WO2007045125A1

WO2007045125A1 - Method and system of storage-type tissure culture or microbe fermentation

Info

Publication number: WO2007045125A1
Application number: PCT/CN2005/001733
Authority: WO
Inventors: Pai-Cheng Chen
Original assignee: Pai-Cheng Chen
Priority date: 2005-10-21
Filing date: 2005-10-21
Publication date: 2007-04-26
Also published as: CN101292038A

Abstract

Method and system of storage-type tissure culture or microbe fermentation which was the design that combined the food manufacture automation technique with the sterile room technique, the automated storage technique and the microbe fermentation technique. To match the design of the sterile room space, after the media were putted into the fermentation vessels with the automated mechanism and the sterilization procedure was accomplished, the strains were inoculated, the fermentation vessels were placed onto the shelf of the storage area to storage with the automated access operation, there were growing monitor devices, stirring devices, shaker devices, separate air or oxygen supplying devices on the each shelf to provide all the required for the strains or cells, other microbes during the fermentation.

Description

Warehousing tissue culture or microbial fermentation method and system

The invention relates to a storage tissue culture or microbial fermentation method and system, in particular to a design combining food manufacturing automation technology, sterile room technology, automatic storage technology and microbial fermentation technology, and the integrated technology is used in tissue culture, The liquid, semi-solid, solid-state fermentation process of microorganisms, using the storage principle, to advance the fermentation industry and tissue culture to a technological innovation of mass production.

Background technique

Since ancient times, humans have learned to use warehouses to store items, set up shelves in warehouses to make more efficient use of space, and cooperate with other protective measures to prevent stored goods from reaching rodents, insects, ants, moisture, and sputum to increase storage time. Moreover, with the advancement of technology, mechanical automatic control technology is becoming more and more mature, and automated warehousing has emerged. This not only makes space more efficient, but also reduces personnel utilization and increases work efficiency due to automatic access to equipment. However, the current automated warehousing system is only used to store goods, but the management work does not stop for a moment. These costs will increase the cost of the goods and will be passed on to the consumers. If the logistics is delayed, the loss will be more difficult. estimate. For this reason, if the convenience provided by automated warehousing can be effectively utilized and the storage time is effectively applied, the waste of space costs can be reduced, and the utilization of automated warehousing can be further improved. Among them, tissue culture and microbial fermentation engineering are the areas of work that make full use of automated warehousing. Because these tasks require time, space, and proper environmental control, if they can effectively use automated warehousing for management, they will be able to maintain quality and reduce Product cost, available to those who need it at a low price.

Humans have long known to use fermentation to make the food or alcohol they need, such as whole grains or fruit making, taro, bread fermentation, tea fermentation (black tea, Pu'er tea), soy sauce made with soybean meal, red yeast, etc. These are all based on solid-state fermentation or semi-solid fermentation, so humans have long known how to use solid-state fermentation methods to make the food they need.

However, the above traditional fermentation methods have several common points:

1. The fermentation fungi are mostly yeasts or molds;

2. Fermentation at normal room temperature and normal conditions;

3. Mostly, the medium (fermented product) is filled in a disc vessel or a shallow dish container;

4. The fermentation space does not need to be specially sterilized;

5. Let the bacteria be ventilated and heat-dissipated when the amount of the medium is large or thick. Normal fermentation grows.

Traditional solid-state fermentation methods are difficult to control in terms of quality control. The quality, heat and gas transport in fermentation cannot meet the required standards. For example, the production of koji is not only the production of yeast and aspergillus, but also some acid and odor. Sexual bacteria, such as esterified bacteria, lactic acid bacteria... The production of koji is the use of conditional control to screen high-quality yeast and aspergillus in the natural environment, but it uses almost unsterilized raw materials. The growth is also in the curved room. The air is circulated with the outside world, so in the production of koji. In the process, not only the production of yeast and Aspergillus, but also some acid-tolerant and anaerobic bacteria, there is room for improvement in the traditional solid-state fermentation method.

