A fermentation primary seed tank inoculation system

Abstract

The utility model provides a kind of fermentation primary seed tank inoculation system, including the seed liquid pretreatment module for the sterile transfer of seed liquid cultivated by shaking table to industrial environment, the positive pressure inoculation module for maintaining positive pressure sterile environment and realizing inoculation, the intelligent control module for real-time monitoring and automatically adjusting seed liquid pretreatment module and positive pressure inoculation module;Seed liquid pretreatment module and positive pressure inoculation module are separately connected, and seed liquid pretreatment module, positive pressure inoculation module are electrically connected with intelligent control module.The utility model provides a kind of fermentation primary seed tank inoculation system, by setting seed liquid pretreatment module and positive pressure inoculation module, the technical problem that primary seed tank pressure may drop zero in the process of inoculation in the flame ring burning sterilization inoculation mode is solved, prevent the outside pollution from entering due to primary seed tank pressure drop zero in the process of inoculation, reduce the risk of seed liquid contamination in the process of inoculation.

Description

Technical Field

[0001] This utility model relates to the field of industrial fermentation technology, specifically to a primary seed tank inoculation system for fermentation. Background Technology

[0002] Seed culture is an important part of industrial fermentation. It is a preliminary step in microbial fermentation or cell culture. Its purpose is to obtain highly active, highly pure, and sufficient seed liquid by gradually expanding the culture of preserved strains or cells, which is then used to inoculate fermenters or bioreactors for large-scale production.

[0003] A primary seed tank is a small to medium-sized bioreactor used in the seed culture process for the initial amplification of laboratory strains. Its function is to provide highly active seed liquid for subsequent large-scale fermentation. The primary seed tank inoculation system is a specialized device system used in the seed culture process to safely and aseptically transfer the seed liquid cultured in the laboratory to the primary seed tank.

[0004] During the seed culture stage, the rich nutrient components and relatively low microbial biomass of the seed culture medium result in a high risk of contamination. If contamination is not detected in time and the contaminating bacteria are carried into subsequent fermenters, it may cause serious production accidents and irreparable losses. Therefore, strict aseptic control must be implemented in the seed culture process to ensure the safety and stability of the entire fermentation production process.

[0005] As a core component of the seed culture stage, the primary seed tank inoculation system also requires strict aseptic control. Current technology typically employs flame ring sterilization for inoculation. However, this method requires first depressurizing the primary seed tank to zero, then unscrewing the inoculation cap, and pouring the cultured seed solution from the shake flask into the primary seed tank under flame protection. Furthermore, flame ring sterilization has the following drawbacks:

[0006] (1) During inoculation, since the seed liquid is poured directly into the primary seed tank by the staff through the inoculation tube (inoculation port), it is necessary to perform a zeroing operation on the primary seed tank before pouring the seed liquid into the primary seed tank. This causes the positive pressure state originally maintained inside the primary seed tank to be broken, and the pressure balance between the inside of the primary seed tank and the external environment will be broken. This may lead to unfiltered air (containing a large number of bacteria) entering the primary seed tank, increasing the risk of contamination of the primary seed tank.

[0007] (2) There are many uncontrollable factors in the process of using a flame ring for protective inoculation. For example, the flame may be too small to provide sufficient and effective protection. If there are any errors in the operation, such as failure to tighten the inoculation cap in time after inoculating the seed liquid, or the flame going out due to excessive inoculation time, these will all increase the risk of bacterial contamination during the inoculation process.

[0008] Therefore, this solution proposes a primary seed tank inoculation system for fermentation to solve the aforementioned technical problems. Utility Model Content

[0009] The purpose of this invention is to provide a primary seed tank inoculation system for fermentation. By using steam to sterilize the inoculation tube and installing a valve between the inoculation tube and the primary seed tank, a positive pressure environment can be maintained during the inoculation process. This solves the technical problem that the pressure in the primary seed tank may drop to zero during the inoculation process in the flame ring sterilization inoculation method. It prevents external contamination from entering the inoculation process due to the pressure drop in the primary seed tank, and reduces the risk of contamination of the seed liquid during the inoculation process.

