WO2017029733A1 - Biological reaction device, and biological reaction method using said biological reaction device - Google Patents

Abstract

In this biological reaction device and this biological reaction method which uses said biological reaction device, micro-nanobubbles (MNB) are included in a biological culture solution extracted from a culture tank, a filter is used to separate the biological culture solution including the MNB into a filtrate and a concentrate of microorganisms or the like, the filtrate is recovered, and the concentrate of microorganisms or the like is returned to the culture tank. Accordingly, the amount of MNB included in the biological culture solution in the culture tank can be kept high, while reducing stress and damage incurred by the microorganisms or the like, and filtration and circulation of the biological culture solution are appropriately performed. As a result, a biological reaction using the microorganisms or the like can be performed efficiently and economically.

Description

Biological reaction apparatus and biological reaction method using the biological reaction apparatus

The present invention relates to a biological reaction apparatus for producing a reaction product (fermentation / brewing) by microorganisms or cells (hereinafter referred to as “microorganisms”), or for growing or concentrating (culturing) microorganisms, etc., and the biological reaction apparatus. In the case of micro-nano bubbles (hereinafter referred to as “MNB”, “micro-bubbles” as “MB”, and “nano-bubbles” as “NB”) The biological culture solution is filtered, and the filtrate is recovered and the biological culture solution (hereinafter referred to as “microorganism concentrate”) removed from the filtrate is returned to the culture tank. It is a feature.

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, in most cases, a reaction within one hour is sufficient for a chemical reaction, whereas in the case of a biological reaction, a reaction time of several days to several days or particularly several weeks or longer is required. In some cases. For this reason, it is required to perform biological reactions efficiently and economically.

Usually, (1) batch method (Batch method) and fed-batch method (Fed-Batch method) and (2) continuous method are used as methods for performing biological reactions (fermentation / brewing, culture) with microorganisms, etc. Specifically, a continuous method as disclosed in Patent Document 1 that can stably maintain a high yield and high productivity over a long period of time is employed.

In Patent Document 1, as shown in FIG. 5, in a continuous fermentation method, a fermentation broth in an apparatus is circulated between a fermentation reaction tank 101 and a membrane separation tank 112 by a fermentation broth circulation pump 111 to separate the separation membrane. The element 102 filters microorganisms and cultured cells through a separation membrane, collects the product from the filtrate, and at the same time refluxs the filtered microorganisms and cultured cells to the fermentation broth, increasing the concentration of the microorganisms and cultured cells in the fermentation broth. It is described that a high material productivity is obtained by maintaining.

Further, in Patent Document 2, in order to improve the biological reaction in culturing microorganisms and the like, the activation of microorganisms and the like is promoted by adding MNB or NB formed from air to the culture solution. It is disclosed that the reaction efficiency of the reaction, the reaction time, and the like are reduced.

Specifically, as shown in FIG. 6, the culture solution is extracted from the culture tank 207, filtered through the bacterial cell filter 210 to obtain a filtrate, and this filtrate is added to the MNB generation tank 215 by the MNB generator 216. A method for generating and mixing air MNB and returning it to the culture tank 207 is described.

However, in the method described in Patent Document 1 in which the biological culture solution is extracted from the culture tank and the concentrated liquid such as microorganisms is returned to the culture tank, the concentration of the microorganisms in the biological culture solution in the culture tank is maintained at a high level. However, on the other hand, since a biological culture solution having a high concentration such as microorganisms is filtered, the filtration membrane is likely to be clogged, and it is necessary to frequently wash or replace the filtration membrane. In addition, since a biological culture solution having a high concentration of microorganisms or the like is circulated outside the culture tank, oxygen necessary for respiration of microorganisms and the like is not sufficiently supplied in this route, and the microorganisms and the like are subjected to stress and damage.

Further, in the method described in Patent Document 2 in which a biological culture solution is extracted from a culture tank and air MNB is contained in the filtrate, the biological culture solution is filtered as compared with the amount of the biological culture solution extracted from the culture tank. The amount of filtrate obtained in this way is considerably small (the amount of filtrate is usually about 1/10 to 1/100 of the amount of biological culture liquid extracted from the culture tank). It is difficult to keep the amount of MNB contained in the biological culture solution in the culture tank high. Further, in order to increase the amount of the filtrate, increasing the amount of the biological culture solution extracted from the culture tank or increasing the filtration pressure will increase the stress and damage received by the microorganisms.

