KR101192117B1 - Method for manufacturing of fermented ginseng or fermented red ginseng - Google Patents

Abstract

The present invention relates to a method for producing fermented ginseng or fermented red ginseng using aeration pellet culture method, in particular, a first step of preparing ginseng powder or red ginseng powder by grinding ginseng or red ginseng; A second step of sterilizing the ginseng suspension or red ginseng suspension by adding distilled water to the ginseng powder or red ginseng powder prepared in the first step; Preparation of 1 to 10 mm pellet mycelium by blowing 2 ~ 10vvm of air while shaking the culture of grass mushroom mycelium in Glucose-Yeast (GY) medium that maintains temperature of 30 ~ 35 ℃ and 100 ~ 400rpm Step 3; Inoculate the pellet mycelium prepared in the third step to the ginseng suspension or red ginseng suspension produced by sterilization in the second step, while maintaining the pH 4.5 ~ 6.5 at a temperature of 30 ~ 35 ℃ 2 ~ 10vvm A fourth step of preparing a fermented ginseng culture solution or a fermented red ginseng culture solution by injecting air into the culture; A fifth step of sterilizing the fermented ginseng culture solution or the fermented red ginseng culture solution prepared in step 4 by heating at 95 ° C. for 30 to 60 minutes; The sixth step of concentrating the fermented ginseng culture solution or fermented red ginseng culture solution sterilized in the fifth step; consisting of a large amount of ginsenoside metabolites such as Rh1, Rh 2, Rg3, Compound K having various physiological activities in a short period of time There is an effect that can be produced fermented ginseng or fermented red ginseng containing.

Description

Fermented ginseng or fermented red ginseng manufacturing method using aeration pellet culture method {Method for manufacturing of fermented ginseng or fermented red ginseng}

The present invention relates to a method for producing fermented ginseng or fermented red ginseng using aerated pellet culture method, and in particular, a method for preparing fermented ginseng or fermented red ginseng using aerated pellet culture method containing a large amount of ginsenoside metabolites having immunity enhancement and physiological activity within a short period of time. It is about.

When ginseng is ingested, non-polar alkaloids and the like are absorbed in the stomach, but most polysaccharides and saponins are not absorbed. They come in contact with strains that live in the gut, and these strains break down readily available sugar moieties and absorb them into the body. Ginsenoside metabolite, which is a form of sugar that is easily broken down and absorbed, is then absorbed into the blood to exert its effects on the human body. Most of these enterobacterial transformants or metabolites are significantly increased in efficacy compared to saponins prior to oral administration.

The main ingredient of ginseng is 'ginsenoside', a saponin, and ginsenosides Rb1, Rb2 and Rc, which are protopanaxadiols, and ginsenosides Re, Rg1, and Rf, which are protopanaxatriols. The pharmacological action of these ingredients is known, such as anti-cancer activity, anti-inflammatory, anti-diabetic action is known.

Therefore, various methods for cultivating ginseng have been proposed in order to contain a large amount of ginsenoside metabolites having various physiological activities as described above. It has a problem that less amount than cenoside expected.

The present invention has been made to solve the above problems of the prior art, using aeration pellet culture method that can contain a large amount of ginsenoside metabolites having a variety of physiological activities such as boosting immunity, antitumor, anticancer effect in a short period of time Its purpose is to provide fermented ginseng or fermented red ginseng manufacturing method.

Fermented ginseng or fermented red ginseng production method using a ventilated pellet culture method according to the present invention for solving the above problems is the first step of producing a ginseng powder or red ginseng powder by grinding ginseng or red ginseng; A second step of sterilizing the ginseng suspension or red ginseng suspension by adding distilled water to the ginseng powder or red ginseng powder prepared in the first step; Preparation of 1 to 10 mm pellet mycelium by blowing 2 ~ 10vvm of air while shaking the culture of grass mushroom mycelium in Glucose-Yeast (GY) medium that maintains temperature of 30 ~ 35 ℃ and 100 ~ 400rpm Step 3; Inoculate the pellet mycelium prepared in the third step to the ginseng suspension or red ginseng suspension produced by sterilization in the second step, while maintaining the pH 4.5 ~ 6.5 at a temperature of 30 ~ 35 ℃ 2 ~ 10vvm A fourth step of preparing a fermented ginseng culture solution or a fermented red ginseng culture solution by injecting air into the culture; A fifth step of sterilizing the fermented ginseng culture solution or the fermented red ginseng culture solution prepared in step 4 by heating at 95 ° C. for 30 to 60 minutes; And a sixth step of concentrating the fermented ginseng culture solution or the fermented red ginseng culture solution sterilized in the fifth step.

