KR20030085615A - Manufacturing method of fermented alcoholic drink using medicinal and eatable plants having organic germanium and fermented alcoholic drink obtained therefrom - Google Patents

Abstract

PURPOSE: A manufacturing method of a fermented alcoholic drink using medicinal and eatable plants having organic germanium and the fermented alcoholic drink produced thereby are provided, thereby manufacturing the fermented alcoholic drink containing a large amount of saponin, organic germanium and ethanol contents. CONSTITUTION: A manufacturing method of a fermented alcoholic drink using medicinal and eatable plants having organic germanium comprises the steps of: (1) culturing tissues or seeds of medicinal and eatable plants; (2) adding organic and inorganic germanium into the cultivated tissues or seeds of medicinal and eatable plants and culturing them for 1 to 90 days; (3) adding 10 to 40%(w/v) of carbohydrate or sugar as a carbon source into the cultivated tissues or seeds of medicinal and eatable plants; (4) inoculating 0.1 to 10%(w/v) of yeast cultured in a YPD medium to the cultivated tissues or seeds of medicinal and eatable plants and culturing them for 1 to 10 days; (5) inoculating 0.5 to 50 wt.% of the first cultivated medium into the cultivated tissues or seeds of medicinal and eatable plants and alcohol-fermenting the mixture for 5 to 30 days; and (6) filtering and sterilizing the second cultivated tissues or seeds of medicinal and eatable plants, wherein tissues or seeds of medicinal and eatable plants are natural or cultivated Panax ginseng radix alba, Panax ginseng C.A. Meyer., Bupleurum falcatum radix, Rehmanniae Radix, Cortex Acanthopanacis, garlic, Carthamus tinctorius L, and red beet.

Description

Manufacturing method of fermented alcoholic beverage using medicinal and edible plants containing a large amount of organic germanium and fermented wine obtained therefrom {Manufacturing method of fermented alcoholic drink using medicinal and eatable plants having organic germanium and fermented alcoholic drink obtained therefrom}

The present invention relates to a method for producing functional fermented wine using medicinal and edible plants containing a large amount of organic germanium and fermented wine obtained therefrom. More specifically, the present invention relates to a method for preparing a functional fermented wine and a fermented wine obtained therefrom using medicinal and edible culture plants containing a large amount of organic germanium known as physiologically active substances.

Because of its high physiological and medical activity, organic germanium (bis-carboxyl germanium sesquioxide; molecular formula GeCH ₂ CH ₂ COOH) ₂ O ₃ ; The study of Ge-132) has been the focus of scholars for over 30 years. Natural Ge-132 extracted from plants has been reported to have superior anticancer effects than inorganic germanium and synthesized organic germanium (Jao et al., Dis. Col. & Rect., 33: 99-104,1990). Many herbal medicines such as ginseng, ganoderma and aloe are known to contain large amounts of germanium, and are known to absorb inorganic germanium in the soil and store it in the form of organic germanium in plants (Kwon et al., Korean J. crop sci. , 41 (6): 729-735, 1996).

On the other hand, in increasing the content of germanium in the plant by the traditional cultivation method, soil cultivation has a disadvantage that there are a lot of limitations such as a wide area of the treated area and climatic loss. Therefore, cultured ginseng containing a high concentration of germanium could be obtained by utilizing in-vehicle mass culture techniques of callus, hairy root and adventitious root obtained by using plant tissue culture technology.

An object of the present invention is to provide a method for producing functional fermented wine using medicinal and edible plants containing a large amount of organic germanium.

Another object of the present invention is to provide a fermented wine obtained by the method of producing a functional fermented wine using a medicinal and edible plant containing a large amount of organic germanium.

1 is a graph showing the change in saponin content and elution yield of cultured ginseng strain according to the fermentation period in the present invention.

Figure 2 is a graph showing the change in germanium content and germanium elution yield of cultured ginseng strain according to the fermentation period in the present invention.

Figure 3 is a saponin chromatogram of cultured three weeks obtained according to the present invention.

