WO2017017830A1 - Bioreactor using oxygen-enriched micro/nano-bubbles, and bioreaction method using bioreactor using oxygen-enriched micro/nano-bubbles - Google Patents

Abstract

This bioreactor and the bioreaction method that uses the bioreactor address the problems of reducing the stress/damage incurred by a microorganism or the like during a bioreaction and of making it possible to efficiently and economically perform a bioreaction that uses a microorganism or the like. In order to solve said problems, this bioreactor and the bioreaction method that uses the bioreactor are characterized in that a micro/nano-bubble generator is used to incorporate oxygen-enriched micro/nano-bubbles (MNB), which are formed from air that has an increased concentration of oxygen, into a biological culture solution that has been extracted from a culture tank and in that the biological culture solution into which the oxygen-enriched MNB have been incorporated is returned to the culture tank. Either 1) a method wherein the biological culture solution that has been extracted from the culture tank is separated by means of a filter into a filtrate and into a biological culture solution from which the filtrate has been removed and the oxygen-enriched MNB are incorporated into the filtrate or 2) a method wherein the oxygen-enriched MNB are directly incorporated into the biological culture solution that has been extracted from the culture tank may be employed as a method for incorporating the oxygen-enriched MNB into the biological culture solution that has been extracted from the culture tank.

Description

Biological reaction device using oxygen-enriched micro-nano bubbles and biological reaction method using this biological reaction device

The present invention cultivates microorganisms or cells (hereinafter referred to as “microorganisms”) to produce reaction products in the microorganisms or the like, and to propagate microorganisms and the like, and organisms using this biological reaction apparatus Regarding the reaction method, a micro-nano bubble formed from air with an increased oxygen concentration (hereinafter referred to as “micro-nano bubble” as “MNB”, “nano bubble” as “NB”, It is characterized in that a biological reaction is efficiently performed by containing “micro-nano bubbles formed from elevated air” in some cases “oxygen-enriched MNB”.

Unlike chemical reactions, biological reactions are slow, but they do not use a lot of energy or chemicals, so they are mild and meaningful for the environment.
However, the biological reaction generally has a problem that the reaction is mild and slow. That is, for chemical reactions, a reaction within one hour is often sufficient, whereas in the case of biological reactions, a reaction time of several days or longer, several days or particularly several weeks or longer is required. There is also. For this reason, it is required to perform biological reactions efficiently and economically.

As techniques for improving the efficiency of biological reactions, Patent Documents 1 to 3 promote the activation of microorganisms and the like by causing the presence of MNB or NB formed from air in the culture medium in the culture of microorganisms and the like. In addition, it is disclosed that the reaction efficiency of biological reactions, reduction of reaction time, and the like are attempted.

Specifically, Patent Document 1 describes that air MNB and NB are mixed in the culture solution before the culture solution is supplied to the culture tank. It is described that air MNB is mixed before the liquid is supplied to the culture vessel. Further, in Patent Document 3, in a batch system, a culture solution is extracted from a culture tank, filtered through a bacterial cell filter to obtain a filtrate, and this filtrate is mixed with air MNB and refluxed to the culture tank. It is described.

However, in an apparatus in which the culture solution supplied to the culture vessel contains air MNB and NB as disclosed in Patent Documents 1 and 2, an appropriate amount of the culture solution in the culture vessel is appropriate in the initial stage of the biological reaction. Although the content of MNB and / or NB in the culture medium in the culture tank cannot be properly maintained over the entire long-term biological reaction, the reaction of the biological reaction Efficiency, shortening of reaction time, etc. cannot be achieved sufficiently.

Moreover, in the said patent document 3, as shown in FIG. 7, a culture solution is extracted from the culture tank 107 as a biological reaction tank, and it filters with the microbial cell filter 110, A filtrate is obtained, Micro-nano bubble generation | occurrence | production in this filtrate In the tank 115, an apparatus for generating and mixing air MNB by the micro / nano bubble generator 116 and returning it to the culture tank is described. In this apparatus, the MNB content of the culture solution in the culture tank is set to an appropriate value. Although it can be maintained, microorganisms and the like are subjected to stress and damage in the step of extracting the culture solution from the culture vessel, the step of filtering the culture solution with a cell strain filter, the step of returning the culture solution excluding the filtrate to the culture vessel, etc. Therefore, there exists a problem that the activity of microorganisms will fall.

