KR101921736B1 - Extract method of sarcodon saparatus and use - Google Patents

Abstract

The present invention is based on the finding that the glycoprotein extracted by the mycelial growth method using ginseng as a medium to enhance mycelial growth of mycelium and mushroom mycelium and the ability to stimulate the immune system of excellent lectin, Through the activation of lipolysis ability, and completed the present invention
It is a main object of the present invention to provide an extract derived from a mushroom mycelium using ginseng as a medium.
In order to accomplish the above object, the present invention provides an extract obtained from a mushroom containing the lectin protein and saponin obtained by inoculating and cultivating a mycelium of mushroom in a ginseng culture medium.

Description

[0001] The present invention relates to a method for producing an extract from a mushroom,

More particularly, the present invention relates to an extract obtained from a mycelium of mushroom with ginseng as a medium, a method for producing the extract, and a use thereof.

The mushroom belongs to the family Mushroom, and it lives mainly in the broad-leaved forest. In addition, it has a distinctive flavor and is called mushroom, and it belongs to the taxonomic fungus. It is a kind of most bacillus species and contains a lot of nutrients such as carbohydrates, proteins, lipids, minerals and vitamins.

It has been used for various purposes due to the merits of the above-mentioned mushroom. For example, Japanese Patent Application Laid-Open No. 1997-0027302 discloses the use as a novel protease derived from a mushroom cultivated by culturing mycelium of mushroom mushroom and separating the mycelium from them.

In addition, Japanese Patent Registration No. 1074649 discloses a composition of a cosmetic composition for whitening and antioxidation comprising a mushroom extract and a maca extract, and suggests its use as a cosmetic product.

In addition, in the registered patent No. 859324, 9 kg of the mushroom was dried for 7 days in the shade to obtain 250 g of dried mushroom, and then 20 g of licorice, 200 g of bank, 50 g of persimmon leaf and 50 g of elm were put in a pressure cooker together with 25 L of water, And then boiled at 100 ° C. for 2 hours, then sterilized and vacuum-packed in a retort. The composition of the method of manufacturing a health drink containing mainly mushroom is disclosed. Suggesting its use.

That is, as described above, diaphor mushroom is utilized for various purposes such as protease, raw material for functional cosmetics, and functional beverage.

On the other hand, it is known that the mushroom exhibits various functional effects by the contained lectin. The lectin is a glycoprotein which is a protein that binds to a saccharide component, and is a term that refers to a substance produced by non- Which is mainly found in legumes and plants. Recently, however, new lectins have been discovered from mistletoe, mushroom and microorganisms.

Generally, lectin is called as aggregate or hemagglutinin because it binds specifically to the glycoprotein part of the surface of red blood cells. Therefore, it has been reported that other physiological activities such as leukocyte aggregation effect and leukocyte proliferation stimulation effect According to these characteristics, some lectins have been studied for their application as an activity system of Pysioloy immunotherapeutic agent.

In addition, immunotherapy has been actively studied by many researchers because it can expect maximum healing effect while minimizing damage to normal tissues. Specifically, cancer vaccine, monoclonal antibody, and cytokine microorganism products have.

On the other hand, Copolang has been developed and used from cloud mushroom fruiting body in Korea, and it belongs to a double microorganism product which is a protein polysaccharide of mushroom. In the case of Phellinus linteus mycelial culture extract, The system is being used as a product for activation.

The mushroom protein polysaccharide is a macromolecule whose molecular weight is usually 100,000 to several millions and is stable to heat and does not dissolve in an organic solvent. Since water can not be extracted at room temperature, it is common to use a heat extraction method in which the sample is heated under atmospheric pressure or high pressure for 1 to 10 hours and extracted.

However, it is expected that the mushroom is inhabited only in broad-leaved forests and has excellent protein and lipolytic properties, thus helping immune activity.

Accordingly, the present invention relates to a method for inhibiting the activation of the immune system of lectins, wherein the glycolipids extracted by the mycelial growth method using ginseng as a medium for obtaining the mycelium of mushroom, And activation of lipolysis ability, and completed the present invention

It is a main object of the present invention to provide an extract derived from a mushroom mycelium using ginseng as a medium.

It is also an object of the present invention to provide a method for producing an extract derived from a mushroom mycelium using ginseng as a medium.

It is also an object of the present invention to apply the extract extracted from the mycelium of mushroom with ginseng as a medium to the additive of the water base.

It is also an object of the present invention to apply a lectin protein extracted from a mycelium of Mycobacterium tuberosum as a medium for ginseng to an additive of a cosmetic lotion.

It is also an object of the present invention to apply an extract derived from the mycelia of Mycobacterium tuberus using ginseng as a medium to food additives.

