JP2022011205A - Stirrer-free biological reactor using fine bubbles and ultra fine bubbles containing gases useful for culturing microorganisms or cells and biological reaction method using this biological reactor - Google Patents

Abstract

To provide a biological reactor and a biological reaction method, each of which uses fine gas bubbles containing a useful gas and is able to sufficiently stir a biological culture solution contained in a culture tank without using a stirrer and to maintain the activity of microorganisms or the like.SOLUTION: When a biological culture solution is extracted from a culture tank to be allowed to contain fine gas bubbles and refluxed to the culture tank, a multi-nozzle provided with a plurality of discharge ports discharges the biological culture solution containing fine gas bubbles containing the useful gas in multiple directions in the culture tank.SELECTED DRAWING: Figure 1

Description

The present invention uses a biological reaction device for culturing a microorganism or a cell (hereinafter, also referred to as "microorganism or the like") to cause the microorganism or the like to produce a reaction product, or a biological reaction device for multiplying the microorganism or the like, and the biological reaction device. The biological reaction method used is characterized in that the biological culture solution containing the culture solution and microorganisms (hereinafter, also referred to as “biological culture solution”) contained in the culture tank is stirred without using a stirrer. Is to be.

Unlike chemical reactions, biological reactions are slow, but they do not use a lot of energy or many chemical substances, so they are mild and meaningful reactions for the environment.

However, the biological reaction has a problem that the reaction is generally slow. That is, in many cases, a reaction within one hour is sufficient for a chemical reaction, whereas in the case of a biological reaction, it may take several hours to several days if it is long, or several weeks or more if it is particularly long. be. Therefore, it is required to carry out biological reactions efficiently and economically.

The present inventors propose in Patent Documents 1 to 4 and the like to efficiently and economically carry out biological reactions of microorganisms and the like by using micro-nano bubbles of oxygen-containing gas.

In the claims and the specification of this case, the conventional "micro bubbles" and "nano bubbles" are referred to as "fine bubbles" and "ultra fine bubbles", respectively.

However, in the biological reaction apparatus and biological reaction method of Patent Documents 1 to 4, the biological culture solution contained in the culture tank is agitated by using a stirrer.
a) Under stress and damage, the activity of microorganisms, etc. is reduced or the growth is inhibited.
b) There are places where it is difficult to sterilize the medium before the biological reaction (seal part of the rotating shaft of the stirrer, bearing part of the rotating shaft, stirring blade, baffle, obstruction plate, etc.) to prevent contamination. Is difficult,
c) Problems such as costly installation, operation, maintenance and management of the stirrer occur.

Further, Patent Documents 5 and 6 propose a method for culturing microorganisms and the like without using a stirrer. However, in Patent Document 5, in a special culture of fed-batch culture of cells (increasing the amount of medium as the cells grow), a special structure is adopted in which the cells are provided in the main body of the culture tank. Natural circulation flow is performed by utilizing the difference in specific gravity of the liquid inside and outside the cylinder. Further, in Patent Document 6, cells are cultured by using a special reaction tank called a reaction tank in which the lower parts of a pair of vertical culture cylinders are connected, and gas is alternately blown from the gas blowing means connected to the lower parts of both culture cylinders. Stirring is performed by a special method of blowing and stirring the culture solution. As described above, in the stirring method disclosed in Patent Documents 5 and 6, the structure in the culture tank becomes complicated and it is difficult to prevent the contamination of various germs. Therefore, a biological reaction device / biological reaction using a general culture tank. It cannot be adopted by the method.

The present inventors have found the following matters and have made the present invention.
1) In the conventional biological reaction device / biological reaction method, the stirrer has the purpose of i) finely shearing the bubbles of the gas supplied to the culture tank, and ii) the purpose of uniformly mixing the biological culture solution in the culture tank. However, in the biological reaction apparatus / biological reaction method using fine bubbles / ultrafine bubbles of oxygen-containing gas as in Patent Documents 1 to 4, the purpose of i) above has already been sufficiently achieved. Therefore, if the purpose of ii) above can be achieved, there is little need to use a stirrer.
2) The purpose of ii) is to make the biological culture solution containing the fine bubbles / ultra fine bubbles refluxed into the culture tank 0 ° to 90 ° with respect to the culture tank side direction of the central axis. It can be achieved economically and efficiently by discharging toward the culture tank using a multi-nozzle equipped with a plurality of discharge ports provided so as to discharge in the direction.
3) As the gas contained in the fine bubble / ultra fine bubble, a gas containing oxygen is used in the culture of <aerobic or permeable anaerobic microorganisms, etc.> as in Patent Documents 1 to 4, but the gas is limited to this. However, in the culture of <obligate anaerobic microorganisms, etc.>, a gas containing nitrogen is used, and organic substances (amino acids, organic acids, proteins, etc.) are used from carbon gas (carbon dioxide gas, methane gas, etc.) derived from natural gas. In the cultivation of the generated microorganisms <organic synthetic microorganisms, etc.>, carbon gas derived from natural gas such as carbon dioxide gas and methane gas can be used, and similarly, the cultivation can be performed without using a stirrer.

