JP2688497B2 - Multistage solid culture method and device - Google Patents

Description

TECHNICAL FIELD The present invention relates to a solid culture method and an apparatus therefor, which is a multistage solid culture method used for culturing microorganisms for production of useful substances and enzymes. And its device.

(Prior Art) Conventionally, there are a tray method and a ventilation deposition method for solid culture, and the tray method is performed by depositing a medium in a tray and covering it with a lid.
The ventilation deposition method is a method in which a solid medium is forcibly ventilated and culture is performed while controlling the environment such as humidity.

In addition, a multistage shelf culture method in which solid medium is diffused and arranged in multiple stages to supply conditioned air, and a rotary drum double-sided culture method in which the rotating drum doubles as a steaming can and the medium is stirred by rotating the drum Etc.

Each of the above conventional methods has the following drawbacks.

 That is, the water content of the solid medium is not properly adjusted during the culture.

Removal of metabolic heat generated by the growth of microorganisms is not successful.

The supply of oxygen required to promote the growth of microorganisms is insufficient.

Therefore, JP-A 61-128882 has been proposed for the purpose of eliminating these drawbacks and performing efficient production.

In this method, the aeration culture method is used as a skeleton, and the water supply of the solid medium is controlled in relation to the shaft torque of the stirring shaft to carry out large-scale culture.

(Problems to be solved by the invention) In the solid culture method in the prior art, in order to prevent contamination of various bacteria and drying of the medium, etc., it is carried out in a closed chamber to form a simple aseptic chamber, but due to lack of oxygen, A ventilation method is used to prevent a decrease in production.

In order to prevent drying by aeration, aeration, humidity control of air, and water spray supply to a solid medium are carried out, but this is extremely difficult.

In Japanese Patent Laid-Open No. 61-128882, the water supply is controlled in relation to the shaft torque of the stirring shaft, but it is easy to control because it is a monolayer culture.

If multi-stage shelf culture is used for the purpose of large-scale culture, if the aeration water is supplied in common to each stage,
Aeration problems occur, it is difficult to control the water content, and it is a stationary type, which causes problems such as lack of oxygen inside the culture due to adhesion between solid media.

Therefore, when a multi-stage system is adopted for the purpose of large-scale culture, it is necessary to satisfy the conditions that water content can be adjusted and aeration can be performed for the purpose of oxygen supply and metabolic heat removal.

It is an object of the present invention to provide a multistage culture method and device that enable large-scale culture.

(Means for Solving the Problems) The present invention takes the following means in order to achieve the above objects.

That is, as the multi-stage solid culture method of the present invention, the medium trays for storing the solid culture are stacked in multiple stages through a separation frame in a sterile culture chamber to which sterile air is supplied, and the stirring blades in each medium tray are used. While stirring the medium, aseptic air is supplied to each stage from the lower side of the medium tray and discharged from the upper side, and moisture is supplied from each different moisture supply nozzle toward the solid medium in the medium tray of each stage, While setting the environment according to the culturing status of the stage, the means for culturing while controlling the sterile air and water is used.
It is controlled by detecting the strain value of the stirring blade that is generated from the relationship between the stirring blade that stirs the solid medium and the friction coefficient between water and the medium that grows the bacteria.

And as an apparatus for carrying out the above method,
A holding tray for storing solid medium, a holding tray for solid culture, a rotatable stirring blade for stirring the solid medium, and a culture tray with a sterile air inlet on the lower peripheral wall of the holding net, and a sterile wall on the peripheral wall of the frame. The separation frame equipped with an air exhaust port and a water supply nozzle that supplies water to the solid medium is alternately stacked in multiple stages in a sterile culture chamber, and the rotation axis of the stirring blade of each medium tray is the rotation axis of the separation frame. The water supply is controlled by a strain gauge attached to the stirring blade of the culture tray and a signal sent to the water control device via a slip ring. The output signal is output to the supply valve of the water supply nozzle to control the supply valve to be turned on and off.

