KR101755338B1 - Process for preparation of grifola frondosa fruit body with twice fermentation and functional grifola frondosa fruit body fermentative products - Google Patents

Abstract

In order to improve the bioavailability and anticancer immune activity of leaf mushroom fruiting bodies, the present invention relates to a method for producing a fungus body, comprising the steps of inoculating a water extract obtained after primary solid culture with edible fungus into a dried mushroom fungus, And to a method for producing the fermented extract of Mushroom mushroom having improved anti-cancer immunity activity.
The fermented product of leaf mushroom fruiting body according to the present invention is improved in bioavailability and anticancer immune activity by the combination of the first solid fermentation and the second liquid fermentation such as the fermentation of the second liquid to produce various materials such as general foods, And it can be used as a functional cosmetic material because the molecular weight of the useful substance is reduced because it is useful for maintaining freshness of fermented food and fortifying nutrition.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a method for producing a fermented product of a two-stage fermented leaf mushroom fruiting body and a fermented product of the anticancer functional fucoidan mushroom fruiting body produced thereby,

The present invention relates to a method for biodegrading and transforming leaf mushroom fruiting bodies through a two-stage fermentation process, and to a mushroom fruiting body having improved anticancer and immunological function.

Grifola frondosa is a taxonomic species belonging to Aphyllphorales , Polyporaceae , and Grifola . It is a mushroom belonging to the genus Grifola . It grows around the branches and roots of large broad-leaved trees during summer and autumn. It is the white part of the wood which occurs as the mushroom. Fruiting bodies are composed of a large number of goblets divided into several small bases, which are formed from one large basin. There are also large mushrooms of which the total diameter of the gut is more than 30 cm and the weight is more than 3 kg. The surface is dark brown to gray and has a radial fiber pattern.

The D-fraction purified from the polysaccharide of the mushroom has been commercialized in the United States and Japan as β-1, 3-glucan having 6 types of beta 1, 6, and antihypertensive, anti- And hyperlipidemia have been reported. As a result of administration of β-glucan extracted from mycelium of mushroom to mice transplanted with cancer cells such as fibrosarcoma, L1210 leukemia, and P388 leukemia, natural killer cells and macrophages ) To increase the activity of inhibiting cancer. Based on these studies, the polysaccharide of the leaf mushroom suppressed the cancer cells and obtained the US FDA approval in 1998 as an anti-cancer adjuvant.

However, most of the conventional techniques have been used by using hot water, organic solvent extraction, or powdering of leaf mushroom fruiting bodies. In this case, the bioavailability of the leaf mushroom useful materials is lowered and the effect of enhancing the anti-cancer immunity activity is reduced.

The object of the present invention is to improve the bioavailability and anticancer immune activity of leaf mushroom fruiting body by adding Aspergillus fungi, an edible fungus, The present invention relates to a method for producing a fermented extract of Leucocephalica mushroom having improved anti-cancer immunity by extracting, concentrating and pulverizing a liquid culture obtained by inoculating a water extract obtained from a primary solid culture with an oryzae will be.

One aspect of the present invention relates to a method for producing a fermented product of two-stage fermented mushroom fruiting body comprising the following steps.

1) preparing a dried powder of mushroom fruiting body; And

2) To the dried powder of the above mushroom fruiting body, Aspergillus oryzae ) to obtain a primary fermented product;

3) an extraction step of adding water to the primary fermented mushroom and sterilizing it at a high temperature to obtain a mushroom water extract;

4) a secondary fermentation step in which Leuconostoc mesenteroides or a lactic acid bacterium and a useful microorganism complex (EM) are inoculated to the above-mentioned leaf mushroom water extract to obtain a biomarker of leaf mushroom; And

5) filtration and concentration of the leaf mushroom bioconverting material followed by drying to obtain a solid fermented mushroom;

In one example, the dried powder of the mushroom of the leaf mushroom can be obtained by drying the mushroom fruiting body of the mushroom of the leaf mushroom at a temperature of 30 to 40 ° C. and drying within 10% of the moisture content.

In one example, the yeast mold ( Aspergillus oryzae ) may be cultured in a PDB medium at 30 DEG C and 120 rpm for 7 days with shaking.