Therefore, the present inventors made the oxygen concentration of the gas constituting MNB higher than the concentration in air (about 21%),
-Even if the amount of the biological culture solution extracted from the culture tank is reduced and the amount of MNB contained in the biological culture solution in the culture tank is reduced, high concentration oxygen that is easily absorbed can be supplied to microorganisms. Therefore, the activity of microorganisms etc. can be maintained,
〇 By reducing the amount of the biological culture solution withdrawn from the culture tank, stress and damage to microorganisms can be reduced and the energy required for circulation of the biological culture solution can be reduced.
〇 By reducing the amount of MNB contained in the biological culture, the energy required for driving the MNB generator can be reduced.
It has been found that there is a great merit, and the present invention has been achieved.

In addition, diaphragm pumps, tube pumps, screw pumps with relatively little stress and damage to microorganisms, etc. as a pump for circulating the biological culture solution to the outside of the culture vessel as the amount of biological culture solution extracted from the culture vessel decreases. It has also been found that a positive displacement pump such as a rotary pump can be suitably used, and this can further reduce stress and damage to microorganisms.

Japanese Patent No. 4805120 Japanese Patent No. 4956552 Japanese Patent No. 4146476

The problem of the biological reaction device of the present invention and the biological reaction method using this biological reaction device is to reduce the stress and damage to the microorganisms during the biological reaction, and the biological reaction using the microorganisms is efficient and economical. It is to be able to do it.

In order to solve the above-mentioned problems, the biological reaction apparatus of the present invention and the biological reaction method using this biological reaction apparatus are characterized in that oxygen-enriched MNB is contained in a biological culture solution containing microorganisms and the like. is there.

In addition, as a pump for circulating the biological culture solution outside the culture tank, by suitably using a positive displacement pump such as a diaphragm pump, a tube pump, a screw pump, or a rotary pump that has relatively little stress and damage to microorganisms, etc. The problem can be further solved.

In the present invention, the oxygen concentration of the gas constituting the MNB is made higher than the concentration in air (about 21%), thereby reducing the amount of the biological culture solution extracted from the culture tank, and the MNB contained in the biological culture solution. Even if the amount is reduced, it is possible to supply a high concentration of easily absorbed oxygen in the MNB state to the microorganism or the like, and to maintain the activity of the microorganism or the like.
Furthermore, by reducing the amount of the biological culture solution extracted from the culture tank, it is possible to reduce stress and damage to microorganisms and the like, and to reduce the energy required for circulation of the biological culture solution.
Furthermore, by reducing the amount of MNB contained in the culture solution, the energy required for driving the MNB generator can be reduced.

In addition, diaphragm pumps, tube pumps, screw pumps with relatively little stress and damage to microorganisms, etc. as a pump for circulating the biological culture solution to the outside of the culture vessel as the amount of biological culture solution extracted from the culture vessel decreases. Further, a positive displacement pump such as a rotary pump can be suitably used, and this can further reduce stress and damage to microorganisms.
As described above, the present invention is excellent in that a biological reaction using microorganisms or the like can be performed efficiently and economically.

It is a schematic diagram which shows 1st Embodiment of the biological reaction apparatus of this invention. It is sectional drawing which shows the outline | summary of the nozzle type micro nano bubble generator used in 1st Embodiment. It is sectional drawing which shows the outline | summary of the oxygen enrichment means used in 1st Embodiment. It is a schematic diagram which shows 2nd Embodiment of the biological reaction apparatus of this invention. It is a schematic diagram which shows 3rd Embodiment of the biological reaction apparatus of this invention. It is a schematic diagram which shows 4th Embodiment of the biological reaction apparatus of this invention. It is FIG. 1 of patent document 3 (patent 4146476 gazette) which is a prior art example.

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.

First, general items of the biological reaction apparatus and biological reaction method of the present invention will be described.

The bioreactor of the present invention and the bioreaction method using the bioreactor include reaction products produced by microorganisms such as brewing, fermentation, etc. for the production of foods, chemicals, chemicals, etc., and the production of bioethanol using biomass. It is useful not only for the production of but also for the growth of microorganisms and the like.

The biological reaction of the present invention is to cause a reaction product to be produced by a microorganism or the like in a culture solution containing a microorganism or the like contained in a culture tank, and to cause the microorganism to grow.

