JPH03219866A - Culturing method and device therefor - Google Patents

Abstract

PURPOSE:To make it possible to select an optimum operation state in series and in period of culture and make it possible to prevent the increase of microbial cell adhering to wall face by making a draft tube opened in the upper and lower ends of a vertical culture vessel changeably either into a state of cylinder with no opening on the side face or a state having a number of openings. CONSTITUTION:A draft tube 1 of foam tower type vertical culture tank 2 is provided in such a structure changeable either into a state of cylinder as shown in Fig. I or a state having opening in vertical direction as shown in Fig. II. When it is desired to keep the state of a bacterial or cellular suspension in the earlier stage of culture preferable while using this culture tank, the draft tube 1 is changed into the state expressed by Fig. II and bacteria or cells are cultured by subjecting to an aerating cultivation while stirring by a stirring wing in a draft tube 1. In the middle state or latter stage of culture, the draft tube 1 is changed into the state expressed by Fig. I and the bacteria or cells are cultured while operating the draft tube 1 as a foaming tower so as to reduce the damage to the bacteria or cells.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to a method and apparatus for efficiently culturing microbial cells, cells, etc. in a culture solution.

[Conventional technology]

The mainstream of conventional culture equipment is to fill a tank with a culture solution containing bacteria, cells, etc., stir it with a stirrer, and blow in gas such as air from the bottom of the tank to circulate the culture solution and create air-liquid contact. This was a so-called aerated stirring method in which the reaction proceeded by performing the following steps.

However, depending on the type of culture, the strong shearing force caused by stirring may have a large effect on the bacterial bodies or cells. Furthermore, there is a need to reduce the installation area, and the application of vertical (tower-shaped) culture tanks to various culture systems is being considered.

The mainstream of base-type culture tanks is to install a draft tube inside the tank.
It was a bubble column method in which a tank was filled with a culture solution containing bacterial bodies, cells, etc., and gas such as air was blown from the bottom of the tank to circulate the culture solution and bring about gas-liquid contact.

Regarding the conventional bubble column method, Japanese Patent Application Laid-open No. 57-39771 discloses that the cross section of the inner cylinder of the aeration tank is divided into an appropriate number of parts to form divided cylinder parts extending in the vertical direction, and an air supply port is provided at the bottom of the divided cylinder part. A fluidized bed type biological treatment apparatus is disclosed.

Furthermore, Japanese Patent Application Laid-Open No. 63-251077 discloses a culture device that has an axial flow impeller installed inside a cylinder in a culture tank, and rotates this impeller to efficiently culture cells, etc. with a small amount of power. Disclosed.

Furthermore, in Japanese Patent Application Laid-open No. 1-150476, a porous draft tube with open upper and lower ends is provided in the container,
A bubble column reactor is disclosed in which gas is blown into the draft tube from below.

[Problem to be solved by the invention]

The conventional bubble column method described above has the following disadvantages.

fil Overall, since the flow is relatively gentle, it is often insufficient to efficiently suspend bacterial bodies, cells, etc. in the entire system at the initial stage of culture.

(2) Since the direction of fluid flow is almost constant, bacterial bodies or cells tend to adhere to the wall surface, resulting in a large number of growths adhering to the wall surface.

(3) Early stage of culture, middle stage of culture, late stage of culture, liquid composition,
When oxygen demand changes over time, the controllable range is narrow.

In addition, the above-mentioned Japanese Patent Application Laid-Open No. 57-39771, Japanese Patent Application Laid-Open No. 6
3251077 and Japanese Patent Application Laid-Open No. 1-160476, the characteristics of the present invention are described as follows: ``Depending on the culture state, the draft tube is a cylinder with no openings on the side, and the draft tube has many openings on the side. The technical idea of ``switching to a state with ``is not disclosed or suggested at all.

The present invention was made in view of the above points, and it has a draft tube that is cylindrical like the draft tube of a conventional bubble column type vertical (tower type) culture tank, and a gap is formed in the vertical direction. By having a structure that can be adjusted as appropriate depending on the condition, it is possible to appropriately select the optimal operating condition for the type of culture and culture period, and it is possible to prevent an increase in the number of bacterial cells adhering to the wall surface. It is an object of the present invention to provide a method and apparatus.

