KR20130037154A

KR20130037154A - Mushroom extract manufacturing method of anti-cancer efficiency

Info

Publication number: KR20130037154A
Application number: KR1020120078662A
Authority: KR
Inventors: 김용규; 김민선
Original assignee: 김민선; 김용규
Priority date: 2011-10-05
Filing date: 2012-07-19
Publication date: 2013-04-15

Abstract

PURPOSE: A method for preparing a fermentation of mushroom extract is provided to enhance anticancer activity by fermenting mushroom extract and makkoli(Korean rice wine). CONSTITUTION: A method for making makkoli comprises the following steps: a step of washing, dipping, and steaming rice and preparing hard-boiled rice; a step of mixing with seed malt and preparing ipguk(Japanese koji); a step of preparing a crude liquor; and a step of primarily and secondarily fermenting and preparing suldeot(rice wine mash). A method for preparing an fermentation for anticancer comprises: a step of mixing 1L of groundwater, 0.01ml of Ganoderma lucidum extract, 0.01ml of Phellius linteus extract, 0.05ml of Sparassis crispa extract, 0.05ml of Lentinula edodes extract, 0.01ml of Cordycaps sinensis extract, 0.05ml of an Inonotus obliquus extract, 0.1ml of red ginseng extract, and 0.1ml of ginseng extract.

Description

Anti-cancer fermentation method containing mushroom extract {MUSHROOM EXTRACT MANUFACTURING METHOD OF ANTI-CANCER EFFICIENCY}

The present invention relates to a method for producing an anticancer fermentation product containing a mushroom extract, and more specifically, extracts including red ginseng, ginseng, ganoderma lucidum mushroom, situation mushroom, zinnia mushroom, shiitake mushroom and Cordyceps sinensis, and edible mushroom with a certain fermentation product. It relates to a method for producing an anticancer fermentation product comprising a mushroom extract to enhance anticancer activity by fermentation.

As is well known, the representative of the fermented material is liquor, bar, the beneficial enzyme is activated through the fermentation process, look at the example of the main wine Makgeolli, rice wine manufacturing method is glutinous rice, non-rice, barley, flour, etc. It is a Korean sake that is fermented by mixing malt and water. It is also known as Takju, Nongju (農酒), Jaju (회 酒) and Hoeju (灰酒). It is the oldest alcoholic beverage in Korea, and the color is white and turbid like hot water, and the alcohol content is 6-7 degrees.

The manufacturing method of the makgeolli manufacturing method is mainly steamed glutinous rice, non-glutinous rice, barley, wheat flour, etc., and dried water, which is called jiebab, mixed with leaven and water and fermented at a constant temperature without squeezing the sake. In the old general family, the liquor made by mixing yeast with Jiebab is placed on a crock bowl and put on a bowl of 井 -shaped, and then poured into a sieve. )

At this time, a method of manufacturing glutinous rice makgeolli, which is made of glutinous rice as raw materials, is called Dong-dongju, which is left uncooked with rice paste. A good method for producing makgeolli is sweet, sour, bitter, and astringent. It has a rich taste and a cool taste. It has been used as a farmer by relieving the thirst of farmers who sweat and work.

The alcohol content of makgeolli is similar to beer, and makgeolli is a fermented food, rich in functional ingredients such as yeast, protein, sugar, vitamin B2, and choline. Makgeolli manufacturing method and the cardiovascular disease prevention effect, the dietary fiber and protein components of the makgeolli manufacturing method is known to have an anti-cancer effect.

However, since the anticancer effect present in the rice wine manufacturing method itself is very insignificant, the anticancer effect was somewhat insufficient.

The present invention has been made in view of the above-described prior art, anti-cancer by fermenting the extract including red ginseng and ginseng, ganoderma lucidum mushroom, mushrooms, matsutake mushrooms, shiitake mushrooms and Cordyceps sinensis mushrooms and a method for producing rice wine It is an object of the present invention to provide a method for producing an anti-cancer fermented product containing a mushroom extract to enhance the action.