In modern times, many fungal products are fermented by mycelium. In the fermentation process of mycelium, it is generally divided into liquid fermentation and solid fermentation. In the liquid fermentation method, when the fermentation amount is small, the container is mostly a triangular flask. Large-scale fermentation uses a large fermentation tank; in the solid-state fermentation process, the container used is a culture flask or a drum stirred fermentation tank. The culture bottle method requires more disinfection procedures, consumes too much labor, and has less fermentation. The stirred fermentation tank is limited by the heat dissipation and ventilation problems of the solid medium, and the yield of single fermentation is not large; and for the continuous heat dissipation of the medium, the rolling must be continuously stirred, the hyphae are continuously split, and the strain is quiet because it cannot be allowed to stand. The growth, so the strain is less mature, so there is a problem of less production of secondary metabolites.

In recent years, under the efforts of industry and academia, a variety of rotary agitated solid-state fermentation tanks have been developed, which can effectively control the growth requirements of temperature, humidity, and aeration during the growth process, but the production volume has not been able to increase due to heat dissipation. Can not effectively overcome, if the amount of the medium is too much, the internal temperature rises, causing the strain to die and the fermentation to fail. Such a fermentation tank rotates and stirs the medium in the tank to achieve the purpose of aeration, heat dissipation and mass transfer. However, as the bacteria grows well, as long as the aeration is good and the heat dissipation is good, excessive agitation may affect the growth of the bacteria. Another disadvantage of the type of fermentation tank is that if the rotation and stirring are stopped, although the bacteria can be quietly grown, heat dissipation and ventilation problems occur, so that the fermentation product cannot be mass-produced.

At present, tissue culture and microbial liquid fermentation are large-scale production. The medium is in the fermentation tank for the purpose of aeration and mass transfer by means of rotary stirring or aeration mixing. The advantage is that the fermentation space can be designed and produced almost infinitely. The disadvantage is that during the fermentation process. If there is intrusion of bacteria to cause infection, it is difficult to detect in the process. Although it is possible to judge whether the bacteria infection is caused by observing the change of pH value, some bacteria infection will not change the pH value, and it is necessary to wait for the output to be tested. Analysis; For some fungi and some special output requirements for special metabolites, it is not suitable for liquid fermentation. Because liquid fermentation has a short growth period, many secondary metabolites of fungi cannot be produced because the strain is not mature enough. . And after drying, there is not much solid product.

Solid-state fermentation is currently available in both manual and semi-automatic ways. Manually, the medium is placed in a bottle container. The capacity is about 200~400 kg. If the medium is too large, it will cause serious heat dissipation and ventilation problems. The culture bottle has no stirring and unnatural air intake device. After the medium is cooked and cooled, it is loaded. Into the culture bottle, then inoculate after high temperature sterilization, cover the cap with the vent filter hole, and then put a certain number of culture bottles in a temperature-controlled space, let the bacteria grow in the bottle, but because there is no stirring Mandatory ventilation equipment, and the inoculum (or granules) are only distributed on the surface above the medium, so the quality of the medium must be placed on the oscillating machine at a specific time to spread the strain between the culture medium in a turbulent manner. , to achieve the purpose of long-term bacteria in each medium. The ventilation method is to make the air in the bottle and the outside of the bottle convect naturally by a simple filter cloth or a filter screen at the mouth of the bottle cap, the ventilation effect is poor, and the infection is easily caused by the entry of the bacteria, thereby reducing the excellent rate of the product.

In addition, in the existing mushroom mushroom cultivation process, some mushrooms are currently cultivated in a shelf manner, and the space capsule (solid matter culture package) is generally used to cultivate the fruiting bodies of mushrooms and ganoderma lucidum, and to cultivate a space environment. The management only has conditions such as temperature, humidity, light, etc., and the aseptic management of the cultivation environment is still not available, and the automated storage facility has not yet been applied to the fruiting body cultivation. The mycelium culture remains in the liquid fermentation tank, the culture flask and the small (500 liters or less) solid state fermentation tank stage. Therefore, the prior art is difficult to maintain the quality and output of the product, so that the price of the functional food remains high.

