Microbial fermentation process automatic on-line detection and feed automatic feedback control system

Abstract

The present application relates to the technical fields of microbial fermentation control, and provides an automatic online detection and automatic feedback control system for a microbial fermentation process, a ceramic membrane is arranged in a filtering cavity, the ceramic membrane separates the filtering cavity into an inner cavity and an outer cavity, the inner cavity is connected with a fermentation tank through a material discharge pipeline and a cell backflow pipeline respectively, the outer cavity is connected with a dilution mixing pool through a clear liquid discharge pipeline, a dilution water supply pipeline is connected with the dilution mixing pool, and the dilution mixing pool is connected with a sampling pool; a pressure sensor and a pneumatic diaphragm regulating valve are arranged on the cell backflow pipeline in sequence along the cell flow direction. By arranging the ceramic membrane in the filtering cavity, the cells in the fermentation liquid can be backflowed to the fermentation tank, the clear liquid of the fermentation liquid can flow to the dilution mixing pool, and the diluted clear liquid can be detected and analyzed in the sampling pool, so that sampling can be completed online without contaminating the fermentation liquid in the fermentation tank, and the whole fermentation process is kept free of contamination risk.

Description

Technical Field

[0001] This invention relates to the field of microbial fermentation control technology, and in particular to an automatic online detection and automatic feeding feedback control system for microbial fermentation processes. Background Technology

[0002] With the development of bioengineering, bio-fermentation technology has also been widely developed in agriculture, industry, medicine, pharmacology, energy, environmental protection, and chemical raw materials. During microbial fermentation, the fermentation broth needs to be tested to ensure that no contaminating microorganisms are generated and that the desired microorganisms or metabolites are obtained. During sampling, the culture medium sample is manually removed from the culture tank. Before sampling, high-pressure steam is introduced through the sampling port for 30 minutes for sterilization. Then, the sampling port is opened to release the first portion of the sample, while the middle and later portions are quickly sealed. After sampling, high-pressure steam is introduced through the sampling port for 30 minutes for sterilization. The removed sample is transferred to a centrifuge for centrifugation, and the supernatant is transferred to analytical equipment for analysis. Generally, the centrifuge and analytical equipment are located in other laboratories, with unavoidable spatial and temporal gaps. After obtaining the sample test results, the operator determines whether to replenish the feed based on calculations. If replenishment is needed, the feed replenishment device is manually controlled to quantitatively replenish the feed. The existing technologies described above have the following technical drawbacks: First, the sterilization operations before and after sampling are cumbersome and time-consuming, which cannot meet the needs of high sampling frequency. Second, the entire testing process includes manual sampling, centrifugation, testing, and replenishment, which takes a long time. The content of the tested components in the culture tank is constantly changing. By the time the replenishment amount is calculated, the content of the tested components in the culture tank may have changed, thus preventing the replenishment from achieving the expected results. Third, the amount of manual sampling cannot be precisely controlled, making it impossible to achieve multiple sampling and monitoring in the cell culture industry where the amount of culture medium is small. Fourth, the entire existing testing process requires manual operation, which consumes a lot of labor costs. Summary of the Invention

[0003] This invention provides an automatic online detection and automatic feeding feedback control system for microbial fermentation processes, which solves the problem of cumbersome manual sampling procedures in the prior art.

[0004] This invention provides an automatic online detection and automatic feeding feedback control system for a microbial fermentation process, comprising: a fermenter, a filter chamber, a dilution mixing tank, a sampling tank, and a dilution water supply pipeline; The filtration chamber is equipped with a ceramic membrane, which divides the filtration chamber into an inner chamber and an outer chamber. The inner chamber is connected to the fermenter through a material discharge pipeline and a cell return pipeline, respectively. The outer chamber is connected to the dilution mixing tank through a clear liquid discharge pipeline. The dilution water supply pipeline is connected to the dilution mixing tank, and the dilution mixing tank is connected to the sampling tank. A pressure sensor and a pneumatic diaphragm regulating valve are sequentially installed along the flow direction of the bacteria in the bacterial return pipeline. The opening of the pneumatic diaphragm regulating valve is adjusted according to the pressure value of the pressure sensor. The automatic online detection and automatic feeding feedback control system for microbial fermentation process provided by the present invention further includes a stirring device, which includes a stir bar disposed in the dilution mixing tank and a magnetic stirrer disposed outside the dilution mixing tank. The magnetic stirrer is configured to drive the stir bar to rotate relative to the dilution mixing tank.

