KR20190066821A - A method of massively isolating and concentrating effective components from Grifola frodosa and a method of manufacturing powdered food for immune enhancement using the same - Google Patents

Abstract

The present invention relates to a method for massively isolating and concentrating active ingredients from Grifola frondosa and a production method of powdered food for immune enhancement using the same. Since the developed powdered food comprises active ingredients from the Grifola frondosa having an immune boosting effect and has improved immune boosting efficacy, health functional powdered food excellent for price compared to existing powdered food can be produced.

Description

Technical Field [0001] The present invention relates to a method for mass-isolating and concentrating a useful ingredient of a mushroom, and to a method for producing an immunoconjugate using the same,

The present invention relates to a method for mass-isolating and concentrating a useful ingredient of a mushroom, and a method for producing an immunity-enhancing wire using the same.

Grifola frondosa is a naturally occurring mushroom from late summer to autumn in the dead wood stump of broad-leaved trees such as water oak, chestnut, beech, etc., belonging to the genus Cryptospora, It is distributed in East Asia, Europe, North America and so on.

Fruiting body is composed of several branches in one stem and dozens of gums are attached to the tip of the stem to form a bundle like a pine cone. The mushroom is difficult to grow and the stem and the egg of fruity body develop gradually, .

Leaf mushroom has good taste and flavor and has been used as herbal medicine material since it has been known that it is excellent in blood pressure lowering, diuretic action, obesity treatment, tonic action, and antiemetic action in herbal medicine.

[0004] Since the commercial production of 325 tons of mushroom in Japan in 1981, its production has steadily increased to 1,500 tons in 1985, 8,000 tons in 1991, and 10,000 tons in 1993, It is produced more than 40,000 tons, and in Japan, it is a mushroom with high production and consumption next to top, top, and bottom.

For the bag cultivation technique using sawdust, there are research results such as appropriate light conditioning lifespan, medium composition study, and carbon dioxide concentration survey.

On the other hand, it is spreading worldwide due to various causative agents of common infectious diseases acquired by new strains and strains, and pathogens showing resistance to existing antibiotics are increasing. In general, the vaccine is effective against the infectious diseases. However, development of a natural immune enhancing material that increases resistance through strengthening of the immune system is required in consideration of ease of use and safety of the general public.

Sunsik was derived from the fact that the galleries of the Silla era used to mix and match seven grains as one of the convenient and nutritious natural health secrets to carry around while visiting the Mt. Seven kinds of grain refers to brown rice, glutinous rice, barley, black bean, black sesame, perilla, and yulmu. It is less burdened on the top, and it is easy to mix with fiber, oil and flavor. It has a characteristic that it has a mental health that is clear and has a gun. Since then, Sun-shik has developed into a high-protein nutritional formula prepared by priests and monks for a long period of time before starting the life of the capitals such as cave crucifers. Now, in addition to the traditional seven-grain cereal formula, the functional sour food is made by instantly powdering and selling dried vegetables, mushrooms, porridge, seafood, fruits, etc. .

Sunshine is a good food that has been used as food in the past and is often used as an emergency food for the baby. Thus, children and elderly people, such as people with a weak digestive system, such as regular food at all levels is good to eat, and digestive good even if the diet and special purpose supplementary nutrition helps.

As related prior art patents of the present invention, Korean Patent Laid-Open Publication No. 2010-0128170 discloses a technology relating to health supplements using leaf mushroom extract, The present invention relates to a technique for producing a solid fermented hot-water extract obtained by adding purified water to a fermented leaf mushroom and extracting it by hot water.

Korean Patent Laid-Open Publication No. 2010-0070777 discloses a method for mass production of cultured mycelia of leaf mushroom and immunopotentiating polysaccharide. The above technique is a method of cutting a freshly sectioned piece of mushroom fruiting body in a sterile zone and cultivating potato dextrose agar (PDA) After culturing the cells at 27 ° C for 4 weeks, the cells were separated from the PDA plate. Then, the cells were evenly pulverized using a sterilizing mill, and inoculated into a culture flask aseptically at 10% And a method for purifying a precipitate by ion exchange and gel filtration chromatography, respectively, by adding ethanol to a fruiting body, a culture filtrate, or a crude polysaccharide derived from a mycelium after culturing in an optimum medium.

The present invention has been made in view of the above-mentioned needs, and an object of the present invention is to provide a technique for massive isolation and concentration of useful components of mushrooms.

Another object of the present invention is to provide a method for producing an immunostimulatory line.

In order to accomplish the above object, the present invention provides a method for massively separating and concentrating useful components using fine pulverization and air classification.