In the current fermentation industry, the cultivation of mycelium has two common methods of liquid fermentation and solid state fermentation. In the industrial production of microorganisms, large-scale fermentation tanks are used for liquid fermentation, while solid-state fermentation uses culture flasks or small solid-state fermentation tanks. Although the culture bottle can also be stored in a large amount by the shelf method, the purpose is to increase the storage capacity of the culture bottle in a certain space, but the shelf itself has no equipment such as stirring, vibration, ventilation and growth monitoring, and the conventional storage space lacks the aseptic space management. The equipment is set up, so the entire operation process takes more time and labor, and the infection rate of the bacteria is high, making it difficult to perform high-quality, high-efficiency mass fermentation applications.

The invention is to apply the technology of automatic storage in tissue culture and microbial fermentation engineering. The previous storage system is only used for the storage of raw materials, semi-finished products or finished products, and does not have the function of manufacturing. The invention adopts the management technology of the aseptic space, plus the shelf and storage space with the functions of stirring, vibration, ventilation, temperature, humidity, pH control and growth monitoring, and the management capability of the fermentation condition plus the design of the shelf space. A variety of fermentation tanks, as well as automated storage equipment, constitute a new automatic storage tissue culture (tis sue cul ture) and microbial fermentation system. Therefore, the biggest difference from the previous storage equipment is that the previous storage is only a place to store goods. The present invention is to make the storage system no longer a warehouse, but a manufacturing plant, a high-capacity production equipment for tissue culture and fungus fermentation.

Summary of the invention The object of the present invention is to provide a storage tissue culture or microbial fermentation method and system, which is a combination of food manufacturing automation technology, clean room technology, automated storage technology and microbial fermentation technology design, and the integrated technology is used in Tissue culture, microbial liquid, semi-solid, solid-state fermentation process, using the storage principle, the fermentation industry and tissue culture to a large-scale production and manufacturing of a technological innovation.

In the past bacteria process, mold and yeast are strong bacteria in the environment and are common in the natural environment. Therefore, the past fungus products such as koji, cheese, soybean meal, black tea, Pu'er tea, red yeast rice, etc. Etc., using environmental control to screen and cultivate high-quality mold or yeast, the air in the fermentation space is circulated outside, so there will be a little microbial colony on the medium, but the temperature and humidity at that time are for the bacteria. It is set, so the bacteria is a strong strain, and it will grow vigorously. Other bacteria will become weak or few colonies in this environment, but it will still affect the quality of the fermentation process.

When the medium is sterilized in a solid medium, if the internal ventilation and heat dissipation conditions of the medium are good, no need to stir or flip, so that the hyphae naturally grows, and after becoming a mature strain, the secondary metabolites should be produced. In the process, if you need to stir and turn, it is mainly to let the strain be fully distributed in the medium, and the purpose of ventilation (no need for anaerobic bacteria) and heat dissipation. If the three elements are all available, the medium can be allowed to stand still. The bacteria grow normally.

With the improvement of modern sterilization equipment and the maturity of automation equipment, the process of natural ventilation in the past can be effectively improved, the fermentation quality is more stable, the output is larger, and the production scale is unprecedented in the solid state process.

Solid-state fermentation is the most suitable method for artificial cultivation of fungi. It has been widely used in the cultivation of fruiting bodies, such as the cultivation of mushrooms, Ganoderma lucidum, Brazilian mushrooms, Pleurotus eryngii, etc. in space bags, but in the cultivation of mycelium, solid state Fermentation faces many bottlenecks, it is impossible to mass produce and lacks effective sterile space management, and it is easy to infect bacteria, resulting in low production rate.

In the air, there are most common bacteria, molds and yeasts. In the natural environment, they are strong bacteria, and edible fungi that are artificially cultivated, such as Cordyceps sinensis, Antrodia camphorata, mulberry yellow and other edible fungi of high economic value, their native habitats. The growth environment is different from the general environment of the flat land. If the cultivation environment is not properly managed, the bacteria to be cultivated are easily infected by the invasion of other bacteria, and the fermentation cultivation fails. This situation is an urgent problem to be overcome in the industry.

The invention utilizes a modern delta dynamic storage technology, cooperates with a sterile room space design, and automatically implants the culture medium into the fermentation container and completes the sterilization process, and then implants the bacteria, by an automatic access machine (or conveyor belt, Other handling machinery) Place the fermentation container on the shelf in the storage area, and each shelf has Growth monitoring devices, agitation devices, vibrating devices, separate air or oxygen supply devices provide all the bacteria needed during the fermentation process.

