JP2010043017A - Hot-water extract from white to yellow deer-horn ganoderma lucidum, and cosmetics, health food and anticancer drug each comprising the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a hot-water extract comprising a substance having a physiological activity to humans, obtained from a fruit body of white to yellow Ganoderma lucidum, and cosmetics, health food and an anticancer drug each comprising the same. <P>SOLUTION: Disclosed are, (1) a hot-water extract comprising a substance having physiological activity to humans, obtained from a fruit body of white to yellow Ganoderma lucidum, (2) a hot-water extract according to (1), wherein the substance having physiological activity to humans has a growth-promotion action on a normal human fibroblast (NB1RGB), (3) a hot-water extract according to (1), wherein the substance having physiological activity to humans has a tyrosinase inhibitory action, (4) a hot-water extract according to (1), wherein the substance having physiological activity to humans has an antioxidative action, (5) a hot-water extract according to (1), wherein the substance having physiological activity to humans has an antitumor effect, and (6) health food of low bitterness hardly containing ganoderic acid or the like and comprising the above hot-water extracts. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a hot water extract containing a substance having a physiological activity on humans, obtained from a fruit body of white to yellow Ganoderma lucidum, and a cosmetic, health food, and anticancer agent containing the same It is about.

2. Description of the Related Art Conventionally, a deer antler reed 1 having a shape shown in FIG. 1 is known. This Kakuno Ganoderma has no umbrella (not open) and has a white top 2, but the fungus pattern 3 has a brown color (hereinafter referred to as a brown Kakuno Ganoderma).
The brown deer anteater is inoculated with hyphae of Ganoderma lucidum on a broad-leaved cut log and densely spread (spread), and then the cut log is placed in a housing having a height of 1 to 2 m. The fruiting body can be grown by embedding (laying down) the cut surface (Kiguchi) so as to be visible and maintaining the inside of the housing under the following conditions (A) and (B).
(A) After budding (budding) from embedding (budding), when the larva grows about 3 cm and its lower part (fungal pattern) begins to turn brown, the upper part of the housing should be shielded from light.
(B) Adjusting and maintaining the temperature of the space in the housing to 25 to 30 ° C. and the humidity to about 80 to 90%.

Brown deer antler can grow fruit bodies in substantially the same manner as in the cultivation method described above, even when using a fungal bed (artificial medium) that can be artificially produced instead of a broad-leaved cut raw tree.

The inventors have examined the various cultivation methods (conditions) because the form and color of ganoderma vary greatly depending on the cultivation method, and inoculated the mycelium of Ganoderma lucidum on the hardwood cut logs. After being densely grown (infested), the cut log is embedded (down) in a housing having a height of several tens of centimeters so that the cut surface (cutting end) can be seen, and the light shielding condition in the housing, By changing temperature and humidity, we succeeded in changing the fungus pattern of the Kakukaku Ganoderma to white or yellow, and cultivated ornamental white or yellow Kano Kakugakushi (Ganoderma lucidum) that can withstand lovers' appreciation. A patent application has already been filed (Patent Document 5).
In addition, even when using a fungal bed (artificial medium) that can be artificially produced instead of a broad-leaved cut raw wood, it has succeeded in cultivating almost the same white or yellow deer anteater (Patent Document 6). .

For example, the following patent documents are publicly known as methods for cultivating medicinal or ornamental ganoderma and deer ganoderma.
Shoko 57-39605 JP-A-11-146728 JP 2006-25765 A JP 2006-271298 A Japanese Patent Application No. 2006-343664 Japanese Patent Application No. 2007-074081

  The inventors have succeeded in the above-mentioned cultivation method, and white to yellow deer anteater (Ganoderma lucidum, hereinafter also simply referred to as deer antler) is rich in various physiologically active substances for humans. The present invention was completed by knowing that almost no ganoderic acid (a bitter component) contained in the brown ganoderma lucidum.