In addition, the above-mentioned medium tray and the separation frame are stacked in multiple stages, and a guide shaft that fits with each guide piece of the medium tray and the separation frame and guides them vertically to a certain direction and a certain position, and the tray. And a guide rail for moving the separation frame to the guide shaft from the inlet of the aseptic culture chamber, and a transport device for guiding the medium tray and the separation frame along the rail, and the tray and the separation frame are guided by the guide shaft. And a cylinder device for temporarily holding the tray and the separation frame at a constant height.

(Function) The cultivation trays that store the solid cultivations are stacked together with the separation frame in a sterile culture chamber in which aseptic air is alternately supplied, and the culture trays at the bottom of the culture tray are stirred while stirring the culture medium with the stirring blades inside the culture tray. Aseptic air is supplied from the air inlet, exhausted toward the peripheral wall of the sterile culture chamber from the exhaust port of the separation frame mounted on the upper part through the space between the culture mediums, rises, and is exhausted from the exhaust port of the sterile culture chamber ceiling.

For the water supply, the strain value of the strain gauge attached to the stirring blade of the medium tray is detected from the relationship between the water content and the friction of the medium, and as a result, the water is supplied from the water supply nozzle attached to the lower part of the separation plate of the separation frame. It is sprayed under control.

(Example) Hereinafter, an example illustrated in the drawings will be described.

The solid culture method has many inconveniences such as complete sterilization of the solid medium and difficulty in managing the culture conditions, but in particular, in the case of fungal extracellular enzymes, the production amount in bran solid medium is often extremely high. By extracting the solid medium after culturing with water, there is an advantage that a relatively concentrated enzyme solution can be easily obtained.

In addition, solid-state culture is a method generally suitable for enzyme production of mold as seen in the production of takadiastase, and even now, glucoamylase by Rhizopus including Takadiastase, cellulase by Aspergillus niger,
Hemicellulase, pectinase, cellulase by Trichoderma, lactase by Aspergillus oryzae, Muco
The major enzymes of fungi, such as the production of microbial rennet by r, are produced by solid culture.

However, as described above, even if suitable for the production of enzymes by mold, solid culture has poor heat conductivity as one of them, and the amount of heat accumulated due to rapid propagation of microorganisms is smoothly dissipated. Since it does not exist, the temperature of the bran increases and the growth of target microorganisms is suppressed, while promoting the growth of thermophilic bacteria and causing contamination.

If the bran layer is thicker than a certain degree because the air permeability is poor, the inside becomes anaerobic and the growth of the target microorganism is suppressed.

Therefore, it is necessary to culture while controlling the temperature, humidity and aeration.

The raw material for the solid culture medium is wheat bran, which is rarely a mixture of about 20% of rice bran or soybean meal with wheat bran.

Then, a solid medium supplemented with appropriate salts is used, and after inoculation, it is thinly spread on a medium tray.

As described above, the medium tray in which the solid medium is spread thinly is
Together with the separation frame, they are stacked alternately in a sterile culture chamber.

In order to improve aeration, while stirring the solid medium with the stirring blades in the medium tray, sterile air is supplied to each stage from the lower side of the medium tray and discharged from the upper side, and the difference in the amount of water added to the solid medium is caused by the microorganisms. Since it greatly affects not only the growth of but also the enzyme production, water is sprayed from the water supply nozzle.

The amount is controlled by detecting the distortion of the stirring blade.

Then, the culture is performed while controlling the sterile air and water while setting the environment according to the culture condition of each stage.

Although it is difficult to continuously measure changes in temperature and pH during culture, cell growth, consumption of medium raw materials, and changes in components, it is difficult to measure the friction coefficient between the stirring blade and the medium due to water and bacterial growth. By controlling the water supply by constantly detecting the distortion of the stirring blade caused by the relationship of the above, proper culture is performed for each medium tray, and by using the separation frame, each medium tray can be used as an environment for other medium trays. It is possible to carry out culture suitable for each environment without bothering.

As disclosed in Japanese Patent Application Laid-Open No. 61-128882, the one utilizing the shaft torque of the stirring shaft is used only for monolayer culture, and the strain value of the stirring blade can be detected as in the present invention. For example, it is possible to supply water that matches the environment of each medium tray,
It enables multistage culture.

Next, an apparatus for carrying out the above method will be described.