Method for producing a functional fermented wine using a medicinal and edible plant containing a large amount of organic germanium according to the present invention, in the fermentation of fermented raw materials containing a carbon source to produce a fermented wine, (1) tissue or seed of the medicinal plant or edible plant A culture plant material preparation step of preparing a culture plant material selected from the group consisting of callus, hairy root, adventitious root, or a mixture of two or more thereof, by culturing the embryo; (2) a germanium treatment step of adding organic and inorganic germanium to the culture plants obtained in the culture plant material preparation step, and further incubating for 1 to 90 days; (3) a carbohydrate or sugar as a carbon source to the culture plant material treated in the germanium treatment step in a quantity corresponding to 10 to 40% (v / w) of the culture three raw materials; (4) YPD solid medium (Yeast extract 2-20g / L, Difco peptone 4-40g / L, Dextrose 4-40g / L, Daikon 15-25g) in yeast as fermented strain to the cultured three raw materials added in the above step / l) fermentation broth obtained by culturing in YPD liquid medium after the first subculture at 20 to 35 ℃ for 24 to 72 hours corresponds to 0.1 to 10% (w / v) of the above-mentioned cultured three raw materials Inoculated in an amount to a single stage fermentation step of incubating for 1 to 10 days; (5) a two-stage fermentation step of inoculating the fermentation broth obtained in the one-stage fermentation step in an amount of 0.5 to 50% by weight to the culture three raw materials in the preservation step, alcohol fermentation for 5 to 30 days; And (6) a post-treatment step of packing the supernatant by filtration and sterilizing the fermentation broth obtained after the two-stage fermentation.

A raw material processing step of processing the culture plant material obtained in the culture plant material preparation step between the germanium treatment step and the step of the preparation step into powder, juice or hot water extract may be further included.

Between the two-stage fermentation step and the post-treatment step may further include a low-temperature politics step to leave the fermentation broth obtained in the two-stage fermentation step at a low temperature of 15 ℃ or less.

In the culture plant material preparation step, the tissues or seeds of the culture plants are medicinal plants, such as ginseng, camphor ginseng, wild ginseng, shiho, jihwang, or golpi, and edible plants, such as garlic, safflower, and red beet. Tissues of tissues or seeds may be used.

As the carbon source used in the preparatory step, sugar, particularly preferably sucrose may be used.

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

Method for producing a functional fermented wine using a medicinal and edible plant containing a large amount of organic germanium according to the present invention, in preparing a fermented wine by fermenting the fermentation raw material containing a carbon source, the culture plant raw material preparation step; Germanium treatment step; Pay-per-step; One-stage fermentation step; Two-stage fermentation step; And a post-processing step.

In the preparation of the culture plant material of step (1), the culture plant material selected from the group consisting of callus, hairy root, adventitious root, or a mixture of two or more thereof is grown by culturing the tissue or seed of the medicinal plant or the edible plant. In this step, by cultivating the medicinal plant or breakfast of the edible plant or cultivation of the seed pear to enable the production of fermented liquor production in large quantities to produce a non-polluting culture plant irrelevant to soil, environment and pesticide contamination.

In the culture plant material preparation step, the tissues or seeds of the culture plants are medicinal plants, such as ginseng, camphor ginseng, wild ginseng, shiho, jihwang, or golpi, and edible plants, such as garlic, safflower, and red beet. Tissues or seed embryos may be used, preferably tissues or seed embryos obtained from 4-6 year old ginseng, 20 year old camphor ginseng, 100 year old wild ginseng and the like.

In the germanium treatment step (2), after the organic germanium is dissolved in water and the inorganic germanium is dissolved in nitric acid, the filter and autoclaving are added to the culture plant obtained in the preparation of the plant material preparation, and further cultured for 1 to 90 days. In addition, the culture plant is a step for sufficiently absorbing germanium to be contained in a large amount in the culture plant in the form of organic germanium so that a large amount of organic germanium is eluted by the subsequent fermentation to be included in the fermentation strain. The bioreactor culture had a longer immersion time than other culture methods, resulting in a higher absorption rate of germanium and an increase in the accumulation amount for each culture period. In addition, depending on the crop, plant growth was accelerated up to 60 mg / L.

