JPH07177A - Production device for macromolecular compound bearing n-acetylglucosamine residues - Google Patents

Abstract

PURPOSE:To provide the subject device which can recover macromolecular compound having N-acetylglucosamine residues efficiently in high productivity. CONSTITUTION:This production device has a horizontal cylinder vessel equipped with culture medium inlet, air inlet and culture mixture outlet is provided with a shaft so that it is connected to the external driving unit on its one end and the other contacts with the inner wall of the vessel on the center of the vessel in the longitudinal direction. The shaft is equipped with a plurality of perforated disks at a certain interval and one aeration unit is set to the bottom end of the shaft and another aerator is set right over the stirrer. In the upright cylinder vessel, two perforated cylindrical supports different in their diameter and mesh size are concentrically placed in the vessel so that their bottom end comes to 10 to 50mm above the vessel bottom, and an aeration pipe is set between the stirrer and the bottom end of the support and the pipe is connected through the joining pipe to the external pump.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an apparatus for producing a polymer compound having an N-acetylglucosamine residue, and particularly to a target N-acetylglucosamine residue by culturing a microorganism capable of producing the polymer compound. The present invention relates to an apparatus for efficiently producing a polymer compound having a group.

[0002]

BACKGROUND OF THE INVENTION In the present invention, the polymer compound having an N-acetylglucosamine residue means a compound produced by utilizing a microorganism. Cellulose is a polysaccharide in which only glucose is a constituent saccharide and is bound by β-1,4 bonds, but the present applicant has previously included an N-acetylglucosamine residue in the β-1,4-glucan chain of cellulose. A new polymer compound has been developed (JP-A-4-268301).
The polymer compound having an acetylglucosamine residue is
It refers to a novel polymer compound described in the publication. The production of a polymer compound having an N-acetylglucosamine residue utilizing a microorganism is usually performed by static culture. The reason for this is that it can be produced with a simple culture device, does not require agitation power, has low energy cost, and has good product quality. However, on the other hand, the production of a polymer compound having an N-acetylglucosamine residue by static culture has a drawback that the production rate is slow and it is not suitable for large-scale production. Then, this invention aims at provision of the apparatus used for manufacture of the high molecular compound which has an N-acetylglucosamine residue by aeration stirring culture.

The Applicant has conducted extensive studies to efficiently produce microbial cellulose similar to the novel polymer compound by static culture, and has formed a hydrophobic and oxygen-permeable membrane at the bottom of the culture solution. By arranging it, it was found that microbial cellulose can be produced not only on the surface of the culture solution but also on the surface of the culture solution to the same extent as that of the surface, and a patent application was filed (JP-A-5-123182). During the course of this research, microbial cellulose produced in the liquid, especially at the bottom of the culture, was attached to the membrane installed on the bottom of the device, which was attached to the membrane simply because of the high local oxygen concentration. However, the present invention was found to depend on the material of the membrane, and further, this finding was found to be applicable to a polymer compound having an N-acetylglucosamine residue, and the present invention was reached.

[0004]

The present invention relates to an apparatus for producing a polymer compound having the following N-acetylglucosamine residue. A horizontal cylindrical container having a medium inlet, a vent and a culture solution outlet, and a central part of the container so that one end of the rotary shaft is connected to a driving means outside the container and the other end is in contact with the inner wall of the container. A device for producing a polymer compound having an N-acetylglucosamine residue, characterized in that a plurality of perforated disk-shaped supports are attached to the rotary shaft in the longitudinal direction at fixed intervals.

In a vertical cylindrical container having a medium inlet and a culture medium outlet, a rotary shaft, one end of which is connected to a driving means outside the container, is inserted into the center of the container from the longitudinal direction, and the rotary shaft is rotated. An apparatus for producing a polymer compound having an N-acetylglucosamine residue, characterized in that ventilation means is provided at the lower end of the shaft, and a plurality of perforated disk-shaped supports are attached to the rotary shaft at regular intervals. .

[0006] In a vertical cylindrical container equipped with a medium inlet and a culture solution outlet and equipped with stirring means at the bottom,
A rotary shaft connected to a driving means outside the container is inserted into the center of the container from the longitudinal direction, and a plurality of perforated disk-shaped supports are attached to the rotary shaft at regular intervals, and the rotary shaft of the stirring means is directly connected to the rotary shaft. An apparatus for producing a polymer compound having an N-acetylglucosamine residue, characterized in that a ventilation means is provided on the upper part.

[0007] In a vertical cylindrical container equipped with a medium inlet and a culture solution outlet, and equipped with a stirring means at the bottom,
Two kinds of perforated hollow cylindrical supports having different diameters and mesh sizes are provided in the container, and the lower end is 10 to 50 m above the container bottom.
It is installed concentrically so as to be located at m, and a ventilation pipe is provided between the stirring means and the lower end of the support, and the pipe is connected to a pump outside the container via a connection pipe. For producing a polymer compound having an N-acetylglucosamine residue.

