydrogen, and hydroxyapatite derived from pearl oysters. In this embodiment, the explanation was given using "hydroxyapatite derived from pearl oysters". Pearl oysters grow in layers in a mille-feuille shape, and hydroxyapatite made from pearl oysters has the lightest bulk density of all bioapatites, making it suitable for carrying hydrogen. It is also expected that it will have the highest affinity with teeth at a lower crystallinity when used for oral care. In this way, by combining "silk powder impregnated with deep sea water" and "hydroxyapatite derived from pearl oysters", it is possible to obtain a "hybrid" hydrogen raw material (for food use, acidic) that generates both hydrogen gas and hydrogen ions. This combination has an oxidation-reduction potential of "-270 mV (25°C)" and a pH of "5.4 (22°C)". However, the present invention is not limited to these, and the redox potential can be further lowered, or made alkaline, and it is also possible to use "hydroxyapatite" that is not "derived from pearl oysters." Silk hydrogen pearl powder is a material that can simultaneously generate both hydrogen gas and hydrogen ions (electron-carrying hydrogen ions, also called negative hydrogen ions). The amount of hydrogen gas generated can be measured by gas chromatography testing at the Japan Food Analysis Center, and the amount of hydrogen ions generated can be measured by reacting "NAD+ reagent" with silk hydrogen pearl powder and measuring the number of NADH generated.

[0194] The inventors measured the amount of hydrogen generated from "Silk Hydrogen Pearl Powder" by gas chromatography. The test method is as follows. That is, after crushing the specimen (Silk Hydrogen Pearl Powder) in a mortar, 0.2g and 0.1g were taken and added to a 125mL vial. 25mL of purified water was added thereto, the vial was quickly capped, and ultrasonic extraction was performed for 30 minutes. After leaving it at room temperature for 48 hours or more, 0.5mL of headspace gas in the vial was injected into a gas chromatograph to measure hydrogen. As a result, "12mL" of hydrogen gas was detected per 1g of Silk Hydrogen Pearl Powder. Normally, when hydrogen gas is dissolved in water, it is said that the limit is a dissolved hydrogen concentration of 1.6ppm (a state in which 1.6mg of hydrogen is dissolved in 1L of water) at room temperature and normal pressure (15-25℃, 1 atm) (saturated hydrogen water), and 0.018mL of hydrogen gas is dissolved in 1mL of water, that is, 1.8mL of hydrogen gas is dissolved in 100mL of water. In contrast, 1g of Silk Hydrogen Pearl Powder provides the same amount of hydrogen gas as drinking six cups of 100mL (one glass) of saturated hydrogen water. Furthermore, the electron-charged hydrogen ions are generated through a synergistic effect with the pearl oyster-derived hydroxyapatite, resulting in a sustained effect on the amount of dissolved hydrogen, which is normally said to be "halved every three hours."

[0195] The inventors also measured the amount of hydrogen ions in the silk hydrogen pearl powder. The reagents used were "reduced NADH standard solution: manufactured by Wako Pure Chemical Industries, Ltd.", "potassium pyrophosphate buffer (F-kit)" and "NAD tablets (F-kit)". The measurement principle is "NAD + +H + +2e - →NADH", and the NADH produced by the generation of hydrogen is measured to calculate the amount of hydrogen. The test method is to first add 10mL of distilled water to 1g of reagent and stir, leave to stand for 1 hour, and then analyze the supernatant. The NADH molar concentration is calculated and this is taken to be the amount of hydrogen. The results are as shown in the following table. [Table 53]

[0196] The inventors measured the amount of dissolved hydrogen in a 1% aqueous solution of 0.5 g of Silk Hydrogen Pearl Powder dissolved and stirred in 50 mL of Yokohama City tap water using the Trustrex ENH-1000. As a result, the hydrogen content was 1035 ppb 10 minutes after addition, but 1106 ppb was measured 12 hours after addition. Furthermore, the hydrogen content was 1127 ppb 25 hours after addition, 1239 ppb 60 hours after addition, 1265 ppb 72 hours after addition, and 1286 ppb 96 hours after addition. Furthermore, the hydrogen content was 1274 ppb 108 hours after addition, and 1034 ppb 132 hours after addition.

[0197] Thus, even after 12 hours, the value did not decrease, but rather increased, which was a remarkable result. Even more surprisingly, the value increased even after 25, 60, 72, and 96 hours, confirming a high dissolved hydrogen amount of over 1000 ppb. A dissolved hydrogen amount of 1000 ppb is one of the benchmarks for high-concentration hydrogen water, but Silk Hydrogen Pearl Powder was able to achieve a dissolved hydrogen amount that exceeded this benchmark, and the high dissolved hydrogen amount of over 1000 ppb was maintained even after 108 and 132 hours, indicating that Silk Hydrogen Pearl Powder can maintain hydrogen dissolved in water for a long time. In this way, it can be said that the high dissolved hydrogen amount is achieved by the synergistic effect of the hybrid of hydrogen gas and hydrogen ions carrying electrons.

[0198] Drinking hydrogen water is expected to supply sufficient hydrogen to the intestines, where bad substances and bad bacteria react and where active oxygen is most produced in the body, and to the liver, which is the body's largest detoxifying organ and fights active oxygen (hydrogen absorbed from the intestinal tract is transported directly to the liver via the portal vein).In addition, according to an announcement by the Tokyo Metropolitan Institute of Gerontology, it has been confirmed that continued drinking of high-concentration hydrogen water inhibits vascular aging in mice, and as shown in Figure 13, hydrogen water is known to have various benefits.

[0199] The inventors also conducted a test on the effect of hydrogen bathing, in which hydrogen bathing powder was dissolved in warm water, on dry skin through Nagano Prefectural Nursing University. Dry skin is known to be one of the causes of skin troubles, caused by aging and ultraviolet rays. If the improvement effect of hydrogen bathing with hydrogen bathing additives is confirmed, it is expected to be one of the effective skin care methods. The test method was to anesthetize hairless rats before bathing and measure the moisture content of the keratin of the dry skin treatment and untreatment. After taking a picture with a digital camera, the rats were allowed to bathe for 15 minutes. This was performed 10 times during the 28-day test period. The hydrogen bathing was performed by adding 20g of hydrogen bathing additive to warm water at 42 degrees, and the initial dissolved hydrogen concentration was about 1000 ppb.

[0200] As shown in Figure 19, the results showed that the hydrogen water group increased the "stratum corneum moisture content" more than the warm water group in all baths. As shown in Figure 20, this effect was confirmed in the short term (six baths) and was even greater in the long term (12 baths). Furthermore, as shown in Figure 21, an effect was observed against moisture loss due to dry skin. Skin troubles are thought to be caused in part by oxidative stress, and it is believed that using substances with antioxidant properties is effective. The results of this test suggest that hydrogen's reducing effect worked against oxidative stress in the body, demonstrating the potential of hydrogen water baths as a form of skin care.

[0201] Since ancient times, silk has been valued as a high-end clothing material. Silk has often been used as a fiber, but in recent years, edible silk peptides have been developed using silk as a raw material. Silk peptides are functional peptides obtained by enzymatic decomposition of silk, and are known to have various physiological activities when their molecular weight is 300 to 5,000. It is said that consuming such silk peptides can restore the amount of moisture evaporation and elasticity of the skin, improve weight gain and fat gain, and prevent obesity.

[0202] According to publicly available information, in a "stress-induced skin damage test" using mice, silk peptide was confirmed to improve skin elasticity and moisture retention. In an "antioxidant test," silk peptide was added with peroxidase and the chemiluminescence inhibition rate was calculated, confirming that silk peptide has a strong antioxidant effect.

[0203] In addition, a test has been published in which a sheep red blood cell suspension was administered into the abdominal cavity of a mouse as an "immunostimulation test," and the spleen was removed four days later to prepare a spleen cell suspension, silk peptide was added to this suspension, cultured, and the amount of antibodies against sheep red blood cells produced in the culture medium was measured. As a result of this test, silk peptide was confirmed to have the effect of increasing antibody production, and it has been suggested that it exerts an immunostimulating effect.

[0204] In addition, in a cytokine production test using mice, it was confirmed that silk peptide specifically and strongly enhanced the production of IL-2, IL-4, and IFN-γ stimulated by Concanavalin A, which is expected to have an effect of activating lymphocyte cells in the body, preventing viral and bacterial infections, and having an antitumor effect. Furthermore, the cytokines IL-4 and IFN-γ are known to have the function of suppressing the differentiation of osteoclasts, which is expected to prevent osteoporosis.

[0205] Furthermore, silk peptide has been confirmed to have the effect of improving obesity, preventing and improving diabetes, and suppressing the occurrence of cancer caused by carcinogens, and has been confirmed to have no acute toxicity in acute toxicity tests and no subacute toxicity in subacute toxicity tests.

[0206] Regarding the structure of silk fibroin, for example, it is known that silk fibroin with a diameter of about 20 μm contains about 1000 fibrils, and each fibril contains about 900 microfibrils. Each microfibril contains about 350 fibroin molecules, and each fibroin molecule is composed of about 4000 amino acid bonds. Silk fibroin with such a porous structure has the function of absorbing dietary oils and fats and excreting them from the body, and is therefore attracting attention as a diet material.

[0207] On the other hand, the reactive oxygen species generated in the human body are said to cause skin aging such as age spots and wrinkles from the perspective of beauty, and also cause lifestyle-related diseases such as diabetes from the perspective of internal medicine. Such reactive oxygen species are generated simply by breathing, and are also generated by stress, excessive exercise, ultraviolet rays, radiation, overeating, and smoking. Such reactive oxygen species also play a role in breaking down, sterilizing, and removing bacteria and viruses, but if they increase too much in the body, they will attack the body and oxidize cells, DNA, mitochondria, and blood vessels. In particular, hydroxyl radicals, a typical example of bad reactive oxygen species, are known to have particularly strong oxidizing power and cause various diseases, aging, and fatigue.

[0208] In order to remove such active oxygen, people have started to drink "hydrogen water." It is said that hydrogen water filled in aluminum cans on the market contains 0.16mg-0.32mg (0.4ppm-0.8ppm) of hydrogen per 410mL at the time of shipment. However, hydrogen is a "light gas" that escapes and evaporates very easily, and there have been reported cases where the amount of dissolved hydrogen in aluminum cans has decreased significantly several months after production compared to the time of shipment. Recently, containers that prevent hydrogen from escaping have been developed, but it is said that it is important to drink the entire bottle within a few hours of opening it.

