KR200232168Y1 - the medium animate of air floating balloon - Google Patents

Abstract

The present invention relates to a biocultivator capable of culturing a large number of microorganisms, plant cells.

More specifically, the upper part and the center part have a ball-shaped spherical shape, and the lower part forms a balloon-shaped body formed in a gentle bottleneck shape to maximize the body diameter of the incubator to facilitate the growth of the medium, and A blade is provided at the end of the shaft which is equipped with a lid and penetrates inside the center to prevent bubbles from forming in the process of growing medium and to prevent oxygen from being generated and to prevent generation of harmful substances. It is designed to cut and to supply oxygen at the bottom of the body by installing an air inlet, and the present invention is suitable medium of growth conditions for the bioreactor culture of ginseng, rapeseed, and wild ginseng root muscle regardless of the arable land and season. , Optimal growth conditions such as pH of the medium, sugar concentration, culture temperature, type of growth regulator, etc. In addition, by removing bubbles, harmful substances, and oxygen supply disturbance factors in the growth of adventitious roots, it establishes the optimal requirements for the growth of adventitious roots and industrializes the production of the adventitious roots in the bioreactor of ginseng, which has not been developed until now, without being affected by the climate and environmental conditions. It is to provide incubator to produce value-added ginseng, rapeseed and wild ginseng throughout the year.

Description

The medium animate of air floating balloon

The present invention relates to an incubator capable of cultivating a large amount of microorganisms and plant cells, and more specifically, it is possible to maximize the growth of the root of the root by identifying the factors affecting the growth of the root of the root, such as ginseng, rod, and wild ginseng. The present invention relates to a balloon-type air-floating biocultivator that is capable of multiplying ginseng, rapeseed, and wild ginseng in a large amount regardless of the season.

Ginseng and wild ginseng have been shown to have remarkable efficacy in treating various diseases and promoting the recovery of the disease since ancient times, and extensive research has been conducted to explore the efficacy and pharmacological effects of ginseng. Scientifically proven representative effects are the homeostatic maintenance of body control functions, and based on these actions, anti-fatigue and antistress, antidiabetic, blood pressure control, anticancer, arteriosclerosis and hypertension prevention, brain function Enhancement, gastrointestinal function, immune function enhancement, antiviral action, etc. have been reported.

The main active ingredients of such ginseng are saponins, sapongenins, polyacetylenes, pyrazine derivatives, maltol, and the like.

Natural medicinal ginseng, however, is a non-economic product that is cultivated over a period of four to six years in a well-drained, cool highland and is not only affected by the natural climate, but is also impossible to grow in the same place.

Therefore, plant tissue culture research has been conducted as a method for mass production of expensive medicinal ginseng without disturbing the climate throughout the year, and cultivated root tissues of natural medicinal ginseng to obtain callus, an amorphous cell mass, and various nutritional media and environmental conditions. Research on the production of ginseng tissues containing a large amount of the same active ingredient as natural medicinal ginseng by mass cultivation under the present condition, and at present, the technology for cultivating tissues for various purposes using Korean ginseng, American ginseng, whole ginseng, etc. In particular, many reports have been made in the professional journal about the proliferation from the callus obtained by culturing each tissue of ginseng, organ differentiation, establishment of cell culture from callus, and the like.

However, most of these experiments were mostly related to the analysis of physical and chemical factors on callus and cell proliferation, and there are few experimental results on ginseng root culture of ginseng.

Therefore, the study of intestinal root cultivation is carried out in laboratory incubators used in labs such as culture dishes, and for mass production, it is best to allow the growth of the roots of ginseng, rapeseed, and wild ginseng in large tanks rather than in arable land (land). Ideally, the development of a container that can meet these conditions should be studied together, but it has not been achieved until now, and it is possible to grow by embedding the medium in a large tank. Growing cells secrete persistent growth toxic substances because they do not receive severe drying and nutrient supply, and ultimately, the problem of lowering the vitality of all cultured cells and intertwined roots as they grow, which leads to lower oxygen supply. There was a problem that can not get the root.