The culture medium tray (1) of FIG. 1 is stretched with a holding net (2) for storing a solid medium, and a sterile air intake (4) of a peripheral wall (3) corresponding to the lower part of the holding net (2) is opened. There is.

A rotating shaft (5) is erected in the center, and male and female joints (7) and female joints (6) for connecting to a separation frame to be described later are formed on the upper and lower sides. There is a slip ring (8) with a structure that

Further, guide pieces (9) are provided at symmetrical positions on the peripheral wall (3).
(10) is provided in a protruding manner, and each has a guide hole (9a) (10a) fitted to a guide shaft described later.

Furthermore, the rotating shaft (5) has a stirring blade (11) for stirring the solid medium.

The separation frame (12) of FIG. 2 has a separation plate (14) inside the peripheral wall (13), and a water supply nozzle (15) under the separation plate.

From the water supply nozzle (15), in addition to water, a buffer solution for supplying nutrients and controlling the pH of the medium is supplied.

The water supply nozzle (15) includes a siphon nozzle type that sucks and sprays water by air pressure, a spray method that uses a high-pressure pump, and the like, and controls the opening and closing of the supply valve to supply water.

In the separation frame (12), (22) shows an air pipe and (23) shows a water pipe, which are connected to a water supply nozzle (15) which is a two-fluid nozzle.

The peripheral wall (13) has an exhaust port (16), the separation plate (14) has a rotating shaft (17) at the center, and the upper and lower sides of the rotating shaft (17) are the female joints of the medium tray (1) described above. There is a male joint (18) and a female joint (19) connected to (6) and the male joint (7).

The connection end into which the connection end of the slip ring described later is inserted is below the separation plate (14).

And, like the culture medium tray (1), the guide valve (20)
(21) is symmetrically provided on the peripheral wall and has guide holes (20a) and (21a) which fit into guide shafts, which will be described later.

The culture medium tray (1) and the separation frame (12) as described above are alternately stacked in the sterile culture chamber (24), and the joint (7) is fitted to the joint (19) via a slip ring.
The male joint (6) is fitted into the female joint (18) so as to be interlocked.

The medium tray and the separation frame are stacked so that a passage (25) is formed between the uppermost separation frame (12) in the sterile culture chamber (24) and the ceiling of the sterile culture chamber (24). The passage (25) communicates with the outlet (26) of the aseptic culture chamber (24).

Further, below the aseptic culture chamber (24), there is a motor described later, which is connected to the rotary shaft (17) of the lowermost separation frame.

Furthermore, there is a duct (28) on the side of the sterile culture chamber (24), and there are outlets (29) ... (29n) connected to the air intake (4) of the stacked culture medium trays (1). .

There is a partition plate (31) for the duct inner wall (30) to form the outlet (29) ... (29n).

The duct inner wall (30) is formed in a funnel shape together with the partition plate (31) so as to correspond to each stage of the culture medium tray (1), and forms a blowout port (29).

 (32) is a sterile air supply port of the duct (28).

As shown in FIG. 4B, a damper (81) may be provided at the outlet (29) so that sterile air can be adjustably supplied to each stage of the duct (28).

In this case, the sterile air supply port (32) is provided beside the duct (28) as shown in FIG.
2) should be provided to control the total supply of sterile air.

Air pipe (22) of water supply nozzle (15) and water pipe (23)
The structure for joining the flexible pipe (33) to the flexible pipe (33) is as shown in FIG. 6, and a taper surface (41) is formed at the inlet of the air pipe (22) or the water pipe (23). A joint pipe (36) having a tapered surface (42) for engaging with is connected to a conduit (37) communicating with the flexible pipe (33), and the conduit (37) in the bearing (34) of the sterile culture chamber (24). 3
7) slides, and there is a pressing spring (35) between this and the joint pipe (36).

Therefore, if the knob (40) is pulled against the force of the pressing spring (35), the drive rod (39) and the connecting piece (38) are pulled.
The joint pipe (36) is retracted by and the joint with the air pipe (22) or the water pipe (23) is released, and when the knob (40) is released, the joint pipe (36) becomes the air pipe (22) or the water pipe. (23) and their mating surfaces are joined together via an O-ring (43).

 The flexible pipe (33) has a control valve (46).

