JPH0563300U - Incubator - Google Patents

Abstract

(57) [Abstract] [Purpose] To cultivate a relatively large amount of microorganisms and plant cells efficiently. [Structure] A granular gel or plant cells to which microorganisms are adhered is put inside a porous support frame 37 made of a wire mesh or the like.
The culture solution taken out from the culture solution outflow pipe 31 from the storage cylinder 26 is passed through the culture solution inflow pipe 32 and blown out by a nozzle
Send in. Nozzle holes 35, 3 of the blowing nozzle 34
The culture solution blown out from 5 flows from the inside to the outside of the support frame 37 to attach the granular gel and the plant cells to the inner peripheral surface of the support frame 37. Since the flow of the culture solution is averaged by the baffle cylinder 44, the granular gel and plant cells adhere well to the support frame 37.

Description

[Detailed description of the device]

[0001]

[Industrial applications]

 The culture device according to the present invention can be used for culturing microorganisms such as yeast or lactic acid bacteria, or plant cells.

[0002]

[Background of the invention]

 With the development of biotechnology, it has become common to culture various microorganisms, and along with this, various new culture methods are being considered. In order to carry out efficient culture, it is necessary to constantly stir the culture medium so that the culture medium can be brought into good contact with the microorganisms or plant cells to be cultured.

As a culture device used in such a case, for example, Japanese Patent Publication No. 2-40316 discloses a culture device as shown in FIG.

In this conventionally known culture device, a disc-shaped bottom plate 2 having an outer diameter slightly larger than the outer diameter of the glass tube 1 is fixed to the lower end of the cylindrical glass tube 1. However, the lower end opening of the glass tube 1 is closed. Lower end portions of a plurality of studs 3, 3 are respectively screwed and fixed to a portion of the bottom plate 2 near the outer periphery of the bottom plate 2 that protrudes outward from the outer peripheral surface of the glass tube 1. The upper ends of the studs 3 and 3 respectively project slightly above the upper edge of the glass tube 1, and the upper ends of the studs 3 and 3 are disc-shaped having an outer diameter slightly larger than the outer diameter of the glass tube 1. They are inserted into circular holes 5 and 5 formed in the outer peripheral portion of the lid body 4, respectively. Nuts (not shown) can be screwed onto the upper ends of the studs 3 and 3 projecting from the upper surface of the lid body 4 through the circular holes 5 and 5, so that the lid body 4 is attached to the upper end of the glass tube 1. It is attached to the opening so that it can be opened and closed.

Inside the glass tube 1, a horizontal circular disk 23 is provided substantially concentrically with the glass tube 1, and a straightening plate 25 is provided on the inner peripheral edge of the circular disk 23 for stays 7, 7. It is fixed by. The current plate 25 has a cylinder 24 that rises upward from the inner peripheral edge of the circular plate 23.

At the lower end of the rectifying plate 25, a wire mesh formed by braiding stainless steel filaments is formed into a cylindrical shape, and the lower end of the support frame 6 is externally fitted, so that the support frame 6 is attached to the glass tube. The glass tube 1 is fixed to the inside of the glass 1 substantially concentrically.

On the other hand, a vertical shaft 8 is planted in the center of the upper surface of the bottom plate 2, and an inverted cylindrical cap 10 fixed to the center of a rotary block 9 is rotatably attached to the vertical shaft 8. I'm wearing it. An annular magnet 11 is fitted and fixed around the rotating block 9, and the magnet 11 and the annular magnet 16 are opposed to each other with the bottom plate 2 interposed therebetween. The annular magnet 16 is fixed to the output shaft 15 of the motor 14, which is fixed via a bracket 13 below the base plate 12 to which the bottom plate 2 is fixed.

A plurality of flat blades 17, 17 are radially fixed to the outer periphery of the rotary block 9 rotatably supported by the vertical shaft 8 via a cap 10, thereby enabling centrifugal separation. The wing fan 18 is configured.