In one example, the primary fermentation step may be performed by incubating for 7 days at 25 < 0 > C.

In one example, the extraction step may be performed by adding 3 to 5 times the weight of water to the primary fermented mushroom, sterilization at 120 DEG C for 15 minutes, and cooling at room temperature.

In one example, the secondary fermentation step to obtain the biomass conversion product of the leaf mushroom is performed by culturing the mixed microorganism strain of Leuconostoc mesenteroides or lactic acid bacteria and the useful microorganism in a MRS liquid medium and a Manganese mixed medium at 37 DEG C for 1 day 2% by weight of the raw material, and culturing at 30 to 37 ° C for 2 to 3 days.

In one example, the materialization step may include the following steps.

5-1) Extracting the secondary fermented mushroom by hot water extraction at 95-100 ° C for 30 minutes to 1 hour and filtering;

5-2) concentrating the supernatant obtained by centrifuging the filtrate to a concentration of 20-40 Brix under reduced pressure to obtain a concentrate;

In one example, after the concentration step, the selected drying step may be further performed.

5-3) drying the concentrate by lyophilization or spray-drying; Or the concentrate is added with 5 to 10 times by weight of ethanol and the mixture is allowed to stand for 24 hours at 4 째 C, followed by centrifuging the resulting precipitate, followed by lyophilization;

Another aspect of the present invention relates to a fermented product of leaf mushroom fruiting body produced by the above method.

Further, one aspect of the present invention relates to an anticancer composition comprising the fermented product of the mushroom fruiting body. The anticancer composition may be a solid cancer or a composition for multiple cancers, but is not limited thereto.

The fermentation of the two-stage fermented leaf mushroom fruiting body according to the present invention can be carried out by biotransformation of the leaf mushroom through two stages of fermentation using an edible fungus and lactic acid bacteria, Activity. Therefore, it can be applied to various materials such as general food related to immunity enhancement, health functional foods, medicines and the like.

In addition, the fermented food has a high utilization value for the purpose of maintaining freshness and nutrition, and the molecular weight of the useful substance is reduced, so that it can be utilized as a functional cosmetic material.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a flow chart of a process for manufacturing a fermented product of a foliar mushroom fruiting body according to an embodiment of the present invention.

Unless defined otherwise, all technical terms used in the present invention have the following definitions and are consistent with the meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Also, preferred methods or samples are described in this specification, but similar or equivalent ones are also included in the scope of the present invention. The contents of all publications referred to herein are incorporated herein by reference.

The term "about" is used herein to refer to a reference quantity, a level, a value, a number, a frequency, a percent, a dimension, a size, a quantity, a weight, or a length of 30, 25, 20, 25, 10, 9, 8, 7, Level, value, number, frequency, percent, dimension, size, quantity, weight or length of a variable, such as 4, 3, 2 or 1%.

Throughout this specification, the words "comprises" and "comprising ", unless the context requires otherwise, include the steps or components, or groups of steps or elements, Steps, or groups of elements are not excluded.

In the present specification, "Grifolafrondosa" is a taxonomic species belonging to the family Aphyllphorales, Polyporaceae and Grifola in summer and autumn, It is a white rot fungus which is distributed in Korea, Japan, Europe, America and so on.

Hereinafter, the present invention will be described in detail with reference to Fig.

A method for producing a fermented product of a two-stage fermented mushroom fruiting body according to an embodiment of the present invention includes the following steps.

S1) a preparation step of preparing a dry powder of fruiting body of leaf mushroom; And

S2) The dried mushroom powder of Aspergillus oryzae ) to obtain a primary fermented product;

S3) an extraction step of adding water to the fermented leaf mushroom and sterilizing it at a high temperature to obtain a leaf mushroom water extract;

S4) a second fermentation step in which Leuconostoc mesenteroides or a lactic acid bacterium and a useful microorganism-based strain (EM) are inoculated in the water extract of Leaf mushroom to obtain a leaf mushroom bioconversion product; And

S5) filtration and concentration of the biotransformation product of the leaf mushroom, followed by drying to obtain a solid fermented product;

In addition, the step S5 of forming the mushroom comprises the following steps.