[0209] The reduced fermented lactic acid bacteria according to this embodiment may be configured to generate hydrogen by performing a special treatment using hydrogen and deep sea water. In this embodiment, for example, silk fibroin may be used as the hydrogen-carrying material, or silk powder made of silk peptide obtained by hydrolysis with an enzyme may be used. Various average molecular weights of the silk powder material have been proposed, but the present invention is not particularly limited thereto. In this embodiment, the raw material of the silk powder may be various forms, such as fibroin, peptide, and amino acid. In addition, silk powder may have a porous structure and has characteristics suitable for carrying hydrogen, but by mixing dextrin having a cyclic structure such as cyclodextrin, it becomes easier to carry hydrogen, and by adopting the manufacturing method of this embodiment, it is possible to carry a large amount of hydrogen that has not been seen in the past. Furthermore, the silk hydrogen pearl powder according to this embodiment can function as a "diet material" that adsorbs dietary fats and oils as silk fibroin, or it can exhibit functions such as antioxidant effects, skin beautifying effects, immune system improvement, and diet effects by using silk peptides and the like that are decomposed and absorbed as amino acids.

[0210] As described above, it is preferable to use hydroxyapatite (Bioapatite: registered trademark) that is naturally derived from a living organism and has low crystallinity, as the hydroxyapatite derived from pearl oysters. Hydroxyapatite is a component that accounts for approximately 97% of human tooth enamel and approximately 65% ​​of bone, and is composed of phosphoric acid and calcium. It is said to have high biocompatibility, is neutral to weakly alkaline, and is safe for the human body. Hydroxyapatite is composed of almost the same components as tooth enamel, and therefore has the function of remineralizing and repairing scratches on the surface of teeth caused by eating or brushing, and early stages of tooth decay caused by demineralization.

[0211] [Verification example of remineralization] The inventors conducted a test on remineralization. The teeth used in the test were "exfoliated human deciduous teeth," and the samples were (1) "hydroxyapatite containing only calcium raw materials" and (2) "Bioapatite (registered trademark)." The test method involved applying each sample to the surface of the "exfoliated human deciduous teeth," leaving it overnight, rinsing it off with tap water, brushing the surface 10 times with a toothbrush, and rinsing it again with tap water. After drying, the surface was observed with an electron microscope.

[0212] 7 to 9 are electron microscope photographs showing the results of tests on remineralization. All are magnified 1500 times. FIG. 7 shows the state before the test, and FIG. 8 shows the test results for "hydroxyapatite containing only calcium raw material." Also, FIG. 9 shows the test results for Bioapatite (registered trademark). As shown in FIG. 7, fine grooves were present on the surface before the test. In FIG. 8, the fine grooves remain, but in FIG. 9, the fine grooves have disappeared. This shows that Bioapatite (registered trademark) has filled in the fine grooves on the tooth surface. This confirmed that Bioapatite (registered trademark) is effective in remineralizing teeth.

[0213] Hydroxyapatite also has 10 calcium atoms in one molecule, and these calcium atoms have adsorption properties due to ion exchange. This ion exchange property makes it possible to hold large amounts of hydrogen and minerals contained in deep sea water. In addition, due to these adsorption properties, it is thought that when it is added to toothpaste or soap, it will function to adsorb pigments, bacteria, and lipid peroxides. It is also expected to adsorb dietary fats and oils when ingested.

[0214] Hydroxyapatite that is generally available is often synthesized using a mineral (apatite) as a raw material, but in this embodiment, it is a low-crystallinity apatite (bioapatite: registered trademark) made from natural pearl oyster (pearl shell). The present invention is not limited to hydroxyapatite derived from pearl oyster shells, and it is also possible to use hydroxyapatite derived from natural organisms, such as coral, fish scales and bones, mammalian bones such as cow bones, seaweed, egg shells, sea urchin shells, corbicula shells, oyster shells, scallop shells, dolomite, and weathered shells (Pleurotus fasciatus). It is also possible to use hydroxyapatite derived from shells used for pearl mother bodies. In this case, for example, shells such as black lipped pearl oysters are suitable. In addition, instead of hydroxyapatite derived from natural organisms, hydroxyapatite derived from minerals may be used.

[0215] Unlike mineral-derived hydroxyapatite, natural organism-derived hydroxyapatite is said to have high biocompatibility because it contains natural trace minerals such as magnesium and sodium in the apatite crystal structure. Furthermore, as described below, it is possible to support natural ionized minerals other than calcium by adding deep sea water. In addition, it is considered that not only natural organism-derived hydroxyapatite, but also mineral-derived hydroxyapatite can be supplemented with trace minerals by combining it with reduced deep sea water, thereby enhancing biocompatibility.

[0216] Hydroxyapatite derived from natural organisms includes hydroxyapatite derived from natural pearl oysters, hydroxyapatite derived from scallop shells, hydroxyapatite derived from coral, and hydroxyapatite derived from eggshells. The low-crystallized hydroxyapatite derived from natural organisms produced by Bioapatite (registered trademark)'s unique manufacturing method has all been certified as a quasi-drug ingredient standard and is a high-purity product with a high apatite content. For example, it has been reported that when eggshell-derived hydroxyapatite is used once a day for one month while brushing teeth, the eggshell-derived hydroxyapatite neatly fills in the scaly irregularities on the tooth surface, called "apatite prisms," and narrows the width of the cracks on the tooth surface.

[0217] In this embodiment, naturally occurring hydroxyapatite is used, but other materials capable of supplying a large amount of calcium ions and phosphate ions can also be used. For example, weathered shell calcium powder, fossil coral powder, or dolomite powder can supply a large amount of calcium ions, so it is possible to use any one of weathered shell calcium powder, fossil coral powder, and dolomite powder instead of naturally occurring hydroxyapatite.

[0218] Weathered shell calcium is calcium made from weathered shell fossils produced in the Yakumo region of Hokkaido. In the Yakumo region, the "nest shellfish" lived 15 to 20 million years ago, and there are sedimentary layers of these weathered shell fossils. These weathered shell fossils are calcium carbonate with a high purity of 95 to 97%, and are effective as a source of calcium ions. Fossil coral is found, for example, on Yonaguni Island in Okinawa Prefecture, and contains about 70 types of minerals such as calcium, magnesium, and iron. Dolomite is a biological ore that is formed when organisms such as coral are deposited on the seabed and become limestone, and then a part of the calcium is replaced with magnesium in seawater. Such dolomite contains calcium and magnesium in a 2:1 balance. In this embodiment, a large amount of calcium ions is supplied by using any one of weathered shell calcium powder, fossil coral powder, and dolomite powder.

[0219] In this embodiment, for example, "silk hydrogen pearl powder" is used as the hydrogen generating powder. When this "silk hydrogen pearl powder" is dissolved in water, "hydrogen molecules" and "electron-carrying hydrogen ions" are generated. Some of the hydrogen molecules escape as hydrogen gas, but in the aqueous solution, some of the hydrogen molecules ionize to form "positive hydrogen ions" and "negative hydrogen ions" as electron-carrying hydrogen ions. Positive hydrogen ions do not have the ability to release electrons, so they do not have reducing power, but negative hydrogen ions have the ability to release electrons, so they do have reducing power. The "hydrogen ions" learned in junior high and high schools are "H+ (positive hydrogen ions)" that are missing one electron. These negative hydrogen ions reduce active oxygen, and harmless water is generated. On the other hand, "antioxidants" such as vitamin C and polyphenols have the ability to remove active oxygen, but as a pro-oxidant, after removing active oxygen, they themselves remain as "oxides," which may cause the human body to become oxidized. On the other hand, hydrogen gas and negative hydrogen ions exert a reducing effect and, after combining with active oxygen, only produce harmless water, do not remain as an oxidant themselves, and have no pro-oxidant effect, making them the best antioxidants with no danger. In other words, hydrogen, as the smallest atom in the universe, is considered to be one of the best antioxidants in the universe, which is distributed throughout the human body, and is expected to recombine with antioxidants that have been oxidized and return them to their pre-oxidized state (reducing effect). Research into the use of such "electron-carrying hydrogen ions, also known as negative hydrogen," is also underway at the Tokyo Institute of Technology and Kyoto University. It is expected that hydrogen gas and electron-carrying hydrogen ions will work synergistically to exert antioxidant functions.

[0220] The inventors conducted a "human clinical trial" together with the "Japanese Society of Oxidative Therapy Medicine." A hybrid hydrogen supplement was created by mixing and filling "calcined coral calcium hydrogen powder" as a raw material for generating high concentrations of "electron-carrying hydrogen ions" with "marine mineral hydrogen powder" that utilizes deep seawater minerals as a raw material for generating high concentrations of hydrogen gas containing hydrogen ions. With this supplement, a statistically significant decrease was observed in the "average values ​​before and five days after ingestion" of the "8-OHdG creatinine ratio," a marker that indicates genetic damage in urine (oxidative stress in the body), just five days after starting to drink it. This revealed that this supplement can fully exert the "penetration power" of hydrogen gas and "electron-carrying hydrogen ions" in a short period of time.

[0221] In addition, the inventors measured the amount of dissolved hydrogen in the "calcined coral calcium hydrogen powder" over time in a test conducted at the request of the "Japan Hydrogen Water Promotion Association." As a result, the amount of dissolved hydrogen (redox method) was measured as 840 ppb after 10 minutes, 1,096 ppb after 2 hours, 1,085 ppb after 6 hours, and 1,045 ppb after 24 hours. In addition, at the Shiga Prefecture Northeast Industrial Technology Center, an investigation was conducted on the amount of hydrogen ion generation (inspection organization: Tres Bio Engineering LLC) using an ultraviolet-visible-near infrared spectrophotometer. The measurement principle is to measure the amount of hydrogen generated by the presence of hydrogen and calculate the amount of hydrogen. 10 mL of distilled water was added to 1 g of sample, stirred, and left to stand for 1 hour, and the supernatant was analyzed. As a result, it was found that the amount of hydrogen ions produced from 1 gram of powder was 3.55 x 10 to the power of 20 (3.55 sextillion), which is a level that can deliver 5.7 million hydrogen ions to each of the 60 trillion cells in the human body.

