JPS633576B2 - - Google Patents

Description

[Detailed description of the invention]

TECHNICAL FIELD The present invention relates to an extract of Mantis edulis, a method for producing the same, and a health food containing the extract as an active ingredient. Stone mushroom is a basidiomycete that belongs to the order Polygonum and the family Sarnocotyaceae.It is also known as Ganoderma, and has been prized as an excellent herbal medicine in China since ancient times. In recent years, much research has been carried out in Japan to scientifically elucidate the active ingredients of Cinnamon mushroom, and its anticancer, antihypertensive,
Its anti-hyperlipidemic effects are gradually becoming clearer, and various attempts have been made to isolate the active ingredient and use it as a medicine, and to use the mushroom itself as a so-called health food. However, Cinnamon mushrooms originally grow very rarely in nature even in China, making them extremely difficult to obtain and very expensive. Recently, in our country, it has become possible to artificially cultivate Cinnamon mushrooms and they are relatively easy to obtain, but they are still expensive and require a long period of 2 to 5 years to grow. In addition, there is a problem in that many of the artificially cultivated stone mushrooms contain different ingredients compared to natural ones. In view of these circumstances, the present inventors conducted extensive research in order to easily and inexpensively obtain a stone mushroom that has the same or similar components as natural ones, and unexpectedly found a specific A liquid medium having a composition of We found that the components were also accumulated in the culture solution, and that extracts obtained by concentrating these components or extracting them with specific solvents are suitable for health foods. As mentioned above,
Artificial cultivation of Cinnamon lucidum has already been carried out, but this is the cultivation of the fruiting bodies of Cinnamon lucidum, and as in the present invention, there are examples of efficient liquid culture of Cinnamon mycelium, the obtained culture solution, and mycelium extract. , and health foods using it have not been found so far. Thus, the present invention is directed to a novel method in which a culture obtained by liquid-cultivating C. chinensis mycelium in a liquid medium with a specific composition, or a culture obtained by extracting the separated mycelium with a specific solvent, or a concentrated culture solution from which the mycelium has been separated. The present invention provides a pine mushroom extract, a method for producing the same, and a health food containing the extract as an active ingredient. According to the present invention, by liquid-cultivating C. albacore mycelium in a liquid medium containing 0.2 to 10 W/V% glucose and 0.2 to 2 W/V% wheat germ as essential components, it is possible to achieve high yields in a short period of time and at low cost. , it is possible to obtain a C. latinum mycelium containing components similar to those contained in the natural C. latinum fruiting body, and the components can also be accumulated in the culture solution, and after the completion of the cultivation,
extracting the culture, i.e., a mixture of mycelium and culture solution or separated mycelium, with a solvent selected from the group consisting of water, alcohol, chloroform, ether, ethyl acetate, benzene, hexane and mixed solvents thereof; By concentrating the culture solution from which the mycelium has been separated, the Physcomitrium extract can be obtained. The obtained extract can be made into a health food in a known form as it is or according to a conventional method, and can be used in the same way as a health food using natural Physcomitrella edulis fruiting bodies. That is, in order to obtain the Cinderella extract of the present invention, first, the mycelia of C. monocytogenes are cultured in a liquid medium containing glucose and wheat germ as essential components. The glycose and wheat germ used as the medium components may be any of those commonly available as medium components. Glucose is 0.2 to 10 W/V%, preferably 1 to 8 W/V% based on the total amount of the medium.
Use at a ratio of The amount of glucose is 0.2W/V%
Even if it is lower or exceeds 10 W/V%, the mycelium yield will decrease. Wheat germ is 0.2 to 2 W/V%, preferably 0.5 to 1.0 W/V, based on the total amount of the medium.
Used in percentages. The amount of wheat germ is also 0.2W/V%
If lower, mycelium yield will decrease, and 2W/
If it exceeds V%, it will be economically disadvantageous. Particularly, in the present invention, it is preferable that the weight ratio of wheat germ:glucose in the liquid medium is 1:1 to 8, thereby improving the yield of Mosquidium mycelium. Optionally, the liquid medium may contain mineral components such as salts such as phosphorus, manganese, magnesium, calcium, iron, etc., as well as other grain germs, rice bran, etc.
Other nutritional ingredients such as corn steep liquor, vitamins, nucleic acids, starch, amino acids, yeast extract, peptone, etc. may be added as appropriate. The liquid medium can be prepared according to a conventional method, for example, by adding each predetermined component to sterile water, dispersing it,
Dissolve. The resulting medium is usually kept at 120-130°C.
After being sterilized for 15 to 30 minutes, it is used for culturing the C. chinensis mycelium. Cultivation of C. chinensis mycelium is carried out under aerobic conditions by inoculating an appropriate amount of seed mycelia into the liquid medium. The seed hyphae to be used may be any seed fungi belonging to the Basidiomycete order, the order Aridaceae, and can be obtained from, for example, Kawamura Shiitake Research Institute (1-1-11 Aoba-cho, Fujieda City, Shizuoka Prefecture). Usually, the amount of seed mycelia inoculated is about 5 to 10 mg/100 ml of medium, and under stirring at 200 to 300 rpm, the temperature
By culturing in the dark for 7 to 21 days at 25 to 30°C and an aeration rate of 0.5 to 3.0 VVm, a high yield of C. monocytogenes mycelium having components similar to those contained in the natural C. monocytogenes fruiting body can be obtained. , the components can be accumulated in the culture medium (for example, according to this culture, compared to potato dextrose broth (PDB) medium used for conventional culture of seed for artificial cultivation of C. even mycelial yields can be achieved). After culturing, the resulting culture (mixture of mycelium and culture solution) is extracted directly, the mycelium separated from the culture is extracted with a solvent, or the remaining culture solution after the mycelium is separated from the culture is concentrated. The extract of the present invention is obtained. Separation of mycelium from the culture can be carried out according to conventional methods such as filtration and centrifugation. For extraction of the culture or isolated mycelium with a solvent, if necessary, after cutting the mycelium appropriately, add water,
