Continuous biological fermentation reactor

Abstract

The invention relates to the technical field of biological fermentation equipment, in particular to a continuous biological fermentation reactor. The invention aims to solve the technical problems of high energy consumption and low equipment utilization rate of the existing intermittent fermentation. According to the technical scheme, a plurality of isolators are arranged in a horizontal tank body, and the interior of the horizontal tank body is divided into a seed culture section, a production section and an aging section which are sequentially connected in series; sequential flowing of materials in the same functional section is achieved through the in-section communicating mechanism, and controllable isolation and one-way transfer between the functional sections are achieved through the inter-section material transfer mechanism; and the reaction process of a specific stage is enhanced through the in-stage reflux mechanism, and particularly, mature bacterial liquid at the tail end of the strain culture section is refluxed to the starting end so as to realize internal continuous regeneration of the strain. Through systematic innovation of structure and operation, a multi-stage fermentation process is integrated in single equipment, and long-period continuous steady-state production is realized, so that the number of times of empty can sterilization is remarkably reduced, the energy consumption is greatly reduced, and the comprehensive utilization rate of system facilities is greatly improved.

Description

Technical Field

[0001] This invention relates to the field of bio-fermentation equipment technology, specifically to a continuous bio-fermentation reactor. Background Technology

[0002] Bio-fermentation, as the cornerstone of the modern biotechnology industry, is widely used in many key fields such as pharmaceuticals, food, energy, and environmental protection. Currently, industrial-scale bio-fermentation production mainly relies on intermittent or batch-feed operation modes. Although this mode is technologically mature and easy to control, its inherent discontinuity brings many profound challenges and has become a key bottleneck restricting the industry's development towards higher efficiency, lower costs, and greener environmental protection.

[0003] In traditional batch fermentation, a complete production cycle is strictly divided into independent non-production and production phases. The non-production phase includes a series of tedious auxiliary operations such as reactor emptying, CIP (clean-in-place), SIP (sterilization-in-place), cooling, culture medium transfer, and inoculation. Among these, high-temperature, high-pressure steam sterilization of the empty tank is essential to ensure absolute sterility. This process not only consumes enormous amounts of water, electricity, and steam but also generates significant amounts of wastewater. Statistics show that in many fermentation plants, non-production time can account for 30% to 50% of total equipment operating time. This means that expensive fermentation equipment and its associated utilities are idle or operating inefficiently for nearly half the time, significantly reducing asset utilization and overall production efficiency.

[0004] More importantly, the cell growth and product synthesis process in batch mode is a dynamic and non-steady-state process. The cells must go through a lag phase, a logarithmic phase, a stationary phase, and a death phase, resulting in significant fluctuations in production efficiency within batches. To overcome this problem, academia and industry have been exploring continuous culture technology, whose theoretical advantage lies in maintaining the fermentation process in a highly efficient steady state, thereby achieving stable production efficiency and product quality. However, directly scaling up laboratory-scale continuous culture technologies (such as chemostats) to industrial production faces enormous challenges, such as the high susceptibility to phage contamination and strain degradation, and the difficulty in simulating the multi-stage amplification and production process of a "seed tank-fermenter" in a single large-scale reactor.

[0005] Therefore, there is an urgent need in this field for a new technical solution that can break through the limitations of traditional intermittent production mode, realize the continuous production process to significantly reduce energy consumption and improve equipment utilization, and cleverly avoid the instability risks of direct continuous cultivation in large-scale production, creatively constructing a stable, efficient and controllable continuous fermentation microenvironment within a single device. Summary of the Invention

[0006] In view of the shortcomings of the prior art, the purpose of this invention is to provide a continuous bio-fermentation reactor. By dividing the interior of the horizontal tank into a cultivation section, a production section and an aging section, and setting a connecting mechanism within the section and an isolation transfer mechanism between the sections, this invention solves the problem that traditional fermentation requires multiple independent tanks to complete a multi-stage process. It realizes the simulation and optimization of the entire continuous production process from strain amplification to product generation and post-ripening within a single device.