If such a fermentation vessel is applied to a solid medium, the best design is a flat disk fermenter, the purpose of which is to increase the volume of the medium in the fermentation vessel without affecting the ventilation and heat dissipation of the medium, and to use a stirrer. Or by placing a ventilating heat-dissipating partition in the tank, dividing the medium in the tank into several zones, increasing the effect of ventilation and heat dissipation, and the shape may be circular, square or other specially designed shapes; if applied in liquid and semi-solid In the process, the fermenter generally has no shape limitation, but it is necessary to install a temperature control tube in the fermenter to cool or increase the temperature (if the fermenter is made of metal, the heat transfer is high, and the temperature control of the fermentation chamber can also be used to indirectly Temperature control of the fermenter). The fermentation vessel is internally provided with a stirring device, or the ventilation medium is used to partition the medium into a number of culture zones, and the separator must have both heat dissipation and ventilation functions; visible growth characteristics of the bacteria can be used for natural air in ventilation. Convection or direct introduction of air or oxygen. Of course, a certain number of culture bottles can also be placed on the container and sent to the shelf of the storage area by the automatic access machine, but this method is more labor-intensive, and the infection rate of the bacteria is high, and the production efficiency is poor. DRAWINGS

The following detailed description of a preferred embodiment of the present invention and the accompanying drawings will further illustrate the technical contents of the present invention and the purpose of the present invention;

Figure 1 is a structural diagram of the system of the present invention;

1A is a diagram showing a variation of a shelf storage cell of the present invention;

2A is an enlarged view of a structure related to a fermentation tank of the present invention;

2B is a structural view of another fermentation tank of the present invention;

Figure 3 is a general flow chart of the present invention;

Figure 4 is a flow chart of direct fermentation of a high pressure culture dish according to the present invention.

In the figure, the main component symbol description - 1 a storage space, 2 - automated storage system, 3 - shelf (warehouse), 31 - storage,

32—bearing platform, 33—monitoring device, 4-sterile air system, 5—fermentation tank (bioreactor), 51 a—separator, 51b—separator, 52—stirring blade, 6—motor, 61—strength magnet. Detailed ways

Please refer to FIG. 1 and FIG. 2, and the storage type tissue culture or microbial fermentation system referred to in the present invention mainly includes - A storage space 1, designed and built according to the specifications of the clean room, with various sterilization equipment, such as high temperature and high pressure sterilization, plasma sterilization, ultraviolet sterilization, etc., so that the space can be maintained before fermentation and during fermentation. Aseptic state

An automated storage system 2 having a storage structure, a storage/removal machine, a removal and storage workstation, a material handling and conveying device, a medium conditioning, an inoculation, a filling automation device, all of which are integrated with tissue culture and microbial fermentation, Manufacture

A shelf 3 (also known as a storage rack) is made of high-capacity, oxidation-resistant, high-temperature resistant, and corrosion-resistant materials, and provides a storage compartment 31 for storing the fermentation tank 5 described later. The storage compartment 31 is designed according to the storage space 1 plan. In order to achieve maximum space utilization and convenient control, a loading platform 32 is disposed at the bottom of each storage compartment 31, and the loading platform 32 is used to provide a housing space for other functional devices. If the static storage is used, the storage compartment 31 can carry the platform. 32 or other carrying means to provide the required space for the fermenter;

The load platform 32 of each shelf 3 of the shelf 3 is provided with an intake pipe and a joint, and the fermentation tank 5 placed in each of the storage cells 31 is connected, and the sterile air is transported into the fermentation tank 5 for use by the strain; One embodiment is that the air in the fermentation tank and the air in the clean room are naturally convected without passing through the intake line;

The carrying platform 32 of each shelf 3 of the storage compartment 31 is provided with a vibration device or a rotating stirring device, so that the medium in the tank can produce a sufficient quality transmission effect, or a rotary stirring device can be arranged on the fermentation tank 5, or a gas lifting device can be provided. The method achieves the quality transmission and ventilation effect; the other embodiment provides that the storage compartment only provides the fermentation tank placement space, and the static placement method allows the microorganisms to grow naturally;