[0010] To achieve the above objectives, this utility model proposes a primary seed tank inoculation system for fermentation, comprising a seed liquid pretreatment module for aseptically transferring seed liquid cultured on a shaker to an industrial environment, a positive pressure inoculation module for maintaining a positive pressure aseptic environment and realizing inoculation, and an intelligent control module for real-time monitoring and automatic adjustment of the seed liquid pretreatment module and the positive pressure inoculation module.

[0011] The seed liquid pretreatment module is separately connected to the positive pressure inoculation module, and both the seed liquid pretreatment module and the positive pressure inoculation module are electrically connected to the intelligent control module.

[0012] Furthermore, the seed liquid pretreatment module includes a sterile feeding bottle, a silicone hose 1 detachably connected to the air inlet of the sterile feeding bottle at one end, an air filter detachably connected to the other end of the silicone hose 1, a silicone hose 2 detachably connected to the discharge port of the sterile feeding bottle at one end, and a sterile sealing structure fixedly connected to the other end of the silicone hose 2. The middle section of the silicone hose 2 passes through a peristaltic pump.

[0013] Furthermore, the aseptic sealing structure includes gauze wrapped around the two ends of the silicone tube, kraft paper wrapped around the outside of the gauze, and cotton rope wrapped around the outside of the kraft paper.

[0014] Furthermore, the peristaltic pump includes a controller, a motor electrically connected to the controller, and a pump head driven by the motor;

[0015] The pump head includes a U-shaped pump casing, a rotor connected to the motor drive, and rollers rotatably connected to the outside of the rotor;

[0016] The middle section of the second silicone hose passes between the pump housing and the roller.

[0017] Furthermore, the positive pressure inoculation module includes a primary seed tank in an industrial environment, a pipe connected to the primary seed tank at one end, a tee fixedly connected to the other end of the pipe at one end, an inoculation tube fixedly connected to the other end of the tee at one end, an inoculation cap sealing and covering the other end of the inoculation tube, and a pressure sensor for monitoring the gas pressure state inside the primary seed tank.

[0018] The pressure sensor is located near the inlet of the primary seed tank.

[0019] Furthermore, a tank valve is fixedly connected to one port of the tee near the first pipe, an inoculation valve is fixedly connected to the second port of the tee near the inoculation tube, and a steam valve is fixedly connected to the third port of the tee.

[0020] Furthermore, the tank valve, the inoculation valve, and the steam valve are all electrically connected to the intelligent control module.

[0021] Technical details not described in detail in this solution are based on the conventional understanding and operation of those skilled in the art and will not be elaborated further here.

[0022] The beneficial effects of this utility model are as follows:

[0023] (1) On the one hand, this solution allows the middle section of the silicone tube to pass through the pump head of the peristaltic pump, and uses the peristaltic pump to transport the seed liquid to the inside of the primary seed tank, so that the primary seed tank can complete the inoculation operation under positive pressure without reducing the pressure of the primary seed tank to 0. On the other hand, a three-way valve is set between the primary seed tank and the inoculation tube, and a tank valve, an inoculation valve, and a steam valve are set at the three ports of the three-way valve. Before sterilizing the inoculation tube, the tank valve is closed first, and then the steam valve is opened to sterilize the inoculation tube with steam, thus maintaining the positive pressure environment of the primary seed tank during the sterilization process. That is, this solution maintains the positive pressure environment of the primary seed tank during both the inoculation and sterilization processes, avoids the risk of the pressure of the primary seed tank dropping to zero, prevents the intrusion of external contamination, and reduces the risk of contamination of the seed liquid.

[0024] (2) This method uses steam instead of flame ring burning to sterilize the inoculation tube, making the sterilization intensity and form more controllable and avoiding the problem of insufficient sterilization or flame extinguishing caused by uncontrollable flame size during flame ring burning.

[0025] (3) This solution sets a sterile sealing structure at the port of the silicone tube 2 that is far away from the sterile feeding bottle, so that the seed liquid does not need to come into contact with the outside air during the transfer of the seed liquid from the laboratory to the industrial environment, thereby reducing the risk of contamination of the seed liquid during the transfer from the laboratory to the industrial environment.