[Means to solve the problem]

In order to achieve the above object, the culture method of claim 1 includes:
As shown in FIGS. 1 to 6, a draft tube 1 with open upper and lower ends is installed in a vertical culture tank 2 so that a gap is created between the tank side wall, the tank bottom wall, and the tank lid, Fill the draft tube and culture tank with the culture solution 3 containing bacteria, cells, etc., stir the inside of the draft tube, and blow gas from below the draft tube to circulate the culture solution and create air-liquid contact. In the culture method to be carried out, depending on the culture state, the draft tube 1 is switched between a state (a) in which the draft tube 1 is a cylindrical body with no openings on the side surface and a state in which the draft tube 1 has a large number of openings 13 in the side surface (b). It is characterized by

The culture method according to claim 2 is the culture method according to claim 1, as shown in FIGS.
When the draft tube is switched to a state with a large number of openings 13, most of the bacterial cells during culture, floc-like bacterial cells, immobilized bacterial cells, or animal/plant cells are attached or filtered out. It is characterized by

As shown in FIGS. 1 to 6, the culture apparatus according to claim 3 is a vertical culture system in which a draft tube 1 whose upper and lower ends are open is filled with a culture solution 3 containing bacterial bodies, cells, etc. A gap is provided in the tank 2 between the tank side wall, the tank bottom wall, and the tank lid, and one or more stirring tubes 1!1 are installed in the draft tube 1.
11 and a gas blowing nozzle 12 connected below the draft tube, the culture apparatus is characterized by having a large number of openings 13 on the side surface of the draft tube 1, which can be opened and closed.

As shown in FIGS. 7 to 10, the culture device according to claim 4 is the culture device according to claim 3, in which bacterial cells and floc-like bacteria during culture are placed inside or outside the opening on the side of draft tube 1. The present invention is characterized by the provision of silk or a porous material 18 having a pore size and material to which most of the immobilized bacterial cells or animal/plant cells can be attached or filtered.

As shown in FIGS. 11 to 14, the culture device according to claim 5 has a draft tube 1 whose upper and lower ends are open, and which is used to store bacterial cells,
A vertical culture tank 2 filled with a culture solution 3 containing cells, etc. is provided so that a gap is created between the tank side wall, tank bottom wall, and tank lid, and one or more stirrers 11 are installed in the draft tube 1. In a culture apparatus in which a gas blowing nozzle 12 is connected to the lower part of the draft tube, the draft tube 1 is formed into a louvered cylinder by supporting a large number of longitudinal strips 20, and these strips 20 are pivoted. It is characterized in that it can be rotated around the support shaft 21 to perform open vA chants.

As shown in FIGS. 15 and 16, the culture apparatus of claim B includes a vertical culture tank 2 in which a draft tube 1 with open upper and lower ends is filled with a culture solution 3 containing bacterial bodies, cells, etc. One or more stirrers 11 are installed inside the draft tube 1, and a gas blowing nozzle 12 is installed below the draft tube.
In the culture device connected to the draft tube, the draft tube is formed from an inner cylinder 15 having a large number of vertical slits 14 and an outer cylinder 17 having a large number of vertical slits 16,
It is characterized in that the draft tube can be adjusted to open and close by rotating the outer cylinder or the inner cylinder.

[Effect]

Culture When it is desired to improve the suspended state of bacterial bodies, cells, etc. described above, the state is switched to state (b) and the draft tube is operated as an aeration stirring tank while being stirred by a stirring blade and cultured.

In the middle and late stages of culture, in order to reduce damage to bacterial bodies, cells, etc., the state is switched to state (a) and cultured by operating as a bubble column. Note that it can be appropriately selected whether the stirring blade in the draft tube should be stopped or stirring should be performed.

When the number of adhering and proliferating bacterial bodies, cells, etc. on the wall increases, the state of Li
Switch to state (b) to remove the adhesion, then switch to state (a).
) and continue culturing.

〔Example〕

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the drawings. However, unless there is a specific description, the shapes of the components described in this example, their relative positions, etc. are not intended to limit the scope of the present invention to these, but are merely illustrative examples. Only.

FIGS. 1 and 2 show an embodiment of the culture device of the present invention, and FIG. 1 is a longitudinal cross-sectional view showing a state (a) in which the draft tube is a cylinder with no opening on the side, for example, a cylinder. FIG. 2 is an explanatory longitudinal cross-sectional view showing a state in which the draft tube has a large number of openings on the side surface [(b). Also, the third
The figure is an explanatory cross-sectional view of the apparatus shown in FIG. 1, and FIG. 4 is an explanatory cross-sectional view of the apparatus shown in FIG.

1 is a draft tube whose upper and lower ends are open. 2
is a vertical culture tank, which is filled with a culture solution 3 containing bacterial bodies, cells, etc. The draft tube 1 is inserted into the culture tank 2, and the tank side wall 4
, are fixed so that a gap 7.8.10 is created between the tank bottom wall 5 and the tank M6. And draft tube 1
One or more stirrers 11 are provided inside the draft tube 1, and a gas blowing nozzle 12 is connected below the draft tube 1.