Further, above the centrifugal multi-blade fan 18, an air bubble nozzle for blowing air into the liquid stored inside the glass tube 1 is provided inside the cylinder 24 constituting the flow straightening plate 25. 19 are provided. The bubble nozzle 19 has a plurality of small holes 21, 21 at the center of a lid 4 that opens and closes the upper end opening of the glass tube 1 and at the lower end of an air pipe 20 fixed in a state of penetrating the lid 4. The nozzle portion 22 is fixed, and the compressed gas fed through the air feeding pipe 20 is ejected to a position directly above the centrifugal multiblade fan 18. Although illustration and detailed description are omitted, the lid 4 is provided with a sensor for detecting the state of the culture solution in the glass tube 1 in addition to the air supply tube 20, and a sensor for feeding nutrients into the culture solution. Pipes etc. are provided.

When culturing a microorganism such as yeast or lactic acid bacterium, or a plant cell by the conventional culturing apparatus configured as described above, first, the culturing solution should be cultivated inside the glass tube 1 together with the culturing solution. A granular gel with microorganisms fixed and plant cells to be cultured are added.

When the charging is completed, compressed air is sent into the nozzle part 22 constituting the bubble nozzle 19 through the air supply pipe 20, and the compressed air is stored in the glass cylinder 1 through the small holes 21 and 21 of the nozzle part 22. The centrifugal multi-blade fan 18 is rotated by energizing the motor 14 while blowing air into the culture medium.

That is, when the magnet 16 fixed to the output shaft 15 of the motor 14 is rotated by energizing the motor 14, the magnet 11 facing the magnet 16 via the bottom plate 2 is generated by the magnet coupling action. A centrifugal multi-blade fan 18 composed of a plurality of blades 17 and 17 which rotates and is fixed to the rotating block 9 together with the magnet 11 is provided below the circular plate 23 constituting the flow straightening plate 25. It rotates about the vertical axis 8.

The rotation of the centrifugal multi-blade fan 18 accompanying the energization of the motor 14 causes the bottom portion of the glass tube 1 in which the culture solution is stored to be located near the center of the glass tube 1 along the lower surface of the circular plate 23. The flow inside the support frame 6 communicating with the inside of the cylinder 24 forming the flow regulating plate 25 becomes lower than the pressure outside the flow.

When a pressure difference occurs inside and outside the support frame 6 as the centrifugal multiblade fan 18 rotates in this way, the culture solution existing outside the support frame 6 flows into the support frame 6. Another flow is induced, and the culture solution in the glass tube 1 flows down inside the support frame 6 and rises in a tubular space between the outer peripheral surface of the support frame 6 and the inner peripheral surface of the glass tube 1. While flowing, it circulates in the glass tube 1. However, a large number of small holes (mesh) formed in the support frame 6 present in the middle of this flow are due to the large number of granular gels and plant cell particles (3 to Smaller than 5 mm). Therefore, each granular gel that tends to flow with the culture solution cannot pass through these small holes, and remains attached to the outer peripheral surface of the support frame 6, and only the culture solution flows while passing through the support frame 6. ..

As described above, in the culture fluid flowing through the support frame 6 having the outer peripheral surface to which the granular gel is attached, the inner side of the cylinder 24 forming the straightening plate 25 is directly above the centrifugal multiblade fan 18. Since air is blown in from the bubble nozzle 19 provided in the chamber, and a sufficient amount of air is always dissolved in this culture solution, the microorganisms fixed in the granular gel attached to the support frame 6 are fixed. Is constantly supplied with sufficient air, continuous culture is performed, and the number of microorganisms in the culture solution gradually increases.

[0016]

[Problems to be solved by the device]

 However, in the case of the conventional culture device configured and operating as described above, it is difficult to increase the size, and it is not possible to culture a large amount of microorganisms or plant cells at one time.

