JP3945544B2 - Method for producing soybean extract - Google Patents

Description

【００４０】
殺菌状態
寒天に、肉エキスおよびペプトンを加え、直径９０ｍｍのシャーレに導入して細菌用培地を調製した。また、寒天に、ブトウ等およびペプトンを加え、直径９０ｍｍのシャーレに導入して真菌用培地を調製した。
【００４１】
実験サンプル１〜９から、０．１ｍｌの液を分取し、上記培地の表面に接種した。そして、細菌用培地は温度３７℃で４８時間、真菌用培地は温度２５℃で７日間培養した。
【００４２】
各々の培地を観察し、細菌或いは真菌が繁殖したかどうかを判定した。得られた結果を表１に示す。
【００４３】
【表１】

[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a soybean extract that does not inactivate the activity of a protein having an activity inhibiting activity of trypsin contained in a soluble component when a soluble component in soybean is extracted with an aqueous extraction solvent to produce a soybean extract. It relates to a manufacturing method.
[0002]
TECHNICAL BACKGROUND OF THE INVENTION
Infants or elderly people who use diapers have been in contact with the skin for a long time because of excrement, and people who have rashes, sores or itching around the anus and genital area, so-called diaper rash Not a few. Diaper rash is caused by direct irritation to the skin of stool itself or the propagation of various bacteria, and the cleaning of the affected area is effective for its treatment and prevention.
[0003]
Cleaning the affected area is cumbersome to clean with water, so various detergents including alcohol or surfactant are applied to the affected area as aerosols and oils, or soaked in a tissue etc. It is done by that. However, although these conventional detergents have excellent detergency, they are highly irritating and care must be taken so that they do not cause rashes themselves. Accordingly, there has been a demand for the development of a cleaning agent that can effectively remove the cause of diaper rash and that has little irritation to the affected area or the anus and genital areas.
[0004]
By the way, the fecal irritation that contributes to diaper rash is caused by intestinal proteolytic enzymes remaining in the feces, particularly proteases having trypsin-like proteolytic activity (hereinafter referred to as trypsin-like proteases). It has been found by recent research. Reducing the activity of this trypsin-like protease can be a very effective means for preventing or treating diaper rash.
[0005]
In recent years, it has been found that soybean contains a protein having an activity of inhibiting trypsin activity (hereinafter sometimes referred to as SbI) (Rosmarie Vogel et al., Natuerliche Proteinasen-Inhibitoren (1966), Georg). See Thieme Verlag, Stuttgart).
[0006]
Soybeans have long been processed into soybean extracts to make effective use of their edible value, and have been eaten as tofu or the like as they are or after being solidified. Specifically, such soybean extract is produced by adding soybean to water to swell, grinding, heating with a large amount of water, and then filtering. Moreover, when using the obtained soybean extract as a drink as it is, it is common to sterilize separately and to prevent decay.
[0007]
Such a soybean extract contains most of the protein in the soybean, minerals, vitamins, and the like, and is less irritating to the skin. Therefore, if it contains SbI and has the above-mentioned activity inhibiting action, diaper rash can be prevented. The present inventors considered that it would be effective as an active ingredient in quasi-drugs for treatment and prevention, such as detergents. However, it has been found that the soybean extract produced by the conventional method has a remarkably low trypsin activity inhibitory action and is not particularly effective from the viewpoint of removing the irritation of feces itself that causes diaper rash.
[0008]
The inventors of the present invention have solved the problems of the prior art and intensively studied and studied to produce a soybean extract having an activity of inhibiting trypsin. As a result, SbI was also extracted from the conventional soybean extract. However, SbI was denatured and inactivated by the harsh extraction conditions of the conventional method or the heat sterilization conditions after extraction, and thus the knowledge that the soybean extract has no trypsin activity inhibitory action was obtained, and the present invention was completed.
[0009]
OBJECT OF THE INVENTION
The present invention has been made in order to solve the problems associated with the prior art, and is a soybean that can be easily and mass-produced with a soybean extract that has little irritation to the skin and has an inhibitory effect on trypsin activity by SbI. It aims at providing the manufacturing method of an extract.
