KR20170013443A - Recovery System for bacterial cellulose culture medium and manufacture method for bacterial cellulose using the same - Google Patents

Abstract

The present invention relates to a device for collecting a bacterial cellulose culture medium, and to a method for producing bacterial cellulose by using the device. The device comprises: a plurality of culturing trays staked in multiple layers and culturing microorganisms; a hydraulic unit for making the culturing trays slantly inclined; a liquid collecting line connected with each of the culturing trays, and collecting a culture medium; a filter for removing impurities in the culture medium in which the liquid collecting line is integrated and connected as one; a culturing tank for storing and culturing the culture medium and connected with the filter; a vacuum pump for moving the culture medium and connected with the culturing tank; and a liquid collection adjusting device for controlling the collection and feeding of the culture medium. According to the present invention, the device collects the culture medium discarded after culturing is finished, and feeds a sterilized culture medium to the collected culture medium, so bacterial cellulose can be continuously cultured without having a pre-culturing step. Moreover, when bacterial cellulose is produced by using the device, a larger quantity of bacterial cellulose which is thicker can be produced within a period compared to a conventional process of producing bacterial cellulose. Also, fewer amount of culture media are discarded, so productivity can be improved.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a bacterial cellulose culture medium recovery apparatus and a bacterial cellulose production method using the same.

The present invention relates to a bacterial cellulose culture liquid recovery apparatus and a method for producing a bacterial cellulose using the same, and more particularly, to a bacterial cellulose culture recovery apparatus comprising a plurality of culture trays for stacking a plurality of layers and culturing microorganisms, a hydraulic jaw for inclining the culture tray, A liquid recovery line connected to each culture tray for recovering the culture liquid, a filter for removing impurities from the culture liquid in which the liquid recovery lines are integrally connected, a culture tank for storing and culturing the culture liquid connected to the filter, A vacuum pump for moving the culture liquid connected to the tank, and a liquid recovery regulating device for controlling recovery and injection of the culture liquid, and a method for producing bacterial cellulose using the recovering device will be.

Cellulose is the most abundant biomaterial resource in the natural world. In particular, the cellulose produced by the acetic acid bacteria is attracting much attention as a high value-added industrial new material as well as a food additive. In particular, unlike cellulose produced by plants, cellulose produced by bacteria is a high-purity cellulose that does not contain hemicellulose or lignin, and has excellent physicological properties, and much research has been conducted on the development of practical materials.

The microbial cellulose has a three-dimensional network structure of fibrils having a primary structure of 20 to 50 nm in? -1,4-glucan. Such microbial cellulose may be produced by a microorganism belonging to the genus of Acetobacter, Rhizobium or Agrobacterium, preferably produced by an microorganism belonging to the genus Acetobacter, It is produced by xylinum (Acetobacter xylinum).

The manufacturing process of the bacterial cellulosus used in the conventional mask pack is long in about 15 days until the bacterial cellulosus is produced. In addition, since the amount of the remaining cell culture liquid after the production of the bacterial cellulosus by the political culture is large, the installation cost of the wastewater treatment facility and the waste water treatment cost are incurred.

Korean Patent No. 10-1109034 relates to a mass production method of microbial cellulose and a manufacturing method of a mask pack using the same. In order to produce a mask pack using microbial cellulose, the cellulose produced in the culture medium for mass culture is collected, It is described that one cellulosic is molded again with a molding machine to produce a mask pack. Korean Patent No. 10-1371408 relates to a system and a method for manufacturing a cellulose mask, and a method for manufacturing a mask pack according to an automated facility is disclosed. That is, the patents relate to methods for producing microbial cellulose.

On the other hand, the present invention relates to a device for continuous culture of recovered bacterial cellulase from a remaining lung culture medium, which is not mentioned or known at all.