The step of (3) is the step of assigning carbohydrates or sugars as a carbon source to the culture plant material obtained in the germanium treatment step in an amount corresponding to 10 to 40% (w / v) of the culture plant material. It functions to increase the alcohol content by supplementing the carbon source required for fermentation to allow sufficient fermentation. When the equivalent amount is included in less than 10% by weight of the culture plant material, there is a problem that the alcohol content in the fermented liquor as a final product is lowered due to insufficient carbon fermentation due to lack of a carbon source, on the contrary When more than 40% (w / v) of the culture plant material, the excessive osmosis of the carbon source will increase the osmotic pressure to suppress the growth and activity of the yeast will cause abnormal fermentation.

As the carbon source used in the preservation step, preferably sugars, particularly preferably sucrose may be used, and sugars in particular serve to promote alcohol fermentation.

In the single-stage fermentation step of (4), the YPD solid medium (Yeast extract 2 to 20 g / L, Difco peptone 4 to 40 g / L, dextrose 4 to 40 g of yeast as a fermentation strain in the cultured plant material obtained in the step 1). / l, Umbrella 15-25g / ℓ) fermented strain obtained by culturing in YPD liquid medium for 24 to 72 hours at 20-35 ℃ for 0.1 to 10% by weight of the plant material Inoculating in an amount corresponding to% and incubating for 1 to 10 days, the step of expanding the culture to increase the number of yeast in the fermentation broth. In this case, as the yeast, Saccharomyces cerevisiae may be preferably used, and the yeast may be understood to be well known to those skilled in the art of handling alcohol fermentation so that it can be easily purchased and used. It is self-evident. The inoculated culture plant material may further include a sterilization step of sterilizing the cultured plant material for 10 to 30 minutes in a temperature range of 100 to 130 ° C. before inoculating the yeast with the cultured plant material. It is apparent that this sterilization process will be understood by those skilled in the art. When the temperature at the time of the first subculture is less than 20 ℃ or exceeds 35 ℃, there is a problem that the culture of the yeast activity is not enough due to the degradation of the yeast, and the incubation time at the first passage If less than 24 hours, the culture time is too short, there may be a problem that the expansion culture of the yeast is not enough, on the contrary, if it exceeds 72 hours, yeast activity will be lowered and alcohol fermentation efficiency will be lowered. In addition, in the case of expansion culture in less than one day in the expansion culture, the expansion culture of yeast is also not enough, if it exceeds 10 days, there may be a problem such as a decrease in productivity, and also unnecessary fermentation such as acetic acid fermentation There may be a problem with the implementation.

In the two-stage fermentation step of (5), the fermentation broth obtained in the first-stage fermentation step is inoculated in the culture plant material obtained in the step of fermentation in an amount of 2 to 50% (v / v) and alcohol for 5 to 30 days. As the fermentation step, the culture plant material is further added to the fermentation broth of the one-stage fermentation step including the yeast that has been expanded and cultured to produce a large amount of fermented liquor. If the inoculation amount of the fermentation broth is less than 2%, there may be a problem that the alcohol fermentation period becomes longer and the economical efficiency is lowered. to be.

After the two-stage fermentation step, the post-treatment step of (6) is performed, and the post-treatment step is a step of packing the supernatant by compressing and sterilizing the fermentation broth obtained after the two-stage fermentation. The compression filtration process, sterilization process and packaging process are all easily understood by those skilled in the art.

In addition, a raw material processing step of processing the culture plant material obtained in the culture plant material preparation step between the germanium treatment step and the step of the preparation step into powder, juice or hot water extract solution may be further included. In this raw material processing step, the cultivated plant material obtained in the cultivated plant material preparation step is liquefied, the dried cultivated plant material is pulverized, or the powder is extracted and liquefied by hot water extraction to effect saponins (ginsenosides). Allow people to elute more effectively into fermented wine.