In order to achieve the above object, according to a preferred embodiment of the present invention, the step of semi-precipitating and increasing the steam to form a gourd rice and mixing the final state to prepare the entry, preparing a base liquor (primary hair), primary fermentation step, In the method of manufacturing makgeolli, comprising the step of obtaining the fermentation and the second fermentation, 1 liter of groundwater extracted from 150 meters of rock water as brewing water, 0.01 ml of ganoderma lucidum extract, 0.01 ml of situation mushroom extract, and 0.05 ml of mushroom mushroom extract And, mushroom extract, characterized in that the mixture of shiitake mushroom extract 0.05ml, Cordyceps sinensis extract 0.01ml, and the mushroom extract 0.05ml, red ginseng extract 0.1ml and ginseng extract 0.1ml volumetric mixture An anticancer fermentation method is provided.

Preferably, the mushroom and red ginseng and ginseng extract is separated from Ganoderma lucidum mushroom, situation mushroom, matsutake mushroom, shiitake mushroom, Cordyceps sinensis, tea mushroom, red ginseng, ginseng, respectively, after washing the roots and stems and removing foreign substances, cut to thickness 0.5㎝ And then separately put into separate containers, the drying step of drying at 50 ℃, 12 hours into a hot air precursor room; The dried materials were separately extracted with a mixture of water and ethanol in a ratio of 1: 9, and extracted 1 to 2 times in a range of 65 to 105 ° C. with a reflux extractor, and filtered using a vacuum filter. In addition, there is provided a method for producing an anticancer fermentation product comprising a mushroom extract, wherein the filtrate is prepared through an extraction process of concentrating each 60-25 Brix using a vacuum concentrator.

The anti-cancer fermented product manufacturing method including the mushroom extract according to the present invention can obtain enhanced anti-cancer effect while enjoying the fermented product, which is not comparable as a health drink, and has the advantage of using various pharmacological actions of the mushroom.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail with reference to drawings.

The anticancer fermentation product containing the mushroom extract according to an embodiment of the present invention undergoes a process of replacing the brewing water containing the mushroom extract with excellent anticancer effect in a process including brewing water during the process of preparing the fermentation product. It features.

The material that uses rice as a main ingredient such as makgeolli or Sikhye of representative fermentation is applicable. The raw material of makgeolli is generally rice, and rice is an important raw material for manufacturing makgeolli. The raw material affects the main quality.

General processes for the production of makgeolli are generally semi- and muddy, watery, steamed, ultimately, entry, base wine, primary fermentation, and secondary fermentation.

The manufacturing process of makgeolli first begins with a process of semi and saliva, and the purpose of the semi and saliva is to remove dust and the like adhering to the surface of rice and to absorb an appropriate amount of water. In general, the immersion time of rice depends on the net rice ratio, and the water absorption ratio after draining water is appropriate around 25 ~ 28%.

The cooker is a process of making gourd rice, and the purpose of cooker is to gelatinize the starch with strong steam of 100 ℃ or higher on the rice which absorbed water to facilitate the action of various enzymes. The steaming time is about 40 to 60 minutes, but it is controlled according to the boiler's performance. The steam is increased for about 20 to 30 minutes so that the weight of the rice is increased by 35 to 42% of the rice.

The final station refers to fungi which are used as spawns in the manufacture of the station. Usually, certain molds are bred in millet or rapeseed to contain many spores.

Entry is the artificial breeding of molds (final) after adding starch ingredients. Takju and Yakju's entry is entirely white. This is because unlike the yellow soup (黄 () strong productivity (生酸性) because it prevents the contamination of various bacteria (술) in drunk (main).

Baekkukyun, which is widely used at present, is a kind of mutant strain that is mutated from Blackukkuk. It is called "Aspergillus. Kawachill".

Base liquor refers to the expanded culture of yeast that ferment the base liquor. Base liquor has a higher media concentration (reduced water supply), higher acidity, and lower incubation temperature compared to drunk (main).