That is, in the case of the above-mentioned conventional structure, the centrifugal multi-blade fan 18 was used to induce the flow of the culture solution through the support frame 6, but with the increase of the capacity of the culture device. When the diameter of the support frame 6 becomes large, it becomes difficult to induce a uniform flow in the entire support frame 6, and the granular gel or plant cells to which the microorganisms to be cultivated adhere are Only the parts will adhere. Further, the rotation of the centrifugal multi-blade fan 18 causes a vortex flow of the culture solution, and the vortex flow applies a shearing force to the microorganisms or plant cells to be cultured. As a result, the efficiency of the culturing work is reduced.

The culturing apparatus of the present invention was devised in view of the above circumstances.

[0019]

[Means for solving the problem]

 The culture device of the present invention has a lower end opening closed by a bottom plate and a storage tube provided with an openable lid at the upper end opening, and a storage tube vertically arranged around the center of the storage tube and provided on the peripheral surface. A plurality of nozzle holes are provided so that the culture solution fed into the inside can be spouted from the nozzle holes to the surroundings, and inside the storage cylinder, around the blowing nozzle, almost concentric with the blowing nozzle. The cylindrical support frame provided and the culture existing inside the storage cylinder, which is arranged around the support frame and substantially concentric with the blowout nozzle, through a plurality of holes provided in the peripheral surface. It is composed of an outer cylinder that allows the liquid to flow out to the surroundings, and a pump for sending the culture liquid that has flowed out to the outer peripheral side of the outer cylinder to the inside of the blowing nozzle.

[0020]

[Action]

 When culturing microorganisms or plant cells with the culture device of the present invention configured as described above, the culture solution is put into the storage cylinder and the space between the inner peripheral surface of the support frame and the outer peripheral surface of the blowing nozzle is set. The pump is operated in a state where the granular gel or plant cells to which the above microorganisms are attached is put into the cylindrical space.

With the operation of the pump, the culture solution existing on the outer side of the outer cylinder is sucked into the pump and is ejected from the nozzle hole of the blowing nozzle.

As a result, the culture solution flows from the inside of the support frame to the outside, and the granular gel and plant cells injected into the cylindrical space adhere to the inner peripheral surface of the support frame. The flow of the culture solution from the inner side of the support frame to the outer side of the outer tube becomes uniform due to the presence of the outer tube, and this uniform flow does not stop as long as the operation of the pump is continued. Therefore, the microorganisms and plant cells attached to the granular gel are continuously exposed to the culture solution, and the microorganisms and plant cells are efficiently cultured.

[0023]

【Example】

 1 to 5 show an embodiment of the present invention. For example, an outer cylinder 27 is attached in a liquid-tight manner to the lower outer peripheral surface of a storage cylinder 26 having a capacity of about 60 liters, and the inner peripheral surface of the outer cylinder 27 and the outer periphery of the storage cylinder 26 are A jacket 28 is formed between it and the surface. Numerals 29 and 30 are pipe connection ports through which steam for sterilization or a fluid for temperature control flows through the jacket 28. Reference numeral 31 is a culture solution outflow pipe provided at two positions on the lower outer peripheral surface of the storage tube 26, and 32 is a culture solution inflow pipe provided so as to penetrate the central portion of the bottom plate 33 in a liquid-tight manner.

In a continuous state above the culture solution inflow pipe 32, a circular tube-shaped blow-out nozzle 34 having a closed upper end is provided in the central portion of the storage cylinder 26. A plurality of nozzle holes 35, 35 are formed vertically in the blowing nozzle 34 so that the culture solution sent from the culture solution inflow pipe 32 is ejected toward the surroundings.

Around the blow-out nozzle 34, a support frame 37, which is formed of a wire mesh into a bottomed cylindrical shape having a capacity of about 30 liters, is provided. That is, the support frame 37 includes a cylindrical portion 38 and a circular ring-shaped bottom plate portion 39 that closes the lower end opening of the cylindrical portion 38 except for the circular hole provided in the central portion. By fitting the circular hole of the bottom plate portion 39 into the blowing nozzle 34, the supporting frame 37 is supported around the blowing nozzle 34. In this state, the blowing nozzle 34 and the cylindrical portion 38 are concentric with each other.