[0026] (4) This scheme connects and fixes the silicone tubing to the port of the inoculation tube after sterilization of the inoculation tube, and uses a peristaltic pump to pump the seed liquid from the sterile feeding bottle into the primary seed tank, thus avoiding contact between the seed liquid and the outside air and reducing the risk of contamination of the seed liquid during the inoculation process.

[0027] Turn on the peristaltic pump 16. After it stabilizes, fully open the inoculation valve 242. Under the action of the peristaltic pump 16, all the seed solution in the sterile feeding bottle 11 will be pumped into the primary seed tank 21. After completion, close the tank valve 241. Attached Figure Description

[0028] Figure 1 This is a schematic diagram of the structure of the primary seed tank inoculation system of this utility model.

[0029] Figure 2 This is a schematic diagram of the peristaltic pump of this utility model.

[0030] Figure 3 This is a schematic diagram of the aseptic sealing structure of this utility model.

[0031] The attached diagram is labeled as follows: 1. Seed liquid pretreatment module; 11. Aseptic feeding bottle; 12. Silicone tubing one; 13. Air filter; 14. Silicone tubing two; 15. Aseptic sealing structure; 151. Gauze; 152. Kraft paper; 153. Cotton rope; 16. Peristaltic pump; 161. Pump head; 1611. Pump housing; 1612. Rotor; 1613. Roller; 2. Positive pressure inoculation module; 21. Primary seed tank; 22. Pipe one; 23. Inoculation tube; 24. T-connector; 241. Tank valve; 242. Inoculation valve; 243. Steam valve; 25. Inoculation cap; 3. Intelligent control module. Detailed Implementation

[0032] To more clearly illustrate the technical features of this solution, the following detailed implementation method will be used to explain the solution.

[0033] like Figures 1 to 3 As shown, a primary seed tank inoculation system for fermentation includes a seed liquid pretreatment module 1 for aseptically transferring seed liquid cultured on a shaker to an industrial environment, a positive pressure inoculation module 2 for maintaining a positive pressure aseptic environment and realizing inoculation, and an intelligent control module for real-time monitoring and automatic adjustment of the seed liquid pretreatment module 1 and the positive pressure inoculation module 2.

[0034] The seed liquid pretreatment module 1 is separately connected to the positive pressure inoculation module 2, and both the seed liquid pretreatment module 1 and the positive pressure inoculation module 2 are electrically connected to the intelligent control module.

[0035] The seed culture pretreatment module 1 is used to transfer seed culture from a laboratory environment to an industrial environment in a sterile environment. It provides a safe and sterile environment for the transfer of seed culture through sterile filtration by air filter 13 and sealing protection by sterile sealing structure 15.

[0036] The positive pressure inoculation module 2 is used to maintain a positive pressure sterile environment to achieve seed liquid inoculation. The positive pressure environment can prevent the invasion of external microorganisms and ensure the sterility of the inoculation process.

[0037] Furthermore, the seed liquid pretreatment module 1 includes a sterile feeding bottle 11, a silicone hose 12 detachably connected to the air inlet of the sterile feeding bottle 11 at one end, an air filter 13 detachably connected to the other end of the silicone hose 12, a silicone hose 2 detachably connected to the outlet of the sterile feeding bottle 11 at one end, and a sterile sealing structure 15 fixedly connected to the other end of the silicone hose 2 14. The middle section of the silicone hose 2 14 passes through a peristaltic pump 16.

[0038] The aseptic feeding bottle 11 is made of high-temperature and corrosion-resistant stainless steel and is used to store seed culture in shaker culture.

[0039] Air filter 13 is used to filter the air entering the sterile feeding bottle 11 to prevent external microorganisms from contaminating the seed liquid.

[0040] The silicone tubing 12 is used to connect the air filter 13 to the air inlet of the sterile feeding bottle 11, allowing air filtered by the air filter 13 to enter the sterile feeding bottle 11. The silicone tubing 12 and the air filter 13 are detachably connected, which facilitates the disassembly, cleaning, disinfection and replacement of the silicone tubing 12 and the air filter 13 to ensure the sterility of this inoculation system.