The present invention has been devised in view of the above problems, and the upper and center parts are formed into a ball-shaped sphere, and the lower part forms a balloon-shaped body formed in a gentle bottleneck shape, thereby increasing the body diameter of the incubator to maximize the growth of the medium. Smooth and the top of the balloon-shaped body is provided with a blade at the end of the shaft penetrated into the inside of the center to prevent bubbles from forming during the growth of the medium to interfere with the oxygen supply and prevent harmful substances from occurring. In order to supply oxygen during the growth of the medium by installing an air inlet at the bottom of the body, the present invention is suitable for the growth of the bioreactor culture of ginseng, rodworm, and wild ginseng irrespective of the arable land and season. establish optimal growth conditions such as pH, sugar concentration, culture temperature, type of growth regulator, , Removes bubbles, harmful substances and oxygen supply disturbance factors in growing adventitious muscle, establishes optimal requirements for growing adventitious root, and industrializes production of adventitious root in ginseng bioreactor, which has not been developed until now, and is not affected by climatic and environmental conditions. It is to provide an incubator that can produce ginseng root, ginseng root, wild ginseng and other ginseng roots throughout the year.

1 is a block diagram of an incubator according to the present invention.

Figure 2 is a plan view of the incubator according to the present invention.

Figure 3 is a cross-sectional view of the incubator according to the present invention.

Figure 4 is a porous plate installation configuration for a special purpose in the incubator according to the present invention.

<Code Description of Main Parts of Drawing>

1: incubator 2: body

3: lid 4: opening and closing handle

5: motor 6: shaft

7: blade 8: lighting

9: sight window 10: air inlet

11: outlet 12: guide groove

13: Perforated plate 14: Inoculation mouth

15: temperature gauge

Hereinafter, an embodiment of the present invention will be described in detail with reference to the accompanying drawings.

In a culture vessel capable of cultivating a large amount of plant cells.

A balloon-shaped body (2) formed in a ball-shaped spherical shape at the upper and center portions and a lower bottleneck at a lower portion thereof;

A shaft (6) having a lid (3) having an opening / closing handle (4) at an upper end of the balloon-shaped body (2) and penetrating therein;

A blade (7) installed at the upper end of the shaft (6) and rotating at the lower end by driving the motor (5);

A plurality of lighting windows 8 formed on the upper surface of the body 3 around the lid 3 to allow light to pass therethrough;

One side of the balloon-shaped body (2) and a plurality of viewing windows (9) formed to observe the growth process of the root muscle;

A temperature gauge 15 installed to check the temperature of the inoculation port 13 and the temperature to allow the medium to be injected into one side of the bottom of the wind-type body 2;

The bottom of the body (2) is a configuration of the intestinal root incubator (1) provided with an air inlet (10) and outlet (11).

In addition, as another embodiment of the present invention by forming a guide groove 12 on the inner side of both sides of the lower 1/4 point inside the balloon-shaped body (2) to install a porous plate 13 to grow other media other than ginseng para muscles Configuration. Reference numeral 4 'is a fixture supporting the lid opening and closing handle shaft, 16 is the bioculture vessel support leg, 17 is the pressure gauge.

In order to achieve the object of the present invention as described above, a description will be given of a manufacturing process for growing a medium (culture medium) into a root muscle and mass production prior to describing the air incubation bioculture device.

Mass growth method of ginseng, chungsam ginseng and wild ginseng root of ginseng by tissue culture according to the present invention is to induce the callus by culturing the tissues such as roots, stems, leaves, etc. Generating and growing the negative root from the callus, and culturing the grown root in a bioreactor to grow in the method of mass propagation of ginseng, tortoise and wild ginseng.

Soy root growth is carried out in this way is to enable the production of pollution-free ginseng irrelevant to the environment, soil and pesticide contamination throughout the year in the laboratory or factory using the air-cultured bioculture device of the present invention, the embodiment will be described below. do.