(S5-1) extracting the secondary fermented mushroom with hot water at 95 to 100 ° C for 30 minutes to 1 hour, and then filtering;

(S5-2) concentrating the supernatant obtained by centrifuging the filtrate to 20-40 Brix under reduced pressure to obtain a concentrate; And

(S5-3) drying the concentrate by lyophilization, spray drying, or 5 to 10 times by weight of an organic solvent, followed by cooling at 4 ° C for 24 hours, centrifuging the precipitate, and lyophilizing the precipitate;

The leaf mushroom is a fragrant edible mushroom. Especially in Japan, when it finds a leaf mushroom, it is called a mushroom which is called dancing mushroom. , B2, C, and D are known, and it is known that sterols such as ergosterol and β-glucan which is excellent in promoting immunity activity are contained in large amounts. In general, β-glucan in other mushrooms has a chemical structure of β-1, 4 bonds in D-glucans, whereas β-1, 6 bonds in β-1, 3 bonds in leaf mushrooms.

The organic solvent usable in the present invention may be selected from ethanol, methanol, butanol, ether, ethyl acetate, chloroform or a mixed solvent of these organic solvents and water. In view of the safety of raw materials, Of ethanol is used. After obtaining the extract by using the solvent, a liquid substance can be obtained by freezing, heating and filtering at room temperature by a conventional method known in the art. Alternatively, the solvent can be evaporated, spray dried or lyophilized But the present invention is not limited thereto, and any method generally used in the art can be used without limitation.

Another aspect of the present invention is a fermented product of leaf mushroom fruiting body produced by the above method and a composition comprising the same.

In a composition according to an aspect of the present invention, the composition may be an anticancer agent containing a fermented product of a leaf mushroom fruiting body as an active ingredient.

Particularly, when the fermented product of the mushroom fruiting body according to the present invention is administered to the human body, there is no fear of side effects as compared with other synthetic drugs because it is a natural extract. When the fermented mushroom fruiting body of the present invention is orally administered to rats, As a result, it was proved to be a safe material and its stability was secured.

In addition, the present invention provides a health food containing as an active ingredient a fermented product of leaf mushroom fruiting body.

The health food is used in combination with other food or food additives other than leaf mushroom, and can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to its use purpose, for example, prevention, health or therapeutic treatment.

The effective dose of the fermented product of the mushroom fruiting body contained in the above health food can be used in accordance with the effective dose of the above pharmaceutical composition. However, for the purpose of health and hygiene, or for a long term intake for health control purposes, And it is clear that the active ingredient can be used in an amount exceeding the above range since there is no problem in terms of safety.

There is no particular limitation on the type of the health food, and examples thereof include meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, Drinks, alcoholic beverages and vitamin complexes.

In one aspect of the present invention, the composition may comprise from 0.001 to 10% by weight of fermented mushroom fruiting body based on the total weight of the composition.

If the content of the extract is less than 0.001% by weight based on the total weight of the composition, the anticancer, antibacterial and anti-inflammatory effects are insignificant. If the content exceeds 10% by weight, no marked increase in the effect is observed. In view of the above, the composition of the present invention is used in an amount of 0.005 to 9.5% by weight, 0.01 to 9% by weight, 0.03 to 8.5% by weight, 0.05 to 8% by weight, 0.07 to 7.5% by weight, 7 to 10 wt%, 0.1 to 6.5 wt%, 0.3 to 6 wt%, 0.5 to 5.5 wt%, or 0.7 to 5 wt% of the extract.

Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited to these embodiments.

[ Example  One]

1-1. Mushroom  Drying and crushing of fruit bodies

Dried mushroom foliage is dried at 30 ~ 40 ℃ with moisture content within 10% and cut into 3x4 cm size.

1-2. Mushroom  Cultivation of fruiting body

Steam sterilized dried mushrooms at 100 ~ 121 ℃ for 15 ~ 30 minutes, and then 10% by weight of Aspergillus oryzae cultured in PDB medium at 30 ℃ and 120 rpm for 7 days with shaking was added to the mushroom. The solid medium of Aspergillus oryzae inoculated with the mushroom was incubated at 25 ° C for 7 days.