[0041] One end of the silicone tubing 14 is detachably connected to the outlet of the sterile feeding bottle 11, and the other end is fixedly connected to the sterile sealing structure 15, which can ensure the sealing of the seed liquid during transfer.

[0042] The middle section of the silicone tubing 14 passes through the pump head of the peristaltic pump 16. The peristaltic pump 16 delivers the seed liquid by squeezing the silicone tubing 14, which avoids direct contact between the seed liquid and the peristaltic pump 16 and reduces the risk of seed liquid contamination.

[0043] The aseptic sealing structure 15 includes multiple layers of sealing material to enhance the sterility of the seed culture when it is transferred from a laboratory environment to an industrial environment.

[0044] The peristaltic pump 16 is electrically connected to the intelligent control module. The intelligent control module can control the speed and operating status of the peristaltic pump 16 through electrical signals, thereby achieving precise control of the seed liquid delivery speed.

[0045] Furthermore, the aseptic sealing structure 15 includes gauze 151 wrapped around the port of the silicone tube 14, kraft paper 152 wrapped around the outside of the gauze 151, and cotton rope 153 wrapped around the outside of the kraft paper 152.

[0046] The aseptic sealing structure 15 includes gauze 151, kraft paper 152, and cotton rope 153.

[0047] Gauze 151 is the first sealing material of the sterile sealing structure 15, wrapped around the port of the silicone tube 14. Gauze 151 has good air permeability and water absorption, which can initially filter dust and impurities in the air, and at the same time absorb moisture in the seed liquid to prevent leakage of the seed liquid during transportation.

[0048] Kraft paper 152 is the second sealing material of the aseptic sealing structure 15. It is wrapped around the outside of gauze 151 and has good toughness and water resistance. It can further prevent the intrusion of external moisture and microorganisms, and at the same time, it also plays a certain role in protecting gauze 151.

[0049] The cotton rope 153 is the third layer of fixing material for the aseptic sealing structure 15. It is wrapped around the outside of the kraft paper 152 and is used to wrap the gauze 151 and the kraft paper 152 around the port of the silicone tube 14 to form a strong sealing structure, preventing the seed liquid from leaking during the transfer process and reducing the risk of contamination of the seed liquid during the transfer process.

[0050] Furthermore, the peristaltic pump 16 includes a controller, a motor electrically connected to the controller, and a pump head 161 driven by the motor;

[0051] The pump head 161 includes a U-shaped pump housing 1611, a rotor 1612 connected to a motor drive, and a roller 1613 rotatably connected to the outside of the rotor 1612;

[0052] The middle section of the silicone hose 14 passes between the pump housing 1611 and the roller 1613.

[0053] The U-shaped pump housing 1611 is made of high-strength engineering plastic and, together with the roller 1613, forms a closed space to accommodate the silicone hose 14 and complete the extrusion process.

[0054] The rotor 1612 is fixedly connected to the output shaft of the motor. When the peristaltic pump 16 is running, the motor drives the rotor 1612 to rotate, which in turn drives the roller 1613 to rotate around the rotor 1612.

[0055] There are two rollers 1613, which are mainly used to squeeze the silicone tube 2 14 and push the seed liquid inside the silicone tube 2 14 forward.

[0056] The peristaltic pump 16 is existing technology and will not be described in detail here.

[0057] Furthermore, the positive pressure inoculation module 2 includes a primary seed tank 21 set in an industrial environment, a pipe 22 connected to the primary seed tank 21 at one end, a tee 24 fixedly connected to the other end of the pipe 22 at one end, an inoculation tube 23 fixedly connected to the other end of the tee 24, an inoculation cap 25 sealing and covering the port of the inoculation tube 23, and a pressure sensor for monitoring the gas pressure state inside the primary seed tank 21.

[0058] The pressure sensor is located near the inlet of the primary seed tank 21.

[0059] The positive pressure inoculation module 2 is installed in an industrial environment, and the primary seed tank 21 is its core component.

[0060] The primary seed container 21 is made of stainless steel, which is resistant to high temperatures and corrosion to ensure its stability and durability in industrial environments.

