JP2922202B1 - Liquid solvent separation method and apparatus in microorganism culturing apparatus - Google Patents

Abstract

Abstract: PROBLEM TO BE SOLVED: To efficiently separate and collect a liquid solvent in a microorganism culture solution by using a porous filtration separation membrane. SOLUTION: A number of porous filtration / separation membranes 9 are arranged in a gas phase space 7 above a culture vessel 1, and a wave-making blade 10 is arranged below the culture vessel 1, and the blade 10 is reciprocated. As a result, the microbial culture solution 8 contained in the culture vessel 1 is swung to force the microbial culture solution 8 into contact with the porous filtration / separation membrane 9 in the gas phase space 7, and the clogged substance is washed by the impact of the forced contact to remove the eye. While preventing clogging, the liquid solvent is separated and recovered from the microorganism culture solution 8 by capillary action of the separation membrane 9.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a microorganism culturing apparatus, and more particularly to a method and apparatus for separating a liquid solvent in a microorganism culturing apparatus for separating a liquid solvent from a microorganism culture solution in a culture vessel.

[0002]

A method and an apparatus for separating a liquid solvent from a microorganism culture solution in a culture vessel by using a porous filtration separation membrane have been disclosed by one of the present inventors, for example, in Japanese Patent Laid-Open No. No. 95778, JP-A-6
−98758, etc.
A porous filtration membrane is arranged as a membrane, and the liquid solvent in the microbial culture is passed through, for example, by a pressure difference due to aeration, and the separation and recovery are performed. Solid fine particles and insoluble powders that cannot pass through become clogging substances, which cause clogging of the porous filtration separation membrane, thereby resulting in a decrease in filtration flow rate and failing to maintain good filtration efficiency. The possibility remains.

[0003] In order to prevent the clogging and a reduction in the filtration flow rate due to the clogging, for example, a backwashing device for performing backwashing intermittently or as necessary from the non-culture solution side is installed, or diatoms are added to the microorganism culture solution. It is possible to take measures such as adding a filter aid such as soil. However, in this case, the equipment cost for filtration is high, the running cost is high, and the operation and addition work are required. Although it is necessary to complicate the management, these methods also do not provide a drastic measure against clogging. Therefore, in this seed culture apparatus, filtration of the porous filtration separation membrane and clogging accompanying the filtration are performed. It is desired to eliminate the relationship and maintain good filtration efficiency.

[0004] The present invention has been made in view of the above circumstances, and an object of the present invention is to perform filtration of a porous filtration membrane as efficiently as possible and to eliminate clogging accompanying the filtration. Another object of the present invention is to provide a method and an apparatus for separating a liquid solvent in a microorganism culturing apparatus capable of maintaining filtration efficiency as good as possible.

[0005]

Means for Solving the Problems In view of the above problems, the present inventors have conducted intensive studies, and as a result, as described in JP-A-5-95778, for example, as a porous filtration separation membrane, Ceramic membrane, porous glass membrane, porous organic polymer membrane, woven metal fiber, sintered metal, etc. can be used, but these separation membranes act as a filter for liquid solvent separation. However, focusing on the fact that each of them exhibits a capillary phenomenon (capillary phenomenon) with respect to the liquid, a gas phase space is formed above the culture vessel containing the microorganism culture solution, and the gas phase space is formed. A porous filtration separation membrane is arranged, and the microbial culture solution is forcibly brought into contact with the porous filtration separation membrane in the gas-phase space. The medium can be absorbed and passed, and the microorganism culture solution can be filtered well. At this time, clogging substances such as the solid fine particles and insoluble powder in the microorganism culture solution that cannot pass through the separation membrane are removed. May adhere to the surface of the microbe, but the porous filtration membrane is washed while receiving a contact impact on the surface due to the forced contact of the microbial culture medium. At the same time, it falls back into the microbial culture solution and forms a natural washing cycle to eliminate the clogging without adhering to the surface of the separation membrane without causing clogging. Eliminate clogging during filtration,
It has been found that it is extremely effective in maintaining a high filtration efficiency.