The shelf 3 is equipped with a monitoring device 33 for temperature, humidity, pH, oxygen content, etc., to maintain the optimal growth environment of the bacteria in the fermentation tank 5; temperature, humidity, pH value and The relevant monitoring device can also be on the shelf or in each fermentation storage area;

Referring to Fig. 1A, the foregoing shelf 3 may be provided with a hanging platform 32, but is provided with a hanging plate 32a, and is suspended on the hanging plate 32a by a movable storage compartment 31a, so as to be conveniently placed into the shelf 3 by being exchanged.

a sterile air system 4 that maintains the air entering the storage space 1 and the fermentation tank 5 described later in a sterile state;

A fermentation tank (not shown) is placed in a breeding area, mainly in a liquid fermentation process (or solid state), and the inoculum (or inoculum) with the number of colonies reaching the set standard is implanted. In the culture medium of the fermentation tank in the storage tank described later, the seed solution is inoculated into a larger amount of liquid or solid medium through the inoculating hole of the g seed or the fermentation tank;

a fermenter 5 (also known as a bioreactor), the universal type is designed in a flat shape, and the shape is circular and positive. Square and rectangular; the medium in the round fermentation tank 5 is mainly composed of rotary stirring, and the square and rectangular shapes are mainly vibration. The ventilation heat-dissipating partitions 51a and 51b are arranged in the tank, and are divided into general-type grids 51a. And the hollow type grid 51b, the solid medium is divided into several zones, the function of which is to maintain the medium for good heat dissipation and ventilation, and the tunnel type can be rotated or vibrated.

The foregoing system is in a state where the medium is in a solid state, and if the medium is liquid or semi-solid, rotary agitation is the best quality transfer method. Although aeration agitation is also a useful method, if the cultured microorganism is anaerobic or does not require a large amount of air, the aeration agitation method is not suitable. The ideal design of the fermentation tank 5 mixing function, if the power is transmitted from the bottom, and the motor 6 and the bearing drive the stirring blade, the contamination of the oil seal gap is very likely to contaminate the fermented material, and if there is leakage, it will contaminate the fermented material and the fermentation tank 5 . Therefore, if the power is transmitted from the bottom of the fermentation tank 5, the bearingless design should be used, and the rotor driven by the motor 6 is installed in the carrying platform 32 of the shelf storage compartment 31 by the principle of magnetic "same-repeat and opposite-sex attraction". The gear, the runner or the gear unit has a number of powerful magnets 61, and the agitating blades 52 in the fermentation tank 5 also install a number of powerful magnets. When the lower motor 6 rotates, the stirring blades 52 in the fermentation tank 5 are driven to follow. Turn and stir the medium in the tank.

The power source can be driven by the motor 6 or by a separate gas circuit, using a pneumatic machine to drive the wheel, and then the wheel or gear drives the rotation of the agitating blade 52 in the fermentation tank 5.

The power can also be transmitted from above. Since there are vents, inoculation holes, sampling holes, observation holes and related facilities (not shown), in order to increase the proper distribution of the space above, the power source should not be designed from above. In order to increase the space efficiency and reduce the equipment cost, and avoid the configuration of one motor 6 per fermentation tank 5, several runners or gears should be driven by the motor/belt to save space in the storage compartment 31.

The aforementioned fermenter 5 differs according to its function:

a, general fermentation tank

After the medium is inoculated, it is placed in a fermentation tank for fermentation, or the medium is placed in the fermentation tank and then inoculated. The tank has a stirring blade or a ventilating heat-dissipating partition to allow cells or microorganisms to be in the tank under suitable cultivation space and environmental conditions. Internal growth

b, compound fermentation tank

The structure is similar to that of a pressure cooker (fast pot). The medium is sequentially sterilized, steamed, cooled, inoculated, and fermented in a pot. The tank cover has a pressure relief hole and a safety valve in addition to high temperature cooking, and more inoculation holes, Sampling hole and monitoring device: Install the stomata hole in the tank cover or the waste to provide cells or microbial growth. If it is an anaerobic microorganism, the entire tank can be sealed.