The biological reaction apparatus of the present invention and the biological reaction method using this biological reaction apparatus, which were made based on the ideas of 1) to 3) above, were prepared by adding air, oxygen, and the like to the biological culture solution without using a stirrer. Fine bubble of a gas containing a gas useful for culturing microorganisms such as nitrogen, carbon dioxide, carbon gas derived from natural gas, and carbon dioxide gas discharged from a thermal power plant (hereinafter, also referred to as "useful gas"). It is characterized in that a biological culture solution containing ultrafine bubbles (hereinafter, also referred to as "fine bubbles") is discharged in multiple directions in a culture tank using a multi-nozzle. As a result, the problems a) to c) can be solved without reducing the activity of microorganisms and the like, and by using the multi-nozzle, stirring can be performed economically and efficiently.

Japanese Patent No. 5985114 Japanese Patent No. 6087476 Japanese Patent No. 6138390 Japanese Patent No. 6499203 Japanese Unexamined Patent Publication No. 6-327460 Japanese Unexamined Patent Publication No. 2012-115232

The biological reaction apparatus of the present invention (hereinafter, also referred to as "biological reaction apparatus of the present invention") and the biological reaction method using this biological reaction apparatus (hereinafter, also referred to as "biological reaction method of the present invention", are collectively referred to as "biological reaction apparatus of the present invention". The subject of the biological reaction apparatus / method of the present invention is that the biological culture solution contained in the culture tank can be sufficiently stirred without using a stirrer, the activity of microorganisms and the like can be maintained, and the stirrer can be used. By not using, the internal structure of the culture tank / biological reaction device can be simplified, the cleanability can be improved and the contamination by various germs can be prevented, and the organisms that can be stirred economically and efficiently. The purpose is to provide a reaction device and a biological reaction method.

In order to solve the above-mentioned problems, in the biological reaction apparatus / method of the present invention, when the biological culture solution is extracted from the culture tank, the fine bubbles are contained and the mixture is returned to the culture tank, the fine bubbles of the gas containing the useful gas are generated. It is characterized in that the biological culture solution is agitated without using a stirrer by discharging the contained biological culture solution in multiple directions in the culture tank using a multi-nozzle. Specifically, it is arranged in a culture tank for accommodating the biological culture solution, an extraction conduit for extracting the biological culture solution from the culture tank, and between the extraction conduit and the culture tank, and the biological culture solution is returned to the culture tank. The multi-nozzle is a biological reaction device provided with a multi-nozzle, the above-mentioned multi-nozzle is a fine bubble generator for containing fine bubbles of a gas containing a useful gas in a biological culture solution, and a plurality of devices provided at outlets on the culture tank side. A biological culture solution containing fine bubbles (hereinafter, also referred to as “fine bubble-containing biological culture solution”) is supplied from a plurality of discharge ports toward the culture tank side of the central axis of the multi-nozzle. It is characterized by discharging in multiple directions of 0 ° to 90 °.

In the present invention, as described above, a stirrer is used by ejecting the biological culture solution containing the fine bubbles of the gas containing the useful gas in multiple directions in the culture tank using a multi-nozzle. Without doing so, the biological culture solution contained in the culture tank can be sufficiently stirred, and the activity of microorganisms and the like can be maintained.

Further, in the present invention, by not using the stirrer as described above, the internal structure of the culture tank / biological reaction apparatus can be simplified, the detergency can be improved, and the contamination with various germs can be prevented.
By not using the stirrer, a) the microorganisms are stressed and damaged and the activity of the microorganisms is reduced, and b) the part where sterility is difficult to be performed before the biological reaction (the seal part of the rotating shaft of the stirrer, rotation). Shaft bearings, stirring blades, baffles, baffles, etc.) are generated and it is difficult to prevent contamination, c) Installation, operation, maintenance, management, etc. of the stirring machine are costly. You can solve the problem.

