JP2774853B2 - Cleaning and sterilization reactor for immobilized substances - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION (Industrial application field) The present invention relates to an apparatus for producing a useful substance using an immobilized biocatalyst such as an enzyme or a microorganism as an immobilized substance without deactivating the immobilized substance. The present invention relates to a high-efficiency washing / sterilization reaction apparatus for immobilized substances that can be washed / sterilized and used repeatedly.

(Prior Art) Conventionally, a variety of technologies for producing useful substances using immobilized biocatalysts such as enzymes and microorganisms have been proposed, including packed bed type, fluidized bed type, stirred bed type, and membrane type.

An apparatus for cleaning and sterilizing an immobilized biocatalyst in these reactors is disclosed in Japanese Patent Publication No. 62-33873. In which the immobilized biocatalyst dispersed in the carrier can be circulated and transported.

In the techniques described above, a surfactant such as a quaternary ammonium salt, propylene glycol at 10 to 50%, or the like is used as a bactericide for the immobilized biocatalyst.

Also, in JP-A-1-174375, as a reactor with a fluidized tank, the immobilized biocatalyst is washed and sterilized in a fluidized tank, and when extracting useful substances after the reaction, it is performed in a lower column. Has become.

Furthermore, Japanese Patent Application Laid-Open No. 1-2228454 has been proposed as an improvement of the above-mentioned Japanese Patent Application Laid-Open No. 1-17375, in which the immobilized biocatalyst is sterilized by ultraviolet rays.

(Problems to be Solved by the Invention) Among the conventional techniques described above, in the sterilization method disclosed in Japanese Patent Application Laid-Open No. 62-33873, a decrease in the activity of the immobilized enzyme is unavoidable. When the concentration of the agent is reduced, there is a problem that the bactericidal effect is reduced.

In the reactor with a fluidized tank disclosed in JP-A-1-174375, washing of the immobilized biocatalyst and washing of the column layer are performed at the same time, and sterilization is performed at the same time. When sterilizing mainly the immobilized biocatalyst, there is a problem that sterilization of the device is insufficient.

Further, in the device disclosed in Japanese Patent Application Laid-Open No. 1-228454, wherein sterilization of the immobilized biocatalyst is performed under ultraviolet light under a certain condition, irradiation of the immobilized biocatalyst is carried out in order to uniformly irradiate the ultraviolet light. There is a problem that the state is not stable.

In particular, sterilization using ultraviolet light is performed under the condition that ultraviolet light irradiation is applied stably and uniformly to the entire surface of the immobilized biocatalyst, and irradiation on a flat body is desirable if possible.

When dispersing the immobilized biocatalyst, a device such as a stirrer is damaged by mechanical stress, and the immobilized biocatalyst is damaged, which is not good.

In view of the above conventional problems, the present invention disperses an immobilized biocatalyst without breaking it, and an immobilized substance capable of performing an averaged ultraviolet irradiation on the immobilized biocatalyst without deactivating the immobilized biocatalyst. It is an object of the present invention to obtain a washing and disinfecting reaction device.

(Means for Solving the Problems) In order to achieve the above object, the present invention has an object to provide the following apparatus for washing and sterilizing an immobilized substance.

In other words, there is provided an apparatus for cleaning and sterilizing an immobilized substance, wherein a holder for holding an immobilized substance in a cleaning and sterilizing reaction tank repeatedly rotates and / or moves up and down, and irradiates ultraviolet rays from above the apparatus.

In addition, when the operation range of raising and lowering the holding body of the immobilized substance in the washing and sterilizing reaction tank is performed in a range where the holding body in the reaction tank smoothly contacts the outer body during the washing and sterilizing reaction, and when the washing and sterilizing reaction is completed. A cleaning and disinfection of the immobilizing substance, which is configured to be able to be lowered to a range where the holding body does not come into contact with the outer body and to discharge the immobilizing substance to a lower portion of the outer body, and perform a two-stage lifting operation to return to the original position after discharging. It is a reactor.

Further, there is provided an apparatus for cleaning and sterilizing an immobilized substance, in which one or more columns are provided around the reactor for washing and sterilizing the substance for immobilizing, the column containing the immobilized substance and extracting a useful substance by an extract.

The column is a cylindrical body made of a support for the immobilized substance, and a conduit for introducing the extract is connected to the upper part of the column, and the column is raised and lowered by raising and lowering the conduit to fix the column when it is at the upper part. The immobilized substance is discharged from the column into the washing and disinfection reaction tank, and when it is at the bottom, the immobilized substance is injected into the column from the washing and disinfection reaction tank, and then the immobilized substance is washed and disinfected by the extract to elute useful substances. Yes,
UV irradiation is performed at an irradiation intensity of 0.5 mW / cm ² or less.