The present invention is constituted by the following claims 1 to 11.
[Claim 1] A hot water extract obtained from the fruit body of white or yellow Ganoderma lucidum and containing a substance having physiological activity for humans.
[Claim 2] The hot water extract according to claim 1, wherein the physiologically active substance has a growth promoting effect on human normal fibroblasts (NB1RGB).
[3] The hot water extract according to [1], wherein the physiologically active substance has a tyrosinase inhibitory activity.
[Claim 4] The hot water extract according to claim 1, wherein the physiologically active substance has an antioxidant action.
[5] The hot water extract according to [1], wherein the physiologically active substance has an antitumor effect.
[Claim 6] A cosmetic containing the hot water extract according to any one of claims 1 to 5.
[Claim 7] A health food containing the hot water extract according to any one of claims 1 to 5.
[8] An anticancer agent containing the hot water extract according to any one of [1] to [5].
[Claim 9] A health food containing the hot water extract according to any one of claims 1 to 5, which contains almost no ganoderic acids and has little bitterness.
[Claim 10] The normal human body according to any one of claims 1 to 8, wherein a fruit body of white or yellow deer anteater (Ganoderma lucidum) obtained by the cultivation method described in (A) to (C) below is used. Fibroblast (NB1RGB) growth promoter.
(A) Growing fruit bodies of white or yellow Ganoderma lucidum using a broad-leaved cut log.
(B) After inoculating a mycelium of Ganoderma lucidum on a broad-leaved cut log and densely spreading (spread), the cut log is buried in the soil in the housing to the extent that the cut surface (the end of the cut) can be seen ( Lay down.
(C) Grow a ganoderma fruit body while maintaining the inside of the housing under the following conditions (a) to (c).
(A) Covering the upper part of the housing with a semi-translucent object until the larval turf begins to change color after filling (budding), sprouting (sprouting), and the transmissivity is 65 to 85% To do so.
(B) After the sprout, before the larval turf turns brownish brown, the upper surface of the housing is covered with a light-shielding material to enhance airtightness and completely block light.
(C) Adjust and maintain the temperature of the space in the housing at 25 to 35 ° C. and the humidity at 90% or more.
[11] A normal human fiber according to any one of [1] to [8], wherein a fruit body of white or yellow Ganodermalucidum obtained by the cultivation method described in (A) to (C) below is used. Growth promoter for blast cells (NB1RGB).
(A) Growing fruit bodies of white or yellow Ganoderma lucidum by the fungus bed cultivation method.
(B) After inoculating hyphae (Ganoderma lucidum) hyphae on the fungus bed and allowing them to grow densely, the fungus bed is stored and held in the housing.
(C) The inside of the housing is maintained under the following conditions (a) to (b) to grow a ganoderma fruit body.
(A) After the fungus bed is stored in the housing, the upper part of the housing is covered with a semi-transparent object until the ganoderma lucidum is sprouted (germination) and the young ganoderma begins to change color. , Making the light transmittance 65-85%.
(B) After the sprout, before the larval turf turns brownish brown, the upper surface of the housing is covered with a light-shielding material to enhance airtightness and completely block light.
(C) Adjust and maintain the temperature of the space in the housing at 25 to 35 ° C. and the humidity at 90% or more.

According to the present invention, a hot water extract containing a substance having physiological activity with respect to humans can be easily obtained from the fruit body of white or yellow deer anteater which did not exist conventionally, and using this, It has the effect that cosmetics (skin beautification, whitening) having various effects, health foods (powder, beverages), and anticancer agents can be obtained.
In addition, a hot water extract containing a substance having physiological activity for humans obtained from the fruit body of white to yellow deer antaceae according to the present invention is composed of ganoderic acids (bitter substances) which are components of conventional ganoderma. ) Is hardly contained, so that it can be used to obtain health foods (powder, beverages) having various effects (beautifying skin, whitening, antioxidant, anticancer).

Using the same tree species and fungus species, white kakukaku reishi (white) and kakukaku reishi (brown) were cultivated, and the general components and the amount of β-glucan were measured. The results are shown in Table 1 (Japan Food Analysis Center No. 50807368-001-004).
Note that β-glucan was measured by an enzymatic method.

<Sample preparation method for physiological activity measurement>
The dried product of the fruit body of white or yellow deer antler was finely pulverized, 20 times the amount of distilled water was added, treated with an autoclave at 105 ° C. for 60 minutes, and extracted with hot water.
The analysis results of the extract are shown in Table 2.

<Measurement of effects on normal human fibroblast proliferation>
In our skin, there are fibroblasts at the bottom of the epidermis, which keeps the skin elastic, such as collagen and elastin, and produces substances that are the source of elasticity. It is said that these fibroblasts decline with age, the ability to produce collagen and elastin decreases, the moisture is lost, the texture collapses, and wrinkles are caused.

