JPS6098981A - Bioreactor of rotary disc type - Google Patents

Abstract

PURPOSE:The titled reactor having high activity of reactor, not causing concentration and clogging of immobilized organism catalyst, obtained by equipping a revolving shaft in a tank rectangularly with plural discs with thin layers capable of adsorbing an enzyme or microorganism by ion bonding method in parallel. CONSTITUTION:The rotary shaft 21 supported horizontally in a reactor is rectangularly equipped with the plural rotary discs 25 in parallel approximately in equal distance with the spacers 26 put between them. Immobilization of enzyme, for example, is carried out by immersing previously sheets for adsorbing an enzyme in warm water, swelling fully the sheets, activating them with hydrochloric acid with a given concentration, washing them with water, fixing them to the discs, attaching the discs 25 to the shaft 21, setting the shaft to the reactor. The the reactor is charged with an enzymatic solution, and the enzyme is adsorbed with rotating the shaft 21. The reaction in the prepared bioreactor is carried out by feeding 12 an enzymatic solution to it, and by recovering the reaction solution from the outlet 20. The thin layers supported by the discs 25 are made of a material capable of adsorbing and immobilizing an enzyme, microorganism, etc. by ion bond method, for example, invertase, etc. is immobilized to an anion exchange resin having an anion exchange group specific to the enzyme, the microorganism, etc., respectively.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a reactor in which reactions are carried out using immobilized enzymes or bacteria.

Various bioreactors have been developed in which reactions are carried out using biocatalysts in which enzymes or bacteria are immobilized on a carrier material, but most of them are bioreactors filled with immobilized biocatalysts using bead-shaped carriers. A packed bed format is used.

However, in such a packed bed type bioreactor, there is a problem that the bead-shaped immobilized biocatalyst is compacted in the container, causing clogging, and as a result, the activity decreases rapidly. In addition, if the substrate solution supplied to the reactor has a high viscosity or contains solids, the pressure drop in the packed bed will increase, and channeling will likely occur in the flow of the substrate solution, resulting in reactor efficiency. often decreases.

As a bioreactor that does not cause clogging or pressure loss, an actor using plate-shaped or membrane-shaped immobilized enzymes or bacterial cells has been proposed. Rotating coroot reactors can also be seen (
(For example, 4f r Catalyst J Vol. 21, pp. 445-449,
(1979).

However, all of these rotating disk reactors use a disk on which microbial cells and enzymes are immobilized in advance using a carrier such as collagen or polyacrylamide, which poses problems in terms of strength in large-scale industrial devices. If the activity of the immobilized enzyme or bacterial cells decreases, the disk itself needs to be renewed.

The present invention has high reactor activity, the immobilized biocatalyst does not cause compaction or clogging, and the contact with the substrate solution in the reactor is good, and high reactor activity can be maintained for a long period of time. The present invention provides a rotating disk type bioreactor in which immobilized enzymes or bacterial cells can be easily exchanged.

That is, in the present invention, a plurality of disks each holding a thin layer having the ability to adsorb enzymes or bacteria by an ionic bonding method are arranged in parallel at right angles to the prepacking rotation shaft on a rotation shaft provided in a tank. This is a rotating disk type bioreactor characterized by:

In the present invention, the thin layer held on the rotating disk is made of a material that has the ability to adsorb and immobilize enzymes, bacteria, etc. using an ionic bonding method. Several such materials are already known; for example, invertase and glucose isomerase are each immobilized on anion exchange resins that have specific anion exchange groups1, and lipase is immobilized on cation exchangers that have sulfone groups. known to bind. It is also known that a type of bacteria that has organic matter oxidizing properties is adsorbed and immobilized on anion exchange resin Dowex-1. In the present invention, such an ion exchanger can be used as a thin film or as a thin layer laminated on a disk. Especially JP-A-55-45802
Bonded fibers made from aminoacetalized polyvinyl alcohol, such as those disclosed in No. It can be used as

Various structures can be considered for the rotating disk in which the ion-binding thin layer is held on the disk, but for example, as shown in FIG. The rotary disk can be constructed by fixing with other suitable methods. The rotating disk is provided with a hole 6 through which the rotating shaft passes. As another method, the rotating disk may be constructed by directly stacking and fixing fibers, particles, membranes, etc. having ionic binding properties on the disk 1.

FIG. 2 shows another embodiment of the rotating disk of the present invention, in which the ionic bonding thin layer 4 is sandwiched between mesh-like disks 5 to constitute the rotating disk, and FIG. As shown in FIG. 2, it is also possible to construct a structure in which the ion-bonding thin layer 4 is pressed and held on both sides of the disk 1 by mesh-like disks 5, respectively.

In the bioreactor of the present invention, rotating disks as described above are arranged in parallel on a rotating shaft and installed in a reaction tank, and the substrate solution introduced into the tank comes into contact with the immobilized biocatalyst on the rotating disk. The size and number of disks are arbitrary.
Further, the rotation axis can also be provided horizontally, vertically, or inclined depending on the case.

Figure 4 shows an example of the bioreactor of the present invention using such a rotating disk. FIG.

A cylindrical glass body 16 having a supply port 12 into which the raw material liquid 1.1 is introduced is closed on both sides by stainless steel side plates 15 and 16 via a rubber gasket 14 to form a watertight reactor. Side plates 15 and 16 are tightening rods 17
and is tightened to the body 16 by a nut 18. Further, the side plate 16 is provided with an outlet 20 for taking out the reaction liquid.