(Function) In the case of the present invention, the column is raised and lowered to discharge the immobilized substance from the column to the washing and sterilizing reaction tank when it is at the top, and to inject the immobilized substance from the washing and sterilizing reaction tank to the column when it is at the bottom. Then, the useful substance is eluted by the extract, and the immobilized substance in the washing and disinfecting reaction tank is subjected to a rotation operation and a lifting / lowering operation by the holding body so that the ultraviolet irradiation can be uniformly received.

(Example) Hereinafter, an example illustrated in the drawings will be described.

(1) is an exterior body of the washing and disinfecting reaction device (A), in which a top plate has air bleeding (2), and an inner surface of the top plate has an ultraviolet germicidal lamp (3). A plurality of the germicidal lamps (3) may be arranged according to the situation. Retaining body guide band (4) on the inner surface of the trunk of the exterior body (1)
The holder (6) can be moved up and down along the guide groove (5) of the holder guide band (4).

The holding member may be a plate member provided with a slit through which the immobilization carrier does not pass, a mesh body through which the immobilization carrier does not pass, or a combination of both.

The holding body (6) moves up and down between the cam groove (8) of the rotating body (7) and the guide groove (5). That is, the cam pin (9) of the holder (6) is fitted into the cam groove (8),
The guide pin (10) of the holding body (6) fits into the guide groove (5), and moves up and down by rotation of the rotating body (7).

The rotating body (7) is composed of a shaft (12),
The rotator (7) is driven through gears (13) and (14), and moves up and down from the cylinder (15) via a lifting shaft (15a).

To enable the above operation, the gear (14) is a long gear that can move up and down while meshing with the gear (13), and the guide pin (10) of the holder (6) is guided by the guide. A guide groove (5) is extended to the skirt portion (16) of the holder guide band (4) so as not to slip out of the groove (5),
The guide (4) other than the skirt (16) has an opening (1).
7) You are.

Therefore, when the holder (6) is lowered to the skirt portion (16) by the cylinder (15), the holder (6)
Will be facing the opening (17).

The rotating body (7) is driven to rotate by a rotating shaft (19) from a gear (14), and is connected to a lifting shaft (15a) by a universal joint (18).

Therefore, even if the rotating shaft (19) rotates, the lifting shaft (15
a) does not rotate.

The rotating body (7) may be moved up and down by a cylinder as shown in the figure, but may be moved up and down by the rotation of the inclined disk so that the axis of the holder comes into contact with the disk, or may be moved up and down by a cam mechanism. You may do it. Although the holder shown in FIG. 1 moves up and down, the holder may rotate or may rotate and move up and down simultaneously. in this case,
As shown in the figure, it is not necessary to convert the rotational force into the elevating operation of the holder, and the holder may be directly connected to the driving device via a shaft. Further, in the case of the holder, if the surface of the holder is made uneven or wavy and slits are provided, it is effective in dispersing the immobilizing substance.

A discharge control body (20) is rotatably arranged on the inner surface of the bottom of the exterior body (1) of the washing and sterilization reaction tank by a gear device (not shown), and a discharge port opened to the discharge control body (20). (21) and (22) are connected to the discharge column (23) or drain drain (24) of the target column to discharge the immobilized carrier and the processing solution. In this case, the discharge control body (20) is provided with the holder guide band (4).
It is raised up to a position higher than the opening (17) other than the skirt portion (16) and is connected to the gear device, so that the fixed carrier is not damaged.

A plurality of column outer cylinders (25) are disposed around the outer body (1) of the washing and disinfecting reaction tank, and an extract solution conduit (27) is provided therein.
Moves the column (26) up and down. The bottom of the column (26) is a mesh (29), and the upper part of the column outer cylinder (25) has an air vent (28) and a discharge port (30) with a cock at the bottom.

The column may be raised and lowered by raising and lowering the extract liquid conduit (27) as shown in the figure. However, the column is not connected to the extract liquid conduit (27) but is moved up and down by a separate raising and lowering mechanism using the upper and lower columns as a holding network for immobilized carriers. It doesn't matter.

Specifically, the column may be raised and lowered from the lower part of the column outer cylinder (25) by a cylinder, or a screw groove may be provided upright at the center to form a fitting part with the column with a screw, and the screw rod may be rotated. It may be raised and lowered by letting it go.