For the measurement sample, the obtained extract (Table 1) was filtered and further sterilized by filtration with a 0.2 μm filter, and then prepared to 100 μg / ml (1 ml of sample corresponds to 100 μg of dried fruit body) and used for the test. Based on the results of the preliminary test, the proliferation rate of human normal fibroblasts in 24 hours of culture was measured for the white to yellow deer anteater extract by the following procedure. For the measurement, CellTiter96 Aqueous One Solution Cell Proliferation Assay (manufactured by Promega Corporation) was used.
The outline of the measurement method is as follows.
<1> Human normal fibroblasts (NB1RGB) were precultured.
<2> Pre-culture, cells in stationary phase were trypsinized to prepare a cell suspension, and the number of cells was counted.
<3> The cells were seeded in a 96-well microplate at 1 × 10 ⁵ cells / well.
<4> Cultured in a serum-free medium for 16 hours.
<5> The medium was replaced with an evaluation sample-added medium.
(A) Control (negative control, no addition, with cells)
(B) Hot water extract of dried white deer ginseng fruit body (c) Hot water extract of dried brown ganoderma berries <6> Cultured in a serum-free medium for 24 hours.
<7> After collecting the medium, the number of cells was measured by the MTS method using CellTiter 96R AQueous One Solution Cell Proliferation Assay (manufactured by Promega Corporation).
Table 3 shows the measurement results.

As described above, the cell growth period was set to 24 hours, and cell proliferation in this period was examined.
As a result, the hot water extract of the dried deer ergot fruiting body showed a sufficient human normal fibroblast proliferation rate (133%).
Therefore, the hot water extract of white deer horn reed fruit bodies can be used as a cosmetic raw material for improving self-regeneration ability.
In addition, although the morphological change of the cell was observed with the optical microscope, the characteristic morphological change was not seen by sample addition.

<Measurement of tyrosinase inhibitory activity>
Tyrosinase is an enzyme that oxidizes tyrosine, an amino acid, to produce melanin.
The sample used the extract of Table 1 as it was without adjusting the concentration.
This test method is a method for testing the presence or absence of a whitening effect by measuring the inhibition rate of tyrosinase activity.
Mushroom-derived tyrosinase enzyme (50,000 units) (manufactured by Sigma Chemical Co., Ltd.) was used for the measurement.
The outline of the measurement method is as follows.
<1> A sample tube mixed with 1.0 ml of McIlvaine buffer (pH 6.8), 1.0 ml of tyrosine solution, and 0.9 ml of sample solution was heated at 30 ° C. for 10 minutes.
<2> 0.05 ml of 918 units / ml tyrosinase was added and further heated to 30 ° C. for 10 minutes.
<3> 0.05 ml of 1M sodium azide was added to stop the reaction.
<4> Absorbance at 475 nm was measured.
Table 4 shows the measurement results.
Table 4 shows the inhibition rate of each sample calculated from the value of Control.

From the measurement results in Table 4, the hot water extract of the dried deer ergot fruiting body was found to inhibit tyrosinase activity.
Factors of stain include changes in hormone balance, rough skin, ultraviolet rays, aging, etc., all of which are related to ultraviolet rays. When exposed to UV rays, keratinocytes, which occupy most of the epidermis, protect themselves from UV-stimulated epidermal cells, so that signaling substances (endolysene and phospholipase) are sent to melanocytes that produce melanin. In response, tyrosinase produces melanin. In the living body, the amino acid tyrosine becomes dopa by the action of tyrosinase enzyme, and then becomes dopaquinone. An oxidation reaction proceeds from dopaquinone to form melanin. If the reaction can be suppressed / inhibited at the beginning of this melanin formation pathway, the amount of melanin produced can be greatly reduced. Since the tyrosinase activity inhibition was recognized in the hot water extract of the dried fruit body of Shirakakaku Reishi, it can be expected to suppress melanin production. Based on the above, it was considered that the hot water extract of the dried white deer kakushiba fruit body can be expected as a whitening cosmetic raw material.

<Antioxidation test (DPPH radical scavenging ability evaluation test)>
Antioxidant activity was determined by a DDPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging ability evaluation test.
When free radicals are generated by oxidative stress in vivo, it is said to cause various diseases such as aging, cancer, and arteriosclerosis, and substances that supplement free radicals are useful for preventing such diseases.
The sample used the extract of Table 1 as it was without adjusting the concentration.
The outline of the measurement method is as follows.
<1> A mixed solution (1: 1: 1) of DDPH (1,1-diphenyl-2-picrylhydrazyl) in ethanol (4 μM), MES buffer (200 mM, pH 6.0), and 80% ethanol was prepared.
<2> Six test tubes were prepared for each sample, and distilled water (300-a) μl was added. Said a shows the sample amount added by <3>. Here, 300, 270, 240, 180, 120, and 60 μml were added to six test tubes, respectively.
<3> A sample (a = 0, 30, 60, 120, 180, 240) was added to the test tube and stirred.
<4> The reaction solution of <1> was added to each test tube and allowed to react with stirring.
<5> The above reaction solution was placed in a cell and set in a spectrophotometer. Absorbance at 520 nm was measured 20 minutes after adding the extract.
<6> A standard Trolox solution (0.2 mM) was prepared, and the operations <1> to <5> were performed.
<7> A plot of the sample and Trolox was plotted with the absorbance on the vertical axis and the added amount (μl) on the horizontal axis. Next, in the range in which linear absorbance decrease continues (standard: linearity R2 = 0.9999, sample: linearity R2 = 0.975 or more), the amount of change in absorbance per unit addition is calculated and compared. As a result, the mass of the antioxidant contained in the sample (corresponding to Trolox) was calculated.
Table 5 shows the measurement results.