Accordingly, the present inventors have made intensive studies to overcome the problems of the prior art. As a result, the inventors have found that there are a plurality of culture trays for stacking multi-layers and culturing microorganisms, a hydraulic jaw for tilting the culture trays, A filter for removing impurities from the culture solution, a culture tank for storing and culturing the culture solution connected to the filter, a culture tank connected to the culture tank for connection to the culture tank, In the case of the bacterial cellulose culture liquid recovery device of the present invention comprising a vacuum pump for movement of the culture liquid and a liquid recovery control device for controlling the collection and injection of the culture liquid, the culture liquid discarded after the completion of the culture is recovered, The culture medium was then transferred to the culture medium It is possible to cultivate bacterial celluloses in the same manner as in the case of producing bacterial celluloses using the above apparatus and to produce thicker and larger amounts of bacterial cells It is possible to produce roulocytes, and the culture medium to be discarded can be reduced, resulting in an increase in productivity. Thus, the present invention has been completed.

KR 10-1109034 B KR 10-1371408 B

Accordingly, it is a main object of the present invention to provide a method for recovering a discarded culture liquid after cultivation and continuously culturing the bacterial cellulosus without injecting the sterilized medium into the recovered culture solution, Which is capable of producing a thicker and larger amount of bacterial celluloses within the same period of time as compared with the process of producing real cellulosic cells and capable of increasing the productivity due to a small amount of culture liquid to be discarded.

It is another object of the present invention to provide a continuous production method of bacterial cellulose using the bacterial cellulose culture liquid recovery apparatus.

According to one aspect of the present invention, there is provided a method for culturing a microorganism, comprising: a plurality of culture trays for stacking a plurality of layers to culture microorganisms;

A hydraulic jacket for tilting the culture tray;

A liquid recovery line connected to each of the culture trays to recover the culture liquid;

A filter for removing impurities from the culture fluid in which the liquid recovery lines are integrally combined and connected;

A culture tank for storing and culturing the culture fluid connected to the filter;

A vacuum pump for moving the culture fluid connected to the culture tank; And

And a liquid recovery control device for controlling the recovery and injection of the culture liquid.

The inventors of the present invention found that the number of viable cells of the culture solution, which was discontinued after the production of the bacterial cellulose by the conventional method without using the recovery device, was much earlier than that of the culture solution, and the culture solution thus discarded was aseptically recovered and reused As a result, the inventors of the present invention invented a bacterial cellulose culture liquid recovery apparatus.

In the present invention, the microorganism is characterized by producing cellulolose secreted outside the cell. Preferably, the microorganism is selected from the group consisting of Acetobacter sp., Rhizobium sp., Agrobacterium sp. , Pseudomonas sp., Alcaligenes sp., Azotobacter sp., Agrobacterium sp., Sarcina sp., And the like. The polysaccharide produced by the microorganisms is characterized by being cellulose.

Conventionally, since the above-mentioned microorganisms were cultured to produce celluloses in the upper part of the culture solution, and the resulting celluloses were recovered and the culture solution in the lower layer was discarded, the amount of culture liquid discarded was large. However, the inventors of the present invention have developed a device for recovering the culture solution after discontinuing the reaction, and recycled it.

The present invention is further characterized in that a plurality of culture trays, a liquid recovery device, and a cradle equipped with a moving wheel are provided at lower corners so as to move the filter.

In the present invention, the hydraulic device is connected to a lower portion of a cradle of the tray. The hydraulic apparatus is located on the opposite side of the liquid recovery line, and a plurality of culture trays are lifted by 2 to 3 cm in the direction of the liquid recovery line so that the culture liquid can be collected on one side.

In the present invention, the tray may have a liquid recovery line inlet formed on a side of the tray so that the liquid recovery line can be inserted therein, and the inlet is preferably formed on the opposite side of the hydraulic apparatus.

In the present invention, the plurality of trays may be composed of one or more trays, but the number of trays is preferably 10 to 60, but is not limited thereto.

In the present invention, the liquid recovery line may be any flexible material, preferably silicon. Since the liquid recovery line is inserted into the liquid recovery line insertion port to absorb the culture liquid, it is preferable that the liquid is well bent so as to absorb the culture liquid as much as possible.

In the present invention, the liquid recovery line is made of a material that can withstand steam, and is preferably made of silicon. Since the liquid recovery line of the present invention is sterilized by steam treatment so as not to be contaminated by other bacteria, it is preferably a material which is not damaged by steam treatment.

The present invention is characterized in that a valve is formed in each of the liquid recovery lines connected to the plurality of trays. When the amount of the plurality of trays to be used is small as compared with the liquid recovery line provided, the valve can lock the valve of the unused liquid recovery line to prevent unnecessary consumption.