In order to accomplish the above object, the present invention provides an extract obtained from a mushroom containing the lectin protein and saponin obtained by inoculating and cultivating a mycelium of mushroom in a ginseng culture medium.

The present invention also relates to a method for producing a microbial cell culture, which comprises: obtaining a culture extract obtained by adding an extraction solvent to a cultured mycelium and pulverizing it to obtain a culture extract; Obtaining a primary extract by concentrating the culture extract under reduced pressure to obtain a primary extract; And obtaining the final extract by dissolving the extract in a solvent and ultrafiltering to obtain a final extract.

Preferably, the method further comprises the step of obtaining a secondary extract in the same manner as in the step of obtaining the primary extract after the step of obtaining the primary extract.

More preferably, the extraction solvent is selected from the group consisting of water, 4 mM to 1 M Tris buffer at pH 7.0 to 9.0, 0.001 to 1.0 N NaOH solution, 0.001 to 1.0 N citric acid solution and 0.1 to 5.0% (w / v) ) ≪ / RTI > NaCl solution.

Further, the present invention provides the use of a water-borne additive containing the above extract as an active ingredient.

Further, the present invention provides the use of a cosmeceutical additive containing the above extract as an active ingredient.

Further, the present invention provides the use of an additive for food or beverage containing the above extract as an active ingredient.

According to the present invention, the extract obtained from the mushroom of the present invention can be used for the treatment and prevention of thrombosis by providing a high erythrocyte agglutination effect and an excellent solid fat decomposition effect by extracting the mycelium after proliferation of mycelium using the ginseng as a medium And can be used as an additive for cosmetic waters, as an additive for cosmetic lotion, and as an additive for food.

FIG. 1 is a flow chart of a method for producing an extract from a mushroom of the present invention,
2 is a photograph showing the growth of mycelium according to the present invention,
3 is a photograph of mycelium according to Example 3,
4 is a result according to the fourth embodiment,
5 is a compositional ratio of Example 5,
6 is a compositional ratio of Comparative Example 5,
7 is a result of Example 5,
8 is a result of Comparative Example 5,
9 is a compositional ratio of Example 6,
10 is a composition ratio of Comparative Example 6,
11 is a result of Example 6,
12 is a result of Comparative Example 6,
13 is a composition ratio of Example 7,
14 is a compositional ratio of Comparative Example 7,
Fig. 15 shows the results of Figs. 13 and 14. Fig.

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

As shown in FIG. 1, the method for producing the extract of the present invention comprises the steps of (S1) obtaining a culture extract (S1) obtained by adding a solvent to a cultured mycelium and pulverizing it to obtain a culture extract, (S2) of obtaining a first extract to obtain a first extract, a step (S2) of obtaining a second extract to obtain a second extract by concentrating the first extract at a reduced pressure, and a step (S4) of obtaining a final extract to obtain a final extract.

Each step will be described in detail below.

Step (S1) of obtaining a culture extract

The step of obtaining a culture extract is a step of culturing mycelia in a ginseng culture medium and then obtaining a culture extract.

First, 20 to 25 parts by weight of the saccharified maltose and 2 to 3 parts by weight of the salt of New Zealand are mixed with 100 parts by weight of fresh ginseng, and the mixture is stirred at 110 to 121 DEG C for 20 to 30 minutes for 1 to 20 minutes Sterilize by vacuum.

Then, the mycelium of Sacodon aspratus is inoculated and cultured at 25 to 35 ° C for 30 to 45 days to grow mycelium.

Then, it is subjected to secondary decompression sterilization at 110 to 121 캜 for 20 to 30 minutes in order to suppress the denaturation of the extract.

Extraction solvent is added to the autoclaved body and the artificial mycelium which have undergone reduced pressure sterilization and pulverized to obtain an extract.

Examples of the extraction solvent include water, 4 mM to 1 M Tris buffer solution at pH 7.0 to 9.0, 0.001 to 1.0 N NaOH solution, 0.001 to 1.0 N citric acid solution and 0.1 to 5.0% (w / v) NaCl solution May be used alone or in combination.

After culturing, the sterilized mycelia are immersed in the extraction solvent and stirred at room temperature (20 to 25 ° C) for 45 to 50 hours, followed by filtration or centrifugation to obtain a culture extract.

 Step (S2) of obtaining the first extract

The step of obtaining the first extract is a step of obtaining the first extract by using the additional procedure of the culture extracted extract.

 The extract obtained in the culture and extraction step is concentrated under reduced pressure and ammonium sulfate is added to obtain a first extract.

At this time, to maintain the lectin activity of saponin and mushroom which are useful components of ginseng, the concentration under reduced pressure is set at 50 to 60 ° C.

Ammonium sulfate is dissolved in the concentration of 20 to 100 g / L in the concentrated extract, and the mixture is allowed to stand at room temperature (20 to 25 ° C) for 20 to 28 hours, followed by centrifugation to obtain a primary extract as a supernatant.