[0222] Furthermore, it was confirmed that hydrogen remained dissolved for a long time, and that a large amount of hydrogen ions was generated, so it can be expected that the "calcined coral calcium hydrogen powder" according to this embodiment will approach the intestines and the liver. It is said that about 90% of the active oxygen in the body is generated in the intestines, and the liver is the largest detoxification organ. The discovery of the possibility of approaching both organs is valuable data that shows the potential of hydrogen power. In addition, the acetaldehyde elimination ability was also investigated using an ultraviolet-visible-near infrared spectrophotometer at the Shiga Prefecture Northeast Industrial Technology Center (testing organization: Tres Bio Engineering LLC). The test was performed by adding 0.2 g of sample to 200 mL of shochu, stirring for 15 minutes, leaving it to stand, and then analyzing the supernatant. As a result, the aldehyde concentration was reduced from 10 mg / L to 1.9 mg / L, and an elimination rate of 81% was confirmed. The fact that it eliminates acetaldehyde, which is said to be the cause of hangovers, not only clarifies its effectiveness in preventing hangovers, but is also expected to lead to maintaining liver health.

[0223] In addition, the inventors requested a third-party organization (Designer Foods Co., Ltd.) to verify the hydroxyl radical scavenging function of the "calcined coral calcium hydrogen powder" according to this embodiment. The verification method is as follows. Since it is usually consumed at 420 mg to 820 mg / 100 mL, the calcined coral calcium hydrogen powder was suspended in distilled water (32.8 mg / mL) and left to stand for about 1 hour to make a stock solution. In the measurement, (1) a 10-fold diluted solution of the stock solution and (2) a stock solution were prepared and finally added to the active oxygen production system to obtain a 40-fold and 4-fold diluted solution (concentration for drinking), respectively. Regarding the "hydroxyl radical production method," hydrogen peroxide was generated by irradiating it with ultraviolet light.

[0224] Next, the prepared sample was collected in an ESR flat cell, and ESR measurements were performed under the following measurement conditions. Center Field: 335mT Modulation Width: 100μT Sweep Width: ±5.0mT Time Constant: 0.1sec Sweep Time: 1 min Gain: 50

[0225] Next, DMPO was used as a hydroxyl radical scavenger, and the hydroxyl radical scavenging activity of the samples was measured according to the following protocol. Distilled water (control), solution (1) or (2): 50 μL 5.7M DMPO: 20μL 2.5 mM H2O2: 130 μL After irradiating with ultraviolet light for 30 seconds, the ESR was measured.

[0226] In Fig. 22, the signal intensity of the obtained ESR spin adduct surrounded by a square was evaluated as "% of control" with the control being set as 100%. The signal intensity of the spin adduct was evaluated as "% of control" with the external standard sample Mn 2+ The analysis was performed based on the ratio of the ESR signal (left end of FIG. 22) to the ESR signal. Then, statistical processing was performed using the individual data. The number of data for each condition was 2 and 3, and statistical processing was performed using Tukey's analysis of variance, with a significance level of 5% or less.

[0227] The results showing the hydroxyl radical scavenging activity are shown in Figures 22 to 24. That is, compared to the control (a in Figure 22), 10.1% was eliminated for sample (1) 0.82 mg / mL (b in Figure 22), and 72.1% was eliminated for sample (2) 8.2 mg / mL (c in Figure 22). Thus, significant hydroxyl radical scavenging was observed for both (p<0.05, Tukey multiple test).

[0228] It is also considered effective to mix the reduced fermentation lactic acid bacteria according to this embodiment with at least one of the above-mentioned "silk hydrogen pearl powder" and "calcined coral calcium hydrogen powder."

[0229] [Combination with other ingredients] (A) Curcumin and Pea Sprout Extract (Pea Sprout Extract) Curcumin is a yellow polyphenol compound found in turmeric, and pea sprout extract (pea sprout extract) is a water-soluble extract extracted from pea sprouts, and is also sold as a raw material under the name Anagain (registered trademark). By blending these with the reduced fermented lactic acid bacteria powder according to this embodiment, hair, skin, and nail beautifying effects are expected. In particular, by combining it with "silicon" and "hydrogen" as described above, synergistic effects such as strengthening tissues and increasing antioxidant activity are expected. When commercializing the product, it may be "colored" using natural pigments to enhance its appeal.

[0230] (B) Rose petal powder, grape seed powder, hibiscus powder, cranberry extract, pomegranate, mangosteen, cherry blossom extract, melon extract Rose petal powder, for example, contains a lot of polyphenols derived from rose petals, and has the function of preventing skin aging, suppressing the absorption of fat from meals, and suppressing blood sugar information after meals. It also has antioxidant activity that exceeds that of vitamin C. Grape seed powder, hibiscus powder, and cranberry extract also contain a lot of vitamin C and polyphenols, have antioxidant activity, and have the effect of preventing periodontal disease and gingivitis, anti-calculus action, and cardioprotection. Pomegranate, mangosteen, and cherry blossom extracts also have high glycation inhibition and oxidation inhibition functions, so they may be added. Melon extract is an antioxidant material that induces three antioxidant enzymes in the body (SOD, glutathione peroxidase, and catalase) and removes excessively generated active oxygen, and is expected to enhance the skin's defense ability against ultraviolet rays, so it may be added for whitening care purposes. Furthermore, by combining these with citric acid and hydrogen, it is possible to further enhance the antioxidant and anti-glycation effects. In addition, when commercializing the product, it may be "colored" with natural pigments to enhance its marketability. It may also be blended with a "composite material of citrus and rosemary" as an ingredient that is expected to enhance the skin's defense ability against ultraviolet rays.

[0231] (C) Barley lactic acid fermentation extract, passion flower extract Barley lactic acid fermentation extract is rich in GABA (gamma-aminobutyric acid), and by combining it with passion flower extract, which has the function of regulating biological rhythms, it is expected to have effects on improving brain function, such as stabilizing hormone balance, mental relaxation, and good sleep. In particular, by combining it with hydrogen, "the smallest molecule in the universe that can even pass through the brain barrier," a high synergistic effect is expected. In addition, when commercializing it, it is possible to increase the product's appeal by "coloring" it with natural pigments such as spirulina, gardenia, and butterfly pea.

[0232] (D) Fulvic acid, humic acid, deep ocean water minerals, Great Salt Lake minerals By combining fulvic acid, humic acid, deep ocean water minerals, Great Salt Lake minerals, iron with the silicon, hydrogen, citric acid, etc. mentioned above, it is expected that the efficiency of mineral absorption will improve, tissues will be strengthened, and antioxidant effects will increase when ingested by the human body. That is, fulvic acid forms complexes with many metals in nature, but when complexed with iron, it becomes fulvic acid iron, which accounts for a large part of the movement of iron to the ocean and there is a lot of evidence that it promotes the growth of plants (including phytoplankton) and livestock. In addition, fulvic acid is one of the organic acids found in forests and soil, but it is taken up by humans and animals through the food chain starting from phytoplankton and plays a role in promoting the circulation of nutrients such as minerals that are transported into the body (ion exchange). The Great Salt Lake is a saltwater lake in the northern part of Utah, USA.

[0233] In recent years, a phenomenon called "isoyake" (iron bleaching) has been occurring along the coast of the Sea of ​​Japan in western Hokkaido, where the surface of rocks on the seabed turns pure white. When this phenomenon occurs, the amount of seaweed and phytoplankton at the bottom of the food chain shown in Figure 11 decreases, and as a result, the coastal fish that feed on them disappear, causing serious effects on the fishing industry. One of the causes is said to be the relationship between forest degradation and iron fulvic acid. Nitrogen is essential for the growth of seaweed and phytoplankton in the sea, but this nitrogen absorption requires iron ions, which act as a catalyst. Seawater contains only trace amounts of iron ions, and a decrease in the supply of iron from the forest through rivers leads to a shortage of iron ions. When considering the iron ions that are carried to the sea through rivers, "iron fulvic acid" is the keyword. In forests, leaves and branches that have fallen to the ground are decomposed by microorganisms, and fulvic acid is produced, which combines with the iron in the humus to become "iron fulvic acid." If iron remains as an ion, it will come into contact with oxygen while being carried to the river and oxidize, turning into "iron particles." However, as shown in Figure 12, iron ions that bind with fulvic acid in the forest travel down the river as "fulvic acid iron" and reach the sea "as iron ions." Iron fulvic acid plays an important role in the growth of phytoplankton and seaweed through nitrogen absorption.

[0234] "Fulvic acid" and "humic acid" are called humic substances, and are produced by the decomposition of organic matter, especially plants, and play a role in replenishing minerals to plants. Both have the function of transporting minerals and amino acids with chelating power (grabbing power) and excreting excess minerals. Fulvic acid is soluble in acidic solutions and is considered to be very rare. Humic acid is soluble in alkaline solutions and is composed of dark, black melanin pigments. The reason why "humic acid" is black is mainly because plants that grew in the shallow waters of the ancient sea, such as wakame seaweed, nori seaweed, and tree leaves, settled on the seabed, were trapped in thick deposits such as volcanic ash, and decomposed into organic matter over hundreds of millions of years. Humic acid is characterized by containing minerals that were bound to organic matter, and does not deteriorate even when exposed to the surface of the earth.

[0235] For example, a demonstration test of tidal flat purification using fulvic acid iron silicon has been conducted to enable the regeneration of the Ariake Sea, and it is also known that fulvic acid functions as a carrier that carries iron and water-soluble silicon from forests and mountains, and that tidal flats in lakes and oceans can be regenerated by fulvic acid iron silicon. Research is being conducted on the use of fulvic acid in the treatment of lifestyle-related diseases, cell function recovery through its ability to decompose harmful substances, skin beauty, improvement of atopic dermatitis and allergies, vision recovery, hair growth, and immune system improvement, for agricultural, livestock, human drinking, and cosmetics applications. In addition, by adding "deep ocean water minerals, Great Salt Lake minerals, and lactic acid bacteria" that have been reduced and processed with hydrogen to fulvic acid and fermenting it (called reduced fermented fulvic acid), the minerals are reduced and ionized, and the body is expected to absorb them without oxidizing them. In addition, since the color of the product will be brown granules if it is only fulvic acid, in order to improve the appearance and enhance the product's appeal (functionality), functional ingredients such as dark pink "rose petal powder, hibiscus powder, cranberry extract" that are known for their diet effects, anti-glycation, antioxidant, and antibacterial activities may be added to the product, resulting in a pink or wine red drink when dissolved in water. In addition, "tomato-derived lycopene extract powder" may be added as a natural tomato lycopene extract that contains a certain percentage of lycopene together with natural compounds contained in tomatoes (tocopherol, phytoene, phytofluene, β-carotene, phospholipid, phytosterol, etc.), resulting in a red-orange drink when dissolved in water.