As the culture solution in the present invention, a culture solution containing a saccharide and a nitrogen source is used. As saccharides, saccharides such as maltose, sucrose, glucose, fructose, and mixtures thereof are usually used. The concentration of saccharides in the culture solution is not particularly limited, but is set to 0.1 to 10 w / v%. preferable. As the nitrogen source, ammonium chloride, ammonium sulfate, corn steep liquor, yeast extract, meat extract, peptone or the like is used, and it is preferably set to 0.1 to 10 w / v%. Furthermore, it is preferable to add vitamins, inorganic salts, and the like to the culture solution as needed in addition to the saccharides and the nitrogen source.

Examples of the microorganism in the present invention include aerobic and facultative anaerobic microorganisms such as koji molds such as Aspergillus, Bacillus natto, acetic acid bacteria, yeasts, and lactic acid bacteria conventionally used in technical fields such as brewing and fermentation. Various aerobic and facultative anaerobic microorganisms created by gene recombination technology can be used. Examples of the cells include animal cells for producing physiologically active peptides or proteins used as antibody drugs, particularly genetically modified animal cells.
The concentration of the microorganism or cell added to the culture solution is not particularly limited, but is preferably 0.5 to 10 g / L, and more preferably 3.0 to 6.0 g / L.

Next, features of the biological reaction apparatus and biological reaction method of the present invention will be described.
As described above, the first feature of the present invention is that the biological culture liquid extracted from the culture tank contains oxygen-enriched MNB, and the biological culture liquid containing the oxygen-enriched MNB is added to the culture tank. To reflux.

Thus, by making the oxygen concentration of the gas constituting the MNB higher than the concentration in air (about 21%), the amount of the biological culture solution extracted from the culture tank is reduced, and the MNB contained in the biological culture solution is contained. Even if the amount is reduced, it is possible to supply high-concentration oxygen in the MNB state that is easily absorbed to the microorganisms in the culture tank, and the activity of the microorganisms can be maintained.

Furthermore, by reducing the amount of the biological culture solution extracted from the culture tank, it is possible to reduce stress and damage to microorganisms and the like, and to reduce the energy required for circulation of the biological culture solution.
Furthermore, by reducing the amount of MNB contained in the culture solution, the energy required for driving the MNB generator can be reduced.

The second feature of the present invention is that, as described above, by using oxygen-enriched MNB, the amount of the biological culture solution extracted from the culture vessel can be reduced, and the biological culture solution is circulated outside the culture vessel. As a pump to be used, a positive displacement pump such as a diaphragm pump, a tube pump, a screw pump, a rotary pump, or the like that causes relatively little stress and damage to microorganisms or the like is preferably used.
The use of such a positive displacement pump can further reduce stress and damage to microorganisms.

The “micro nano bubble” of the present invention means “micro bubble” and / or “nano bubble”. While “normal bubbles” rapidly rise in water and burst and disappear on the surface, microbubbles with a diameter of 50 μm or less called “microbubbles” shrink in water and disappear. Together with free radicals, “nanobubbles”, which are ultrafine bubbles having a diameter of 100 nm or less, are generated, and these “nanobubbles” remain in water for a certain amount of time.

“Nano bubbles”, which are very small bubbles, are also called “ultra fine bubbles”. Currently, in the ISO (International Organization for Standardization), the creation of an international standard for fine bubble technology is being considered, and once the international standard is created, the name of “nanobubble”, which is currently commonly used, There is a possibility that it will be unified into “Ultra Fine Bubble”.

As the micro / nano bubble generating device, a known or commercially available device can be used. For example, after a sufficient amount of gas is dissolved in water at a certain high pressure, the dissolved gas is released by releasing the pressure. “Pressure-dissolving microbubble generator” that creates supersaturation conditions, utilizing the phenomenon that bubbles are broken apart when large bubbles are entrained in the vortex by causing a water flow and large vortices are engulfed in the vortex The “gas-liquid two-phase flow swirl type microbubble generator” or the like can be used.

Examples of nanobubble generators include, for example, JP 2007-31690 A, JP 2006-289183 A, JP 2005-245817 A, JP 2007-136255 A, and JP 2009-39600 A. Those described can be used.
As the micro / nano bubble generating device, it is preferable to use a device driven by a water flow (nozzle method) because a large amount of MNB can be generated economically.

In order to obtain air with an increased oxygen concentration, known oxygen-enriching means such as PSA method using adsorbent, VSA method, water electrolysis method, cryogenic separation method, membrane separation method, chemical adsorption method, etc. However, it is preferable to use an oxygen-enriched film from an economical viewpoint.