Furthermore, although the technical field is different from biological reactions such as fermentation, brewing, and culture, Patent Document 3 describes that in order to prevent clogging of the membrane module in the water purification system, It is disclosed that ultrafine bubbles are generated and water containing the ultrafine bubbles is supplied to the membrane module.

Specifically, as shown in FIG. 7, the raw water supplied from the raw water supply line 304 and / or the concentrated circulating water supplied from the membrane module 311 supplied from the concentrated circulating water line 317 are pressurized by the water supply pump 306. After that, a water purification method is described in which ultrafine bubbles are generated in water, and water containing the ultrafine bubbles is supplied to the membrane module 311 to perform membrane filtration.

However, in the water purification method as described in Patent Document 3 above, turbid substances, bacteria, and the like are simply removed from raw water such as river water by a filtration membrane such as a membrane module and filtered. Unlike the biological reaction in which microorganisms are returned to the culture tank and reused, there is no need to consider the stress and damage that the microorganisms receive.

Furthermore, the “ultrafine bubbles” used in Patent Document 3 are those having a bubble diameter of about 2 to 50 μm, which corresponds to MB and does not include NB, which is an ultrafine bubble having a diameter of 100 nm or less. is there.

Japanese Patent No. 5092487 Japanese Patent No. 4146476 Japanese Patent Application Laid-Open No. 2011-83764

The problem of the biological reaction apparatus of the present invention and the biological reaction method using this biological reaction apparatus is to maintain a high amount of MNB contained in the biological culture solution in the culture tank, to prevent clogging of the filtration membrane, The object is to reduce stress and damage received by microorganisms and the like, thereby efficiently and economically separating biological reactions and microorganisms using microorganisms.

In order to solve the above problems, in the biological reaction apparatus of the present invention and the biological reaction method using the biological reaction apparatus, MNB is contained in the biological culture solution extracted from the culture tank, and the biological culture solution containing the MNB is filtered. The filtrate is separated into a filtrate and a concentrate such as a microorganism by a vessel, and the filtrate is collected and the concentrate such as a microorganism is refluxed to a culture tank.

In addition, a means other than the above is used in combination with MNB in the biological culture solution, MNB formed from air with an increased oxygen concentration is used as MNB, and a volume as a pump for transporting a liquid containing microorganisms, etc. The above problem can be further solved by using a pH-type pump and adding a pH adjuster to the filtrate refluxed to the culture tank.

In the present invention, the amount of MNB contained in the biological culture solution in the culture tank can be maintained high, clogging of the filtration membrane can be prevented, and further, stress and damage to the microorganisms can be reduced. The biological reaction and microorganisms used can be separated efficiently and economically.

It is a schematic diagram which shows 1st Embodiment of the biological reaction apparatus of this invention. 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 1 (patent 5092487 gazette) which is a prior art example. It is FIG. 1 of patent document 2 (patent 4146476 gazette) which is a prior art example. It is FIG. 1 of patent document 3 (Unexamined-Japanese-Patent No. 2011-83764) 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.

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

The biological reaction apparatus of the present invention can be used for both biological reactions (fermentation and brewing) aimed at producing reaction products by microorganisms, and biological reactions aimed at the growth or concentration (culture) of microorganisms and the like. it can. The biological reaction apparatus of the present invention can be used for any of batch method (Batch method), fed-batch method (Fed-Batch method), and continuous method. It can be suitably used in a continuous process that can maintain the properties.

In biological reactions aimed at producing reaction products by microorganisms, the reaction products are continuously collected together with the filtrate, and in biological reactions aimed at the growth or concentration of microorganisms, microorganisms etc. Proliferate and concentrate to collect microorganisms.

The biological reaction of the present invention includes not only production of reaction products by microorganisms such as production of food, medicine, chemicals, etc. by brewing, fermentation, etc., production of bioethanol using biomass, but also growth or concentration of microorganisms, etc. It is also useful to apply to.

In the biological reaction apparatus of the present invention and the biological reaction method using this biological reaction apparatus, the biological culture liquid extracted from the culture tank is separated into a filtrate and a concentrated liquid such as microorganisms by a filter. Cross flow filtration using a hollow fiber membrane module can be suitably used.

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.

If a micro-nano bubble generator that uses water flow (nozzle method) is used, a large amount of MNB can be generated economically, stress and damage to microorganisms can be reduced, and clogging can be prevented. preferable.