1-3. Mushroom From the solid fermentation product  Water extract manufacturing

3 to 5 times the weight of water is added to the solid fermented mushroom obtained in 1-2, and the mold is sterilized at 120 ° C for 15 minutes and then cooled at room temperature.

1-4. Mushroom  solid Fermented  Secondary liquid fermentation

The solid fermentation water extract prepared in 1-3 is used to inoculate Lecuconostoc mensentroides or a lactic acid bacteria complex (EM bacterium) to produce a biomass converted mushroom. Leuconostoc mensentroides and complex strains are incubated at 37 ℃ for 1 day in MRS liquid medium and Manganese medium, respectively. After 2% by weight of the raw materials, they are incubated at 30 ~ 37 ℃ for 2 ~ 3 days.

1-5. Mushroom Fermented  Materialization

The fermented mushroom prepared in 1-4 is subjected to hot extraction at 95-100 ° C for 30 minutes to 1 hour and then filtered. The filtrate is centrifuged and the supernatant is concentrated under reduced pressure to 20-40 Brix. This concentrate is lyophilized or spray-dried to obtain a material. Alternatively, 5 to 10 times by weight of ethanol is added to the concentrate and the mixture is allowed to stand for 24 hours at 4 ° C. The resulting precipitate is centrifuged and the precipitated material is lyophilized.

<Experimental Method>

1. Sample

The fermented mushroom fermented product used in the present experiment was a lyophilized sample of the ethanol precipitate prepared in Example 1.

2. Experimental animals

20 ± 2 g of 3 weeks old male ICR mice were used for experiments for survival, tumor suppression and immune cell proliferative capacity after stabilization for 1 week. Ten mice or more were used as the experimental group in the combat experiment in which the cancer cells were transplanted after the sample administration. Feeds (for mice) and water were allowed to eat freely during the experiment and the room temperature was maintained at 24 ± 2 ℃.

3. Culture of tumor cells

The sarcoma-180 tumor cells were aseptically harvested, washed with 4% Hank's balanced salt solution (HBSS), and subcultured in ICR mice at a concentration of 1 x 106 cells / ml. The subcultured sarcoma-180 was collected from the abdominal cavity with pasteur pipettes, placed in a 50 ml conical tube, centrifuged at 1200 rpm for 10 min, washed three times with HBSS buffer, and used for the development of multiple cancers and solid tumors.

4. On multiple cancers  Anti-cancer test

10 weeks old ICR mice of average weight of 25.1 ± 0.2g were stabilized in a culture room of 24 ± 2 ℃ for 1 week and the fermented mushroom samples were dissolved in physiological saline and 200kg / Thereafter, 3 × 10 ⁵ cells / ml of sarcoma-180 was transplanted into the abdominal cavity. After the transplantation, 150 ~ 200mg / kg of the sample was administered by the same method, and the mean survival days (MST) and survival days elongation (ILS) were determined 30 days after transplantation using the following equation .

(ILS%) = (average survival days in the subject-administered group / average survival days in the non-administered group) x 100

5. In solid rock  Anti-cancer test

10 male ICR mice of 3 to 4 weeks of age, weighing 24.8 ± 0.2 g, were pretreated with the same conditions as those of multiple cancers. Then 2.5 × 10 ⁶ cells / ml of sarcoma-180 was injected subcutaneously in the right inguinal region of the ICR mice Respectively. After transplantation, the samples were administered in the same manner as the multiple cancers. After 30 days, the solid tumors were removed and the weight was measured. The rate of tumor growth inhibition was determined by the following formula, which was used as an index of the anti-cancer effect.

Tumor growth inhibition rate (IR,%) = (1-average tumor weight in the subject administration group / average tumor weight in the non-administration group) x 100

6. Cytotoxicity experiment

(Human cell line derived from human lung cancer), SKOV-3 (human ovarian cancer derived cell line), SKMEL-2 (human colorectal cancer derived cell line), HCT 15 (human colon cancer derived cell line) , And XF 498 (human central nervous system cancer cell line). These cell lines were separated from the adhered surface with trypsin-EDTA solution for the purpose of the experiment and were dispensed in a 96-well flat-bottom microplate (Falcon) such that the number of cells per well was 4-5 × 10 4 cells / ml. The cells were incubated in a CO2 incubator for 24 hours and adhered to the bottom. The medium (RPMI medium) was removed with an aspirator and the medium was incubated at 6 concentrations (6.25, 12.5, 25, 50, Ml) was added to the wells containing the cancer cells in triplicate (100 μl each), and the cells were further cultured for 46 hours. Then, the cells were added with the drug according to the sulforhodamine-B (SRB) assy method (T) and the number of cells (C) when cultured for 48 hours with a drug-free medium and the number of cells (T) cultured for 48 hours with the drug at each concentration were measured. Cell proliferation rate was calculated by the following formula.

