KR20130107940A - Method of producing fermented red ginseng - Google Patents

Abstract

The present invention relates to a method for preparing fermented red ginseng, specifically, step-fermented red ginseng powder or red ginseng slices with Bacillus sp. And Lactobacillus sp. And it relates to a method for producing fermented red ginseng containing a high concentration of 20S-protopanaxatriol.
According to the present invention, the ginsenoid component of red ginseng is subjected to two stages of fermentation of Bacillus strain and Lactobacillus strain, 20 (S) -protopanaxadiol 20-O-β-D-glucopyranoside, 20 It can be efficiently converted to useful components such as (S) -protopanaxatriol, and this active ingredient can be produced in high concentration to produce fermented red ginseng to improve and enhance kidney function.

Description

Method of producing fermented red ginseng {Method of producing fermented red ginseng}

The present invention relates to a method for preparing fermented red ginseng, specifically, step-fermented red ginseng powder or red ginseng slices with Bacillus sp. And Lactobacillus sp. And it relates to a method for producing fermented red ginseng containing a high concentration of 20S-protopanaxatriol.

Red ginseng is called Panax ginseng CA Meyer, or red ginseng, which is cooked and dried to produce red ginseng.

Because it contains less than 15% of moisture, it is easy to store, and it contains various active ingredients, and it is used as a medicine or health food, and it is a health functional food that Korean people love most.

The main components of red ginseng are glycosides, ginseng fragrances, polyacetylene compounds, nitrogen-containing compounds, flavonoids, vitamins, trace elements, enzymes, antioxidants, organic acids and amino acids. Ginsenosides, such as Rh2, Rs1 and Rs2, which do not exist, are contained and are preferred.

Various pharmacological effects can be expected only with the ingredients of red ginseng itself, but as the interest in the health of modern people increases, there is a demand for red ginseng products that are more effective and endowed with various functionalities. Accordingly, various red ginseng products are developed. have.

On the other hand, ginsenosides contained in ginseng are hardly modified by digestive enzymes, and 20 (S) -protopanaxadiol 20-O-β-D-glucopyranoside (20 (S) is caused by intestinal bacteria. ) -protopanaxadiol 20-O-β-D-glucopyranoside) and 20 (S) -protopanaxatriol (20 (S) -protopanaxatriol) and then digested. 20 (S) -protopanaxadiol 20-O-β-D-glucopyranoside exhibits tumor growth inhibiting activity, and 20 (S) -protopanaxatriol enhances immunity. By the modified material exhibits excellent physiological activity, because the composition and activity of enterobacteria differs from person to person, the conversion efficiency of these substances also appears differently, even if the same intake of red ginseng, the effect is different for each person.

Thus, the ginseng fermented with enterobacteria, ginsenosides intrinsic to ginseng was 20 (S) -protopanaxadiol 20-O-β-D-glucopyranoside, 20 (S) -protopanaxatriol By converting to a high bioactive material such as, etc., techniques for enabling everyone to see the effects of ginseng have been studied in various ways.

The present inventors are interested in converting ginsenosides contained in red ginseng into effective ingredients for the treatment or amelioration of certain diseases through microbial fermentation, and can convert ginsenosides inherent in red ginseng into substances having excellent physiological activity. High concentrations of useful components such as 20 (S) -protopanaxadiol 20-O-β-D-glucopyranoside, 20 (S) -protopanaxatriol, etc. Efforts have been made to develop fermentation methods for producing fermented red ginseng.

Therefore, the main object of the present invention is a high concentration of physiologically active substances such as 20 (S) -protopanaxadiol 20-O-β-D-glucopyranoside, 20 (S) -protopanaxatriol through microbial fermentation. To provide a method for preparing fermented red ginseng containing.