 Step (S3) of obtaining the second extract

The step of obtaining the secondary extract is carried out in the same manner as the step of obtaining the primary extract, wherein the starting material proceeds to the primary extract.

The step of obtaining the secondary extract is a step for increasing the concentration of the extract and the yield of the whole step, and this step may be omitted if necessary.

Step (S4) of obtaining the final extract

The step of obtaining the final extract is a step of finally extracting the saponin-containing lectin from the second extract obtained through the step of obtaining the second extract.

100 to 200 parts by weight of ethanol as a primary extract is mixed with 100 parts by weight of the secondary extract obtained in the step of obtaining the secondary extract, and the mixture is allowed to stand at room temperature (20 to 25 ° C) for 20 to 28 hours , Ultrafiltration to obtain the final extract.

The final extract extracted in this manner consists of a lectin containing saponin free of impurities.

The following examples illustrate the present invention in more detail.

Example 1: Preparation of Lectins Derived from Mycelia

The well-washed ginseng was crushed to about 6 to 8 mm, and 22 parts by weight of the saccharified malachite and 2.5 parts by weight of the salt of New Zealand were mixed with 100 parts by weight of fresh ginseng and sterilized at 120 캜 for 30 minutes. Then, mycelial growth was carried out by inoculating cultured mycelium of mycelium of mushroom and culturing at 25 ~ 35 ℃ for 40 days.

The mycelium grown at this time is as shown in Fig.

Then, 1200 ml of Tris buffer solution (10 mM of pH 7.5) was added to 200 g of the above mycelium of hyacinths, and the mixture was allowed to stand at room temperature (20 to 25 ° C) for 24 hours and then filtered under reduced pressure to obtain a culture extract.

Thereafter, ammonium sulfate was dissolved at a concentration of 100 g / L, and the supernatant was obtained by centrifugation at 10,000 rpm at 10 ° C or less to obtain a first extract. The first extract was then diluted with ammonium sulfate at a concentration of 100 g / L And the mixture was allowed to stand at room temperature (20 to 25 ° C) for 24 hours. The mixture was centrifuged at 10,000 rpm at 10 ° C or lower to obtain a secondary extract through the supernatant. Ethanol was then added to 200 parts by weight After the addition, the mixture was allowed to stand at room temperature for 24 hours. Ultrafiltration was performed to obtain 590 g of final extract. The final extract was confirmed to contain 100 μg / ml of lectin and 2.5 μg / ml of saponin.

Example 2. Chemical analysis of lectins derived from mushroom

Total sugar content and total protein content contained in the final extract were measured. At this time, the total sugar content was analyzed according to the calibration curve by measuring the color development degree at 625 nm by reacting with Anthrong's reagent using glucose as a standard according to the Anthron method. The total protein content was determined by Coomassie brilliant blue) method was used to measure Bovine serum albumin as a standard protein at 620 nm.

As a result of the measurement, it was confirmed that the content ratio of sugar and protein was 1: 2 to 3.5 (w / w) because the total sugar content of lectin was 20 to 25% (w / w) and the total protein content was 40 to 70% (w / Respectively.

Example 3 Red blood cell coagulation effect of lectin derived from mushroom

 The coagulation effect of the erythrocytes obtained by the present invention was compared with that of the lectin (comparative example) of the mushroom, another mushroom

 In the same manner as in Example 1, the lectin component of the mycelia of mushroom was obtained at a concentration of 100 μg / ml, and the lectin aqueous solution obtained in Example 1 was diluted to 1, 2, 4, and 8-16 times, Peripheral blood of mice was diluted 10-fold with 5 IU / ml heparinized physiological saline and blood was prepared.

The blood dilution was added to 25 wells of 12 wells, 25 μg of each lectin diluent was added, and the coagulation effect of red blood cells was compared after incubation for 2 hours in a 35-37 ° C incubator.

As shown in FIG. 3, the concentration of the extract of the present invention was higher than that of the mushroom lectin.

 Example 4 Effect of Lipolysis of Lectin Derived from Mushroom

 For the measurement of the lipolysis ability of the extract obtained by the present invention, fat and lard were filtered to collect fat components. Uji and Donji were kept in a solid state at room temperature, and thus a liquid fat was produced at a temperature of 60 ° C or higher, and a comparison test was conducted with 100 μg / ml of the mushroom lactin component extracted from the example and 100 μg / ml of the mushroom lactin component.

 In order to compare the fat content and the degree of decomposition of the extract for the purpose of this comparative experiment, 90% by weight of each of Uji and lard and 10% by weight of each of the examples and comparative examples were added and mixed for 30 minutes, The tests were compared.