As described above, by using the multi-nozzle without using the stirrer, the biological culture solution contained in the culture tank can be agitated economically and efficiently.

It is a schematic diagram which shows the outline of 1st Embodiment of the biological reaction apparatus | method of this invention. It is a schematic diagram which shows the outline of the 2nd Embodiment of the biological reaction apparatus | method of this invention. It is sectional drawing which shows the outline of the multi-nozzle used in the biological reaction apparatus | method of this invention. It is sectional drawing which shows the XX'cross section of FIG. It is a schematic appearance view which shows the left end surface of FIG. It is a schematic diagram for demonstrating the angle on the acute angle side formed by the transport line and the discharge line in the biological reaction apparatus / method of this invention. It is a schematic diagram for demonstrating the angle on the acute angle side formed by the transport line and the discharge line in the biological reaction apparatus / method of this invention. It is a schematic diagram which shows the outline of the 3rd Embodiment of the biological reaction apparatus | method of this invention. It is sectional drawing which shows the fine bubble generator used in the biological reaction apparatus | method of this invention. It is an external schematic diagram which shows the fine bubble generator suitablely used for the biological reaction apparatus | method of this invention. It is sectional drawing which shows the operation state of the fine bubble generator. It is a schematic diagram which shows the conventional biological reaction apparatus.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings, but the present invention is not limited thereto.

<General matters of the biological reaction apparatus / method of the present invention>
First, general matters of the biological reaction apparatus / method of the present invention will be described.

The biological reaction apparatus / method of the present invention can be suitably used for culturing <aerobic or facultative anaerobic microorganisms, etc.>. Specifically, it can be applied not only to the production of foods, chemicals, chemicals, etc. by brewing, fermentation, etc., the production of reaction products by microorganisms, such as the production of bioethanol using biomass, but also to the growth of microorganisms, etc. .. When culturing <aerobic or facultative anaerobic microorganisms, etc.>, a gas containing oxygen is used as a useful gas.

In the biological reaction using <aerobic or facultative anaerobic microorganisms, etc.>, the reaction products may be produced by the microorganisms, etc. using the culture solution as a nutrient source in the culture solution containing the microorganisms, etc. contained in the culture tank. , Microorganisms, etc. are propagated.

As the culture solution of <aerobic or facultative anaerobic microorganisms, etc.>, one containing saccharides and nitrogen sources is used. As the saccharide, maltose, sucrose, glucose, fructose, saccharides such as mixtures thereof, ethanol and the like are usually used, and the concentration of the saccharide in the culture solution is not particularly limited, but is 0.1 to 10 w / v%. Is preferable. As the nitrogen source, ammonium chloride, ammonium sulfate or corn steep liquor, yeast extract, meat extract, peptone and the like are used, and the nitrogen source is preferably 0.1 to 10 w / v%. Further, it is preferable to add vitamins, inorganic salts and the like to the culture broth, if necessary, in addition to the sugar and nitrogen sources.

<Aerobic or facultative anaerobic microorganisms, etc.> include aerobic and facultative bacteria such as aspergillus, natto, acetic acid, yeast, and lactic acid, which are conventionally used in technical fields such as brewing and fermentation. In addition to sexually anaerobic microorganisms, various aerobic and facultative anaerobic microorganisms created by gene recombination technology can be used. In addition, examples of the cells include animal cells for producing physiologically active peptides or proteins used as antibody drugs, particularly recombinant animal cells and the like.

Furthermore, the biological reaction apparatus / method of the present invention can also be used for culturing <obligately anaerobic microorganisms, etc.> whose growth is inhibited by exposure to oxygen having an atmospheric level concentration such as bifidobacteria. When culturing <obligately anaerobic microorganisms, etc.>, nitrogen is used as a useful gas.

Further, the biological reaction apparatus / method of the present invention produces organic substances such as amino acids, organic acids and proteins from carbon gas (carbon dioxide gas, methane gas) derived from natural gas, carbon dioxide gas discharged from a thermal power plant, etc. < It can also be used for culturing organic matter synthetic microorganisms and the like. When culturing <organic synthetic microorganisms, etc.>, carbon gas derived from natural gas, carbon dioxide gas discharged from a thermal power plant, or the like is used as a useful gas.