[0010]
SUMMARY OF THE INVENTION
In the method for producing a soybean extract according to the present invention, a soybean extract that absorbs water and grinds the swelled soybean and then heat-extracts in an aqueous solvent to produce the soybean extract.
The heat extraction treatment is performed in an aqueous extraction solvent having a pH of 1.0 to 12.0 at a temperature that does not inactivate the protein having a trypsin inhibitory action contained in the soybean extract. The heat sterilization treatment does not deactivate the protein having a trypsin inhibitory action contained in the soybean extract after adjusting the soybean extract to any one of pH 1.0 to 2.0 and pH 11.0 to 12.0, and It is performed at a temperature at which the soybean extract is substantially aseptic.
[0011]
In the present invention, the aqueous extractant, water may also be substantially neutral, which used as it is, for example, by adding at least one acidic compound selected from organic and inorganic acids in water, the It can be acidic in the pH range. The aqueous extraction solvent may be basic in the above pH range by adding at least one basic compound selected from basic organic and inorganic compounds to water .
[0012]
Specific examples of the acidic compound include inorganic acids such as hydrochloric acid and 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. Examples thereof include organic acids.
[0013]
Basic compounds include basic inorganic compounds such as sodium hydroxide, potassium hydroxide, calcium hydroxide and sodium dihydrogen phosphate, and basic organic compounds such as ammonia, triethanolamine, lysine, arginine and histidine. It can be illustrated.
[0014]
Moreover, when performing the said heat | fever extraction process, the said soybean extract is adjusted to acidic or basic suitably using the said acidic compound or basic compound. In this case, the heat extraction treatment may be performed in an aqueous extraction solvent having a pH of 3.0 to 12.0.
[0015]
In the present invention, pressurized heat sterilization process, the temperature 60 to 100 [° C., preferably at boiling temperature, it is desirable that made 0.1 to 60 minutes. Furthermore, in the method for producing a soybean extract according to the present invention, the heat extraction treatment is performed at a pH of 1. It may be performed at any of 0 to 2.0 or pH 11.0 to 12.0. In such a case, the heat sterilization treatment may not be performed. At this time, the heat extraction treatment is desirably performed at a temperature of 60 to 100 ° C., preferably at a boiling temperature.
[0016]
DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, the manufacturing method of the soybean extract which concerns on this invention is demonstrated more concretely.
In the method for producing a soybean extract according to the present invention, soybean which has absorbed water and swollen is heated and extracted in an aqueous extraction solvent adjusted to a specific pH value to obtain a soybean extract. It is adjusted to a specific pH value possible and sterilized by heating.
[0017]
Hereinafter, the heat extraction process and the heat sterilization process of the present invention will be described in more detail.
In the heat extraction process of the present invention, the soybean used as an extraction raw material is swollen by absorbing water, and is ground using a stone or pulverizer to add water to improve the extraction efficiency. It is desirable.
[0018]
The soybean raw material thus obtained is heated and extracted using an aqueous extraction solvent adjusted to pH 1.0 to pH 12.0, preferably pH 1.0 to 2.0 or pH 11.0 to 12.0.
[0019]
By heating the soybean raw material in such an aqueous extraction solvent in the pH range, other soluble components can be obtained without deactivating the protein having the activity of inhibiting trypsin contained in soybean (hereinafter referred to as SbI). At the same time, it can be extracted into an extraction solvent.
[0020]
In particular, when an aqueous extraction solvent having a pH of 1.0 to 2.0 or pH 11.0 to 12.0 is used, the obtained soybean extract is made substantially sterile by the bactericidal action of the solvent. Therefore, there is an advantage that the heat sterilization treatment is not necessary. Furthermore, when the solution in which SbI is dissolved has a pH of 1.0 to 2.0, SbI is particularly difficult to be denatured by heating, and thus can be heated at a higher temperature for a long time.