[0007] This invention is achieved using the following technical solution: The continuous bio-fermentation reactor includes: A horizontal tank; Multiple isolators disposed inside the horizontal tank body, the isolators dividing the interior of the tank body into multiple sequentially connected compartments along its length; The plurality of compartments are functionally divided into at least one culture section for strain amplification and one production section for the generation of the target product, the production section being located downstream of the culture section. The intra-segment connecting mechanism is used to enable the free flow of materials between adjacent compartments within the same functional segment (cultivation segment or production segment); An inter-section material transfer mechanism is set between the cultivation section and the production section to controllably transfer materials from the end of the cultivation section to the front of the production section, and to form an effective isolation between the cultivation section and the production section to prevent disorderly flow of materials. The intra-segment reflux mechanism is used to reflux the material from the downstream compartment of a functional segment back to the upstream compartment of the same functional segment in order to enhance the reaction process within that functional segment.

[0008] This invention integrates the "seed expansion culture" and "main fermentation production" processes, which traditionally require multiple independent tanks (such as seed tanks, primary seed tanks, and fermentation tanks), into a single horizontal tank. Through a combination of internal functional partitioning, interconnection within partitions, material transfer isolation between partitions, and enhanced material circulation within partitions, it simulates and optimizes the continuous culture process, fundamentally changing the intermittent production model.

[0009] The aforementioned intra-segment communication mechanism is a material flow channel located at the bottom of adjacent isolators within the same functional segment. This design is simple in structure and utilizes gravity or a small pressure difference to allow the fermentation broth to flow sequentially between the various compartments within the same functional segment (such as the production segment), forming a piston-flow effect and ensuring the gradual conversion of materials.

[0010] The inter-segment transfer mechanism is a differential transfer device, and the isolator equipped with this device does not have a material flow channel at its bottom. The inlet of this device is located at a specific liquid level in the final chamber of the upstream functional segment. Only when the liquid level in that chamber reaches the preset height can the liquid be transferred to the downstream functional segment through this device. Simultaneously, the bottom of the isolator equipped with the differential transfer device is sealed, without any material flow channel. This achieves effective isolation between functional segments, preventing disorderly back-mixing of downstream high-concentration metabolites or microorganisms at different growth stages into the upstream culture segment, ensuring the independence and stability of the process conditions in each functional segment.

[0011] The aforementioned intra-section reflux mechanism is an isobaric liquid reflux device. Its inlet is located in the upper-middle liquid phase zone of the separator, and its outlet is located in the top gas phase zone of the target reflux chamber. This reflux device typically utilizes the gas lifting principle to return the liquid material from the upper part of the separator (usually the area with the strongest bacterial activity) to the upstream separator in the same functional section through a pipe located in the gas phase space of the reactor. Because the reflux pipe is above the liquid surface, uncontrollable cross-contamination caused by the siphon effect is avoided, achieving a gentle and controllable circulation of specific materials without disrupting the system pressure balance, thus enhancing the reaction.

[0012] The multiple compartments also include an aging section connected downstream of the production section; and an inter-section transfer mechanism is also provided between the production section and the aging section. The aging section is used for further transformation, modification, or maturation of the products in the later stages of fermentation, which is particularly important for the fermentation of certain antibiotics, organic acids, etc. The production section and the aging section are also connected by an inter-section transfer mechanism to achieve controllable material transfer and inter-section isolation.

[0013] The material outlet height of the inter-segment transfer mechanism between the cultivation section and the production section is higher than that of the inter-segment transfer mechanism between the production section and the aging section. A discharge port is located at the end of the aging section, and the height of this discharge port is lower than the material outlet height of the inter-segment transfer mechanism between the production section and the aging section, thus forming a cascaded gravity flow. This physical structure ensures that the material can only flow unidirectionally from the cultivation section → production section → aging section → discharge port, effectively preventing unorganized backflow from the later section to the earlier section.

[0014] The in-segment reflux mechanism includes a first reflux device installed in the cultivation section and a second reflux device installed in the production section; The first reflux device is used to reflux the mature bacterial solution in the last stage of the culture section back to the starting stage of the culture section. The second reflux device is used to reflux the bacterial solution from the final stage of the production section to a separate compartment in the middle of the production section.