In addition, between the two-stage fermentation step and the post-treatment step may further include a low-temperature politics step to leave the fermentation broth obtained in the two-stage fermentation step at a low temperature of 15 ℃ or less. The turbidity of the fermented wine obtained by the low temperature politics step is lowered to further increase the commerciality. If the stationary temperature in the low temperature politics step exceeds 15 ℃, there may be a problem that the improvement of turbidity is not enough. However, if turbidity improvement is not required, this low temperature politics step can be omitted.

Hereinafter, preferred embodiments and comparative examples of the present invention will be described.

The following examples are intended to illustrate the invention and should not be understood as limiting the scope of the invention.

Experimental Example 1

Callus Induction

Ginseng root slices (2-8 mm2) were collected and Murashige-Skug added 2,4-D (2,4-dichlorophenoxy acetic acid; 2,4-dichlorophenoxyacetic acid) and 1 to 10 mg / L. Inoculated in (Murashige-Skoog) medium and induced callus by culturing in a dark state, the titration concentration of 2 mg / ℓ was confirmed that the most effective irrespective of oxin type.

Experimental Example 2

Callus proliferation

The effect was almost the same when cultured using MS medium (Murashige and Skoog), SH (Schenk and Hildebrandt) medium, B5 (Gamborg) medium, LP (Quoirin and lepoivre) medium and white medium. There was a difference. Among these media, good results were obtained in MS medium and 3 / 4SH medium. Growth regulators affecting callus growth were confirmed to be the best in 2,4-D 2 mg / L or NAA or BSAA 2 to 7.0 mg / L medium.

Experimental Example 3

Induction of hairy root

Using MS medium, SH medium, B5 medium, LP medium and white medium, etc., the slices of medicinal and edible plants (2 to 8 mm 2) were used. After infection with lyzogenes ( A. rhizogenes ) for 1 to 3 days, the bacteria were removed using an antibiotic such as claforan (claforan), and then passaged to the same solid medium to obtain hairy roots.

Experimental Example 4

Induction of Abdominal Muscle

1-7 mg / l of IBAI ( IBA; Duchefa, Com) or NAA in MS medium, SH medium, B5 medium while subcultured at intervals of 2 to 4 weeks in optimal callus culture medium When transferred to the added medium, the adductor was formed after 3-4 weeks of culture. Hydrogen ion concentration (pH) of the medium was confirmed that the growth of the adventitious root in the range of 5.0 to 6.0. To make a solid medium, 0.2% by weight of gelite or agar was added, and the sugar concentration was found to be good at 3% by weight of adventitious root development and growth.

Examples 1-5 and Controls

As a culture plant obtained in the above experimental example, germanium was added to wild ginseng root muscle 0 mg / L (control), 10 mg / L (Example 1), 30 mg / L (Example 2), 60 mg / L (Example 3) ), 120 mg / L (Example 4), 150 mg / L (Example 5), and further cultured for 5 weeks, and then analyzed the germanium content, productivity and utilization of the dried culture plants, The results are shown in Table 1 below.

division Live weight (g) (G) in building % Growth Absorption amount (㎍ · g ^-1 DW) Productivity (mg · l ^-1 d ^-1 ) Culture medium (mg · l ⁻¹ ) (total 4ℓ) process Use Control 470.0 39.5 18.5 - - - - Example 1 496.8 41.2 19.8 180.6 ± 0.2 50.7 17.1 3.5 Example 2 560.8 43.3 23.0 674.4 ± 6.9 208.6 51.4 12.7 Example 3 565.2 44.5 23.3 1243.0 ± 35.6 395.1 102.7 41.2 Example 4 410.2 34.3 16.0 1745.4 ± 27.5 427.6 154.1 76.3 Example 5 292.3 24.9 10.1 2212.5 ± 25.0 393.5 205.4 103.4 Example 6 190.0 16.7 5.5 2801.7 ± 18.6 334.2 256.8 108.8 * Results were obtained after incubation for 5 weeks in the bioreactor.