In the illustrated embodiment, the lower end of the blowing nozzle 34 and the upper end of the culture solution inflow pipe 32 are liquid-tightly fitted by interposing an O-ring 42 between them. The culture solution inflow pipe 32 and the blowing nozzle 34 are connected. Further, at the two positions of the upper edge of the cylindrical portion 38 constituting the support frame 37, tiltable handles 43, 43 are provided, and the support frame 37 is pulled from the storage tube 26 by the handles 43, 43. I am making it available.

On the outside of the support frame 37, a cylindrical baffle cylinder 44 made of a perforated plate, which is an outer cylinder, is provided. That is, the support ring 45 is welded and fixed to the upper end edge of the baffle cylinder 44, and the support ring 45 is fixed to the inner peripheral surface of the upper end edge of the storage cylinder 26. The baffle cylinder 44 has a function of equalizing the flow of the culture solution when circulating the culture solution.

An annular sensor holder 47 is provided on one side of the storage tube 26, and the temperature, PH value, and dissolved oxygen concentration of the culture solution in the storage tube 26 are measured in the sensor holder 47. A sensor for this can be inserted freely. A viewing window 48 is provided on the other side surface of the storage tube 26, and the inside of the storage tube 26 can be viewed through the viewing window 48. A lighting window 49 (FIGS. 3 to 4) is provided on the upper side surface of the storage cylinder 26 so that the inside of the storage cylinder 26 can be illuminated.

On the other hand, the upper end opening of the storage cylinder 26 can be opened and closed by the lid 51. The handle 52 is fixed to one end (the left end in FIG. 2) of the upper surface of the cover 51, and the cover 51 is attached to the other end (the right end in FIG. 2) of the upper surface by operating the lever 78. A lifter 77 that lifts a little is provided. Further, U-shaped cuts 53, 53 are formed at equal intervals at a plurality of locations (six locations in the illustrated example) on the periphery of the lid 51. Base portions of bolts 54, 54 are pivotally attached to portions of the outer peripheral surface of the upper end portion of the storage cylinder 26 which can be aligned with the notches 53, 53, respectively, and nuts 55 are provided at the tip portions of the bolts 54, 54. , 55 are screwed together.

When the upper end opening of the storage cylinder 26 is closed, with the bolts 54, 54 hanging downward, the opening 51 is covered with the lid body 51, and the notches 53, 53 and the bolts. After aligning the bolts 54, 54 with each other, the bolts 54, 54 are rotated upward so that the intermediate portions of the bolts 54, 54 are inserted into the notches 53, 53. Next, the nuts 55, 55 screwed onto the tips of the bolts 54, 54 are tightened, and the lower surface of the nuts 55, 55 holds the lid 51 against the upper edge of the storage tube 26. When opening the upper end opening described above, the reverse operation is performed, and then the lever 78 which constitutes the lifter 77 is operated to slightly lift the lid body 51 by the lifter 77 and further hold the handle 52 to hold the vertical shaft. The lid 51 is rotated laterally around the center.

As described above, the lid 51 attached to the upper end opening of the storage cylinder 26 is provided with the viewing windows 56, 56, the flame inoculation port 57, the exhaust port 59, and the pressure gauge 60. ing.

Further, as schematically shown in FIG. 5, the culture solution outflow pipe 31 and the culture solution inflow pipe 32 are connected by a circulation pipe 66 provided with a solution sending pump 65 in the middle thereof. A supply pipe 79 and a drainage pipe 74 are connected in the middle of the circulation pipe 66, and air or water vapor is sent into the storage cylinder 26 through the supply pipe 79, the circulation pipe 66, and the culture solution inflow pipe 32. The culture solution can be freely discharged through the culture solution outflow pipe 31, the circulation pipe 66, and the drainage pipe 74. Further, one end of a culture solution supply pipe 50 and a chemical solution pipe 58 are connected to the circulation pipe 66, and the culture solution or various chemical solutions can be freely fed into the storage cylinder 26 through these pipes 50, 58. There is.