From the above, it can be seen that the hot water extract of dried deer ergot repellent substance has scavenging activity against various radicals.
Therefore, it can be used as a raw material for health foods useful for preventing diseases caused by free radicals.

<Anti-tumor test (cell growth inhibition test and apoptosis induction test)>
(A) Cell Proliferation Inhibition Test <1> The sample was used as it was without adjusting the concentration of the extract of Table 1.
<2> Cell culture was performed according to the following method.
Cell proliferation suppression test and apoptosis induction test (in vitro) were performed using human myeloid leukemia-derived cells (HL-60 cells).
Human myeloid leukemia-derived cells (HL-60 cells) were RPMI supplemented with 10% fetal bovine serum (FBS; Funakoshi), penicillin (100 U / ml), streptomycin (100 μg / ml) (penicillin-streptomycin mixture; SIGMA). Culturing was performed using 640 medium (GIBCO) in the presence of 37 ° C. and 5% CO 2. Cells were diluted 5-6 times once every 3-4 days and subcultured.
The cell count was stained with 0.5% trypan blue (Wako) using a hemocytometer and measured under a microscope.
All tests were performed using cells in logarithmic growth phase.
<3> The cell growth inhibition test was performed as follows.
A suitable amount of Hl-60 cells in the logarithmic growth phase was collected in a 15 ml centrifuge tube and centrifuged at 1.000 rpm for 5 minutes. The medium was removed from the centrifuged tube using an aspirator, and the number of cells was diluted and adjusted using a 50 ml centrifuge tube so that the medium was warmed to 37 ° C. to 5 to 6 × 10 5 cells / ml.
900 μml each of the prepared cell suspension was seeded in a 24-well plate. As a control, 100 ml of distilled water and each sample were added to each well and allowed to stand in a CO 2 incubator at 37 ° C.
After 24 hours, the cells were stained with 0.5% trypan blue, the number of cells was counted using a hemacytometer, and the cell growth inhibition rate was calculated.
Table 6 shows the measurement results.

  According to Table 6, it can be seen that the hot water extract of the dried white deer ganshi fruit body exhibits a high cell growth inhibitory effect.

(B) Confirmation of apoptosis-inducing cells by Giemsa staining An appropriate amount of HL-60 cells in the logarithmic growth phase was recovered in a 15 ml centrifuge tube and centrifuged at 1000 rpm for 5 minutes. Remove the medium from the centrifuged tube using an aspirator in a clean bench and dilute the number of cells using a 50 ml centrifuge tube so that the medium is warmed to 37 ° C to 5-6 x 105 cells / ml. did.
Dispense 900 μl of the prepared cell suspension into a 24-well plate, and use distilled water (Control (DW)) as a control for each well and each sample (G2: Shirakakaku Reishi, G3: Brown spirit). Turf) was added in an amount of 100 μl, and allowed to stand in a CO 2 incubator at 37 ° C.
The cells taken out after 24 hours were collected in Spitz tubes. The same amount of Carnoy's fixative (methanol: acetic acid = 3: 1 mixed solution) was filled therein. After allowing to stand at room temperature for 10 minutes, the mixture was gently tilted up and down and allowed to stand for another 10 minutes. After centrifuging at 1500 rpm for 10 minutes and removing the supernatant, 2 ml of Carnoy's fixative was added and suspended, and allowed to stand at −20 ° C. overnight. The next day, the mixture was centrifuged at 1500 rpm for 10 minutes, and the supernatant was removed. Then, the suspension was suspended in 200 μl of ice-cooled Carnoy's fixative and dropped onto a slide glass. When dried, it was stained with Giemsa staining solution (Wako diluted 100-fold and used) for 20 minutes and washed with distilled water. The cell morphology was confirmed under a microscope, and photography was performed.

Morphological features in apoptotic cells include nuclear condensation.
As a result of the above test, the confirmed cell morphology is shown in FIG. In confirming the cell morphology in Giemsa staining, many apoptosis-inducing cells could be confirmed. In addition, the proportion of apoptosis-inducing cells was confirmed at a rate almost equal to the cell growth inhibition rate.
From the results of the cell growth inhibition test and the confirmation test of apoptosis-inducing cells, Shirakakaku Reishi's substance dry matter hot water extract suppressed the growth of HL-60 cells at a high rate, and the suppression was due to cell death due to apoptosis. It was inferred that there was.

