JPS6130541B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention uses edible "natto" as a starting material, which is obtained by breeding natto bacteria on steamed soybeans and ripening them.Based on this, extracts are obtained and used in cosmetics and other foods. It is about application. Natto has been eaten in Japan since ancient times as one of the foods with high nutritional value.
However, the drawback was that long-term storage produced a strange odor, which meant that not all people liked eating it. Also, the stringy viscous substance peculiar to natto is something that many people avoid.
Therefore, natto itself has continued to be a food with a palatable aspect. Therefore, the present inventors removed the unpleasant odor in natto,
Initially, we thought of extracting the nutrients without damaging them and using them in food, and various studies were conducted. As a result, the extract, which is made mainly from the stringy viscous substances in natto, is mainly composed of protein, which is not only nutritionally superior, but also has an excellent moisturizing and lubricating effect, which is especially good for the skin. It has been found that since it exhibits a smooth lubricating effect when applied, it is advantageous not only for nutritional preservation but also for use in cosmetics. Therefore, the present inventors further investigated the application to cosmetics, continued research on the relationship between the extraction method, yield, and moisturizing effect, and completed the present invention as described below. Until now, there have been no other examples of using extracts from natto in cosmetics or for food, but the method of extracting from natto in the present invention is easy to operate. It is expected that new fields of use will be developed. In other words, even people who cannot eat natto itself can make drinks made from its extract or in granular form.
Furthermore, it can be easily used in applications other than cosmetics, such as by processing it by filling it into tablets or gelatin capsules, and mixing it with appropriate excipients and other nutritional supplements. [Example 1] Edible natto obtained by breeding natto bacteria on steamed soybeans and ripening them is placed in a moist heat type heat sterilizer in advance, and sterilized at a temperature of 60 to 70°C for about 1 hour. For 5 kg of processed natto, 5 to 15 kg of water
After adding and crushing it to form a kayu-like substance, the kayu-like substance is then filtered using a filter cloth or a cotton plug to obtain about 10 ml of a viscous filtrate. Therefore, next, for this filtrate, as a known solvent immiscible with water,
For example, esters such as chloroform, isoamyl acetate, isopropyl acetate, isobutyl acetate,
Alcohols such as n-hexyl alcohol and decyl alcohol, n-hexane, n-heptane,
Add about 10 to 50% of a type of solvent selected from hydrocarbons such as benzene, petroleum ether, and cyclohexane to the filtrate, stir well, and let stand in a cool dark place for about a day and night. Therefore, it is separated into an aqueous layer [layer A] and a solvent layer [layer B], and each is separated. The yield of layer A is approx.
10, the yield of B layer is about 5. Next, for the A layer portion, a water-miscible solvent is used. One type selected from known ethanol, methanol, acetone, propyl alcohol, etc. is added at a ratio of about 30 to 90% and stirred. After stirring, the mixture was left standing in a cool, dark place for about a day and night, and the separated precipitate layer [C layer] and supernatant liquid layer [D
layer] respectively. The fractionated yield is
The C layer weighs about 200-500 g, and the D layer weighs about 7.5-10 g.
The C layer is an extract mainly composed of highly viscous proteins, but the solvent and water used remain in this C layer, so by further removing this by vacuum distillation, It gradually becomes a rubbery, elastic solid. The yield of this solid is approximately
215-270 g is obtained. When this product is used in cosmetics or foods, it is best to disperse it in water again and use it as a suspension (emulsion). The amount used in this case does not need to be particularly limited, but it is 0.1 to 10% in terms of the solid substance mentioned above.
