KR101823585B1 - Methods for preparing fermented ginseng extracts - Google Patents

Abstract

More particularly, the present invention relates to a method for producing a fermented ginseng extract, which comprises fermenting a ginseng extract with lactic acid bacteria, subjecting the fermented ginseng extract to high-pressure emulsification, and then inoculating and culturing a strain of Trichoderma reesei to obtain a fermented ginseng extract And a method for producing the fermented ginseng extract.
According to the present invention, a fermented ginseng extract containing a high content of ginsenosides Rd, Rg3-R and Rk1 can be prepared by the above production method. Through the production process of the present invention, specific ginsenosides Rd, Rg3- R and Rk1 saponins can be produced with increased fermented ginseng concentrate and ginseng product.

Description

Methods for preparing fermented ginseng extracts

The invention then relates to a method of manufacturing the fermented ginseng extract, and more specifically to lactic acid bacteria fermentation of ginseng extract high-pressure emulsification treatment Trichoderma riseyi (Trichoderma The present invention also relates to a method for producing a fermented ginseng extract comprising the step of obtaining a fermented ginseng extract by inoculating and culturing the fermented ginseng extract.

Ginseng has been known about 11 species, widely used, primarily for health promotion for thousands of years past, including the country in China and Japan, using the earth as a perennial sukgeuncho belonging to the genus Panax ginseng plants and trees classified haksang Acanthopanax (Aralaiaceae). Recently, it has been known that red ginseng, which contains a small amount of ginsenoside useful for the human body, is superior in the efficacy of steam ginseng steamed several times. Saponin, a representative active ingredient of ginseng, is a kind of compound called ginsennoside. Saponin, which is the main effect of ginseng extract extracted with water and ethanol, consists of more than 30 kinds of ginsenosides. Ginsenoside is a major component of ginseng extract and has been reported to have immunity, anti-inflammatory, anti-allergic, anti-cancer effect, hypotensive action, anti-cholesterol action, antithrombotic action, have.

Saponins ingested in the body are degraded by microorganisms in the intestine and absorbed into the body [Panta Med. 62, pp 453-457, 1996]. In the case of ginsenoside Rh1, it is sequentially converted to ginsenoside Rh, compound K, and protopanaxadiol by intestinal microorganisms. Intestinal microorganisms decomposing saponin of ginseng differ in the presence or absence and presence of the microorganism depending on the constitution and eating habits of the ginseng, and there is a difference in conversion to metabolites, so that there may be a difference in the efficacy after taking ginseng . In addition, since intestinal microflora vary from person to person, the conversion rate and bioavailability of ginsenoside are different [J. Pharm. Pharmacol., 50, pp1155-1160, 1998]. Therefore, in order to eliminate the difference between the efficacy and the absorption rate due to differences in intestinal microorganisms, the use of lactic acid bacteria fermented product of ginseng extract using microorganisms has an effect of overcoming individual differences and increasing the body absorption rate.

As a result, the inventors of the present invention have found that when ginseng extract fermented using lactic acid bacteria is treated with a high-pressure emulsifier and re-cultured as a Trichoderma reesei strain and heat-treated, It is possible to produce a fermented ginseng extract having an increased ginsenoside content of a specific ingredient, thereby completing the present invention.

Accordingly, it is a main object of the present invention to provide a method for preparing a fermented ginseng extract having increased contents of ginsenosides Rd, Rg3-r and Rk1.

Another object of the present invention is to provide a fermented ginseng extract having increased contents of ginsenosides Rd, Rg3-r and Rk1 produced by the above method.

According to one aspect of the present invention, the present invention provides a method for preparing a fermented ginseng extract comprising the steps of:

a) preparing ginseng extract from ginseng;

b) fermenting the ginseng extract with lactic acid bacteria;

c) destroying the lactic acid bacteria cells by subjecting the fermented lactic acid bacteria to high-pressure emulsification; And

d) Inoculating and culturing Trichoderma reesei in the high-pressure emulsified lactic acid bacteria fermented product to obtain a fermented ginseng extract.

The present inventors have sought to develop a method for converting saponin contained in ginseng, that is, ginsenoside, into a more useful form in the human body. As a result, the ginseng extract was fermented by inoculation with lactic acid bacteria, and after high pressure emulsification treatment, it was cultivated as Trichoderma reesei strain, and it was confirmed that the ginsenoside unique to the ginseng was converted into a specific form of ginsenoside useful for the human body.