[0015] Specifically, the first reflux unit returns the highly concentrated and vigorous mature bacterial solution from the final chamber of the culture stage to the starting chamber of the culture stage. This is equivalent to establishing an internal, continuous bacterial inoculation system, enabling the starting chamber to continuously obtain high-quality "new inoculum," greatly enhancing the vigor of the bacterial community, reducing the frequency and dependence on external fresh bacterial inoculation, and thus lowering the risk of contamination caused by frequent inoculation.

[0016] The second reflux unit returns the liquid from the final stage of the production section to a separate compartment in the middle of the production section. This accelerates the distribution of high-density, high-activity microorganisms throughout the entire production section, balancing the reaction and stabilizing production intensity. Simultaneously, it ensures a more stable concentration of the liquid delivered to the aging section, which is beneficial for controlling the aging process and ensuring uniformity in the final product quality.

[0017] The bottom plate of the horizontal tank has a downward slope along the direction of material flow from the cultivation section to the production section. Specifically, to facilitate smooth material flow, the bottom plate of the horizontal tank can be designed with a downward slope of 0 to 30 degrees along the direction of material flow from the cultivation section to the production and aging sections. This gentle slope helps the fermentation broth and microorganisms overcome flow resistance under gravity, smoothly passing through the material channels within the sections and preventing material from accumulating at the bottom of the tank and forming dead zones.

[0018] The isolator is a cooling isolator with an internal cooling channel. The cooling medium enters the isolator through a pipe to efficiently exchange heat with the fermentation broth. Since each compartment is in contact with the cooling isolator, precise and differentiated temperature control can be achieved for different fermentation stages (breeding, production, aging).

[0019] The final chamber of the cultivation, production, and / or aging sections is equipped with a double-layered, sealed manhole, forming a sealed space that can be filled with antibacterial liquid. This design constitutes a reliable sterile barrier, minimizing the intrusion of external microorganisms even during maintenance or observation. Simultaneously, the equipped level gauge, which does not contact the material, ensures real-time monitoring without compromising the system's airtightness.

[0020] Compared with the prior art, the beneficial effects of the present invention are: (1) By dividing the interior of the horizontal tank into a cultivation section, a production section and an aging section, and setting up a connecting mechanism within the section and an isolation transfer mechanism between the sections, the structural design solves the problem that traditional fermentation requires multiple independent tanks (seed tank, fermentation tank and post-processing tank) to complete the multi-stage process. It realizes the simulation and optimization of the entire continuous production process from strain amplification to product generation and then to post-ripening in a single device.

[0021] (2) By setting up an in-segment reflux mechanism in the cultivation section to return the bacterial solution from the mature inoculation chamber to the starting chamber, continuous internal inoculation and self-renewal and enhancement of the bacterial strain are realized, significantly reducing the frequency of inoculation of fresh external bacterial strains, thereby reducing the risk of contamination caused by frequent inoculation operations, and enhancing the long-term production vitality and stability of the entire microbial community.

[0022] (3) By transforming intermittent batch production into long-cycle continuous steady-state production, the problem of high energy consumption and long non-production time caused by frequent empty tank sterilization in the traditional intermittent production mode is solved. This invention can extend the continuous production cycle of a single tank by two to ten times, thereby significantly reducing the number of sterilizations, significantly reducing the consumption of steam, water and electricity, and raising the overall utilization rate of equipment to a new level, bringing significant economic and environmental benefits.

