JPS6371186A - Cultivation method - Google Patents

Abstract

PURPOSE:To efficiently cultivate a cultivation object assuming a specific form, by introducing the cultivation object into hollow pipes consisting of a membrane, impermeable to the cultivation object and permeable to a solute in a culture medium, hermetically sealing the above-mentioned pipes and entrapping the cultivation object. CONSTITUTION:A process system is kept in a germ-free state by using a thermal sterilization method with formaldehyde and steam and spawns are passed from a spawn tank through hollow fibers 5. A fresh culture medium is passed from a culture medium tank through the shell side of a hollow fiber module 1 to carry out cultivation to a given concentration. Fixed bolts 15 and head parts at both ends are aseptically removed from the module 1 and mounted in an inoculating pot 8. The hollow fibers 5 with a fixing plate 13 are passed from above through a fiber guide 6 and pushed down. A heating wire 7 as a heating source is moved in a direction crossing the hollow fibers 5 to cut and thermally fuse the hollow fibers 5 to a given length. The resultant entrapped material is then dropped into the inoculating pot 8 containing a previously introduced culture medium to keep microorganisms or cells in the entrapped body from inactivation.

Description

[Detailed Description of the Invention] [Industrial Application Field] The present invention relates to a culturing method, and in particular, the present invention relates to a method for culturing filamentous fungi, actinomycetes, plant tissues, etc., which, as a result of the progress of culturing, develop unique shapes such as fibrous or string-like shapes. The present invention relates to a culture method suitable for culturing a culture target that takes .

[Conventional technology]

Conventionally, culturing of filamentous fungi, actinomycetes, plant tissue, etc., has been carried out by aeration and mechanical stirring, or by aeration and agitation by aeration, as in the case of temporary microbial culture.

However, as the culture progresses, this type of culture target takes on a unique shape such as fibrous or string-like shapes, so there is a delay in response due to dirt on each sensor, and the power required for stirring due to the apparent increase in viscosity in the liquid. In addition, there are hardware problems such as an increase in the number of cells, as well as process problems such as shearing of the culture target due to mechanical stirring and an accompanying decrease in activity.

Therefore, in order to solve such problems, for example,
A technique has been proposed, as described in Japanese Patent No. 117085: - A technique is proposed in which a porous substance is made to exist in a solution as a breeding place for microorganisms.

[Problem that the invention seeks to solve]

In the above-mentioned conventional technology, the culture target is limited by the pore size of the porous material, it is difficult to increase the concentration of bacterial cells in the culture solution, and the bacterial cells are expelled from the porous material and float in the culture solution. There are problems such as the inability to prevent
There is still a problem that highly efficient culture of the above-mentioned culture targets cannot be performed.

An object of the present invention is to provide a culture method that can perform highly efficient culture of a culture target that takes a unique shape such as a fibrous or string shape as the culture progresses.

[Means for solving problems]

The above purpose is to provide a culture method including a step of introducing a culture target into a hollow tube made of a membrane through which the culture target cannot pass through and a solute in the medium can pass through, and sealing the end of the hollow tube. This is achieved by providing a method that includes the step of enclosing the culture target.

[For production]

A culture target is introduced into a hollow tube made of a membrane that is impermeable to the culture target and permeable to solutes in the medium.

Thereafter, the end of the hollow tube is sealed to enclose the culture target. As a result, even if the culture target takes on a unique shape such as a fibrous or string-like shape as the culture progresses, it can be treated and cultured in the same way as a culture target with a normal shape.

〔Example〕

Hereinafter, one embodiment of the present invention will be described in detail with reference to FIGS. 1 to 3, but the present invention is not limited thereto in any way.

Fig. 1 shows an outline of one embodiment of the pre-culture stage according to the present invention, Fig. 2 shows the transition operation from pre-culture to main culture, and Fig. 3 shows the hollow fiber module used in this example. The outline of each structure is shown below.