In addition, the present invention provides a method of preparing a wire by applying a method of massively separating and concentrating useful components using fine pulverization and air classification of leaf mushroom to a wire making method.

The present invention also provides the above-mentioned compositions of the present invention.

In one embodiment of the present invention, the composition is preferably characterized by having an immunostimulating effect, but is not limited thereto.

Hereinafter, the present invention will be described.

Many patents related to processing have been filed for about 20 years, which is the history of domestics, and it is steadily increasing. However, most of the existing line products are at the level of selecting, mixing and selling materials based on traditional ingestion without sufficient research on food science and nutritional efficacy. Through the technique of the present invention, it is possible to meet the needs of consumers who demand health orientability rather than simple consumption.

In addition, the line product of the present invention can be developed as a high-grade line product that scientifically proved the physiological activity and the nutritional value of the line type manufactured through the raw materials proving its functionality and stability, unlike the conventional line type products.

In the conventional heat drying method, most of the original color, incense, etc. are lost during the drying process of the raw materials. Also, the drying energy efficiency is 25 ~ 30%, which increases the production cost, and the organic characteristics of the drying material are largely lost. On the other hand, far-infrared heating can transfer to the center of the material without destroying the constituents of the surface of the material, and it can prevent the loss of the organic properties longer than the drying using the ordinary heat source .

In the present invention, the wire fungus which has been finely pulverized into particles having a particle size of 150 to 200 mesh using a pulverizer and compared with the conventional wire, can be manufactured as a fine functional health line having excellent bioabsorbability as a fine powder.

As can be seen from the present invention, the development of the beneficial components such as immunity enhancement and the improved effect of the mushroom of Leaf mushroom can make the health functional line excellent in cost compared with the existing line type.

Figures 1 and 2 show the results of human growth factor array and human matrix metalloproteinase array analysis on the self-potential of cells through proteomics experiments.

Hereinafter, the present invention will be described in more detail by way of non-limiting examples. The following examples are intended to illustrate the present invention and the scope of the present invention is not to be construed as being limited by the following examples.

Example  1. The present invention Mushroom  Lumber growing

1) Preparation of wood

The oak tree used for cultivation of the mushroom was cut into 20 cm long oak tree with a diameter of about 10 to 15 cm. The cut oak tree was immersed in water for 24 hours, then placed in a heat-resistant PP bag, Lt; 0 > C for 180 minutes, then cooled sufficiently after 2 days, and inoculated with the seeds.

2) Inoculation of leaf mushroom seed

The liquid seed bacterium (30 g of soybean meal per 10 L of water, 300 g of sulfur sugar, 30 ml of vegetable oil) was inoculated with about 50 g per 100 g of the cut wood, and the saw-cut bacterium (V / V, 80% oak sawdust and 20% 100 ml is inoculated. If seeds are inoculated before March, they can be harvested in the current year. If they are inoculated after April, they will be harvested the following year.

3) Cultivation of mycelia of leaf mushroom

The hyphae of the mushroom are cultured for 90 days or more, preferably 90 to 100 hours, at a temperature of 22 ± 1 ° C. and a relative humidity of 65 to 75% under a dark condition while sufficiently ventilated.

4) Landfilling works

The seedlings cultivated with mycelia were inoculated into a container (length 50 cm, width 35 cm, height 30 cm) of soil in a plastic house shielded with 70% of the mycelium, and the soil was buried 5 cm thick After covering, cover the hardwood leaves again for moisturizing and give enough ventilation.

5) Cultivation management and harvesting

The soil where the logs are buried is irrigated so as not to dry, and harvested when the color of the mushroom fruiting body of the mushroom becomes yellowish brown and the size of the fruiting body maintains its maximum size. The fruiting body starts to occur when the temperature falls below 20 ° C on average, so the temperature should be kept below 20 ° C in order to produce fruiting bodies. Care should be taken to prevent the water from coming into direct contact with the fruiting body during its formation.

Example  2.

(1) Establishment of fine grinding and air classification conditions

We intend to utilize ultra fine grinding and air classification technology to massively isolate and concentrate the useful components contained in the leaf mushroom.