Further, in addition to the above-mentioned discharge pipe (23) leading to the bottom of the outer casing (1), the column outer cylinder (25) is connected to the outer casing (1) from the upper portion of the holder guide band (4) of the outer casing (1). Connected inlet pipe (3
1) There is.

Furthermore, the discharge pipe (23) and the inlet pipe (31) serve as an inlet / outlet for the solidified carrier of the column, but when not connected to the inlet / outlet (21) of the outer body of the washing / sterilizing reaction tank, the discharge control body ( 20)
Separately, it is preferable to rotate the column (26) itself and raise and lower the port (21a) without connecting the port (21a) to the port (21) of the washing and disinfecting reaction tank (1).

The opening and closing of the discharge pipe (23) and the inlet pipe (31) may be performed by rotating the discharge control body (20) or the column itself, or by a valve. Since it is not preferable that the carrier remains, it is preferable that the bottom of the tank (1) and the valve surface be made smooth by a ball valve, or the bottom of the tank is made smooth by a Y valve.

The holding body (6) is in contact with the exterior body (1) of the tank (A) smoothly. However, since the guide groove (5) is provided so as not to rotate as described above, the guide groove (5) is provided on the holding body (6). Skirt (16) extending to the lower end where (6) descends
Is provided as shown, but when the holder (6) itself is rotated, it is performed separately from the elevating operation, and in this case, the guide groove is not necessary.

Now, when proteins and the like adhere to the immobilized biocatalyst and the pressure loss is so severe that it is desired to wash and sterilize the immobilized biocatalyst, the cleaning liquid and the liquid for sterilization are supplied from the liquid inlet (31) of the outer body (1) to the outer body ( 1) filled in the lower part of the holder (6),
It is filled up to the upper part of the holder (6).

Then, the motor (11) is driven to rotate the rotating body (7) from the rotating shaft (19), and the holding body (6) is rotated by the cam groove (8).
Is moved in a range where the holder (6) is in smooth contact with the outer package (1), that is, in a range of the holder guide band (4), and ultraviolet light is irradiated from above by an ultraviolet lamp (3). Is carried out to wash and sterilize the immobilized biocatalyst held by the holder (6). Irradiation intensity of 0.5 mW / cm ³ or less shows a bactericidal effect without inactivation and is appropriate. The reason for this is disclosed in JP-A-1-220454.

The ultraviolet germicidal lamp (3) is provided on the ceiling of the tank, but it is preferable that the ultraviolet germicidal lamp is installed so as to be adjustable in position so that the distance from the holder (6) can be adjusted. (3) is installed. This is because the device can be used in various ways if it can be adjusted according to the amount of liquid at the time of sterilization or the type of immobilized biocatalyst.

To adjust the distance to the holding body (6), specifically, the ceiling and the ceiling are slidably mounted on the inner wall of the tank.

At the time of cleaning and sterilization, it is desirable to perform treatment by a not-shown showering device or the like, and this showering device is preferably provided around the upper portion of the inner wall of the tank so as not to hinder ultraviolet irradiation.

When the cleaning liquid and the sterilizing liquid have been drained from the drain (24), the discharge regulating body (20) is rotated to move the discharge port (22) away from the drain (24).

Outer body of immobilized carrier with immobilized biocatalyst attached (1)
The column (26) is raised by the extract conduit (27), and the port (21a) of the column (26) is made to coincide with the inlet tube (31). 6) The solution is allowed to flow into the upper portion, and the immobilized carrier on the holder (6) is washed while being lifted and lowered. Then, ultraviolet irradiation is performed with an ultraviolet sterilizing lamp (3), and then the reaction is carried out. After cleaning and sterilization, the cleaning liquid and the liquid for sterilization are driven to the discharge control body (20) to discharge the discharge port (22) to the outside by matching the drain port (24) with draining (24). Thereafter, the holding body (6) does not come into contact with the exterior body, that is, it is lowered by the cylinder (15) to the skirt portion (16) of the holding body guide band (4), and the discharge opening (21) is formed by the discharge regulating body (20). ) Coincides with the discharge pipe (23), and then the column (26) is lowered by the extract liquid conduit (27), and the opening (21a) is connected to the discharge pipe (2).
3). Then, the immobilized carrier is injected into the column (26) by the guide liquid from the tank (A), and then the useful substance is eluted from the outlet (30) by the extract. The outlet (30) is closed when the column (26) is at the raised position.