[0005] According to the present invention, an automatic online detection and automatic feeding feedback control system for microbial fermentation process further includes a steam supply pipeline, which is connected to the cell return pipeline via a pneumatic three-way diaphragm valve. The cell return pipeline is connected to the fermenter via a first aseptic sampling valve, which is also connected to a sampling return and wastewater discharge pipeline. A first temperature sensor, a first pneumatic diaphragm valve, and a first manual diaphragm valve are sequentially arranged on the sampling return and wastewater discharge pipeline. The material discharge pipeline is connected to the fermenter via a second aseptic sampling valve, which is also connected to a sampling discharge and wastewater discharge pipeline. A second temperature sensor, a second pneumatic diaphragm valve, and a second manual diaphragm valve are sequentially arranged on the sampling discharge and wastewater discharge pipeline. During the sterilization of the filter chamber, the first sterile sampling valve and the second sterile sampling valve are in the closed state, while the pneumatic three-way diaphragm valve, the pneumatic diaphragm regulating valve, the first pneumatic diaphragm valve, the first manual diaphragm valve, the second pneumatic diaphragm valve, and the second manual diaphragm valve are all in the open state, and the temperature of the steam supplied to the steam supply pipeline is controlled by the interlocking of the first temperature sensor and the second temperature sensor.

[0006] According to the present invention, an automatic online detection and automatic feeding feedback control system for microbial fermentation process further includes a cleaning liquid supply pipeline, wherein the cleaning liquid supply pipeline is connected to the steam supply pipeline via a first pneumatic three-way ball valve, and the cleaning liquid supply pipeline is also connected to the external cavity via a pneumatic ball valve; a second pneumatic three-way ball valve is provided on the clear liquid discharge pipeline; wherein a first pump body is provided on the material discharge pipeline; The first and second sterile sampling valves are closed, the pneumatic three-way diaphragm valve is opened, the first pneumatic three-way ball valve is switched to connect to the cleaning fluid supply pipeline, the first pump body reverses, and the cleaning water is discharged from the sampling return sewage pipeline and the sampling discharge sewage pipeline; after the bacterial return pipeline is cleaned, the first and second pneumatic diaphragm valves are closed, the first pump body rotates forward, and the pneumatic ball valve is opened.

[0007] According to the present invention, an automatic online detection and automatic feeding feedback control system for microbial fermentation process is provided, wherein a third pneumatic three-way ball valve is provided on the pipeline connecting the pneumatic ball valve to the outer cavity, and the third pneumatic three-way ball valve is connected to the outer cavity venting pipeline.

[0008] The automatic online detection and automatic feeding feedback control system for microbial fermentation process provided by the present invention further includes a parameter detector, which is installed in the clear liquid discharge pipeline and is used to detect at least one of sugar content and light transmittance. A second pump body is provided in the dilution water supply pipeline.

[0009] According to the present invention, an automatic online detection and automatic feeding feedback control system for microbial fermentation process is provided, wherein the sampling tank is connected to a waste liquid collection tank through an outlet pipeline.

[0010] An automatic online detection and automatic feeding feedback control system for a microbial fermentation process provided by the present invention further includes: a control motherboard, a sampling component, a feeding pipeline, and a detection component, wherein the feeding pipeline is connected to the fermenter, and the sampling component, the feeding pipeline, and the detection component are all electrically connected to the control motherboard; The control board is configured to control the operation of the sampling component to draw liquid from the sampling pool, move the liquid to the detection component for detection, and control the feeding pipeline to feed the fermenter according to the detection result of the detection component.

[0011] According to the present invention, an automatic online detection and automatic feeding feedback control system for microbial fermentation process is provided. The sampling component includes a moving mechanism and a sampling needle. The sampling needle is connected to the moving mechanism, and the moving mechanism is electrically connected to the control motherboard. Under the control of the control motherboard, the moving mechanism drives the sampling needle to draw liquid from the sampling pool and moves the liquid to the detection component for detection.