The above is the configuration of a conventional culture device, and in this culture device, a large number of openings 13 that can be opened and closed are provided on the side surface of the draft tube 1. Draft tube 1 is
For example, as shown in FIG. 3 and FIG.
3, or as shown in FIGS. 5 and 6, an inner cylinder 15 having a large number of vertical slits 14 and an outer cylinder 1 having a large number of vertical slits 16.
7, and the draft tube 1 can be opened and closed by rotating the outer cylinder 17 or the inner cylinder 15.

1a is a draft tube. Note that the mechanism of the louver-like structure will be explained in detail later.

FIGS. 7 to 10 show other embodiments of the culture apparatus of the present invention. In the culture device of this example, most of the bacterial cells, floc-like bacterial cells, immobilized bacterial cells, or animal/plant cells during culture are attached or separated inside the opening on the side of the draft tube 1 or 1a. A net or porous material 18 of a suitable pore size and material is provided. Other configurations and operations are the same as those in FIGS. 1 and 2.

11 to 14 show an example of an opening/closing mechanism for a louver-like structure. The draft tube 1 is formed into a louvered cylinder by pivoting a number of longitudinal strips 20, and is opened and closed by rotating these strips 20 around a pivot shaft 21.
j1 section. That is, a plurality of vertical operating frames 22 are provided around the draft tube 1 made of a louver-like structure, and a ring 23 is provided at the lower end of these operating frames 22 and below the draft tube 1. A plurality of push fittings 24 (same number as the number of louvers) are fixed to the upper part of the draft tube 23. Also, a ring 25 is arranged above the draft tube 1, and this ring 25 is fixed to the operating frame 22. A plurality of push fittings 26 (same number as the number of louvers) facing downward are fixed to this ring 25. The upper pivot shaft fixing ring 27 and the lower pivot shaft fixing ring 28 allow the pivot shaft to partially pass through the louver. 30 is fixed so that the louver can be opened and closed freely by the operation of the pusher fittings 26.The upper and lower pivot shafts 30 of the louvers are connected to the louver fixing fittings 31 to prevent a portion of the louvers from freely moving up and down.
Set and fix. In addition, 32 is a tank lid.

As shown in FIG. 1111iffl and FIG. 13, when the operating frame 22 is pulled up by human power or driving force such as a motor, the louver is pushed from inside by the pusher 24 at the bottom of the operating frame 22, closing the louver and making it into a cylindrical shape.

Further, as shown in FIGS. 12 and 14, the operating frame 22
When the louver is pulled down, the louver is pushed from the outside by a pusher fitting 26 on the upper part of the operating frame 22, and the louver is opened.

The 15th and 16th sections show a structure in which a mesh or porous material 18 is provided inside the draft tube 1 in the culture apparatus shown in FIGS. 11 to 14.

In this case, as shown in FIGS. 15 and 16, it is desirable to join one end of the louver to a part of the mesh or porous material 18, where 33 is a joint.

Next, using the culture apparatus shown in FIGS. 11 to 14,
The results of the culture test will be explained.

Medium composition: tannic acid: 2.0 wt%, (NH,
)H2PO4: ], 4 iyt%, KHzPOn:
o, 2 wt%, MgSO4' 7HzO: 0.
Aspergillus oryzae strain; Aspergillus oryzae
A vertical culture tank with an internal volume of 2000 cc was filled with 1,650 cc of a liquid inoculated with 25 μ/l (as wheat koji) of M. yzae), and air was blown at an aeration rate of 3.3 β/min. A stirrer was installed inside the draft tube.

First, the draft tube is made into a cylindrical state with no openings on the side (state (a)), air is blown into it, the agitator is stopped, and it is used as a bubble column.The culture time (h) and the dry bacterial weight ( g/L) and the amount of tannase produced (CU)
/~) The relationship was as shown in FIG. In addition, U/a/ is 1 Jnit/d (titer).

Next, change the draft tube to a state with many openings on the side (state (b)), blow in air, and rotate the stirrer at 380 rpm to use it as an aeration stirring tank. Dry bacterial weight it (g
/L) and the amount of tannase (U/d) as a product, as shown in FIG.

From FIG. 17, it can be seen that the rise time is shorter when used as an aeration stirring tank than when used as a bubble column.

〔Effect of the invention〕

Since the present invention is configured as described above, it achieves the following effects.

(1) While making the most of the features of a vertical (base type) culture tank, it is possible to add the advantages of an aeration agitation system.

(2) Optimal conditions can be appropriately selected depending on the type of culture and culture time.

(3) It is possible to prevent an increase in bacterial cells adhering to the wall surface.

(4) In the case of claim 2.4, bacterial bodies, cells, etc. are filtered and concentrated on the surface of the net or porous material.

(5) In the case of claim 5, the shape of the draft tube can be changed from the outside during the culture.