[0023] (4) By setting up a second reflux mechanism in the production section to return the bacterial liquid from the final stage to the middle section, and by forming a cascaded gravity flow between sections through the differential transfer device, the concentration, cell distribution and flow of the fermentation liquid in each functional section are effectively controlled, ensuring the stability of the liquid concentration delivered to the later section, which is conducive to the stability of the aging process and the uniformity of the final product quality, and facilitates subsequent separation and purification operations. Attached Figure Description

[0024] Figure 1 This is a schematic diagram of the continuous bio-fermentation reactor of the present invention; Figure 2 For the present invention Figure 1 Enlarged view of point A in the middle; In the diagram: 1. Isobaric liquid reflux device in the cultivation section; 2. Shell of the bio-fermentation reactor; 3. Exhaust port; 4. Double-layer manhole in the cultivation section; 5. Level gauge in the cultivation section; 6. Differential transfer device from the cultivation section to the production section; 7. Primary isobaric liquid reflux device; 8. Production chamber; 9. Double-layer manhole in the production section; 10. Secondary isobaric liquid reflux device; 11. Level gauge in the production section; 12. Aging chamber; 13. Differential transfer device from the production section to the aging section; 14. Double-layer manhole in the aging section; 15. Level gauge in the aging section; 16. Pressure gauge; 17. Steam inlet; 18. 19. Sterile air inlet; 20. Sterile water inlet; 21. Feed inlet A; 22. Feed inlet B; 23. Transplanting port; 24. External cooling jacket semi-circular tube; 25. pH meter; 26. Isolator; 27. Dissolved oxygen meter; 28. Thermometer; 29. ​​Inoculation tube; 30. Cooling water connection A; 31. Cooling water connection B; 32. Inoculation chamber; 33. Cultivation section final stage chamber; 34. Production section material passage; 35. Production section final stage chamber; 36. Aging section material passage; 37. Aging section final stage chamber; 38. Continuous discharge port; 39. Manhole. Detailed Implementation

[0025] To make the objectives and technical solutions of this invention clearer, the invention will be further described in detail below.

[0026] To enable those skilled in the art to better understand the present invention, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments. Obviously, the described embodiments are merely some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0027] like Figure 1-2 As shown, Figure 1 This is a schematic diagram of a continuous bio-fermentation reactor provided in an embodiment of the present invention.

[0028] In one specific embodiment of this application, the continuous bio-fermentation reactor includes an integral bio-fermentation reactor shell 2 that can withstand an internal pressure of 0.3 MPa. Inside the bio-fermentation reactor shell 2, a plurality of vertical isolators 25 are provided, which divide the interior of the bio-fermentation reactor shell 2 into n sequentially connected compartments along the length of the bio-fermentation reactor shell 2.

[0029] Understandably, these n compartments are functionally divided into three coherent areas according to the bio-fermentation process: the cultivation section at one end of the reactor, which includes inoculation compartment 31 to the final cultivation compartment 32 (mature seed compartment); the production section that follows, which includes multiple production compartments 8 up to the final production compartment 34; and the aging section at the other end of the reactor, which includes multiple aging compartments 12 up to the final aging compartment 36.

[0030] Within the same functional section (e.g., production section or aging section), the bottom of adjacent isolators 25 is provided with a production section material flow channel 33 and an aging section material flow channel 35. These material flow channels constitute an intra-section communication mechanism, allowing the fermentation broth to flow sequentially between the various compartments of the same functional section under the action of gravity and a small pressure difference, thereby realizing the gradual transformation of materials.

[0031] At the boundaries between different functional sections, such as between the cultivation section and the production section, and between the production section and the aging section, the bottom of the isolator 25 does not have a material passage. Instead, it is equipped with a material transfer device 6 for the difference between the cultivation section and the production section, and a material transfer device 13 for the difference between the production section and the aging section. The material transfer device 6 for the difference between the cultivation section and the production section is used to transfer the mature bacterial liquid from the last chamber 32 of the cultivation section into the production section, and the material transfer device 13 for the difference between the production section and the aging section is used to transfer the fermentation liquid from the last chamber 34 of the production section into the aging section. This structure achieves effective isolation between functional sections. Only when the liquid level in the upstream functional section reaches a certain height can the liquid cross the weir and enter the downstream functional section, thereby preventing disorderly back-mixing of materials.

[0032] To ensure unidirectional and orderly material flow, the outlet height of the material transfer device 6 between the cultivation and production sections is higher than the outlet height of the material transfer device 13 between the production and aging sections. Simultaneously, the continuous discharge port 37 located on the final stage silo 36 of the aging section is lower than the outlet height of the material transfer device 13. This stepped height design creates a cascaded gravity flow within the entire reactor, physically ensuring that the material can only flow unidirectionally from the cultivation section → production section → aging section.