Alcohol (e.g. methanol, ethanol,
n-butanol, etc., chloroform, ether,
Ethyl acetate, benzene, hexane and mixtures thereof Solvents (e.g. 50% ethanol aqueous solution, 80%
% methanol aqueous solution, etc.). The extraction generally includes
Using a solvent in an amount of about 10 to 100 times the weight of the material to be extracted, the extraction is carried out at a temperature of 15° C. to the boiling point of the solvent for 1 to 24 hours. It is usually sufficient to perform this operation one to three times. The obtained extract is preferably heated under reduced pressure.
Concentrate at 40-50℃, if necessary, 40-50℃ under reduced pressure.
The extract of the present invention is obtained by drying with . Further, the remaining culture solution from which the mycelium has been separated can be concentrated in the same manner as the above-mentioned extraction solution, and if necessary, dried in the same manner to obtain the L. chinensis extract of the present invention. The resulting extract is generally in the form of a brown to brown paste to powder, with a slightly sweet taste, no unpleasant flavor, and a fragrant wheat germ flavor. It is amorphous and slightly soluble in water. The extract is relatively stable up to about 60° C., and as described later, contains components that are very similar to those of the natural fruiting body of L. chinensis. In addition, the extract showed positive results in all of the ninhydrin reaction, α-naphthol reaction, Fehring reaction, phenylhydrazine reagent reaction, Anthrone-sulfuric acid reagent reaction, Biuret reaction, and ferric chloride reagent reaction, and showed positive results in the Elson-Morgan reaction. Indicates a false positive. From these properties and the results of various spectral analyzes and chromatographic analyses, it is considered that the L. chinensis extract of the present invention contains polysaccharides and glycoproteins as its main components. The attached Figures 1 to 3 show gas chromatograms, thin-layer chromatograms, and high-performance liquid chromatograms of representative examples of the Cinnamon extract of the present invention (produced according to Example 1 described later and re-extracted with ethyl acetate). show. The conditions for each chromatography are as follows. Gas chromatography Model used: Shimadzu GC-7A; Column: 2
%OV-1, Uniport HP (60-80 mesh),
3mm x 200cm; Column temperature: 8 minutes at 150℃,
Then, the temperature rises to 230℃ in 80 minutes; vaporization chamber temperature: 280
°C; Detector: FID. In addition, the sample was prepared according to the usual method.
Converted to TMS. Thin layer chromatography plate: Merck silica gel; developing solvent:
Chloroform-methanol (9:1); Color development: I ₂
and ultraviolet rays (the spots indicated by dotted lines in Figure 2 are
Color develops with I ₂ ; spots shown by solid lines develop color under an ultraviolet lamp, and strongly develop with I ₂ ). High performance liquid chromatography Model used: Shimadzu LC-3A; Detector:
SCP-2A; Column: YMC-PACK A-311, 6
×100mm (Yamamura Chemical Research Institute); Flow rate: 1.0ml/min;
Mobile phase: 35% acetonitrile/0.2M acetate buffer (PH3.5); internal standard: ethyl p-aminobenzoate; detection wavelength: 254 nm. In FIGS. 1 and 2, chromatogram A is a chromatogram of a hot water extract of a natural stone mushroom fruiting body re-extracted with ethyl acetate, and B is a chromatogram of an ethyl acetate extract of the extract of the present invention. These chromatograms show that the C. chinensis extract of the present invention contains components that are very similar to those of the natural C. chinensis fruiting body. The Manga extract obtained in this way has almost no toxicity and shows no side effects even when used continuously.
Either as it is or in combination with a food-acceptable carrier according to a conventional method, it can be made into health foods in the form of tablets, pills, granules, capsules, drinks, candy, chocolate, bread, kutsky, etc. be able to. Although it depends on the form of the health food, the L. chinensis extract of the present invention is usually blended in such a way that the daily extract intake (as a powdered extract) is 0.1 to 10 mg/Kg body weight. When used continuously, the health food containing the Cinnamon mushroom extract of the present invention exhibits excellent effects such as nourishment, tonicity, brain health, anti-inflammatory, and diuretic effects. Next, the present invention will be explained in more detail with reference to Reference Examples and Examples. Reference Example 1 39 g of potato dextrose agar medium (manufactured by Difco) was dispersed and dissolved in 1000 ml of water, dispensed into 100 ml flasks in 20 ml portions, autoclaved at 120°C for 30 minutes, and then cooled to prepare a PDA medium. . This was inoculated with 1 platinum loop of C. chinensis seed mycelium (obtained from Kawamura Shiki Shiitake Research Co., Ltd.), and cultured stationary in the dark at 25° C. for 14 days to obtain a seed culture of C. chinensis mycelium. On the other hand, dissolve and dissolve glucose and wheat germ in sterilized water in the proportions shown in Table 1 below, dispense 150 ml each into 500 ml Erlenmeyer flasks, and cap with cotton plugs.
After covering with aluminum foil, it was autoclaved at 121°C for 30 minutes, and then cooled to room temperature to prepare a liquid medium. Three platinum loops of the above seed culture were inoculated into each liquid medium under aseptic conditions, and cultured for 2 weeks in a shaking incubator (220 rpm) in the dark at 25°C. After the culture was completed, the culture solution was centrifuged at 10,000 rpm for 10 minutes, the supernatant was removed, and an appropriate amount of ethanol was added to the precipitate, followed by vigorous shaking. This was filtered by suction on a Buchner funnel, washed with ethanol, thoroughly filtered, and weighed to determine the yield of mycelium. The results are shown in Table 1. Furthermore, in Table 1,
As a control, the results are also shown when the same method was used to culture C. lucidum mycelium using potato dextrose broth as a liquid medium.

