KR20030026140A - Mycellium production method of Cordyceps militaris, Inonotus obliquus and Phellinus linteus mushroom by use germinated grains and edible filber - Google Patents

Abstract

The present invention is to produce the mycelium ( Phellinus linteus ), Cordyceps spp., Chaga ( Inonotus obliquus ) germinated brown rice and ergot as the main medium material (培 地 材料) Specifically, the method is steamed, germinated brown rice, malt, or moisture-controlled rice bran or varicose bran, and then enters a 850 ml mushroom-growing spawn bottle, and is 121 ° C., 1.2 kg / cm 3. Sterilize in a high-pressure steam sterilizer of about 30 to 90 minutes to cool. And the temperature inside the medium is cooled to room temperature, and then inoculated, inoculated, incubated in a liquid state, 23 to 28 ℃, humidity is characterized in that the culture for 15 to 90 days in a culture room of about 55 to 80% to obtain a mycelium. The obtained mycelium is used as a raw material for beverages or food additives, pharmaceutical raw materials and noodles.

Description

Mycelium production method of Cordyceps militaris, Inonotus obliquus and Phellinus linteus mushroom by use germinated grains and edible filber}

The present invention is a germinated rice medium and dietary fiber rich in cordyceps spp., Phellinus lllinteus , Chaga ( Inonotus obliquus ) germinated brown rice and germinated buckwheat (agar), rice bran, barley bran A method of producing mushroom mycelium by using the rice bran and powder of wheat bran as a medium material, and obtaining functional food additives or pharmaceutical raw materials and raw materials for manufacturing noodles using the mycelium. .

Mushroom fungi are wood-rotting fungi or parasit, plant pathogens, and symbiotic bacteria. By breaking down and growing insects and insects or growing with living trees, they form fruit-bearing mushrooms. In order to artificially cultivate mushrooms that have survived in the natural ecosystem, agricultural or forest by-products such as wood or sawdust and rice straw are used [Ref. (1) G. N. 1988. Plant pathology. Academic press, INC .; (2) Jianzhe, Y., Xiaoloan, M., Quiming, M., Yichen, Z. and Huaan, W. 1989. Icons of meidcinal fungi from China. Science Press. China. (3) Seong Jae-mo et al., Mushroom Science, Teacher's History].

In addition, Cordyceps spp. And Phellinus linteus mushrooms and Chaga mushrooms ( Inonotus obliquus ) have various anti-cancer effects, hypoglycemic effects, blood pressure control effects, immune boosting effects, whitening effects, and nourishing strong effects. Functional [see (4) Kenneth Jones, 1997. Cordyceps Tonic Food of Ancient China, Sylvan Press. (5) Paul Stamets. October 2000. Growing Gourmet and Medicinal Mushrooms. Ten Speed Press. (6) Joan A. Friedrich, William H. Lee. Medicinal Benefits of Mushrooms: Healing for More than 20 Centuries--Their Effects on Cancer, Diabetes, Heart Disease and More, Keats Publishing, Inc. (7) Beth M. Ley. Medicinal Mushrooms for Immune Enhancement: Argaricus Blazei Murill. BL Publications. Reference (8) 前 田 辛 子 et al. 1974, 21: 425-427 (9) Ikekawa J. et al Antitumor action of some besidiomycetes esp . Phellinus linteus. Gann . 59: 155-157, (10) Chung KS et al Effect of Kp. an antitumor protein-polysaccharide from mycellium culture of Phellinus linteus on the humral immune response of tumor-bearing ICR mice to sheep fed blood cell. Arch. of Phar. Res. 16: 336-338].

Such mushrooms are used for edible or medicinal mushrooms or fungi, which are fruiting bodies. However, the use of mushrooms is difficult to overcome seasonal limitations, and the supply is limited. Due to abnormal climate phenomenon, it is becoming more difficult to collect mushrooms. In addition, the artificial production of mushrooms is also difficult to artificially control the environment, expensive production facilities and equipment is required, and the production is also not enough, and is not widely used. Therefore, it is absolutely impossible to secure the raw materials required for the production of processed mushroom products with added functionality.

Therefore, the research on the mass culture method for the production of mycelium (mycelium) that can replace the fruit body (子實体) of the mushroom bacteria has been actively conducted recently.