According to one embodiment of the present invention, the present invention comprises: a) adding 50 to 150 parts by weight of water to 100 parts by weight of red ginseng powder or red ginseng slices, and adding 2 to 20 parts by weight of Bacillus strain culture solution at 48 to 35 Fermentation for 120 hours; b) heating the fermented product produced in step a) to 90 to 120 ° C. for 5 to 60 minutes; And c) adding 100 to 300 parts by weight of water and 2 to 20 parts by weight of Lactobacillus strain culture medium to 100 parts by weight of the fermented product sterilized in step b), and fermenting at 35 to 40 ° C. for 48 to 120 hours. It provides a method for producing fermented red ginseng for improving and strengthening kidney function containing 20S-protopanaxadiol 20-O-β-D-glucopyranoside and 20S-protopanaxatriol.

In the fermented red ginseng manufacturing method of the present invention, it is preferable that the filtrate of the fermentation product produced in step c) is lyophilized or hot air dried.

According to another aspect of the present invention, the present invention provides an improvement in renal function containing 20S-protopanaxadiol 20-O-β-D-glucopyranoside and 20S-protopanaxatriol prepared by the above method. Provide fermented red ginseng for fortification.

In the method of manufacturing fermented red ginseng of the present invention, red ginseng used as a raw material is steamed and dried to be processed so that the water content is about 15% or less, and there are Cheonsam, Jisam, Yangsam, Jesamsam, and Misam which are classified according to quality. It is not limited only to these. Moreover, these red ginseng can be used individually or in combination.

The ginseng used as a raw material of red ginseng refers to a plant of the genus Panax containing glycoside as a main ingredient, and ginseng (Korea ginseng; Panax ginseng CAMeyer), Sanchi ginseng (notoginseng (burk.) FHChen), American ginseng; Panax quinquefolium L. Ha et Grushv.), But is not limited to these. Moreover, these ginseng can be used individually or in combination.

The red ginseng can be pulverized and used as red ginseng powder, and the red ginseng can be cut into a certain thickness to be used as red ginseng slices, and in the case of red ginseng powder, it is preferable to use about 100 to 400 mesh crushed. It is preferable to use what cut | disconnected about 1-10 mm. More preferably, the red ginseng powder may be crushed at about 200 mesh, and the red ginseng slice may be cut at about 2 mm.

Bacillus genus strains include Bacillus Richenomis ( Bacillus licheniformis ), Bacillus amiriquifaciens ( Bacillus amyloliquefaciens ), Bacillus subtilis subtilis), Bacillus NATO (Bacillus natto ), Bacillus polymisa polymyxa ), but not limited thereto. These Bacillus strains can also be used alone or in combination.

LB (Luria Bertani) medium, nutrient medium, potato glucose medium or soybean medium may be used for culturing the Bacillus strain, and may be cultured at stationary or shaken at a temperature of 35 to 40 ° C. In the present invention, by incubating the shaken at about 37 ℃ for 2 days can be added directly to the culture medium of the number of the cells of about 1.0 × 10 ⁵ ~ 1.0 × 10 ⁹ CFU / g, only the cells can be recovered and then suspended suspension with water can be added. .

To the Lactobacillus genus Lactobacillus strains Planta Rum (Lactobacillus plantarum), Lactobacillus four K (Lactobacillus sakei ), Lactobacillus fermentum ), Lactobacilus rhamnose ), Lactobacillus gasseri ), Lactobacilus mali ), but not limited to these. Furthermore, these Lactobacillus strains can be used alone or in combination.

It is preferable to use MRS medium for culturing the Lactobacillus strain, it can be left standing or shaking culture at a temperature of 35 ~ 40 ℃. In the present invention, by incubating the shaken at about 37 ℃ for 2 days can be added directly to the culture medium of the number of the cells of about 1.0 × 10 ⁵ ~ 1.0 × 10 ⁹ CFU / g, only the cells can be recovered and then suspended suspension with water can be added. .