The immobilized carrier in the column (26) from which the useful substance has been discharged is raised again by the column, reduced from the inlet pipe (31) into the tank (1), and raised and lowered by the holder (6) and sterilized by ultraviolet rays. The reaction is carried out by irradiating ultraviolet rays with the lamp (3). In addition, it is desirable that the lower part of the extract liquid conduit (27) of the column (26) is an inverted funnel provided with a slit plate or a mesh so that the extract can flow evenly and evenly in the column (26). The detection of the position of the column, the detection of the liquid level in the tank, the detection of the position of the discharge regulating body, and the like are performed by an automatic control device via a detection device (not shown).

Thus, by arranging a plurality of columns around the washing and disinfecting reaction tank, and performing the washing and disinfecting reaction process by shifting the plurality of columns temporally, it is possible to continuously produce useful substances. Moreover, since the circulation of the immobilized carrier can be carried out in a closed system, it is possible to carry out circulation production without contamination.

In addition, in the present invention, the sterilization and washing reaction of the immobilized carrier and the washing and sterilization of the device can be performed separately. That is,
Washing and sterilization of the device can be performed separately by discharging the immobilized carrier to the column.

(Effects of the Invention) According to the present invention, the washing and disinfecting reaction treatment of the immobilized substance can be performed after the washing and disinfection of the equipment, so that the invention relating to the washing liquid and the disinfection method can be separately carried out, which is efficient.

In addition, since a plurality of columns can be connected to one washing and disinfecting reaction apparatus, processing can be performed with high efficiency.

Furthermore, the stirring operation of the immobilized substance is performed by rotating the holding plate and / or
Or it is soft because of the lifting and lowering operation, there is no destruction of the immobilization substance due to friction between the immobilization substances and the shearing force due to stirring, and it is possible to disperse evenly on a flat surface in sterilization operation and efficient sterilization is possible . In addition, by washing, disinfecting, and reacting multiple columns in time, continuous production of useful substances becomes possible, and circulating production of immobilized substances can be performed in a closed system, so that circulating production without contamination is possible. Is possible, and the effective use of the immobilized substance is possible.

[Brief description of the drawings]

 FIG. 1 is a cutaway view of the device of the present invention, FIG. 2 is a cutaway view of a drive device, and FIG. 3 is an enlarged view of a skirt. (1) ... exterior body of washing and disinfection reaction tank (2) ... air bleeding (3) ... ultraviolet sterilization lamp (4) ... holder guide band (5) ... guide groove (6) ... holder (7) Rotating body (8) Cam groove (20) Discharge regulating body (23) Discharge pipe (24) Drain (25) Column outer cylinder (26) Column (27)… Extract liquid conduit (28)… Vent air (29)… Mesh (30)… Outlet (31)… Inlet pipe

────────────────────────────────────────────────── ─── Continuation of the front page (56) References JP-A-1-228454 (JP, A) JP-A-1-174375 (JP, A) JP-B-62-33873 (JP, B2) (58) Field (Int.Cl. ⁶ , DB name) C12M 1/00-3/10 C12N 11/00

Claims (5)

(57) [Claims] An apparatus for cleaning and sterilizing an immobilized substance, wherein a holder for holding an immobilized substance in a cleaning and sterilizing reaction tank repeatedly rotates and / or moves up and down, and irradiates ultraviolet rays from above. 2. The operation range for raising and lowering the holder of the immobilizing substance in the washing and disinfecting reaction tank is performed in a range where the holding body in the reaction tank smoothly contacts the exterior body during the washing and disinfecting reaction, and the washing and disinfecting reaction is completed. When the holding member is moved down to a range where the holding member does not come into contact with the outer package, the immobilizing substance can be discharged to a lower portion of the outer package. An apparatus for washing and sterilizing an immobilized substance according to claim 1. 3. The method according to claim 1, wherein one or more columns are provided around the reaction vessel for washing and sterilizing the immobilized substance, the column containing the immobilized substance and extracting the useful substance from the extract. A washing and disinfecting reactor for the immobilized substance according to the above. 4. A column at the bottom of the column made of a support for the immobilized substance, a conduit for introducing an extract is connected to the upper part of the column, and the column is raised and lowered to raise and lower the column. (1) discharging the immobilized substance from the column to the washing and disinfecting reaction tank when it is in the column, injecting the immobilized substance from the washing and disinfecting reaction tank into the column when it is below the column, and then eluting the useful substance by the extract. (2) A device for washing and sterilizing an immobilized substance according to (2) or (3). 5. An apparatus for washing and sterilizing an immobilized substance according to claim 1, wherein the ultraviolet irradiation is performed at an irradiation intensity of 0.5 nW / cm ² or less.