In the case of Corius sp., Which is a fungus fungi, the method of improving mycelial yield through liquid culture using glucose and yeast extraction medium is disclosed in Korean Patent Publication No. 10-1983-000003 Posted in However, this method differs from the case where the germinated brown rice of the present invention is used as a support for culture because it is intended to obtain mycelium and fruiting body only through liquid culture. In addition, Korean Patent Publication No. 10-1955-007860 discloses that an anticancer / immune-active substance was prepared from a mycelium obtained by liquid culture of Phellus linteus bacteria with glucose, corn slip liquor, and yeast extract as a medium material. have. However, this method is also not appropriate because it does not use a solid medium and is not a raw material production for noodles.

Published in Korean Patent Application Publication No. 10-1996-013174 for the use of S. mushrooms, cultivation of sawdust, rice bran, and water as a main medium material in a plastic bag, and then recovering and using the mycelial film formed on the outer surface of the medium. It is. In addition, Korean Patent Laid-Open Publication No. 10-1996-009897 discloses a method of obtaining mycelium cultured by phagocytosis using Korean ginseng as a culture medium and extracting an extract from the product. However, the present invention does not use materials such as sawdust, rice bran and Korean ginseng as a medium material, but uses grains that are easy to use in manufacturing noodles, that is, germinated brown rice, germinated wheat, malt, and malt sprouts as a medium material. Particularly, in the present invention, the grains are germinated without being used as they are, so that the useful enzymes and polysaccharides produced during the germination of grains can be used as a culture medium for mycelial culture.

As such, the production of mycelia using grain media has been mainly used for situation mushrooms. As known from Korean Patent Publication Nos. 10-1998-044151, 10-1997-062033, and 10-1999-085245, edible grains are used as it is to use only nutrients to help moisture and mycelial growth. It was intended to be used as a raw material such as powder, tea, beverages. The present invention is to form a combination of adenosine (Adenosine) component of Cordyceps sinensis and flavonoids that are abundant in buckwheat germinated malt sprouts (Rutin) natural products (Ref. (11) Ikemoto, T. et al ., 1991, "Physiologically active compounds in the Extracts from Tochukaso and cultured mycellia of Cordyceps and Isaria ." Yakugaku Zassbi 111 (9): 504-509, (12) Melzig, MF, 1996, Inhibition of Adenosine Deaminase activity of aortic endothelial cells by sellected Flavonoids, Plant medica; 62: 20-21] and carbohydrates such as Glucronic acid, Glucose, and Galactose in carbohydrates or dietary fiber in germinated grains It breaks down the homopolysaccharide that forms the sugar-type peptide, and combines it with the heterozygous Xylose peptide and the hexasaccharide arabinose, such as arabixoylan. It was invented as a result of researches that abundance of hetero peptide polysaccharids and glycoproteins (Protein binded polysaccharide) is present.

An object of the present invention is to use agitated as a medium to make a combination with adenosine contained in Cordyceps mycelia of functional natural products and Rutin, a kind of flavonoids abundant in buckwheat sprouts. By producing mycelium, it improves its functionality and cultivates dietary fiber, which is abundant in the germination of germinated brown rice, malt, rice bran, and pulverulent mushrooms, and cultivates cordyceps, green mushroom, and chaga among mushrooms. Produces a mycelium rich in hetero peptide-type polysaccharids that form a bond between hexasaccharides and pentose sugars, which are useful for anti-cancer activity or immunity enhancement and diabetes recovery by decomposing polysaccharides in the form of hexasaccharides homo peptides. It is.

Accordingly, another object of the present invention is to provide a raw material for supplying functional food additives or pharmaceutical preparations and for processing noodles.

1 is a block diagram of the germination process of brown rice and buckwheat.

Figure 2 is a schematic view of the water control germination of brown rice, malt production process.

Figure 3 is a mycelium production method using the germinated grains by the method of the present invention.

Figure 4 is a mycelium production method using rice bran, buckwheat flour by the method of the present invention.

In order to achieve the object of the present invention, Cordyceps spp., Phellinus linteus , Chaga ( Inonotuis obliquus ) bacteria to obtain a mycelium as a raw material rich in germination grains or dietary fiber .

Specifically, 2305CM strain of Cordyceps sinensis strain, PHK41L strain of S. mushroom strain, IN014 strain strain of chaga mushroom strain were cultured on an agar plate in a petri dish to proliferate progenitor. At this time, Cordyceps sinensis, Sichuan mushroom and Chaga mushroom are YMA medium (dextrose 2% (W / V), malt extract 0.3% (W / V), yeast extract 0.3% (W / V), peptone 0.5% (W / V), agar 1.8% (W / V) was incubated for 7 days and used as a progenitor.