Examples 6-10

As a culture plant obtained in the above experimental example, germanium was treated with 30 mg / l of wild ginseng root muscle at 0 days (control group), 1 day (Example 6), 3 days (Example 7), 5 days (Example 8) , 7 days (Example 9) and 9 days (Example 10) measured by the treatment period of the germanium in the culture plants according to the amount of absorption and the change in the minerals are shown in Table 2 below, the results after increasing the live weight for 35 days, Obtained by two step culture.

division Control Example 6 Example 7 Example 8 Example 9 Example 10 Ge (ppm) - 45.21 ± 1.10 67.18 ± 32.57 100.82 ± 32.84 135.14 ± 14.79 187.74 ± 1.29 N (%) 0.698 ± 0.016 0.602 ± 0.037 0.668 ± 0.106 - 0.672 ± 0.06 0.612 P ₂ O ₅ (%) 1.05 ± 0.03 0.91 ± 0.04 0.88 ± 0.29 1.01 ± 0.10 0.86 ± 0.01 0.81 ± 0.01 K (%) 5.88 ± 0.24 7.43 ± 0.06 7.020 ± 0.23 7.00 ± 0.14 6.2 ± 0.28 7.14 ± 0.17 Mg (%) 0.29 ± 0.018 0.31 ± 0.021 0.35 ± 0.008 0.34 ± 0.00 0.28 ± 0.018 0.33 ± 0.006 Ca (%) 0.57 ± 0.007 0.37 ± 0.009 0.39 ± 0.012 0.41 ± 0.015 0.44 ± 0.011 0.47 ± 0.022 Si (ppm) 167 154.6 ± 24.8 138.3 ± 6.5 122.4 ± 0.9 113.9 ± 4.3 110.8 ± 6.1 Fe (ppm) 208.2 ± 9.6 201.7 ± 2.2 186.1 ± 11.3 205.0 ± 21.4 215.5 ± 11.5 195.3 ± 0.0 Mn (ppm) 772.6 ± 36.6 946.2 ± 66.0 1071.0 ± 124.3 1052.0 ± 5.7 791.0 ± 59.4 948.2 ± 17.8 Cu (ppm) 70.89 ± 4.0 56.24 ± 0.0 36.36 ± 3.6 35.78 ± 1.6 41.70 ± 0.3 56.67 ± 2.1 Na (ppm) 2730.6 ± 25.8 1184.6 ± 150.1 1253.7 ± 143.1 861.58 ± 282.8 1546.1 ± 275.1 1132.5 ± 43.9 Zn (ppm) 137.8 ± 2.8 132.3 ± 0.5 411.37 ± 0.8 117.9 ± 14.5 131.89 ± 3.0 128.51 ± 0.8

Example 11

First, Saccharomyces cerevisiae as a yeast for use in single-stage fermentation for use as a main hair was first passaged at 30 ° C. for 2 days in a solid medium, and then cultured in a YPD liquid medium. Subsequently, sucrose 25% (w / v) as a carbon source was added to 600 ml of a dispersion obtained by dispersing 12 g of cultivated ginseng roots in water as a culture plant for main culture, and then put it in a 1 L flask, and then at 121 ° C. After sterilization for 20 minutes, 6 ml (1% (v / v)) of the subcultured yeast was inoculated, and cultured for 3 days to use as a mother. The obtained seedlings were inoculated in an amount of 3% (v / v), and the change in saponin content and elution yield of cultured ginseng strains during the fermentation period of 1 to 15 days was measured, and the results are shown in FIG. 1. .

As a result, as shown in Figure 1, the longer the fermentation period was confirmed that the saponin content as well as saponin dissolution rate in the fermented ginseng three weeks constant increases.

Example 12

Except for increasing the fermentation period to 1 to 30 days was carried out in the same manner as in Example 11, and the change in the germanium content and germanium elution yield of cultured ginseng strain according to the fermentation period was measured, the results are shown in Figure 2 Indicated. In addition, saponin chromatogram is shown in FIG. 3.