<Micro bubbles used in the biological reaction apparatus / method of the present invention>
Next, the fine bubbles used in the biological reaction apparatus / method of the present invention will be described.

The "fine bubble / ultra fine bubble" (fine bubble) used in the biological reaction apparatus / method of the present invention means "fine bubble" and / or "ultra fine bubble". While "ordinary bubbles" rapidly rise in water and burst at the surface and disappear, "fine bubbles", which are micro bubbles with a diameter of 50 μm or less, shrink and disappear in water, and at this time, they disappear. Along with free radicals, "ultra fine bubbles", which are ultrafine bubbles having a diameter of 100 nm or less, are generated, and these "ultra fine bubbles" remain in water for a relatively long time.

In the present invention, bubbles having a number average diameter of 100 μm or less are referred to as “fine bubbles”, and bubbles having a number average diameter of 1 μm or less are referred to as “ultra fine bubbles”. Image analysis method, laser diffraction scattering method, electrical detection band method, resonance type mass measurement method, optical fiber probe method, etc. are generally used as methods for measuring the bubble diameter of "fine bubbles / ultra fine bubbles". As a method for measuring the bubble diameter of nanobubbles, a dynamic light scattering method, a brown motion tracking method, an electrical detection band method, a resonance type mass measurement method and the like are generally used.

<Characteristics of the biological reaction apparatus of the present invention>
First, a conventionally used biological reaction device for culturing microorganisms and the like containing fine bubbles of an oxygen-containing gas will be described.

As shown in FIG. 12, in the conventional biological reaction device, the biological culture solution 103 is extracted from the culture tank 102 by the culture tank pump 101, and the oxygen-containing gas a is supplied by the fine bubble generator 104 to which the oxygen-containing gas a is supplied. The biological culture solution 103 in the culture tank 102 is stirred by the stirrer 105 while recirculating the culture tank 102 to contain the fine bubbles of the above. Further, a filter (not shown) is arranged between the culture tank pump 101 and the fine bubble generator 104, and the filtrate separated from the biological culture solution 103 extracted from the culture tank 102 is used as the fine bubble generator 104. It is also supplied to.

As shown in [FIG. 1], the biological reaction apparatus of the present invention has a culture tank 2 for accommodating the biological culture solution 1, an extraction pipeline 4 for extracting the biological culture solution 1 from the culture tank 2 by a culture tank pump 3 or the like. A biological reaction apparatus provided between the extraction conduit 4 and the culture tank 2 and provided with a multi-nozzle 5 for returning the biological culture solution 1 to the culture tank 2.
The multi-nozzle 5 includes a fine bubble generator 5-1 for containing fine bubbles of gas A containing a useful gas in the biological culture solution 1, and a plurality of discharge ports 5-2 provided at outlets on the culture tank 2 side. Equipped with
The plurality of discharge ports 5-2 are provided so as to discharge the microbubble-containing biological culture solution in multiple directions of 0 ° to 90 ° with respect to the front direction of the central axis B of the multi-nozzle 5. It is a feature.

In the first embodiment of the biological reaction device / method of the present invention shown in [FIG. 1], the multi-nozzle 5 is provided at the bottom of the culture tank 2, but the biological reaction device / device of the present invention shown in [FIG. 2]. As in the second embodiment of the method, the multi-nozzle 5 may be provided on the side surface of the culture tank 2.

As described above, when the fine bubble-containing biological culture solution of the gas A containing the useful gas is returned to the culture tank 2, the temperature is 0 ° to 90 ° with respect to the front direction of the central axis B of the multi-nozzle 5. By discharging in the direction, it is possible to provide a biological reaction apparatus and a biological reaction method capable of maintaining the activity of microorganisms and the like without using a stirrer. By not using the stirrer, a) the microorganisms are stressed and damaged and the activity of the microorganisms is reduced, and b) the part where sterility is difficult to be performed before the biological reaction (the seal part of the rotating shaft of the stirrer, rotation). Shaft bearings, stirring blades, baffles, baffles, etc.) are generated and it is difficult to prevent contamination, c) Installation, operation, maintenance, management, etc. of the stirring machine are costly. You can solve the problem. Furthermore, since it is not necessary to provide members such as baffles and baffles on the inner surface of the culture tank, Teflon processing (Teflon: registered trademark) is applied to the inner surface of the culture tank to prevent dirt from adhering and to prevent contamination. Risk can be reduced.