The degree is one guideline. [Example 2] When the total amount 5 of layer B separated (separated) in the process of Example 1 above is taken and the solvent is completely removed by distillation under reduced pressure, about 50 to 100 g of oils and fats remain. . This stuff has a peculiar smell (odor) that is typical of natto.
is transitioning. Then, in the same manner as in the refining process for edible liquid oil (soybean oil), saponification is performed with an alkali, and approximately 20 g of unsaponifiable matter is obtained. Therefore, in contrast, ether or n-hexane
After adding 500 to 1000 ml and thoroughly shaking, separate the solvent layer and perform vacuum distillation again to obtain the residue. This is an extract from which the comb has been completely removed and is mainly composed of phytosterols. The yield is about 7.5 to 18 g. [Example 3] Using the layer D separated during the process of Example 1 described above, the solvent is completely removed by distillation under reduced pressure to obtain about 30 to 50 g of solid matter. Therefore, next, for this solid matter, NaC, KC, Na ₂ SO ₄
Add 2.5 to 5 ml of one of these aqueous solutions (each with a concentration of 45 mo), collect the precipitate, and then add 50% weakly alkaline solution. After dissolving in a methanol solution, filtration is performed to collect the filtrate, and this filtrate is concentrated under reduced pressure to obtain about 2.5 to 9 g of an extract consisting of pigment components mainly composed of isoflavones. As mentioned above, the ratio of the total yield of each extract obtained in Examples 1 to 3 is 86 to 90% of the protein-based extract obtained from layer A and further through layer C. %, phytosterol-based extract obtained from layer B is 3-6%, from layer A,
Furthermore, the extract mainly composed of isoflavones obtained through the D layer is about 1 to 3%,
It was found that a total yield of about 250 to 300 g can be obtained from 5 kg of natto. The extracts obtained in the above examples can be used alone in cosmetics and foods, but
In particular, the protein-based extract obtained in Example 1 has a high viscosity and has a concentration of 50-50% in a concentrated liquid state.
Shows a value around 80cps. This viscosity indicates moisturizing and slipping properties, and in a mixture of ethanol and water, it mixes well with each of the extracts obtained in Examples 2 to 3, increasing the slipping properties. That is, it has a smooth, non-sticky feel, and has a moisturizing effect, and is easily used in cosmetics and foods. In addition, the taste is good, and in order to bring out the unique richness of natto, it is better to use extracts mainly composed of sterols obtained in Examples 2 and 3 or isoflavones mainly, rather than extracts mainly mainly composed of proteins. It is recommended to use an extract consisting of pigment components. Basically, it can be used as a simple lotion or drink by adding an appropriate fragrance or cooling agent and diluting it with water or ethanol, but the amount of extract added or content in this case is If the viscosity is adjusted to about 10 to 30 cps based on the viscosity value, the characteristic slippery filling exhibited by natto extract will be good. The moisturizing and lubricating effect is similar to that of acidic mucopolysaccharides such as hyaluronic acid, and it is characterized by no sticky feeling. In other words, unlike moisturizing effects based on animal-derived proteins such as collagen and albumin, it is non-sticky and provides a smooth feel. Next, in the above example, a method was adopted in which the natto raw material was heat sterilized in advance and then extracted. However, in order to obtain a highly viscous extract, the longer the time required for heat sterilization, the lower the I found out what to do. Therefore, in order to obtain a highly viscous extract, we decided to reexamine the sterilization effect. In other words, various studies have been conducted regarding sterilization operations, such as the intermediate process during extraction and the final process, but the workability in both cases is difficult due to bacterial growth, so it is necessary to sterilize the raw natto in advance. We found that the best method for extraction is based on this. Therefore, regarding the sterilization operation, we tried ethylene oxide gas sterilization, which does not affect the viscosity and is advantageous on an industrial production scale, but due to the elapsed time,