The present invention relates to a method for increasing the content of ginsenoside-specific ginsenosides, namely ginsenosides Rd, Rg3-r and Rk1.

In the present invention, the term saponin has the same meaning as ginsenoside, i.e. ginsenoside, unless otherwise specified.

In the present invention, 'ginseng' is used to mean all ginseng processed in various forms such as ginseng, minced, white and red ginseng, and the method of the present invention can be applied to these various types of ginseng. It is apparent to those skilled in the art that these various types of ginseng may differ in the ingredient content of ginsenoside, but the effect of increasing the specific ingredient by going through the manufacturing steps of the present invention is the same.

The origin of ginseng used in the present invention is not particularly limited, and for example, domestic, American, Japanese, Himalayan, Vietnamese and Chinese acids can be used.

The ginseng extract used in the present invention can be easily prepared from ginseng using a known plant extraction method. For example, hot water extraction, organic solvent extraction and the like can be used. Specifically, the dried ginseng product or the pulverized product thereof is put into an extraction solvent and immersed at 5-40 DEG C for 1-15 days, It can be extracted by heating at 100 ° C for 4-20 hours. The solvent used for the extraction may be water, an organic solvent or a mixture thereof. The organic solvent may be ethanol, methanol, propylene glycol, butylene glycol, glycerin, hydrous ethanol, hydrolyzed butyleneglycol, , Ethyl acetate and the like can be used. The amount of the solvent used for the extraction is usually 3 to 20 times (weight ratio) of the raw material for extraction. In addition, the ginseng extract used in the present invention may further be a ginseng extract that has been subjected to a usual cooling, filtration, concentration, and sterilization processes.

In the method for producing the fermented ginseng extract of the present invention, the lactic acid bacteria include various lactic acid bacteria known in the art. For example, the lactic acid bacteria are Lactobacillus bacteria (Lactobacillus), Lactococcus (Lactococcus), flow Pocono stock (Leuconostoc), Pro are pleased tumefaciens (Propionibacterium), Enterococcus (Enterococcus), Bifidobacterium (Bifidobacterium), Streptococcus ( Streptococcus , and Pediococcus . Preferably, the lactic acid bacterium used in the present invention is Lactobacillus fermentum .

In the method for preparing the fermented ginseng extract of the present invention, the high-pressure emulsification treatment in step c) is performed 2 to 4 times at 1300 to 1600 bar. In the present invention, the lactic acid bacterial cells are disrupted by the high-pressure emulsification treatment to increase the content of crude fat, crude protein, carbohydrate and water-soluble dietary fiber in the fermented lactic acid bacteria (see Table 1). According to the change of nutritional composition of ginseng extract, inoculation of Trichoderma reesei in fermented lactic acid bacteria subjected to high pressure emulsification and cultivation shows that ginsenoside of specific ingredient is increased.

In the method for producing the fermented ginseng extract of the present invention, the cultivation of Trichoderma reesei in step d) can be carried out under suitable conditions. As in the following examples, the Trichoderma viridis strain can be inoculated into fermented lactic acid bacteria subjected to high-pressure emulsification and cultured at 25 to 30 DEG C for 3 to 5 days.

The content of ginsenoside Rd, Rg3-r and Rk1 in the fermented ginseng extract prepared by the method of the present invention was significantly lower than that of the ginsenosides of the existing ginseng extract, Respectively. In addition, when the fermented ginseng extract prepared by the method of the present invention further comprises a step of heat-treating at 110 to 125 ° C for 20 to 40 minutes after the step d), the decrease in the content of ginsenoside Rb1 and the decrease in the content of Rd, Rg3- r and Rk1 (see Table 4).

In the method for producing a fermented ginseng extract of the present invention, the content of ginsenosides Rd, Rg3-r and Rk1 in the fermented ginseng extract of step d) is increased by at least two times as compared with the ginseng extract of step a) do. The content of Rb1 was decreased from 10.51 mg / g to 2.23 mg / g about 4.7 times as compared with that of the first ginseng extract of the fermented ginseng extract prepared by the method of the present invention, while the content of Rd was 2.53 The content of Rg3-r was increased from 0.92 mg / g to 2.13 mg / g approximately 2.3 times, and the content of Rk1 was increased from 1.14 mg / g to 3.02 mg / g 2.6 times (see Table 4).

