JP5731082B2 - An ergothioneine production method and production apparatus using mushroom basidiomycetes using a submerged culture method. - Google Patents

Description

  The present invention relates to a method for producing a useful substance using basidiomycetes or ascomycetes, and in particular, after filtering off mycelia of basidiomycetes or ascomycetes that have been submerged in a liquid medium, the mycelium is adjusted to moisture and mycelia And producing a useful substance at low cost by increasing the production of a useful substance in the mycelium by applying a stress such as low-temperature resting to the mycelium and storing the mycelium in a living environment. . In the present invention, a useful substance means an antioxidant substance such as ergothionein, an antibiotic substance such as penicillin, a substance that suppresses hematopoietic damage due to radiation, a basidiomycete means a mycelium of mushrooms, and mushrooms include enokitake mushrooms and tamagotake mushrooms. , Eringi, agitake, tokihirohiratake, ganoderma and the like. The mycelium of ascomycetes means mycelium such as trif and blue mold.

  Regarding the method for producing ergothioneine, Patent Document 1 discloses a method for producing ergothioneine by extracting and purifying ergothioneine from the mycelium obtained by culturing mycelia of mushrooms. Patent Document 2 discloses a method for cultivating mycelia of Tamogitake in a medium containing methionine and extracting and producing ergothioneine from the produced mycelium, and here, mushrooms are produced using a liquid medium. After culturing, the mycelium is filtered, freeze-dried, pulverized to extract the target useful substance, and a method for producing the mycelium by filtering off the useful substance ergothioneine is disclosed. Yes. In these prior documents, mushrooms are cultured using a liquid medium, and then the mycelium is taken out, and the taken out mycelia are dried to extract ergothioneine. Ergothioneine is known to have an antioxidant effect and an anti-aging effect, and is used in cosmetics, foods, pharmaceuticals, feeds and the like.

  In such a conventional technique, when the mycelia cultured in liquid culture are filtered off, the culture medium is lost, so that it is removed from the deep culture state, and the production activity of useful substances (ergothioneine) by the mycelium is stopped. Note that the mycelium is killed when the mycelium is filtered and lyophilized. Therefore, the production of ergothioneine, a useful substance, by mycelium has been limited to the period of liquid culture. On the other hand, the liquid culture requires a lot of cost, and thus there is a problem that the production cost of the intended useful material becomes expensive. Therefore, a method of providing useful substances at low cost by producing mycelium is desired.

JP 2009-159920 A

JP 2012-105618 A

  An object of the present invention is to provide an inexpensive mycelium-derived useful substance by solving the above-mentioned problems, and after filtering the mycelium of basidiomycetes or ascomycetes cultured deeply in a liquid medium, the mycelium A method for producing a useful substance by increasing the production of a useful substance by adjusting moisture in the body to produce a mycelium, storing the mycelium in a living environment, and applying a stress such as low-temperature rest to the mycelium It is to provide a manufacturing apparatus.

  In the present invention, useful substances are antioxidant substances such as ergothioneine, antibiotics such as penicillin, substances that suppress hematopoietic damage due to radiation, mycelia of basidiomycetes are mycelium of mushroom fungi, and mushrooms are enokitake mushroom and tamagotake mushroom. , Reishi, cordyceps and so on. Examples of the mushroom fungus used for the production of ergothionein include Tamogitake fungus, Agitake fungus, Tokihirohiratake fungus, Eringi fungus, etc. in addition to Enokitake fungus. The mycelium of the ascomycete is a mycelium such as Trifungus or Blue mold.

The invention according to claim 1 is a method for producing ergothioneine by deeply culturing mycelia of mushroom basidiomycetes in a liquid medium and extracting and purifying ergothioneine from the produced mycelium, wherein a submerged culture step (step) The mycelium cultured in S1) is removed from the deep culture state, a filtration treatment is performed to reduce water from the culture solution containing the mycelium and the liquid medium in the culture tank, and the mycelium to which the liquid medium is attached is removed. Filtration step (step S2) for sorting, and producing a mycelium of the filtered mycelium, and adjusting the water content to make the mycelium viable in the air environment adjustment process (step S3), and the mycelia mass water content has been adjusted and stored under survival environment, static置刺after thermal stimulation or electrical stimulation applied under static置刺intense and low temperature environment to the hypha masses Wherein the ergothioneine by applying a temperature stimulation is ergothioneine manufacturing method according basidiomycete mushroom using a stress application step of producing increased amounts of mycelia mass (step S4), and submerged culture method consisting and in a low temperature environment And

In the present invention, Flammulina fungus mycelium, Pleurotus cornucopiae fungus, Reishibakin, can be any mycelium selected from Cordyceps bacteria, in the present invention, eryngii bacteria, Agitake bacteria as mycelium, collected by The mycelium of the oyster mushroom can be used. Moreover, the mycelium of Trifoccus which is an ascomycete can also be utilized.