[0236] Generally, it is said that fulvic acid and humic acid react with chlorine to generate trihalomethanes, and in waterworks, there is a process of removing fulvic acid and humic acid using activated carbon. For this reason, raw material manufacturers of fulvic acid and humic acid may warn on their product labels that "there is a concern that trihalomethanes may be generated, so do not mix with tap water and drink." In this situation, the inventors confirmed through a research institute that the reduced fermented fulvic acid according to this embodiment does not generate trihalomethanes even when mixed with the chlorine concentration of tap water. This test was conducted to measure how much total trihalomethanes are generated when the "reduced fermented fulvic acid powder" according to this embodiment is mixed with tap water. The test method is as follows. First, since the chlorine concentration of tap water is about 0.4 ppm, in this test, sodium hypochlorite (commercially available bleach) was diluted and adjusted to a chlorine concentration of 0.5 ppm to prepare chlorine water. Next, 0.2g of the sample "reduced fermented fulvic acid powder" was dissolved in 200mL of the above chlorinated water, sealed, and various trihalomethanes were measured (outside analysis was outsourced) according to the measurement method specified in "JIS S3201 (test method for household water purifiers) 6·1". The results are shown in the table below. [Table 54] The water quality standard for tap water is "0.1 mg / L or less" for total trihalomethane, which is the sum of the above four substances, but the reduced fermented fulvic acid according to this embodiment is less than 0.001. This confirmed that the reduced fermented fulvic acid according to this embodiment does not generate trihalomethane even when mixed with the chlorine concentration of tap water.

[0237] The present inventors also conducted a comparative test between "reduced and fermented fulvic acid powder" and "fulvic acid powder" through a research institute. Here, "reduced and fermented fulvic acid powder" refers to fulvic acid powder containing "deep ocean water minerals or Great Salt Lake minerals" and "lactic acid bacteria" that have been reduced and processed with hydrogen, and "fulvic acid powder" refers to fulvic acid powder before reduction processing. The reduced and fermented fulvic acid powder according to this embodiment utilizes high-quality fulvic acid powder produced in Canada, but fulvic acid produced in Utah, USA, can also be used.

[0238] (D1) Example of verification of antioxidant properties (SOD-like activity) The "reduced fermented fulvic acid powder" and "fulvic acid powder" were dissolved in pure water at a concentration of 1 mg / mL and filtered through a 0.45 μm filter to prepare the stock solution. The stock solution and the solution diluted 10-fold and 100-fold with the dilution buffer included with the "SOD Assay Kit-WST (Dojin Chemicals)" were used to measure the activity of the "SOD Assay Kit-WST (Dojin Chemicals)" according to the operating procedure. The measurement method is as follows. That is, using a 96-well microplate, the superoxide generated by the enzyme reaction was colored with tetrazolium salt WST-1, and the absorbance at 540 nm was measured. The inhibition rate of superoxide generation when the sample was added was calculated from the change in absorbance, and was taken as the SOD-like activity. The results are shown in the following table, and the higher the SOD-like activity, the higher the antioxidant property. [Table 55]

[0239] Next, for comparison, the antioxidant properties of fulvic acids manufactured by other companies were measured. Of the samples provided, the fulvic acid powder manufactured by Company B and the fulvic acid extract manufactured by Company E were dissolved in pure water to prepare a concentration of 1 mg / mL, and filtered through a 0.45 μm filter to prepare the stock solution. The other samples were provided and filtered through a 0.45 μm filter to prepare the stock solution. The inhibition rate was calculated according to the prescribed method using the SOD Assay Kit-WST for the stock solution and the solution obtained by diluting the stock solution 100-fold and 200-fold with the buffer solution included with the SOD Assay Kit-WST (manufactured by Dojindo Chemical Industries, Ltd.). The results are as follows. [Table 56] In the above table, negative numbers are equivalent to 0.

[0240] In this way, the collaboration between fulvic acid and "deep ocean water minerals and Great Salt Lake minerals" that have been reduced and processed with hydrogen has been shown to enhance the power of reduced and fermented fulvic acid more than simple fulvic acid. In other words, "reduced and fermented fulvic acid powder" has a high inhibition rate in the original solution. In addition, even if it is diluted 10 times or 100 times, it has a sufficiently high inhibition rate, so it is expected to have a sufficient effect even if it is diluted, and it was found that a sufficiently high antioxidant property can be obtained from the viewpoint of practical use. In particular, it was found that the "reduced and fermented fulvic acid powder" according to this embodiment is outstandingly superior, even when compared with fulvic acid liquid (or powder / extract) made by other companies. As a result, it can be expected to have an antioxidative effect on the body caused by active oxygen.

[0241] (D2) Example of verification of α-glucosidase inhibitory activity The "reduced fermented fulvic acid powder" and "fulvic acid powder" were dissolved in pure water at a concentration of 1 mg / mL and filtered through a 0.45 μm filter to prepare the stock solution. The α-glucosidase inhibitory activity of the stock solution and the solution diluted 10-fold and 100-fold with pure water was measured. The measurement method is as follows. That is, the substrate solution was prepared by dissolving "7 mM p-nitrophenyl-α-D-glucopyranoside" in 50 mM phosphate buffer (pH 7.0), and the enzyme solution was prepared by dissolving α-glucosidase at 0.9 U / mL in 50 mM phosphate buffer (pH 7.0). 10 μL of the sample solution and 40 μL of the enzyme solution were placed in a microtube and incubated at 37 ° C for 5 minutes, after which 950 μL of the substrate solution was placed and incubated at 37 ° C for 15 minutes. Then, 1000 μL of 0.5 M Tirs solution was added to stop the reaction. The absorbance at 405 nm of p-nitrophenol decomposed and liberated by this reaction was measured. The inhibition rate of p-nitrophenol production when the sample was added was calculated from the decrease in absorbance from the value when no sample was added, and this was taken as the α-glucosidase inhibitory activity. The results are shown in the following table, and the higher the α-glucosidase inhibitory activity, the higher the effect of suppressing the rise in blood glucose level. [Table 57] In the above table, negative numbers are equivalent to 0.

[0242] Next, for comparison, the α-glucosidase inhibitory activity of fulvic acid from other companies was measured. The fulvic acid powder from Company B and the fulvic acid extract from Company E were dissolved in pure water, adjusted to 1 mg / mL, and filtered through a 0.45 μm filter to prepare the stock solution. The other samples were provided and filtered through a 0.45 μm filter to prepare the stock solution. The stock solution and the liquid obtained by diluting the stock solution 10 times and 100 times with pure water were used as the measurement samples. 7 mM p-nitrophenyl-α-D-glucopyranoside was dissolved in 50 mM phosphate buffer (pH 7.0) to prepare the substrate solution, and α-glucosidase was dissolved in 50 mM phosphate buffer (A) at 0.9 U / mL to prepare the enzyme solution. 10 μL of the measurement sample and 40 μL of the enzyme solution were placed in a microtube and incubated at 37 °C for 5 minutes, and then 950 μL of the substrate solution was added and incubated at 37 °C for 15 minutes. Then, 1000 μL of 0.5 M Tirs solution was added to stop the reaction, and the absorbance at 405 nm was measured. The α-glucosidase inhibitory activity upon addition of the sample was calculated from the change in absorbance, and was taken as the α-glucosidase inhibitory activity. The results are shown in the table below. [Table 58] In the above table, negative numbers are equivalent to 0.

[0243] In this way, it was shown that the power of reduced and fermented fulvic acid is enhanced more than that of simple fulvic acid by collaboration between fulvic acid and "deep sea water minerals and Great Salt Lake minerals" that have been reduced and processed with hydrogen. In other words, since it has high α-glucosidase inhibitory activity, it is thought that it can inhibit α-glucosidase activity in the body, inhibit sugar absorption, and avoid high blood sugar levels. If high blood sugar levels can be avoided, it will be possible to create a state in which early AGEs production reaction products are less likely to be produced. In particular, it was found that the "reduced and fermented fulvic acid powder" according to this embodiment is remarkably superior to fulvic acid liquids (or powders and extracts) made by other companies.

[0244] (D3) Verification example of tyrosinase inhibitory activity The "reduced fermented fulvic acid powder" and "fulvic acid powder" were dissolved in pure water at a concentration of 1 mg / mL and filtered through a 0.45 μm filter to prepare the stock solution. The stock solution and the solution diluted 10-fold and 100-fold with pure water were measured for tyrosinase inhibitory activity. The measurement method is as follows. That is, the enzyme solution was prepared by dissolving mushroom-derived tyrosinase at 300 units / mL in 0.1 M phosphate buffer (pH 6.7), and the substrate solution was prepared by dissolving L-DOPA at 1 mM in 0.1 M phosphate buffer (pH 6.7). The measurement method is as follows. That is, using a 96-well microplate, 25 μL of sample solution and 100 μL of enzyme solution were added and incubated at 37 ° C for 10 minutes, and then 125 μL of substrate solution was added and incubated at 37 ° C for 30 minutes. The absorbance at 490 nm was then measured. The melanin production inhibition rate when the sample was added was calculated from the change in absorbance, and was taken as the tyrosinase inhibitory activity. The results are shown in the following table. [Table 59] In the above table, negative numbers are equivalent to 0.

[0245] Next, for comparison, the tyrosinase inhibitory activity of fulvic acid from other companies was measured. Of the samples provided, fulvic acid powder from Company B and fulvic acid extract from Company E were dissolved in pure water, adjusted to 1 mg / mL, and filtered through a 0.45 μm filter to prepare the stock solution. For the other samples, the provided samples were filtered through a 0.45 μm filter to prepare the stock solution. The stock solution and the liquid obtained by diluting the stock solution 10 times and 100 times with pure water were used as the measurement samples. The enzyme solution was prepared by dissolving mushroom-derived tyrosinase at 300 units / mL in 0.1 M phosphate buffer (pH 6.7), and the substrate solution was prepared by dissolving L-DOPA at 1 mM in 0.1 M phosphate buffer (pH 6.7). 25 μL of the measurement sample and 100 μL of the enzyme solution were placed in a 96-well microplate and incubated at 37°C for 10 minutes. 125 μL of the substrate solution was added thereto, and after incubation at 37° C. for 30 minutes, the absorbance at 490 nm was measured. The tyrosinase inhibition rate when the sample was added was calculated from the change in absorbance, and this was taken as the tyrosinase inhibitory activity. The results are shown in the table below. [Table 60] In the above table, negative numbers are equivalent to 0. In this way, the collaboration between fulvic acid and "deep ocean water minerals and Great Salt Lake minerals" that have been reduced and processed with hydrogen has been shown to enhance the power of reduced and fermented fulvic acid more than simple fulvic acid. In other words, the "reduced and fermented fulvic acid powder" has a positive tyrosinase inhibitory activity in the original solution, but even when diluted 10 times, it showed a positive value, so it is expected to be effective even when diluted, and it was found that the tyrosinase inhibitory activity function can be obtained from the viewpoint of practicality. In particular, it was found that the "reduced and fermented fulvic acid powder" according to this embodiment is relatively superior compared to fulvic acid liquids (or powders and extracts) made by other companies. This makes it difficult for melanin to be synthesized, and is expected to prevent skin blemishes.