[0061] The primary seed tank 21 is equipped with a stirring device to stir the seed liquid and make the seed liquid evenly mixed inside the primary seed tank 21.

[0062] A pressure sensor is installed inside the primary seed tank 21 near the filling port to monitor the tank pressure of the primary seed tank 21 in real time and transmit the real-time tank pressure of the primary seed tank 21 to the intelligent control module.

[0063] The primary seed tank 21 has an air inlet at the bottom and an air outlet at the top. An automatic control valve is installed at the air outlet. The air inlet is fixedly connected to a sterile air preparation system, which continuously supplies sterile air to the primary seed tank 21. The automatic control valve is electrically connected to an intelligent control module. The intelligent control module adjusts the opening of the automatic control valve according to the real-time tank pressure value transmitted by the pressure sensor to control the outflow of air from the primary seed tank 21, thereby maintaining a positive pressure environment in the primary seed tank 21.

[0064] The sterile air preparation system is existing technology and will not be described in detail here.

[0065] Furthermore, a can valve 241 is fixedly connected to one port of the tee 24 near the first pipe 22, an inoculation valve 242 is fixedly connected to the second port of the tee 24 near the second port of the inoculation tube 23, and a steam valve 243 is fixedly connected to the third port of the tee 24.

[0066] Furthermore, the tank valve 241, inoculation valve 242, and steam valve 243 are all electrically connected to the intelligent control module.

[0067] The valve 241 is used to control the passage between the primary seed tank 21 and the three-way valve 24. It is mainly used to isolate the primary seed tank 21 and maintain its positive pressure when sterilizing the inoculation tube 23 by closing the valve 241. That is, by closing the valve 241, the positive pressure environment of the primary seed tank 21 is guaranteed while using steam to sterilize the inoculation tube 23.

[0068] Steam valve 243 is connected to the steam supply pipeline of the boiler upstream and is used to control the steam supply. When steam valve 243 is opened, steam is used to sterilize and purge the inoculation tube 23. The boiler is existing technology and will not be described in detail here.

[0069] Inoculation valve 242 is used to control the inoculation tube passage, regulate the flow rate of seed liquid, and open or close the inoculation channel.

[0070] The specific working process of this utility model:

[0071] For the seed liquid pretreatment module, connect one end of silicone tubing 12 to the air inlet of sterile feeding bottle 11, connect the other end of silicone tubing 12 to air filter 13, connect one end of silicone tubing 14 to the outlet of sterile feeding bottle 2, wrap the other end of silicone tubing 14 with gauze 151 and kraft paper 152 in sequence, and tie it with cotton rope 153. After the assembled silicone tubing 12, silicone tubing 14, sterile feeding bottle 2 and air filter 13 are put into the sterilization cabinet for thorough sterilization, they are transferred to the ultra-clean workbench.

[0072] In the clean bench, the seed culture completed by the shaker is transferred to the sterile feeding bottle 2. After receiving the inoculation notification, the assembled silicone tubing 12, silicone tubing 24, sterile feeding bottle 2, and air filter 13 are transferred to the industrial environment, and silicone tubing 24 is connected and fixed to the peristaltic pump 16.

[0073] In an industrial environment, for the positive pressure inoculation module 2, close the inoculation tank valve 241, open the steam valve 243 and the inoculation valve 242 in sequence, loosen the inoculation cap 25, use steam to fully sterilize the inoculation tube 23, close the steam valve 243, open the tank valve 241, ventilate and dry the inoculation tube 23, tighten the inoculation cap 25 after completion, and wait for inoculation.

[0074] After receiving the vaccination notification, use a spray bottle to spray disinfectant on the primary seed tank 21 and its surrounding environment to sterilize it, and control the pressure inside the primary seed tank 21 at 0.05MPa to ensure a sterile environment.

[0075] Close the inoculation valve 242, unscrew the inoculation cap 25, and sterilize the inoculation tube 23 again with an alcohol swab to ensure a sterile state;

[0076] The inoculation personnel remove the cotton rope 153, kraft paper 152 and gauze 151 from one end of the silicone tubing 214, and connect and fix the silicone tubing 214 to the port of the inoculation tube 23.

