KR101851395B1 - Extract of graviola fermented by phellinus linteus and manufacturing method thereof - Google Patents

Abstract

The present invention relates to a Phellinus linteus fermentation graviola extract and a preparation method thereof and more particularly to a graviola extract obtained by inoculating and fermenting mushroom mycelia with a graviola plant as a substrate, Extract and a process for producing the same. According to the manufacturing method of the present invention, it is possible to maximize the content of the total polyphenol component and the total flavonoid component as the physiologically active substance contained in the graviola as well as the pharmacological effect of the mushroom, It is possible to utilize it as a high-value-added industry in foods, functional foods and medicines.

Description

Technical Field [0001] The present invention relates to an extract of graviola fermented with mushroom, and a method for producing the same. BACKGROUND OF THE INVENTION 1. Field of the Invention < RTI ID =

The present invention relates to a Phellinus linteus fermentation graviola extract and a preparation method thereof and more particularly to a graviola extract obtained by inoculating and fermenting mushroom mycelia with a graviola plant as a substrate, Extract and a process for producing the same.

Recently, consumers' perception of mushroom has expanded into not only edible but also medicinal and taste foods, and mushroom extracts and mycelium extracts and mycelial cultures have been found to be effective in the prevention and treatment of various diseases. It has emerged as an important biological resource for the development of functional foods and medicines. About 20,000 species of mushrooms are known worldwide, of which about 2,000 species can be developed for food. Approximately 992 species of mushrooms distributed in Korea are recorded. Among them, about 100 kinds of edible mushrooms and 50 kinds of mushrooms are found. Especially, more than 20 kinds of mushrooms having high toxicity are identified. More than 20 kinds of mushrooms can be cultivated in Korea, and have been proven to have anti-cancer, low cholesterol and low blood sugar.

The mushroom is also called woody mud mushroom. The mushroom has various shapes such as 6-12cm diameter, 2 ~ 10cm thick, semicircular shape, flat shape, round mountain shape, horseshoe shape. The dark brown hairs on the surface are short and densely packed, then disappear and grow to become crisp. In the early days of growth, mud clusters seem to be united, and after they are all grown, they are called tongue because they are out of their stumps in a tree stump. Since 1968, Kiekawa et al. Has been known to have anticancer effects, many studies have been carried out. In addition to the anticancer effects, various pharmacological effects such as immunomodulating effect, antioxidant effect, liver function improving action, hyperlipidemia improving effect, whitening effect are shown. For medicinal purposes, the moon is yellow or light yellow and is characterized by lack of flavor and aroma. Korea, Japan, Australia, and North America.

Graviola is known as an anticancer plant and contains highly active ingredients in leaves, stems, roots and seeds. Graviola is a small evergreen tree of about 5-6cm native to the Amazon and the tropics of South America and North America, including Brazil. The elliptical fruit of sweetness is prickly, and the skin of dark green or yellowish green has white flesh.

Graviola has been shown to reduce blood pressure, vasodilatation and cardioprotection through animal experiments. In particular, it is known that the active ingredients contained in leaves, seeds and stems of graviola are cytotoxic to cancer cells It is expected to be used in chemotherapy.

Graviola is ingested directly into tea, juice, capsule form, extract or raw food. However, because of the toxic alkaloid component contained in graviola, it may cause problems in blood pressure control and digestion during long-term use, and may cause abdominal pain and vomiting.

Therefore, it is required to develop a product that can maximize the physiological activity effect while facilitating ingestion of graviola by processing it in accordance with the taste of the consumer.

KR 10-2005-0003908 A KR 10-2015-0053357 A KR 10-2016-0001102 A KR 10-1581340 B

A problem to be solved by the present invention is to maximize the content of various physiologically active substances, especially polyphenols and flavonoids, contained in graviola by fermentation of Mycelia mushroom mycelium, The present invention provides a method for producing an extract of gravura fermented with mushroom fermentation and an extract of gravia fermented by the same, which can reduce an alkaloid component.

In order to solve the above-mentioned problems, the present invention relates to a method for producing a mushroom fermented graviola by inoculating and culturing the mycelium of mushroom mycelia into sterilized graviola to obtain a mushroom fermented graviola and then obtaining a mushroom fermented graviola extract from the mushroom fermentation graviola The present invention also provides a method for producing the graviola extract of the mushroom fermentation.