This device is mainly used for the Hollow 1 Eve module! ,
Culture medium tank 29 seeds bacterial membrane 3. It consists of an inoculation bot 8, and after pre-cultivating the desired microorganisms or cells in the hollow fiber 5 according to the flow shown in Fig. 1, the hollow fiber 5 is grown to an appropriate length by the operation shown in Fig. 2. The microorganisms or cells are then cut into pieces and held in an inoculation pot 8 as a microorganism or cell enclosing body. The hollow fiber 5 is formed of a membrane, such as a cellulose membrane, which is impermeable to the target microorganism or cell and permeable to solutes in the culture medium.

An outline of the operation according to this embodiment will be explained below.

At the time of pre-culturing, culturing is performed according to the flow shown in FIG. That is, first, formaldehyde (HCHO) is passed through the hollow fiber module 1 via the seed tank 3, and by circulating this, the inside of the module 1 is sterilized, and further, the inside of the module 1 is sterilized with process steam other than the module. Heat sterilize to make all process systems sterile. Next, the seed culture (SEED) is passed from the seed culture tank 3 into the hollow fiber 5. When the inside of the fiber 5 is filled with liquid, the valve on the outlet side is closed, and the piping and the inside of the seed tank 3 are washed with a buffer so that all the seed bacteria are retained in the hollow fiber 5. In this state, medium tank 2
Then, a fresh medium is passed through the shell side of the hollow fiber module 1, and the microorganisms or cells inside the fiber 5 are
A medium is supplied and the microorganisms or cells are cultured until they reach a predetermined concentration.

After the preculture is completed, the precultured microorganisms or cells are incorporated by the operation shown in FIG. Namely.

The fixing bolts 15 and the head portions at both ends are removed from the hollow fiber module l having the structure shown in FIG. 3 in a sterile atmosphere, and the module is attached to the inoculation pot 8 as shown in FIG. 2. From the top, hollow fiber 5
is pushed down together with the fiber fixing plate 13 through the fiber guide 6, and a chrome wire, platinum wire, etc.) is moved in a direction across the hollow fiber 5 to the heating wire 7 shown in FIG. Then, the obtained enclosing body is dropped into the inoculation pot 8. The culture medium or buffer of the preparation method is placed in advance in the inoculation pot 8, and the microorganisms or cells in the obtained inclusion body are maintained so as not to be inactivated.

In this example, the target microorganism or cell is enclosed in a hollow fiber, and the enclosing body is cultured as a unit, so that it can be treated in the same manner as other culture objects that take normal forms. Highly efficient culture can be performed even with ordinary culture equipment.

Furthermore, according to this example, microorganisms or cells can be precultured inside the membrane while being fed with fresh medium, and then main culture can be carried out under the same conditions, thereby reducing the activity of microorganisms or cells. (The culture can be continued.

In addition to the above-mentioned embodiments, the fiber may be introduced into the hollow fiber to be cultured after pre-culture and then sealed as described above to obtain comprehensiveness.

〔Effect of the invention〕

According to the present invention, even if a culture target takes on a unique shape such as a fibrous or string-like shape as the culture progresses, it can be treated in the same way as other culture targets that take a normal shape. Since it can be cultured, it has the effect of being able to perform highly efficient culture.

[Brief explanation of the drawing]

FIG. 1 is a flow diagram of a preculture device according to an embodiment of the present invention;
FIG. 2 is an external view of the enclosing device from the hollow fiber module of FIG. 1, and FIG. 3 is a vertical cross-sectional view of the hollow fiber module of FIG. 1. 1...Hollow fiber module, 2...
...Culture tl[,3...--Seed lli tank,5...
...Hollow fiber, ′; f2 diagram

Claims (1)

[Scope of Claims] 1. A step of introducing a culture target into a hollow tube made of a membrane through which the culture target cannot permeate and a solute in the medium can permeate, and sealing the end of the hollow tube. A culture method comprising the step of enclosing the culture target. 2. The culture method according to claim 1, wherein the end of the hollow tube is sealed by heat welding.