Base liquor must contain a large amount of healthy enzymes and acids (hydrangea under citric acid, yeast under liquor: lactic acid). The presence of acid prevents contamination of germs at the base and stage 1, and furthermore, at the beginning of the stage 2 stage (main stage), even when there is the greatest risk of bacterial contamination due to inadequate yeast growth and alcohol production. It serves to prevent.

In order to manufacture a healthy base liquor (base hair), it is necessary to secure a base thread completely separated from the fermentation chamber so that various bacteria are not contaminated and cultivate or purchase a superior yeast. If you use an unhealthy base, there is a risk of dissatisfaction.

In the first fermentation, one-stage immersion refers to immersion of base liquor, entry, and water as raw materials, and is a step of expanding and cultivating yeast necessary for fermentation step by step as a second step of the base liquor manufacturing process.

The purpose of the monolith is the leaching of various enzymes and acids secreted by the entry, the self-glycosylation of the entry, and yeast growth under safe conditions.

First-stage immersion is completed by first adding water, base liquor and entry into the immersion container. The soaking temperature is about 22 ~ 28 ℃, and the soaked water will reach the time of immersion due to the saccharification of the immigration itself and the growth of yeast. The period from 1 st dip to main dip is about 5-6 days. Stir it once or twice a day during this period. The best product temperature during the first stage immersion period is 27-28 ℃.

Second fermentation is a ripening process. When only yeast (grain) is used, the first stage of immersion is the main immersion, and when the immigration is mixed or dedicated, the two stages of immersion is called main immersion. Complete the main immersion by mixing the water supply, saccharifying enzyme (leav, coenzyme, and purified enzyme) and Gwami well in one stage immersion.

This immersion product temperature is the same as the one-stage immersion product temperature. After about 5 hours after the main immersion, the immersion liquid absorbs water (leaked enzyme) enough to become saccharified and alcoholic fermentation.

Thus, the immersion maximally expands and then sinks slightly due to dissolution saccharification and fermentation. After aging, filter the liquor ants to eat.

Yeast is a kind of soup that grinds wheat and roe wheat to form dough, and then naturally breeds molds in the air to produce and secrete various enzymes. Since it has wild yeast, it is a kind of fermenting agent that acts as the mother of base wine. to be.

The brewing water in the production of makgeolli is not only a component of the product, but also a solvent of all ingredients and enzymes during the brewing process, and the trace inorganic components in the water play an important role as nutrients and stimulants of microorganisms that ferment. .

In particular, brewing water, which accounts for more than 80% of the product, has a huge impact on the quality. Therefore, it should be chosen as colorless and transparent, free from blemishes and odors, neutral to weakly alkaline, containing an appropriate amount of active ingredients, and free from harmful microorganisms and harmful components.

As a brewing water for the anticancer fermentation product containing the mushroom extract according to an embodiment of the present invention using the water drawn up from 150 meters underground, 1 day immersion, 1 day empire and room temperature (eg, 33 ℃ to 38 ℃) Final two days.

At this time, the brewing water of the anticancer fermentation product containing the mushroom extract according to an embodiment of the present invention is mixed with a certain amount of mushroom extract in 1 l of water drawn up from 150 meters underground to further enhance the anticancer action.

At this time, the mushroom extract is an extract that further comprises Ganoderma lucidum mushroom, situation mushroom, matsutake mushroom, shiitake mushroom, Cordyceps sinensis, chaga mushroom, red ginseng and ginseng extract.

The Ganoderma lucidum extract 0.01ml is mixed with 1ℓ of water, Ganoderma lucidum is the main medicinal effect in asthma, bronchitis, arthritis, hypertension, cancer treatment, etc. Ganoderma lucidum is divided into a high molecular weight component and a low molecular weight component, Are mainly immunostimulatory proteins and anticancer polysaccharides (or beta-glucans). On the other hand, the low molecular weight components are triterpenoids extracted with organic solvents and have a strong metabolite.

At this time, beta-glucan is a polymer polysaccharide that is not produced at all in the human body. These polysaccharides are more prophylactic to activate immune function than to cure diseases, lower cholesterol levels excessively in the blood, and regulate the amount of sugars that are excessively transferred, which is an essential condition for diabetics. Controlling excess cholesterol or sugar in the blood eventually clears up the blood, which inevitably improves immune function.