In particular, when the fermented ginseng extract according to the present invention is further subjected to the heat treatment step d), the content of Rb1 is 10.51 mg / g to 1.53 mg / g as compared with the ginsenoside content of the first ginseng extract The content of Rg3-r increased from 0.92 mg / g to 3.42 mg / g approximately 3.7-fold, while the content of Rk1 increased from 3.23 mg / g to 8.23 mg / g, (Rd), Rg3-r and Rk1 contents were increased by about 3.6-fold increase from 1.14 mg / g to 4.19 mg / g (see Table 4).

According to another aspect of the present invention, there is provided a fermented ginseng extract having increased contents of ginsenosides Rd, Rg3-r and Rk1 produced by the above method.

The content of ginsenosides Rd, Rg3-r and Rk1 was greatly increased in the fermented ginseng extract of the present invention. In the case of the present invention, the contents of ginsenosides Rd, Rg3-r and Rk1 are increased up to three times or more. The increase of the content of ginsenosides Rd, Rg3-r and Rk1 is attributed to the change of nutrients by the lactic acid bacteria fermentation and high pressure emulsification treatment of the first ginseng extract, followed by inoculation and cultivation of Trichoderma reesei . The microbial conversion of the ginsenoside component through this series of processes was found for the first time in the present invention.

As described above, according to the present invention, when the ginseng extract is cultured with lactic acid bacteria, treated with a high-pressure emulsifier, and then cultivated with Trichoderma reesei, fermented ginseng extract having an increased content of ginsenosides Rd, Rg3-R and Rk1 is prepared . Through the manufacturing method of the present invention, it was possible to remarkably improve the transplantability of Trichoderma reesei in the ginseng extract and to produce the fermented ginseng concentrate and the ginseng product in which the content of ginsenosides Rd, Rg3-R and Rk1 was increased.

FIG. 1 is a schematic view illustrating a method for producing a fermented ginseng extract according to the present invention.
FIG. 2 is a photomicrograph of the ginseng extract before and after the culture of lactic acid bacteria after high-pressure emulsification treatment in the manufacturing method according to the present invention.
FIG. 3 is a graph comparing the content of ginsenosides in the fermented ginseng extract according to the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for illustrating the present invention, and thus the scope of the present invention is not construed as being limited by these embodiments.

Comparative Example : Manufacture of ginseng extract

30 kg of 6-year-old ginseng was heat-treated at 90 to 100 ° C for 3 to 5 hours, and then 8.5 kg of the heat-treated ginseng was dried in a hot air dryer at 50 to 60 ° C for 3 days. Four to five times as much ionized water was added to the recovered ginseng extracts and extracted three times at 80 to 90 ° C for 8 hours. The extracted liquid was concentrated to obtain 5.0 kg of a ginseng extract having a solid content of 60%.

Production Example: Preparation of Fermented Ginseng Extract

30 kg of 6-year-old ginseng was heat-treated at 90 to 100 ° C for 3 to 5 hours, and then 8.5 kg of the heat-treated ginseng was dried in a hot air dryer at 50 to 60 ° C for 3 days. Four to five times as much ionized water was added to the recovered ginseng extracts and extracted three times at 80 to 90 ° C for 8 hours. The extracted liquid was concentrated to obtain a ginseng extract having a solid content of 60%. 5.0 kg of ginseng extract with a solid content of 60% was added to the fermentation tank and 15 L of triple ion water was added. The solution was sterilized at 100 ° C for 30 minutes and then cooled to 37 ° C.

200 mL (1.5 x 10 ⁸ cfu / ml) of Lactobacillus fermentum KCCM109129, which had been cultured for 2 days at 37 ° C in a MRS (deMan Rogosa Sharpe; Becton, Dickinson and Company) medium, After inoculation, the cells were fermented for 2 days under the condition of 0.5 bar, 37 캜, 100 rpm fermentation tank. The fermentation broth was sterilized at 100 DEG C for 30 minutes and cooled to prepare lactic acid-fermented ginseng extract.