Cell proliferation rate (%) = ((T - Tz) / (C - Tz)) x 100

7. In vitro Lymphocyte  Cell proliferation experiment

Cell viability was investigated by measuring absorbance at 450 nm without the use of radioactive isotopes using the principle that tetrazolium salt MTT reagent can form orange formazan only in living cells.

A single cell culture was prepared from a spleen aseptically collected from untreated mice using RPMI 1640 solution containing 10% fetal calf serum (FCS: GIBCO), 2 mM L-glutamine, 15 mM HEPES and 5 g / ml gentamycine The following splenocyte culture medium was adjusted to a concentration of 2 × 10 ⁵ cells / ml, and 100 μl of the mixture was added to a 96-well microtiter plate. MTT labeling reagent was added with 50 ㎕ of MTT labeling mixture (final concentration: 0.3 ㎍ / ㎖) containing electron coupling reagent, and the cell proliferation was examined by measuring the absorbance at 450 nm by pretreating the cells.

8. Macrophage spreading ability  Measure

Peritoneal macrophages were adjusted to 1 × 10 ⁵ cells / well and mixed with the samples in a 96-well plate for 48 hours at 37 ° C. The viability of the macrophages was observed. The activity was determined as a percentage of 300 macrophages Respectively.

[ Experimental Example  One] Mushroom Fermented On multiple cancers  Anticancer effect

As shown in Table 1, the mean survival days (MST) of the control group were significantly higher than those of the control group, as shown in Table 1, after implantation of 3x10 ⁵ cells / ml sarcoma-180 tumor cells into the ICR mice after the one- ) Were 19.5 days, and the survival days were prolonged by 131% and 162%, respectively. The MST was prolonged by 6 days and 12 days, respectively, and the 150mg / kg and 250mg / kg groups of the mushroom fermented product were 25.5 and 31.5 days respectively. The number of viable mice survived until 30 days after the fermentation. These results indicate that the antifungal activity of the mushroom fermented rats is comparatively excellent.

Anticancer effect of sarcoma-180 fermented mushroom on multiple cancers Treatment Dose
(Mg / kg) Route MST (day)
(Average survival days) ILS (%)
(Survival days elongation rate) 30 days survival Control group saline p.o. 19.5 - 0/10 Fermented mushroom 150 p.o. 25.5 131 ↑ 5/10 Fermented mushroom 250 p.o 31.5 162 ↑ 6/10

[ Experimental Example  2] Mushroom Fermented In solid rock  Anticancer effect

As shown in Table 2, the mean tumor weight of the control group was 1.68 ± 0.31 g, and the fermented product of the mushroom fermented product was 150 mg / kg, The dose of 250mg / kg was 0.68 ± 0.25g and 0.45 ± 0.12g, respectively, and tumor inhibition rates were as high as 65.1% and 74.8%, respectively.

Anticancer Activity against Sarcoma-180 Solid Carcinoma of Fermented Mushroom Treatment Dose
(Mg / kg) Route Tumor weight
(Tumor weight, g) Inhibition rate
(Inhibition rate,%) Control group saline p.o. 1.68 ± 0.31 - Fermented mushroom 150 p.o. 0.68 ± 0.25 65.1 Fermented mushroom 250 p.o 0.45 ± 0.12 74.8

[ Experimental Example  3] Mushroom Fermented  Cytotoxicity test results

SKOV-3, SKMEL-w, HCT 15, and XF 498 at concentrations of 6.25 ㎍ / ㎖ to 200 ㎍ / ㎖ in the SRB assay for the hot-water extract of the mushroom fermented product. As shown, the mushroom fermented product showed no direct cytotoxicity against all cancer cells. These results indicate that long-term ingestion as a food ingredient has little risk of side effects.