In the biological reaction of the present invention, a reaction product is produced by a microorganism or the like in a biological culture solution containing the microorganism or the like contained in a culture tank, or the microorganism or the like is grown or concentrated.
As a nutrient source in the biological culture solution, one 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.

Although the concentration of microorganisms or cells added to the culture solution is not particularly limited, it is preferably 0.5 to 10.0 g / L, 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 for the first feature point of the present invention, the first feature point of the present invention is that, as described above, the biological culture solution extracted from the culture tank contains MNB, and the biological culture solution containing this MNB is filtered. Is separated into a filtrate and a concentrate such as microorganisms, and the filtrate is collected and the concentrate such as microorganisms is refluxed to the culture tank.
In the biological reaction in which the reaction product is produced (fermentation / brewing) by microorganisms or the like, or the microorganisms are propagated or concentrated (cultured),
(1) MNB is contained in a biological culture solution extracted from a culture tank containing a biological culture solution containing a culture medium or reaction raw materials, microorganisms, etc.
(2) The biological culture liquid containing MNB is separated into a filtrate and a concentrated liquid such as a microorganism by a filter,
(3) By collecting the filtrate and refluxing the concentrated liquid such as microorganisms to the culture tank, it is surprising that the amount of MNB contained in the biological culture liquid in the culture tank can be maintained high, and the filtration membrane. It is possible to prevent clogging of microorganisms, and furthermore, it is possible to reduce stress and damage received by microorganisms and the like, and thereby it is possible to efficiently and economically perform biological reactions using microorganisms, etc. It is.

As described above, in the continuous fermentation method in which a biological culture solution is extracted from a culture vessel and a concentrated solution such as microorganisms is returned to the culture vessel as described in Patent Document 1, a biological culture solution having a high concentration of microorganisms or the like is used. Is circulated outside the culture tank and filtered, and the filter membrane is likely to be clogged, and there is a problem that oxygen is not sufficiently supplied to the microorganisms in the circulation path. The biological culture solution contains MNB, the biological culture solution containing MNB is supplied to a filter, and separated into a filtrate and a concentrated liquid such as a microorganism. Such a problem can be solved by returning to the culture tank.

That is, in the present invention, by filtering the biological culture solution containing MNB with a filter, the MNB is interposed between the filter membrane and the substance (microorganism, turbid substance, etc.) contained in the biological culture solution. In addition, since it acts to prevent the substance from adhering to the filtration membrane, clogging of the filtration membrane can be prevented.

Thus, for example, when performing cross-flow filtration using a hollow fiber membrane, in order to perform filtration while stripping off substances adhering to the hollow fiber membrane surface from the membrane surface, the filtration flow rate is usually set against the membrane cross-sectional area. However, when filtering a biological culture solution containing MNB as in the present invention, it adheres to the hollow fiber membrane surface even if the filtration flow rate is 0.5 m / s or less. Since the filtration can be carried out while sufficiently stripping off the substance to be removed, the stress and damage to the microorganisms and the like in the filtration process can be reduced.

Furthermore, since this improves filtration efficiency, the size of the filtration device can be reduced, and the amount of biological culture solution supplied to the filtration device can be reduced, thereby reducing the equipment and operating costs of the biological reaction device. In addition, it is possible to reduce stress and damage to microorganisms.

Furthermore, in the present invention, by adding MNB to the biological culture solution extracted from the culture tank, oxygen can be sufficiently supplied to the microorganisms and the like contained in the biological culture solution circulated through the circulation path. The stress and damage received can be reduced.

Further, as described above, in the method of adding MNB to the filtrate of the biological culture solution extracted from the culture tank as described in Patent Document 2, the amount of the biological culture solution extracted from the culture tank is smaller than this. Since the amount of the filtrate obtained by filtering the biological culture solution is considerably small (the amount of the filtrate is usually about 1/10 to 1/100 of the amount of the biological culture solution extracted from the culture vessel), There is a problem that it is difficult to maintain a high amount of MNB contained in the biological culture solution, and in order to solve this problem, if the amount of the biological culture solution extracted from the culture tank is increased or the filtration pressure is increased. However, in the present invention, MNB is contained in the biological culture liquid extracted from the culture tank, and the biological culture liquid containing MNB is filtered. Supply to, and separated into a filtrate and a microorganism concentrates, as well as collecting the filtrate, in order to reflux the microorganisms concentrate the culture tank, it is possible to solve such a problem.