[0006] The present invention has been made based on such knowledge, that is, the present invention according to claim 1 is directed to providing a method for separating a liquid solvent from the above-mentioned problems by using an upper gas phase in a culture vessel. The microbial culture solution contained in the culture container is forcibly brought into contact with the porous filtration separation membrane arranged in the space, and the liquid solvent in the microbial culture solution is separated and recovered by absorption passage of the capillary phenomenon in the porous filtration separation membrane. A method for separating a liquid solvent in a microorganism culturing apparatus, characterized in that, in addition to the above, in addition to the above, the filtration of the liquid solvent by the porous filtration separation membrane is promoted to ensure sufficient filtration performance. 2. The method according to claim 1, wherein the porous filtration separation membrane in the gas phase space is a hollow pipe-shaped separation membrane, and the hollow portion is continuously or intermittently pressurized.
According to a third aspect of the present invention, there is provided a method for separating a liquid solvent in a microbial culture apparatus, comprising the steps of: A porous filtration membrane arranged in a space,
A liquid solvent separation device in a microorganism culture device, comprising: forced contact means for forcibly bringing the microorganism culture solution contained in the culture container into contact with the porous filtration separation membrane in the gas phase space. In addition to the above, the inventions described in Claims 4 and 5 are intended to perform simple and reliable forced contact of the microorganism culture solution with the porous filtration / separation apparatus, thereby providing preferable forced contact means. The invention is characterized in that the forcible contacting means is constituted by wave-making means for wave-forming and oscillating a microorganism culture solution in a culture vessel, wherein the liquid solvent separation device in the microorganism culturing apparatus according to claim 1, The invention according to claim 5 is characterized in that the forcible contact means is constituted by a droplet forming means for ejecting or scattering a microorganism culture solution in a culture vessel into a gas phase space. The liquid solvent separating apparatus in the microorganism culturing apparatus according to the first aspect, wherein the invention according to the sixth aspect further enhances the filtration of the liquid solvent by the porous filtration separation membrane to secure sufficient filtration performance in addition to the above. To be able to
The porous filtration separation membrane is formed as a hollow pipe-shaped separation membrane, and a pressure reducing means for continuously or intermittently reducing the pressure of the hollow portion is further provided. A liquid solvent separating device in the microorganism culturing device described in 3, 4, or 5, which is used as a means for solving the above problem as a gist of the invention.

[0007]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described in more detail with reference to the drawings. In FIGS. 1 and 2, A denotes a microorganism culturing apparatus, B denotes a liquid solvent separating apparatus, and B denotes a liquid solvent separating apparatus. Is a porous filtration separation membrane 9 arranged in the culture vessel 1, a gas phase space 7 above the culture vessel 1, and a microbial culture solution 8 contained in the culture vessel 1. And a forced contact means for forcibly contacting the membrane 9. In the present embodiment, the forced contact means is provided by a wave forming means for wave-forming and moving the microorganism culture solution 8 in the culture vessel 1. There are things.

In this embodiment, the culture vessel 1 has, for example, detachably fixed closed side plates at both ends of a cylindrical body with bolts.
The lower semicircular portion of the cylindrical body is set to an appropriate volume of, for example, about 30 to 100 liters so as to accommodate the microbial culture solution 8 so that the liquid surface is at the middle position between the upper and lower sides, particularly the center position. The circular portion is a gas-phase space 7 above the culture solution accommodating portion, and is, for example, a stainless-steel horizontal cylinder.

In this embodiment, the porous filtration separation membrane 9 is a hollow pipe-shaped separation membrane, for example, a porous ceramic separation membrane having a pore diameter of 0.2 μm from the above various materials, that is, a ceramic porous separation membrane. In this example, the separation membranes are arranged in a gas phase space, and in the gas phase space 7, the separation membranes are arranged so as to be spaced above the liquid surface of the microorganism culture solution 8 so as to cover them. It is detachably mounted in the culture vessel 1 in the direction intersecting the cylinder so as to make the forcible contact of the microorganism culture solution 8 described later as efficiently as possible. In this example, a plurality of pieces are arranged in parallel in parallel in close proximity to a plurality of positions slightly spaced apart from each other in the longitudinal direction, and the pieces are arranged stepwise up and down. Lever arrangement
For example, the efficiency of the above-mentioned forced contact is ensured by using the lower three wires and the upper two wires each at two locations.