Beta-glucan's bio-always effects are effective against cancer, diabetes and adult diseases as well as allergic diseases and atopic dermatitis seen in infants.

At this time, the mushroom extract, including Ganoderma lucidum mushroom, mushroom mushroom, matsutake mushroom, shiitake mushroom, Cordyceps sinensis, chacha mushroom, and the extraction method of red ginseng and ginseng extract will be described below the extraction method.

Example 1: Extract Preparation

1) Drying process

After washing the roots and stems of Ganoderma lucidum mushrooms, situation mushrooms, matsutake mushrooms, shiitake mushrooms, Cordyceps sinensis mushrooms, red mushrooms, red ginseng, and ginseng, remove foreign substances, cut them into 0.5cm thick, and put them in separate containers. Put it in the hot air preheating chamber and let it dry at 50 ℃ for 12 hours.

2) Extraction and Concentration Process

The dried materials were separately extracted with a mixture of water and alcohol in a ratio of 1: 9, and extracted 1-2 times for 6 to 12 hours at a temperature of 65 to 105 ° C. using a reflux filter, followed by filtration using a vacuum filter. The filtrate was concentrated to 60-25 Brix each using a vacuum concentrator to prepare extracts of each material.

Preferably, in order to diversify the extraction concentration, the volume ratio of water and alcohol in the extraction solvent may be concentrated using a mixed solution within a range of 1: 1, 2: 1, 1: 2, 1.5: 0.5, and the like. Do.

On the other hand, the situation mushroom extract is mixed with 0.01 ml per 1 L of water, the situation mushroom is being studied for two kinds of Phellinus linteus and Phellinus baumii. Polysaccharide of the situation mushroom is expressed as an excellent anti-cancer effect, and the yellowish substance shows various pharmacological effects.

This yellow substance is a polyphenol-based pigment compound, and exhibits antioxidant activity, anti-diabetic action, dementia prevention action, antiviral action, anti-inflammatory action, cancer cell death action, platelet aggregation inhibition action and the like.

Looking only at the representative pharmacological action of the situation mushroom, as a stabilization component of the situation mushroom, components such as hispidin (hispidin), bihispidinyl, hypohymine B (hypholomine B) is known, Neil (bihispidinyl), hypolomine B (hypholomine B) is more antioxidant than hispidine.

In addition, the situation mushroom is histidine isolated from the mycelium culture medium of Linteus situation mushroom exhibits the effect of inhibiting PKC-beta to kill cancer cells more strongly than normal cells. Phenoligridin C and phellifuropyranone A in S. mushrooms also have toxic effects on human cancer cell lines.

In this case, the meaning of mixing 0.01 ml of the situation mushroom extract with respect to 1 liter of water means that 1 liter of water and 0.01 ml of the situation mushroom extract are mixed to make a mixed solution.

In addition, the flower mushroom extract is mixed with 0.05ml per 1L of water, the flower mushroom may contain antibacterial and antifungal substances, but sparan (sparan, beta-glucan) is included, so it promotes immune There is action and anticancer action.

Sparan inhibits angiogenesis and cancer metastasis in cancer-expressing mice, and is effective in lung cancer, stomach cancer, colon cancer, breast cancer and prostate cancer in human clinical trials. Sparans induce a variety of immunostimulatory cytokine secretion in human peripheral blood mononuclear cells. Sparan stimulates immunity by secreting TNF-alpha, IL-12, IFN-gamma, GM-CSF in macrophages and dendritic cells.

The shiitake mushroom is mixed with 0.05ml of shiitake mushroom extract per 1ℓ of water, shiitake mushrooms are excellent for preventing various adult diseases, inhibiting cancer cell proliferation, hypertension, diabetes, etc., and the general ingredients contain a lot of protein, fat, sugar .

Vitamins contain vitamins B1, B2 and niacin, but no vitamins A and C. Shiitake mushrooms also contain large amounts of iron that produces hemoglobin in the blood.