It is preferable that the graviola is selected from the group consisting of fruit, seed, stem, leaf and root of graviola.

The culture conditions are preferably 20 to 35 DEG C, 70 to 90% relative humidity, and 7 to 45 days for cultivation.

In addition, the present invention provides an extract of Ganoderma lucidum fermentation obtained by the above-described production method of the present invention.

According to the manufacturing method of the present invention, it is possible to maximize the content of the total polyphenol component and the total flavonoid component as the physiologically active substance contained in the graviola as well as the pharmacological effect of the mushroom, It is possible to utilize it as a high-value-added industry in foods, functional foods and medicines.

Fig. 1 is a flow chart showing the process for producing the mushroom fermented graviola extract of the present invention.

Hereinafter, the present invention will be described in detail.

The inventors of the present invention focused on various pharmacological activities of the mushroom to ferment the mushroom mycelium in a sterilized graviola plant to greatly increase the content of polyphenols and flavonoid components which are useful physiologically active ingredients contained in the graviola And that the content of the alkaloid component, which is a toxic substance contained in the viola, can be reduced, thereby completing the present invention.

Accordingly, the present invention relates to a method for producing a mushroom fermented graviola extract, which comprises obtaining a mushroom fermented graviola by inoculating and culturing the mycelium of mushroom mycelia into sterilized graviola, and then extracting the mushroom fermented graviola extract from the mushroom fermentation graviola And a manufacturing method thereof.

Preferably, the condition mushroom of the present invention is Phellinus linteus , which is known as woody mud mushroom.

The mushroom mycelium of the present invention can be produced by seed culture of the mushroom seedlings.

As a preferred example, the mushroom seedlings are inoculated into a PDA (Potato Dextrose Agar) medium, and then cultured in a subculture at about 4 weeks intervals at a temperature of about 25 ° C. .

The seed culture is a step of culturing the mycelia mycelium cultured in storage prior to the main culture. In a preferred embodiment, the seed culture can be obtained as a liquid culture by inoculating seed culture, i.e., mycelium, into a culture medium for mushroom mycelium, and then culturing the culture in a rotary shake.

The graviola may be selected from the group consisting of fruit, seed, stem, leaf and root, and more preferably selected from fruit, stem or leaf, although graviola can utilize whole graviola plants. And most preferably leaves.

The graviola can be sterilized by washing the prepared graviola, dehydrating and drying it, and applying various known sterilization methods. In a preferred embodiment, the sterilization can be autoclaved at 121 DEG C for 30 minutes.

The method of inoculating the mycelium of the present invention to the sterilized graviola is aseptically inoculating a predetermined amount of the liquid culture obtained by the above described mushroom culture to the sterilized graviola.

Preferably, the culturing is carried out at a temperature of 20 to 35 DEG C, a relative humidity of 70 to 90%, and a culturing period of 7 to 45 days. As a preferable example, For 30 days at a temperature of 80% relative humidity.

The mushroom fermentation graviola is a form in which the surface of the graviola plant is entirely covered with the mycelium of mycelia, from which the graviola fermented graviola extract is prepared.

The solvent used for the extraction may be water or lower ethanol, but is preferably hot water extraction. As a preferable example, the supernatant can be taken by adding hot water of about 5 to 150 times volume to the obtained mushroom fermentation graviola, shaking for 1 to 5 hours, and centrifuging. The above process may be performed 1 to 5 times, preferably 3 times.

The gravy fermented graviola extract of the present invention can be prepared through the above process.

Accordingly, the present invention provides a mugwort fermented graviola extract obtained by the above production method of the present invention.

The gravure fermented graviola extract prepared according to the method of the present invention contains various components exhibiting various pharmacological effects, which are present in a large amount in the mycelium of mushroom by the fermentation of the mushroom, and the total amount of the phyto- Phenol, and total flavonoid, and the alkaloid component content, which is a toxic substance, is greatly reduced.

The above-mentioned mushroom fermented graviola extract may be prepared in liquid or powder form suitable for the purpose of use, and in the case of powdery form, the mushroom fermented graviola extract may be pulverized by concentration under reduced pressure.

The present invention relates to a method for the treatment and prophylaxis of various types of malignant tumors such as colon cancer, breast cancer, prostate cancer, liver cancer, pancreatic cancer or stomach cancer, bacterial diseases, Diabetes, diarrhea, dysentery, edema, neuralgia, neurotoxicity, rheumatoid arthritis, aging, various inflammatory diseases, insomnia, diabetes mellitus, hypertension, depression, , Asthma, and the like.