[0021]
The aqueous extraction solvent used in the present invention can be adjusted to 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. In addition, it is desirable that the acidic compound applied to the present invention hardly denatures SbI and does not have irritation to human skin. Therefore, from this point of view, it is desirable to refrain from using, for example, nitric acid, sulfuric acid, carboxylic acid, dehydroacetic acid, sorbic acid and propionic acid.
[0022]
Basic aqueous extractable solvents include basic inorganic compounds such as sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium dihydrogen phosphate and ammonia, and basic such as triethanolamine, lysine, arginine and histidine. It can be prepared by adding a basic compound such as an organic compound to water. Moreover, it is desirable that the basic compound applied to the present invention does not easily denature SbI and does not have irritation to human skin. Therefore, from such a viewpoint, for example, hypochlorite, chloroisocyanurate, chloramine T, bleaching powder, etc. are desirably refrained from use.
[0023]
These acidic compounds and basic compounds may be used alone or in combination, and the addition amount thereof is appropriately selected depending on the pH to be prepared.
[0024]
The amount of such an aqueous extraction solvent is not particularly limited as long as the raw material can be heated uniformly and a moderately viscous soy extract having an inhibitory effect on topsin activity can be obtained. It is used in an amount of 50 to 10,000 parts by weight, preferably 100 to 1000 parts by weight with respect to parts by weight.
[0025]
The heat extraction treatment of the present invention using the aqueous extraction solvent described above may be performed at a temperature and time at which SbI is not deactivated, specifically 60 to 100 ° C., preferably boiling temperature, usually 0.1 to 60. It is desirable to carry out for a minute, preferably 1 to 30 minutes.
[0026]
The soybean extract obtained by such a heat extraction treatment is subjected to the following heat sterilization treatment as it is as a mixture containing the soybean extract or after separation by filtration. As described above, when the heat extraction treatment is performed at either pH 1.0 to 2.0 or pH 11.0 to 12.0, the heat sterilization treatment may not be performed. The product can be made by adjusting the pH, for example, approximately neutral.
[0027]
The heat sterilization treatment in the present invention is performed under the condition that the soybean extract is sterilized and substantially aseptic without deactivating SbI. In such a heat sterilization process, the obtained soybean extract is performed at pH 1.0 to 2.0 or pH 11.0 to 12.0.
[0028]
The pH of the soybean extract can be adjusted by adding the acidic compound or basic compound used in the preparation of the aqueous extraction solvent.
Such heat sterilization is performed at 60 to 100 ° C., preferably at the boiling temperature, for 0.1 to 60 minutes, preferably 1 to 30 minutes.
[0029]
The soy extract obtained in this way is filtered as desired, and then adjusted to a suitable pH, for example, adjusted to about neutrality 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 substantially sterile can be produced easily and in large quantities.
[0030]
【The invention's effect】
As described above, according to the method for producing a soybean extract according to the present invention, a soybean extract having an activity of inhibiting trypsin activity can be produced easily and in large quantities. Since such a soy protein having an inhibitory effect on trypsin activity is less irritating to the skin, it is advantageous for quasi-drugs for preventing or treating skin diseases such as diaper rash caused by stimulation by the proteolytic activity of trypsin. Can be used.
[0031]
【Example】
Hereinafter, the manufacturing method of the soybean extract which concerns on this invention is demonstrated more concretely based on an Example.
[0032]
[Example 1]
Production of soybean extract Soybean was soaked in water at room temperature for 24 hours to absorb water, and ground using a pulverizer while adding water to prepare a soybean raw material.
[0033]
1000 g of this soybean raw material (dry weight: 365 g) was put into 2 liters of water as an extraction solvent and extracted 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.
[0034]
The obtained soybean extract was divided every 100 ml to prepare a plurality of samples, which were divided into 11 groups 1 to 11. Among these groups, groups 1 to 4 were adjusted to pH 1.0, pH 2.0, pH 3.0, and pH 6.0, respectively, by changing the amount of hydrochloric acid added.
[0035]
In addition, Group 5 of the sample was adjusted to pH 7.0 without adding anything, and Groups 6 to 11 were adjusted to pH 8.0, pH 10.0, pH 11.0, respectively, by changing the amount of sodium hydroxide added. The pH was set to 12.0, pH 13.0, and pH 13.7.