Flexible tubing (33) in each stage is a common conduit (44)
Common control valve (47) (47a) connected to each (45)
Each is controlled by.

Reference numeral (48) shown in FIG. 7 is a strain gauge attached to the stirring blade (11), and a slip ring (8) capable of sending its signal to an external control device so as to control the supply valve (46). ) Is attached to the upper end of the rotating shaft (5) of the culture medium tray (1) as described above, and the slip ring (8) includes an inner cylinder (50a) that rotates together with the rotating shaft (5).
It consists of a fixed outer cylinder (50b) that comes into contact with this, and the fixed outer cylinder (50b) has a connection terminal (49), which is the separation plate (1).
It is designed to be connected to the connection terminal (27) of 4).

As the strain gauge (48), in addition to FAE foil strain gauge, foil gauge with vinyl cord, paper gauge, semiconductor gauge, plastic range gauge, anti-magnetic gauge, temperature measuring gauge and the like are used.

The slip ring (8) may be a non-contact type by an electromagnetic induction method, as well as a contact type signal transmitting device required for measuring a rotating body as shown in the figure.

The configuration of the medium tray (1) and the separation frame (12) has been described above. An apparatus for stacking these will be described with reference to FIGS.

In the conveyor (58), the medium tray (1) and the separation frame (1
2) and 2) are transferred alternately, but as shown in Figs. 10 and 11, the medium tray (1) is transferred by the conveyor (58) covered with the cover (51), and the medium supply port is in the middle of the transfer. (53)
The culture medium is supplied through the rotary valve (54) of the tank (52) with the fixed medium, and the surface of the fixed culture medium is flattened by the scraping plate (55).

In the shown, only the support shaft of the scraping plate (55) is shown.

The cover (51) communicates with the outside air from the opening duct (56) through the sterilization filter (57).

Further, the culture medium tray (1) and the separation frame (12) are sterilized so that the inside of the cover (51) is aseptic and supplied from a clean room at the entrance of the cover (51).

Medium tray (1) transferred by conveyor (58)
Is taken over by the next chain conveyor (59), but there is a rail (60) parallel to the upper side of the chain conveyor (59) and inside the chain conveyor (59). Push-in pieces for chain fixing (61)
Therefore, the pushing piece (61) moves together with the movement of the chain (59).

So, from the conveyor (58) to the chain conveyor (59)
In the medium tray (1) that is taken over by the guide pieces (9) and (10), the guide pieces (9) and (10) ride on the rail (60). Is pushed to the rear of the rail, and the guide pieces (9) (10) move to the rail (60).
Move while sliding to the left on the diagram.

There are a pair of left and right chain conveyors (59) across the culture chamber (24), and a pair of left and right chain conveyors (59) are connected by a chain drive motor (69) via a bevel gear mechanism (70) and an interlocking shaft (71). Since the rails (60) have a left and right pair corresponding to the chain conveyor (59), the guide pieces (9) and (10) are supported by the rails (60) on both sides, and the medium tray (1). ) Will be transferred in a suspended state.

Incidentally, (62) and (63) are chain wheels of the chain conveyor (59). The rail (60) has a pair of left and right guide shafts (64). Therefore, when the culture medium tray (1) comes to the position of the guide shaft (64), the guide holes of the guide pieces (9) (10). (9a) and (10a) fit to the left and right guide shafts (64).

When mated, the chain conveyor (59) rotates in the reverse direction,
The pushing piece (61) retracts and stands by at a position where it does not interfere with the progress of the next separating frame (12).

When the guide pieces (9) (10) are fitted to the guide shaft (64), the pair of left and right lifting cylinders (65) work, and the push-up piece (68) of the piston rod (67) causes the guide pieces (9) ( 10) Push up.

The medium tray (1) pushed up is pushed up to the position of a pair of left and right height position cylinders (72) shown in FIG. 5, and is supported by the piston rod (73) of the height position cylinder (72). Guide piece (9) with piece (74)
(10) is supported.

Therefore, while the medium tray (1) is being supported,
The lifting piece (68) of the lifting cylinder (65) descends and waits for the next separation frame (12) to come.