The pharmaceutical composition of the present invention may be formulated into oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, injections or creams of sterilized injection solutions, lotions, etc. And external preparations for skin, and may be administered by various routes including oral administration or intravenous, intraperitoneal, subcutaneous, rectal, topical or skin application. Such pharmaceutical compositions may further comprise carriers, excipients or diluents, and examples of suitable carriers, excipients or diluents that may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, But are not limited to, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, amorphous cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, And the like. In addition, the pharmaceutical composition of the present invention may further include a filler, an anti-coagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent, an antiseptic, and the like.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will depend on the type of disease, severity, The sensitivity to the drug, the time of administration, the route of administration and the rate of release, the duration of the treatment, factors including co-administered drugs, and other factors well known in the medical arts. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without side effects, which can be easily determined by those skilled in the art.

In a preferred embodiment, the effective amount of the active ingredient of the pharmaceutical composition of the present invention may vary depending on the age, sex, and body weight of the patient, and is generally 1 to 5,000 mg, preferably 100 to 3,000 mg per day, It can be administered every other day or one to three times a day. However, the dosage may not be limited in any way because it may be increased or decreased depending on route of administration, severity of disease, sex, weight, age, and the like. The pharmaceutical composition of the present invention can be administered to a subject through various routes. All modes of administration may be expected, for example, by injection into the skin, oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections. &Quot; Administration "in the present invention means providing the patient with the desired material in any suitable manner, and the route of administration of the pharmaceutical composition of the present invention may be applied through any general route as long as it can reach the target tissue, Or parenterally. The composition of the present invention may also be administered using any device capable of delivering an effective ingredient to a target cell. In the present invention, the term "object" includes, but is not limited to, human, monkey, cow, horse, sheep, pig, chicken, turkey, quail, cat, dog, mouse, rat, rabbit or guinea pig , Preferably a mammal, more preferably a human.

The present invention also relates to a method for the treatment of various types of malignant tumors such as colon cancer, breast cancer, prostate cancer, liver cancer, pancreatic cancer or stomach cancer, bacterial The present invention relates to a pharmaceutical composition for preventing or treating inflammatory diseases such as inflammatory diseases, fungal diseases, hypertension, depression, convulsions, viral diseases, pain, gastrointestinal disorders, heart failure, chest pain, cold, diabetes, diarrhea, , Insomnia, asthma, and the like.

The health functional food of the present invention can be variously used for foods and beverages effective for prevention or improvement of the diseases. Examples of foods containing the active ingredient of the present invention include various foods, beverages, gums, tea, vitamin complex, health supplement foods and the like, and they can be used in the form of powder, granule, tablet, capsule or beverage . The active ingredient of the present invention may generally be added in an amount of 0.01 to 15% by weight of the total food, and the health beverage composition may be added in a proportion of 0.02 to 10 g, preferably 0.3 to 1 g, based on 100 ml.

The health functional food of the present invention may contain, as an essential ingredient, the above-mentioned active ingredient in a prescribed ratio, as well as a food-acceptable food supplementary additive such as natural carbohydrate and various flavoring agents as an additional ingredient. Examples of the natural carbohydrate include sugar sugars such as glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of the flavor agent include tau martin, rebaudioside A, glycyrrhizin, saccharin, and aspartame. The proportion of the above-mentioned flavoring agent is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the health functional food of the present invention.

In addition to the above, the health functional food of the present invention may contain various kinds of nutrients, vitamins, minerals, flavors such as synthetic flavors and natural flavors, colorants and heavy stabilizers, pectic acid and its salts, alginic acid and its salts, Thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the health functional food of the present invention may contain flesh for producing natural fruit juice, fruit juice drink, vegetable drink and the like. These components may be used independently or in combination. The proportion of such additives is generally selected in the range of 0.01 to about 20 parts by weight per 100 parts by weight of the active ingredient of the present invention.

Hereinafter, the present invention will be described in more detail with reference to specific examples. The following examples illustrate one preferred embodiment of the present invention, and it is apparent that the scope of the present invention is not limited by the following examples.