[Brief explanation of drawings]

Figures 1 and 2 show an embodiment of the culture device of the present invention. Figure 1 is a vertical cross-sectional view showing the side opening of the draft tube in a closed state, and Figure 2 is a side view of the draft tube. 3 is an explanatory cross-sectional view of the device shown in FIG. 1, the fourth leap is an explanatory cross-sectional view of the device shown in FIG. 2, and FIGS. The figure is a cross-sectional explanatory diagram showing another embodiment of the culture device of the present invention, and Figures 111EI and 12 show an example of an opening/closing mechanism for the side opening of the draft tube in the culture device of the present invention. The figure is an explanatory elevational view showing the side opening of the draft tube in the closed state, FIG. 12 is an explanatory elevational view showing the opening on the side of the draft tube in the open state, and FIG.
' line cross-sectional explanatory diagram, Fig. 14 is B-B in Fig. 12.
' Line cross-sectional explanatory diagrams, Figures 15 and 16 are illustrations of the present invention.
FIG. 17, an explanatory diagram showing still another embodiment of the apparatus, shows an example of culture test results, and is a graph showing the relationship between culture time, dry bacterial weight, and tannase amount. 1.1a... Draft tube, 2... Vertical culture tank, 3... Culture solution, 4... Tank side wall, 5... Tank bottom wall,
6... Tank lid, 7.8.10... Gap, 11... Stirrer, 12... Gas blowing nozzle, 13... Opening,
14.16...Slit, 15...Inner cylinder, 17...
・Outer cylinder, 18... Net or porous material, 20... Strip, 2
1... Pivot shaft, 22... Operating frame, 23.25...
Ring, 24. 26... Push metal fitting, 27... Upper pivot shaft fixing ring, 28... Lower pivot shaft fixing ring, 3
G... Pivot shaft, 31... Louver fixing bracket, 32...
・Tank lid, 33...Joint part ζ;;

Claims (1)

[Claims] 1. A draft tube (1) with open upper and lower ends is provided in a vertical culture tank (2) so that a gap is created between the tank side wall, the tank bottom wall, and the tank lid. Fill the tube and culture tank with a culture solution (3) containing bacterial bodies, cells, etc., stir the inside of the draft tube, and blow gas from below the draft tube to circulate the culture solution and make air-liquid contact. In the culture method, depending on the culture state, there is a state (a) in which the draft tube (1) is a cylinder with no openings on the side, and a state in which the draft tube (1) has a large number of openings (13) on the side. (b) A culture method characterized by switching to. 2. A net or porous material (18) is provided inside or outside of the many openings on the side of the draft tube (1), and when the draft tube is switched to a state with many openings (13), 2. The culturing method according to claim 1, wherein most of the bacterial cells, floc-like bacterial cells, immobilized bacterial cells, or animal/plant cells are attached or separated. 3 Place a draft tube (1) with open upper and lower ends into a vertical culture tank (2) filled with a culture medium (3) containing bacteria, cells, etc., and place it on the tank side wall, tank bottom wall, and tank lid. In a culture device in which a draft tube (1) is provided with a gap between the two, one or more stirrers (11) are provided in the draft tube, and a gas blowing nozzle (12) is connected below the draft tube.
A culture device characterized in that a draft tube (1) is provided with a number of openings (13) on the side thereof that can be opened and closed. 4 Pore size and material that allow most of the cultured bacteria, floc-like bacteria, immobilized bacteria, or animal/plant cells to adhere to or be filtered out on the inside or outside of the opening on the side of the draft tube (1). 4. The culturing device according to claim 3, further comprising a net or porous material (18). 5 Place a draft tube (1) with open upper and lower ends into a vertical culture tank (2) filled with a culture medium (3) containing bacteria, cells, etc., and place it between the tank side wall, tank bottom wall, and tank lid. In a culture device in which a draft tube (1) is provided with a gap between the two, one or more stirrers (11) are provided in the draft tube, and a gas blowing nozzle (12) is connected below the draft tube.
Draft tube (1) with a number of longitudinal strips (20)
are pivoted to form a louvered cylinder, and these strips (2
0) can be rotated around a pivot shaft (21) to adjust opening and closing. 6 Insert a draft tube (1) with open top and bottom ends into a vertical culture tank (2) filled with a culture medium (3) containing bacteria, cells, etc., along with the tank side wall, tank bottom wall, and tank lid. In a culture device in which a draft tube (1) is provided with a gap between the two, one or more stirrers (11) are provided in the draft tube, and a gas blowing nozzle (12) is connected below the draft tube.
Cut the draft tube into many longitudinal slits (14)
an inner cylinder (15) with a large number of longitudinal slits (15);
6) and an outer cylinder (17), and the draft tube can be opened and closed by rotating the outer cylinder or the inner cylinder.