The oxygen concentration of the oxygen-enriched MNB is preferably 25 to 40%, more preferably 35 to 40%. When the oxygen concentration is 25% or more, the respiratory action of microorganisms and the like can be promoted, and the activity of microorganisms and the like can be increased. When the oxygen concentration is 40% or less, microorganisms and the like are not easily damaged by oxidation.

As described above, the first feature of the present invention is that the biological culture liquid extracted from the culture tank contains oxygen-enriched MNB, and the biological culture liquid containing the oxygen-enriched MNB is added to the culture tank. The following two methods can be adopted as a method of adding oxygen-enriched MNB to the biological culture solution extracted from the culture tank.
1) A method in which a biological culture liquid extracted from a culture tank is separated into a biological culture liquid from which the filtrate and the filtrate are removed by a filter, and this filtrate contains oxygen-enriched MNB.
2) A method of directly adding oxygen-enriched MNB to a biological culture solution extracted from a culture tank.

In the above method 1), oxygen-enriched MNB is blown into the filtrate that does not substantially contain microorganisms. Therefore, the microorganisms are not subjected to stress or damage in the step of blowing oxygen-enriched MNB. In some cases, stress and damage may occur during the filtration process. In addition, since the amount of the filtrate separated in the filtration step is small (the amount of the filtrate is usually about 1/10 to 1/100 of the amount of the biological culture liquid extracted from the culture tank), In order to supply a sufficient amount of oxygen-enriched MNB to the biological culture solution, it may be necessary to increase the amount of the biological culture solution extracted from the culture tank, or to increase the amount of oxygen-enriched MNB blown in, There is a possibility that the operating cost of the apparatus becomes high and the stress and damage to which microorganisms are subjected are also increased.

In the method 2), oxygen-enriched MNB is blown into the biological culture solution extracted from the culture tank containing microorganisms, so that the microorganisms are stress-damaged in the step of blowing oxygen-enriched MNB. In some cases, as in the above method 1), stress damage may be reduced in the filtration step. Further, in the above method 1), even when it is necessary to increase the amount of oxygen-enriched MNB, if the above method 2) is used, the oxygen-enriched MNB is directly added to the biological culture solution extracted from the culture tank. Therefore, it is not necessary to increase the amount of the biological culture solution, and the operating cost of the apparatus is not increased, and the stress and damage to the microorganisms are not increased.

The biological reaction apparatus employing the method 1) will be described based on the first embodiment of the present invention shown in FIG. 1, and the biological reaction apparatus employing the method 2) will be illustrated in FIG. A description will be given based on the second embodiment of the invention.

○ First embodiment (FIG. 1)
First, a first embodiment of the present invention will be described with reference to FIG.
The first embodiment is a biological reaction apparatus for causing a microorganism or the like to generate a reaction product, and includes oxygen-enriched MNB in a biological culture solution as follows.
a) The culture solution 1 is supplied to the culture tank 2.
b) The valve 12 is closed, the valve 13 and the valve 14 are opened, the culture tank pump 8 is driven, and the biological culture solution 3 containing the culture solution, microorganisms and the like is extracted from the culture vessel 2 and supplied to the filter 4.
c) The biological culture solution B (that is, the biological culture solution in which microorganisms and the like are concentrated) separated by the filter 4 and excluding the filtrate is returned to the culture tank 2.
d) The filtrate A separated by the filter 4 is stored in the micro / nano bubble generating tank 6, and oxygen enriched MNB is contained by the micro / nano bubble generating device 7a.
g) The return pump 9 is driven to return the filtrate D containing oxygen-enriched MNB to the culture tank 2.
h) In this way, the biological reaction is advanced while stirring the biological culture solution 3 in the culture tank 2 with the culture tank agitator 11.
i) When the biological reaction has sufficiently progressed, the valve 13 is closed, the valve 12 and the valve 14 are opened, the culture tank pump 8 is driven, and the reaction product generated in the culture tank 2 is collected together with the filtrate A. Store in the filtrate storage tank 5.

The filter 4 includes a filtration membrane and a container that accommodates the filtration membrane. The filtration membrane may be an organic membrane or an inorganic membrane. The shape of the filtration membrane may be any shape such as a flat membrane, a hollow fiber membrane, and a spiral type. Among these, a hollow fiber membrane module is preferable. Any of the pressure type shapes can be employed.

As the filtration method, cross flow filtration using a hollow fiber membrane module is preferable. In this filtration method, a culture solution containing reaction products, microorganisms, and the like is filtered while being supplied to the inside of the hollow fiber membrane, and the filtrate is taken out from the outside. Microorganisms deposited inside the hollow fiber membrane And so on, so that a stable filtration state can be maintained over a long period of time.