<Quantitative determination of ganoderic acid A>
<1> The sample used was the extract described in paragraph 0010 (1 ml of the sample corresponds to 100 μg of dried fruit body).
<2> Liquid chromatography was performed under the following conditions.
Column: ODS column (SINSEIDO CAPCELL PAC C18), 35 °
Mobile phase: acetonitrile / water / acetic acid = 35/65/1
Flow rate: 1.0 ml / min Detector: UV 254 nm

<3> Analytical result Conventional hot water extract of brown deer anteater contained 140 (mg / 100 g reindeer fruit body) or more ganoderic acid A. In the hot water extract, 10 mg or more of ganoderic acid A could not be found.

<4> Sensory test results About the bitterness of the samples in Table 1, when the sensory test was performed using 10 panels, the following results were obtained.

Shirakakaku Reishi no bitterness-8 people slightly bitter -2 people bitterness-0 people
No bitterness-1 person
Slightly bitter-3 people
-6 bitter taste

From the above results, the hot water extract containing a substance having physiological activity against humans obtained from the fruit body of white or yellow deer antrum according to the present invention is a bitter substance, like conventional brown ganoderma Is hardly contained. Therefore, it is possible to obtain a health food (powder, beverage) having various effects (beautifying skin, whitening, antioxidation, anticancer) even without bitterness.

The Shirakakaku Reishi used in the present invention has the following properties (see Patent Document 5 and Patent Document 6).
(A) The total length (height) is 25 to 40 cm and does not branch.
(B) The tip (the part corresponding to the fungus umbrella) is white (c) The fungus pattern is white to yellow (d) The diameter of the fungus pattern is approximately 10.0 mm (upper) to 20.0 (lower) mm

Next, a method for cultivating white deer antler used in the present invention will be described.
(1) When using broad-leaved cut logs (i) The broad-leaved tree used is excellent in kunugi or ragweed, but Japanese oak, mizunara, abemaki, sakura, beech, fake acacia, etc. can also be used sufficiently.
(B) A known method can be used to inoculate a reticulated mycelium on a hardwood cut raw tree so as to densely spread (spread). For example, Japanese Patent No. 3553005 or Japanese Patent Application Laid-Open No. 2002-84884 (both Both methods are simpler and more reliable if the method described in the inventor's invention is used.
(C) A hardwood cut raw wood having a diameter of 15 to 25 cm and a height of about 10 to 16 cm is used. The size of the raw wood is not necessarily limited to this size, but this range of raw wood is convenient for work and management.

(D) Mountain sand with good water drainage is suitable for the soil where the cut log, which is densely spread (spread) with reishi mycelia, is buried. The embedding should be almost up to the height of the raw wood, and the sand on the cut (cutting) surface should be removed.
(E) The distance between the left and right timbers to be embedded should be in the range of 7 to 15 cm (9 to 10 per square meter). If it is narrower than this range, the growth of ganoderma will worsen, and if it exceeds this range, the fungus pattern will be yellow (browning) and the color (browning) will tend to increase.
(F) The height of the inside of the housing is preferably 30 to 65 cm, more preferably 35 to 45 cm. If it is lower than this range, the growth of ganoderma is worsened, or the ganoderma grows sideways and the appearance deteriorates.
On the other hand, if it exceeds this range, the fungus pattern will be yellowed (browned) beyond yellow, or the number of colored (browned) will tend to increase.
The distance between the log and the height of the inside of the housing determines the space (volume) where the reishi generated from a single log grows. The size of this space is sensitive to the color of the reishi. Affects.
That is, as space increases (if it is too large), coloring increases.
The results of detailed tests using several logs showed that the carbon dioxide concentration in the housing was 0.1% (normal air concentration: 0.03%) or more, and that the fungus pattern was browned. In order to secure a space of this carbon dioxide concentration, the ventilation is limited, and the distance between the (raw wood) and the height (inside the housing) are optimal.

(G) Next, a configuration example of the housing is as follows.
The entire structure is a side plate that surrounds the periphery in a box shape and a canopy that covers the upper portion of the side plate, and a space with a height of 35 to 45 cm is secured therein. The side plate and canopy should be made of heat-insulated material so that they can be completely shielded from light and sealed.
As a heat insulating material to be used, for example, the following board is used.
Inside: Insulating material (Styrofoam or extruded polystyrene foam (trade name: Styrofoam): 9-30 mm)
Outside: Plywood (Coated outside with a light-shielding film if necessary): 5-10 mm
(H) The canopy is not opened or closed except when watering is usually performed once every 3 to 5 days in order to avoid rapid changes in temperature and humidity and to maintain the increased carbon dioxide concentration due to the growth of ganoderma.
In general, watering is performed at night, and the canopy is not opened or closed during the day to prevent direct sunlight from being inserted.