In the biological reaction apparatus of the present invention, a filtration that separates the biological culture solution 1 extracted from the culture tank 2 into a filtrate and a biological culture solution excluding the filtrate between the extraction conduit 4 and the multi-nozzle 5. A vessel can be arranged and this filtrate can be supplied to the multi-nozzle 5. In this way, the stress and damage received by the microorganisms and the like are reduced by containing the fine bubbles of the gas A containing the useful gas in the filtrate excluding the microorganisms and discharging the fine bubbles from the discharge port 5-2 of the multi-nozzle 5. can do. The biological culture solution excluding the filtrate is collected or returned to the culture tank 2 through another pipe.

<Multi-nozzle used in the biological reaction apparatus of the present invention>
Next, a preferred embodiment of the multi-nozzle used in the biological reaction apparatus of the present invention will be described with reference to FIGS. 3 to 5.

As a suitable multi-nozzle 5, as shown in [FIG. 3],
1) Lower end 5-3 connected to the extraction pipe 4,
2) Upper end 5-4 connected to the culture tank 2,
3) A plurality of transport pipelines 5-5 provided along the central axis B of the nozzle, which divide the biological culture solution 1 supplied to the lower end portion 5-3 and transport it to the upper end portion 5-4 side.
4) A plurality of discharge pipes 5-6 connected to each transport pipe 5-5, and 5) a plurality of discharge ports 5 provided at outlets on the culture tank 2 side of each discharge pipe 5-6. -2
The plurality of discharge ports 5-2 are provided so as to discharge the fine bubble-containing biological culture solution in multiple directions of 0 ° to 90 ° with respect to the culture tank 2 side direction of the central axis B. It is characterized by being.

As described above, the plurality of discharge ports 5-2 of the multi-nozzle 5 are usually multi-directional in which the microbubble-containing biological culture solution is 0 ° to 90 ° with respect to the culture tank 2 side direction of the central axis B. However, when a plurality of discharge ports 5-2 are installed so as to project inward from the wall surface of the culture tank 2, the microbubble-containing biological culture solution is placed on the culture tank 2 side of the central axis B. It is also possible to discharge in a direction that exceeds 90 ° and is less than 180 ° with respect to the direction. In this way, when the microbubble-containing biological culture solution is discharged in the direction of more than 90 ° and less than 180 ° with respect to the culture tank 2 side direction of the central axis B, the fine bubble-containing biological culture solution is formed on the inner wall surface of the culture tank 2. Since there are actions such as collision with each other and suppression of floating of fine bubbles, it is possible to increase options when designing for appropriately stirring the biological culture solution 1 contained in the culture tank 2.

[FIG. 4] is a schematic cross-sectional view showing the XX'cross section of FIG. 3. As described above, the plurality of transport pipelines 5-5 are provided along the central axis B of the multi-nozzle. Further, [FIG. 5] is an external schematic view showing the left end surface of [FIG. 3]. As described above, the plurality of discharge ports 5-2 are symmetrical with respect to the central axis B of the multi-nozzle. It is preferably arranged along the periphery of the upper end portion 5-4.

[FIG. 3] does not show the fine bubble generator 5-1. However, as an aspect of installing the fine bubble generator 5-1
a) A mode in which one fine bubble generator 5-1 is provided between the lower end portion 5-3 and the transport line 5-5.
b) A mode in which a fine bubble generator 5-1 is provided in each discharge pipe line 5-6, or a mode in which a fine bubble generator 5-1 is provided in each transport pipe line 5-5 is adopted. Can be done.

The aspect of a) above is preferable from the viewpoint of cost of installation, maintenance, etc., and the aspects of b) and c) above individually adjust the amount of fine bubbles of the biological culture solution 1 discharged from each discharge port 5-2. It is preferable from the viewpoint of being able to do it.

<Discharge of biological culture solution containing fine bubbles>
In the multi-nozzle of the biological reaction device of the present invention, the plurality of discharge ports discharge the fine bubble-containing biological culture solution in multiple directions of 0 ° to 90 ° with respect to the front direction of the central axis of the multi-nozzle. It is provided. As a result, the biological culture solution contained in the culture tank can be sufficiently stirred by the fine bubble-containing biological culture solution discharged from the plurality of discharge ports without using a stirrer, and the activity of microorganisms and the like can be maintained. Can be done.