The resulting lactic acid bacteria-extracted ginseng extract was passed through a high-pressure emulsifier (Microfludizer (110Y), Microfluidics Correlation) under the condition of 1,500 bar and three times, and then subjected to centrifugation at 28 ° C and 160 rpm in a potato dextrose broth (Becton, Dickinson and Company) (5.4 x 10 < ⁸ > cfu / ml) of the preculture of Trichoderma reesei BLJH304 cultured for 2 days was inoculated into the 10 L lactic acid fermented ginseng extract and cultured at 28 rpm for 3 days at 200 rpm and 0.5 vvm Respectively. The culture was heat-treated at 110 to 125 ° C. for 30 minutes, and 400 g of a filter aid was added thereto, followed by filtration with a filter press. The filtrate was concentrated to a solid content of 60 to 65% to prepare a fermented ginseng extract. The above process is schematically shown in Fig.

Experimental Example  One: High pressure oil painting  Confirmation of nutritional change by treatment

In order to analyze the changes in nutritional composition of the ginseng extract and the fermented ginseng extract prepared in the above Comparative Examples and Preparation Examples by high pressure emulsification treatment, the samples of each preparation step were lyophilized, and the crude fat, crude protein , Carbohydrates, sodium. Sugars, and soluble dietary fiber contents were analyzed. In order to measure the nutrient content of the sample, the Soxhloet extraction method for crude fat, the Macro-Kjeldalhl method for crude protein, the Schoorl method for reducing sugar, the Mohr method for sodium, the Molish reaction method for sugar, and the AOAC method for water- Respectively.

Changes of Nutritional Components by High Pressure Emulsification Treatment
Manufacturing stage Crude fat
(g / 100 g) Crude protein
(g / 100 g) carbohydrate
(g / 100 g) salt
(g / 100 g) sugars
(g / 100 g) receptivity
Dietary Fiber
(%)  1. Ginseng Extract 0.2 9.5 53.2 0.12 27.7 7.1  2. Cultured ginseng extract cultured with lactic acid bacteria 0.0 7.9 81.3 0.14 16.4 11.0 3. The culture medium of 2 is high-
Processing 2.8 13.0 66.9 0.12 21.7 16.4

As shown in Table 1, the contents of crude fat, crude protein, carbohydrate, and water-soluble dietary fiber were increased by high-pressure emulsification after lactic acid fermentation. The increase in these nutrients was expected to affect the growth and activity of the Trichoderma reesei inoculated after the high pressure emulsification treatment in the present invention.

Further, the change of the culture of lactic acid bacteria by the high-pressure emulsification treatment was confirmed by a microscope. FIG. 2 is a photomicrograph of the ginseng extract before and after the culture of lactic acid bacteria after high-pressure emulsification treatment in the manufacturing method according to the present invention. In Fig. 2, it was confirmed that the lactic acid bacterium cells were disrupted by high-pressure emulsification treatment.

Experimental Example  2: High performance liquid chromatography  Of the extract used Gin Senocide  analysis

In order to measure the content of the ginsenoside components contained in the ginseng extract and the fermented ginseng extract prepared in Comparative Examples and Production Examples, high performance liquid chromatography (UPLC, Waters) was used. The experimental procedure for measuring the content of ginsenosides is described in detail below.

First, 1 g of each of the ginseng extract of Comparative Example and the fermented ginseng extract of Preparation Example was quantitatively put in a 50 mL mass flask, and then dissolved in ionized water. The solution was passed through a C ₁₈ cartridge (Sep-Pak Plus, WAT020515, Waters, USA) in which the solution was activated, washed with water and 30% (v / v) methanol, eluted with 20 mL of methanol, After concentration under reduced pressure, the residue was dissolved in 2 mL of methanol and used for analysis. As a standard, ginsenosides Rb1, Rd, Rg3-R and Rk1 were dissolved in methanol at a concentration of 1 mg / ml to prepare a stock solution of 1.0, 0.1 and 0.01 mg / ml, respectively, Solution to prepare a calibration curve. The analysis conditions are shown in Table 2 below, and the gradient table of the mobile phase is shown in Table 3 below.