(I) It is necessary to maintain the temperature in the housing at 25 to 35 ° C. and the humidity at 90% or more, more preferably 95% or more. This condition can be achieved, for example, by changing the thickness of the heat insulating material of the canopy (if the temperature is too low, it is thinned to capture the heat of sunlight into the housing, and if it is too high, it is thickened to block the heat of the sunlight) ) And by changing the amount and frequency of watering.
(Nu) The time when the housing is completely shielded from light by a canopy or the like is preferably just before the larval turf discolors, and this time usually comes around 5 to 7 days after sprouting.

(L) The semi-transparent object used in (c) of (C) of claim 9 (the one covering the upper part of the housing during the period when the larval turf begins to change color after budding and germination) A sheet-like object that partially blocks sunlight.
In addition, the object can be achieved even if a semi-transparent object is laminated on a translucent film (sheet) by covering a part of the translucent film (sheet).
If the inside of the housing is not maintained under the condition (C) before sealing with the canopy, discoloration of the ganoderma will be accelerated or the growth (growth) of the ganoderma will be worsened.

(Wo) When cultivating a large amount (for example, 100 strains or more) simultaneously in the same space, outdoor cultivation (sunlight irradiation) is performed.
For this purpose, in mid-May, it is better to bury (cut down) a cut log, which is densely spread (spread) with reishi hyphae, and harvest from mid-July to early August.
In this period, the temperature is maintained at 25 to 35 ° C. and the humidity is maintained at 90% or more by appropriately watering the space and changing the thickness of the heat insulating material of the canopy.

(2) When using a fungus bed (i) The fungus bed (medium) is produced mainly from hardwood sawdust (or chips) and rice bran (or bran). As a broad-leaved tree, Kunugi or Naragashi is excellent, but Japanese oak, Mizunara, Abemaki, Sakura, Beech, Fake Acacia, etc. can be used sufficiently.

(B) In order to inoculate the mycelium with the ganoderma hyphae and densely spread (spread), a known method can be used. For example, Japanese Patent No. 3553005 or Japanese Patent Application Laid-Open No. 2002-84884 (both If the inoculum is cultured using the method described in the invention of the present inventor, it is more simple and reliable.
It usually takes 120 days or more to inoculate the mycelium with the ganoderma hyphae and allow them to grow dense.

(C) The container used for the preparation of the fungus bed has a wide opening at the top, and since it is necessary to sterilize the medium inside, a heat-resistant container that can withstand heat sterilization is preferable. For example, glass and plastic containers (bottles, bags, etc.) can be used.
Although the size of the container is not particularly limited, a container having a bottom area of 80 to 200 cm ² (diameter 10 to 16 cm) and a height of about 10 to 15 cm is convenient in terms of work and management.

(D) It is preferable to regulate the arrangement of the fungus bed when the fungus bed in which the mycelium of reishi is densely (spread) is stored and held in the housing as follows.
a) Limiting the number of bacteria beds stored and arranged in the housing so that the sum of the bottom areas of the bacteria beds to be stored occupies 20 to 30% of the bottom area in the housing, with an appropriate interval Place the fungus bed.
b) Take no space under the fungus bed.
c) If the fungus bed is stored and arranged too densely, the growth of the ganoderma will worsen, and if it is stored and arranged too loosely, the fungus pattern of the ganoderma will exceed the yellow color (browning), or coloring ( There is a tendency for more to be browned.

(E) When the fungus bed is stored and arranged in the housing, it is preferable to secure a space of 20 to 55 cm, more preferably 25 to 40 cm above the fungus bed. If it is lower than this range, the growth of ganoderma is worsened, or the ganoderma grows sideways and the appearance deteriorates.
On the other hand, if it exceeds this range, the fungus pattern will be yellowed (browned) beyond yellow, or the number of colored (browned) will tend to increase.
The space between the fungus beds and the height of the inside of the housing (space to be secured above the fungus beds) determines the space (volume) in which the reishi generated from one fungus bed grows. The size of this space is sensitive to the coloration of Ganoderma.
That is, as space increases (if it is too large), coloring increases.
As a result of detailed tests using several fungi beds, browning of the fungus pattern by increasing the carbon dioxide concentration in the housing to 0.1% (normal air concentration: 0.03%) or more In order to secure a space with this carbon dioxide concentration, the ventilation is limited, and the above-mentioned (bacteria bed) interval and (height bed top) height (space) are optimal.

