JPH09176033A - Production of soybean extract - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for producing a soybean extract, capable of easily and massively producing the soybean extract little in irritation to skins and having a trypsin activity-inhibiting action due to SbI. SOLUTION: This method for producing the soybean extract comprises grinding soybeans swollen by the absorption of water and subsequently subjecting the ground product to a heating extraction treatment using an aqueous medium. Therein, the heating extraction treatment is carried out in an aqueous extraction solvent having a pH of 1.0-12.0 at a temperature not inactivating a trypsin- inhibiting action-having protein contained in the soybean extract. If desired, a thermal sterilizing treatment after the heating extraction treatment is carried out at a temperature inactivating the trypsin-inhibiting action-having protein contained in the soybean extract and substantially sterilizing the soybean extract, after the pH of the soybean extract is controlled to either of 1.0-2.0 and 11.0-12.0.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to the production of a soybean extract by extracting a soluble component in soybean into an aqueous extraction solvent, so that the activity of a protein having a trypsin activity-inhibiting activity contained in the soluble component is detected. The present invention relates to a method for producing soybean extract that is not inactivated.

[0002]

BACKGROUND OF THE INVENTION Infants and elderly people who use diapers have long-term excrement contact with the skin, which causes symptoms such as rash, soreness and itchiness around the anus and genital area, so-called diapers. Many people have a rash. Diaper rash is caused by direct irritation of feces itself to the skin, propagation of various bacteria, etc., and cleaning of the affected area is effective for treatment and prevention.

Since cleaning with water is troublesome for cleaning the affected area, various cleaning agents including alcohol or surfactant are applied to the affected area in the form of aerosol or oil or soaked in a tissue or the like. It is done by wiping it off. However, although these conventional detergents are excellent in detergency, they have strong irritation, and care must be taken not to cause irritation by themselves. Therefore, there has been a demand for the development of a cleaning agent capable of effectively removing the cause of diaper rash and less irritating to the affected area or the affected area of the anus and genital area.

[0004] By the way, the stimulation of feces themselves, which is one of the causes of diaper rash, is caused by intestinal proteolytic enzymes in the feces, particularly proteases having a trypsin-like proteolytic activity (hereinafter referred to as trypsin-like protease). Recent research has revealed that there is. Reducing the activity of this trypsin-like protease can be a very effective means for preventing or treating diaper rash.

In recent years, it has been found that soybean contains a protein having a trypsin activity inhibitory activity (hereinafter sometimes referred to as SbI) (Rosmarie Vogel).
et al., Natuerliche Proteinasen-Inhibitoren (196
6), Georg Thieme Verlag, Stuttgart).

Soybeans have long been processed into soybean extracts in order to effectively utilize their edible value, and the soybeans have been eaten as tofu or the like as they are or after being solidified. Specifically, such a soybean extract is produced by soaking soybeans in water to swell, grinding, heating with a large amount of water, and then filtering. When the obtained soybean extract is used as a beverage as it is, it is generally sterilized separately to prevent spoilage.

Most of the proteins in soybeans, minerals, vitamins, etc. are extracted from such soybean extract, and there is little irritation to the skin.
The present inventors have considered that if it contains bI and has the above-mentioned activity inhibitory action, it will be effective as an active ingredient of a quasi drug for treating and preventing diaper rash, for example, a detergent.
However, it was found that the soybean extract produced by the conventional method has a remarkably low activity-inhibiting effect of trypsin, and is not particularly effective from the viewpoint of removing the irritation of feces themselves which causes diaper rash.

The inventors of the present invention have made extensive studies and studies to solve the problems of the prior art and produce a soybean extract having a trypsin activity-inhibiting effect, and as a result, SbI was not found in the conventional soybean extract. Although it has been extracted, due to the severe extraction conditions of the conventional method or the heat sterilization conditions after extraction,
The present invention has been completed based on the finding that soybean extract has no trypsin activity inhibitory action because SbI is denatured and inactivated.

[0009]

SUMMARY OF THE INVENTION The present invention has been made in order to solve the problems associated with the prior art as described above, and it is easy to obtain a soybean extract which has less irritation to the skin and has an activity of inhibiting the activity of trypsin by SbI. It is an object of the present invention to provide a method for producing soybean extract that can be produced in large quantities.