As a result, as shown in Figure 2, it was confirmed that as the fermentation period is longer, the organic germanium content as well as the organic germanium dissolution rate in the fermented acidic liquor three weeks. In addition, as shown in Figure 3 it was confirmed that the extraction of the active ingredient saponin is also effective.

As a result of the synthesis of the above embodiments, the fermented liquor prepared according to the present invention and the fermented liquor obtained therefrom exhibits sufficient saponin content, organic germanium content, etc., compared to the conventional cultivated plant, but rather ethanol content, etc. It was confirmed that functional fermented strains of culture plants appearing higher were obtained.

Therefore, according to the present invention, a method for producing a functional fermented strain of cultured plants showing sufficient saponin content, organic germanium content, etc., but having a higher ethanol content and the like compared to the conventionally grown cultivated plant, and the functional fermented strain obtained therefrom It is effective to provide.

Although the present invention has been described in detail only with respect to the described embodiments, it will be apparent to those skilled in the art that various modifications and variations are possible within the technical scope of the present invention, and such modifications and modifications are within the scope of the appended claims.

Claims (6)

In preparing fermented liquor by fermenting a fermentation raw material containing a carbon source, (1) cultivating the plant material raw material to prepare a culture plant material selected from the group consisting of cultivated callus, hairy root, adventitious root or a mixture of two or more thereof by culturing the tissue or seed of the medicinal plant or edible plant; (2) a germanium treatment step of adding organic and inorganic germanium to the culture plants obtained in the culture plant material preparation step, and further incubating for 1 to 90 days; (3) a carbohydrate or sugar as a carbon source to the culture plant material treated in the germanium treatment step in a quantity corresponding to 10 to 40% (v / w) of the culture three raw materials; (4) YPD solid medium (Yeast extract 2-20g / L, Difco peptone 4-40g / L, Dextrose 4-40g / L, Daikon 15-25g) in yeast as fermented strain to the cultured three raw materials added in the above step / l) fermentation broth obtained by culturing in YPD liquid medium after the first subculture at 20 to 35 ℃ for 24 to 72 hours corresponds to 0.1 to 10% (w / v) of the above-mentioned cultured three raw materials Inoculated in an amount to a single stage fermentation step of incubating for 1 to 10 days; (5) a two-stage fermentation step of inoculating the fermentation broth obtained in the one-stage fermentation step in an amount of 0.5 to 50% by weight to the culture three raw materials in the preservation step, alcohol fermentation for 5 to 30 days; And (6) a post-treatment step of compressing and sterilizing the fermentation broth obtained after the two-stage fermentation to package the supernatant; Method for producing functional fermented wine using medicinal and edible plants containing a large amount of organic germanium, characterized in that made. The method of claim 1, Between the germanium treatment step and the step of the step of preparing the culture plant material obtained in the culture plant material preparation step further comprises a raw material processing step of processing into a powder, juice or hot water extract containing a large amount of the organic germanium Method for producing functional fermented wine using medicinal and edible plants. The method of claim 1, Medicinal containing a large amount of the organic germanium, characterized in that the fermentation broth obtained in the two-stage fermentation step between the two-stage fermentation step and the post-treatment step further comprises a low temperature politics step to stand at a low temperature of 15 ℃ or less. And a method of producing functional fermented wine using an edible plant. The method of claim 1, In the culture plant material preparation step, the tissues or seeds of the culture plants are medicinal plants, such as ginseng, camphor ginseng, wild ginseng, shiho, jihwang, or golpi, and edible plants, such as garlic, safflower, and red beet. Method for producing a functional fermented wine using medicinal and edible plants containing a large amount of organic germanium, characterized in that the tissue or seed embryo. The method of claim 1, Method for producing functional fermented wine using medicinal and edible plants containing a large amount of organo germanium, characterized in that sugar, particularly preferably sucrose, is used as the carbon source used in the preservation step. Functional fermented wine obtained according to any one of claims 1 to 5.