When performing cross-flow filtration using a hollow fiber membrane module, it is necessary to flow the liquid to be filtered through the hollow fiber membrane at a flow rate of a certain level or more in order to scrape the membrane dirt. However, in the present invention, since the biological culture solution containing microorganisms and the like to be filtered contains oxygen-enriched MNB, even if it is flowed at a lower flow rate than usual, it is possible to scrape membrane dirt, Can significantly reduce the stress and damage to them.

Specifically, in general cross flow filtration, the soot circulation flow rate is about 1 to 2 m / s when an organic membrane is used, and about 1 to 3 m / s when a ceramic membrane is used. However, by adding oxygen-enriched MNB to the biological culture solution, it is possible to maintain low filtration resistance by reducing membrane contamination, so that the same flux (the amount of membrane filtered water per unit time / unit membrane area) can be obtained. The necessary circulation flow rate can be reduced to about 0.2 to 1.5 m / s. Further, when operating at the same circulation flow rate, the flux can be increased by about 1.2 to 2.0 times.

As the filtration membrane, an organic polymer compound can be suitably used from the viewpoints of separation performance, water permeability, and dirt resistance. Examples include polyethylene resins, polypropylene resins, polyvinyl chloride resins, polyvinylidene fluoride resins, polysulfone resins, polyethersulfone resins, polyacrylonitrile resins, cellulose resins, and cellulose triacetate resins. A mixture of these resins as the main component may be used. Polyvinyl chloride resins, polyvinylidene fluoride resins, polysulfone resins, polyethersulfone resins and polyacrylonitrile resins, which are easy to form in solution and have excellent physical durability and chemical resistance, are preferred. A vinylidene chloride resin or a resin containing the vinylidene fluoride resin as a main component is more preferably used because it has a characteristic of having both chemical strength (particularly chemical resistance) and physical strength.

Here, as the polyvinylidene fluoride-based resin, a homopolymer of vinylidene fluoride is preferably used. Furthermore, the polyvinylidene fluoride resin may be a copolymer of a vinyl monomer copolymerizable with vinylidene fluoride. Examples of vinyl monomers copolymerizable with vinylidene fluoride include tetrafluoroethylene, hexafluoropropylene, and ethylene trichloride fluoride.

The average pore diameter of the filtration membrane can be appropriately determined according to the purpose and situation of use, but it is preferably smaller to some extent, and is usually preferably 0.01 μm or more and 1 μm or less. When the average pore diameter of the hollow fiber membrane is less than 0.01 μm, components such as microorganisms, such as sugars and proteins, and membrane dirt components such as aggregates thereof block the pores, and stable operation cannot be performed. In consideration of the balance with water permeability, it is preferably 0.02 μm or more, and more preferably 0.03 μm or more. In addition, when it exceeds 1 μm, the film surface smoothness and the shearing force due to the flow of the film surface, and the peeling of dirt components from the pores by physical cleaning such as backwashing and air scrubbing are insufficient, and stable operation is possible. Disappear.

In addition, when the average pore diameter approaches the size of a microorganism or the like, these may directly block the pores. In addition, there may be cases where broken cells of microorganisms or cultured cells in the fermented liquid are killed to produce cell crushed materials. In order to avoid pore clogging by these crushed materials, the average pore diameter is 0.4 μm. The following is preferable, and 0.2 μm or less is preferable.

Here, the average pore diameter of the filtration membrane can be obtained by measuring and averaging the diameters of a plurality of pores observed by scanning electron microscope observation at a magnification of 10,000 times or more. Preferably, 10 or more, preferably 20 or more pores are randomly selected, the diameters of these pores are measured, and the number average is obtained. When the pores are not circular, it is also preferable to use an image processing device or the like to obtain a circle having an area equal to the area of the pores, that is, an equivalent circle, and obtain the equivalent circle diameter as the pore diameter. it can.

As shown in FIG. 1, in 1st Embodiment, the filtrate A which is the liquid made to contain MNB in the MNB generator 7a is driven out of the MNB generation tank 6 by driving the liquid supply pump 10, and the MNB generator In addition to being supplied to 7a, air C having an increased air concentration obtained by the oxygen enrichment means is supplied to the MNB generator 7a.