In the present invention, the stress applied to the mycelial mass, static置刺stimulation, thermal stimulation, electrical stimulation, can be either stimulated or stimulated with a combination of these two or more selected from stimulation by gas, the In the present invention, the stress applied to the mycelium can be a stimulus by a change in humidity, a stimulus by a change in pH environment, or a stimulus by a drug.

The invention described in claim 2 includes a culture tank 13 for accommodating a liquid medium 11 therein and performing deep culture of mycelia of mushroom basidiomycetes 12, an airtight hatch 14 attached to the upper part of the culture tank, and a culture tank. A stirring mechanism 15 that stirs the liquid medium, a sterile air supply mechanism 16 that supplies a predetermined amount of sterile air to the liquid medium, a pH adjustment mechanism 17 that adjusts and maintains the pH of the liquid medium to a predetermined value, and a liquid. A deep portion for deeply culturing mycelia, comprising a temperature adjustment mechanism 18 for adjusting and maintaining the temperature of the culture medium at a predetermined value and an exhaust mechanism 19 for exhausting air supplied from the sterile air supply mechanism to the outside from the culture tank The mycelium cultured by the culture means 10 and the deep culture means is taken out of the culture tank and removed from the deep culture state, and moisture is removed from the culture solution CF containing the mycelium and the liquid medium in the culture tank. A filtration means 20 for separating the filtered culture liquid into a sol or gel-like mycelium HS and a post-filtration culture liquid AFCF, and the filtration means. A mycelium water content adjusting means 30 for generating a mycelium mass HSL of the mycelium and adjusting the water content contained in the mycelium to an optimum water content for survival of the mycelium, and the mycelium water content adjustment Store the mycelium of which moisture content is adjusted by means in a living environment, and leave the stimulation against static stimulation and temperature stimulation under low-temperature environment or static stimulation after applying electrical stimulation to the mycelium and temperature stimulation under low-temperature environment. the given, the stress applying section 40 for generating a ergothioneine bulking hypha masses IIHSL with increased production of ergothioneine, ergothioneine manufacturing instrumentation by basidiomycete mushroom with deep culture method having a And characterized in that. Here, the sol or gel-like mycelium means a state in which the mycelium is contained in the liquid.

  In the present invention, the mycelium of Enokitake, Tamogitake, Ganoderma or Cordyceps fungus can be used as the mycelium, and the useful substance can be ergothioneine. In addition, as stress applied to the mycelium, any one selected from static stimulation, temperature stimulation, electrical stimulation, and gas stimulation, or a combination of two or more of these stimulations can be used.

  The present invention provides a process for producing a mycelium by adjusting the moisture content of the mycelium after the submerged mycelium is filtered, and a process for applying stress by storing the mycelium in a living environment. As a result, the mycelium mass rapidly and in large quantities produces useful substances to cope with the applied stress. Therefore, a large amount of useful substances can be produced in a short period of time using mycelia cultured in the deep part, and the production cost of useful substances can be greatly reduced.

  In the present invention, since the stress application step is provided, the same mycelium as the mycelium that produced the useful substance in the deep culture step is added to produce the useful substance, so that it is the same as the raw material used for the deep culture. Useful materials can be obtained several times from the amount of raw materials, and the raw material costs of useful materials can be greatly reduced.

It is a flowchart which shows the main processes for enforcing the useful substance manufacturing method concerning this invention. It is a figure explaining the structural example and manufacturing process of the useful substance manufacturing apparatus which concern on this invention.