It was found that an extract with a rather high viscosity was obtained, and the yield was further increased. [Example 4] Using the same natto as used in the previous example, 100 g of each was placed in a high-speed film sterilization bag measuring 28 cm in length x 15 cm in width, and ethylene oxide gas was applied for a predetermined period of time as shown in Table 1. After sealing, a bacterial test (by agar plate pour method) was performed.
After determining the sterilization effect (Table 1), take out 100g of each ethylene oxide (EO) sterilized natto from the sterilization bag and add 1000g of purified water to each.
ml and stir at a temperature of 15±1°C.
When gently stirred at a rotation speed of 100 rpm for 1 hour, this resulted in the formation of natto on the surface.
A viscous substance mainly composed of stringy proteins is transferred into water. Therefore, next, separate this viscous aqueous solution, take the filtrate by suction filtration (using filter paper: Toyo Roshi No. 65), and add the same amount of ethyl ether to the yield of about 800 ml of filtrate. After the soaking, the aqueous layer to be separated is collected. When this aqueous layer is distilled under reduced pressure to remove water, about 5 to 10 g of solid material mainly composed of protein is obtained. Next, add acetone to about 100-200% of the solid material.
Finally, a dry powder could be obtained by thoroughly washing each sample using 1.0 mL of ml and distilling it under reduced pressure to remove acetone. The yields were as shown in Table 1. It was found that the yield increased and the viscosity increased with the passage of time during the ethylene oxide sterilization process.
Furthermore, the solubility in water also showed a tendency to be higher than that extracted from heat-treated natto. Table 1 shows the yield and viscosity of the protein-based extract extracted from natto by ethylene oxide sterilization. still,
The dissolution rate (%) was determined by accurately weighing 1 g of each protein-based extract (extract obtained in Example 4),
Pour into 250 ml of purified water at ℃, stir, and increase the rotation speed.
After 1 hour at 300 rpm to form an emulsified protein solution,
This liquid was forcibly filtered through a 0.8 micron membrane filter, and the resulting transparent solution was measured. In other words, the extract mainly composed of proteins extracted from natto obtained in Example 1 or Example 4 is well dispersed in water, but exhibits a milky white or milky yellow suspension. The liquid exhibiting a transparent aqueous solution, that is, the portion of the protein that is completely soluble in water, is very small.The dissolution rate shown in Table 1 is for this water-soluble protein, and the viscosity is also the same. It is caused by liquid. In other words, it was found that the unique viscosity of natto is mainly due to the protein part that is dispersed in water as an emulsion. On the other hand, as obtained in Example 4,
The viscosity of the dry powder of the protein-based extract itself, when dispersed in water, is approximately 100 cps, and the average viscosity is approximately 80-80 cps.
It shows a peak at 90cps. As a result, using natto that has been sterilized with ethylene oxide gas is not only more effective in sterilizing Bacillus subtilis than using heat-treated natto, but at the same time, the extension of the processing time also makes it easier to extract from natto. It was also found that the amount of viscous protein that was absorbed increased. At the same time, it was found that the dispersibility and solubility in water were also improved. The reason for this is that ethylene oxide is added to Bacillus subtilis, exhibiting a bactericidal effect, and also added to natto protein, which improves its solubility.
It is thought that this also showed an increase in yield.