If a solid medium process is used, the fermentation tank should be designed in an internal and external tank, and the medium should be placed in the inner tank. The outer tank is filled with water or other liquid, the bottom of the tank is heated by a heat source, the medium is treated by sterilization, and sterilized, and then inoculated after cooling. The groove cover can also be designed with inner and outer covers. The inner cover is made of air filtering effect and ventilation heat dissipation effect. When the high temperature and high pressure cooking is completed, the outer cover can be removed, and the inner cover is provided with a large ventilation heat dissipation area. The natural ventilation mode provides a better growing environment for cells or microorganisms. The same effect can be achieved if the boiler steam is introduced into the fermentation tank for cooking and sterilization.

If the air is introduced into the tank by an unnatural external force, it can be designed with a single cover. The air inlet or the vapor pressure relief hole of the upper cover becomes the air outlet.

In the above solid medium or liquid medium, the solid medium mainly provides the carbohydrate (carbon source) required for the growth of microorganisms, and the selection thereof is rich in carbohydrates and related nutrients, and has good moisturizing. It is also important to consider the cost, the water resistance of the impregnated water, the non-sticking permeability between each medium, and the resistance to agitation.

Monosaccharides and polysaccharides as well as starch can provide the carbon source required by the present invention, and the starch is mainly composed of whole grains, such as coix seed, sorghum, millet, barley, buckwheat, beans, potato (sweet potato, potato). , can be a solid medium material; because starch must be saccharified to become a microbial nutrient, therefore, in the pre-treatment of solid medium, the appropriate amount of sugar must be given to provide the initial carbon source of microbial growth need.

Moreover, according to most studies, the composite carbon source (complex composition and rich in cellulose, hemicellulose) is conducive to the growth of mycelium and the secretion of bioactive substances, the reasons may be as follows, the composite carbon source contains other Nutrients, such as various vitamins and trace elements, of which cellulose is a precursor of the synthesis of mycelial extracellular polysaccharides, which in turn promotes the synthesis of extracellular polysaccharides. Animal experiments have found that coix seed has the effect of lowering blood fat, also contains quite rich water-soluble fiber, and very rich B-type vitamins (such as vitamins Bl, B2 and B3), so it is a very good solid medium, which helps bacteria Deep fermentation of filaments and formation of biologically active substances.

The nitrogen source is an indispensable raw material for the synthesis of proteins, nucleic acids and metabolites by microbial cells. Bacterial cells cannot use inorganic nitrogen to synthesize certain essential amino acids. Therefore, organic nitrogen is the most suitable nitrogen source for microbial growth. The fermentation effect of the composite organic nitrogen source is better than that of a single organic nitrogen source, such as yeast powder, yeast extract, protein, bean cake powder, peanut cake powder, rice bran, wheat bran, etc., and the fermented material has high bacterial content. Fermentation from a single nitrogen source.

Vitamin B, especially vitamin B1 (thiamine), is an essential vitamin in the growth process of the body cells. Adding vitamin B1 to the medium helps the growth of the bacteria. If vitamin B1 is lacking, the bacteria will grow slowly or stop growing. Vitamin B (L-Carni t ine ) can promote the growth of bacteria and increase the resistance to impurities. Bacteria and the environment are resistant to stress.

Supplemented with magnesium (Mg) and potassium (K) and phosphorus (P) compounds, which are converted into organic trace elements to supply the growth of bacteria and promote the growth of mycelium. In addition, selenium (Se) is a mineral with anti-cancer and anti-oxidation functions; zinc (Zn) has minerals that have antioxidant and increased ability to function; bismuth (Ge) is resistant to oxidation and decontamination; chromium (Cr) loses weight, Blood sugar lowering, etc., in the growth formula of the bacteria, adding a trace element compound having a specific effect to the human body, or an inorganic trace element or a yeast powder containing a trace element, which is converted into an organic trace element after being eaten by the bacteria, and the product after fermentation, The human body has more functions and even therapeutic effects.

Oyster shell, Lgneous Rock, Capra Hi reus L, Coral (Coral Hum Japonic touch Ki shinouye), keel (0s Draconi s Coloratus), Dragon tooth (Dens Draconi s), Sea pumice (Purai) Ce), Calcium Carbonate, Calcium Phosphate, Activated Carbon, these minerals are ground into powder and mixed in solid medium (less than 5% by weight) to increase water absorption and avoid sticking of the medium. The medium is breathable and replenishes minerals.