That is, in the present invention, the biological culture liquid extracted from the culture tank contains MNB, and the biological culture liquid containing this MNB is supplied to the filter without causing extra stress or damage to microorganisms. The MNB can be sufficiently contained in the biological culture medium circulated through the circulation path, and the amount of MNB contained in the biological culture liquid in the culture tank can be maintained high.

About the 2nd characteristic point of this invention The 2nd characteristic point of this invention is MNB to the biological culture liquid extracted from the culture tank mentioned as a 1st characteristic point as a means to make a biological culture liquid contain MNB. The biological culture solution containing MNB is separated into a filtrate and a concentrate such as a microorganism by a filter, and the filtrate is recovered and the microorganisms concentrate is returned to the culture tank (hereinafter referred to as “No. Together with other MNB-containing means.

When the first means is used alone, when it takes time to set the MNB content of the biological culture in the culture tank to an appropriate value, it is necessary to shorten this time. Include means for containing MNB in the culture medium supplied to the culture tank (hereinafter referred to as “second means”), means for containing MNB in the biological culture solution in the culture tank (hereinafter referred to as “third means”). It is preferable to use a means such as 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.

About the 3rd characteristic point of this invention The 3rd characteristic point of this invention is using MNB which raised oxygen concentration as MNB contained in a biological culture solution.
Thereby, 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 is reduced, the high concentration of oxygen that is easily absorbed in the MNB state is reduced to microorganisms in the culture tank. Can maintain the activity of microorganisms and 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 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, from an economical point of view, it is preferable to obtain air with an increased oxygen concentration by using an oxygen-enriched membrane and passing air through the oxygen-enriched membrane. Moreover, when using PSA method, VSA method, and chemical adsorption method, it is preferable to obtain air with an increased oxygen concentration by mixing oxygen produced by these methods and air with a line mixer or the like.

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.

About the 4th characteristic point of this invention The 4th characteristic point of this invention is a diaphragm pump as a pump which conveys the biological culture liquid containing microorganisms etc., such as a pump for extracting the biological culture liquid from the said culture tank. , Tube pumps, screw pumps, rotary pumps and other positive displacement pumps.
By using such a positive displacement pump to transport a biological culture solution containing microorganisms or the like, it is possible to further reduce stress and damage to the microorganisms and the like.

About the 5th feature point of this invention The 5th feature point of this invention is providing the means which adds a pH adjuster to the pipe line which circulates a filtrate to a culture tank.
Microorganisms and the like produce by-products such as organic acids in the biological reaction in the culture tank. If this changes the pH of the biological culture solution in the culture tank, this is within the range suitable for the microorganism or the like. It needs to be adjusted.

As a general pH adjusting means, a pH adjusting agent such as acid or alkali is directly added to a biological culture solution in a culture tank. However, this means locally increases the concentration of acid / alkali, and prevents microorganisms, etc. It will cause stress and damage.
In the present invention, since a pH adjuster such as acid or alkali is added to the filtrate refluxed to the culture tank, compared to the case where the pH adjuster is added directly to the biological culture liquid, The pH of the biological culture solution in the culture tank can be adjusted without causing stress or damage due to the concentration difference.

* 1st Embodiment (FIG. 1)
First, a first embodiment of the present invention will be described with reference to FIG.
The biological reaction apparatus of the first embodiment is used for both biological reactions (fermentation and brewing) aimed at producing reaction products by microorganisms, and biological reactions aimed at growing or concentrating (culturing) microorganisms. be able to. When the reaction product is intended to be produced, the reaction product is continuously collected together with the filtrate, and when the microorganism is intended to grow or concentrate, after the microorganism is grown, Microorganisms and the like are collected by concentrating the biological culture solution in the culture tank.

First, a case where a biological reaction for the purpose of generating a reaction product by a microorganism or the like is performed will be described.
Normally, the valve 10 is closed and the valve 11 is opened, and a biological reaction is performed by the following process.
a) While stirring with the culture tank stirrer 7, the biological reaction is performed in the biological culture solution 3 containing the culture medium or the reaction raw material 1, microorganisms, and the like stored in the culture tank 2.
b) While performing the reaction of a) above, the pump 8 is driven to continuously extract the biological culture solution 3 after the reaction from the culture tank 2 and supply it to the micro / nano bubble generator 6a to contain the MNB of air A Let
c) The biological culture liquid containing MNB of air A is supplied to the filter 4 and separated into the filtrate B containing the reaction product and the concentrated liquid C such as microorganisms.
e) The filtrate B is collected in the filtrate storage tank 5 and the concentrated liquid C such as microorganisms containing MNB is refluxed to the culture tank 2.
In addition, in order to keep the amount of the culture solution and microorganisms in the culture tank 2 constant, the opening and closing of the valve 10 and the valve 11 are adjusted to adjust the amount of the filtrate B refluxed to the culture tank 2, or newly Operations such as supplying the culture medium or reaction raw material 1 and microorganisms to the culture tank 2 are appropriately performed.