A pair of transport pipes 5-5 and discharge pipes connected in order to discharge the fine bubble-containing biological culture solution in multiple directions of 0 ° to 90 ° with respect to the front direction of the central axis B of the multi-nozzle. The positional relationship of 5-6 can be set as follows.

[Position A]
As shown in FIGS. 6 and 7, in each plane C including a pair of connected transport pipes 5-5 and discharge pipe 5-6, the transport pipe 5-5 and the discharge pipe 5 are used. Set the angle α on the acute angle side formed by -6 to more than 90 ° and less than 180 °.

Due to this positional relationship A, the direction of the microbubble-containing biological culture solution discharged from each discharge port 5-2 is set to multiple directions away from the central axis B of the multi-nozzle, and the biological culture solution 1 housed in the culture tank 2 is set. Can be agitated.

Assuming that the number of discharge ports 5-2 is n, the angles α1 to αn at each pair 1 to n of the connected transport line 5-5 and the discharge line 5-6 are more than 90 ° and less than 180 °. It can be set individually in the range as appropriate.

[Position B]
As shown in FIG. 7, each plane C including a pair of connected transport pipes 5-5 and discharge pipe 5-6, and the central axis B of the transport pipe and the multi-nozzle included in each plane C. The angle β on the acute angle side formed by the plane D including the above is set to be more than 0 ° and less than 90 °.

Due to this positional relationship B, the biological culture solution 1 housed in the culture tank 2 is provided with the direction of the fine bubble-containing biological culture solution discharged from each discharge port 5-2 as a multi-direction centered on the central axis B. Can be agitated.

Assuming that the number of discharge ports 5-2 is n, there are n pairs of planes C and corresponding planes D, but the angles β1 to βn in each pair are individually in the range of more than 0 ° and less than 90 °. It can be set as appropriate.

The angles α1 to αn and the angles β1 to βn allow the biological culture solution 1 contained in the culture tank 2 to be appropriately agitated according to the shape of the culture tank 2, the number of discharge ports 5-2, the installation position, and the like. In addition, it can be set appropriately by experiments, simulations, and the like.

<Shape and structure of culture tank>
As the shape of the culture tank used in the biological reaction apparatus / method of the present invention, generally used cylindrical, cubic, or rectangular parallelepiped shapes can be used, but from the viewpoint of uniform and uniform stirring, they can be used. , Cylindrical one is preferable.

Further, as the culture tank used in the biological reaction apparatus / method of the present invention, the first embodiment of the biological reaction apparatus / method of the present invention shown in [FIG. 1] and the biological reaction apparatus of the present invention shown in [FIG. 2] are used. -A vertical culture tank as in the second embodiment of the method or a horizontal culture tank as in the third embodiment of the biological reaction apparatus / method of the present invention shown in [FIG. 8] can be used, but the horizontal type. It is preferable to use a culture tank. By using the horizontal culture tank as shown in [FIG. 8], the number of multi-nozzles and the number of multi-nozzles can be determined by experiments, simulations, etc. so that the biological culture solution 1 contained in the culture tank 2 can be appropriately stirred. It is possible to increase the choices when designing the installation position, etc., and it becomes easier to realize the desired stirring state. Further, assuming that the amount of fine bubbles supplied per unit volume of the biological culture solution 1 is constant, the amount of fine bubbles per surface of the biological culture solution 1 can be reduced when the horizontal culture tank is used. The thickness of the bubbles formed on the surface of the biological culture solution 1 can be reduced.

<Number of multi-nozzles installed>
In the biological reaction apparatus / method of the present invention, it is preferable that a plurality of multi-nozzles are provided. By providing a plurality of multi-nozzles, the number of multi-nozzles, the installation position of the multi-nozzles, etc. are designed by experiments, simulations, etc. so that the biological culture solution 1 contained in the culture tank 2 can be appropriately agitated. It is possible to increase the choices at the time, and it becomes easier to realize the desired stirring state.

Further, when designing to realize a desirable stirring state by independently adjusting the discharge amount of the fine bubble-containing biological culture solution discharged from the plurality of multi-nozzles in each of the plurality of multi-nozzles. You can further increase the choices of.

<Fine bubble generator installed in the multi-nozzle>
As the fine bubble generator provided in the multi-nozzle, a known or commercially available water flow type fine bubble generator can be used.