[0246] (D4) Example of verification of hyaluronidase inhibitory activity The "reduced fermented fulvic acid powder" and "fulvic acid powder" were dissolved in pure water at a concentration of 1 mg / mL and filtered through a 0.45 μm filter to prepare the stock solution. The stock solution and the solution diluted 10-fold and 100-fold with pure water were measured for hyaluronidase inhibitory activity. The measurement method is as follows. The enzyme solution was prepared by dissolving bovine hyaluronidase at 400 units / mL in 0.1 M acetate buffer (pH 4.0). The substrate solution was prepared by dissolving potassium hyaluronate at 1 mg / mL in acetate buffer. The enzyme activator was prepared by dissolving Compund48 / 80 at 0.1 mg / mL in acetate buffer. The color development solution was prepared by dissolving p-dimethylaminobenzaldehyde (p-DABA) at 0.1 g / mL in a hydrochloric acid / acetic acid mixture, and was diluted 10-fold with acetic acid just before use. The measurement was carried out by putting 12 μL of sample solution and 12 μL of enzyme solution into a microtube and incubating at 40°C for 20 minutes. 12 μL of enzyme activator was added thereto and incubated at 40°C for 20 minutes. 12 μL of substrate solution was added and incubated at 40°C for 40 minutes. Then, 12 μL of 0.4N NaOH aqueous solution was added to stop the reaction, and the mixture was immediately cooled on ice for 5 minutes, 12 μL of 0.8M borate buffer (pH 9.0) was added, boiled for 3 minutes, and then cooled on ice for another 10 minutes. 180 μL of color developing solution was added thereto, and after incubation at 40°C for 30 minutes, the absorbance at 585 nm was measured. When the activity of hyaluronidase is inhibited when the sample is added, N-acetylglucosamine, a decomposition product of hyaluronic acid, is reduced, and the absorbance by p-DABA is reduced. The amount of change in absorbance was calculated to be the hyaluronidase inhibitory activity. The results are as shown in the following table. [Table 61] In the above table, negative numbers are equivalent to 0.

[0247] Next, for comparison, the hyaluronidase inhibitory activity of fulvic acid from other companies was measured. Of the samples provided, the fulvic acid powder from Company B and the fulvic acid extract from Company E were dissolved in pure water, adjusted to 1 mg / mL, and filtered through a 0.45 μm filter to prepare the stock solution. The other samples were provided and filtered through a 0.45 μm filter to prepare the stock solution. The stock solutions and the liquids obtained by diluting the stock solutions 10 times and 100 times with pure water were used as the measurement samples. The enzyme solution was prepared by dissolving bovine hyaluronidase at 400 units / mL in 0.1 M acetate buffer (pH 4.0). The substrate solution was prepared by dissolving potassium hyaluronate at 1 mg / mL in acetate buffer (pH 4.0). The enzyme activator was prepared by dissolving Compund48 / 80 at 0.1 mg / mL in acetate buffer (pH 4.0). p-Dimethylaminobenzaldehyde (p-DABA) was dissolved in a mixture of hydrochloric acid and acetic acid at 0.1 g / mL to prepare a coloring solution, which was diluted 10-fold with acetic acid immediately before use. 12 μL of sample solution and 12 μL of enzyme solution were placed in a microtube and incubated at 40°C for 20 minutes. 12 μL of enzyme activator was added and incubated at 40°C for 20 minutes. 12 μL of substrate solution was added and incubated at 40°C for 40 minutes. Then, 12 μL of 0.4N NaOH aqueous solution was added to stop the reaction, and the mixture was immediately cooled on ice for 5 minutes, 12 μL of 0.8M borate buffer (pH 9.0) was added, boiled for 3 minutes, and cooled on ice for 10 minutes. 180 μL of coloring solution was added, incubated at 40°C for 30 minutes, and the absorbance at 585 nm was measured. The hyaluronidase inhibitory activity when the sample was added was calculated from the change in absorbance, and was taken as the hyaluronidase inhibitory activity. The results are shown in the table below. [Table 62] In the above table, negative numbers are equivalent to 0.

[0248] In this way, it was shown that the power of reduced fermented fulvic acid is enhanced more than that of simple fulvic acid by collaboration between fulvic acid and "deep ocean water minerals and Great Salt Lake minerals" reduced and processed by hydrogen. Hyaluronic acid is said to exist in large amounts between cells and protects cells with its water retention and buffering functions, but hyaluronidase is an "enzyme that breaks down hyaluronic acid" that increases due to aging and ultraviolet irritation, and if it is present in large amounts, it is difficult to maintain the moisture retention and viscoelasticity of the skin, which may cause wrinkles and sagging of the skin. The "reduced fermented fulvic acid powder" according to this embodiment has high hyaluronidase inhibitory activity in the original solution. In addition, even if diluted 10 times or 100 times, it has a sufficiently high hyaluronidase inhibitory activity, so it is expected to have a sufficient effect even when diluted, and it was found that a sufficiently high hyaluronidase inhibitory activity function can be obtained from the viewpoint of practicality. In particular, it was found that the "reduced and fermented fulvic acid powder" according to this embodiment is significantly superior to other companies' fulvic acid liquids (or powders and extracts). This makes it difficult for hyaluronic acid to break down, and is expected to maintain the skin's moisturizing properties and viscoelasticity, preventing wrinkles and sagging skin.

[0249] (D5) Verification example of collagenase inhibitory activity The "reduced fermented fulvic acid powder" and "fulvic acid powder" were dissolved in pure water at a concentration of 1 mg / mL and filtered through a 0.45 μm filter to prepare the stock solution. The collagenase inhibitory activity of the stock solution and the solution diluted 10-fold and 100-fold with pure water was measured. The measurement method is as follows. Pz-peptide was dissolved in 0.1 M Tris buffer (pH 7.1) at 0.39 mg / mL to prepare the substrate solution, and collagenase Type IV was dissolved in pure water at 0.1 mg / mL to prepare the enzyme solution. 25 μL of sample solution, 25 μL of enzyme solution, and 200 μL of substrate solution were placed in a glass test tube and incubated at 37°C for 30 minutes, after which 250 μL of 25 mM citric acid was added to stop the reaction. Then, 2500 μL of ethyl acetate was added and the mixture was extracted by shaking, followed by centrifugation (1500 rpm, 5 minutes), and the upper layer (ethyl acetate layer) was collected and the absorbance at 320 nm was measured. The collagenase inhibitory activity was calculated from the change in absorbance when the sample was added, and was taken as the collagenase inhibitory activity. The test results are shown in the table below. [Table 63]

[0250] Next, for comparison, the collagenase inhibitory activity of fulvic acid from other companies was measured. Of the samples provided, fulvic acid powder from Company B and fulvic acid extract from Company E were dissolved in pure water, adjusted to 1 mg / mL, and filtered through a 0.45 μm filter to prepare the stock solution. For the other samples, the provided samples were filtered through a 0.45 μm filter to prepare the stock solution. The stock solution and the liquid obtained by diluting the stock solution 10 times and 100 times with pure water were used as the measurement samples. Pz-peptide was dissolved in 0.1 M Tris buffer (pH 7.1) at 0.39 mg / mL to prepare the substrate solution, and collagenase Type IV was dissolved in pure water at 0.1 mg / mL to prepare the enzyme solution. 25 μL of sample solution, 25 μL of enzyme solution, and 200 μL of substrate solution were placed in a glass test tube and incubated at 37 °C for 30 minutes, after which 250 μL of 25 mM citric acid was added to stop the reaction. Then, 2500 μL of ethyl acetate was added, and the mixture was extracted by shaking, then centrifuged (1500 rpm, 5 minutes), and the upper layer (ethyl acetate layer) was collected and its absorbance at 320 nm was measured. The collagenase inhibitory activity upon addition of the sample was calculated from the change in absorbance, and was taken as the collagenase inhibitory activity. The results are shown in the table below. [Table 64] In the above table, negative numbers are equivalent to 0.

[0251] In this way, the collaboration between fulvic acid and "deep ocean water minerals and Great Salt Lake minerals" that have been reduced and processed with hydrogen has been shown to enhance the power of reduced and fermented fulvic acid more than simple fulvic acid. In other words, it has been revealed that "reduced and fermented fulvic acid powder" has the function of inhibiting collagenase, which breaks down collagen. In particular, it has been found that the "reduced and fermented fulvic acid powder" according to this embodiment is comparable in quality to fulvic acid liquids (or powders and extracts) made by other companies. As a result, it is expected that collagen in the skin will be less likely to be broken down by ingesting or applying "reduced and fermented fulvic acid powder".

[0252] (D6) Example of verification of elastase inhibitory activity The "reduced fermented fulvic acid powder" and "fulvic acid powder" were dissolved in pure water at a concentration of 1 mg / mL and filtered through a 0.45 μm filter to prepare the stock solution. The elastase inhibitory activity of the stock solution and the solution diluted 10-fold and 100-fold with pure water was measured. The measurement method is as follows. That is, STANA (N-Succinyl-Ala-Ala-Ala-p-nitroanilide) was dissolved at 1 mM in 50 mM Tris buffer (pH 8.8) to prepare the substrate solution, and porcine pancreatic elastase was dissolved at 0.5 units / mL in 50 mM Tris buffer (pH 8.8) to prepare the enzyme solution. 50 μL of sample solution, 50 μL of enzyme solution, and 100 μL of substrate solution were placed in a 96-well microplate, incubated at 37°C for 30 minutes, and then the absorbance at 405 nm was measured. The elastase inhibitory activity when the sample was added was calculated from the change in absorbance, and was taken as the elastase inhibitory activity. The test results are shown in the table below. [Table 65]

[0253] Next, for comparison, the elastase inhibitory activity of fulvic acid from other companies was measured. Of the samples provided, the fulvic acid powder from Company B and the fulvic acid extract from Company E were dissolved in pure water, adjusted to 1 mg / mL, and filtered through a 0.45 μm filter to prepare the stock solution. For the other samples, the provided samples were filtered through a 0.45 μm filter to prepare the stock solution. The stock solutions and solutions diluted 10 and 100 times with pure water were used as measurement samples. STANA (N-Succinyl-Ala-Ala-Ala-p-nitroanilide) was dissolved at 1 mM in 50 mM Tris buffer (pH 8.8) to prepare the substrate solution, and porcine pancreatic elastase was dissolved at 0.5 units / mL in 50 mM Tris buffer (pH 8.8) to prepare the enzyme solution. Using a 96-well microplate, 50 μL of the measurement sample, 50 μL of the enzyme solution, and 100 μL of the substrate solution were placed and incubated at 37°C for 30 minutes, after which the absorbance at 405 nm was measured. The elastase inhibitory activity upon addition of the sample was calculated from the change in absorbance, and this was taken as the elastase inhibitory activity. The results are shown in the following table. [Table 66] In the above table, negative numbers are equivalent to 0.