The porous filtration / separation membrane 9 is connected to a filtrate tank (not shown) via a tube or pipe 3 connected to one end in the longitudinal direction thereof so as to communicate with the hollow portion. Similarly, a pump (not shown) is interposed in the middle in the longitudinal direction of the filter 3 and the pump is used as a pressure reducing means, whereby the hollow portion of each of the porous filtration separation membranes 9 is continuously or intermittently compressed. In this embodiment, the pressure is reduced intermittently by using a tubing pump, for example.

In this embodiment, the wave forming means of the forced contact means is
This is placed in the culture vessel 1 and imparts a wave-forming external force to the microbial culture 8 to cause it to move, thereby causing the microbial culture 8 to pass through the porous filtration separation membrane 9 in the gas phase space.
Are forcibly contacted with each other.
A motor (not shown) is eccentrically fixed via a crank, for example, and is fixed to a rotating shaft 11 that reciprocates and is arranged so as to reciprocate in the microorganism culture solution 8. The wing plate 10 of this embodiment is supported by bearings 12 installed on both ends or one of the closed side plates of the culture container 1. Of the rotation shaft 11
The height (vertical length) fixed to the base is slightly shorter than the radius of the cylindrical body, and the reciprocating motion effectively presses the microbial culture solution 8 and effectively performs the oscillating wave-making.
The wave of the microorganism culture solution 8 is raised and dropped along the inner wall surface of the cylinder of the culture vessel 1 so as to jump up into the gas phase space 7, and is forced into contact with the porous filtration separation membrane 9.

At this time, the wing plate 10 in the wave-making means of the present embodiment is arranged such that a pair thereof is opposed to the rotating shaft 11 so as to form a predetermined angle, for example, an angle of 60 degrees, and the rotating shaft 11 is The moving angle, that is, the turning angle of the blade 10 is a predetermined angle,
For example, the angle is set to about 60 to 80 degrees, so that the wing plate 10 divides the microorganism culture solution 8 into three sections, and the microorganism culture solution 8 is made to wave in both directions inside the cylindrical body of the culture vessel 1. Thus, the microbial culture solution 8 is forcibly applied to the porous filtration / separation membrane 9 in the cross direction of the cylinder in the cylindrical body of the culture vessel 1 from opposite directions in accordance with the reciprocating motion of the wing plate 10. At this time, the wing plate 10 is provided with a rotating shaft as an arrangement of the porous filtration / separation membrane 9, and in particular, as a wave-forming means for each of the plurality of blocks or groups of the plurality of parallel stages. In this embodiment, the pair of wing plates 10 are mounted on the same rotating shaft 1.
In contrast, the culture vessel 1 uses two to four pieces so as to be juxtaposed in the longitudinal direction so as to perform oscillating wave making over the entirety of the culture vessel.

In the figure, reference numeral 2 denotes a visual window of the culture vessel 1, reference numeral 4 denotes a tube or pipe for circulating a liquid solvent filtered from the filtrate tank by a pump (not shown) to the culture vessel, and reference numeral 5 denotes a pipe.
For example, air is supplied for culturing microorganisms of aerobic bacteria, and the gas phase space 7 of the culture vessel 1 is discharged so as to discharge carbon dioxide gas.
In the figure, reference numeral 6 denotes a suction / discharge hole for performing gas phase management, and reference numeral 6 denotes a jacket for circulating a temperature-regulating water for controlling the temperature of the culture vessel 1.

The liquid solvent separating device B in the microorganism culturing device A configured as described above is used to separate a liquid solvent from the microorganism culturing solution 8 in the culture vessel 1, and this is separated by the liquid solvent separating method. To explain,
The microbial culture solution 8 contained in the culture container 1 is forcibly brought into contact with a porous filtration separation membrane 9 arranged in the upper gas phase space 7 in the culture container 1, and the liquid solvent in the microbial culture solution 8 is removed from the porous medium. Separation and recovery are performed by absorption and passage of the capillary phenomenon in the filtration and separation membrane 9. In this case, the separation and recovery is performed by replacing the porous filtration and separation membrane 9 in the gas phase space 7 with a hollow pipe-like shape in the present embodiment. The separation is performed by continuously or intermittently, particularly intermittently, applying a pressure to the hollow portion of the separation membrane.