[Table] As is clear from the results, when the liquid medium of the present invention is used, a very high mycelium yield can be obtained.
In particular, the wheat germ:glucose ratio is 1:1 to 8.
The yield will be higher. Example 1 Using a jar fermenter, the cultivation of C. chinensis mycelium was carried out as follows. A liquid medium was prepared by mixing 100 g of glucose and 25 g of wheat germ with boiled and cooled tap water in a jar of a jar fermenter (10 volumes), then sealing the jar and autoclaving at 121°C for 30 minutes. did. The jar was attached to a jar fermentor, and 150 ml of a seed mycelial culture, which had been cultured with the same medium at 25° C. for 14 days with shaking, was inoculated under aseptic conditions.
The cells were cultured at 28° C. for 14 days while stirring at 300 rpm and aerating air at a flow rate of 3.0/min. The resulting culture was passed through gauze to obtain 0.84 Kg (wet weight) of mycelium and 3.5 kg of culture solution. Water 2 was added to the separated mycelia, and extraction was carried out by heating at 90 to 100°C for 3 hours, and the extract was concentrated to dryness at 40°C under reduced pressure to obtain 25 g of a brown amorphous powdery L. chinensis extract. In addition, the separated culture solution was concentrated to dryness at 40°C under reduced pressure, and a brown amorphous powdered Stone Manga extract was prepared.
Obtained 55g. No differences were observed between these extracts in gas chromatography, high performance liquid chromatography, thin layer chromatography, paper chromatography, ultraviolet absorption spectroscopy, and various qualitative reactions. Example 2 A health food in the form of tablets was obtained according to the conventional method using the following formulation. Ingredients Quantity Stone mushroom extract (Example 1) 5g Purified water 80ml Lactose 795g 3% hydroxypropylcellulose-lactose
600 g Crystalline cellulose 400 g Potato starch 160 g Talc stearate 40 g From this formulation, 20,000 tablets containing 0.25 mg of Stone Manga extract were obtained. Example 3 A health food in the form of a pill was obtained according to the conventional method using the following formulation. Ingredients Quantity Stone mushroom extract (Example 1) 0.05g Glucose 2.5g Starch 1.5g Gum Arabic 0.8g 100 pills were obtained from this recipe. Example 4 A health food in the form of granules was obtained according to the conventional method using the following formulation. Ingredients Quantity Stone mushroom extract (Example 1) 300g Lactose 2000g Starch 670g Gelatin 30g