The size, material, etc. of the device of the present invention may be appropriately determined in consideration of the purpose of use, etc., but the perforated disk-shaped support and the perforated hollow cylindrical support are wire mesh-shaped and And a polymer compound having a produced N-acetylglucosamine residue is attached and retained. Therefore, the material of the support is required to have such ability. Specifically, in addition to metals such as stainless steel and titanium, Teflon, silicon, polyethylene, polypropylene, polyvinyl alcohol, polyacrylonitrile, polyimide, Polymers such as polystyrene are suitable, and these are used alone or in combination of two or more kinds. Also, the finer the mesh size, the better, but usually 3
-20, preferably 5-18 are suitable. When the support is thin and strength is required to be strengthened, the outer peripheral surface of the disk can be covered with a hard material such as stainless steel, titanium, aluminum, or a polymer in a ring shape.

Next, the apparatus for producing a polymer compound having an N-acetylglucosamine residue of the present invention will be described with reference to the drawings. As shown in FIG. 1, the above-mentioned device is mainly composed of a horizontal cylindrical container 1, which is provided with a medium inlet 2, a vent 3 and a culture medium outlet 4. Further, a rotary shaft 5 having one end connected to a driving means 6 outside the container is provided in the center of the container 1 so that the other end is in contact with the inner wall of the container, and the rotary shaft 5 has a wire mesh. A plurality of disk-shaped support bodies 7 are formed at regular intervals, usually 5 to 5.
10 are attached. Parts such as a drive clutch and a bearing are used for the rotary shaft 5. If necessary, a sensor inlet such as a pH composite electrode or an oxygen electrode can be provided. FIG. 2 shows the details of the device. The temperature of the liquid can be controlled by placing a jacket at the bottom of the container and circulating hot water from the constant temperature bath.

When a high molecular compound having an N-acetylglucosamine residue is produced by this apparatus, a medium is injected through the medium injection port 2 to inoculate the high polymer producing bacterium. In addition, it is desirable that the culture medium be soaked that the liquid surface is at the same level as the rotation axis. When sterilized air is introduced into the gas phase through the ventilation port 3 and the rotating shaft 5 is rotated by the driving means 6, the disc-shaped support 7 as a rotor also rotates at the same time, and the medium is stirred. Since the liquid surface of the medium is at the same level as the rotation axis, the upper part of the disk-shaped support 7 is normally exposed to the air, and is immersed in the medium by rotating it, while being immersed in the medium. The exposed part is exposed in the air by rotation. Therefore, oxygen is supplied not only directly to the microorganisms attached to the support in the gas phase, but also in the medium, so that oxygen is supplied very efficiently.

As shown in FIG. 3, the above-mentioned apparatus is mainly composed of a vertical cylindrical container 15, and although not shown, the container has a medium injection port, a ventilation port, and a culture solution withdrawal unit. There is a mouth etc. A rotary shaft 16 having one end connected to a drive means outside the container is inserted in the center of the container from the longitudinal direction, a ventilation means 17 is provided at the lower end of the rotary shaft, and a perforated disc-shaped support is provided on the rotary shaft. A plurality of bodies 18 are attached at regular intervals. The ventilation means 17 is like an air sparger, and may be of a type that takes in air or the like from the hollow portion of the rotary shaft 16 and supplies it into the apparatus, or via a pipe separate from the rotary shaft. You may make it take in air etc. from the outside. Also, the number of supports may be the same as in the case of the above device.

As shown in FIG. 4, the above-mentioned apparatus is mainly composed of a vertical cylindrical container 19, and although not shown, the container has a medium inlet, a vent and a culture medium take-out port. A mouth and the like are provided, and a stirring means 20 (a magnetic stirrer is used in the case shown) is provided at the bottom. A rotary shaft 21 having one end connected to a driving means outside the container
Is inserted into the center of the container from the longitudinal direction, and a plurality of perforated disk-shaped supports 22 are provided on the rotation shaft at regular intervals (usually 3 to
(7 pieces) are attached. A ventilation means 23 is attached directly above the stirring means, and is connected to an external air supply means via a suitable pipe to introduce air or the like into the apparatus.

As shown in FIG. 5, the above-mentioned device is mainly composed of a vertical cylindrical container 24, and although not shown, the container has a medium inlet, a vent and a culture medium withdrawal port. A mouth and the like are provided, and a stirring means 25 (a magnetic stirrer is used in the case shown) is provided at the bottom. Two kinds of perforated hollow cylindrical supports 26 and 27 having different diameters and mesh sizes are provided so that the lower end is above the container bottom 10-5.
It is installed concentrically so that it is located at 0 mm. A ventilation pipe 28 is provided between the stirring means and the lower end of the support.
Is provided, the pipe is connected to a pump outside the container through a connecting pipe, and air or the like is introduced into the apparatus.