  Hereinafter, embodiments of a useful substance production method and production apparatus using basidiomycetes or ascomycetes using a deep culture method according to the present invention will be described with reference to the accompanying drawings. FIG. 1 is a flow chart showing a procedure for producing a useful substance produced by a mycelium according to the present invention. In the present invention, first, a process of deeply culturing mycelia of basidiomycetes or ascomycetes (hereinafter referred to as “mycelia”) using a liquid medium to proliferate mycelium (step S1), and the culture is completed. After filtering the culture solution containing the mycelium and the liquid medium in the culture tank and separating the mycelium, the water content of the mycelium is adjusted to produce a mycelium mass Mycelium moisture content adjustment step (step S3), and then storing the mycelium in a living environment, applying various stresses to the mycelium to increase production of useful substances, increasing the useful substances The stress application process (step S4) which produces the mycelium which performed is performed. The mycelium filtered in the filtration step (step S2) is discontinued from the deep culture state due to the loss of the culture solution, and the mycelium stops growing. Will be.

  FIG. 2 is a diagram for explaining a configuration example and a manufacturing process of a manufacturing apparatus for carrying out the useful substance manufacturing method according to the present invention. The useful substance manufacturing apparatus (hereinafter referred to as “manufacturing apparatus”) according to the present invention includes a deep culture means 10, a filtering means 20, a mycelium moisture content adjusting means 30, and a stress applying means 40.

  In the deep culture means 10, a culture tank 13 for culturing mycelium is a container for accommodating a liquid medium 11 therein and performing deep culture of mycelium 12. The airtight hatch 14 is attached to the upper part of the culture tank 13 and used for charging raw materials and cleaning the tank. The stirring mechanism 15 is connected to the culture tank and stirs the liquid medium in the culture tank. The sterile air supply mechanism 16 is connected to the culture tank and supplies a predetermined amount of sterile air to the liquid medium. The pH adjusting mechanism 17 is connected to the culture tank and adjusts and maintains the pH value of the liquid medium to a predetermined value. The temperature adjustment mechanism 18 is connected to the culture tank and adjusts and maintains the temperature of the liquid medium to a predetermined value. The exhaust mechanism 19 exhausts the air supplied from the sterile air supply mechanism to the outside from the culture tank.

  The culture solution CF is delivered from the deep culture means 10 and supplied to the filter separation means 20, and is a liquid containing mycelium and a liquid medium that have been deeply cultured inside the culture tank. The filtering means 20 is supplied with the culture solution CF from the culture tank 13 of the deep culture means 10, and the culture solution is aseptically separated into the mycelium HS and the filtered culture solution AFCF to remove the mycelium from the deep culture. Let go. The mycelium HS is a sol or gel mycelium separated and removed from the culture solution CF by the filtering means 20, and the post-filtering culture solution AFCF is the remaining culture solution obtained by removing the mycelium HS from the culture solution CF. is there.

  The mycelium moisture content adjusting means 30 receives the mycelium HS supplied from the filtering means 20 and aseptically adjusts the water content contained in the mycelium HS to the optimum content for survival of the mycelium. Generate a mass HSL. The mycelium lump HSL is a mycelium lump that has been adjusted to an optimal state for the survival of the mycelium and the mycelium HS has been detached from the submerged culture and is alive in the air. The stress applying unit 40 receives the mycelium lump HSL from the mycelium moisture content adjusting unit 30 and applies various stresses ST to the mycelium lump HSL. When the mycelium is subjected to stress by the stress applying means 40, the production activity of the intended useful substance is activated and the useful substance is greatly increased. Thus, the stress applying means 40 generates the useful substance-increasing mycelium IIHSL in which the useful substance is increased.

  Next, the production procedure of the useful substance according to the present invention will be described in detail with reference to FIG. First, the airtight hatch 14 is opened, and the liquid medium 11 is introduced into the culture tank 13. The temperature control mechanism 18 maintains the inside of the tank at a high temperature around 121 ° C. to sterilize the liquid medium inside. When the sterilization is completed, the temperature control mechanism 18 cools the culture tank and adjusts the internal liquid medium to a temperature suitable for inoculation of mycelia to about 5 ° C. to 35 ° C. to maintain the temperature.

  Then, basidiomycetes or ascomycetes are inoculated from the airtight hatch 14 into the liquid medium 11 in the culture tank. The liquid medium stirring mechanism 15 stirs the liquid medium in the culture tank. The sterile air supply mechanism 16 introduces air into the liquid medium in the form of bubbles and performs deep culture. The air introduced into the culture tank 13 is discharged to the outside by the exhaust mechanism 19. The pH adjusting mechanism 17 adjusts the pH value of the liquid medium during the submerged culture period to about pH 3 to pH 9, and maintains the pH value. Thereby, the deep culture process (step S1) is executed. When the deep culture is completed, the mycelium of basidiomycete or ascomycete is elongated in the liquid medium to obtain the culture solution CF.