That is, the culture vessel 1 contains a microorganism culture solution 8 in which microorganisms and additives are added and mixed in a culture medium such that a horizontal line passing through the center of the cylinder is a liquid surface. In order to separate the liquid solvent, the wave forming means is operated. In this example, the motor is energized and the rotating shaft 11 is rotated, whereby the wing plate 10 of the wave forming means is reciprocated and the microorganism culture solution 8 is rotated. The microbial culture 8 is dropped on the porous filtration / separation membrane 9, and is forced into contact with the porous filtration / separation membrane 9. The liquid solvent permeated by capillary action is separated into the hollow part by the capillary action of No. 9, and the liquid solvent permeated by the capillary action drops naturally into the hollow part due to gravity. In the example, porous filtration by the above pump Since the hollow portion of the separation membrane 9 is depressurized and depressurized against the surface, it acts to suck the liquid solvent that has penetrated by capillary action, promotes the capillary action, and separates the liquid solvent. It is made sure that it can be done effectively.

On the other hand, microorganisms such as various bacteria remain in the microorganism culture solution 8 in relation to the membrane pore size. However, the clogging substances in the microorganism culture solution 8 such as solid fine particles and insoluble powder are as described above. The forced contact simultaneously contacts and adheres to the surface of the porous filtration and separation membrane 9, but is washed away from the surface together with the microorganism culture by the impact given to the porous filtration and separation membrane 9 by the forced contact, and As a result, the clogging substance is caused to adhere to the surface of the porous filtration separation membrane 9 as much as possible. Thus, the filtration efficiency by the separation of the liquid solvent can be maintained extremely well.

The liquid solvent in the hollow portion, which is separated from the microorganism culture solution 8 by filtration through the porous filtration separation membrane 9, is pulled by the pressure means and collected in the filtrate tank via the tube or pipe 3. After necessary adjustment, it is returned to the microorganism culture solution 8 again so that an environment suitable for culturing microorganisms can be formed.

FIGS. 3 and 4 show another example. In the liquid solvent separator B in the microorganism culturing apparatus A of the present embodiment, the culture vessel 1 is formed in a horizontally-long rectangular box shape, and the upper part is also a gas phase. The space 7 is made of, for example, a horizontally long box made of stainless steel, and the porous filtration separation membrane 9 is separated above the liquid surface of the gas phase space 7 so as to cover it.
The separation membrane 9 of this embodiment is detachably mounted in the longitudinal direction of the box. The separation membrane 9 of the present example is arranged in parallel with a plurality of the separation membranes 9 at a plurality of both side positions slightly separated from the center so that the separation membranes 9 are substantially flat. In this example, for example, five arrangements are used on both sides to ensure the efficiency of the forced contact described above.

In the present embodiment, the forced contact means is formed by a spray forming means for ejecting or scattering the microorganism culture solution 8 in the culture vessel 1 into the gas phase space 7, that is, the spray forming means of the present embodiment. Is above the gas phase space 7 from the intersection of the diagonal lines of the culture vessel 1 through the space separated by the porous filtration separation membrane 9 by the culture solution ejection pump 13 installed in the culture vessel 1 or outside the culture vessel 1. Spout the microbial culture 8
The flat porous filtration membrane 9 is brought into contact with the ceiling surface of the culture vessel 1, in this example, the diffuser 14 provided therein.
By dropping the microbial culture solution 8 in the form of droplets,
The forced contact is made.

The rest of the configuration is basically the same as that of the above-mentioned example, so the same reference numerals are given and the explanation is omitted. In this example, however, in the example of FIG. 1 and FIG. The figure shows a supply tank 15, a pump 16, an air compressor 17, a sterilization filter 18, a filtrate tank 19, and a pump 20 thereof for a microorganism culture solution (substrate solution) for the culture vessel 1 not shown. In the examples of FIGS. 1 and 2, the microorganism culturing apparatus A may be configured according to this.