Examples of the water flow type fine bubble generator include those shown in [Fig. 9]. In this fine bubble generator 6, the biological culture solution is supplied from the inlet portion 6-1 of the fine bubble generator 6 under pressure, and the diameter of the conduit is narrowed to increase the flow velocity, while the throat portion 6-2. Generates turbulence. In this state, the gas A containing the useful gas is supplied from the gas inlet 6-3, mixed with the biological culture solution in the suction part 6-4, becomes fine bubbles by the water flow, and the useful gas is introduced from the outlet part 6-5. The biological culture solution containing the fine bubbles of the contained gas A is discharged.

Further, preferably, gas inlets 6-3 as shown in [FIG. 10] to [FIG. 11] are continuously provided on the side surfaces along the plane perpendicular to the central axis of the fine bubble generator 6. A fine bubble generator with a slit can be used. When such a fine bubble generator is used, as shown in [FIG. 11], the gas A containing the useful gas blown from the slit has a wide continuous gas along the inner surface of the fine bubble generator 6. The thin layer E is formed and flows along the inner surface of the fine bubble generator 6, and the fine bubbles F are gradually formed, and a large amount of fine bubbles F are formed near the outlet portion 6-5, so that the fine bubbles F are formed. It is possible to improve the production efficiency of the gas and efficiently and sufficiently increase the fine bubble content of the biological culture solution. Further, since the thin layer F acts as a cushion for preventing microorganisms and the like from colliding with the inner surface of the fine bubble generator 6, it is possible to reduce stress and damage to the microorganisms and the like.

<Others>
The upper limit of the oxygen content of the fine bubbles is less than 60%, preferably 55% or less, more preferably 50% or less, and most preferably 45% or less. If the oxygen content of the fine bubbles is excessively increased to 60% or more, the stress and damage to the microorganisms due to the oxidizing action of oxygen will increase. The lower limit of the oxygen content of the fine bubbles is 23% or more, preferably 25% or more, more preferably 27% or more, and most preferably 30% or more. If the oxygen content of the fine bubbles is excessively small, less than 23%, the dissolved oxygen concentration decreases, and it becomes difficult to increase the activity of microorganisms such as microorganisms. In order to obtain a gas that forms fine bubbles with an increased oxygen content, usually, a PSA method using an adsorbent, a VSA method, etc., an electrolysis method for water, a cold separation method, a membrane separation method, a chemical adsorption method, etc. It is preferable to increase the oxygen content of the gas by using a known oxygen enrichment means such as, and from an economical point of view, it is preferable to use an oxygen enrichment membrane.

Further, as the culture tank pump 3 for extracting the biological culture solution 1 from the culture tank 2, a positive displacement pump such as a tube pump, a diaphragm pump, a screw pump, a rotary pump, etc., which has relatively little stress and damage to microorganisms and the like, is suitable. Can be used.

<Summary>
As described above, in the biological reaction apparatus / method of the present invention, a biological culture solution containing fine bubbles of a gas containing a useful gas is discharged in multiple directions in a culture tank using a multi-nozzle. By doing so, the biological culture solution contained in the culture tank can be sufficiently stirred without using a stirrer, and the activity of microorganisms and the like can be maintained.

Further, in the biological reaction apparatus / method of the present invention, by not using a stirrer, the internal structure of the culture tank / biological reaction apparatus can be simplified, the detergency can be improved, and the contamination by various germs can be prevented.

By not using the stirrer, a) the microorganisms are stressed and damaged and the activity of the microorganisms is reduced, and b) the part where sterility is difficult to be performed before the biological reaction (the seal part of the rotating shaft of the stirrer, rotation). Shaft bearings, stirring blades, baffles, baffles, etc.) are generated, making it difficult to prevent contamination, and c) costly installation, operation, maintenance, and management of the stirring machine. Can be resolved.

As described above, by using the multi-nozzle without using the stirrer, the biological culture solution contained in the culture tank can be agitated economically and efficiently.