[0036]
Samples of each group 1 to 11 were heat sterilized at boiling temperature (100 ° C.) for 5 minutes.
Samples from each group 1-11 were adjusted to pH 7.8 as appropriate by addition of hydrochloric acid, sodium hydroxide or triethanolamine, and then assayed for the ability to inhibit proteolytic activity by trypsin and bactericidal status in the following manner .
Inhibition of proteolytic activity by soybean extract An enzyme solution containing trypsin was prepared in 0.001N hydrochloric acid at a concentration of 1 μg / ml.
[0037]
To 0.2 ml of this enzyme solution, 1.6 ml of triethanolamine buffer (pH 7.8) and 1 ml of benzoyl-L-arginine-P-nitroanilide (BAPA) aqueous solution (concentration 1 μg / ml), and samples 1 to 9 After adding 0.2 ml of the solution taken from either of them and incubating for 10 minutes at 37 ° C. with gentle shaking, 1 ml of 30% acetic acid aqueous solution was added to prepare a test solution.
[0038]
The absorbance of the obtained test solution at a wavelength of 405 nm was measured. The obtained results are shown in Table 1.
In addition, a control sample prepared by adding 1.6 ml of the above-mentioned triethanolamine buffer and 1 ml of an aqueous BAPA solution and 0.2 ml of distilled water to the enzyme solution was prepared and incubated in the same manner as in the above experimental samples 1 to 11, The absorbance at a wavelength of 405 nm was measured and found to be 1.758. The obtained absorbance value was regarded as a value of 100% trypsin enzyme activity, and the trypsin activity inhibition rate (%) of each sample was calculated by the following formula. The obtained results are shown in Table 1.
[0039]
[Expression 1]

[0040]
Sterilized state Meat extract and peptone were added to agar and introduced into a petri dish having a diameter of 90 mm to prepare a bacterial medium. Further, butto and the like and peptone were added to agar and introduced into a petri dish having a diameter of 90 mm to prepare a fungal medium.
[0041]
From the experimental samples 1 to 9, 0.1 ml of liquid was collected and inoculated on the surface of the medium. The bacterial medium was cultured at a temperature of 37 ° C. for 48 hours, and the fungal medium was cultured at a temperature of 25 ° C. for 7 days.
[0042]
Each medium was observed to determine whether bacteria or fungi had propagated. The obtained results are shown in Table 1.
[0043]
[Table 1]

Claims (4)

In the method of producing soybean extract by crushing swelled soybeans by absorbing water, followed by heat extraction treatment and heat sterilization treatment in an aqueous extraction solvent,
In the aqueous extraction solvent having a pH of 1.0 to 12.0, the heat extraction treatment does not deactivate the protein having a trypsin inhibitory action contained in the soybean extract, at a temperature of 60 to 100 ° C. for 0.1 to 60 minutes. Done within range,
After the heat sterilization treatment after the heat extraction treatment has adjusted the soybean extract to any one of pH 1.0 to 2.0 and pH 11.0 to 12.0, a protein having a trypsin inhibitory action contained in the soybean extract is added. A method for producing a soybean extract, characterized in that the method is carried out at a temperature of 60 to 100 ° C. for 0.1 to 60 minutes , wherein the soybean extract is not inactivated and substantially sterilized. After milling the swollen soybean which absorbs water, it has trypsin inhibitory action contained in soybean extract in any aqueous extraction solvent at pH 1.0-2.0 and pH 11.0-12.0 The manufacturing method of the soybean extract characterized by including the heat extraction process performed within the range of the temperature of 60-100 degreeC and 0.1-60 minutes which does not inactivate protein. The pH value of the aqueous extraction solvent or soybean extract is 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. The method for producing a soybean extract according to claim 1 or 2 , wherein at least one selected acidic compound is added to adjust the acidity . The aqueous extraction solvent or soybean extract has a pH value of at least one base 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 1 or 2 , wherein a basic compound is added to adjust the basicity .