Next, a case where a biological reaction for the purpose of growing or concentrating microorganisms is performed will be described.
As a first stage, until a microorganism or the like grows to an appropriate amount, an operation is performed in the same process as a biological reaction aimed at generating a reaction product by the microorganism or the like. By separating and collecting the filtrate B, by-products such as organic acids generated by biological reactions such as microorganisms can be collected together with the filtrate B. In this first stage, the amount of the filtrate B refluxed to the culture tank 2 is adjusted by adjusting the opening and closing of the valve 10 and the valve 11 in the same manner as the biological reaction aimed at generating the reaction product by the microorganisms and the like. be able to.

As a second stage, after microorganisms and the like have grown to an appropriate amount, the supply of the culture medium or reaction raw material 1 to the culture tank 2 is stopped and the valve 10 is closed to recirculate the filtrate B to the culture tank 2. Stop and concentrate / recover microorganisms by the following process.
a) While growing microorganisms and the like, the pump 8 is driven to continuously extract the biological culture solution 3 after the reaction from the culture tank 2 and supply it to the micro / nano bubble generator 6a to contain the MNB of air A.
b) The biological culture solution containing MNB of air A is supplied to the filter 4 and separated into a filtrate B containing a reaction product and a concentrated liquid C such as a microorganism.
c) The filtrate B is collected in the filtrate storage tank 5, and the concentrated liquid C such as microorganisms containing MNB is refluxed to the culture tank 2.
As described above, in the second stage, the biological culture liquid having a high concentration of microorganisms or the like is circulated outside the culture tank 2 via the micro / nano bubble generating device 6a and the filter 4. Since MNB is contained in the biological culture solution 3 extracted from the culture tank 2, clogging of the filtration membrane can be prevented, oxygen can be sufficiently supplied to the microorganisms in the circulation path, and the stress received by the microorganisms as a result.・ Damage can be reduced.

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 into the hollow fiber membrane at a flow rate of a certain level or more in order to scrape membrane dirt. Since the biological culture solution containing microorganisms, etc., which is subject to filtration, contains MNB, even if it is flowed at a lower flow rate than usual, it can sufficiently scrape membrane dirt, and stress and damage to microorganisms etc. Can be greatly reduced.

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 including MNB in the biological culture solution, the membrane contamination is reduced and the filtration resistance can be kept small, so the circulation necessary to obtain the same flux (the amount of membrane filtration water per unit time and unit membrane area) The 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. If 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. Furthermore, a part of microorganisms or the like may be killed to produce a crushed material. In order to avoid pore clogging by these crushed materials, the average pore diameter is preferably 0.4 μm or less, and 0.2 μm. The following are preferred.

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.

○ Second embodiment (Fig. 2)
A second embodiment of the present invention will be described with reference to FIG.
The biological reaction apparatus of the second embodiment is a combination of the second embodiment and the first embodiment (using the first means) of the present invention. In the second embodiment, as in the first embodiment, biological reactions (fermentation / brewing) aimed at producing reaction products by microorganisms, biological reactions aimed at growing or concentrating (culturing) microorganisms, etc. Any of them can be used.

First, a case where a biological reaction for the purpose of generating a reaction product by a microorganism or the like is performed will be described.
Normally, the valve 10 is closed and the valve 11 is opened, and a biological reaction is performed by the following process.
a) The culture medium or reaction raw material 1 is supplied to the micro / nano bubble generator 6b to contain the MNB of air A, and the culture medium or reaction raw material D containing the MNB of air A is supplied to the culture tank 2.
b) The biological reaction is performed in the biological culture solution 3 containing the culture medium or the reaction raw material 1 and microorganisms stored in the culture tank 2 while being stirred by the culture tank agitator 7.
c) While performing the reaction of b) above, the pump 8 is driven to continuously extract the biological culture solution 3 after the reaction from the culture tank 2 and supply it to the micro / nano bubble generator 6a to contain the MNB of air A Let
d) The biological culture solution containing the MNB of air A is supplied to the filter 4 and separated into the filtrate B containing the reaction product and the concentrated liquid C such as microorganisms.
e) The filtrate B is collected in the filtrate storage tank 5 and the concentrated liquid C such as microorganisms containing MNB is refluxed to the culture tank 2.
In addition, in order to keep the amount of the culture solution and microorganisms in the culture tank 2 constant, the opening and closing of the valve 10 and the valve 11 are adjusted to adjust the amount of the filtrate B refluxed to the culture tank 2, or newly Operations such as supplying the culture medium or reaction raw material 1 and microorganisms to the culture tank 2 are appropriately performed.