On the other hand, a steam supply pipe 64 having a valve 62 in the middle is connected to the upper side surface of the jacket 28, and a heater 67 and a valve 68 are provided in the middle of the lower side surface of the jacket 28. One end of the water pipe 69 is connected. In the middle of the steam supply pipe 64 and the water pipe 69, a drain discharge pipe 72 provided with a valve 70 and a steam trap 71 is connected to the jacket 28 side of the valves 62 and 68, respectively. The steam or moisture in the jacket 28 can be discharged freely.

Further, around the lower end portion of the blowing nozzle 34, an air supply pipe 40 is provided for uniformly dispersing the microorganisms to be cultured in the support frame 37 together with the culture solution. The air supply pipe 40 formed in an annular shape is connected to an air source outside the storage cylinder 26 by a pipe (not shown), and air supply holes 41, 4 provided on the upper surface of the air supply pipe 40 in advance before starting the culture work. By sending out air from 1, the above microorganisms are dispersed together with the culture solution.

Further, an exhaust port 76 provided with a valve 75 in the middle is connected to the exhaust port 59 provided in the lid body 51 so that the gas existing in the storage cylinder 26 can be discharged freely.

When culturing microorganisms or plant cells using the culturing apparatus of the present invention configured as described above, work is performed as follows.

First, with the lid 51 closing the upper end opening of the storage cylinder 26, the valve 61 in the middle of the steam supply pipe 63 is opened, and the supply pipe 79, the circulation pipe 66, and the culture solution inflow pipe 32 are inserted. Then, high temperature steam is passed through the storage cylinder 26 to sterilize the inside of the storage cylinder 26. The condensed water generated in the storage cylinder 26 is discharged through the culture solution outflow pipe 31, the circulation pipe 66, and the drainage pipe 74.

After the sterilization for a predetermined period of time, the valve 61 is closed, and then the culture solution is injected into the storage tube 26 through the culture solution supply pipe 50 connected to the circulation pipe 66. Next, the valve 62 in the middle of the steam supply pipe 64 is opened, high-temperature steam is sent to the jacket 28, and the culture solution in the storage tube 26 is heated to an appropriate temperature. Next, the valve 68 in the middle of the water pipe 69 is opened, and the water heated by the heater 67 is supplied to the jacket 28 to keep the culture solution in the storage tube 26 at an appropriate temperature. Condensed water and the like generated in the jacket 28 is separately discharged from the drain discharge pipe 72 by opening the valve 70.

When the culture solution in the storage tube 26 reaches an appropriate temperature, the microorganisms and plant cells to be cultured through the flame inoculation port 57 are put in the storage tube 26 and inside the support frame 37, and then, Air is sent out from the air supply holes 41, 41 of the air supply pipe 40 to uniformly disperse the microorganisms and plant cells in the support frame 37. Then, the liquid feed pump 65 is operated to circulate the culture solution through the circulation pipe 66. That is, with the operation of the liquid feed pump 65, the culture liquid in the storage tube 26 is discharged from the culture liquid outflow pipe 31, and is fed into the blowing nozzle 34 through the culture liquid inflow pipe 32.

The culture solution thus fed into the inside of the blowing nozzle 34 then flows out into the support frame 37 from the nozzle holes 35 of the blowing nozzle 34. As a result, the flow of the culture solution flowing from the inside to the outside of the support frame 37 is induced, and the microorganism-attached granular gel or plant cells introduced into the support frame 37 are the above-mentioned support frame 37. It is exposed to the flow of the culture solution while being attached to the inner peripheral surface of the. As a result, the above microorganisms and plant cells grow while absorbing the nutrients in the culture solution.

During the culturing operation, the temperature of the culture solution in the storage tube 26 is adjusted by adjusting the temperature of the water flowing into the jacket 28 through the heater 67, and the gas generated during the culturing is controlled by the pressure gauge 60. While watching, the valve 75 provided in the middle of the exhaust pipe 76 is opened and discharged at appropriate times.