The guide pieces (9), (10), (20) and (21) of the medium tray (1) and the separation frame (12) are the lifting cylinder (65).
Two may be provided for each of the cylinders and the high position cylinder (72).

In addition, the medium tray (1) and the separation frame (12) are either mounted on the lifting cylinder (65) or the high position cylinder (72).
It may support the lower part of the peripheral wall (3) (13).

When the separation frame (12) is transferred by the chain conveyor (59) and pushed up by the lifting cylinder (65), the guide pieces (20) (21) are guided by the guide piece (9) of the culture medium tray (1).
Since it is performed in the same manner as (10), when the separation frame (12) rises along the guide shaft (64), the supporting piece (74) is retracted by the height position cylinder (72), and the medium tray ( Since 1) is not supported and is free, it is pushed up by the next separation frame (12).

The separation frame (12) pushing up the culture medium tray (1) is supported by the advancement of the support piece (74) by the operation of the height position cylinder (72).

When the separation frame (12) is supported by the support piece (74), the push-up piece (68) of the elevating cylinder (65) descends and waits for the next medium tray to come.

When the above operation is repeated and the culture medium tray (1) and the separation frame (12) are alternately stacked, the pressing piece (74) does not work on the lowermost separation frame (12) and the guide piece (20). ) (2
While holding 1) up by the pushing piece (68), lower it by the lifting cylinder (65).

Then, the stacked medium trays (1) and the entire separation frame (12) are lowered to the fixed frame (76) shown in FIG. 5 provided on the floor of the sterile culture chamber.

The female joint (19) of the descending lowermost separation frame (12) is fitted to the lower motor shaft (77), and the rotary shafts (5) (7) of the culture medium tray (1) and separation frame (12) are , Interlockingly connected with each other.

 Therefore, the stirring blade (11) can be driven.

Further, the connection terminal (49) of the slip ring (8) provided on the rotary shaft (5) of the culture medium tray (1) has a separation frame (12).
Is connected to the connection terminal (27) provided on the separation plate (14).

The lead wire (80) of the connection terminal (27) has the same structure as the one in which the air pipe (22) or the water pipe (23) shown in FIG. 6 (a) is joined to the external flexible pipe (33). Figure 6 (b)
As described above, the signal from the strain gauge (48) in the stirring blade (11) is sent to the external control system by connecting with the external cable, and the supply valve (46) is controlled.

The reference numeral with a in FIG. 6B is the reference numeral corresponding to FIG.

Reference numerals (83) and (84) denote outlets that fit together.

 The above engagement is performed for each stage.

Furthermore, the drive rod (39) at the connecting portion of the air pipe (22) or the water pipe (23) is retracted at the time of stacking or unloading, but the medium tray (1) and the separation frame (12) as a whole are When it descends to the fixed frame (76), it moves forward, and the drive rod is manually or automatically operated.

Since the guide pieces (9), (10), (20) and (21) are fed in a fixed direction by the guide rod (64), the strain gauge (48) of the strain gauge (48) is lowered when the guide pieces (9), (10), (20) and (21) descend to the fixed frame (76). Leader line (8
0) and the air pipe (22) or the water pipe (23) are correctly joined.

Next, when carrying out the medium tray (1) and the separation frame (12) of each stage from the aseptic culture chamber (24), the above-mentioned operation is performed in reverse.

That is, the lifting cylinder (65) first operates, and the push-up piece (68) pushes the guide pieces (20) (21) of the separation frame (12).
To lift the culture medium tray (1) and the entire separation frame (12), and then the height position cylinder (72) operates to press the guide pieces (9) and (10) of the culture medium tray (1) to hold them. 74)
Support with.

Next, the push-up piece (68) descends, the lowermost separation frame (12) descends, the guide pieces (20) (21) are placed on the rails (60), and the chain conveyor (59) is placed. Reversely push the guide pieces (20) (21) from the front and push them in (61)
To retreat.

The separated separation frame (12) is transferred to the conveyor (58) and carried out to the outside.

In this case, the pushing piece (61) is the guide piece (10).
(9), (20), (21) end (10b) (9b), (20b)
It has a width that pushes only (21b) and protrudes forward and backward from the support shaft (61a) of the chain conveyor (59), so the guide shaft (64) does not get in the way and the chain conveyor (59) is reversed. You can turn around.