Next, a case where a biological reaction for the purpose of growing or concentrating microorganisms is performed will be described.
As a first stage, until a microorganism or the like grows to an appropriate amount, an operation is performed in the same process as a biological reaction aimed at generating a reaction product by the microorganism or the like. By separating and collecting the filtrate B, by-products such as organic acids generated by biological reactions such as microorganisms can be collected together with the filtrate B. In this first stage, the amount of the filtrate B refluxed to the culture tank 2 is adjusted by adjusting the opening and closing of the valve 10 and the valve 11 in the same manner as the biological reaction aimed at generating the reaction product by the microorganisms and the like. be able to.

As a second stage, after microorganisms and the like have grown to an appropriate amount, the supply of the culture medium or reaction raw material 1 to the culture tank 2 is stopped and the valve 10 is closed to recirculate the filtrate B to the culture tank 2. Stop and concentrate / recover microorganisms by the following process.
a) While growing microorganisms and the like, the pump 8 is driven to continuously extract the biological culture solution 3 after the reaction from the culture tank 2 and supply it to the micro / nano bubble generator 6a to contain the MNB of air A.
b) The biological culture solution containing MNB of air A is supplied to the filter 4 and separated into a filtrate B containing a reaction product and a concentrated liquid C such as a microorganism.
c) The filtrate B is collected in the filtrate storage tank 5, and the concentrated liquid C such as microorganisms containing MNB is refluxed to the culture tank 2.
As described above, in the second stage, the biological culture liquid having a high concentration of microorganisms or the like is circulated outside the culture tank 2 via the micro / nano bubble generating device 6a and the filter 4. Since MNB is contained in the biological culture solution 3 extracted from the culture tank 2, clogging of the filtration membrane can be prevented, oxygen can be sufficiently supplied to the microorganisms in the circulation path, and the stress received by the microorganisms as a result.・ Damage can be reduced.

○ Third embodiment (FIG. 3)
A third embodiment of the present invention will be described with reference to FIG.
The biological reaction apparatus according to the third embodiment is a combination of the second means and the third means in the first embodiment (using the first means) of the present invention.
In the third embodiment, as in the first embodiment, biological reactions (fermentation / brewing) aimed at producing reaction products by microorganisms, and biological reactions aimed at growing or concentrating (culturing) microorganisms etc. Any of them can be used.

First, a case where a biological reaction for the purpose of generating a reaction product by a microorganism or the like is performed will be described.
Normally, the valve 10 is closed and the valve 11 is opened, and a biological reaction is performed by the following process.
a) The culture medium or reaction raw material 1 is supplied to the micro / nano bubble generator 6b to contain the MNB of air A, and the culture medium or reaction raw material D containing the MNB of air A is supplied to the culture tank 2.
b) The culture medium or reaction raw material 1 stored in the culture tank 2 while being fed to the culture tank 2 while the biological culture solution E stirred with the culture tank agitator 7 and containing the MNB of air A with the micro-nano bubble generator 6c is supplied. The biological reaction is performed in the biological culture solution 3 containing microorganisms and the like. The biological culture solution 3 is supplied to the micro / nano bubble generating device 6 c by extracting the biological culture solution 3 from the culture tank 2 by the pump 9.
c) While performing the reaction of b) above, the pump 8 is driven to continuously extract the biological culture solution 3 after the reaction from the culture tank 2 and supply it to the micro / nano bubble generator 6a to contain the MNB of air A Let
d) The biological culture solution containing the MNB of air A is supplied to the filter 4 and separated into the filtrate B containing the reaction product and the concentrated liquid C such as microorganisms.
e) The filtrate B is collected in the filtrate storage tank 5 and the concentrated liquid C such as microorganisms containing MNB is refluxed to the culture tank 2.
In addition, in order to keep the amount of the culture solution and microorganisms in the culture tank 2 constant, the opening and closing of the valve 10 and the valve 11 are adjusted to adjust the amount of the filtrate B refluxed to the culture tank 2, or newly Operations such as supplying the culture medium or reaction raw material 1 and microorganisms to the culture tank 2 are appropriately performed.