Subsequently, the cultured progenitors were propagated through liquid aeration to prepare an inoculator. The inoculum was used by homogenizing agar plate cultured with a homogenizer at 5000-10,000 rpm for 2 to 5 minutes. The reason for the homogenization process by grinding the original bacteria is as follows. First, it is possible to increase the uniformity of the inoculum, and secondly, the clogging of the liquid outlet in the liquid inoculation process due to the agglomeration of the aged hyphae, which is commonly generated in the liquid aeration culture process. Can be excluded. The inoculum propagation medium was heat resistant to 18 liters with glucose 2.5% (W / V), roasted soy flour 0.25% (W / V), peptone 0.5% (W / W), and yeast extract 0.3% (W / V). A plastic bottle was used, and a device designed to prevent contamination of bacteria by attaching a filter of 0.25 μm to the air inlet and the exhaust port for aeration, and for 4 days in the case of Cordyceps sinensis, 7 to 10 for mushrooms and chaga Inoculation was prepared by incubating daily.

After immersion of germinated brown rice, malt, bran, and rice bran, the water is drained until no water flows, and then it is drained for 10-20 minutes at a pressure of 121 ° C. and 1.2 kg / cm 3, preferably about 15 minutes. Steam process The reason for this steaming process is as follows: First, when culturing mycelium using grain medium, it is possible to maintain the gap between the top and bottom of the medium filled inside the culture bottle so that the bacteria, ie bacteria To reduce the defective rate due to the contamination, and secondly, it was carried out to omit the water control step that is necessarily passed in the pre-hospital step, and thirdly to increase the mycelial density of the produced mycelium. On the other hand, by performing steaming in this way, it was possible to continuously maintain mycelial activity, which is presumably because the air permeability is increased by increasing the porosity in the medium.

The medium after the steaming process is put into a 850 ml heat-resistant plastic container and subjected to autoclaving for 30 to 120 minutes under the pressure of 1.2 kg / cm 3 at 121 ° C. using a high pressure steam sterilizer. Inoculation was performed aseptically. After the inoculation process, Cordyceps sinensis is 20 ~ 23 ℃, situation mushrooms and chaga mushrooms are incubated for 15 ~ 75 days in a constant temperature room at 25 ~ 28 ℃. Harvested by cutting. And it is achieved by drying for 12 to 38 hours in a dryer at 45 ~ 65 ℃ for drying the harvested mycelium.

Detailed media composition and mycelium production method of the present invention will be described the process through each experimental example.

[Experimental Example Ⅰ] Production of Cordyceps Sinensis, Situary Mushroom and Chaga Mushroom Mycelium Using Germinated Brown Rice

Course 1: Probiotics Culture

Sichuan mushroom PHK41L, Cordyceps sinensis 2305CM, Chaga mushroom IN014 strain, YMA medium (glucose 2% (W / V), malt extract 0.3% (W / V), yeast extract 0.3% (W / V), peptone 0.5% ( W / V), agar 1.8% (W / V), sterile distilled water 95.5% (W / V) medium was sterilized for 15 minutes at 121 ℃, 1.2kg / cm3 pressure and plated on a Petri dish for 7 days It was cultured and used as a progenitor.

Process 2: homogenization of progeny

100 ml of sterile distilled water was added to a 250 ml grinding cup using a NEISI-25 homogenizer, and the three petri dishes were put in flame sterilized platinum paste using a NEISI-25 homogenizer and the lid of the cup was covered. After setting, the homogenization was performed for 3 minutes at 7500 rpm. All work in this process was carried out in aseptic conditions to minimize contamination of various bacteria.

Course 3: liquid aeration culture

18 liters of homogenized microbial solution is sealed with a silicone stopper using a heat-resistant plastic bottle of mineral water, and an air inlet, a liquid outlet, and an air inlet are installed, and a 0.25 μm pore filter is installed at each of the air inlet and exhaust to prevent invasion of various germs. The top cap of the designed bottle was opened and poured to inoculate. After inoculation, the aeration culture bottle is placed in a culture room at 23 ° C to 25 ° C, injects about 2.0 vvm of air with an oil lees type air pump to inflate bubbles within the liquid medium, and incubates for 5 days. Used.

Course 4: Preparation of Germinated Grains

60 kg by weight of brown rice was submerged in 100 liters of clean water at 18 to 20 ° C. for about 5 hours, and the dehydrated brown rice prepared in advance for germination was drained until dehydration did not occur. And the board was placed in the bottom of the thermostat at 28 ℃ to receive water, and on the head was installed a mesh of 0.05 mesh. At this time, the support rod was installed in the middle to withstand the load of the grain (see Fig. 2). Evenly spread the brown rice extracted after submersion above about 10cm and pour water in one tank every 6-12 hours and sprinkle it on the top grain while turning the grain upside down for 4-5 days to make the rice germinate. do. Germinated grains were taken out of the thermostat when the germinated shoots had a length of about 0.3 to 0.5 cm and used as a medium for producing mycelia of the invention.