  The culture solution CF is supplied to the filtration means 20. In the filtration process (step S2), the filtration means 20 uses a filter filtration, a centrifugal filtration, a vacuum filtration or a cyclone filtration in an aseptic environment, for example, an outside air closed environment, and the culture solution CF is separated from the mycelium and cultured. It is separated from the liquid by filtration to produce sol or gel mycelium HS. The mycelium HS that has been filtered out is disengaged from the deep culture state due to the loss of the culture solution and stops the mycelial growth activity.

  The filtering unit 20 supplies the mycelium HS to the mycelium moisture content adjusting unit 30. In the mycelium moisture content adjusting step (step S3), the mycelium moisture content adjusting means 30 adjusts the moisture content of the mycelium HS by drip of moisture by pressing, drying, centrifugal dehydration or dead weight under an aseptic environment. The mycelium HSL is generated by adjusting the hyphae to an optimum value for survival in the air, about 75% to 99%. The mycelium lump HSL has a moisture content that is optimal for survival, and it is placed in an air environment with a temperature of 5 ° C to 35 ° C and a humidity of 50% to 100%, which is optimal for survival. Has continued production activities.

  The mycelium moisture content adjusting means 30 supplies the surviving mycelial mass HSL to the stress applying means 40. In the stress applying step (step S4), the stress applying means 40 applies various stresses (stimulations) ST as described below to the mycelium lump HSL. The mycelium lump HSL is stored for about 2 to 20 days in a low temperature around 6 ° C or a high temperature around 30 ° C, or in an environment where the low and high temperatures are changed (stationary stimulation, temperature stimulation). . An electrical stimulus such as applying a spark discharge by direct current to the mycelium lump HSL for about 2 to 10 seconds is given. The mycelium HSL is stimulated by gas by adding carbon dioxide gas or ozone gas to the air environment at a predetermined concentration for a certain period of time. In addition, the mycelium lump HSL is kept stationary for about 2 to 20 days in a low humidity of about 60% or a high humidity of about 90%, or in an environment where low and high humidity are changed (humidity stimulation). . The mycelium HSL is stored for about 2 to 20 days in an environment where the pH value is maintained around 5 or around 8 or the pH value is changed (stimulation due to a change in pH environment). The mycelium HSL is sprayed with sulfuric acid or caustic soda solution diluted to a predetermined concentration with sterilized water, and left to stand for about 2 to 20 days (stimulation with chemicals). In the present invention, any one stress selected from the above stresses can be employed, but a combination of two or more of these stresses can be employed. The mycelium HSL receives the stress ST and activates the production activity of useful substances in order for the mycelium to cope with the environmental change due to the application of stress. As a result, useful substance-increasing mycelium IIHSL with increased useful substance is produced.

  The intended useful substance can be extracted from the useful substance-enriched mycelium IIHSL, and the resulting extract can be purified by known methods such as centrifugation, crystallization, ultrafiltration, evaporator, activated carbon treatment and the like. When a useful substance such as penicillin is contained in the culture broth CF, the useful substance can be purified from the extract of the culture broth AFCF after filtration and the hyphae IIHSL after increasing the useful substance by a known method. it can. According to the present invention, in addition to the useful substance produced and obtained in the submerged culture, the useful substance produced by greatly increasing the useful substance by applying stress can be obtained.

  In addition to the useful substance produced by the mycelium in the step of culturing deeply and growing the mycelium as described above (step S1), the mycelium is filtered from the culture solution (step S2), By executing the step of adjusting the moisture content to generate a mycelial mass (step S3) and the step of storing the mycelial mass in a living environment and applying stress (step S4), the mycelium is obtained from the culture solution. Rapidly produce a large amount of useful substances to cope with the stress applied under severe conditions where surrounding nutrients are scarce by leaving the deep culture state from the favorable deep culture environment in the atmosphere I think that.