Inside the reactor, a rotating shaft 21 has Teflon bearings 2 at both ends.
2, it is rotatably supported horizontally by the rotating shaft 2.
A rotating magnet 23 is fixed to a rotating shaft at one end of the reactor 1, and this rotating magnet 23 is guided by the rotation of another rotating magnet 24 provided outside the reactor and rotates, causing the rotating shaft 21 to rotate. Empower. In other parts of the rotating shaft 21, a plurality of rotating disks 25 are arranged perpendicularly to the rotating shaft 21 with a spacer 26.
They are arranged in parallel at approximately equal intervals.

To immobilize the enzyme, for example, heat the enzyme adsorbent sheet to 80°C in advance.
After soaking in hot water for 1 hour to sufficiently swell, I
After immersing it in N-H (!l solution for 2 hours to activate it, and then thoroughly washing it with water, attach it to a disk as a rotating disk, attach it to the rotating shaft, and install it in the reactor. The enzyme solution is filled in the reactor, and the enzyme is adsorbed while rotating the rotary shaft inside the reactor by rotating the external rotating magnet.

It is usually desirable to take an adsorption time of 1 hour or more for adsorption. If deformation of the thin layer due to swelling is prevented,
It is also possible to perform all of the above-mentioned treatments such as activation within the reactor.

The reaction in the bioreactor of the present invention in which the enzyme is immobilized is carried out by supplying the substrate solution from the supply port and recovering the reaction solution from the discharge port, but the reaction temperature, rotation speed of the disc, The optimum conditions for the residence time of the reaction solution, etc. are determined empirically based on the amount of immobilized enzyme, the concentration of the substrate solution, etc.

If the activity of the immobilized enzyme decreases due to the continuation of the reaction, the adsorbed enzyme is desorbed using, for example, a 5%-NaOt aqueous solution, then activated again with a 1N-HCl aqueous solution, and then rotated again with a new enzyme solution. The reactor can be reactivated by adsorbing the enzyme onto the ion exchanger disk.

By changing the type of ion exchanger attached to the rotating disk, such as ion exchange fiber or ion exchange resin, a variety of enzymes such as invertase, gelcose isomerase, lipase, aminoacylase, and catalase can be immobilized. Also, some types of ion exchange resins can adsorb bacteria through their ionic binding force.

Example Using an apparatus similar to the reactor shown in FIG. 4, the enzyme invertase was adsorbed and immobilized on an enzyme adsorption sheet.

The reactor has a cylindrical shape with an inner diameter of 30 mm, and the actual volume (work
Using a material with a dry weight of 25 mt and a thickness of Q of 5 mm, a disk with a dry thickness of J5 mm and a diameter of 28 mm was placed on both sides of a hard polyvinyl chloride disk with a diameter of 28 mm.
An enzyme adsorption sheet (non-woven fabric made of amimeacetalized polyvinyl alcohol composite fiber manufactured by Nichibi Co., Ltd.) of 1.5 mm in size was swollen in dry water at 80°C in advance, and then soaked in lN-HCl at 30°C.
It was activated by immersing it in an aqueous solution for 2 hours, thoroughly washed with water, and then fixed with staples and used as a rotating disk.

An aqueous solution of invertase (manufactured by Toyobo Co., Ltd., l 23 u/mg) was placed in a reactor equipped with a rotating disk as shown in FIG. 4, and the rotating disk was heated at 42 r, p, m for 2 hours at 30°C.

Enzyme adsorption was performed by rotating the tube.

Next, an enzymatic conversion reaction was carried out using a 20% by weight sucrose solution as a raw material liquid. For the reaction, the raw material liquid storage tank and reactor are
It was placed in a constant temperature water bath at 5°C.

The number of rotating disks installed in parallel in the reactor is 10 (the number of sheets is 20).
The experiment was conducted for the case of SV2, SV2, 5, and SV5 of the flow rate of the sucrose solution. The conversion rate of the sucrose solution by invertase was as shown in FIG. 5 when the rotational speed of the rotating disk was varied within 48 hours after immobilization of invertase. As is clear from the results shown in the figure, rotation of the disk significantly improves the conversion rate.

When using the same device with 10 disks (20 sheets), the amount of enzyme adsorption per reactor volume is 2200 U/crn.
3.5V==2.5, reaction temperature 55°C1 disk rotation speed 4
The change in conversion rate when a 20 wt % sucrose solution was continuously treated at 2 r, p, m was as shown in FIG.

[Brief explanation of drawings]

FIGS. 1, 2, and 3 are exploded views showing the structure of a rotating disk used in the present invention, and FIG. 4 is a sectional view showing one mini-reactor according to an embodiment of the present invention. FIGS. 5 and 6 are graphs showing the conversion rate of a sucrose solution conversion reaction using an immobilized invertase using a minireactor. The correspondence between the main parts illustrated and the symbols is as follows. DESCRIPTION OF SYMBOLS 1... Disc, 2... Fibrous sheet, 4... Ion adsorbent thin layer, 5... Reticular disk, 12... Supply port, 13... Body, 14... Packing , 15.1
6... Side plate, 20... Discharge port, 21... Rotating shaft, 23.24... Magnet rotating body, 25... Rotating disk, 26:... Spacer. Applicant: Mitsubishi Kakoki Co., Ltd. Agent: Katsura Atsuta, Patent Attorney

Claims (4)

[Claims] (1) A plurality of disks holding a thin layer capable of adsorbing enzymes or bacteria using an ionic bonding method are arranged in parallel on a rotating shaft provided in the tank at right angles to the rotating shaft. Rotating disk type % type % characterized by (2) A sheet in which the thin layer is made of ion-binding fibers;
A bioreactor according to claim (1). (3) The bioreactor according to claim (1), wherein the thin layer is a granular or fibrous ion exchanger laminated and fixed on a disk. (4) The bioreactor according to any one of claims (1) to (3), wherein the thin layer is sandwiched by reticulated discs.