In the liquid solvent separator B of the microorganism culturing apparatus A of this embodiment, the microorganism culturing solution 8 is also accommodated so that a horizontal line passing through the upper and lower middle positions of the culture vessel 1 is used as the liquid surface, and the liquid solvent is separated. In this example, the droplet forming means is operated, and in this example, the culture solution ejection pump 13 is operated to eject the microorganism culture solution 8 and force the microorganism culture solution 8 into contact with the porous filtration separation membrane 9 by the above-mentioned dropping. Similarly, the liquid solvent is separated and recovered by capillary action of the porous filtration / separation membrane 9 under a reduced pressure state, and the porous filtration / separation membrane 9 for clogged substances is automatically washed by the forced contact of the droplets.
In the same manner, it is possible to prevent the occurrence of clogging, to maintain the filtration efficiency extremely satisfactorily, and to exhibit the same operation and effect as in the above example.

Although the illustrated example is as described above, the porous filtration / separation membrane is formed as a flat panel, which is used together with an integral or separate saucer disposed thereunder.
Separating the liquid solvent in the pan and collecting it in the filtrate tank via the pan, when using forced contact means,
This is to be performed by means of a culture vessel swinging means for mounting and fixing the culture vessel, and a horizontal drop plate provided in the culture vessel and a rotary dropper provided with an abutment plate for guiding and dropping the rotated microorganism culture solution upward. When the forced contact means is wave-forming means, it promotes contact of the microbial culture solution with the porous filtration separation membrane, a diffuser is arranged on the inner wall of the culture vessel, and the forced contact means is splashed. When the pump is used as the forming means, the pump is installed outside the culture vessel, and the pump is configured as a circulating system in which the microorganism culture solution is scattered as droplets from above the gas phase space of the culture vessel by a tube or a pipe. In the practice of the present invention, including, for example, continuously reducing the pressure in the hollow portion of the porous filtration separation membrane, the culture vessel, its gas phase space, the porous filtration separation membrane, Specific materials, shapes, structures, dimensions, capacities, numbers, sizes, uses, etc. of the culture solution, forced contact means used as necessary, wave forming means, droplet forming means, and pressure reducing means of the examples are as follows. Various modifications can be made to an appropriate form without departing from the gist of the invention.

[0023]

Example 1 A culture vessel 1 in a microorganism culturing apparatus A shown in FIGS. 1 and 2 was used for a capacity of 4 L, and yeast extract 10 g / L,
PH containing 30 g / L of tryptone and 5 g / L of glucose
E. coli was inoculated into 2 L of medium No. 7 and cultured at 37 ° C. for 1 day. A substrate solution of the same pH 7 containing 10 g / L of yeast extract and 30 g / L of tryptone was supplied, and the mixture was separated by filtration under reduced pressure. Culture was performed. During this time, the wing plate 10 was reciprocated at a cycle of 60 times per minute to continuously forcibly contact the microbial culture solution 8 with the porous filtration separation membrane 9.
By the day, the cell concentration was 50 g / L, and a viable cell rate of 95% was obtained. At this time, the filtration flow rate was initially 90%, and the dilution rate was 0.3.
h- ¹ , no residual glucose was detected, and the isolation rate of the cells was 99%.

[0024]

COMPARATIVE EXAMPLE 1 Using the same microorganism culturing apparatus A from which the wing plate 10 of the wave-making means was removed, the porous filtration separation membrane 9 was immersed and installed in the microorganism culture solution 8, and the filtration separation was performed under reduced pressure. Was cultured. The filtration flow rate rapidly decreased, and the dilution ratio on the third day of culture was 0.05 h ^-1 . The viable cell ratio was 10% when the cell concentration was 15 g / L.

[0025]

Example 2 Porous filtration of a microorganism culture solution 8 in which the culture vessel 1 in the microorganism culture apparatus A shown in FIGS. 3 and 4 is used for a capacity of 5 L and the ejection of the microorganism culture solution 8 by the culture solution ejection pump 13 is set to 3 L per minute. Culture was performed under the same conditions as in Example 1 except that forced contact with the separation membrane 9 was continued.
By 3 days of culture, the cell concentration was adjusted to 60 g / L, and the viable cell rate was 90%.
%. At this time, the filtration flow rate was similarly 90% at the initial stage, the dilution rate was 0.3 h ^-1 , no residual glucose was detected, and the bacterial cell separation rate was 99%.

