JPS6343920Y2 - - Google Patents

Description

[Detailed Description of the Invention] The present invention relates to an enzyme reaction device, and more particularly to an enzyme reaction device that uses water-dispersed polymer particles containing magnetic powder and on which an enzyme is immobilized as an immobilized enzyme.

In recent years, immobilized enzymes in which enzymes are immobilized on water-insoluble carriers have come to be used in the same way as solid catalysts, but in many cases, these conventional immobilized enzymes are made of particles with a particle size of several mm. It is usually used by being filled in a reaction container. However, such a packed bed of particulate immobilized enzymes easily becomes clogged when a highly viscous substrate or macromolecule is passed through it, and diffusion of the substrate is inhibited.

To solve these problems, immobilized enzymes in which enzymes are immobilized on water-dispersed polymer particles have already been proposed (for example, Japanese Patent Application Laid-Open No. 57-150386).
According to such immobilized enzymes, the enzymatic reaction with the substrate can be carried out in a dispersed system, and therefore,
Immobilized enzymes can move within the reaction system in the same way as free enzymes, so unlike the conventional case where immobilized enzymes are immobilized within the reaction system, the substrates can easily diffuse even if the substrate has a high molecular weight. It has the advantage that the enzymatic reaction can be carried out quickly.

On the other hand, when carrying out enzymatic reactions using such aqueous dispersions of immobilized enzymes, it is necessary to separate them from the immobilized enzymes, reaction products, and unreacted substrates, especially when carrying out enzyme reactions continuously. Yes, in this case,
Although it is practically advantageous to use a separation membrane, there is a problem in that the particle size of the immobilized enzyme is small, so that the separation membrane is easily clogged with the immobilized enzyme.

The present invention was made to solve the above-mentioned problems, and when performing an enzyme reaction using an immobilized enzyme in which the enzyme is immobilized on water-dispersed polymer particles, It is an object of the present invention to provide an enzyme reaction device that allows easy separation.

The enzyme reaction device according to the present invention includes (a) a cylindrical container, (b) a rotating shaft provided in the axial direction of the container, (c) a stirring blade that can rotate together with the rotating shaft, and (d) the above-mentioned components. and a magnet attached to the wall of the container or the stirring blade, and in the container, while being stirred by the stirring blade, the particles containing the magnetic powder and having the enzyme immobilized thereon are produced with a particle size of 0.03~
It is characterized by reacting an aqueous dispersion of 2μ water-dispersed polymer particles with a substrate.

The present invention will be described below based on drawings showing embodiments.

FIG. 1 shows an embodiment of the enzyme reaction apparatus according to the present invention, in which magnets 4 are arranged in multiple stages in an annular manner along the inner wall surface of a cylindrical container 3 having a substrate solution inlet 1 and a reaction mixture outlet 2. It is arranged. A rotary shaft 5 is rotatably supported in the axial direction of the container, and stirring blades 6 are protruded from the rotary shaft radially in the radial direction so that their tips are close to the magnet body. ing.

In the enzyme reaction apparatus of the present invention, an aqueous dispersion of water-dispersible polymer particles containing magnetic powder and immobilized enzyme is present in the container. Preferably, a separation membrane 7 that does not allow the above-mentioned immobilized enzyme to pass through but allows the reaction mixture to pass through is provided at the substrate solution inlet and reaction mixture outlet.

The immobilized enzyme used in the present invention is, for example,
By emulsion polymerizing the required monomers in an aqueous system in which magnetic powder is present in colloidal form, water-dispersed polymer particles containing magnetic powder are obtained. The enzyme can be obtained by immobilizing the enzyme by any method. Such water-dispersed polymer particles have an average particle diameter of 0.03
-2μ, preferably 0.1-1μ. If the average particle size is too small, it is necessary to use a separation membrane with a small micropore size as described above, and as a result, the flow rate of the reaction mixture through the membrane becomes low, and it takes a long time to obtain the reaction product. It will be necessary to
On the other hand, if it is too large, the dispersibility of the immobilized enzyme will be poor, and the particle surface area per unit volume of the particles will be small, which is not preferable because the reaction activity of the immobilized enzyme will be relatively small.