In addition, a special ingredient called eridadenine removes cholesterol in the blood, thereby preventing high blood pressure.

In particular, lentinan contained in shiitake mushrooms is involved in anticancer activity by increasing the activity of natural killer cells (NK cell). Lentin is a protein of 27.5 kDa molecular weight isolated from shiitake fruiting bodies. It suppresses the mycelial growth of various fungi.

The Cordyceps sinensis is mixed with 0.01ml of the Cordyceps sinensis extract per 1 liter of water. Cordyceps sinensis is a fungi parasitic and grows in insects rather than plants, and deoxyuridine among the substances contained cordyceps inhibits cancer cell growth. , 2'3'-dideoxyadenosine (2'3-dideoxyadenosine) also inhibits cancer cell growth, hydroxyethyladenosine (hydroxyethyladenosine) also inhibits cancer cell growth, cordycepic acid (cordycepic acid, D-mannitol) is an anticancer agent.

Chaga is a mixture of 0.05 ml of chaga extract per 1 L of water. Chaga is a white fungus that grows in Russia, Europe, Asia, and North America, and does not form fruiting bodies. Form a lump. These chunks are called chaga in Russian.

Among the components of chaga, inotodiol has anticancer and hypoglycemic effects, and hisslon has suppressed growth of cancer cell lines.

The red ginseng and ginseng extract is mixed at a rate of 0.1 ml each for 1 L of water, and the saponin component and non-saponin substance of red ginseng have activity to inhibit the proliferation of various types of cancer cells. The major active ingredients that inhibit the proliferation of cancer cells are G-Rh2, a saponin ingredient unique to red ginseng, and polyacetlen, an important component of fat-soluble fractions (petroleum ether extract), as a non-saponin substance. Red ginseng has a remarkable effect on the activation of natural killer cells in vivo that can kill cancer cells.

Mushrooms are known to have a variety of physiologically important effects, including antibacterial, anticancer and immune system regulation. There are many known substances that show anticancer activity in mushrooms, most of which are polysaccharides such as beta-glucan, lentinan, and lectin.

As discussed above, each mushroom extract has been shown to contain anticancer substances and have anticancer effects through long-term studies around the world, but this is an anticancer action of each mushroom extract, Although it can be proved indirectly that the anticancer makgeolli of the present invention has some effects on the anticancer properties, it does not prove the direct anticancer effect of the fermentation product containing the mushroom extract of the present invention, and it is also necessary to perform a direct experiment. I will.

Example 2: Effect of Anticancer Fermentation and Cisplatin on Anticancer Activity of Mushroom Extract

OBJECTIVES: The effect of anticancer makgeolli on cancer cell toxicity was investigated by measuring the viability of cisplatin and anticancer fermentation into cultured cancer cells.

Experimental Method:

1. Cultivation of Cells

Human lung cancer cells (A549 cells) were distributed to Korea Cell Line Bank and used for the experiment. RPMI 1640 medium (Gibco BRL) containing 10% fetal calf serum, streptomycin (100 U / ml) and penicillin (100 U / ml) was incubated at 37 ° C. in a CO 2 incubator and used for the experiment.

2. Experimental group

The experimental group was divided into anticancer fermentation group, cisplatin addition group, and anticancer fermentation product + cisplatin addition group.

The experiment was incubated for 1 day in lung cancer cells (A549 cells: 2 × 10 ⁵ cells) in a cell culture vessel (96 well plate), the anti-cancer fermentation group added to the culture medium containing the anti-cancer fermentation product prepared in Example 1 ( Anti-cancer fermentation concentration: 0-5 mg / ml), cisplatin added group was added to cisplatin (cisplatinum: Sigma chemical Co, USA) culture solution (cisplatin concentration: 0-5 mg / ml), anti-cancer fermentation + cisplatin added group For example, the cells were cultured in a CO2 incubator for 48 hours after being exchanged with a culture solution added with an anticancer fermentation product and cisplatin (mushroom extract concentration: 0-5 mg / ml, cisplatin concentration: 0-5 mg / ml), and then cell activity (Cell viability). ) Was measured by MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl-tetrazolium bromide) method to evaluate the cellular activity by converting the absorbance of the experimental group to the control group as a ratio. Repeated experiments were carried out and expressed as average values (Table 1).