[0010]

SUMMARY OF THE INVENTION The method for producing soybean extract according to the present invention comprises:
After grinding the swollen soybean by absorbing water, in a method for producing a soybean extract by heat extraction treatment in an aqueous solvent,
The heat extraction treatment is carried out in an aqueous extraction solvent having a pH of 1.0 to 12.0 at a temperature at which a protein having a trypsin inhibitory action contained in soybean extract is not inactivated, and, if desired, after the heat extraction treatment. Heat sterilization
Soy extract pH 1.0-2.0 and pH 11.0-
After being adjusted to any of 12.0, it is characterized in that it is carried out at a temperature at which the protein having a trypsin inhibitory action contained in the soybean extract is not inactivated and the soybean extract is substantially sterilized.

In the present invention, the aqueous extraction solvent may be substantially neutral using water as it is. For example, water is water, and at least one acidic compound selected from organic acids and inorganic acids. Can be added to make it acidic in the above pH range. In addition, the aqueous extraction solvent is at least one selected from water-to-water basic organic and inorganic compounds.
It is also possible to add a basic compound of a kind to make it basic in the above pH range.

As the acidic compound, specifically, hydrochloric acid,
Inorganic acids such as boric acid, citric acid, acetic acid, butyric acid, tartaric acid,
Examples thereof include malic acid, phosphoric acid, phthalic acid, acetic anhydride, aspartic acid, asparagine, glutamic acid, glutamine, and benzoic acid.

As the basic compound, basic inorganic compounds such as sodium hydroxide, potassium hydroxide, calcium hydroxide and sodium dihydrogen phosphate, ammonia,
Basic organic compounds such as triethanolamine, lysine, arginine and histidine can be exemplified.

When the heat extraction treatment is carried out, the soybean extract is adjusted to be acidic or basic by using the acidic compound or basic compound as appropriate. At this time, the heat extraction treatment is performed at pH 3.0 to pH 12.
It may be carried out in 0 aqueous extraction solvent.

In the present invention, the heat sterilization treatment is carried out at a temperature of 60 to 100 ° C., preferably at a boiling temperature of 0.1 to 6
It is desirable to be performed for 0 minutes. Further, in the method for producing a soybean extract according to the present invention, the heat extraction treatment has a pH of 1.
5 to 2.0 or pH 11.0 to 12.0 may be performed, and in such a case, heat sterilization may not be performed. At this time, the heat extraction treatment is preferably performed at a temperature of 60 to 100 ° C., preferably a boiling temperature.

[0016]

DETAILED DESCRIPTION OF THE INVENTION The method for producing soybean extract according to the present invention will be described in more detail below. In the method for producing a soybean extract according to the present invention, the soybean that is swollen by absorbing water,
The soybean extract is obtained by heat extraction treatment in an aqueous extraction solvent adjusted to a specific pH value, and if desired, this is adjusted to a specific pH value capable of sterilization and subjected to heat sterilization treatment.

The heat extraction treatment and heat sterilization treatment of the present invention will be described in more detail below. In the heat extraction treatment of the present invention, soybean used as an extraction raw material,
It absorbs water and swells, and in order to improve its extraction efficiency, it is desirable to grind while adding water using a stone drier or a crusher.

The soybean raw material thus obtained has a pH of
1.0 to pH 12.0, preferably pH 1.0 to 2.0
Alternatively, it is extracted by heating using an aqueous extraction solvent adjusted to pH 11.0 to 12.0.

In an aqueous extraction solvent having such a pH range,
By heating the soybean raw material, the protein (hereinafter referred to as SbI) contained in soybean and having the activity to inhibit the activity of trypsin (hereinafter referred to as SbI) can be extracted into the extraction solvent together with other soluble components without being inactivated.

In particular, when an aqueous extraction solvent having a pH of 1.0 to 2.0 or a pH of 11.0 to 12.0 is used, the soybean extract obtained is substantially obtained by the bactericidal action of the solvent. Since it can be sterilized, it has an advantage that heat sterilization does not have to be performed. Furthermore, SbI is
When the solution in which it dissolves has a pH of 1.0 to 2.0, it becomes particularly difficult to denature by heating, and thus it becomes possible to heat at a higher temperature for a long time.

The aqueous extraction solvent used in the present invention can be adjusted to be acidic or basic in the above range by adding an appropriate acid or base to water. For example, acidic aqueous extraction solvents include inorganic acids such as hydrochloric acid and boric acid, and citric acid, acetic acid, butyric acid, tartaric acid, malic acid, phosphoric acid, phthalic acid, acetic anhydride, aspartic acid, asparagine, glutamic acid, glutamine, benzoic acid. It can be prepared by adding an acidic compound such as an organic acid to water. It is desirable that the acidic compound applied to the present invention is less likely to denature SbI and has no irritation to human skin. Therefore, from this point of view, for example, nitric acid, sulfuric acid, carboxylic acid,
Dehydroacetic acid, sorbic acid and propionic acid etc.
It is advisable to refrain from using it.