Course 5: steaming

After the germination process in step 4, the medium material is subjected to a steaming process for about 15 minutes at 121 ° C and a pressure of 1.2 kg / cm 3.

Course 6: Admission

The steamed medium was cooled in a cooling chamber, and 300 g parts by weight were placed in a 850 ml seed culture bottle.

Course 7: Sterilization

The medium prepared as described above was subjected to high pressure steam sterilization for 35 minutes after reaching 121 ° C and 1.2 kg / cm 3 pressure.

Course 8: Cooling

The sterilized medium is forced to cool until it reaches room temperature in the cooling chamber.

Course 9: Inoculation

After the cooling medium was transferred to the free phase, 25 ml of the liquid culture inoculator prepared in Step 3 was inoculated using an automatic inoculator.

Course 10: Cultivation

The culture was carried out in a constant temperature culture room at 20 to 28 ° C. for 15 to 75 days.

Course 11: Harvest

Mycelium cultured in step 10 was taken out of the culture bottle and harvested.

Course 12: Drying

The harvested medium is dried in a hot air dryer at 45-65 ° C. for 68 hours.

[Experimental Example Ⅱ] Production of Cordyceps Sinensis, Situary Mushroom and Chaga Mushroom Mycelium

Step 1-3: same as Experimental Example I

Course 4: Manufacture of Stirrups

The buckwheat prepared in advance for germination was allowed to immerse 60 kg by weight of buckwheat with respect to 100 liters of clean water at 18 to 20 ° C. for about 3 hours, and was drained out until dehydration was performed. And the board was placed in the bottom of the thermostat at 28 ℃ to receive water, and on the head was installed a mesh of 0.05 mesh. At this time, the support rod was installed in the middle to withstand the load of the grain (see Fig. 2). After immersing the buckwheat collected after the submersion flatly about 10cm flat, and then put water in the bottom tank every 6-12 hours, sprinkle water on the top grain and turn the grain upside down for 4 to 5 days do. Germinated grains were taken out of the thermostat when the germinated shoots had a length of about 0.3 to 0.5 cm and used as a medium for producing mycelia of the invention.

Course 5: steaming

After the germination process in step 4, the medium material is subjected to a steaming process for about 15 minutes at 121 ° C and a pressure of 1.2 kg / cm 3.

Course 6: Admission

The steamed medium was cooled in a cooling chamber, and 300 g parts by weight were placed in a 850 ml seed culture bottle.

Course 7: Sterilization

The medium prepared as described above was subjected to high pressure steam sterilization for 35 minutes after reaching 121 ° C and 1.2 kg / cm 3 pressure.

Course 8: Cooling

The sterilized medium is forced to cool until it reaches room temperature in the cooling chamber.

Course 9: Inoculation

After the cooling medium was transferred to the free phase, 25 ml of the liquid culture inoculator prepared in Step 3 was inoculated by using an automatic inoculator.

Course 10: Culture

The culture was carried out in a constant temperature culture room at 20 to 28 ° C. for 15 to 75 days.

Course 11: Harvest

Mycelium cultured in step 10 was taken out of the culture bottle and harvested.

Course 12: drying

The harvested medium is dried in a hot air dryer at 45-65 ° C. for 68 hours.

[Experimental Example Ⅲ] Production of Cordyceps Sinensis, Situary Mushroom and Chaga Mushroom Mycelium

Step 1-3: same as Experimental Example I

Course 4: Controlling Moisture

Mix 80 liters of rice bran (rice bran) with 35 liters of drinking water and knead it to adjust the amount of water to ooze out slightly when hand-held.

A schematic diagram of the moisture control process is posted in FIG. 2.

Course 5: Admission

300 g parts by weight of each of the 850 ml spawn bottles were performed.

Course 6: Sterilization

The medium prepared as described above was subjected to high pressure steam sterilization for 35 minutes after reaching 121 ° C and 1.2 kg / cm 3 pressure.

Course 7: Cooling

The sterilized medium is forced to cool until it reaches room temperature in the cooling chamber.

Course 8: Inoculation

After the cooling medium was transferred to the free phase, 25 ml of the liquid culture inoculator prepared in Step 3 was inoculated using an automatic inoculator.

Course 9: Cultivation

The culture was carried out in a constant temperature culture room at 20 to 28 ° C. for 15 to 75 days.