The state where microorganisms and the like propagate in the storage tube 26 can be seen by looking through the observation windows 48 and 56 into the storage tube 26, and changes in temperature, PH value, etc. can be detected from the sensor holder 47 to the storage tube 26. It can be known by the sensor inserted inside, and when these values deviate from the desired values, it is possible to inject the chemical solution from the chemical solution pipe 58 and adjust them.

When the culture is completed, the lid 51 is removed, the handles 43, 43 are held, and the support frame 37 is lifted. As a result, the support frame 37 is pulled up from the storage cylinder 26, and the microorganisms and plant cells that have propagated and filled in the support frame 37 can be taken out.

[0044]

[Effect of the device]

 Since the culture device of the present invention is configured and operates as described above, it does not destroy a relatively large amount of granular gel or plant cells mixed in the culture medium, and the culture medium and granular gel or plant cells are not destroyed. It is possible to make good contact with the microorganisms and efficiently culture microorganisms.

[Brief description of drawings]

FIG. 1 is a front view showing an embodiment of the present invention.

FIG. 2 is a plan view of the same.

FIG. 3 is a vertical sectional view of a storage cylinder.

FIG. 4 is a transverse sectional view of the same.

FIG. 5 is a schematic circuit diagram.

FIG. 6 is a vertical sectional side view showing a conventional structure.

[Explanation of sign]

 1 Glass Cylinder 2 Bottom Plate 3 Stud 4 Lid 5 Circular Hole 6 Support Frame 7 Stay 8 Vertical Axis 9 Rotating Block 10 Cap 11 Magnet 12 Board 13 Bracket 14 Motor 15 Output Shaft 16 Magnet 17 Wing Plate 18 Centrifugal Multiblade Fan 19 Bubble Nozzle 20 Air Supply Pipe 21 Small Hole 22 Nozzle Part 23 Circular Plate 24 Cylinder 25 Rectifier Plate 26 Storage Cylinder 27 Outer Cylinder 28 Jacket 29 Tube Connection Port 30 Tube Connection Port 31 Culture Solution Outflow Tube 32 Culture Solution Inflow Tube 33 Bottom Plate 34 Blowout Nozzle 35 Nozzle hole 37 Support frame 38 Cylindrical part 39 Bottom plate part 40 Air supply pipe 41 Air supply hole 42 O-ring 43 Handle 44 Baffle cylinder 45 Support ring 47 Sensor holder 48 Viewing window 49 Daylighting window 50 Culture solution supply pipe 51 Lid body 52 Handle 53 Notch 54 Bolt 55 Nut 56 Viewing window 57 Flame injection port 58 Liquid pipe 59 Exhaust port 60 Pressure gauge 61 Valve 62 Valve 63 Steam supply pipe 64 Steam supply pipe 65 Liquid feed pump 66 Circulation pipe 67 Heater 68 valve 69 Water pipe 70 valve 71 Trap 72 Drain discharge pipe 73 valve 74 Discharge pipe 75 valve 76 Exhaust pipe 77 Lifter 78 Lever 79 Supply pipe

Claims (1)

[Scope of utility model registration request] 1. A storage cylinder having a lower end opening closed by a bottom plate, and an upper end opening provided with a lid that can be opened and closed, and a storage cylinder arranged vertically in the center of the storage cylinder and having a plurality of peripheral surfaces. A nozzle provided with a nozzle hole so that the culture solution fed into the nozzle can be ejected to the surroundings from the nozzle hole, and a cylinder provided inside the storage cylinder around the nozzle and substantially concentric with the nozzle. -Shaped support frame, around the support frame inside the storage cylinder, arranged substantially concentric with the blowing nozzle, through a plurality of holes provided in the peripheral surface, the culture solution present inside, A culture device comprising an outer cylinder that is allowed to flow out to the surroundings and a pump for sending the culture solution that has flowed out to the outer peripheral side of the outer cylinder to the inside of the blowing nozzle.