[0033] In addition, to enhance the reaction process and maintain bacterial activity, the reactor is equipped with an isobaric liquid reflux unit 1 in the culture section, a primary isobaric liquid reflux unit 7, and a secondary isobaric liquid reflux unit 10. The isobaric liquid reflux unit 1 in the culture section returns the high-concentration mature bacterial solution from the final stage chamber 32 of the culture section to the initial stage chamber 31 of the culture section, serving as a new source of inoculum for continuous internal inoculation. The secondary isobaric liquid reflux unit 10 returns the bacterial solution from the final stage chamber 34 of the production section to a production chamber 8 in the middle of the production section, accelerating the distribution of highly active bacteria from the later stages to the earlier stages and balancing the reaction rate throughout the production section. The pipes of these reflux units are all located in the gas phase space above the liquid surface of the reactor, operating on the principle of gas lifting to achieve controllable intra-stage circulation.

[0034] In terms of temperature control, the outer shell 2 of the bio-fermentation reactor is equipped with a cooling jacket consisting of an external cooling jacket semi-circular pipe 23, and the isolator 25 itself is also designed as a cooling isolator with an internal cooling channel. Cooling water is introduced through cooling water inlet A 29 and cooling water inlet B 30, which can efficiently exchange heat with the fermentation broth.

[0035] To ensure long-term aseptic operation and effective monitoring of the system, each functional section's final compartment is equipped with double-layered sealed manholes (double-layered manhole 4 in the culture section, double-layered manhole 9 in the production section, and double-layered manhole 14 in the aging section), non-contact level gauges (level gauge 5 in the culture section, level gauge 11 in the production section, and level gauge 15 in the aging section), and manhole 38. Simultaneously, each compartment is equipped with necessary process interfaces and monitoring instruments as needed, including an exhaust port 3, a pressure gauge 16, a steam inlet 17, a sterile air inlet 18, a sterile water inlet 19, feed inlets (feed inlet A 20, feed inlet B 21), a transfer port 22, a pH meter 24, a dissolved oxygen meter 26, a thermometer 27, and an inoculum tube 28, etc., to achieve comprehensive online monitoring and control of the fermentation process.

[0036] In summary, this invention constructs an integrated continuous production system by dividing the interior of the bio-fermentation reactor shell 2 into a cultivation section, a production section, and an aging section, and by using a material flow channel to achieve intra-section connectivity, a differential material transfer device to achieve inter-section isolation and material transfer, and an isobaric liquid reflux device to achieve enhanced intra-section circulation. This effectively extends the production cycle, reduces energy consumption, and improves equipment utilization.

[0037] The above-mentioned continuous bio-fermentation reactor, when in operation, includes the following steps: (1) Preparation stage: In accordance with the empty tank disinfection procedure, thorough sterilization is carried out through the reactor steam inlet 17. After sterilization is qualified, the tank is kept at a positive pressure of 0.03 MPa through the sterile air inlet 18 to prevent external contamination. Sterile process water is injected through the sterile water inlet 19 of the water-continuous disinfection system to the set level.

[0038] (2) Cultivation Stage: Start the culture medium continuous sterilization system and inject the culture medium into each compartment of the cultivation section through the inlet. Then, inoculate the culture section with the rated amount of original seed solution through the transfer port 22 or the inoculation tube 28. Start the isobaric liquid reflux device 1 in the cultivation section to return the liquid in the mature seed compartment (the last compartment 32 of the cultivation section) to the inoculation compartment 31, forming an internal circulation to enhance the amplification and activation of the strain. During this stage, monitor parameters such as cell density and morphology in real time.

[0039] (3) Transition to Production Stage: Once the inoculum in the incubation stage reaches the quality standards required by the process, the liquid level will rise and cross the weir of the differential transfer device by adjusting the pressure of the incubation stage and the production stage (making the pressure of the incubation stage slightly higher than that of the production stage). This will continuously transfer the mature inoculum to the first compartment of the production stage, marking the system's entry into continuous production. At the same time, fresh reaction raw materials are continuously added to the production stage, and the isobaric liquid reflux device in the production stage is activated to return part of the inoculum in the final compartment 34 of the production stage to the middle of the production stage, in order to maintain the high reaction efficiency and stability of the entire production stage.