[0077] The inoculation personnel turn on the peristaltic pump 16. After the peristaltic pump 16 is running stably, they open the inoculation valve 242 and the tank valve 241. Under the action of the peristaltic pump 16, all the seed liquid in the sterile feeding bottle 11 is pumped into the primary seed tank 21. Then the tank valve 241 is closed.

[0078] Disconnect the silicone tubing 14 from the inoculation tube 23, open the steam valve 243, and use steam to purge and sterilize the inoculation tube 23.

[0079] Close the inoculation valve 242 and steam valve 243, tighten the inoculation cap 25, and complete the entire inoculation process.

[0080] The above are merely preferred embodiments of this utility model and do not constitute any limitation on this utility model. Any equivalent substitutions or modifications made by those skilled in the art to the technical solutions and contents disclosed in this utility model without departing from the scope of the technical solutions of this utility model shall still fall within the protection scope of this utility model.

Claims

1. A primary seed tank inoculation system for fermentation, characterized in that, It includes a seed liquid pretreatment module (1) for aseptically transferring seed liquid cultured in a shaker to an industrial environment, a positive pressure inoculation module (2) for maintaining a positive pressure aseptic environment and realizing inoculation, and an intelligent control module for real-time monitoring and automatic adjustment of the seed liquid pretreatment module (1) and the positive pressure inoculation module (2). The seed liquid pretreatment module (1) is separately connected to the positive pressure inoculation module (2), and both the seed liquid pretreatment module (1) and the positive pressure inoculation module (2) are electrically connected to the intelligent control module.

2. The fermentation primary seed tank inoculation system according to claim 1, characterized in that, The seed liquid pretreatment module (1) includes a sterile feeding bottle (11), a silicone hose 1 (12) detachably connected to the air inlet of the sterile feeding bottle (11) at one end, an air filter (13) detachably connected to the other end of the silicone hose 1 (12), a silicone hose 2 (14) detachably connected to the outlet of the sterile feeding bottle (11) at one end, and a sterile sealing structure (15) fixedly connected to the other end of the silicone hose 2 (14). The middle section of the silicone hose 2 (14) passes through a peristaltic pump (16).

3. The inoculation system for a primary seed tank in fermentation according to claim 2, characterized in that, The sterile sealing structure (15) includes gauze (151) wrapped around the port of the silicone tube (14), kraft paper (152) wrapped around the outside of the gauze (151), and cotton rope (153) wrapped around the outside of the kraft paper (152).

4. The inoculation system for a primary seed tank in fermentation according to claim 2, characterized in that, The peristaltic pump (16) includes a controller, a motor electrically connected to the controller, and a pump head (161) driven by the motor. The pump head (161) includes a U-shaped pump housing (1611), a rotor (1612) connected to the motor drive, and a roller (1613) rotatably connected to the outside of the rotor (1612). The middle section of the second silicone hose (14) passes between the pump housing (1611) and the roller (1613).

5. The inoculation system for a primary seed tank in fermentation according to claim 1, characterized in that, The positive pressure inoculation module (2) includes a primary seed tank (21) set in an industrial environment, a pipe (22) connected to the primary seed tank (21) at one end, a tee (24) fixedly connected to the other end of the pipe (22) at one end, an inoculation tube (23) fixedly connected to the other end of the tee (24) at one end, an inoculation cap (25) sealing and covering the other end of the inoculation tube (23), and a pressure sensor for monitoring the gas pressure state inside the primary seed tank (21). The pressure sensor is located near the inlet of the primary seed tank (21).

6. The inoculation system for a primary seed tank in fermentation according to claim 5, characterized in that, A can valve (241) is fixedly connected to one port of the tee (24) near the first pipe (22), an inoculation valve (242) is fixedly connected to the second port of the tee (24) near the second port of the inoculation tube (23), and a steam valve (243) is fixedly connected to the third port of the tee (24).

7. The fermentation primary seed tank inoculation system according to claim 6, characterized in that, The tank valve (241), the inoculation valve (242), and the steam valve (243) are all electrically connected to the intelligent control module.

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