(F) Next, a configuration example of the housing is as follows.
The entire structure is a side plate that surrounds the periphery in a box shape and a canopy that covers the top of the side plate, and a space of 35 to 50 cm in height is secured in the interior. The side plate and canopy should be made of heat-insulated material so that they can be completely shielded from light and sealed.
As a heat insulating material to be used, for example, the following board is used.
Inside: Insulating material (Styrofoam or extruded polystyrene foam (trade name: Styrofoam): 9-30 mm)
Outside: Plywood (Coated outside with a light-shielding film if necessary): 5-10 mm

(G) The canopy is normally watered once every 3 to 5 days (to maintain the humidity inside the housing) in order to avoid sudden changes in temperature and humidity and to maintain the increased carbon dioxide concentration due to the growth of ganoderma Do not open or close except when.
In general, watering is performed at night, and the canopy is not opened or closed during the day to prevent direct sunlight from being inserted.

(H) It is necessary to maintain the temperature in the housing at 25 to 35 ° C. and the humidity at 90% or more, more preferably 95% or more. This condition can be achieved, for example, by changing the thickness of the heat insulating material of the canopy (if the temperature is too low, it is thinned to capture the heat of sunlight into the housing, and if it is too high, it is thickened to block the heat of the sunlight) ) And by changing the amount and frequency of watering.
With the housing configuration as described above, it was measured that the temperature inside the housing would rise and fall almost certainly within the range of 25 to 35 ° C. (temperature difference 7 to 9 ° C.) during one day.

  (I) The time when the housing is completely shielded from light by a canopy or the like is preferably just before the larval turf is discolored, and this time usually comes around 5 to 7 days after sprouting.

(Nu) The semi-translucent object used in (a) of (C) of claim 10 (the one covering the upper part of the housing during the period when the larval turf begins to change color after budding and germination) A sheet-like object that partially blocks sunlight.
In addition, as a semi-transparent object used for achieving the conditions of the present invention, a sheet (for example, net-like or bowl-like) covering a part of the translucent film (sheet) is overlaid. Even the purpose can be achieved.
If the inside of the housing is not maintained under the condition (C) before sealing with the canopy, discoloration of the ganoderma will be accelerated or the growth (growth) of the ganoderma will be worsened.

(L) When cultivating a large amount (for example, 100 strains or more) simultaneously in the same space, outdoor cultivation (sunlight irradiation) is performed.
For this purpose, it is recommended to place a fungus bed in which the hyphae of Ganoderma lucidum are densely spread in the housing in mid-May and harvest it from mid-July to early August.
In this period, the temperature is maintained at 25 to 35 ° C. and the humidity is maintained at 90% or more by appropriately watering the space and changing the thickness of the heat insulating material of the canopy.

Shirakakaku Reishi used for the test of the present invention was obtained by the following cultivation method.
2 is installed on the ground in Neba Village, Shimoina District, Nagano Prefecture. The specifications of the housing are as follows.
Size (inside): 710 cm (length) x 150 cm (width) x 40 cm (height)
Side plate 5: Styrofoam 7 (inside, 10 mm) is attached to plywood plate 6 (outside, 7 mm) Canopy 9: Styrofoam 7 (inside, 20 mm) is attached to plywood plate 6 (outside, 7 mm) Ground soil (mountain sand) 13 : About 20cm of well-drained mountain sand was laid under the ground.

The cultivation of Shirakakaku Reishi has gone through the following process.

Preparation of raw wood →→ Inoculation and culture of inoculum →→ Hide down →→ Cover (covered with canopy) →→ Growth →→ Harvesting

B) Log preparation Logs were cut from November to February. A portion having a diameter of 15 cm to 25 cm (average 19 cm) was cut into a length of about 13 cm to obtain a cut log 4. These were packed in a bag and sterilized in an atmospheric sterilization pot for about 2.5 hours.
B) Inoculation and culture of inoculum The inoculation and culture of the reindeer inoculum to the cut log 4 were carried out according to the method of Japanese Patent No. 3553005.
The culture period is 120 to 150 days, and 30 days after inoculation is cultured in a simple culture room set at a temperature of 25 ° C. to 27 ° C. and a humidity of 40 to 60%. From 4 onwards, it was managed in the natural environment (temperature).