The basic aqueous extractable solvent includes basic inorganic compounds such as sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium dihydrogen phosphate, and ammonia, triethanolamine, lysine, arginine, histidine, etc. It can be prepared by adding a basic compound such as the basic organic compound of 1. to water. Further, it is desirable that the basic compound applied to the present invention is less likely to denature SbI and has no irritation to human skin. Therefore, from this point of view, it is desirable to refrain from using, for example, hypochlorite, isocyanuric chloride, chloramine T, bleaching powder and the like.

These acidic compounds and basic compounds are
Each may be used alone or in combination, and the addition amount thereof is appropriately selected and used depending on the pH to be adjusted.

The amount of such an aqueous extraction solvent used is not particularly limited as long as the raw material can be uniformly heated and a soybean extract having a moderately viscous effect of inhibiting topsin activity can be obtained, but it is usually ground. 50 to 10,000 parts by weight, preferably 100 to 10 parts, relative to 100 parts by weight of soybean raw material
Used in an amount of 00 parts by weight.

The heat extraction treatment of the present invention using the above-mentioned aqueous extraction solvent may be carried out at a temperature and for a time at which SbI is not inactivated, specifically at 60 to 100 ° C., preferably at boiling temperature, usually at 0. 1 to 60 minutes, preferably 1 to
30 minutes is desirable.

The soybean extract obtained by such heat extraction treatment is subjected to the next heat sterilization treatment as it is as a mixture containing it or after being separated by filtration.
In addition, as described above, the heat extraction treatment is performed at pH 1.0 to
When it is carried out at 2.0 or pH 11.0 to 12.0, the heat sterilization treatment may not be carried out, and after filtration, the pH is appropriately adjusted, for example, adjusted to be approximately neutral to obtain a product. be able to.

The heat sterilization treatment in the present invention is carried out under the condition that the soybean extract is sterilized without deactivating SbI and becomes substantially sterile. In such heat sterilization treatment, the obtained soybean extract is treated with a pH of 1.0 to 2.0 or a pH of 11.
It is performed as 0-12.0.

The pH of the soybean extract can be adjusted by adding the above-mentioned acidic compound or basic compound used for the preparation of the aqueous extraction solvent. Such heat sterilization treatment,
60-100 ° C, preferably at boiling temperature, 0.1-60
Minutes, preferably 1 to 30 minutes.

The soybean extract thus obtained is filtered, if desired, and then the pH is appropriately adjusted, for example, to be substantially neutral to obtain a product. According to the method for producing a soybean extract according to the present invention described above, a soybean extract having a trypsin activity inhibitory action by SbI and being substantially sterile can be easily produced in a large amount.

[0030]

As described above, according to the method for producing a soybean extract of the present invention, a soybean extract having a trypsin activity inhibiting action can be easily produced in a large amount. Since soybean protein having such trypsin activity inhibitory action has less irritation to the skin, it is advantageous for quasi-drugs for preventing or treating skin diseases caused by irritation by the proteolytic activity of trypsin, such as diaper rash. Can be used.

[0031]

EXAMPLES Hereinafter, the method for producing soybean extract according to the present invention will be described more specifically based on examples.

[0032]

Example 1 Production of Soybean Extract Soybean was immersed in water for 24 hours at room temperature to absorb water, and ground with a pulverizer while adding water to prepare a soybean raw material.

1000 g of this soybean raw material (dry weight: 36
5 g) was added to 2 liters of water as an extraction solvent,
Extraction was performed at a temperature of 100 ° C. for 10 minutes while stirring. The mixture obtained by this heat extraction treatment was filtered to obtain about 2 liters of crude soybean extract.

The soybean extract thus obtained was divided into every 100 ml to prepare a plurality of samples, which were divided into 11 groups 1 to 11. Group 1 to 4 of these, pH 1.0, pH 2.0, pH 3.0, respectively, by changing the amount of hydrochloric acid added
And pH 6.0.

Group 5 of the samples was adjusted to pH 7.0 without adding anything, and Groups 6 to 11 were adjusted to pH 8.0 and p by changing the amount of sodium hydroxide added.
H10.0, pH11.0, pH12.0, pH13.
PH 0 and pH 13.7.

The samples of each group 1 to 11 were heat-sterilized at the boiling temperature (100 ° C.) for 5 minutes. Samples from each group 1-11 were adjusted to pH 7. by appropriately adding hydrochloric acid, sodium hydroxide or triethanolamine.
After adjusting to 8, the ability to inhibit proteolytic activity by trypsin and the bactericidal state were assayed by the following method. Inhibition of Proteolytic Activity by Soybean Extract An enzyme solution containing trypsin at a concentration of 1 μg / ml in 0.001N hydrochloric acid was prepared.