On the other hand, when the shaking culture in the third step by adding a mushroom fungus with grass fungus mycelium shake culture, the fungus mushroom, cloud mushroom, situation mushroom, Ganoderma lucidum mushroom, mushroom mushroom, shiitake mushroom, white mushroom, cordyceps, net At least one selected from mushrooms, mushrooms, chaga, thirsty mushrooms, willows, mandala and oyster mushrooms.

In the fourth step, air is injected from the lower side of the ginseng suspension or the red ginseng suspension toward the upper side through the air injection tube inserted to the lower side of the ginseng suspension or the red ginseng suspension.

Fermented ginseng or fermented red ginseng production method using the aeration pellet culture method of the present invention is configured as described above ginsenosides such as Rh1, Rh 2, Rg3, Compound K having a variety of physiological activities, such as immunity enhancement, antitumor, anticancer effect, etc. Contains a large amount of metabolites, there is an advantage that can significantly improve the body intake rate of ginsenosides.

In addition, there is an advantage that the culture days are shortened through aeration from the bottom of the ginseng suspension or red ginseng suspension.

1 is a flow chart showing a fermented ginseng or fermented red ginseng manufacturing method using aeration pellet culture method according to the present invention.
FIG. 2A is a ginsenoside analysis profile of the comparative examples listed in Table 2, and FIG. 2B is a ginsenoside analysis profile of the examples described in Table 2. FIG.
Figure 3 A is a diagram showing the solid cancer size of the control group shown in Table 3, B is a diagram showing the solid cancer size of the sample administration group described in Table 3.

Hereinafter, with reference to the accompanying drawings an embodiment of the fermented ginseng or fermented red ginseng production method using aeration pellet culture method according to the present invention will be described in detail.

1 is a flow chart showing a fermented ginseng or fermented red ginseng manufacturing method using aeration pellet culture method according to the present invention.

2A is a ginsenoside analysis profile of the comparative example shown in Table 2, and FIG. 2B is a ginsenoside analysis profile of the example shown in Table 2. FIG.

In addition, A of Figure 3 is a diagram showing the solid cancer size of the control group shown in Table 3, B is a diagram showing the solid cancer size of the sample administration group described in Table 3.

Fermented ginseng or fermented red ginseng manufacturing method using aeration pellet culture method according to the present invention is completed through a few steps.

First, the first step (S10) is a powdering process for preparing ginseng powder or red ginseng powder by grinding ginseng or red ginseng. Taking red ginseng powder as an example, the first step (S10) is to prepare red ginseng containing 15% or less moisture by washing, steaming and drying the ginseng, and then putting the red ginseng into a grinder.

The second step (S20) is a process of sterilizing the ginseng suspension or red ginseng suspension by adding distilled water to the ginseng powder or red ginseng powder prepared in the first step (S10). At this time, the added distilled water is an amount corresponding to 2 to 10 times (preferably 4 to 6 times) the weight of the ginseng powder or red ginseng powder. Then, the sterilization operation is performed by putting ginseng suspension or red ginseng suspension for about 15 to 60 minutes in a sterilizer maintaining 120 to 125 ° C.

The third step (S30) is 2 ~ 10vvm (preferably 3 3) while shaking the mycelium mycelium shaken for 2-5 days in Glucose-Yeast (GY) medium that maintains the temperature of 30 ~ 35 ℃ and rotation speed of 100 ~ 400rpm It is a process of making pellet mycelium of 1 ~ 10mm by blowing air of ~ 5vvm). At this time, GY (Glucose-Yeast) medium may be inoculated with only the mushroom fungus mycelium and aspirated while shaking to culture pellet pellets, but inoculated with GY (Glucose-Yeast) medium with the mushroom fungus mycelium in a shake culture Pellet mycelium can also be manufactured by doing this. The fungi mentioned here are at least one of cloud mushrooms, situation mushrooms, ganoderma lucidum mushrooms, matsutake mushrooms, shiitake mushrooms, white mushrooms, cordyceps, net mushrooms, mushroom mushrooms, chaga mushrooms, tree mushrooms, willow mushrooms, mandala mushrooms and oyster mushrooms. It refers to the selected mushroom mycelium.