Cytotoxicity of fermented mushroom on various cancer cell lines sample
density
(占 퐂 / ml) Proliferation rate of various cancer cells
A549
(Lung cancer cell line) SK-OV-3
(Ovarian cancer cells) SK-MEL-2
(Colorectal cancer) XF498
(Central nervous system cancer) HCT15
(Colon cells) 6.25 101.3 101.7 103.5 102.7 106.3 12.5 101.1 101.6 103.2 102.5 105.4 25.0 100.9 101.4 102.4 102.1 103.2 50.0 100.8 100.8 102.1 101.8 101.4 100.0 100.3 100.1 100.6 101.2 100.8 200.0 99.8 99.7 99.8 99.9 100.4

[ Experimental Example  4] Mushroom Fermented  Macrophage activity

The activity of macrophages according to the MTT method was measured at 50, 100 and 200 μg / ml concentrations of the fermented mushroom fermented product. The results are shown in Table 4. The concentration of proliferative acti vity and spreading activity of macrophages increased in a dose dependent manner.

Effect of macrophage activity on fermented mushroom Treatment (/ / ml) ^a Proliferative activity ^b Spreading ability ^c Control 1.00 + - 0.13 ^d 3.71 ± 0.25 ^* 1.00 ± 0.12 ^d Standard LPS (0.5) 3.71 ± 0.25 ^* 5.04 ± 0.28 ^* Fermented mushroom 50 2.41 ± 0.23 2.81 ± 0.25 100 2.83 ± 0.22 3.35 ± 0.45 200 3.41 + - 0.41 3.90 + 0.18

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (11)

A preparation step of preparing a dried powder of fried mushroom fruiting body; And
The above dried mushroom fruiting body powder was inoculated with a culture solution of Aspergillus oryzae cultured in a PDB medium at 30 ° C and 120 rpm for 7 days and cultured at 25 ° C for 7 days to obtain a primary fermented product A fermentation step;
An extraction step of adding 3 to 5 times by weight of water to the primary fermented product of the leaf mushroom, sterilization at 120 DEG C for 15 minutes and cooling at room temperature to obtain a water extract of Mushroom mushroom;
The Leuconostoc mesenteroides or the lactic acid bacteria and the useful microorganism complex (EM) which were cultured in the MRS liquid medium and the Manganese mixed medium at 37 ° C for 1 day were inoculated 2% Followed by incubation at 30 to 37 ° C for 2 to 3 days to obtain a biomass conversion product of leaf mushroom; And
A method for producing a fermented product of a two-stage fermented green mushroom fruiting body comprising the step of filtering and concentrating the leaf mushroom bioconverting material, followed by drying to obtain a solid fermented product.
2. The method according to claim 1,
Wherein the dried powder of the mushroom fruiting body is obtained by drying the mushroom fruiting body of leaf mushroom at a temperature of 30 to 40 캜 at a moisture content of less than 10% and then cutting it. delete delete delete delete The method according to claim 1, wherein, in the materializing step,
Extracting the secondary fermented mushroom with hot water at 95 to 100 ° C for 30 minutes to 1 hour and then filtering; And
And concentrating the supernatant obtained by centrifuging the filtrate to a concentration of 20 to 40 Brix under reduced pressure to obtain a concentrated liquid. The method for producing a fermented product of a two-stage fermented mushroom fruiting body according to claim 1, 8. The method of claim 7, wherein after the concentration step,
Drying the concentrate by lyophilization or spray-drying; And
Drying the precipitate by centrifuging the precipitate obtained by adding 5 to 10 times by weight of an organic solvent to the concentrate, cooling the mixture at 4 ° C for 24 hours, and lyophilizing the precipitate; &Lt; / RTI &gt; wherein the drying step selected from the group consisting of the following steps is further carried out. A fermented product of a mushroom fruiting body produced by the method according to any one of claims 1, 7 and 8. 10. The anticancer composition according to claim 9, which comprises a fermented product of a leaf mushroom fruiting body. 11. The method of claim 10,
Wherein the anticancer composition is a solid cancer or multiple cancer composition.

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