[0254] In this way, the collaboration between fulvic acid and "deep ocean water minerals and Great Salt Lake minerals" that have been reduced and processed with hydrogen has been shown to enhance the power of reduced and fermented fulvic acid more than simple fulvic acid. In other words, it has been revealed that "reduced and fermented fulvic acid powder" has the function of inhibiting elastase, which breaks down elastin. In particular, it has been found that the "reduced and fermented fulvic acid powder" according to this embodiment is remarkably superior, even when compared with fulvic acid liquids (or powders and extracts) made by other companies. As a result, it is expected that by ingesting or applying "reduced and fermented fulvic acid powder", elastin in the skin will be less likely to be broken down.

[0255] (D7) Example of verification of anti-aging effect using nematodes The present inventors focused on the effect of the reduced fermented fulvic acid powder according to this embodiment on the average life span of nematodes, and conducted a study based on the results of a test conducted by a third party. The test method in this embodiment is as follows. That is, the E. coli OP-50 strain, which serves as food, was suspended in the "S-medium" shown in the table below to form a culture solution for nematodes (fer-15 mutant strain). The nematodes were cultured (20°C, 100 rpm) using this culture solution until they became adults. After that, the nematodes were collected and washed in the "S-buffer" shown in the table below, and the bodies of the nematodes were dissolved using a NaOH solution and Haiter (registered trademark), and eggs were collected from the bodies. The collected eggs were cultured overnight at 20°C and hatched. The hatched L1 larvae were synchronously cultured at 26.5°C using a 24-hole microplate and fed with heat-treated E. coli OP-50 strain (dead bacteria). At this time, the number of nematodes per hole in the microplate was adjusted to about 20. The sample stock solution was prepared by dissolving reduced fermented fulvic acid powder in pure water at a concentration of 10 mg / mL. After collecting the eggs, the stock solution was filtered and sterilized with a 0.22 μm filter on the fourth day, and the sample stock solution was added to each well of the microplate so that the final concentration in the culture solution was 10 times diluted to "1 mg / mL", 100 times diluted to "0.1 mg / mL", and 1000 times diluted to "0.01 mg / mL". After that, the survival rate of the nematodes was checked under a microscope every few days, and the number of survivors on the fourth day was set to 100%, and a survival rate curve was drawn using the "Kaplan Meier method", and the p value was calculated using the log-rank test. The average lifespan was also calculated from the number of days of survival. The results are as follows. [Table 67] [Table 68] [Table 69] Thus, there was a tendency for the average life span to be slightly longer in the 1000-fold and 100-fold diluted treatments compared to the control. In addition, a significant difference was observed in the 100-fold diluted treatment compared to the control at p-value < 0.1.

[0256] (E) Combination with red orange extract, pineapple enzyme, papaya enzyme, hydrogen, and deep sea water Red orange extract refers to a powdered extract extracted from three types of red oranges (Moro, Zanghinello, and Tarocco) that are endemic to Sicily, Italy. When combined with hydrogen and deep sea water, it is expected to have anti-glycation, antioxidant, UV protection, and fat burning effects. In addition, by adding proteolytic enzymes such as pineapple enzymes and papaya enzymes, the ability to break down proteins into amino acids is enhanced, and it is expected to have an effect of "improving the intestinal environment by preventing protein spoilage" and dieting effects. When commercializing the product, it is acceptable to "color" it with natural pigments to increase its appeal.

[0257] In addition, by combining various proteins such as soy protein and pea-derived protein with proteolytic enzymes such as pineapple enzymes, papaya enzymes, and green papaya enzymes, the conversion and absorption efficiency into amino acids can be increased, and further combining functional edible charcoal, reduced fermented lactic acid bacteria, hydrogen, deep sea water, etc., an excellent collaboration can be expected. In addition to efficient intake of protein and amino acids during exercise and dieting, it can also adsorb and excrete bad substances, bad bacteria, active oxygen, AGEs (advanced glycation end products), etc. in the intestines, promoting intestinal cleansing.

[0258] (F) Functional coconut shell activated carbon "Functional coconut shell activated carbon granules (dispersion processing)" (hereinafter referred to as "coconut shell activated carbon") has excellent water dispersibility and is a porous preparation, so it is expected to be functional as a coating agent for bacteria, etc. The inventors immersed three types of bacteria in a dilute hydrochloric acid solution that resembles gastric acid, measured the number of bacteria in the bacteria mixed with coconut shell activated carbon and those not mixed, and measured the difference in coating power. The test method is as follows. That is, starting with colonies of various bacteria, one colony was suspended in sterile physiological saline to make an original bacterial solution. The original bacterial solution was divided into a group with charcoal added and a group without it, and dilute hydrochloric acid was added to each group and left to stand. C. butyruicum (butyric acid bacteria) and B. pseudolongum (bifidobacteria) were applied to GAM medium, and L. acidophilus (lactic acid bacteria) was applied to LGB medium, and then cultured anaerobically and the number of bacteria was measured. The results are as shown in the following table.

[0259] [Table 70]

[0260] Thus, in each test group, the number of viable bacteria was detected in the coconut shell activated carbon administration group more than in the non-administration group. This suggests that the coconut shell activated carbon, which has dispersibility, can be suspended in water to function as a carrier that maintains the number of viable bacteria even in a simulated gastric acid environment. From this, the reduced fermentation lactic acid bacteria according to this embodiment may be viable bacteria, and in particular, when coconut shell activated carbon is contained, a synergistic effect can be expected with yogurt containing each viable bacteria of lactic acid bacteria, bifidobacteria, and butyric acid bacteria.

[0261] Next, we will explain the caffeine adsorption test of "functional coconut shell activated carbon powder" and "Kishu Bincho activated carbon powder". The test method is as follows. Caffeine was adjusted to 600 ppm, and 1 g each of "functional coconut shell activated carbon powder" and "Kishu Bincho activated carbon powder" was added to 100 mL of the 600 ppm caffeine solution, stirred with a magnetic stirrer for 3 minutes, and then filtered with filter paper (Advantech No. 2). Next, the concentration was measured using a liquid chromatograph. The results are shown in the following table.

[0262] [Table 71] Thus, both "functional coconut shell activated carbon" and "Kishu Bincho activated carbon" showed high caffeine adsorption rates. Caffeine is said to have the effect of revitalizing the body if consumed in moderate amounts, but drinking large amounts has side effects, so coffee from which the caffeine has been removed is called decaffeinated and has been attracting attention. Note that the present invention is not limited to functional coconut shell activated carbon, and it may also be blended with "charcoal or activated carbon (hereinafter referred to as "functional edible charcoal")" that has high adsorption performance and is carbonized using temperature, pressure, reduced water, etc. For example, functional edible charcoal such as "Ina Akamatsu Myotan (registered trademark)" and "Kamakura Keichiku Charcoal (registered trademark)" may also be blended.

[0263] Next, we will explain the acrylamide adsorption test of "functional coconut shell activated carbon powder" and "Ina Akamatsu Myo Charcoal Powder". The test method is as follows. 30 mg of acrylamide was dissolved in 200 mL of distilled water, and 1 g of each sample was added to 45 mL of the acrylamide aqueous solution. Next, after stirring for 30 minutes, centrifugation (8000 rpm: 10 minutes) was performed, and the supernatant was filtered through a filter and the concentration was measured using a TOC meter (SHIMADZU TOC-V csn (total organic carbon meter)). The results are as shown in the following table.

[0264] [Table 72] In this way, both "functional coconut shell activated carbon (powder)" and "Ina Akamatsu Myotan (powder)" showed high acrylamide adsorption rates. This makes it possible to remove a high percentage of acrylamide when mixed with many commercially available dark roasted coffees.

[0265] Next, we will explain the phenol adsorption test of "Kishu Binchotan Activated Carbon Granules (Dispersed)". The test method is as follows. Phenol was adjusted to 15 ppm, and 1 g of the sample (Kishu Binchotan Activated Carbon Granules (Dispersed)) was added to 100 mL of this, stirred with a magnetic stirrer for 10 minutes, filtered, and then analyzed. The results are shown in the following table. [Table 73] In this way, "Kishu Binchotan Activated Carbon Granules (Dispersed)" showed a high phenol adsorption rate of over 90%. Recently, it has been reported that one of the causes of diabetic kidney disease is "phenylsulfate" produced by a specific group of intestinal bacteria. Phenylsulfate is made from the amino acid "tyrosine" found in foods such as cheese and meat, which is converted into a substance called "phenol" by an enzyme possessed by a specific group of intestinal bacteria. It is then absorbed from the intestinal tract, reaches the liver, where it is converted into phenylsulfate and released into the blood. As shown above, "Kishu Binchotan Activated Carbon Granules (Dispersed)" showed a high phenol adsorption rate, so it is expected that phenol, the source of phenylsulfate produced in the human body, will be removed, which is expected to have a preventive effect on diabetic kidney disease.