Below, the lowering of the push-up piece (68) and the support piece (74) (74)
By moving back and forth, the next medium tray (1) and separation frame (1
2) is unloaded.

As described above, at the stage of transporting the medium tray or the like to the sterile culture room (24), the solid medium is supplied from the tank (52) to the medium tray (1) to separate the medium tray (1) from each other. The frame (12) and the culture medium tray (1) are alternately transported to the sterile culture chamber (24) to stack them.

Blow-off port (29) of the sterile air supply duct (28) to the sterile culture chamber (24) when stacked in the number of design stages of the sterile culture chamber
Is aligned with the air intake (4) of the peripheral wall (3) which is the lower part of the holding mesh (2) of the medium tray (1), and sterile air is supplied from here to the lower part of the medium tray (1), The gas is exhausted toward the inner peripheral wall of the aseptic culture chamber (24) or the exhaust port (16) of the separation frame (12) loaded on the upper part through the space between the solid culture media and rises. This ventilation is performed for each stage.

As described above, the circulating sterile air rises and merges from the passage (25a) of FIG. 4 which is formed between the inner wall (30) and the shelf, and merges from the passage (25) to the discharge port (26) of the sterile culture chamber ceiling. ) Is exhausted from.

For the water supply, since the strain value of the strain gauge (48) of the stirring blade (11) attached to the rotating shaft (5) of the culture medium tray (1) is displaced from the relation with the moisture and friction of the solid culture medium, Is detected and water is sprayed from the water supply nozzle (15) attached to the lower part of the separation plate (14) of the separation frame (12) by controlling the opening / closing of the supply valve (46).

Since the rotary shafts (5) (17) of each stage are interlockingly connected with each other by a joint, the stirring blade (1
Since 1) rotates and agitates the solid medium, it is well ventilated, there is no heat generation due to the growth of microorganisms and the mycelium does not tighten, and the wind pressure does not rise. The temperature and humidity and the air flow rate are adequately adjusted.

Moreover, since there is a separation frame between the culture medium trays, the grounding trays in each stage are completely fractionated and are not controlled by the environment of each other.

Further, the water supply nozzle (15) is attached to the lower surface of the separation plate (14), and at the stage of stacking, the water from the water supply nozzle (15) is sprayed from above toward the culture medium of the culture medium tray (1). Furthermore, since the water content is sprayed from the water supply nozzle (15) while being controlled by the control valve (46) by the strain value detected by the strain gauge (48), an appropriate water supply can be performed. Become.

As described above, solid culture is carried out in the medium tray (1), and after the culture is completed, the medium tray (1) and the separation frame (12) are alternately discharged from the sterile culture chamber (24), and the medium tray ( The target substance will be recovered from 1).

As described above, since the culture medium trays (1) are alternately stacked through the separation frame (12), a large amount of culture can be obtained, and each culture medium tray is equipped with a stirring blade and its strain gauge (48). Since the water supply is controlled for each medium tray, culturing suitable for the culturing conditions is performed,
Since the medium trays are isolated from each other by a separation frame and are not controlled by other medium trays, proper culture is performed.

(Effect of the Invention) The present invention has the following operational effects.

(1) The aeration-type solid culture method is performed in multiple stages to enable large-scale culturing, and the production cost of the target substance can be reduced.

(2) In multi-stage culture, it is possible to aerate each stage and adjust water content for each stage, thereby eliminating imbalance in culture.

(3) The solid medium in multi-stage culture can be agitated, and the environment of culture conditions can be controlled.

(4) Since culture control for each stage can be performed from abnormal changes in strain value and the like, it is difficult to minimize contamination damage due to growth of Bacillus bacteria by medium drying such as water stoppage.

(5) In terms of the device, the grounding tray and the separation frame can be taken out from the sterile culture chamber, so that the device can be easily washed.

In particular, the holding net of the culture medium tray can be easily washed because the bacteria and the culture medium stick to it.

(6) A water supply nozzle attached to the separation frame can supply a nutrient solution and a buffer solution for controlling the pH of the medium, which enables efficient management.