The results of the physical property test are shown in Fig. As shown in FIG. 4, the extract of the present invention has an ability to decompose solid fat.

Example 5 Effect as a cosmetic additive for water

Add the vegetable fatty acid, vegetable oil, glycerin, and purified water to 65 ℃ ~ 75 ℃ and add the potassium hydroxide. After the saponification process, the pH is adjusted with acidic citric acid. The extracts and fragrances obtained by the invention were added to prepare a cosmetic waterspout.

At this time, as shown in FIG. 5, a total of 1,500 kg of 300 Kg of each step was prepared at the mixing ratio of Example 5-1 to Example 5-5, and a comparative test was conducted.

As a comparative example, as shown in Fig. 6, five ordinary soaps containing chitosan were prepared.

The soaps of the prepared examples and comparative examples were used in the following manner to examine the skin condition.

13 points for skin condition, 4 points for recovery from bacterial skin, 3 points for hardness and 3 points for non-car. All points are 18 points for adult men and women between 18 and 25 years old The results were shown in FIG. 7 and FIG. 8, respectively. The results are shown in FIG. 7 and FIG. 8, respectively.

As shown in FIG. 7 and FIG. 8, it is confirmed that the effect of the embodiment is excellent. In other words, it was confirmed that the regenerating effect of the skin was exhibited within a short period of time as compared with the general beauty soap, and the recovery of the skin was accelerated additionally. However, the skin trouble occurred again in the period of transition After using the soap containing the additive according to the present invention, the effect of restoring the skin was obtained again.

It was confirmed that the inventive material had a high effect when the composition ratio of Example 5-2 or more, that is, 2% by weight or more, was contained.

Example 6 Effect as Lotion Additive

Vegetable oil, vegetable oil, glycerin, purified water, and the oil is dissolved well at 65 ° C to 75 ° C. After the emulsification process is completed by adding the emulsifier, the pH is adjusted with citric acid, which is an acidic agent, Base cosmetics lotion was prepared and adjusted to a compounding ratio in five steps. As shown in FIG. 9, 500 kg of each 100 kg of each step was prepared in Examples 6-1 to 6-2.

As a comparative example, chitosan-added lotion was prepared as shown in Fig.

Skin condition tests were carried out in the same manner as in Example 5, and as a result, the results of the skin lotion according to the present invention and those of the comparative example are shown in FIG. 11 and FIG. 12, respectively.

As a result of the test, it was confirmed that Examples 6-1 to 6-5 exhibited faster skin regeneration effects than Comparative Examples 6-1 to 6-5.

Example 7 Effect as Food Additive

The extract was prepared by adding the extract according to the present invention to Dongchimi of the food. At this time, 1 wt%, 2 wt%, 6 wt% and 10 wt% of the extract according to the present invention were added to the total weight to prepare Dongchimi (Examples 7-1 to 7-4) The composition ratio is shown in Fig. For comparison, as shown in Fig. 14, Comparative mushroom extracts were added at the same ratio to prepare Comparative Examples 7-1 to 7-4.

The protein degradation test is shown in Fig. As shown in FIG. 15, the proteolytic activity of the mushroom mycelium was more than ± 0.45% at the concentration of 2%, and that of the mushroom mycelium at the concentration of about 10% .

Further, taste evaluation and consumer taste were conducted by 20 evaluation teams, and it was confirmed that the degree of preference of the examples was higher.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, And all of the various forms of embodiments that can be practiced without departing from the technical spirit.

S1: Obtain culture extract S2: Obtain first extract
S3: Secondary extract obtained Step S4: Step of obtaining the final extract

Claims (7)

delete delete Obtaining a culture extract by inoculating and cultivating a mycelium of mushroom in a ginseng culture medium, adding an extraction solvent to the cultured mycelium and pulverizing it to obtain a culture extract;
Obtaining a primary extract by concentrating the culture extract under reduced pressure to obtain a primary extract; And
A step of obtaining a final extract by dissolving the extract in a solvent and ultrafiltering to obtain a final extract,
Further comprising a step of obtaining a second extract, which is carried out in the same manner as the step of obtaining the first extract, with the first extract obtained after the step of obtaining the first extract as a starting material,
Wherein the extraction solvent is selected from water, a 4 mM to 1 M Tris buffer solution at pH 7.0 to 9.0, a 0.001 to 1.0 N NaOH solution, a 0.001 to 1.0 N citric acid solution, and a 0.1 to 5.0% (w / v) Wherein the extract is at least one selected from the group consisting of:
delete An additive for a water-borne nutrient containing an extract prepared by the method according to claim 3 as an active ingredient.
An additive for cosmetics containing an extract prepared by the method according to claim 3 as an active ingredient.
An edible food additive comprising an extract prepared by the method according to claim 3 as an active ingredient.

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