[0040] (4) Transition to aging and discharge stage: The liquid material in the production section flows downstream sequentially through the bottom material channel in the silo. When the reaction liquid in the final stage silo 34 of the production section reaches the set parameters, the pressure is adjusted (making the pressure in the production section slightly higher than that in the aging section by about 0.01 MPa), and the liquid material enters the aging section through the differential transfer device. The liquid material continues to flow slowly through the material channel in the aging section to complete the final maturation process. When the liquid material indicators in the final stage silo 36 of the aging section are qualified, the continuous discharge port 37 can be opened to continuously output the finished fermentation liquid for subsequent separation and purification processes.

Claims

1. A continuous bio-fermentation reactor, characterized in that, include A horizontal tank; Multiple isolators disposed inside the horizontal tank body, the isolators dividing the interior of the tank body into multiple sequentially connected compartments along its length; The plurality of compartments are functionally divided into at least one culture section for strain amplification and one production section for the generation of the target product, the production section being located downstream of the culture section. The intra-segment connecting mechanism is used to enable the free flow of materials between adjacent compartments within the same functional segment; An inter-section material transfer mechanism is set between the cultivation section and the production section to controllably transfer materials from the end of the cultivation section to the front of the production section, and to form an effective isolation between the cultivation section and the production section to prevent disorderly flow of materials. The intra-segment reflux mechanism is used to reflux the material from the downstream compartment of a functional segment back to the upstream compartment of the same functional segment in order to enhance the reaction process within that functional segment.

2. The continuous bio-fermentation reactor according to claim 1, characterized in that, The intra-segment communication mechanism is a material passage located at the bottom of adjacent isolators within the same functional segment.

3. The continuous bio-fermentation reactor according to claim 1, characterized in that, The inter-segment material transfer mechanism is a differential material transfer device, and the bottom of the isolator equipped with the differential material transfer device does not have a material passage.

4. The continuous biofermentation reactor according to claim 1, characterized in that, The aforementioned intra-segment reflux mechanism is an isobaric liquid reflux device, with its feed inlet located in the upper-middle liquid phase zone of the partition chamber and its discharge outlet located in the top gas phase zone of the target reflux chamber.

5. The continuous biofermentation reactor according to claim 1, characterized in that, The plurality of compartments also includes an aging section connected downstream of the production section; and an inter-section transfer mechanism is also provided between the production section and the aging section.

6. The continuous bio-fermentation reactor according to claim 5, characterized in that, The material outlet height of the inter-segment transfer mechanism between the cultivation section and the production section is higher than the material outlet height of the inter-segment transfer mechanism between the production section and the aging section; the end of the aging section is provided with a discharge port, the height of which is lower than the material outlet height of the inter-segment transfer mechanism between the production section and the aging section, thereby forming a cascaded gravity flow.

7. The continuous biofermentation reactor according to claim 1, characterized in that, The in-segment reflux mechanism includes a first reflux device installed in the cultivation section and a second reflux device installed in the production section; The first reflux device is used to reflux the mature bacterial solution in the last stage of the culture section back to the starting stage of the culture section. The second reflux device is used to reflux the bacterial solution from the final stage of the production section to a separate compartment in the middle of the production section.

8. The continuous biofermentation reactor according to claim 1, characterized in that, The bottom plate of the horizontal tank has a downward slope along the direction in which the material flows from the cultivation section to the production section.

9. The continuous biofermentation reactor according to claim 1, characterized in that, The isolator is a cooling isolator with an internal cooling channel.

10. The continuous biofermentation reactor according to claim 5, characterized in that, The final chamber of the cultivation section, production section and / or aging section is provided with a double-layered sealed manhole, and a sealed space that can be filled with antibacterial liquid is formed between the inner and outer layers of the manhole.

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