  Using the production apparatus according to the present invention, ergothioneine was produced as follows using enokitake mycelia as mushroom mycelia. As primary culture, enokitake mushroom mycelia were plated on potato dextrose agar (PDA) medium to obtain the original seed fungus. As a secondary culture, the mushroom fungus with a flat plate was removed from the original inoculum obtained in the primary culture to a diameter of 5 mm with a cork borer, sterilized and cooled in an Erlenmeyer flask, and 200 ml (milliliter) of MYPG medium (malt extraction) 2.76 g / l (liter), yeast extract 3.84 g / l, glucose 3.84 g / l, porcine pancreatic digested casein peptone 3.84 g / l), etc. Liquid inoculum was obtained.

  30 g / l soy flour added to water and extracted for 30 minutes, sugar 20 g / l, magnesium sulfate heptahydrate 0.5 g / l, potassium dihydrogen phosphate 0.5 g / l, silicon The culture solution to which 0.5 g / l of antifoam was added was filled into the culture tank from an airtight hatch. The culture solution inside the culture tank was sterilized at a temperature of 121 ° C. for about 20 minutes and then allowed to cool to maintain the temperature of the culture solution at about 20 ° C.

  Then, the liquid inoculum obtained from the secondary culture from the airtight hatch is inoculated into the culture solution in the culture tank, and air is introduced into the liquid medium from the aseptic air supply mechanism in the form of bubbles and deep culture is performed for about two weeks. It was. The air introduced into the culture tank was evacuated, and when the submerged culture was completed, a culture solution in which enokitake mushrooms were elongated in the liquid medium was obtained. Here, the deep culture process (step S1) ends.

  The culture solution obtained in the submerged culture was aseptically filtered in an airtight environment into enokitake mycelium and culture solution using a mesh-like stainless steel filter to obtain sol-like or gel-like enokitake mycelium ( Filtration process step S2). Here, the mycelium is detached from the deep culture state due to the loss of the culture solution. Subsequently, the sol-form or gel-form enokitake mycelium is placed on a stainless steel mesh in a sterile environment, an appropriate weight is placed on the mycelium, and the moisture of the mycelium is dripped by drip of the water by pressing. The content rate was adjusted to 80 to 98% to obtain a mycelium mass (mycelium mass) (mycelium moisture content adjustment step S3). Here, the mycelial mass produced aseptically had the above-mentioned water content optimal for survival, and was stored in an environment of temperature and humidity optimal for survival, so that survival was confirmed.

  Then, the mycelial mass was stored in a low temperature environment of about 6 ° C. for 20 days, so that the mycelial mass was subjected to static stimulation and temperature stimulation (oxidation stress) (stress application step S4). Here, it is considered that the mycelial mass is rapidly put into mass production of the antioxidant ergothioneine by being placed in an oxidative stress environment, and the mycelial mass containing a large amount of ergothioneine (ergothioneine-enriched mycelial mass) I was able to get it. Then, the hyphae lump is dried and pulverized on the first day, the first day, the third day, the sixth day, the ninth day, the twelfth day, and the twentieth day. Ergothioneine was extracted from the resulting product using an organic solvent, and the ergothioneine content of a purified product obtained by purifying the resulting extract by a general method was measured. Table 1 shows the HPLC analysis results showing the relationship between the stationary storage elapsed date of the mycelium and the ergothioneine content of the purified product on that day.

  As shown in Table 1, the content of ergothioneine by adding static stimulation and temperature stimulation (low-temperature static stress) to the enokitake mushroom mycelium reached the maximum value (1.70) on the 12th day. It can be seen that the content is about 6.07 times the content (0.28) of the day (first time) when the stationary storage is started, which is almost equivalent to the production amount of another enokitake mycelium.

  After applying a spark discharge of DC 100 KV for 3 seconds to the enokitake mushroom mycelium obtained in the same manner as in Example 1 (voltage application stress), the mycelial mass was stored in a low temperature environment of about 6 ° C. for 20 days. And, as in Example 1, on the day of starting stationary storage (first departure), the first day, the third day, the sixth day, the ninth day, the twelfth day, and the twentieth day, ergothioneine The ergothioneine content of the extracted and purified product was measured. Table 2 shows the HPLC analysis results showing the relationship between the stationary storage elapsed day of the mycelial mass after voltage application and the ergothioneine content of the purified product on that day.