[0026]

[Comparative Example 2] Using the same microorganism culturing apparatus A without operating the culture liquid ejection pump 13, the porous filtration / separation membrane 9 was immersed in the microorganism culture liquid 8, and filtered and separated under reduced pressure. Was cultured. The filtration flow rate rapidly decreased, and the dilution rate on the third day of culture was 0.05 h ^-1 , and the cell concentration was 10 h.
The viable cell ratio was 8% in g / L.

[0027]

The present invention is configured as described above.
According to the first aspect of the present invention, there is provided a microorganism capable of performing filtration of a porous filtration separation membrane as satisfactorily as possible, eliminating clogging caused by the filtration, and maintaining filtration efficiency as satisfactorily as possible. It is possible to provide a method for separating a liquid solvent in a culture device, and the invention according to claim 2 provides, in addition to the above,
Promote the filtration of liquid solvent by the porous filtration separation membrane,
A sufficient filtration performance can be ensured, and the invention according to claim 3 performs filtration of the porous filtration separation membrane as satisfactorily as possible and eliminates clogging accompanying the filtration. It is possible to provide a liquid solvent separation device in a microorganism culturing device capable of maintaining the filtration efficiency as good as possible. The forced contact of the microorganism culture solution with the filtration / separation device can be performed easily and reliably as a preferred forced contact means. It is possible to promote the filtration of the solvent and ensure sufficient filtration performance.

[Brief description of the drawings]

FIG. 1 is a longitudinal sectional view of a microorganism culturing apparatus at an intermediate position of a culture vessel in a direction crossing a cylinder.

FIG. 2 is a longitudinal sectional view of the microorganism culturing apparatus at an intermediate position in the front-rear direction of the culture vessel.

FIG. 3 is a partially cutaway perspective view of a microorganism culturing apparatus of another example.

4 is a longitudinal sectional view of the microorganism culturing apparatus of FIG. 3 at an intermediate position in the front-rear direction of the culture vessel.

[Explanation of symbols]

 Reference Signs List A Microorganism culture device B Liquid solvent separation device 1 Culture vessel 7 Gas phase space 8 Microorganism culture solution 9 Porous filtration separation membrane 10 Blade plate 13 Jet pump

──────────────────────────────────────────────────続 き Continuation of the front page (72) Inventor Yukio Sakaba 1-36-2 Hakusan, Bunkyo-ku, Tokyo Lions Mansion Bunkyo Hakusan 108 Within Geo Support Co., Ltd. (56) References JP-A-6-98758 (JP, A) JP-A-5-95778 (JP, A) JP-A-5-84429 (JP, A) (58) Fields investigated (Int. Cl. ⁶ , DB name) C12M 1/00-3 / 04 B01D 61/00-71/82

Claims (6)

(57) [Claims] 1. A microbial culture solution contained in a culture container is forcibly brought into contact with a porous filtration separation membrane disposed in an upper gas phase space in a culture container, and the liquid solvent in the microorganism culture solution is subjected to the porous filtration. A method for separating a liquid solvent in a microorganism culturing apparatus, wherein separation and recovery are performed by absorption and passage of capillary action in a separation membrane. 2. The method according to claim 1, wherein the porous filtration separation membrane in the gas phase space is a hollow pipe-shaped separation membrane, and the hollow portion is continuously or intermittently brought into a pressure-reducing state. A liquid solvent separation method in a microorganism culturing apparatus. 3. A culture vessel, a porous filtration separation membrane disposed in a gas phase space above the culture vessel, and a microorganism culture solution contained in the culture vessel being forcibly applied to the porous filtration separation membrane in the gas phase space. A liquid solvent separation device in a microorganism culturing device, comprising: forced contact means for contacting the liquid solvent. 4. The apparatus for separating a liquid solvent in a microorganism culturing apparatus according to claim 1, wherein said forced contact means is constituted by wave-forming means for wave-forming and oscillating a microorganism culture solution in a culture vessel. . 5. The liquid in the microorganism culturing apparatus according to claim 1, wherein said forced contact means is formed by a droplet forming means for ejecting or scattering a microorganism culture solution in a culture vessel into a gas phase space. Solvent separator. 6. The method according to claim 1, wherein the porous filtration separation membrane is formed as a hollow pipe-shaped separation membrane and further provided with a pressure reducing means for continuously or intermittently reducing the pressure of the hollow portion. A liquid solvent separation device in the microorganism culturing device according to claim 3, 4 or 5.