According to the apparatus of the present invention, the substrate solution is supplied into the container from the substrate solution inlet, and the substrate and the immobilized enzyme are brought into contact with each other under agitation caused by the rotation of the rotating shaft to perform an enzyme reaction. Here, since the polymer particles used as carriers of the immobilized enzyme are water-dispersed polymer particles and contain magnetic powder, they are attracted by the magnet while the rotating shaft is rotating. However, unlike the substrate and reaction products, which are not attracted by the magnet, they are stirred by the stirring blade.
While remaining substantially within the container, it is mixed with the substrate and the enzymatic reaction takes place. Therefore, even in the case of a continuous enzymatic reaction in which the substrate is continuously fed into the container from the substrate solution inlet, most of the immobilized enzyme remains in the container and thus substantially contributes to the reaction mixture, i.e., the reaction mixture. Only product and unreacted substrate flow out of the reaction mixture outlet. Incidentally, even if some of the immobilized enzyme is not attracted to the magnet, this immobilized enzyme is separated from the reaction mixture by the separation membrane at the reaction mixture outlet. Therefore, according to the device of the present invention, even when enzymatic reactions are performed continuously, the immobilized enzyme is stirred and dispersed in the container under the attraction of the magnet, and is mixed with the substrate and reacts with the substrate. Because I will do it,
The reaction mixture remains substantially within the container as it flows out of the container. In this way, the separation load on the separation membrane is significantly reduced and clogging of the membrane is less likely to occur, so that the enzymatic reaction can be carried out continuously and stably over a long period of time.

Furthermore, if the rotating shaft is stopped, most of the immobilized enzyme is attracted to the magnet, so it is very easy to separate and recover the immobilized enzyme after the reaction is completed.
If necessary, part or all of the immobilized enzyme can be replaced together with the magnet.

FIG. 2 shows another embodiment of the enzyme reaction apparatus according to the present invention, in which a rotating shaft 5 is rotatably supported in a container 3, and a magnet 4 is attached to a stirring blade 6 of this rotating shaft. On the other hand, annular baffle plates 8 are attached to the inner wall surface of the container in multiple stages as necessary.

Therefore, in this device, when performing continuous enzyme reactions, the immobilized enzyme is dispersed and mixed by stirring by the stirring impeller and performs the enzyme reaction with the substrate, while most of the enzyme is absorbed by the magnet. stays in the container, so the separation load on the separation membrane is significantly reduced, similar to the above. Also, as described above, most of the immobilized enzyme is attracted and adhered to the magnet when the rotating shaft is stopped.

As described above, according to the enzyme reaction apparatus of the present invention, although the immobilized enzyme is mixed with the substrate solution under stirring by the rotary blade and an enzyme reaction is performed, most of the immobilized enzyme is kept inside the container by the magnet. Because it is retained, virtually only the reaction mixture can be obtained from the outlet, and even if some of the immobilized enzyme requires membrane separation, the separation load on the membrane is significantly reduced. clogging is unlikely to occur, and thus, according to the apparatus of the present invention, it is possible to carry out stable and continuous enzymatic reactions. Furthermore, if the rotation of the stirring blade is stopped, most of the immobilized enzyme will adhere to the magnet, making it easy to collect.
The immobilized enzyme can be easily replaced along with the magnet.

[Brief explanation of the drawing]

FIG. 1 is a sectional view showing one embodiment of the enzyme reaction apparatus according to the present invention, and FIG. 2 is a sectional view showing another embodiment of the enzyme reaction apparatus according to the invention. 1... Substrate solution inlet, 2... Reaction mixture outlet, 3...
Container, 4... Magnet, 5... Rotating shaft, 6... Stirring blade, 7
... Separation membrane, 8... Baffle plate.

Claims (1)

[Claims for Utility Model Registration] (a) A cylindrical container equipped with a substrate solution inlet and a reaction mixture outlet, and a separation membrane at the substrate solution inlet and reaction mixture outlet; (b) An axis of the cylindrical container. (c) a stirring blade that can rotate together with the rotary blade; (d) a magnet attached to the wall of the container or the stirring blade; The solution is supplied to the container, and while being stirred by a stirring blade in the container, particles having a diameter of 0.03 to 0.03 containing magnetic powder and immobilized enzymes are added.
1. An enzyme reaction device characterized in that the reaction mixture is stirred with water-dispersed polymer particles of 2μ, reacted with a substrate, and the generated reaction mixture flows out from the reaction mixture outlet.