[0266] Next, a bile acid adsorption test using functional coconut shell activated carbon will be described. The inventors confirmed the bile acid adsorption effect of functional coconut shell activated carbon. In a test conducted by a third-party organization using an enzyme method and a reagent, the functional coconut shell activated carbon had an adsorption rate of 97%. Bile acids are produced in the liver from bad cholesterol and play a role in further breaking down "oil emulsified by the digestive enzyme lipase" and absorbing it into the body. About 95% of bile acids are absorbed from the intestinal tract together with oil and returned to the liver for reuse (called enterohepatic circulation), so that new synthesis of bile acids in the liver using bad cholesterol does not proceed, resulting in a vicious cycle in which the concentration of bad cholesterol in the blood increases. In other words, it can be said that the situation is like old engine oil circulating in the body. In addition, Professor Mitsuhiro Watanabe of the Graduate School of Media and Governance at Keio University (also Professor at the Faculty of Environment and Information Studies, Professor at the Faculty of Medicine (concurrent)) and his colleagues, in collaboration with a research group at the École Polytechnique Fédérale de Lausanne in Switzerland (Professor Johan Auwerx), have elucidated the mechanism by which metabolic syndrome is improved by bile acid regulation. The results of this research have been published in PLos ONE and Nature Scientific Reports, respectively, and the paper reports that by eating bile acid-adsorbing materials such as konjac and excreting bile acids in the stool, the risk of diabetes and blood cholesterol levels can be reduced. It is said that if you suffer from both hyperglycemia and hypercholesterolemia, the risk of vascular diseases such as cerebral infarction and myocardial infarction increases by about five times. It is estimated that there are about 8.9 million diabetes patients in Japan, about 13.2 million prediabetes patients, and about 25 million hypercholesterolemia patients. Early treatment and intervention for diabetes and borderline hypercholesterolemia is an important future issue in the global medical field. Furthermore, with the spread of Western food in Japan, colon cancer has become a major cause of death, and is the leading cause of cancer death in women. Eating a Western diet that is high in oil causes a lot of bile acids to be secreted, and while about 95% of these are absorbed into the body along with the oil, the remaining 5% travels to the large intestine. These react with bad bacteria in the large intestine and turn into "secondary bile acids," which are powerful carcinogens, and are said to be the main cause of colon cancer.Functional edible charcoal, including functional coconut shell activated charcoal, adsorbs and excretes bile acids, while reduced fermentation lactic acid bacteria lead to the elimination of bad bacteria, which can be expected to have synergistic effects such as lowering bad cholesterol levels, reducing the risk of diabetes, and reducing the risk of colon cancer.

[0267] (G) Chlorogenic Acid It is also possible to blend chlorogenic acid contained in coffee with the reduced fermentation lactic acid bacteria according to this embodiment. The health benefits of coffee are obtained from the action of chlorogenic acid, which is known to be contained in large amounts in lightly roasted coffee beans. In addition, products containing chlorogenic acid for the purpose of improving high blood pressure are also being offered on the market. It is also known that chlorogenic acid has the function of "inhibiting the action of sugar-decomposing enzymes (digestive enzymes)," "suppressing the transfer of sugar from the digestive tract to the blood," and "promoting the production of the hormone GLP-1, which is provided from the digestive tract and suppresses the rise in blood sugar levels." However, many of the commercially available coffees that are widely used in the world are dark roasted to hide the differences in the origin of the coffee beans and the deterioration of their quality, and it has been reported that oxidation progresses, chlorogenic acid is greatly reduced, the sourness of citric acid is lost, while the carcinogenic substance acrylamide is produced and causes health problems. A certain amount of caffeine is also said to be effective in activating the body, but caution is required as excessive intake can cause side effects.

[0268] (H) Hydrocharcoal (a charcoal-containing composition combining at least one of activated charcoal or edible charcoal, inulin, kagome kelp powder and citric acid) It is also possible to blend the reduced fermented lactic acid bacteria according to this embodiment with "hydrocharcoal," a charcoal-containing composition that combines at least one of activated charcoal and edible charcoal, inulin, Gagome kelp powder, and citric acid. In particular, activated charcoal (functional edible charcoal including functional coconut shell activated charcoal) has the function of highly adsorbing oral bacteria, the function of highly adsorbing synthetic coloring agents "tar dyes," heavy metals, purines, AGEs, acrylamide, nicotine, and tar, the function of highly adsorbing malondialdehyde, a lipid peroxide, and dietary fats and oils, and the function of highly adsorbing trihalomethanes and residual chlorine. Furthermore, by suspending it in water, it has a protective and transporting function that delivers useful microorganisms such as lactic acid bacteria alive to the intestines, that is, a function as a carrier that maintains the number of live bacteria. In addition to Gagome kelp, seaweed-derived materials such as akamoku, mekabu, ganiashi, and seaweed dulse may also be used.

[0269] The hydrocharcoal according to this embodiment may contain inulin. Inulin is a type of polysaccharide made by plants of the Asteraceae family, and has been increasingly used in foods in recent years due to its excellent nutritional value. That is, inulin belongs to the saccharide group such as sugar and starch, but humans do not have the enzyme to break down inulin, so even if they ingest food containing inulin, it is hardly absorbed and is excreted from the body. Therefore, inulin is classified as a water-soluble dietary fiber, and is known to become fructooligosaccharides when fermented and broken down in the intestine. When inulin absorbs water in the intestine, it becomes gel-like and has the function of suppressing the absorption of carbohydrates ingested together. In addition, since inulin serves as food for good bacteria in the intestine, it has the effect of regulating the intestinal environment, and is often used in diet foods. By ingesting inulin, humans can expect to improve the intestinal environment and suppress the rise in blood sugar and cholesterol levels.

[0270] Furthermore, it is expected that Gagome kelp powder will promote hair growth when applied to the human body. In recent years, it has been scientifically proven that a component called "fucoidan" contained in seaweed contributes to hair growth. "Fucoidan" can be extracted from several different seaweeds, such as kelp, wakame, and mozuku, but it has been found that the fucoidan in Gagome kelp in particular has a high hair growth effect. Gagome kelp is a type of kelp that can only be harvested in a limited area of ​​Hokkaido, and the Gagome kelp fucoidan extracted from this kelp is said to have a much higher hair growth effect than the fucoidan that can be extracted from other seaweeds. More specifically, the coastal foreshore of Minamikayabe in Hakodate, where Gagome kelp is produced, is shallow, has high light intensity, and has water temperatures that are suitable for growing kelp due to the confluence of hot and cold currents. In addition, the coastal foreshore has acidic rocky terrain that is rich in silicon, nutrients from broadleaf forests, and the mineral iron fulvic acid that flows in from 30 rivers, large and small. These favorable conditions are said to be ideal for growing high-quality kelp.

[0271] Recent research has shown that there are three types of fucoidan in gagome kelp: "F-fucoidan," "U-fucoidan," and "G-fucoidan," and that "F-fucoidan" has a particularly strong hair growth effect. Gagome kelp fucoidan is said to increase the production of hair growth factors and promote the proliferation of hair matrix cells. This growth factor is a type of protein called "FGF-7," which is said to extend the growth phase of the human hair cycle and extend the hair growth period. Hair can grow only as much as the growth period is extended, making it possible to grow thicker and stronger hair. It also has the effect of transitioning hair roots that have entered the resting phase into the growth phase more quickly, further extending the hair growth period. Gagome kelp also has a high moisturizing effect unique to seaweed, and is said to retain moisture in the scalp and maintain a good scalp environment.

[0272] In addition to fucoidan, gagome kelp is known to contain iodine and silicon. Iodine is found in large amounts in seafloor sediments and is incorporated into seaweed. It is also known that silicon has a positive effect on healthy growth due to the above-mentioned growing environment of gagome kelp. Silicon is present in the human body's nails, hair, bones, cell membranes, etc., and has the function of strengthening tissues through their cores and protecting cells from oxidation and glycation. In addition, as will be described later, when hydrogen is absorbed, gagome kelp's "thickness" makes it difficult for hydrogen to escape.

[0273] Citric acid is a weak acid with three carboxyl groups, and is found in citrus fruits (mandarins, limes, lemons, grapefruit, etc.). It is also widely used as a food additive due to its sour taste. Citric acid is a component of the "citric acid cycle" in the body, and is widely used as a supplement, mainly for the purpose of energy production through the citric acid cycle. Citric acid also increases the solubility of calcium through its "chelating effect," which promotes calcium absorption from the small intestine.

[0274] Citric acid has various effects, and is known to have the main effects of fatigue recovery, skin beautifying effects, and hair growth. For example, "human hair" is weakly acidic, but shampoo is weakly alkaline, so washing your hair can disrupt the pH balance. After washing your hair with shampoo, your hair may feel squeaky, which means that your hair has become alkaline. Citric acid is useful for restoring this condition. By neutralizing hair that has become alkaline with citric acid, it is possible to restore the condition of your hair to its original state. By using citric acid as a rinse after shampooing, the condition of your hair can be restored, improving the feel, and also promoting blood circulation and softening the scalp. It is also said to be effective in preventing thinning hair and hair loss, and improving split ends and breakage.

[0275] The hydrocharcoal according to this embodiment contains deep sea water. Deep sea water is generally understood to be sea water at a depth of 200 m or more, and has the characteristics of cleanliness, abundance of inorganic nutrients, and low temperature stability compared to surface water. That is, deep sea water is not affected by river water polluted by human wastewater, so there is no contamination by chemicals, and since plankton does not grow because sunlight does not reach it, the amount of harmful bacteria is less than one thousandth of that of surface water. In addition, compared to surface water, it is rich in inorganic nutrients necessary for the growth of phytoplankton, and furthermore, it has the characteristics that the water temperature and contained components are less likely to change, and the water quality is stable.

[0276] In the hydrocharcoal according to this embodiment, the deep sea water is reduced by hydrogen. By performing the reduction treatment using hydrogen, hydrogen is absorbed, and when dissolved in water, the coconut shell activated carbon is quickly dispersed. In this embodiment, an example is shown in which the deep sea water is reduced by hydrogen and then mixed with coconut shell activated carbon, but the present invention is not limited to this, and it is also possible to perform the reduction treatment by hydrogen after the charcoal-containing composition is completed. In addition, the present invention does not necessarily require the deep sea water to be reduced by hydrogen. In addition, the deep sea water may be handled in a powdered state for convenience, but the present invention is not limited to the powder.

[0277] It is also possible to mix chlorogenic acid into hydrocharcoal, which contains inulin, functional edible charcoal, citric acid, gagome kelp, and hydrogen, and mix it with many commercially available dark roasted coffees for drinking. In this case, the functional edible charcoal adsorbs caffeine and acrylamide, while it is possible to replenish the chlorogenic acid and citric acid lost by dark roasting. As a result, it is possible to expect effective use for high blood pressure and hyperglycemia. When mixing chlorogenic acid, it is possible to use chlorogenic acid alone, but it is also possible to adopt a method of mixing green coffee bean extract, which is rich in chlorogenic acid. It is also possible to add "dried mango leaf extract" to the hydrocharcoal. In other words, charcoal adsorbs caffeine, and when mixed with coffee, it becomes decaffeinated, while it is possible to replenish the valuable antioxidant "chlorogenic acid" lost from dark roasted coffee by adding green coffee bean extract. Since some people dislike losing the stimulating effect of caffeine, adding dried mango leaf extract, which is a caffeine substitute and has a stimulating effect, will support decaffeinated coffee in its entirety and allow people to drink coffee in a healthy way. In addition, dried mango leaf extract has been confirmed to have the effect of improving mental energy and motor function.