Course 10: Harvesting

Mycelium cultured in step 9 was taken out of the culture bottle and harvested.

Course 11: Drying

The harvested medium is dried in a hot air dryer at 45-65 ° C. for 68 hours.

Experimental Example IV Production of Cordyceps Sinensis, Situary Mushroom, and Chaga Mushroom Mycelium

Step 1-3: same as Experimental Example I

Course 4: Controlling Moisture

Mixing 35 liters of drinking water with 80 kg by weight of arc flour (barley bran or wheat bran) and kneading was adjusted so that the water content in the rice bran slightly squeezed out by hand.

Course 5: Admission

The steamed medium was cooled in a cooling chamber, and 300 g parts by weight were placed in a 850 ml seed culture bottle.

Course 6: Sterilization

The medium prepared as described above was subjected to high pressure steam sterilization for 35 minutes after reaching 121 ° C and 1.2 kg / cm 3 pressure.

Course 7: Cooling

The sterilized medium is forced to cool until it reaches room temperature in the cooling chamber.

Course 8: Inoculation

After the cooling medium was transferred to the free phase, 25 ml of the liquid culture inoculator prepared in Step 3 was inoculated using an automatic inoculator.

Course 9: Cultivation

The culture was carried out in a constant temperature culture room at 20 to 28 ° C. for 15 to 75 days.

Course 10: Harvest

Mycelium cultured in step 9 was taken out of the culture bottle and harvested.

Course 11: Drying

The harvested medium is dried in a hot air dryer at 45-65 ° C. for 68 hours.

Germinated grains containing 2 ~ 55% (V / W) containing the mycelium obtained in the above in a boiling water extractor with drinking water and extracted after cooling for 1 to 8 hours while maintaining the pressure of 1.2 ~ 2.5kg / ㎠ Mushroom drink was prepared by using it as it is or by diluting with drinking water or various food additives at an appropriate ratio. And the mycelium using germinated grains or rice bran and wheat flour as a medium is provided as a raw material for functional polysaccharide extraction.

As described above, the method of producing a food additive and a functional polysaccharide extract raw material containing the mycelium of the present invention uses the grains of rice bran and wheat rich in germinated grains and dietary fiber as compared to the case of using grains as it is. Faster mycelial growth can shorten the production period, reduce invasion of various germs, and increase its functionality, making it easy to use as a dietary ingredient for improving noodles, porridge, rice, beverages, tea or constitution. In addition, the mycelia produced by the present invention have the effect of increasing the yield of functional polysaccharides by being degraded by the mushroom bacteria dietary fiber that is abundantly present in germinated grains and rice bran, whistle.

Claims (9)

After germination of the germinated grains, the bacteria were grown in a mushroom seed strain, sterilized, and then cultured in a liquid state, Phellinus linteus , Cordyceps spp., Or Chaga ( Inonotus obliquus ) were cultured. A method for producing mushroom mycelia, inoculating and culturing. The method for producing mushroom mycelium according to claim 1, wherein the germinated grains are germinated brown rice or wort sprouts. The method of producing mushroom mycelium according to claim 1, wherein the steaming is carried out at 121 DEG C and a pressure of 1.2 kg / cm < 3 > for 10-20 minutes. The method for producing mushroom mycelium according to claim 1, wherein the sterilization is carried out in a high pressure steam sterilizer at about 121 ° C and about 1.2 kg / cm 3 for 30 to 120 minutes. The method for producing mushroom mycelium according to claim 1, wherein the culture after inoculation of the bacteria is carried out in a culture chamber at 23 to 28 DEG C and a humidity of 55 to 80% for 15 to 90 days. Moisture and steaming of rice bran or water are then steamed, inoculated into mushroom cultivation seedlings , sterilized, and cultured in liquid state, Phellinus linteus , Cordyceps spp., Or chaga . A method for producing mushroom mycelium, characterized by inoculating mushrooms ( Inonotus obliquus ), incubating for 15 to 90 days in a culture room at 23 to 28 ° C. and a humidity of 55 to 80%. 7. The method for producing mushroom mycelium according to claim 6, wherein the steaming is carried out at 121 DEG C and a pressure of 1.2 kg / cm < 3 > for 10-20 minutes. 7. The method for producing mushroom mycelium according to claim 6, wherein sterilization is carried out for 30 to 120 minutes in a high pressure steam sterilizer at about 121 deg. The method of producing mushroom mycelium according to claim 6, wherein the culture after inoculation of the bacteria is carried out for 15 to 90 days in a culture chamber at 23 to 28 DEG C and a humidity of 55 to 80%.