C) Saddle down
A cutwood 4 of Knugi that had been infested with the reed mycelium was buried in the mountain sand 13 in the housing 12 so that the cut surface (the end) was visible. The mutual distance between the cut logs 4 was about 10 cm (mid-May).
D) Next, the upper part of the housing 12 was covered with a translucent sheet (polyethylene) 10 and a light-shielding net (black, bowl-shaped) 11 so that the transmissivity was 65 to 85%.
E) Lid Ganoderma sprouting (cutting) around 10 days after lying down. About one week later, the upper portion of the housing 12 was covered with the canopy 9, completely shielded from light, and sealed.
The canopy 9 was opened and closed (watering) at night, once every 3 to 5 days, with reference to temperature (30 ° C.) and humidity (95% or more).

(F) Growth Ganoderma grew smoothly and had the following characteristics in mid-July.
(A) The total length (height) is 25 to 40 cm and does not branch.
(B) The tip (the part corresponding to the fungus umbrella) is white (c) The fungus pattern is white to yellow (d) The fungus pattern has a diameter of approximately 10.0 mm (upper part) to 20.0 (lower part) mm
(G) Harvesting Harvesting took from the sand 13 the cut log 4 with white or yellow fungus from the beginning to the middle of August.

It is a perspective view of Kazuno Ganoderma. It is a perspective view which shows the housing for Kazuno Ganoderma cultivation. It is a partial view explaining the structure of the side plate of a housing, a translucent sheet | seat (polyethylene), and a light-shielding net | network. It is a fragmentary perspective view which shows the state which coat | covers a housing with a canopy. It is a front view which shows the structure of a canopy It is a fragmentary perspective view which shows the growth state of white thru | or yellow deer antler. It is a figure which shows the form change of apodosis.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Kazuno Ganoderma 2 Top part 3 Fungi pattern 4 Cutting log 5 Side board 6 Plywood 7 Styrofoam 8 Splint 9 Canopy 10 Translucent sheet 11 Light shielding net 12 Housing 13 Ground soil (mountain sand)
14 Prop

Claims (11)

A hot water extract obtained from the fruit body of white or yellow Ganoderma lucidum and containing a substance having physiological activity on humans. The hot water extract according to claim 1, wherein the substance having physiological activity has an effect of promoting the growth of human normal fibroblasts (NB1RGB). The hot water extract according to claim 1, wherein the physiologically active substance has a tyrosinase inhibitory activity. The hot water extract according to claim 1, wherein the substance having physiological activity has an antioxidant effect. The hot water extract according to claim 1, wherein the physiologically active substance has an antitumor effect. Cosmetics containing the hot water extract according to claim 1. A health food containing the hot water extract according to claim 1. The anticancer agent containing the hot water extract of Claims 1-5. A health food containing the hot water extract according to claims 1 to 5, which contains little ganoderic acid and has little bitterness. The human normal fibroblast (NB1RGB) according to claim 1 to 8, wherein the fruit body of white or yellow deer anteater (Ganoderma lucidum) obtained by the cultivation method described in (A) to (C) below is used. ) Growth promoter.
(A) Cultivation of the fruit body of white or yellow Ganoderma lucidum using broad cut wood.
(B) After inoculating a mycelium of Ganoderma lucidum on a broad-leaved cut log and densely spreading (spread), the cut log is buried in the soil in the housing to the extent that the cut surface (the end of the cut) can be seen ( Lay down.
(C) Grow a ganoderma fruit body while maintaining the inside of the housing under the following conditions (a) to (c).
(A) Covering the upper part of the housing with a semi-translucent object until the larval turf begins to change color after filling (budding), sprouting (sprouting), and the transmissivity is 65 to 85% To do so.
(B) After the sprout, before the larval turf turns brownish brown, the upper surface of the housing is covered with a light-shielding material to enhance airtightness and completely block light.
(C) Adjust and maintain the temperature of the space in the housing at 25 to 35 ° C. and the humidity at 90% or more. The human normal fibroblast (NB1RGB) according to claim 1 to 8, wherein the fruit body of white or yellow Ganodermalucidum obtained by the cultivation method described in (A) to (C) below is used. Growth promoter.
(A) Growing fruit bodies of white or yellow Ganoderma lucidum by the fungus bed cultivation method.
(B) After inoculating hyphae (Ganoderma lucidum) hyphae on the fungus bed and allowing them to grow densely, the fungus bed is stored and held in the housing.
(C) The inside of the housing is maintained under the following conditions (a) to (b) to grow a ganoderma fruit body.
(A) After the fungus bed is stored in the housing, the upper part of the housing is covered with a semi-transparent object until the ganoderma lucidum is sprouted (germination) and the young ganoderma begins to change color. , Making the light transmittance 65-85%.
(B) After the sprout, before the larval turf turns brownish brown, the upper surface of the housing is covered with a light-shielding material to enhance airtightness and completely block light.
(C) Adjust and maintain the temperature of the space in the housing at 25 to 35 ° C. and the humidity at 90% or more.