As the MNB generating apparatus 7a used in the first embodiment, as shown in FIG. 2, an apparatus that can generate a large amount of MNB economically and that is driven using a water flow (nozzle system) is used. In the MNB generator 7a, the filtrate A is supplied from the inlet 21 of the nozzle in a state where pressure is applied, and turbulence is generated in the throat 22 while reducing the diameter of the pipe and increasing the flow velocity. In this state, air C with increased oxygen concentration is supplied from the gas inlet 24, mixed with the filtrate A in the suction portion 23, becomes MNB by the water flow, and contains MNB of air with increased oxygen concentration from the outlet portion 25. The filtrate D to be discharged is discharged and supplied to the macro / nano bubble generating tank 6.
The amount and size of the MNB can be adjusted by adjusting the flow rate of the filtrate A and the air C with increased oxygen concentration supplied to the MNB generator 7a.

In the first embodiment, in order to obtain air C having an increased oxygen concentration supplied to the MNB generator 7a, oxygen enrichment means using an oxygen enriched film as shown in FIG. 3 is used. .
In this oxygen-enriching means using the oxygen-enriched film, basically, the container 31 in which the oxygen-enriched film 30 is disposed has an air introduction part 33 and a lead-out part for discharging air F having a low oxygen concentration at both ends. 34, the air pressurized by the intake fan 32 is vented from the air inlet 33 to the oxygen-enriched film 30, and the air C having an increased oxygen concentration is discharged from the outlet 35, and the oxygen concentration The low air F is discharged from the outlet 34.

In the first embodiment, the oxygen concentration of the gas constituting the MNB is made higher than the concentration in the air (about 21%), thereby reducing the amount of the biological culture solution extracted from the culture tank, and the biological culture solution contains Even if the amount of MNB to be reduced is decreased, the activity of the microorganisms and the like can be maintained by supplying the microorganisms with a high concentration of oxygen that is easily absorbed in the MNB state. Furthermore, by reducing the amount of the biological culture solution extracted from the culture tank, it is possible to reduce stress and damage to microorganisms and the like, and to reduce the energy required for circulation of the biological culture solution.

In the first embodiment, as the culture tank pump 8 and the return pump 9, positive displacement pumps such as a diaphragm pump, a tube pump, a screw pump, and a rotary pump that have relatively little stress and damage to microorganisms and the like are preferably used. This also makes it possible to further reduce the stress and damage to microorganisms.
Furthermore, by reducing the amount of MNB contained in the culture solution, the energy required for driving the MNB generator can be reduced.

○ Second embodiment (Fig. 4)
Next, a second embodiment of the present invention will be described with reference to FIG.
The second embodiment is a biological reaction apparatus for causing a microorganism or the like to generate a reaction product, and contains the oxygen-enriched MNB in the biological culture as follows.
a) The culture solution 1 is supplied to the culture tank 2.
b) The valve 15 is closed, the valve 16 is opened, and the culture tank pump 8 is driven to extract the biological culture solution 3 containing microorganisms and the like from the culture tank 2 and supply it to the micro / nano bubble generation tank 6.
c) The biological culture solution 3 is stored in the micro / nano bubble generating tank 6 and oxygen enriched MNB is contained by the micro / nano bubble generating device 7a.
d) The return pump 9 is driven to return the biological culture solution G containing the oxygen-enriched MNB to the culture tank 2.
e) In this way, the biological reaction is advanced while stirring the biological culture solution 3 in the culture tank 2 with the culture tank agitator 11.
f) When the biological reaction has sufficiently progressed, the valve 16 is closed, the valve 15 is opened and the culture tank pump 8 is driven, and the reaction product generated in the culture tank 2 is collected together with the filtrate A to obtain the filtrate. Store in storage tank 5.

The method of 1) (first embodiment) and the method of 2) (second embodiment), which are methods for adding oxygen-enriched MNB to the biological culture solution extracted from the culture tank, are the types of microorganisms and the like. It is preferable to adopt a method in which the stress and damage to the microorganisms are generally reduced depending on the conditions of the biological reaction.

In the present invention, as a means for containing an oxygen-enriched MNB in a biological culture solution, the biological culture solution extracted from the culture tank mentioned as the first feature point contains the oxygen-enriched MNB and is returned to the culture tank. Although means (hereinafter referred to as “first means”) is used, other means may be used in combination.