【table】

[Reference prescription example]

(1) Beverage The protein-based extract obtained in Example 1 or 4 was dispersed in water and a small amount of ethanol,
Solution with viscosity adjusted to 50 cps... 1-30% Lactic acid... 0.2 Citric acid... 0.9 Sweetener... 3-10 Preservatives (parabens)... 0.1 Flavoring... Appropriate amount Add purified water. Set the total amount to 100. (2) Cosmetics <Lotion> A solution prepared by adding water to disperse the protein-based extract obtained in Example 1 or 4 and adjusting the viscosity to 30 cps... 5.0% spermaceti wax... 2.0 Beeswax... … 16.0 Liquid paraffin… 46.5 Cetyl alcohol… 2.0 Purified water… 26.8 Borax… 1.0 Fragrance and paraben (methyl)… Appropriate amount (3) <Vanishing cream> Stearic acid… 16.0% Sorbitan monostearate ...... 2.0 Polyoxyethylene sorbitan monostearate... 1.5 Protein-based extract obtained in Example 1 or 4... 3.5-4.5 Propylene glycol... 10.0 Fragrance and paraben (methyl)... 0.2 Purification Add water to make the total amount 100. (4) <Lotion> Ethanol... 9.0% Lactic acid... 0.2 Citric acid... 0.9 Sorbiturate... 4.0 Flavoring, coloring, preservative... Appropriate amount Mainly protein obtained in Example 1 or 4 An aqueous solution prepared by mixing 20% of the extract with 0.8% of the extract containing pigment components mainly composed of isoflavones obtained in Example 3... 20.0 Add purified water to bring the total amount to 100. (5) <Cold cream> Beeswax... 10.0% Ceresin... 10.0 Vaseline... 15.0 Lanolin... 5.0 Liquid paraffin... 17.5 Olive oil or rice germ oil... 10.0 Mainly the protein obtained in Example 1 or 4 Extract containing mainly phytosterols obtained in Example 2... 0.5 Extract containing mainly isoflavones obtained in Example 3... 0.2 Oryzanol... 1.0 Purified water... 22.7 Fragrance... 1.0 Preservative... 0.3 [Moisturizing effect] Next, looking at the moisturizing effect of the protein obtained in Example 1 or 4, it was dispersed in water in advance and the viscosity was adjusted to about 30 cps. using a solution,
This was further diluted 20 times and used as a test solution, and the amount of water that the test solution deviated from under the set relative temperature using a constant temperature and humidity machine was determined by the gravimetric method when a constant weight was reached. A 5% aqueous solution of sodium pyrrolidone carboxylate was used as a standard for comparison, and as shown in Figure 1, it was found that they had almost the same moisturizing effect. On the other hand, the following lotion was made using the 20-fold diluted solution used in the above test, and an application test was conducted on the skin of 40 women to check the additive-free lotion and the feeling of use. However, as shown in Table 2, the results showed that there was no sticky feeling, a good smooth feel, an excellent lubricity effect, and a refreshing feeling on use. (Formulation: lotion) Ethanol... 9.0% Lactic acid... 0.2 Citric acid... 0.9 Sorbiturate... 4.0 Diluted extract (viscosity 3-5 cps)... 8.0 Fragrance... 0.1 Dilute to 100% with purified water do.

〔safety〕

Regarding the safety of the extract according to the present invention, the starting material is natto, which is used for human consumption, and it is assumed that there are no problems.
To be sure, the protein-based extracts obtained in Examples 1 and 4 by oral administration to mice were dispersed in purified water.
The experiment was conducted using a solution whose viscosity was adjusted to around 30 sps, and the LD ₅₀ value was 40 ml or more, and it was evaluated to be highly safe without any problems. On the other hand, for the primary irritation test, a 48-hour patch test was conducted using the above-mentioned solution on the same 40 women as in the above-mentioned use sensitivity test.
No abnormalities such as erythema were observed. Furthermore, it is noteworthy that the protein-based extracts obtained in Examples 1 and 4 have a moisturizing and slipping effect as well as an inhibitory effect on tyrosinase activity; Interested in this, we conducted additional tests on each of the extracts obtained in Examples 1 to 4, and found that all of the extracts obtained in cases other than Example 2 had this effect. Ta. Therefore, the natto extract is advantageous because it can be expected to have a skin whitening effect as well as a moisturizing and lubricating effect. Table 3 shows Examples 1 to 4 of the present invention.
This figure shows the in vitro results of the melanin pigment production inhibiting effect of the natto extract obtained in . The reaction system in the test was L-tyrosine (1.0
mg/ml) 0.5ml, phosphate buffer (pH6.8) 2.0ml,
Distilled water or inhibitor solution (extract) 2.0ml, Cu ⁺⁺ ion (1% solution) 0.05ml, tyrosinase (1mg/
The reaction was allowed to proceed for 60 minutes in a constant temperature bath at 37.5°C using 1.0 ml of ml), and after completion, the absorbance at 640 nm was determined using a spectrophotometer to calculate the production rate. As a comparison sample,
Ascorbic acid (vitamin C) was used.