Next, a case where a biological reaction for the purpose of growing or concentrating microorganisms is performed will be described.
As a first stage, until a microorganism or the like grows to an appropriate amount, an operation is performed in the same process as a biological reaction aimed at generating a reaction product by the microorganism or the like. By separating and collecting the filtrate B, by-products such as organic acids generated by biological reactions such as microorganisms can be collected together with the filtrate B.

As a second stage, after the microorganisms and the like have grown to an appropriate amount, the valve 10 is closed to stop the reflux of the filtrate B to the culture tank 2, and the microorganisms and the like are concentrated and recovered by the following steps.
a) While performing the growth, the pump 8 is driven to continuously extract the biological culture solution 3 from the culture tank 2, and is supplied to the micro / nano bubble generating device 6a to contain the MNB of the air A. Driven and supplied to the filter 4.
b) In the filter 4, the biological culture solution is separated into the filtrate B and the concentrated liquid C such as microorganisms.
c) The filtrate B is collected in the filtrate storage tank 5, and the concentrated liquid C such as microorganisms containing MNB is refluxed to the culture tank 2.
As described above, in the second stage, a biological culture solution having a high concentration of microorganisms or the like is circulated outside the culture tank 2 through the filter 4. In the present invention, the biological culture extracted from the culture tank 2 is used. Since the liquid 3 contains MNB, clogging of the filtration membrane can be prevented, oxygen can be sufficiently supplied to the microorganisms and the like in the circulation path, and stress and damage to the microorganisms can be reduced.

○ Fourth embodiment (Fig. 4)
A fourth embodiment of the present invention will be described with reference to FIG.
The biological reaction apparatus according to the fourth embodiment is provided with means for adding the pH adjusting agent 12 to the filtrate B refluxed to the culture tank 2 in the first embodiment of the present invention. By adjusting the addition amount of the pH adjuster 12, the pH of the biological culture solution 3 in the culture tank 2 can be adjusted to a range suitable for microorganisms and the like.

In the fourth embodiment, as in the first embodiment, biological reactions (fermentation / brewing) for the purpose of producing reaction products by microorganisms, biological reactions for the purpose of growth or concentration (culture) of microorganisms, etc. Any of them can be used.

In the first embodiment, normally, the valve 10 is closed and the valve 11 is opened to perform a biological reaction. However, in order to keep the amount of the biological culture solution 3 and microorganisms in the culture tank 2 constant, the valve 10 Is opened, and part or all of the filtrate B separated by the filter 4 is refluxed to the culture tank 2.
At that time, in the fourth embodiment, the pH adjuster 12 is added by the following steps.
a) From the state where the valve 10 is closed and the valve 11 is opened and the biological reaction is being performed, the valve 10 is opened and a part or all of the filtrate B separated by the filter 4 is removed from the pH adjuster mixing tank 13. To supply.
b) The amount of pH adjuster 12 (acid, alkali) necessary for adjusting the pH of the biological culture solution 3 in the culture tank 2 to a range suitable for microorganisms or the like in the filtrate B in the pH adjuster mixing tank 13 Etc.) is added.
c) The filtrate B to which the pH adjuster 12 has been added is refluxed to the culture tank 2.

When a pH adjusting agent such as acid or alkali is directly added to the biological culture solution 3 in the culture tank 2, the acid / alkali concentration locally increases, and this causes stress and damage to microorganisms. However, by adding the pH adjusting agent as described above, the pH of the biological culture solution 3 in the culture tank 2 is adjusted without causing stress or damage associated with the concentration difference to the microorganisms or the like in the culture tank 2. be able to.

As described above, in the bioreaction apparatus of the present invention and the bioreaction method using this bioreaction apparatus, stress damage caused to microorganisms or the like is caused by filtering the biological culture solution containing MNB with a filter. The amount of MNB contained in the biological culture solution in the culture tank can be maintained high, and the biological culture solution can be filtered and circulated appropriately, thereby enabling efficient biological reaction using microorganisms and the like. It can be done efficiently and economically.

In the present invention, as means for containing MNB in the biological culture solution, the MNB of the biological culture solution in the culture tank can be obtained in a short time by using the first means together with the second means and the third means. The content of can be set to an appropriate value.