When the first means is used alone, it may take time to set the MNB content in the biological culture solution in the culture tank to an appropriate value. Includes means for containing oxygen-enriched MNB in the culture medium supplied to the culture tank (hereinafter referred to as “second means”), means for containing oxygen-enriched MNB in the biological culture medium in the culture tank (hereinafter referred to as “second means”). , "Third means") and the like. In particular, the second means is preferable as a means used in combination with the first means because the microorganisms and the like are not subjected to stress or damage due to the blowing of MNB.

○ Third embodiment (FIG. 5)
Next, a third embodiment of the present invention will be described with reference to FIG.
The third embodiment is a biological reaction apparatus for causing a microorganism or the like to generate a reaction product, and uses the second means in combination with the first embodiment (using the first means) of the present invention.
In the third embodiment, the oxygen-enriched MNB is contained in a culture solution such as a microorganism as follows.
a) The culture solution 1 supplied to the culture tank 2 is caused to contain oxygen-enriched MNB (culture solution E containing oxygen-enriched MNB) by the micro / nano bubble generator 7b.
b) In this way, the biological reaction is advanced while stirring the biological culture solution 3 in the culture tank 2 with the culture tank agitator 11.
c) When the content of micro-nano bubbles in the biological culture solution 3 at the beginning of the biological reaction is small, or when the content of micro-nano bubbles in the biological culture solution 3 decreases due to progress of the biological reaction, b in the first embodiment ) To g), by adding the oxygen-enriched MNB to the filtrate A obtained by filtering the biological culture solution 3 and returning to the culture tank 2, the content of micro-nano bubbles in the biological culture solution 3 Adjust to an appropriate value.
d) When the biological reaction is sufficiently advanced, the valve 13 is closed, the valve 12 and the valve 14 are opened, the culture tank pump 8 is driven, and the reaction product generated in the culture tank 2 is collected together with the filtrate A. Store in the filtrate storage tank 5.

○ Fourth embodiment (Fig. 6)
Next, a fourth embodiment of the present invention will be described with reference to FIG.
The fourth embodiment is a biological reaction apparatus for causing a microorganism or the like to generate a reaction product, and the second means and the third means are used in combination with the first embodiment (using the first means) of the present invention. Is.

In the fourth embodiment, the oxygen-enriched MNB is contained in a culture solution such as a microorganism as follows.
a) The culture solution 1 supplied to the culture tank 2 is caused to contain oxygen-enriched MNB (culture solution E containing oxygen-enriched MNB) by the micro / nano bubble generator 7b.
b) In this way, the biological reaction is advanced while stirring the biological culture solution 3 in the culture tank 2 with the culture tank agitator 11.
c) When the content of micro-nano bubbles in the biological culture solution 3 at the beginning of the biological reaction is small, or when the content of micro-nano bubbles in the biological culture solution 3 decreases due to progress of the biological reaction, b in the first embodiment ) To g), the filtrate A obtained by filtering the biological culture solution 3 is allowed to contain oxygen-enriched MNB and refluxed to the culture vessel 2, or the micronanobubble generator 7c is used to culture the culture vessel. By adding oxygen-enriched MNB to the biological culture solution 3 in 2, the content of the micro / nano bubbles in the biological culture solution 3 is adjusted to an appropriate value.
d) When the biological reaction is sufficiently advanced, the valve 13 is closed, the valve 12 and the valve 14 are opened, the culture tank pump 8 is driven, and the reaction product generated in the culture tank 2 is collected together with the filtrate A. Store in the filtrate storage tank 5.

As the third embodiment and the fourth embodiment of the present invention, the first embodiment (using the first means) of the present invention, which is the combination of the second means, the second means, and the third means, respectively, has been described. However, similarly, the second embodiment (using the first means) of the present invention can be used in combination with the second means, the second means, and the third means, respectively, and the same effects can be obtained. It can be easily understood by a contractor.

As described above, in the biological reaction apparatus of the present invention and the biological reaction method using this biological reaction apparatus, the oxygen concentration of the gas constituting the MNB is made higher than the concentration in air (about 21%). By reducing the amount of the biological culture solution withdrawn from the culture tank and reducing the amount of MNB contained in the biological culture solution, a high concentration of easily absorbed oxygen in the MNB state is supplied to the microorganism. The activity can be maintained.

Furthermore, by reducing the amount of the biological culture solution withdrawn from the culture tank, it is possible to reduce stress and damage to the microorganisms and to reduce the energy required for circulation of the biological culture solution.