First, about 80% is added to the volume through the inoculation port 13 provided at one side of the lower center of the incubator 1. For example, in the case of a tank having a volume of 500 cc, the medium (culture medium) is added at a ratio of about 400 cc (medium loading: container 100: medium 80 ratio). At this time, the capacity of the incubator 1 may be optionally provided as 1 ton, 2 ton, 3 ton ‥‥‥ 10 tons, 20 tons 30 tons, etc., the medium is sterilized in a steam container at 121 ℃ for about 40 minutes Through the inoculation port 13 of the growth incubator (1) through.

When the medium (culture medium) is introduced into the incubator as described above, the air is continuously supplied by continuously injecting oxygen through the air inlet 10 formed at one side of the bottom surface of the body 2.

The inside of the incubator (1) maintains the optimum temperature 23 ~ 25 ℃ the medium can be grown as a root muscle and the temperature inside can be confirmed by the temperature gauge installed on one side to maintain the temperature inside.

When the medium (culture solution) is added and temperature and oxygen are adjusted as described above, bubbles are generated as the medium in the biocultivator grows, bubbles rise and rise above the water surface in the incubator, hit the inner wall, and the bubble traces thereof are dried. It may be changed to harmful substances that hinder the growth, but at this time, the shaft 6 penetrating the central portion of the lid 3 of the growth incubator 1 is rotated every predetermined time by the driving of the motor 5. The lower end of the shaft 6 has a blade (7) attached to the blade (7) is rotated while stirring the medium to ensure that the oxygen supply is made smoothly as a whole and bubbles are generated in advance to prevent the formation of foam bubbles It prevents it from drying by hitting the inner wall and transfers the root muscle to the secondary incubator to further grow and cut into 2 ~ 3㎝ length.

Under the above conditions, the medium is grown into the root muscle during the culture period of about 40 days.

In addition, the growth roots in the growth culture can be observed through the duct through the viewing window (9) formed on one side of the body (1), the oxygen supply, temperature can be adjusted according to whether the observation, the blade ( 7) Rotate to evenly change the location of the multiplying the roots of the proliferation and at the same time prevent the intertwined roots intertwine to prevent growth.

The outer periphery of the upper lid (3) of the present invention is provided with a plurality of illumination windows (8) to transmit the light necessary for the growth of the medium.

As described above, if the culture medium of the incubator is grown to the intestinal muscle after about 40 days and completely grows, it produces the root muscle. Before discharging the root muscle, the outlet 11 installed in the center of the bottom of the body is opened to completely discharge all the liquid in the incubator. After discharging, the upper opening handle 4 is rotated so that the shaft of the opening handle 4 is lifted by the fixture so that the lid 3 can be opened, and the raised lid is pulled to one side to completely open the growth incubator. At the same time, by opening the lower end of the body (2) it is taken out of the growing root muscles.

Looking at the experimental results studied to propagate the roots produced as described above are as follows.

In this experiment, 6-year-old ginseng, 15-year-old Janggesam, and 100-year-old wild ginseng were used in the following experiment.

Example 1

Experiment for selecting optimal condition for paraparesis

Forming the root muscle from callus and then maximizing the growth of these tissues within the shortest period of time was the most efficient way to reduce production costs. Therefore, experiments were conducted to identify the optimal conditions as follows.

30 ml of MS medium to which IBA 2.0 mg / L was added was dispensed into a disposable petri dish, and 30 g of intestinal roots derived from callus of wild ginseng were cut to an average length of 10 mm.

Experiments were carried out at 1, 1/2, and 3/4 times the concentration of minerals in the medium. The pH of the medium was divided into 4.0, 5.0, 5.5, 5.7, 6.0, 6.5, and the sugar concentration in the medium was 0%, Divided into 1%, 2%, 3%, 4%, 5%, 6%, and 7%. The culture temperature was divided into 15 ℃, 18 ℃, 20 ℃, 22 ℃, 24 ℃, and 26 ℃ to observe the growth. It was.

As a result of the experiment, it was found that the range of inorganic concentration 1/2 to 3/4, medium pH 5.7 to 6.0, medium sugar concentration 3 to 5%, and culture temperature 18 to 24 ° C. were the optimum conditions for the growth of the root muscle.