For this purpose, dried mushrooms were finely pulverized using a single track jet mill (STJ-100, Seishin, Japan) under feeding conditions of 1.2 to 2.0 kg / hr, pusher pressure of 0.72 MPa and grinding pressure of 0.70 MPa , And ultrafine pulverized mushroom powder were classified by air classification system (Spedic-250, Seishin, Japan) with different classifying wheel speed (ACWS, Air Classifying Wheel Speed) (2,000 ~ 6,500 rpm) . The fine powder recovered at the time of classification is indicated as "F (fine powder: Fine)" and the coarse powder having a much larger coarse particle size is denoted by C (coarse powder). For example, 6,500 F of powder recovered as fine powder at 6,500 rpm, and 6,500 C of unclassified powder. The classification conditions for each rpm are shown in Table 1 below.

Classification No. One 2 3 4 Throughput (kg / hr) 2.6 2.2 2.7 3.0 Classification rotor (rpm) 6,500 5,000 3,500 2,000 Bag flow meter differential pressure (mmAq) 300 300 270 300 Blower (Hz) 54.6 56.1 60.0 60.0 Operating time (min) 100 104 75 51

Table 1 shows the classification conditions for massive isolation and concentration of the mushroom.

(2) Particle size analysis and powder aptitude survey

The particle size of the fraction powder was analyzed by laser type particle size analyzer (LMS-30, Seishin, Japan).

(3) Establishment of standard and production standard of mushroom powder

Isolation and concentration of useful components of mushrooms When the optimum conditions are established, scale up to 5 kg, 20 kg and 100 kg, validate each process, and establish mass separation and concentration technology.

The nutrients and harmful components of the candidate material for the final prototypes separated and concentrated by the oil component are submitted to an authorized organization for analysis.

① Nine ingredients: calories, carbohydrates, sugars, protein, fat, saturated fat, trans fats, cholesterol, sodium

② Hazardous components

Microbiological tests: Common bacteria, Escherichia coli

Heavy metal test: lead, total arsenic, total mercury, cadmium

Analysis of residual pesticides: analysis of residual pesticide items corresponding to other agricultural products listed in Table 4 of "Food Standards and Standards"

(4) Analysis of the contents of useful components of concentrated mushroom fraction

- Leaf mushroom Indices: β-glucan

    Quantification of β-Glucan was measured using a 'Mixed-linkage beta-glucan kit' from Megazyme (The Bray Co.). Total glucan content was determined by adding 100 μL of 50% ethanol and 2 mL of 20 mM sodium phosphate buffer (pH 6.5) to 50 mg of the sample passed through a 0.5-mm sieve, stirring at 100 ° C. for 3 minutes, After cooling for 5 minutes, 100 μL of lichenase (5 U) was added and reacted by stirring at 50 ° C for 1 hour. After adding 2.5 mL of sodium acetate buffer (pH 4.0) and centrifuging (1,000 g, 10 min), 50 μL of the separated supernatant was mixed with the same amount of β-glucosidase (0.1 U) After adding 1.5 mL of GOPOD (glucose oxidase / peroxidase) reagent and reacting at 50 ° C for 20 minutes, the absorbance was measured at 510 nm, and the content of β-Glucan was calculated by the following equation.

Example 3. Leaf mushroom  Assessment of immunity enhancement efficacy by material: Using cell model In Vitro  Identification of efficacy and mechanism of action

In general, beta-glucan activates ancillary T-cell type 1 (Th1) to protect the body from viruses and bacterial infections. It also binds to receptors of human immune cells and changes its immune pattern to remove tumor cells or mutant cells, leading to immunological physiologically active substances.

The mushroom Beta Glucan has two types of immune functions. First, it directly activates the immune cells, macrophages, T cells, and NK cells. Second, it promotes the secretion of cytokines to support their function.

In addition, beta (1,3) D glucan is a natural substance and has no side effects. It has a slow degradation rate and functions in various ways for a long time in the body to activate immune cells. Like mushroom, yeast beta-glucan is also present in the cell wall, so it can not digest and absorb beta-glucan to eat yeast.

However, beta-glucan has immune-modulating activity as well as immune enhancement, so that it does not cause excessive immune response and little side effects are reported.

The market size will expand rapidly when it is used as a development material for patients using immunoglobulin using Beta Glucan, which is a useful ingredient of this mushroom.

In order to investigate the mechanism of the immune function of the cells, human growth factor arrays and human matrix metalloproteinase arrays are analyzed for the self-potential of cells through proteomics experiments.

Claims (5)

A method of massively separating and concentrating useful components using fine grinding and air classification of leaf mushrooms. A method for mass separation and concentration of useful components by using fine grinding and air classification of leaf mushroom to prepare a line preparation for immunity enhancement. 3. The method according to claim 2, wherein the wire has immunity enhancing effect. 4. A composition according to any one of claims 2 and 3, 5. The composition of claim 4, wherein the composition has an immunostimulating effect.

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