1 Biological culture solution (containing culture solution, microorganisms, etc.) 2 Culture tank 3 Culture tank pump 4 Extraction pipeline 5 Multi-nozzle 5-1 Fine bubble / ultra fine bubble generator 5-2 (Multiple) ) Discharge port 5-3 Lower end 5-4 Upper end 5-5 (Multiple) transport pipelines 5-6 (Multiple) Discharge pipelines 6 Fine bubble generator 6-1 Inlet 6-2 Throat 6- 3 Gas inlet 6-4 Suction part 6-5 Outlet part A Gas containing useful gas B (multi-nozzle) central axis C Each plane including a pair of connected transport pipes and discharge pipes D In each plane C Plane E including the transport pipeline included and the central axis B of the multi-nozzle Wide thin layer F Fine bubbles α In each plane C, the angle β on the sharp angle side formed by the transport pipeline and the discharge pipeline β Each plane C and each plane Angle on the sharp side formed by the plane D including the transport pipeline included in C and the central axis B of the multi-nozzle 101 Culture tank pump 102 Culture tank 103 Biological culture liquid 104 Fine bubble generator 105 Stirrer a Oxygen-containing gas

Claims (13)

A culture tank containing a culture medium and a biological culture medium containing microorganisms or cells,
A biological reaction apparatus provided with an extraction pipe for extracting the biological culture solution from the culture tank, and a multi-nozzle arranged between the extraction pipe and the culture tank and returning the biological culture solution to the culture tank. And
The above multi-nozzle
A fine bubble / ultra fine bubble generator for containing a gas fine bubble / ultra fine bubble containing a gas useful for culturing the microorganism or cell in the biological culture solution, and an outlet on the culture tank side. Equipped with multiple discharge ports,
The plurality of discharge ports discharge the biological culture solution containing the fine bubble / ultra fine bubble in multiple directions of 0 ° to 90 ° with respect to the culture tank side direction of the central axis of the multi-nozzle. A biological reaction device, characterized in that it is provided in such a manner. A filter for separating the biological culture solution extracted from the culture tank into the filtrate and the biological culture solution excluding the filtrate is arranged between the extraction conduit and the multi-nozzle.
The biological reaction apparatus according to claim 1, wherein the filtrate is supplied to the multi-nozzle. The above multi-nozzle
The lower end connected to the above extraction pipe,
The upper end connected to the above culture tank,
A plurality of transport pipelines provided along the central axis of the multi-nozzle, which are connected to the lower end portion and which divide the biological culture solution supplied to the lower end portion and transport the biological culture solution to the upper end portion side.
It is provided with a plurality of discharge pipes connected to each of the transport pipes, and the plurality of discharge ports provided at the outlets of the discharge pipes on the culture tank side.
In each plane including the pair of the transfer pipes and the discharge pipes connected to each other, the angle on the acute angle side formed by the transport pipes and the discharge pipes is set to be more than 90 ° and less than 180 °. The biological reaction apparatus according to claim 1 or 2, wherein the biological reaction apparatus is characterized. The biological reaction apparatus according to claim 3, wherein each plane of the multi-nozzle is a plane including a central axis of the multi-nozzle. In the multi-nozzle, each of the planes is a plane that does not include the central axis of the multi-nozzle, and the angle on the acute angle side formed by each plane and the plane including each transport pipeline and the central axis of the multi-nozzle is 0. The biological reaction apparatus according to claim 3, wherein the temperature is set to more than ° and less than 90 °. The biological reaction apparatus according to any one of claims 3 to 5, wherein in the multi-nozzle, each of the discharge pipes is provided with the fine bubble / ultra fine bubble generator. The biological reaction according to any one of claims 3 to 5, wherein in the multi-nozzle, one fine bubble / ultra fine bubble generator is provided between the lower end portion and the transport pipeline. Device. The biological reaction apparatus according to any one of claims 3 to 5, wherein in the multi-nozzle, the fine bubble / ultra fine bubble generator is provided in each of the transport pipelines. The biological reaction apparatus according to any one of claims 1 to 8, wherein the culture tank is a horizontal culture tank. The biological reaction apparatus according to any one of claims 1 to 8, wherein the culture tank is a vertical culture tank. The biological reaction apparatus according to any one of claims 1 to 10, wherein a plurality of the multi-nozzles are provided. It is characterized in that the discharge amount of the biological culture solution containing the fine bubble / ultra fine bubble discharged from the plurality of multi-nozzles is independently adjusted in each of the plurality of multi-nozzles. The biological reaction apparatus according to claim 11. The biological reaction method according to any one of claims 1 to 12, wherein the reaction product of the microorganism or cell is obtained, or the microorganism or cell is grown.

Images