[0278] (I) Synergistic effects of simultaneously ingesting reduced fermentation lactic acid bacteria, functional edible charcoal, and hydrogen First, the adsorption of methyl mercaptan by functional edible charcoal will be described. The inventors conducted an adsorption test of methyl mercaptan on (a) Ina Akamatsu Myo charcoal powder, (b) functional coconut shell activated charcoal powder, (c) Kamakura quartz bamboo charcoal powder, and (d) Kishu Bincho activated charcoal powder. The equipment used was a gas chromatograph mass spectrometer QP5050A manufactured by Shimadzu Corporation, and the reagent used was a methyl mercaptan standard solution 1 μg / μL manufactured by Wako Pure Chemical Industries, Ltd. The test method was as follows. That is, 0.1 g of the reagent was placed in a GCMS headspace vial, and 10 μL of the standard solution was injected along the inner wall surface of the vial. The vial was immediately sealed and left to stand for 20 minutes, after which methyl mercaptan was analyzed by GCMS. The results are shown in the following table. [Table 74] As described above, each functional edible charcoal showed a high adsorption rate for methyl mercaptan, which is the cause of bad odor. As a result, the reduced fermentation lactic acid bacteria showed elimination activity against Porphyromonas gingivalis, the causative bacterium of methyl mercaptan production, and the functional edible charcoal itself adsorbed methyl mercaptan, and therefore an excellent synergistic effect between the two can be expected in oral care applications.

[0279] Next, we will explain the AGE control effect of the collaboration between functional edible charcoal and reduced fermentation lactic acid bacteria. It is known that functional edible charcoal can adsorb AGEs produced in food. Figure 10 is a diagram explaining the mechanism of glycation. As shown in Figure 10, when blood sugar levels in the human body rise, the body heats up and AGEs are produced in the body. Reduced fermentation lactic acid bacteria suppress the production of AGEs in the body, and functional edible charcoal adsorbs AGEs in food. In this way, the complex functions of the collaboration between reduced fermentation lactic acid bacteria and functional edible charcoal make it possible to take comprehensive measures against the two major causes of glycation.

[0280] Next, we will explain the effect of controlling the two major aging factors, oxidation and glycation, through the collaboration of hydrogen, functional edible charcoal, and reduced fermentation lactic acid bacteria. It is said that about 90% of the active oxygen in the human body is generated in the intestines. The causes are bad substances such as food additives and heavy metals, and bad bacteria that increase with age. Hydrogen reacts with the active oxygen itself and is converted into safe and harmless water. It is also the smallest antioxidant in the universe with an atomic weight of 1, and is expected to spread to every corner of the body and prevent oxidation. However, since it is not possible to keep drinking hydrogen, it is important to take functional edible charcoal, oligosaccharides, water-soluble dietary fiber, and lactic acid bacteria at the same time as a fundamental approach, so that the functional edible charcoal adsorbs bad substances and the oligosaccharides, water-soluble dietary fiber, and lactic acid bacteria promote bowel movements. In addition, by adding the antibacterial properties of reduced fermented lactic acid bacteria, the triple action of hydrogen to eliminate active oxygen, functional edible charcoal to adsorb and remove harmful substances, and reduced fermented lactic acid bacteria to eliminate harmful bacteria makes it possible to take a total approach to the causes of oxidation. This makes it possible to address both "oxidation" and "glycation," which are considered to be the two major causes of aging.

[0281] (J) A combination of multiple types of oligosaccharides, KT-11 lactic acid bacteria, which "has a strong immune-stimulating effect and is expected to have a synergistic effect with complex lactic acid bacteria," green papaya extract, which "breaks down proteins, which are a factor in worsening the intestinal environment due to putrefaction, into amino acids," deep sea water, and hydrogen, which are expected to have a synergistic effect. These are expected to have a significant effect in promoting intestinal care. In addition, when commercializing the product, it is acceptable to "color" it with natural dyes to enhance its appeal. Here, the multiple oligosaccharides include galactooligosaccharides, xylooligosaccharides, fructooligosaccharides, lactofructose, and lactulose, which is a milk oligosaccharide. By using any of these in combination, each oligosaccharide will become food for the good bacteria, and it is expected that the advantages of each will create a synergistic effect. In addition, it has been reported that some of these oligosaccharides react with intestinal bacteria to increase the generation of hydrogen gas in the intestines, and it is expected that the hydrogen gas will be taken up from the intestinal tract into the blood, and the breath concentration of hydrogen gas will increase, which will have a beneficial effect on the body's antioxidant mechanism. By collaborating with deep seawater that has been subjected to reduction treatment to generate hydrogen gas and electron-charged hydrogen ions, it is possible to combine oligosaccharides with hydrogen gas generated by intestinal bacteria, and it is expected that the hydrogen power will be maximized.

[0282] In addition, sake lees fermentation products, re-fermented sake lees, sake lees decomposition products, sake lees processed products, or sake lees fermentation extract powder may be added. These sake lees-related materials are known to have the functions of reducing visceral fat, normalizing the intestinal environment, and absorbing oil and excreting lipids. In addition, they are known to have anti-stress effects, endurance-improving effects, and beauty effects. Therefore, by blending these with the reduced fermented lactic acid bacteria powder according to this embodiment, further synergistic effects are expected.

[0283] As described above, the reduced fermented lactic acid bacteria powder according to this embodiment contains lactic acid bacteria cells and lactic acid bacteria fermentation extract, which act to increase the number of good bacteria in the intestines, and can obtain bactericidal effects and antioxidant and anti-glycation effects against so-called "bad bacteria" in the body. This makes it possible to realize effective intestinal care and oral care, to realize anti-aging effects, and to be widely used for beauty purposes. Furthermore, by collaborating with ionized apatite, silk hydrogen pearl powder, soy isoflavone, and hydrocharcoal, it is possible to expect high synergistic effects in addition to the antioxidant and anti-glycation effects, such as effects on beautiful skin, beautiful hair, beautiful nails, and hair growth, and use in osteoporosis countermeasures and locomotor countermeasures by strengthening bones.

Claims

1. A composition containing a lactic acid bacteria production substance using a lactic acid bacteria fermentation extract comprising a lactic acid bacteria production substance, Contains brown sugar, soybean pulp, or soybean-derived lactic acid bacteria fermentation extract, Contains 22% or more by weight of plum extract, which is a plant of the Rosaceae family. Contains natural biological or mineral-derived hydroxyapatite and ionized apatite containing at least citric acid, Contains cherry blossom extract, The present invention comprises at least one of deep ocean water minerals, Great Salt Lake minerals, and lactic acid bacteria, and reduced fermented fulvic acid containing fulvic acid; A composition containing a lactic acid bacteria production substance, characterized by being fermented using hydrogen and deep sea water.

2. 2. The composition containing a lactic acid bacteria production substance according to claim 1, characterized in that the lactic acid bacteria fermentation extract contains 377 (mg / L) to 566 (mg / L) of peptides.

3. The lactic acid bacteria production substance-containing composition according to claim 1, further comprising at least one of water-soluble silicon, ceramide, hyaluronic acid, eggshell membrane peptide, nacre pearl powder, soybean isoflavones including equol or daidzein.

4. The composition containing a lactic acid bacteria production substance according to claim 1, further comprising 0.03 to 5% by weight of at least one or more of the following: extract of a plant of the Rosaceae family, hibiscus extract, cranberry extract, pomegranate extract, olive extract (leaves, fruit, flower), camellia extract (leaves, fruit, flower), perilla fruit extract, tea fruit extract or soybean extract.

5. 2. The composition containing a lactic acid bacteria production substance according to claim 1, further comprising silk hydrogen pearl powder obtained by combining silk powder and hydroxyapatite with hydrogen and deep sea water.

6. 2. The composition according to claim 1, further comprising curcumin and pea sprout extract.

7. 2. The composition according to claim 1, further comprising at least one of rose petal powder, grape seed powder, hibiscus powder, and cranberry extract.

8. 2. The composition according to claim 1, further comprising a barley lactic acid fermentation extract and a passion flower extract.

9. 2. The composition containing a lactic acid bacteria production substance according to claim 1, further comprising coconut shell activated carbon.

10. A combination of red orange extract, pineapple enzymes, papaya enzymes, hydrogen and deep sea water.

2. The composition according to claim 1, further comprising a combination of:

11. It is a combination of at least one of activated carbon or edible charcoal, inulin, hydrogen and deep sea water.

2. The composition containing a lactic acid bacteria production substance according to claim 1, further comprising combined hydrocharcoal.

12. The hydrocharcoal further contains kagome kelp powder and citric acid. The composition containing a lactic acid bacteria production substance according to claim 11, characterized in that

13. 2. The composition containing a lactic acid bacteria production substance according to claim 1, further comprising chlorogenic acid.

14. The present invention is characterized by further containing multiple types of oligosaccharides, lactic acid bacteria, and green papaya extract.

2. The composition according to claim 1, which contains a lactic acid bacteria production substance.

15. 2. The composition containing a lactic acid bacteria production substance according to claim 1, further comprising at least one of astaxanthin, an extract of Princess Matsutake, Coprino, Pleurotus ulmoides, and Antrodia camphorata.

16. The composition containing lactic acid bacteria production substances as described in claim 3, characterized in that the water-soluble silicon consists of at least one of plant-derived silicon, mineral-derived silicon, and water-soluble silicon derived from hot springs, spring water, lava, and rocks in the Fuji, Hakone, Kirishima, and Sakurajima areas.

17. A method for producing equol, comprising adding soybean isoflavones including daidzin or daidzein to the composition containing a lactic acid bacteria production substance according to claim 1 under specified conditions.

18. A method for producing a composition containing a lactic acid bacteria production substance using a lactic acid bacteria fermentation extract comprising a lactic acid bacteria production substance, comprising: Brown sugar, soybean pulp, or soybean-derived lactic acid bacteria fermentation extract, 22% or more by weight of plum extract from the Rosaceae family; A natural biological or mineral-derived hydroxyapatite and an ionized apatite containing at least citric acid; Cherry blossom extract, A reduced fermented fulvic acid containing at least one of deep ocean water minerals, Great Salt Lake minerals, and lactic acid bacteria, and fulvic acid; to form a mixture; and fermenting the mixture using hydrogen and deep sea water. A method for producing a composition containing a lactic acid bacteria production substance, comprising:

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