【table】

[Powder cosmetics]

(1) Dry powder obtained in Example 4... 1-70% Vitamin C... 0.3-30 Add starch to make a total amount of 100%. (2) Dry powder obtained in Example 4... 90-95% CMC or alginic acid... 5-10 (3) Extract obtained in Example 2... 1-2% 〃 3 〃... 0.3- 1 〃 4 〃... 80-95 Oryzanol (micronized product)... 0.2-0.3 Vitamin C... 1-3 (4) Dry powder obtained in Example 4... 95% Aloe polysaccharide powder (Veragel) -200)...
0.3 Vitamin C... 3-4 Oryzanol (micropowdered product)... 0.6-2.7 All of the powder type cosmetics mentioned above have high water absorption (hygroscopicity), so the filling container should be packed tightly. It is also easier to use if it is packaged in portions of about 0.1 to 2 g each. In addition, it can be added to foundations and packs, but in such cases, it is especially good to use it in combination with silk powder or low-molecular-weight peptides of silk. For cosmetics, for formulation with other cosmetic bases, the extracts obtained in Examples 1 to 4 are mixed in advance in any combination of water, water and ethanol, or water and ethanol. It is convenient to prepare a solution in which the compound is dispersed and dissolved in a solvent such as a polyol, and to mix the solution. On the other hand, in the extraction steps of Examples 1 to 4, natto that has been sterilized in advance by heating or ethylene oxide gas is used, and natto that has not been sterilized in advance is used for extraction. You can also do that. In that case, in the extraction process, the extraction process should be carried out at a temperature of around 20°C or lower, and in particular, for the obtained protein-based extract, concentrate it under reduced pressure and powder it. Alternatively, it has been found that it is desirable to sterilize the powder by sterilizing it with ethylene oxide gas in the final step after turning it into a powder by using a freeze dryer.

[Brief explanation of the drawing]

Figure 1 shows an aqueous solution of the protein-based extract from natto obtained in Example 1 or 4, with a viscosity adjusted to around 3 to 5 cps, but with no moisturizing effect. Graph showing. 1 is a diluted solution of the extract of the present application, and 2 is an aqueous solution containing 5% sodium pyrrolidone carboxylate.

Claims (1)

[Claims] 1. After heating or sterilizing natto with ethylene oxide gas, water is added to crush the natto, then filtered, and a water-immiscible solvent is added to the resulting filtrate and stirred, Leave to stand in a cool, dark place, take the separated aqueous layer after leaving to stand, then add a water-miscible solvent to the aqueous layer and stir, then leave to stand in a cool, dark place again, remove the precipitate, and under reduced pressure. After removing the solvent and water to obtain an extract mainly consisting of protein, or sterilizing the natto by heating or ethylene oxide gas,
Add water and crush, then filter, add a water-immiscible solvent to the obtained filtrate, stir, let stand in a cool dark place, take the separated aqueous layer after standing, and then A water-miscible solvent was added to the aqueous layer, stirred, and allowed to stand again in a cool, dark place.After standing, the separated supernatant layer was taken out, the solvent was completely removed under reduced pressure, and NaC, Add one of KC, Na ₂ SO ₄ to an aqueous solution, separate the precipitated components, and then add the precipitate to a weakly alkaline 50%
A method for producing a natto extract, which comprises dissolving it in methanol and then concentrating the obtained filtrate under reduced pressure to obtain a pigment component mainly composed of isoflavones. 2 After sterilizing natto with ethylene oxide gas,
Add water and stir gently to peel off the filamentous viscous material on the natto surface and transfer it to the water layer. Add the same amount of ethyl ether to the water layer from which the solids have been removed, and add enough ethyl ether. After shaking, the aqueous layer was separated, water was distilled off under reduced pressure, the obtained solid was thoroughly washed with acetone, and the acetone was distilled off again under reduced pressure. A method for producing natto extract, which is characterized by obtaining components mainly consisting of protein.