Experiment result:

Effect of anticancer fermentation and cisplatin on survival of lung cancer cells (A549 cells) Cisplatin Concentration
(Μg / ml) Percentage of cell activity of lung cancer cells (%) Control group Anticancer Fermentation Additive Group Concentration: 1 mg / ml Concentration: 2 mg / ml 0 100 100 100 50 99 99 99 100 96 98 96 200 98 92 87 400 97 88 76 600 91 79 68 800 82 69 65

The cell activity of the control group (addition of cisplatin only) decreased the cell activity of lung cancer cells to 82% at the concentration of cisplatin at 800 μg / ml.

Cellular activity of the anticancer fermentation product of 1 mg / ㎖ of the present invention is reduced the cell activity of lung cancer cells at cisplatin concentration of 200 ㎍ / ㎖ or more, cells of lung cancer cells at cisplatin concentration of 200, 400, 600, 800 ㎍ / ㎖ Activity was 92, 88, 79 and 69%, respectively.

The cell activity of the anticancer fermented 2 mg / ml group was further decreased in the lung cancer cells at the cisplatin concentration of 200 ㎍ / ml or more, the cell activity of the lung cancer cells at the cisplatin concentration of 200, 400, 600, 800 ㎍ / ml 87, 76, 68 and 65% were shown, respectively.

Looking at the above experimental results, it can be seen that the addition of cisplatin and anti-cancer fermentation reduces the cellular activity of lung cancer cells than when adding cisplatin alone. This phenomenon is due to the anticancer action of the polysaccharides contained in mushrooms, ginseng and red ginseng. In more detail, the polysaccharides are not directly anti-cancer activity, and it is thought that cisplatin acts to increase anticancer activity against lung cancer cells.

The method for producing a fermented product using the sulfur hot spring water of the present invention is any well-known method for producing makgeolli, known in the art, specific mushroom extract and ginseng, red ginseng which can enhance anticancer activity in brewing water. It is not limited to a specific brewing method by preparing the makgeolli by mixing the extract.

On the other hand, the anticancer fermentation method manufacturing method containing a mushroom extract according to an embodiment of the present invention is not limited to only the above-described embodiment and various modifications are possible within the scope not departing from the technical gist.

Claims

In the method of manufacturing makgeolli, which comprises the step of semi-precipitating and cooking steam to form gourd rice and mixing the final ingredients, preparing the entry, preparing the base liquor (primary), the first fermentation step, the second fermentation step, and drunking. 1 liter of groundwater extracted from 150 meters of rock bed water, 0.01 ml of ganoderma lucidum extract, 0.01 ml of mushroom mushroom extract, 0.05 ml of mushroom mushroom extract, 0.05 ml of shiitake mushroom extract and 0.01 ml of Cordyceps sinensis extract, Mushroom extract 0.05ml, and red ginseng extract 0.1ml, and ginseng extract 0.1ml of the anticancer fermentation method comprising a mushroom extract, characterized in that the mixed solution mixed in a volume ratio.

The method of claim 1,
The mushrooms, red ginseng and ginseng extracts are each of Ganoderma lucidum mushrooms, situation mushrooms, zinnia mushrooms, shiitake mushrooms, Cordyceps sinensis, tea mushrooms, red ginseng, and ginseng. Drying each of them separately placed in a container, and putting them in a hot air precursor chamber for drying at 50 ° C. for 12 hours;
The dried materials were separately extracted with a mixture of water and ethanol in a ratio of 1: 9, and extracted 1 to 2 times in a range of 65 to 105 ° C. with a reflux extractor, and filtered using a vacuum filter. , Anti-cancer fermentation method comprising a mushroom extract, characterized in that the filtrate was prepared through an extraction process of concentrating each 60-25 Brix using a vacuum concentrator.