[Brief explanation of the drawing]

FIGS. 1 to 3 are a gas chromatogram, a thin layer chromatogram, and a high performance liquid chromatogram, respectively, of the Bamboo shoots extract of the present invention.

Claims (1)

[Claims] 1. Glucose 0.2-10W/V% and wheat germ
Water, alcohol, chloroform, ether, ethyl acetate, benzene, a culture obtained by cultivating Cinderella mycelium in a liquid medium containing 0.2 to 2 W/V% as an essential component, or mycelium separated therefrom;
An extract with a solvent selected from the group consisting of hexane and a mixed solvent thereof or a concentrate of a culture solution from which mycelium has been separated, and is suitable for ninhydrin reaction, α-naphthol reaction, Fehling reaction, phenylhydrazine reagent reaction, anthrone. This is a Cinnamon extract that is positive in all of the sulfuric acid reagent reaction, Biuretz reaction, and ferric chloride reagent reaction, and false positive in the Elson-Morgan reaction. 2. The extract according to item 1 above, wherein the weight ratio of wheat germ to glucose in the medium is 1:1 to 8. 3. The above-mentioned first substance is a solvent extract of isolated mycelia.
Extract of section or section 2. 4. The extract of item 1 or 2 above, which is a concentrate of a culture solution from which mycelium has been separated. 5 Glucose 0.2-10W/V% and wheat germ
Cultivate the Bamboo shoots mycelium in a liquid medium containing 0.2 to 2 W/V% as an essential component, and add the resulting culture or separated mycelia to water, alcohol, chloroform, ether, ethyl acetate, benzene, hexane, and the like. 1. A method for producing a pine mushroom extract, which comprises extracting with a solvent selected from the group consisting of mixed solvents, or concentrating a culture solution from which mycelium has been separated. 6. The method according to item 5 above, wherein the weight ratio of wheat germ to glycose in the medium is 1:1 to 8. 7. The production method of item 5 or 6 above, in which the separated mycelium is extracted with a solvent. 8. The fifth step of concentrating the culture solution from which the mycelium has been separated.
The manufacturing method of Section 6 or Section 6. 9 Glucose 0.2-10W/V% and wheat germ
Water, alcohol, chloroform, ether, ethyl acetate, benzene, a culture obtained by cultivating Cinderella mycelium in a liquid medium containing 0.2 to 2 W/V% as an essential component, or mycelium separated therefrom;
An extract with a solvent selected from the group consisting of hexane and a mixed solvent thereof or a concentrate of a culture solution from which mycelium has been separated, and is suitable for ninhydrin reaction, α-naphthol reaction, Fehling reaction, phenylhydrazine reagent reaction, anthrone. A health food containing, as an active ingredient, a mantis mushroom extract that is positive in all of the sulfuric acid reagent reaction, biuret reaction, and ferric chloride reagent reaction, and false positive in the Elson-Morgan reaction. 10. The health food of item 9 above, wherein the extract is a solvent extract of isolated mycelia. 11. The health food of item 9 above, wherein the extract is a concentrate of a culture solution from which mycelium has been separated.