In the fermented red ginseng manufacturing method of the present invention, step a) is a step of first fermentation with Bacillus sp. Strain, adding 0.5 to 1.5 times the weight of water to supply water to red ginseng, and adding Bacillus sp. Strain culture solution thereto. It is characterized by forming conditions under which Bacillus strains can be easily fermented. Since red ginseng has low water content, fermentation is rarely performed or fermentation efficiency is very low. Therefore, the red ginseng absorbs water by supplying an appropriate amount of water, and provides an environment for the growth and metabolism of Bacillus strains on the surface of the red ginseng powder or red ginseng slices. The addition of such amounts of water forms a thick mixture.

Step b) in the fermented red ginseng production method of the present invention is to control the growth and activity of the Bacillus strain in the red ginseng fermented with Bacillus strains to facilitate the fermentation of Lactobacillus strains. have. If the sterilization under the above conditions may not be complete sterilization, due to the thermal shock may lower the activity of the genus Bacillus strains. When heated to a temperature lower than the above range and for a short time, it is difficult to expect the inhibitory effect of the strain of the genus Bacillus, it is not preferable because the components produced through the primary fermentation may be modified when heated to a high temperature and a long time. After heating, it is preferable to cool enough so as not to inhibit the activity of the Lactobacillus strain in the second fermentation.

In the fermented red ginseng production method of the present invention, the step c) is a step of second fermentation with a strain of the genus Lactobacillus, further adding 1 to 3 times the weight of water to the fermented product by primary fermentation, and lactose By adding a Bacillus strain culture medium, it is characterized by forming conditions under which the Lactobacillus strain can be easily fermented. Since the first fermented fermentation is not immersed in water, it is not suitable for growth of Lactobacillus strains that grow well in a low oxygen environment. Therefore, by adding more water to the primary fermentation, the primary fermentation is immersed in water, and when Lactobacillus strain culture medium is added thereto, the growth and fermentation of the strain Lactobacillus can be easily performed.

The fermentation product obtained through the secondary fermentation may be sterilized and commercialized or ingested as it is, but it is preferable to prepare the fermentation broth by filtering the fermentation product and then freeze-drying or hot-air drying to form a concentrated active ingredient.

In addition, the active ingredient can be concentrated by extracting the fermented product or fermented broth from secondary fermentation with alcohol, water or a mixed solution thereof. At this time, the obtained extract may be freeze-dried or hot-air dried to further concentrate by removing alcohol or water in the extract.

According to the present invention, the ginsenoid component of red ginseng is subjected to two stages of fermentation of Bacillus strain and Lactobacillus strain, 20 (S) -protopanaxadiol 20-O-β-D-glucopyranoside, 20 It can be efficiently converted to useful components such as (S) -protopanaxatriol, and this active ingredient can be produced in high concentration to produce fermented red ginseng to improve and enhance kidney function.

1 is a block diagram showing an embodiment of the fermented red ginseng manufacturing method of the present invention.

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

Example  One. Bacillus  Strain Selection and Identification

Cheonggukjang is suspended in sterile water, plated in LB solid medium and incubated at 37 ° C. for about 2 days. Strains are isolated from each colony formed in the medium, and gram staining and spore staining are used to isolate strains that form spores with Gram-positive bacilli.

Each strain isolated in the LB liquid medium containing red ginseng powder was inoculated, incubated at 37 ° C. for about 2 days, and the culture solution from which the cells were removed was analyzed by high performance liquid chromatography (HPLC).

As a result, strains with excellent ginsenoside conversion ability of red ginseng are selected.

The 16S rRNA gene sequences of each selected strain are analyzed using conventional PCR (polymerase chain reaction) and sequencing methods, and compared with the NCBI database.

Example  2. Bacillus  Red Ginseng Fermentation Using Strains

2-1. Bacillus  Strain Culture Preparation

Bacillus strains selected in Example 1 were inoculated in LB liquid medium and cultured at 37 ° C. for about 2 days to prepare a strain culture.