Example 2

Effect of Growth Regulators on the Growth of Abdominal Muscle

After adding 3% of sugar to MS medium and fixing pH to 6.0, IBA, NAA, and BSAA were each added 1 mg / L, 2 mg / L, 3 mg / L, and 5 mg / L, respectively. The base medium was dispensed into 1 L Erlenmeyer flask and 5 L bioreactor, respectively, and then inoculated with the roots of wild ginseng and cultivated ginseng (average 1.0 cm in length), and the growth was examined after 4 weeks, and the results are shown in Table 1.

As can be seen from Table 1, the growth without the growth regulator showed less than two-fold increase, whereas the medium added with IBA and NAA showed a five-fold increase in biomass, showing the most effective results at 2-3 mg / L. It was found that the concentration is appropriate. On the other hand, the addition of BSAA, a newly synthesized oxine, significantly increased the body weight and the titer was also found to be 2-3 mg / L.

Comparing the live weight of cultivated ginseng and wild ginseng, there was no difference in growth rate between cultivated ginseng and wild ginseng in the bioreactor.

Therefore, it was judged that addition of growth regulators such as NAA, IBA, and BSAA was recommended for mass growth of ginseng root muscle in the bioreactor.

Example 3

<Influence of Inoculation Method on Growth of Abdominal Muscle>

In order to examine the effect of the preparation method of root muscle on root proliferation when culturing wild roots of wild ginseng and cultivated ginseng in a bioreactor, uncensored culturing was performed on MS medium supplemented with BSAA 2mg / L, except for fragments. After culturing for 4 weeks by dividing the length of the adductor in 1 ~ 2 ㎝ without culturing the method, section and aide root, the results are shown in Table 2.

As shown in the results of Table 2, when the aides formed in the explant culture were transplanted into the fresh medium together with the explants, the entrainment did not occur, so that the weight gain did not occur and the browning phenomenon occurred. On the other hand, when the original fragments were removed and only the formed apex was cut and cultured, the abundance incidence and live weight increased, but the inoculation took a long time and was not an effective method.

However, the newly formed aides, including culture fragments, were randomly cut by 1 to 2 cm, and the growth rate was 15 times higher when inoculated into fresh medium, which was most effective for the increase of the live weight. Therefore, it is preferable to inoculate by cutting the length of the adductor 1 to 2 cm without distinguishing between sections and aides.

Example 4

Effect of Bioreactor Morphology on Abdominal Muscle Growth

When cultured using a bioreactor for the purpose of mass propagation, 40 days after incubation in a balloon-type air-lift bioreactor (1) of 500 L and 1000 L, which is a pilot scale, to observe the results of the growth of the root muscles according to the shape of the bioreactor The effect on the increase in live weight was examined.

At this time, the temperature of the culture chamber was fixed at 22 ° C., and the air injection amount in the bioreactor (1) was 0.05˜0.3 vvm. The medium used MS medium added with 3% sugar and BSAA 2.0 mg / L was added as a growth regulator. And pH was adjusted to 6.0.

Effect of Bioreactor Type on Mountain Gastrocnemius Muscle (40-day Culture) Type of incubator Scale (ℓ) Initial inoculation amount (kg) Live weight (kg) Under building (㎏) Air Floating Balloon Incubator 500 2 88 6.5 1000 3 185 12.4

As can be seen from the results of Table 3, the air flotation balloon type was effective in increasing the weight and dry weight. However, as the culture seal elapsed, entanglement of the root muscles occurred, thus increasing the air injection amount every two weeks to prevent root entanglement. In order to prevent a decrease in the total production due to the floating phenomenon of the root muscles occurring after 4 weeks of culture, it is preferable to reinoculate (about 1 kg) after 2 weeks of culture.

Therefore, in the 1 ~ 3ton scale-up incubator, which is the preliminary growth incubator for operating 50L, 100L seed and incubator of 20 ~ 50ton, the blade (7) using the motor (5) is attached and transferred to a larger incubator. It is preferable that the previously formed adductor roots are cut to a length of 2 to 3 cm by the method of Example 3 and then transferred.