More specifically, the third step (S30) is a GY (Glucose-Yeast) medium inoculated with grass mushroom mycelium (or biliary fungus) in a Erlenmeyer flask, while maintaining the Erlenmeyer flask at 30 ~ 35 ℃ 100 By rotating at ~ 400rpm and aeration of 2 ~ 10vvm is carried out, at this time, the aeration is directed to the upper side from the lower side of the triangular flask. That is, the outside air is first supplied to the lowest side of the Erlenmeyer flask by the air injection tube inserted and extended to the lower side of the Erlenmeyer flask, and the supplied air flows from the lower side of the Erlenmeyer flask toward the upper side. In this specification, this is referred to as 'breath core culture'.

The fourth step (S40) is inoculated with the pellet mycelium prepared in the third step (S30) to the ginseng suspension or red ginseng suspension produced by sterilization in the second step (S20), pH 4.5 at a temperature of 30 ~ 35 ℃ It is a process of preparing fermented ginseng culture or fermented red ginseng culture by blowing 2 ~ 10vvm (preferably 3 ~ 5vvm) air for 2-7 days while maintaining ~ 6.5.

At this time, the pellet mycelium inoculated in the ginseng suspension or red ginseng suspension is 5 ~ 20% of the weight of the ginseng suspension or red ginseng suspension.

When the air is injected into the ginseng suspension or the red ginseng suspension, that is, when the air is aerated, air is injected from the lower side of the ginseng suspension or the red ginseng suspension toward the upper side through an air injection tube inserted to the lower side of the ginseng suspension or the red ginseng suspension. It is to carry out 'breath culture' as mentioned above. For such a deep core culture is used a fermentable core fermentation such as bubble incubator or air lift incubator. The reason for such a deep core culture is that the aeration speed is faster than any other conventional methods.

The fifth step (S50) is a process of sterilizing the fermented ginseng culture solution or fermented red ginseng culture solution prepared in the fourth step (S40) by heating at 95 ° C for 30 to 60 minutes.

The sixth step (S60) is a process of filtering the fermented ginseng culture solution or fermented red ginseng culture solution sterilized in the fifth step (S50) and concentrating using a concentrator.

From now on, the present invention provides a more detailed embodiment of the method of preparing fermented ginseng or fermented red ginseng using the aeration pellet culture method according to the present invention as well as the embodiment of the present invention so that the characteristics of the embodiments of the present invention can be more easily understood. The comparative example which can be compared is shown.

<Examples>

Wash ginseng, steam and dry to prepare red ginseng with less than 15% water content.

The red ginseng is then pulverized through a grinder and powdered. 100 g of powdered red ginseng powder is added to 400 g of distilled water to make a red ginseng suspension, adjusted to pH 6.2, and sterilized at 121 degrees for 15 minutes.

Using a 500 mL triangular flask equipped with aeration core culture device for the grass mushroom mycelium, 5mm pellet mycelium is prepared while blowing 3vvm aeration while shaking culture for 3 days at 33 ° C and 300rpm in GY (glucose-yeast) medium.

The inoculated sterilized grass mushroom mycelium pre-cultivated in the form of 5mm pellets at 10% of the total weight of the red ginseng suspension was sterilized under pressure sterilization at 3 ° C and initial pH 6.2 at 33 ° C for 7 days.

The fermented red ginseng culture prepared in the above process was heat sterilized at 95 ° C. for 30 minutes, and 500 mL of 80% ethanol was added to the fermented red ginseng culture to reflux for 3 hours to extract ginsenoside, and the reflux was concentrated to 50 mL.

Comparative Example

The red ginseng suspension in <Example> was carried out in the same manner as in <Example>, except that the green mushroom mycelium was not treated.

<Test Example 1> pH, total sugar content, acidic sugar (uronic acid) content of fermented red ginseng culture solution using red ginseng powder

The pH, total sugar and acidic sugar content of the fermented red ginseng culture solution of the Example and the red ginseng suspension of the comparative example were measured by the following method, and the results are shown in Table 1 below. First, pH was measured by pH automatic measuring device, total sugar was measured by phenolsulfuric acid method, acidic sugar (uronic acid) content of Blumenkrantz and AsboeHansen method (Blumenkrantz, N.and AsoeHansen, G. (1973) Anal.Biochem , 54, 484).