Analysis conditions of ginsenoside UPLC analysis conditions  Flow rate 0.6 mL / min  Injection Volume 2 μl  Analyzing Temperature 10 min  Column Temperature 30 ℃  Column ACQUITY UPLC BEH C ₁₈
(1.7 탆, 2.1 횞 50 mm)  Mobile Phase A: water, B: Acetonitrile

Gradient Table Gradient Table Time (min) Flow rate A (%) B (%) Curve One 0 0.6 80 20 2 0.97 0.6 80 20 7 3 4.31 0.6 68 32 7 4 5.97 0.6 50 50 6 5 7.64 0.6 35 65 6 6 7.86 0.6 10 90 6 7 8.98 0.6 10 90 6 8 9.20 0.6 80 20 6 9 10.0 0.6 80 20 6

The content of ginsenoside components contained in the samples obtained in each step of the production of the fermented ginseng extract according to the present invention was measured through the above-described ginsenoside content measuring method, and the results are shown in Table 4 below.

Change in content of ginsenosides
Manufacturing stage  Ginsenoside (mg / g)  Ginsenoside Rb1 Ginsenoside
Rd Ginsenoside
Rg3-R Ginsenoside
Rk1  1. Ginseng Extract 10.51 2.53 0.92 1.14 2. Ginseng extract as a lactic acid bacterium
The cultured medium 8.50 2.78 1.32 1.31 3. The culture medium of 2 was subjected to high pressure emulsification
Trictor without processing
Culture medium cultured in Lysei 3.15 5.19 1.31 1.54 4. The culture medium of 2 was subjected to high pressure emulsification
After treatment,
Lt; RTI ID = 0.0 > 2.23 8.12 2.13 3.02 5. The culture of 4 was heat-treated
Ginseng fermentation product 1.53 8.23 3.42 4.19

As shown in Table 4, the content of ginsenosides of the culture solution (2) in which the ginseng extract was cultured with Lactobacillus perfumant was higher than that of the ginseng extract (1) not cultured with lactic acid bacteria, , And the Rk1 content was slightly increased. In addition, the culture solution (3) in which the ginseng extract was cultivated with Lactobacillus fermentum and cultured with Trichoderma reesei without high pressure emulsification, only increased the content of ginsenoside Rd.

However, the content of ginsenosides Rd, Rg3-R and Rk1 was significantly increased in the culture solution (4) obtained by culturing the ginseng extract with Lactobacillus fermentum and high-pressure emulsification treatment with Trichoderma reesei. In addition, the content of ginsenosides Rd, Rg3-R, and Rk1 was further increased in the ginseng fermented product (5) subjected to further heat treatment in this case, while the ginsenoside Rb1 was remarkably decreased. The change in the content of gincenoside in the stepwise manner is shown graphically in Fig.

In conclusion, it was difficult to increase the content of ginsenosides Rd, Rg3-R, and Rk1 when cultured with Trichoderma reesei without culturing the ginseng extract only with lactic acid bacteria or high pressure emulsification, but the ginseng extract was cultured with lactic acid bacteria, After the emulsification treatment, the content of ginsenosides Rd, Rg3-R and Rk1 could be increased when cultured with Trichoderma reesei. Further, the content of ginsenosides Rd, Rg3-R and Rk1 could be further increased by an additional heat treatment.

Claims (7)

A process for preparing a fermented ginseng extract having increased contents of ginsenosides Rd, Rg3-r and Rk1 comprising the steps of:
a) preparing ginseng extract from ginseng;
b) fermenting the ginseng extract with lactic acid bacteria;
c) destroying the lactic acid bacteria cells by subjecting the fermented lactic acid bacteria to high-pressure emulsification; And
d) Inoculating and culturing Trichoderma reesei in the high-pressure emulsified lactic acid bacteria fermented product to obtain a fermented ginseng extract.
The method according to claim 1, wherein the lactic acid bacterium is Lactobacillus fermentum .
The method according to claim 1, wherein the high-pressure emulsification treatment in step c) is performed 2 to 4 times at 1300 to 1600 bar.
[2] The method according to claim 1, wherein the culturing of Trichoderma reesei in step d) is carried out at 25 to 30 DEG C for 3 to 5 days.
The method according to claim 1, further comprising the step of performing heat treatment at 110 to 125 ° C for 20 to 40 minutes after the step d).
The method according to claim 1, wherein the content of ginsenosides Rd, Rg3-r, and Rk1 in the fermented ginseng extract of step d) is increased by at least two times as compared to the ginseng extract of step a).
A fermented ginseng extract having increased contents of ginsenosides Rd, Rg3-r and Rk1, produced by the process according to any one of claims 1 to 6.

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