In the present invention, the filter is formed so as to be able to remove impurities other than cells. Specifically, the material of the filter is a metal and the size of the hole is about 0.5 to 1.5 mesh.

In the present invention, the apparatus collects all of the culture liquid from the culture tray, inoculates the culture liquid into the sterilized culture medium, removes the existing plurality of culture trays, mounts a new plurality of trays, Is injected into the culture medium. By producing the bacterial cellulose in a new tray using the pulp culture solution, it is possible to produce more bacteriorcellulose of better quality within the same time period as the conventional production period of the bacterial cellulose, and the waste water treatment cost can be reduced.

According to another aspect of the present invention, the present invention provides a continuous production method of bacterial cellulose using the apparatus of the first aspect.

a) preparing a bacterial cellulose using a tray;

b) recovering the cultured liquid from the tray of step a) after the production of the bacterial cellulose by using the culture solution recovery device of claim 1;

c) re-dispensing and culturing the medium of step b) in a new tray; And

d) repeating steps b) and c) three to six times to produce a bacterial cellulose.

The pulmonary culture solution recovered by using the bacterial cellulose culture liquid recovering apparatus of the present invention can be further divided into a new tray and cultured to produce more bacterial cellulose during the production of the bacterial cellulose, The processing cost can be reduced.

In step (a), the strain producing bacterial celluloses is cultivated in a fermenter, and the strain is dispensed into a tray and cultured at 26 to 32 ° C for 4 to 5 days to prepare bacterial celluloses.

In the step (b), since only the collection line of the recovery device, which is steam-sterilized without opening the tray, is inserted into the tray, contamination by external bacteria is minimized and only the culture liquid is recovered.

The culture inoculated in step (c) is divided into 2 to 3 days by culturing for 4 to 5 days in a conventional manner without dividing the culture into a tray and terminating the culture.

The culture medium recovered in the step (b) is capable of shortening the culture period compared to the conventional culture period in the step (c) due to an increase in the number of bacterial cells producing bacterial celluloses as compared with the conventional culture medium inoculated.

In the present invention, the method further comprises the step of inoculating the culture medium recovered in the step b) into the sterilized medium to prepare a new culture medium and re-dispensing the new culture medium into a new tray. The culture solution recovered in the present invention may be directly added to a new tray to produce bacterial cellulose. Alternatively, bacterial cellulose may be produced by inoculating the freshly inoculated medium with the inoculated medium to a new tray.

In the present invention, the culture medium recovered in the step b) is inoculated at 5 to 20% in the sterilized medium. If the amount of the culture solution inoculated into the sterilized medium is small, the amount of the product is reduced accordingly. If the inoculated amount is too large, the cellulosic production may be delayed due to a lack of the medium components.

As described above, the culture solution, which has been terminated after one incubation, is aseptically recovered using the recovering apparatus of the present invention and then cultured in a new tray or inoculated on the sterilized medium, It is possible to produce a larger quantity of bacterial celluloses of better quality and to reduce the waste water treatment cost by collecting the waste culture liquid to be treated for disposal, thereby increasing the production rate of bacterial celluloses.

1 is a sectional view of a bacterial cellulose culture liquid recovery apparatus according to Embodiment 1 of the present invention.
2 is a cross-sectional view of a hydraulic device and a plurality of trays among the bacterial cellulose culture liquid collecting apparatus according to Embodiment 1 of the present invention.
3 is a cross-sectional view of a plurality of trays in a bacterial cellulose culture liquid recovery apparatus according to Embodiment 1 of the present invention.
4 is a cross-sectional view of a liquid recovery line, a valve and a liquid recovery control device in the bacterial cellulose culture liquid recovery device according to the first embodiment of the present invention.
5 is a sectional view of a filter in a bacterial cellulose culture liquid recovery apparatus according to Embodiment 1 of the present invention.
6 is a cross-sectional view of a culture tank and a vacuum pump in a bacterial cellulose culture liquid recovery apparatus according to Embodiment 1 of the present invention.
7 is a view showing a conventional production process of a bacterial cellulose.
8 is a view showing a production process of a bacterial cellulose using the bacterial cellulose culture liquid recovery apparatus of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for illustrating the present invention, and thus the scope of the present invention is not construed as being limited by these embodiments.