[Example]

Example 1. Preparation of Graviola Extract from Fermented Mushroom Fermented

Phellinus linteus was cultivated in a PDA (Potato Dextrose Agar, Difco, Co., USA) medium at 25 ° C. using a strain in a farm, Bioscience Inc., and subcultured at intervals of 4 weeks. 4 < [deg.] ≫ C. In a 500 ml flask, 100 ml of mushroom mycelial solid medium (1 cm x 1 cm, 3 pieces) was inoculated with submerged mycelia of mycelium, and then transferred to a 7 ml flask in a rotary shaking incubator (Daihan Labtech Co.Korea, 25 ± 1 ° C, 130 rpm) Day, and used as liquid seeds.

Graviola was purchased from Daegu herb medicine market, and graviola leaves were washed, washed with water to remove impurities and dehydrated. 100g of graviola leaves were placed in a 3L culture container, sterilized at 121 ° C for 30 minutes, And 60 ml of the liquid seed thus prepared was aseptically inoculated into 100 g of sterilized graviola. Fermentation was carried out for 30 days in an incubator at 25 ° C and 80% relative humidity. After fermentation, 10 times volume of hot water was added, and the mixture was shaken for 2 hours and then immersed for 72 hours. After centrifugation at 10,000 rpm for 10 minutes, the supernatant was taken. After repeating this three times, the extract was concentrated under reduced pressure to prepare a powder Respectively.

Example 2. Analysis of components of Graviola extract fermented with mushroom mushroom

Table 1 shows the compositional analysis results of the grape viola extract obtained in Example 1 and the graviola leaf extract not fermented with the mushroom. The total polyphenol content was determined by adding 50 μl of Folin-ciocalteau and 100 μl of a saturated solution of Na ₂ CO _{3 to} 400 μl of the extract solution, and incubating the solution at room temperature for 1 hour and measuring the absorbance at 725 nm. Tannic acid was used as a standard reagent. The total flavonoid content of each sample was measured by stirring for 18 hours in methanol, 4 ml of 90% diethylene glycol was added to 400 μl of the filtered extract, 40 μl of 1N NaOH was added, and the absorbance at 420 nm was measured at 37 ° C. for 1 hour. As a standard reagent, rutin was used. Reducing sugar was determined by DNS method and total sugar was quantified by phenol-sulfuric acid method.

Analysis of Components of Graviola Extract from Fermented Mushroom (Unit: mg / g) division Content (mg / ml) Total polyphenol Total flavonoid Per capita Reducing sugar Mushroom fermentation
Graviola extract 250 ± 21.5 42 ± 4.05 320 ± 15.0 67 ± 0.43 Graviola leaf extract 155 ± 5.15 18 ± 2.52 352 ± 11.2 58 ± 6.55

As shown in Table 1, the total polyphenol content of the graviola extract fermented with the mushroom was 250 ± 21.5 mg / g, while that of the graviola leaf extract, which was not fermented with the mushroom, was 155 ± 5.15mg / g, and the total flavonoid content was about 2 times higher than that of the mushroom fermented graviola extract. From these results, it can be confirmed that a large amount of total polyphenols and total flavonoids are produced as useful phytochemical components when the graviola leaves are fermented with mushroom.

Example 3. Analysis of Alkaloid Components of Graviola Extract from Fermented Mushroom Fermentation

The graviola leaf extract obtained from the above Example 1 and the non-fermented graviola leaf extract as a control were used as a control group (see Jang Seung-yeop, Ryu Kyung-soo, To determine the alkaloid content.

Alkaloid content (%) of graviola fermented with mushroom division Total alkaloid content (%) Situ mushroom fermented graviola extract 0.76 ± 0.1 Graviola leaf extract 1.53 ± 0.2

 As shown in Table 2, it was confirmed that the content of alkaloid known as a toxic substance was reduced by about 50% as compared with that of non-fermented graviola leaves in the mushroom fermented graviola extract. It is expected that the alkaloid component, which is a nitrogen compound, is converted into another metabolite during the fermentation process.

Claims (4)

The mushroom mycelium was inoculated into sterilized graviola and cultured at a temperature of 20 to 35 ° C, a relative humidity of 70 to 90%, and a cultivation period of 7 to 45 days to obtain a mushroom fermentation graviola. The mushroom fermentation graviola A method for reducing an alkaloid component contained in graviola, which is obtained by fermenting graviola extract. The method according to claim 1, wherein the graviola is selected from the group consisting of fruit, seed, stem, leaves and roots of graviola. delete delete