Water keeps the solid medium in a humid state, providing an environment suitable for the growth of bacteria. The moisture content and relative humidity are revised according to different strains and different culture environments.

The addition of trehalose (GsikOu) increases the moisturizing effect of the medium and the environmental resistance of the fermented strain. In addition, in the medium used for liquid fermentation, the carbon source used in the past is mainly sucrose, glucose or molasses. In recent years, starch-based medium has been intensively involved, and as a carbon source, deep fermentation of Ganoderma lucidum is carried out. Its biomass, crude polysaccharide and crude ginseng acid content are high.

In the case of other nitrogen sources, vitamins, minerals and other nutrients, some formulations of solid-state culture can be used, but the dosage and concentration should be additionally corrected. The high concentration of carbon source (high osmotic pressure) has an inhibitory effect on the growth of the cells; the concentration of nitrogen source has little effect on the fermentation cycle, but affects the yield of polysaccharides and secondary metabolites of Ganoderma lucidum.

The solid medium used in the culture dish or test tube is mainly agar or broccoli, and other nutrients are added.

Please refer to FIG. 3, which is an implementation method of the present invention, which includes:

a. pre-treating the medium, including the medium storage tank nitrogen source, carbon source, desired protein, vitamins, minerals and other additives;

b. cooking and sterilizing the medium: then cooling;

C. Select the strain to be produced, culture the culture dish, and enlarge the culture after the flask;

d. Enlarge the strain solution to the liquid fermentation tank (if the inoculum is small, it can be cultured in a triangular flask) Forming a seed inoculating solution;

Solid seeding can also be used as a seed source for inoculation;

e. Inoculation with a vaccination machine;

f. cleaning and sterilizing the culture vessel;

g. The inoculated medium is loaded into the vessel using a medium automatic loader;

h. transport the culture vessel with an automatic access machine or conveyor belt;

1. Enter the automated storage system of the sterile room;

j. Sterilize the sterile room (before the operation);

k. Carry out growth observation and sampling test at any time;

1. If the growth is abnormal, carry out necessary treatment measures;

m. Harvesting the normally grown fermentation product in an automatic access machine or conveyor belt (or other handling machinery, manpower);

n. Drying and grinding;

0. Processing and packaging;

p. sent to the finished product storage area for storage;

q. The finished product can be sampled and analyzed and compared after drying and grinding;

r. Record the optimum ingredient content and optimal culture conditions.

The steps of d, e, f, g can also be changed as follows:

d. Enlarge the strain liquid to the liquid fermentation tank culture (if the inoculum is small, it can be cultured in a triangular flask or other small container) to form the inoculum;

e. Cleaning and sterilizing the culture vessel;

f. The medium is loaded into the vessel using a medium automatic loader;

g. Inoculate with a vaccinator.

Referring to FIG. 4, the present invention adopts a high-pressure culture dish for direct fermentation. The difference from FIG. 3 is that the medium is directly cooked and sterilized in a high-pressure culture vessel in step bl, and the sterilizing cooking vessel is a fermentation culture vessel, which saves the general The step of cleaning and sterilizing the culture vessel; therefore, the step e can be directly inoculated in the culture vessel, and the step of separately injecting the inoculated base into another fermentation vessel is omitted.

The above content is represented by an automated process. In the implementation, warehouse-based fermentation is the main core. Depending on the species and subjective and objective differences, the links of the processes can be completed in a full-motion, semi-automatic or manual manner; The automatic storage system of the storage rack, the storage of the material (fermentation tank), must be taken out or stored by means of automatic access handling equipment; mobile or rotary shelves (warehousing) The automatic storage system of the rack can be moved or removed by moving or rotating the storage unit of the material (fermentation tank) to a specified position, and the material or the machine directly stores or takes out the material; or the access operation is performed through the automatic access handling device. .

The warehouse-based fermentation method is the most efficient use of environmental management to achieve space, which can be enlarged or reduced according to actual needs.

The detailed description above is a detailed description of a possible embodiment of the present invention, which is not intended to limit the scope of the present invention, and equivalent implementations or modifications that are not departing from the spirit of the invention should be included in the present invention. in.