  As shown in Table 2, the content of ergothioneine by applying low-temperature static stress to the enokitake mushroom mycelium after voltage application reached the maximum value (1.92) on the 9th day, and the day when the static storage was started (first time ) Content of about 6.85 times (0.28). In addition, the content of ergothioneine on the day when the stationary storage is started (first time) substantially corresponds to the content of enokitake mycelium separated from the culture solution (content according to the prior art). This fact is that Example 2 which applied stress by applying voltage to low temperature standing reached the maximum value of ergothioneine content three days earlier than the first example which applied only low temperature standing stress. It can also be seen that the content is increased by 0.22.

  As is clear from the above examples, the mycelium is added to the ergothionein produced in the submerged culture step by performing the mycelium moisture content adjustment step and various stress application steps after performing the filtration step. Rapidly mass-produces ergothioneine, an antioxidant. Moreover, by adding low temperature static stress to the mycelium after voltage application, the increase rate of ergothioneine can be increased and the content can be increased. As mushroom fungi used for the production of ergothioneine, in addition to Enokitake fungi, Tamogitake fungi, Ganoderma fungi, Cordyceps fungi, Agitake fungi, Tokihirohiratake fungi, Eringi fungi and the like may be used. Moreover, you may use a Trifungus etc. as an ascomycete.

  According to the present invention, antioxidants such as ergothioneine that can be used in feeds, cosmetics, medicines, antibiotics, etc. can be produced at low cost by the mushroom mycelium. It can contribute widely in the medical industry and has a wide range of uses such as pharmaceuticals, cosmetics, and health foods.

DESCRIPTION OF SYMBOLS 10 Deep culture | cultivation means 11 Liquid culture medium 12 Basidiomycetes or Ascomycetes 13 Culture tank 14 Airtight hatch 15 Stirring mechanism 16 Aseptic air supply mechanism 17 pH adjustment mechanism 18 Temperature adjustment mechanism 19 Exhaust mechanism 20 Filtration means 30 Mycelium moisture content adjustment means 40 Stress application means CF culture medium HS mycelium AFCF culture medium after filtration HSL mycelial mass ST stress II HSL useful substance increase mycelial mass

Claims (2)

A method for producing ergothioneine by deeply culturing mycelia of mushroom basidiomycetes in a liquid medium, extracting and purifying ergothioneine from the produced mycelium,
The mycelium cultivated in the deep culture step is removed from the deep culture state, filtered to reduce the moisture from the culture solution containing the mycelium and the liquid medium in the culture tank, and the mycelium to which the liquid medium is attached A filtration process for separating
Producing a mycelium of the filtered mycelium, and adjusting the water content to adjust the water content in order to make the mycelium viable in an air environment; and
Store the mycelium of which the moisture content is adjusted in a living environment, and subject the mycelium to a static stimulus in a static environment and a low temperature environment or a static stimulus after applying an electrical stimulus and a thermal stimulus in a low temperature environment. Applying a stress to produce a mycelium of increased ergothioneine by application,
The manufacturing method of the ergothioneine by the mushroom basidiomycetes using the deep culture method characterized by comprising . A culture tank that contains a liquid medium inside and performs deep culture of mycelium of mushroom basidiomycetes , an airtight hatch attached to the top of the culture tank, a stirring mechanism that stirs the liquid medium in the culture tank, and a liquid medium A sterile air supply mechanism for supplying a predetermined amount of sterile air to the liquid, a pH adjusting mechanism for adjusting and maintaining the pH of the liquid medium to a predetermined value, and a temperature adjusting mechanism for adjusting and maintaining the temperature of the liquid medium to a predetermined value An exhaust mechanism for exhausting the air supplied from the sterile air supply mechanism to the outside from the culture tank, and a deep culture means for deeply culturing mycelium,
The mycelium cultured by the deep culture means is removed from the culture tank and removed from the deep culture state, and a filtration treatment is performed to reduce water from the culture solution containing the mycelium and the liquid medium in the culture tank. A filtration means for separating the filtered culture solution into a sol or gel mycelium and a culture solution after filtration;
Generating a mycelium of the mycelium filtered by the filtering means, and adjusting the moisture content contained in the mycelium to an optimal moisture content for survival of the mycelium; and
The mycelium whose water content is adjusted by the mycelia water content adjusting means is stored in a living environment, and the mycelium is subjected to static stimulation and static stimulation after application of temperature stimulus or electrical stimulation in a low temperature environment. and giving thermal stimulation in a low temperature environment, and stress applying means for generating a ergothioneine bulking mycelial mass increased production of ergothioneine,
An apparatus for producing ergothioneine using mushroom basidiomycetes using a submerged culture method.