The production of microbial cellulose, the production of other insoluble microorganism-produced polymers, the production of vinegar, etc. using the apparatus described in any one of (1) to (3) of the present invention can be basically carried out in the same manner as described for the above apparatus. it can. N produced by culturing a polymer compound-producing bacterium having an N-acetylglucosamine residue using the device of the present invention
-The polymer compound having an acetylglucosamine residue is
It is attached to the disk-shaped support (device of ~) or the hollow cylindrical support (device of it), and is hardly present in the culture solution or on the surface of the culture solution. Therefore, after completion of the culture, the culture containing the polymer compound is peeled from the support, and the polymer compound having an N-acetylglucosamine residue is recovered by an appropriate means. As an example, the culture is dipped in a sodium dodecyl sulfate solution and boiled to remove proteins derived from bacterial cell components and medium components, and then immersed in an aqueous solution of an alkaline substance such as caustic soda, and further, such as acetic acid. There is a method of immersing in an acidic solution, neutralizing, and then washing with water.

[0015]

INDUSTRIAL APPLICABILITY Since the apparatus for producing a polymer compound having an N-acetylglucosamine residue of the present invention has a wire-mesh-like support installed in the apparatus in the form of a disc or a hollow cylinder, The productivity of the polymer compound having an N-acetylglucosamine residue can be increased as compared with the case of culturing the polymer compound-producing bacterium without using these. Further, the polymer compound having an N-acetylglucosamine residue produced by using this apparatus is efficiently attached to and retained on the wire mesh support.

[Brief description of drawings]

FIG. 1 is an explanatory view showing one embodiment of the device of the present invention.

FIG. 2 is an explanatory diagram showing details of the apparatus of FIG.

FIG. 3 is an explanatory view showing one embodiment of the device of the present invention.

FIG. 4 is an explanatory view showing one embodiment of the device of the present invention.

FIG. 5 is an explanatory view showing one embodiment of the device of the present invention.

[Explanation of symbols]

 1: Horizontal cylindrical container 2: Medium injection port 3: Vent port 4: Culture solution extraction port 5: Rotating shaft 6: Drive device 7: Perforated disk-shaped support 8: Drive clutch 9: Rotor bearing 10: Fixed Disc flange 11: Sensor inlet 12: Speed control pack 13: Device fixing plate 14: Substrate 15: Vertical cylindrical container 16: Rotating shaft 17: Venting means 18: Perforated disc support 19: Vertical cylinder Container 20: stirring means 21: rotating shaft 22: perforated disk-shaped support 23: ventilation means 24: vertical cylindrical container 25: stirring means 26: perforated hollow cylindrical support 27: perforated hollow cylindrical Support 28: ventilation means

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Masahiro Fukaya 28, No. 1, Muraoka, Higashiura-machi, Chita-gun, Aichi Prefecture (72) Inventor Yuko Kanegae 14, 2-27, Ganjuku-cho, Handa, Aichi 102 102 Parkside Hills No. (72) Inventor, Hitoshi Okumura, 14-5-66, Iwanamehigashi-cho, Handa-shi, Aichi Prefecture (72) Inventor, Yoshiya 132, Furuwato, Kochino-cho, Konan-shi, Aichi Prefecture

Claims (5)

[Claims] 1. A horizontal cylindrical container having a medium inlet, a vent and a culture solution outlet so that one end of the rotary shaft is connected to a driving means outside the container and the other end is in contact with the inner wall of the container. A polymer compound having an N-acetylglucosamine residue, which is provided in the central portion of the container in the longitudinal direction, and a plurality of perforated disk-shaped supports are attached to the rotation shaft at regular intervals. Manufacturing equipment. 2. The device according to claim 1, which has a sensor inlet for a pH composite electrode, an oxygen electrode, or the like. 3. A vertical cylindrical container provided with a medium inlet and a culture solution outlet, and a rotary shaft, one end of which is connected to a driving means outside the container, is longitudinally inserted into the center of the container, Ventilation means is provided at the lower end of the rotary shaft, and a plurality of perforated disk-shaped supports are attached to the rotary shaft at regular intervals, to produce a polymer compound having an N-acetylglucosamine residue. Equipment. 4. A vertical cylindrical container having a medium inlet and a culture medium outlet and having a stirring means at the bottom,
A rotary shaft connected to a driving means outside the container is inserted into the center of the container from the longitudinal direction, and a plurality of perforated disk-shaped supports are attached to the rotary shaft at regular intervals, and the rotary shaft of the stirring means is directly connected to the rotary shaft. An apparatus for producing a polymer compound having an N-acetylglucosamine residue, characterized in that a ventilation means is provided on the upper part. 5. A vertical cylindrical container provided with a medium inlet and a culture medium outlet and having a stirring means at the bottom,
Two kinds of perforated hollow cylindrical supports having different diameters and mesh sizes are provided in the container, and the lower end is 10 to 50 m above the container bottom.
It is installed concentrically so as to be located at m, and a ventilation pipe is provided between the stirring means and the lower end of the support, and the pipe is connected to a pump outside the container via a connection pipe. For producing a polymer compound having an N-acetylglucosamine residue.