Moreover, in this invention, the amount of the biological culture liquid extracted from a culture tank is reduced by using MNB formed from the air which raised oxygen concentration as MNB contained in a biological culture liquid, and biological culture liquid contains Even if the amount of MNB is decreased, high concentration oxygen that is easily absorbed can be supplied to microorganisms in the culture tank, reducing the stress and damage to microorganisms and maintaining the activity of microorganisms. In addition, the energy required for the circulation of the biological culture solution and the energy required for driving the MNB generator can be reduced.

In the present invention, 1) a pump for extracting a biological culture solution from the culture tank, 2) a pump for supplying a biological culture solution containing micro-nano bubbles from a micro-nano bubble generator to a filter, and 3) culture. Diaphragm pumps, tube pumps, screw pumps, and rotary pumps that deliver relatively little stress and damage to microorganisms, such as pumps for transporting liquids containing microorganisms, such as pumps for refluxing biological culture solutions without filtrate. By using a positive displacement pump such as a pump, it is possible to further reduce the stress and damage to the microorganisms and maintain the activity of the microorganisms.

Further, in the present invention, a means for adding a pH adjuster is provided in a conduit for refluxing the filtrate to the culture tank, and by adding a pH adjuster such as acid or alkali to the filtrate refluxed to the culture tank. The pH of the biological culture solution in the culture tank can be adjusted without giving stress or damage due to the concentration difference to the microorganisms in the culture tank.

DESCRIPTION OF SYMBOLS 1 Medium or reaction raw material 2 Culture tank 3 Biological culture solution 4 Filter 5 Filtrate storage tank 6a-6c Micro nano bubble generator 7 Culture tank agitator 8, 9 Pump 10, 11 Valve 12 pH adjuster 13 pH adjuster mixing tank A Air B Filtrate C Concentrated liquid such as microorganism D Medium or reaction raw material containing MNB of air A E Biological culture solution containing MNB of air A 101 Fermentation reaction tank 102 Separation membrane element 111 Fermentation culture medium circulation pump 112 Membrane Separation tank 207 Culture tank 210 Cell filter 215 MNB generation tank 216 MNB generator 304 Raw water supply line 306 Water supply pump 311 Membrane module 317 Raw water and / or concentrated circulating water line

Claims (11)

A culture vessel containing a culture medium or reaction raw material and a biological culture solution containing microorganisms or cells;
A micro-nano bubble generator for containing micro-nano bubbles in a biological culture liquid extracted from the culture tank;
Having a filter that separates the biological culture solution containing the micro-nano bubbles into a biological culture solution excluding the filtrate and the filtrate;
A conduit for collecting the filtrate, a conduit for refluxing the filtrate to the culture tank, and a conduit for refluxing the biological culture liquid excluding the filtrate to the culture tank,
A biological reaction apparatus for producing reaction products by microorganisms or cells, or for growing or concentrating microorganisms or cells. The bioreaction apparatus according to claim 1, further comprising a micro / nano bubble generating apparatus for adding micro / nano bubbles to the culture solution supplied to the culture tank. The bioreaction apparatus according to claim 1 or 2, further comprising a micro / nano bubble generation apparatus that contains micro / nano bubbles in the biological culture solution in the culture tank. The biological reaction device according to any one of claims 1 to 3, wherein the micro-nano bubbles are micro-nano bubbles formed from air with an increased oxygen concentration. The biological reaction apparatus according to claim 4, wherein the oxygen content of the air with an increased oxygen concentration is 25 to 40%. The biological reaction apparatus according to claim 4 or 5, wherein the air having an increased oxygen concentration is obtained by passing air through an oxygen-enriched membrane. 6. The air according to claim 4 or 5, wherein the air having an increased oxygen concentration is obtained by mixing oxygen produced by any one of the PSA method, the VSA method, and the chemical adsorption method with air using a line mixer or the like. The biological reaction apparatus as described. A positive displacement pump such as a diaphragm pump, a screw pump, or a rotary pump is used as a pump for transporting a biological culture solution containing microorganisms or cells, such as a pump for extracting a biological culture solution from the culture tank. A biological reaction device according to any one of the above. The biological reaction apparatus according to claim 8, wherein the positive displacement pump is a tube pump. The biological reaction apparatus according to any one of claims 1 to 9, further comprising means for adding a pH adjusting agent to a conduit for refluxing the filtrate to the culture tank. A biological reaction method comprising producing a reaction product by microorganisms or cells, or growing or concentrating microorganisms or cells by the biological reaction apparatus according to any one of claims 1 to 10.

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