2-2. Red Ginseng Fermentation

1 kg of water is added to 1 kg of red ginseng powder having a thickness of about 100 to 400 mesh or red ginseng having a thickness of about 2 mm, and mixed evenly while spraying 100 g of the culture solution of 2-1.

After fermentation for about 4 days at 37 ℃.

Example  3. Lactobacillus  Strain Selection and Identification

Kimchi or animal small intestine samples are cut and suspended in sterile water, plated in MRS solid medium containing 0.5% CaCO ₃ and incubated at 37 ° C. for about 1 day. Of each colony formed in the medium, strains are isolated from the colonies that have a transparent ring around them, and gram positive and catalase negative strains are selected by examining gram staining and catalase activity.

Each strain isolated to the MRS liquid medium containing the fermentation broth obtained by filtering and sterilizing the fermentation product of Example 2 was inoculated and incubated at 37 ° C. for about 2 days, and the culture medium from which the cells were removed was HPLC (high performance liquid chromatography). Analyze with

As a result of the analysis, strains having excellent production ability of ginsenoside converting materials such as 20S-protopanaxadiol 20-O-β-D-glucopyranoside and 20S-protopanaxatriol are selected.

The 16S rRNA gene sequences of each selected strain are analyzed using conventional PCR (polymerase chain reaction) and sequencing methods, and compared with the NCBI database.

Example  4. Fermented Red Ginseng Manufacture

4-1. Bacillus  Strain Culture Preparation

Bacillus strains selected in Example 1 were inoculated in LB liquid medium and cultured at 37 ° C. for about 2 days to prepare a strain culture.

4-2. Lactobacillus  Strain Culture Preparation

The strain Lactobacillus selected in Example 3 was inoculated in MRS liquid medium and cultured at 37 ° C. for about 1 day to prepare a strain culture solution.

4-3. Fermentation

1 kg of water is added to 1 kg of red ginseng powder of about 100 mesh or red ginseng slices of about 2 mm thickness, and then mixed evenly while spraying 100 g of Bacillus strain culture of 4-1, and then fermented at 37 ° C. for about 4 days. do.

The resulting fermentation is heated to 100 ° C. for 20 minutes.

2 kg of water is added to the sterilized fermentation product, and 100 g of the Lactobacillus strain culture solution of 4-2 is evenly mixed therein, followed by fermentation at 37 ° C. for about 4 days.

4-4. concentration

The fermented product of 4-4 is filtered to obtain a fermentation broth, a part of which is lyophilized or hot air dried.

Or 4-4 fermented product is extracted step by step using 95% ethanol, 75% ethanol, 55% ethanol, water, and the obtained extracts are mixed to remove ethanol and lyophilized or hot air dried.

Claims (3)

a) adding 50 to 150 parts by weight of water to 100 parts by weight of red ginseng powder or red ginseng slices, and adding 2 to 20 parts by weight of Bacillus strain culture to ferment for 48 to 120 hours at 35 to 40 ° C;
b) sterilizing the fermented product produced in step a) at 90 to 120 ° C. for 5 to 60 minutes; And
c) adding 100 to 300 parts by weight of water and 2 to 20 parts by weight of Lactobacillus strain culture medium to 100 parts by weight of the fermented product sterilized in step b) and fermenting at 35 to 40 ° C. for 48 to 120 hours.
20S-protopanaxadiol A method for producing fermented red ginseng for improving and enhancing kidney function, which contains 20-O-β-D-glucopyranoside and 20S-protopanaxatriol. The method of claim 1,
20S-protopanaxadiol 20-O-β-D-glucopyranoside and 20S-protopanaxatriol, characterized in that the filtrate of the fermentation product produced in step c) is lyophilized or hot-air dried. Fermented red ginseng manufacturing method for improving and strengthening the kidney function. A fermented red ginseng for improving and strengthening kidney function, which contains 20S-protopanaxadiol 20-O-β-D-glucopyranoside and 20S-protopanaxatriol prepared by the method of claim 1 or 2.

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