Example  One: Bacterial  Cellulose culture liquid recovery device

2 is a cross-sectional view of a hydraulic device and a plurality of trays of a bacterial cellulose culture liquid collecting device according to Embodiment 1 of the present invention, and Fig. 3 is a cross-sectional view of the bacterial cellulose culture liquid collecting device according to Embodiment 1 of the present invention. 1 is a cross-sectional view of a plurality of trays in a bacterial cellulose culture liquid recovery apparatus according to Inventive Example 1;

4 is a cross-sectional view of a liquid recovery line, a valve, and a liquid recovery control device in a bacterial cellulose culture liquid recovery device according to Embodiment 1 of the present invention. FIG. 5 is a cross- FIG. 6 is a cross-sectional view of a culture tank and a vacuum pump in a bacterial cellulose culture liquid recovery apparatus according to Embodiment 1 of the present invention. FIG.

According to the above drawings, the bacterial cellulose culture liquid collecting apparatus 100 according to the present invention includes a plurality of culturing trays 110 for stacking a plurality of layers and culturing microorganisms, a hydraulic device 120 for inclining the culturing trays, , A liquid recovery line (130) connected to each of the culture trays to recover the culture liquid, a filter (140) for removing impurities in the culture liquid in which the liquid recovery lines are integrally connected and storing the culture liquid connected to the filter, A culture tank 150 for culturing, a vacuum pump 160 for moving the culture fluid connected to the culture tank, and a liquid recovery control device 170 for controlling recovery and injection of the culture liquid.

The plurality of culturing trays 110 are mounted on a tray placing table 180 provided with a moving wheel 181 at a lower side corner thereof and a hydraulic device 120 for inclining the culturing tray to a predetermined position, (180). Specifically, the hydraulic apparatus 120 is located on the opposite side of the liquid recovery line 130, and serves to lift a plurality of culture trays 110 by 2 to 3 cm toward the liquid recovery line 130. In addition, the plurality of culture trays 110 are formed with liquid recovery line insertion openings 111 on the side surfaces of each of the plurality of culture trays 110 so that the liquid recovery line 130 for recovering the culture liquid can be inserted. The liquid recovery line 130 inserted in the liquid recovery line insertion port 111 is made of silicon which is resistant to steam treatment for sterilization and the silicone liquid recovery line 130 is bent The culture liquid in the culture tray 110 of the culture dish can be sucked up to the maximum.

A valve 132 for regulating the respective liquid recovery lines is formed in each of the liquid recovery lines 130 connected to the plurality of trays to recover the culture liquid. Specifically, the valve 132 is formed in each of the liquid recovery lines, thereby locking the valve of the liquid recovery line, which is not required to be used, to prevent unnecessary consumption. Further, on the side of the valve, a liquid collection control device 170 for controlling collection and injection of the culture liquid is constituted. Furthermore, each of the liquid recovery lines 130 is integrated into one to be connected to the filter 140.

The filter 140 is formed to be able to remove only impurities that are not cells in the culture liquid. The filter 140 is made of a metal material and has a hole size of about 0.5 to 1.5 mesh. The culture solution having passed through the filter is moved to the culture tank 150 through the integrated solution recovery line 130 again.

A groove 151 is formed in one side of the culture tank 150 for storing and culturing the culture liquid for recovering and injecting the used culture liquid. On the other side of the groove 151, (Not shown).

Comparative Example  1: Using a conventional method without using a recovery device Bacterial  Production of cellulose

As in the production process of FIG. 7, bacterial cellulose was produced using the conventional method without using the recovering apparatus of the present invention, and the period of producing bacteriorcellulose was measured.

As a result, it takes 14 to 15 days to produce bacteriorcellulose once from the seed culture to the end of the stationary culture, and the number of times of producing bacteriorcellulose by about 30 days is about twice.