DAY Example Comparative example pH Total sugar content
(mg / mL) Uronic acid
(ug / mL) pH Total sugar content
(mg / mL) Uronic acid
(ug / mL) 0 6.2 453 225 6.2 453 225 One 6.2 431 223 6.2 453 225 3 5.4 367 240 6.2 453 225 5 5.1 219 276 6.2 453 225 7 5.1 135 322 6.2 453 225

As shown in Table 1, the pH was reduced from 6.2 to 5.1, and the total sugar content was reduced from 453 mg / mL to 135 mg / mL in the case of <Example>, which was fermented / cultivated for 7 days using the grass mushroom mycelium. Uronic acid increased from 225 ug / mL to 322 ug / mL. Here, the decrease in the total sugar content as the fermentation proceeds is a result showing that the sugar of red ginseng suspension was used as a carbon source necessary for the growth of mushroom mycelium involved in fermentation. In addition, since the acid sugar contained in red ginseng is known as an immunoactive substance such as anti-cancer, it is preferable that the acidic sugar conjugate uronic acid increases as the fermentation progresses.

In contrast, <Comparative Example> did not process the grass mushroom mycelium, so fermentation / cultivation did not proceed at all, so there was no change in pH, total sugar content and uronic acid content even after one week.

Test Example 2 Measurement of Total Ginsenoside Content and Metabolite Content of Fermented Red Ginseng Culture Solution Using Red Ginseng Powder

Using ginseng powder, the total ginsenoside content and metabolite content of the fermented red ginseng culture medium of the Examples and Comparative Examples were measured. For ginsenoside analysis of the fermented red ginseng culture solution of the regular examples and comparative examples were analyzed by the following method using HPLC, and the results are shown in Table 2.

To explain the analysis in more detail, after weighing 10 g of the fermented red ginseng culture solution of the Example and the red ginseng suspension of the comparative example, respectively, 50 mL of 80% ethanol was added and extracted twice in a shaker at 80 ° C., using Whatman # 1 paper. Filtered. The filtrate was dried under reduced pressure using a vacuum condenser and dissolved by adding 50 ml of distilled water.

Then, 2 ml of diethyl ether was added and mixed with 1 ml of distilled water, and the mixture was centrifuged at 1500 rpm for 10 minutes to remove diethyl ether. Then, 1.5 ml of unsaturated butanol was added thereto, followed by mixing. Was recovered. 1.5 mL of distilled water was added to the recovered butanol layer, which was collected by repetition three times, and the mixture was centrifuged to remove the water layer.

The butanol layer thus obtained was dried at 40 ° C., 0.5 ml of methanol was added thereto, and then filtered through a 0.45 μm membrane filter to use as a sample for HPLC analysis.

In HPLC analysis, the column was used for Waters Breeze system HPLC equipped with C18 (250 X 4.6 mm) and the mobile phase was acetonitrile (Sol. A, HPLC grade, Sigma) and distilled water (Sol. B, HPLC grade) was used and the detector was an ELSD detector. The column temperature was measured by setting the flow rate at 1.0 mL / min and the UV wavelength at 203 nm at room temperature.

Gin Senocide Comparative Example (mg / g) Example (mg / g) Rb1 4.52 2.08 Rb2 2.32 1.63 Rc 2.52 1.71 Rk 0.35 3.78 Rh2 0.26 0.35 Rg1 3.53 0.12 (S) Rg3 1.52 3.53 (R) Rg3 1.13 2.71 Rg5 0.73 5.26 Compound k 0 1.09

<Test Example 3> Immune effect of fermented red ginseng broth using red ginseng powder

Compound K, Rg3, Rh1, and Rh2, which are known as ginsenoside metabolites in fermented red ginseng broth using red ginseng powder, are known to prevent cancer and inhibit the growth of cancer cells. The purpose of this study was to investigate the immune effects of red ginseng suspension fermented with grass mushroom strains. .