As seen above, the mass growth method of ginseng, chungsam ginseng and wild ginseng root of the present invention was confirmed that it is a breakthrough mass production method with high biomass production capacity, easy handling, and increased building weight.

The internal structure of the growth incubator of the present invention should be formed in a structure that can prevent contamination, and for harvesting and internal cleaning after cultivation, one side plate of the incubator has a case structure and is configured to be operated by mechanical and automatic means.

In addition, oxygen supplied through the air inlet 10 of the bottom of the incubator 1 body 2 should pass through a 0.2 μm filter, and a sparging metal filter (0.2 to 5 μm) mounted on the lower part may provide sufficient air for culture. It must have an area that can pressurize, and the thinner the particles of air, the better it is to grow.

The structure of the air supply device should be suitable for plant cell culture, and the metal filter should be a structure that is easily disassembled and assembled and is not leaked.

A bed plate for bronchial support is installed inside the incubator, which must be opened and opened for disassembly, control of the internal pressure of the incubator during cultivation, and in the vent to prevent contamination during unavoidable blocking of compressed air. A filter must be attached and an air storage tank is required for this purpose.

When sterilizing steam inside the incubator, the internal pressure must be maintained manually. The inside of the incubator can also sterilize the connecting pipes and accessories. Steam sterilization is carried out at 121 ℃. The incubator is equipped with temperature, pressure gauge, pH sensor and DO sensor.

Growth incubator of the present invention made as described above is not necessarily used only for the growth of ginseng, wild ginseng, thoracic ulcerative root muscle, it is possible to apply to other plant cell culture as needed.

Regardless of the location and season of the present invention, it is possible to mass-produce ginseng roots, wild ginseng, and inferior root muscles, which are superior to the effects of ginseng, wild ginseng, and rapeseed in a short time, and to mass produce wild ginseng, which is highly recognized by consumers. It is designed to meet the consumer's consumption needs.

In this way, the growth culture device of the present invention enables mass production of gross roots of ginseng, wild ginseng, elders, etc. in a short period of time, thereby providing high value-added gross roots. It is possible to suppress the formation of harmful substances, and to cut off the tangled muscles with the growth of the roots of the roots with a knife to cut smoothly the supply of oxygen and transfer the formed roots of 2 ~ 3㎝ length before transferring to the larger incubator. In particular, it was possible to provide conditions, and in particular, it was possible to establish the commercial root production of ginseng in the incubator, which has not been developed until now, and by cultivating wild ginseng, it can be supplied at low prices to various demand groups.

Claims (3)

In a culture vessel capable of cultivating a large amount of plant cells. A balloon-shaped body (2) formed in a ball-shaped spherical shape at the upper and center portions and a lower bottleneck at a lower portion thereof; A shaft (6) having a lid (3) having an opening / closing handle (4) at an upper end of the balloon-shaped body (2) and penetrating therein; A blade (7) installed at the upper end of the shaft (6) and rotating at the lower end by driving the motor (5); A plurality of lighting windows 8 formed on the upper surface of the body 3 around the lid 3 to allow light to pass therethrough; One side of the balloon-shaped body (2) and a plurality of viewing windows (9) formed to observe the growth process of the root muscle; A temperature gauge 15 installed to check the temperature of the inoculation port 13 and the temperature to allow the medium to be injected into one side of the bottom of the wind-type body 2; Balloon type airborne bioreactor, characterized in that the bottom of the body (2), the air inlet 10 and the outlet 11 is provided with a root muscle incubator (1) is installed. The method of claim 1, Balloon type airborne organisms, characterized in that the guide grooves 12 are formed on both inner side walls of the lower part of the balloon-shaped body 2 to install a porous plate 13 for plant cell culture. Incubator. The method of claim 1, Oxygen supplied through the air inlet 10 at the bottom of the bio-incubator 1 body 2 passes through a 0.2 μm filter, and a sparging metal filter (0.2 to 5 μm) mounted at the bottom is sufficient for culture. Balloon type air culture bio-cultivator, characterized in that to be able to exert air pressure.