Example  2: Using the recovering device of the present invention Bacterial  Production of cellulose

The present inventors used the recovering apparatus of the present invention to conduct a stationary culture by increasing the primary secondary culture solution after seed culture as shown in FIG. 8, and after recovering the bacterial celluloses after the completion of the culture, The culture solution collected in one place is collected by using a recovery line and a vacuum pump, the filter is passed through to remove the impurities of the collected fluid, and the culture solution passed through the filter is sterilized And the inoculated medium was divided again to produce bacterial cellulose. The above procedure was repeated twice to produce bacterial celluloses continuously.

As a result, it took 14 to 15 days to produce bacteriorcell cellulose once from the seed culture to the end of the stationary culture, which was the same as that of Comparative Example 1. However, when the bacterial cellulose was continuously produced using the recovering apparatus of the present invention, Since the third culture from culturing to stationary culture can be omitted, the number of times that bacterial cellulose can be produced on the basis of about 30 days was about 6 times that of the comparative example.

100: Bacteriorcellulose culture liquid recovery device
110: Culture tray
111: liquid recovery line inserting port
120: Hydraulic device
130: liquid recovery line
131: Suction port of the liquid recovery line
132: Valve of the liquid recovery line
140: Filter
150: Culture tank
151: grooves for recovering and injecting the culture medium
152: connection groove of the vacuum pump
160: Vacuum pump
170: liquid recovery device
180: Cradle
181: Caster wheels

Claims (14)

A plurality of culture trays (110) stacked in multiple layers to culture microorganisms;
A hydraulic device (120) for tilting the culture tray;
A liquid recovery line (130) connected to each of the culture trays to recover the culture liquid;
A filter (140) for removing impurities from the culture fluid in which the liquid recovery lines are integrally combined and connected;
A culture tank 150 for storing and culturing the culture fluid connected to the filter;
A vacuum pump 160 for moving the culture fluid connected to the culture tank; And
And a liquid recovery device (170) for controlling the recovery and injection of the culture liquid.
The bacterial cellulose culture liquid collecting apparatus (100) according to claim 1, wherein the microorganism produces a polysaccharide secreted out of the cell.
The method according to claim 1, further comprising a cradle (180) provided with a moving wheel (181) at a lower side edge to move the plurality of culturing trays (110), the liquid collection controller (170) (100). ≪ RTI ID = 0.0 > 11. < / RTI >
The bacterial cellulose culture liquid collecting apparatus (100) according to claim 1, wherein the hydraulic device (120) is connected to a lower portion of a tray (180) of a tray.
The apparatus according to claim 1, wherein the tray (110) has a liquid recovery line inlet (111) formed in a side surface of the tray so that a liquid recovery line can be inserted therein.
The bacterial cellulose culture liquid collecting apparatus (100) according to claim 1, wherein the plurality of trays (110) comprise 10 to 60 pieces.
The bacterial cellulose culture liquid collecting apparatus (100) according to claim 1, wherein the liquid recovery line (130) is made of silicon.
The bacterial cellulose culture liquid collecting apparatus (100) according to claim 1, wherein the liquid recovery line (130) is made of a material capable of withstanding steam.
The bacterial cellulose culture liquid collecting apparatus (100) according to claim 1, wherein a valve (132) is formed in each of the liquid recovery lines connected to the plurality of trays.
The bacterial cellulose culture liquid collecting apparatus (100) according to claim 1, wherein the filter (140) is formed so as to remove impurities that are not cells.
The method according to claim 1, wherein the apparatus collects all of the culture liquid from the culture tray, inoculates the culture liquid into the sterilized culture medium, removes the existing plural culture trays, mounts a new plurality of trays, Wherein the culture solution stored in the tank is injected into the cell culture tank.
Method for continuous production of bacterial cellulose using the apparatus of claim 1:
a) preparing a bacterial cellulose using a tray;
b) recovering the cultured liquid from the tray of step a) after the production of the bacterial cellulose by using the culture solution recovery device of claim 1;
c) re-dispensing and culturing the medium of step b) in a new tray; And
d) repeating steps b) and c) three to six times to produce a bacterial cellulose.
[10] The method of claim 10, further comprising the step of inoculating the culture medium recovered in step b) into a sterilized medium to prepare a new culture medium and re-dispensing the new culture into a new tray.
12. The continuous production method of bacterial cellulose according to claim 11, wherein the culture medium recovered in step b) is inoculated to the sterilized medium at 5 to 20%.

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