Experimental animals received ICR mice from Biolink and adapted them in animal laboratories while feeding and feeding water freely. The experimental animals had a temperature of 23 ㅁ 1 ℃ and a humidity of 55 ㅁ 5%. The feed was pellet-type experimental animal feed. The cancer cell line was inoculated with Sarcoma-180 cells in the abdominal cavity of 8-12-week-old ICR mice with a 1 ml syringe, and 1 ml of yellow ascites was collected. The stock solution was then inoculated into the normal ICR mice by 0.1 ml. It was raised for 28 days. Sample administration was divided into a control group and a sample administration group. The control group was orally administered a fixed amount of distilled water daily for 2 weeks, and the sample-administered group was completely dissolved in fermented red ginseng culture solution at 200 mg / kg / day in distilled water and orally administered at the same daily concentration. . The anticancer effect of Sarcoma-180 solid cancer was measured 28 days after the solid cancer of the mouse was extracted and the weight was measured by microbalance. Inhibition rate for solid cancer proliferation was calculated as follows, and the results are shown in Table 3.

% Inhibition = (1-Cancer cell weight in control group / cancer cell weight in control group) X 100

Sample Dose (mg / kg) Turmor weight Inhibition rate (%) Control group  - 11.3 0 Sample administration group  200 3.4 70

S10: First step S20: Second step
S30: third step S40: fourth step
S50: fifth step S60: sixth step

Claims (7)

A first step (S10) of preparing ginseng powder or red ginseng powder by grinding ginseng or red ginseng;
A second step (S20) of sterilizing after making ginseng suspension or red ginseng suspension by adding distilled water to the ginseng powder or red ginseng powder prepared in the first step (S10);
Preparation of 1 to 10 mm pellet mycelium by blowing 2 ~ 10vvm of air while shaking the culture of grass mushroom mycelium in Glucose-Yeast (GY) medium that maintains temperature of 30 ~ 35 ℃ and 100 ~ 400rpm Step 3 (S30) and;
Inoculate the pellet mycelium prepared in the third step (S30) to the ginseng suspension or red ginseng suspension produced by sterilization in the second step (S20), while maintaining the pH 4.5 ~ 6.5 at a temperature of 30 ~ 35 ℃ A fourth step (S40) of preparing a fermented ginseng culture solution or fermented red ginseng culture solution by incubating the air for 2 to 10 vvm for 7 days;
A fifth step (S50) of sterilizing the fermented ginseng culture solution or the fermented red ginseng culture solution prepared in the fourth step (S40) by heating at 95 ° C. for 30 to 60 minutes;
Fermented ginseng or fermented red ginseng using the aeration pellet culture method, characterized in that consisting of; sixth step (S60) to concentrate the fermented ginseng culture solution or fermented red ginseng culture solution sterilized in the fifth step (S50).
The method according to claim 1,
Fermented ginseng or fermented red ginseng production method using aeration pellet culture method characterized in that the shaking culture in addition to the fungi mushroom mycelium and shaker fungi when the shaking in the third step (S30).
The method according to claim 2,
The fungus is one or more mushrooms selected from cloud mushrooms, situation mushrooms, ganoderma lucidum mushrooms, matsutake mushrooms, shiitake mushrooms, white mushrooms, Cordyceps sinensis, net mushrooms, mushroom mushrooms, chaga mushrooms, tree mushrooms, willow mushrooms, mandala mushrooms and oyster mushrooms Fermented ginseng or fermented red ginseng production method using aerated pellet culture method characterized in that the mycelium.
The method according to claim 1,
The distilled water added in the second step (S20) is the weight of the ginseng powder or red ginseng powder fermented ginseng or fermented red ginseng production method using aeration pellet culture method, characterized in that 2 to 10 times the weight of the powder.
The method according to claim 1,
The pellet mycelium inoculated in ginseng suspension or red ginseng suspension in the fourth step (S40) is 5 to 20% by weight of the ginseng suspension or red ginseng suspension, fermented ginseng or fermentation using aerated pellet culture method Red ginseng manufacturing method.
The method according to claim 1,
In the third step (S30) while putting a GY (Glucose-Yeast) medium into the Erlenmeyer flask to shake culture, while blowing the outside air,
External air is fermented ginseng or fermented red ginseng manufacturing method using aeration pellet culture method characterized in that the flow from the lower side to the upper side of the triangular flask by the air injection tube inserted into the lower side of the Erlenmeyer flask.
The method according to claim 1,
In the fourth step (S40) through the air injection tube inserted to the lower side of the ginseng suspension or red ginseng suspension fermented ginseng using aeration pellet culture method characterized in that the air is injected from the lower side to the upper side of the ginseng suspension or red ginseng suspension Fermented red ginseng manufacturing method.

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