(7) Due to the multi-stage culture, the device can be downsized with respect to the production amount of the target substance.

[Brief description of the drawings]

FIG. 1 (a) (b) is a perspective view of the medium tray and a front view of the half cut, FIG. 2 (a) (b) is a perspective view of the separation frame and a front view of the half cut, and FIG. FIG. 4 is a plan view of the apparatus, FIG. 4 (a), (b), and (c) are the same as the above AA sectional view and the partial sectional view and the partial plan view of the modified embodiment. FIG. 5 is the same as the above BB sectional view. Figures (a) and (b) are desorption structure diagrams of the water pipe and air pipe and the desorption structure diagram of the strain gauge lead wire. Fig. 7 is an enlarged view of the strain gauge portion. Fig. 8 is a cross-sectional view of the slip ring portion. FIG. 10 is an explanatory view of the stacking device for the culture medium tray and the separation frame as seen from the plane and front, FIG. 11 is a perspective view showing the relationship between the medium tray and the loading / unloading device for the separation frame, and the sterile culture chamber, and FIG. It is a partially expanded perspective view of a stacking device. (1) …… Medium tray (2) …… Retaining net (3) …… Peripheral wall (4) …… Aseptic air supply inlet (5) …… Rotating shaft (8) …… Slip ring (9) (10) ...... Guide piece (12) …… Separation frame (13) …… Peripheral wall (14) …… Separation plate (15) …… Moisture supply nozzle (16) …… Exhaust port (17) …… Rotary shaft (20) ( 21) …… Guide piece (22) …… Air tube (23) …… Water tube (24) …… Sterile culture chamber (28) …… Duct (29) …… Blowout port (32) …… Sterile air supply port ( 46) Supply valve (48) Strain gauge (58) Conveyor (59) Chain conveyor (60) Rail (61) Push piece (64) Guide shaft (65) … Lifting cylinder (68) …… Pushing up piece (72) …… Height position cylinder (74) …… Supporting piece (76) …… Fixed frame

Claims (5)

(57) [Claims] 1. A medium tray for storing a solid medium is stacked in multiple stages in a sterile culture chamber to which sterile air is supplied through a separation frame having a water supply nozzle for supplying water to the solid medium. While stirring the medium with a stirring blade,
Aseptic air is supplied to each stage from the bottom of each medium tray and discharged from the top, and moisture is supplied from the moisture supply nozzle of each separate separation frame toward the solid medium in the medium tray of each stage. A multi-stage solid culture method comprising culturing while controlling an aseptic air and water while setting an environment according to the culture condition of the stage. 2. The water supply is controlled by detecting the strain value of the stirring blade that is caused by the relationship between the stirring blade that stirs the solid medium and the friction coefficient between the medium and the medium due to the growth of bacteria and bacteria. The described multistage solid culture method. 3. A solid medium holding network for storing the solid medium,
A solid medium in the medium tray, which has a rotatable stirring blade for stirring the solid medium, and a medium tray having a sterile air inlet on the lower peripheral wall of the holding mesh, and a sterile air exhaust port on the frame peripheral wall. A separation frame equipped with a water supply nozzle that supplies water to the medium is alternately stacked in multiple stages in a sterile culture chamber, and the rotation shafts of the stirring blades of each medium tray can be interlockingly connected to each other via the rotation shaft of the separation frame. Multi-stage solid culture device. 4. A strain gauge attached to a stirring blade of a culture medium tray and a signal sent to a moisture control device via a slip ring, and the output signal is output to a supply valve of a moisture supply nozzle to supply a feed valve. The multistage solid culture device according to claim 3, wherein the ON / OFF control is performed. 5. A guide shaft fitted to each guide piece of a culture medium tray and a separation frame and vertically guiding them to a predetermined direction and a fixed position, and a culture chamber inlet to the tray and the separation frame to the guide shaft. A guide rail for moving the tray and the separating device along with the guide device for guiding the tray and the separating frame, and a cylinder device for vertically moving the tray and the separating frame along a guide axis; and the tray. The multi-stage solid culture device according to claim (3), wherein the culture medium tray is provided with a cylinder device that temporarily holds the separation frame at a constant height, and the separation frame is stacked.