In addition, as a pump for circulating a biological culture solution such as a pump for extracting the biological culture solution from the culture vessel, a pump for refluxing the biological culture solution containing oxygen-enriched MNB to the culture vessel, It is possible to use positive displacement pumps such as diaphragm pumps, tube pumps, screw pumps, rotary pumps, etc. that cause relatively little stress and damage to the This can be further reduced.

Furthermore, by reducing the amount of MNB contained in the culture solution, the energy required for driving the MNB generator can be reduced.
As described above, the present invention is excellent in that a biological reaction using microorganisms or the like can be performed efficiently and economically.

DESCRIPTION OF SYMBOLS 1 Culture liquid 2 Culture tank 3 Biological culture liquid containing culture liquid, microorganisms, etc. 4 Filter 5 Filtrate storage tank 6 Micro nano bubble generation tank 7a-7c Micro nano bubble generation apparatus 8 Culture tank pump 9 Return pump 10 Liquid supply pump 11 Culture Tank agitator 12 to 16 Valve 21 Inlet part 22 Throat part 23 Suction part 24 Gas inlet 25 Outlet part 30 Oxygen-rich film 31 Container 32 Intake fan 33 Air introduction part 34 (Exhaust air with low oxygen concentration) Deriving part 35 Derived part (discharges air with increased oxygen concentration) A Filtrate B Biological culture fluid excluding filtrate C Air with increased oxygen concentration D Filtrate containing oxygen-enriched MNB (filtrate + oxygen concentration Enhanced Air MNB)
E Culture medium containing oxygen-enriched MNB (culture medium + air MNB with increased oxygen concentration)
F Air with low oxygen concentration G Biological culture fluid containing oxygen-enriched MNB (biological culture fluid + MNB with increased oxygen concentration)
107 Culture tank as biological reaction tank 110 Cell filter 115 Micro-nano bubble generation tank 116 Micro-nano bubble generation device

Claims (12)

A culture vessel containing a culture solution and a biological culture solution containing microorganisms or cells;
A micro / nano bubble generating device containing micro / nano bubbles formed from air with an increased oxygen concentration in the biological culture solution extracted from the culture tank;
A conduit for refluxing the biological culture solution containing the micro / nano bubbles to the culture tank;
A biological reaction apparatus comprising: Between the culture tank and the micro-nano bubble generating device, a biological culture liquid extracted from the culture tank is disposed, and a filter for separating the filtrate and the biological culture liquid excluding the filtrate is disposed,
The filtrate contains micro-nano bubbles formed from air with an increased oxygen concentration by the micro-nano bubble generator,
The biological reaction apparatus according to claim 1, further comprising a conduit for refluxing the biological culture solution excluding the filtrate and the filtrate containing the micro / nano bubbles to the culture tank. The micro-nano bubbles are directly contained in a biological culture solution extracted from the culture tank without disposing a filter between the culture tank and the micro-nano bubble generator. Bioreactor. The biological reaction device according to any one of claims 1 to 3, wherein the micro / nano bubble generating device is of a system driven by using a water flow. A positive displacement pump such as a diaphragm pump, a screw pump, or a rotary pump is used as a pump for extracting the biological culture liquid from the culture tank and / or a pump for returning the biological culture liquid containing the micro-nano bubbles to the culture tank. The biological reaction apparatus according to any one of claims 1 to 4, which is used. The biological reaction apparatus according to claim 5, wherein the positive displacement pump is a tube pump. The biological reaction apparatus according to any one of claims 1 to 6, wherein the oxygen content of the air with an increased oxygen concentration is 25 to 40%. The biological reaction apparatus according to any one of claims 1 to 7, wherein the air having an increased oxygen concentration is obtained by passing air through an oxygen-enriched membrane. The air with an increased oxygen concentration is obtained by mixing oxygen generated by any of the PSA method, VSA method, cryogenic separation method, and chemical adsorption method with air using a line mixer or the like. The biological reaction apparatus according to any one of claims 1 to 7. The biological reaction apparatus according to any one of claims 1 to 9, further comprising a micro / nano bubble generating apparatus that contains micro / nano bubbles formed from air with an increased oxygen concentration in the culture solution supplied to the culture tank. The bioreactor according to any one of claims 1 to 10, further comprising a micro / nano bubble generating device that contains micro / nano bubbles formed from air with an increased oxygen concentration in the biological culture solution in the culture tank. A biological reaction method, characterized in that a reaction product such as a microorganism or cell metabolite is obtained or the microorganism or cell is propagated by the biological reaction device according to any one of claims 1 to 11.

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