To 0.2 ml of this enzyme solution, 1.6 ml of triethanolamine buffer (pH 7.8), 1 ml of benzoyl-L-arginine-P-nitroanilide (BAPA) aqueous solution (concentration 1 μg / ml), and sample 1 0.2 ml of the liquid collected from any one of ~ 9 was added, and the mixture was incubated at 37 ° C for 10 minutes while gently shaking, and then 1 ml of 30% acetic acid aqueous solution was added to prepare a test solution.

The absorbance of the obtained test solution at a wavelength of 405 nm was measured. Table 1 shows the obtained results. Also,
To the enzyme solution, add the above-mentioned triethanolamine buffer solution 1.6.
ml and a BAPA aqueous solution (1 ml) and distilled water (0.2 ml) were added to prepare control samples, and the above experimental samples 1 to 1 were prepared.
Incubation was carried out in the same manner as in 1, and the absorbance at a wavelength of 405 nm was measured and found to be 1.758. The obtained absorbance value is taken as the value of 100% trypsin enzyme activity,
The trypsin activity inhibition rate (%) of each sample was calculated by the following formula. Table 1 shows the obtained results.

[0039]

[Equation 1]

Sterilized state Meat extract and peptone were added to agar, and the diameter was 90 mm.
Was introduced into a petri dish to prepare a bacterial medium. In addition, butto and the like and peptone were added to agar and the mixture was introduced into a petri dish having a diameter of 90 mm to prepare a fungal medium.

From experimental samples 1 to 9, 0.1 ml of the liquid was collected and inoculated on the surface of the above medium. And the bacterial medium is at 37 ° C for 48 hours, and the fungal medium is at 25 ° C.
For 7 days.

Each medium was observed to determine whether bacteria or fungi grew. Table 1 shows the obtained results.

[0043]

[Table 1]

Claims (11)

[Claims] 1. After grinding the swollen soybean by absorbing water,
In the method for producing a soybean extract by heat extraction treatment in an aqueous solvent, the heat extraction treatment is carried out in an aqueous extraction solvent having a pH of 1.0 to 12.0 to extract a protein having a trypsin inhibitory action contained in the soybean extract. The soybean extract is subjected to a heat treatment for a time that does not deactivate it, and if necessary, a heat sterilization treatment after the heat extraction treatment is carried out so that the soybean extract has pH of 1.0 to 2.0 and pH of 11.
After being adjusted to any of 0 to 12.0, it is carried out by heating for a time during which the protein having a trypsin inhibitory activity contained in the soybean extract is not inactivated and the soybean extract is substantially sterilized. A method for producing soybean extract. 2. The soybean extract according to claim 1, wherein the aqueous extraction solvent is adjusted to be acidic by adding at least one acidic compound selected from organic acids and inorganic acids to water. Manufacturing method. 3. The aqueous extraction solvent is adjusted to be basic by adding at least one basic compound selected from basic organic and inorganic compounds to water. Method for producing soybean extract. 4. The soybean extract is adjusted in pH value by adding an acidic compound.
The method for producing a soybean extract according to. 5. The pH value of the soybean extract is adjusted by adding a basic compound.
3. The method for producing a soybean extract according to item 3. 6. The heat extraction treatment comprises pH 3.0 to pH 1.
The method for producing a soybean extract according to claim 1, wherein the method is carried out in an aqueous extraction solvent of 2.0 and the heat sterilization treatment is carried out. 7. The method for producing a soybean extract according to claim 4, wherein the soybean extract is heat-sterilized at a temperature of 60 to 100 ° C. for 0.1 to 60 minutes. 8. The heating and extraction treatment has a pH of 1.0 to 2.0.
8. The method for producing a soybean extract according to claim 7, which is carried out using an aqueous extraction solvent having a pH of 11.0 to 12.0. 9. The heat extraction treatment is performed at a temperature of 60 to 100 ° C.
The method for producing a soybean extract according to claim 8, wherein 10. The acidic compound is selected from hydrochloric acid, boric acid, citric acid, acetic acid, butyric acid, tartaric acid, malic acid, phosphoric acid, phthalic acid, acetic anhydride, aspartic acid, asparagine, glutamic acid, glutamine and benzoic acid. It is at least 1 type, The manufacturing method of the soybean extract of Claim 2 or 4 characterized by the above-mentioned. 11. The basic compound is at least one selected from sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium dihydrogen phosphate, ammonia, triethanolamine, lysine, arginine and histidine. The method for producing a soybean extract according to claim 3 or 5, which is characterized.