Industrial applicability

The invention utilizes modern automatic storage technology, cooperates with the design of the sterile room space, and automatically implants the culture medium into the fermentation container and completes the sterilization process, and then implants the bacteria, and the automatic access machine (or conveyor belt, other conveying machinery) The fermentation vessels are placed on the shelves of the storage area, and each shelf is equipped with a growth monitoring device, a stirring device, a vibration device, a separate air or oxygen supply device, and provides all the bacteria needed in the fermentation process. The method and system of the present invention are suitable for industrial applications.

Claims

Rights request

A warehouse tissue culture or microbial fermentation method, comprising:

a. Tissue culture or microbial fermentation in a solid, semi-solid, liquid process in the storage space;

b. managing tissue culture or microbial fermentation processes using automated warehousing methods;

c Monitor environmental conditions such as temperature and humidity at any time;

d. Observe the growth situation at any time and conduct sampling tests to maintain product quality;

e. Output the finished product.

2. A warehousing tissue culture or microbial fermentation process according to claim 1 wherein the process is automated to avoid human exposure.

3. A warehousing tissue culture or microbial fermentation process according to claim 1 wherein the sterility is maintained in a sterile chamber space management and sterile air treatment.

4. A warehouse-type tissue culture or microbial fermentation system, comprising:

- Warehousing space, which can be divided into different working areas, for breeding, medium retort sterilization, cooling, inoculation, filling, fermentation, etc.;

a storage system for material storage/removal, material handling and transportation, medium preparation, inoculation, filling, capping, etc. in different working areas of the storage space; a shelf, placed in the storage space, its setting a plurality of cells to provide a space for tissue culture or microbial fermentation; the cells can be extended in a strip to form a tunneling manner; a sterile air system that maintains air entering the storage space in a sterile state;

A fermenter is placed in a storage cell and subjected to rotational agitation or vibration or rest to achieve the aeration and quality transfer functions required for growth to culture or ferment the strain.

5. The warehouse-type tissue culture or microbial fermentation system according to claim 4, wherein the breeding inoculum is planned to have a breeding area, and the breeding area is provided with a strain fermenting tank, which may be a liquid strain tank or a solid strain tank. It is used to culture the inoculum or solid inoculated pellets. When the number of colonies reaches the set standard, the inoculum is inoculated into the general fermentation tank medium, or the medium is inoculated first, and then filled into the fermentation tank.

6. The warehouse-type tissue culture or microbial fermentation system according to claim 4, wherein the storage space is provided with various sterilization equipment such as high temperature and high pressure sterilization, plasma sterilization, ultraviolet sterilization, etc., to achieve a storage space environment. Requirements for sterilization and sterile space.

7. A warehousing tissue culture or microbial fermentation system according to claim 4, wherein the bottom of the storage compartment is provided with a load bearing platform for providing an accommodation space for other functional devices.

8. The warehouse tissue culture or microbial fermentation system according to claim 4, wherein the storage compartment is provided with a helium gas line and a joint, so that the fermentation tanks of each storage compartment are connected to each other to deliver sterile air into the tank for supply. Use of bacteria.

9. A warehouse tissue culture or microbial fermentation system according to claim 4, wherein the storage compartment is provided with a vibrating device or a rotary agitating device.

10. The warehouse tissue culture or microbial fermentation system according to claim 4, wherein the storage compartment or the storage space is provided with monitoring devices such as temperature, humidity, pH, oxygen content, and light.

11. The warehouse-type tissue culture or microbial fermentation system according to claim 4, wherein the fermentation tank is provided with a stirring blade, the stirring blade is provided with a strong magnet, and a powerful magnet driven by a motor is also arranged under the storage compartment. When the motor rotates, the stirring blade in the fermentation tank can be driven to rotate without bearing.

12. The warehouse tissue culture or microbial fermentation system according to claim 4, wherein the fermentation tank is provided with a venting heat dissipating partition for dividing the medium into a plurality of areas in an appropriate amount to increase ventilation and heat dissipation.

13. A warehouse tissue culture or microbial fermentation system according to claim 4, wherein the shelf is fixed or mobile or rotary to provide space for the fermentation tank to optimize space efficiency.

14. A warehouse tissue culture or microbial fermentation system according to claim 4 wherein the connection of the devices can be accomplished fully automatically, semi-automatically or manually.