[0012] According to the present invention, an automatic online detection and automatic feeding feedback control system for microbial fermentation process is provided. The feeding pipeline includes a first feeding pipeline and / or a second feeding pipeline. The first feeding pipeline is sequentially provided with a third pneumatic diaphragm valve, a first flow meter, a first pneumatic regulating valve, and a fourth pneumatic diaphragm valve. The second feeding pipeline is sequentially provided with a fifth pneumatic diaphragm valve, a second flow meter, a second pneumatic regulating valve, and a sixth pneumatic diaphragm valve. The first flow meter, the first pneumatic regulating valve, the second flow meter, and the second pneumatic regulating valve are all electrically connected to the control main board. The control board is configured to control the first pneumatic regulating valve and / or the second pneumatic regulating valve to operate according to the detection result of the detection component, so that the flow value of the first flow meter and / or the flow value of the second flow meter are both equal to the corresponding target flow value.

[0013] The automatic online detection and automatic feeding feedback control system for microbial fermentation provided by this invention allows the microbial cells in the fermentation broth to flow back into the fermenter by setting a ceramic membrane in the filtration chamber, while the clear liquid of the fermentation broth flows to the dilution and mixing tank. The diluted clear liquid can be detected and analyzed in the sampling tank. In other words, online sampling can be completed without contaminating the fermentation broth in the fermenter, keeping the entire fermentation process free from the risk of contamination. In addition, the entire sampling process is simple to operate and saves time and effort. Attached Figure Description

[0014] To more clearly illustrate the technical solutions in this invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0015] Figure 1 This is a schematic diagram of the automatic online detection and automatic feeding feedback control system for the microbial fermentation process provided by the present invention.

[0016] Figure label: 1. Fermentation tank; 2. Material discharge pipeline; 21. Second aseptic sampling valve; 22. Second pneumatic diaphragm valve; 23. Second manual diaphragm valve; 24. Second temperature sensor; 25. First pump body; 3. Ceramic membrane; 31. Inner cavity; 32. Outer cavity; 4. Bacterial reflux pipeline; 41. Pneumatic three-way diaphragm valve; 42. Pressure sensor; 43. Pneumatic diaphragm regulating valve; 44. First aseptic sampling valve; 45. First pneumatic diaphragm valve; 46. First manual diaphragm valve; 47. First temperature sensor; 5. Steam supply pipeline; 51. First pneumatic three-way ball valve; 6. Cleaning fluid supply pipeline; 61. Third pneumatic three-way ball valve; 62. Pneumatic ball valve; 7. External cavity venting pipeline; 8. Clear liquid discharge pipeline; 81. Second pneumatic three-way ball valve; 82. Third pump body; 83. Parameter detector; 9. External drainage pipe; 10. Dilution and mixing tank; 11. Dilution water supply pipeline; 111. Second pump body; 12. Sampling tank; 121. Waste liquid collection tank; 13. Sampling assembly; 131. Moving mechanism; 132. Sampling needle; 14. Detection components; 141. Enzyme membrane electrode detector; 142. Liquid chromatography detector; 15. Control motherboard; 16. First feed line; 161. Third pneumatic diaphragm valve; 162. Seventh pneumatic diaphragm valve; 163. First flow meter; 164. First pneumatic regulating valve; 165. Fourth pneumatic diaphragm valve; 166. Eighth pneumatic diaphragm valve; 17. Second feed line; 171. Fifth pneumatic diaphragm valve; 172. Ninth pneumatic diaphragm valve; 173. Second flow meter; 174. Second pneumatic regulating valve; 175. Sixth pneumatic diaphragm valve; 176. Tenth pneumatic diaphragm valve. Detailed Implementation

[0017] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0018] like Figure 1 As shown, the automatic online detection and automatic feeding feedback control system for microbial fermentation process according to an embodiment of the present invention includes: a fermenter 1, a filter chamber, a dilution mixing tank 10, a sampling tank 12, and a dilution water supply pipeline 11.

[0019] A ceramic membrane 3 is installed in the filtration chamber, dividing the filtration chamber into an inner chamber 31 and an outer chamber 32. The pore size of the ceramic membrane 3 is 50~200nm. The inner chamber 31 is connected to the fermenter 1 through the material discharge pipe 2 and the cell return pipe 4, meaning that the inner chamber 31 can form a loop with the inside of the fermenter 1. The outer chamber 32 is connected to the dilution mixing tank 10 through the clear liquid discharge pipe 8, the dilution water supply pipe 11 is connected to the dilution mixing tank 10, and the dilution mixing tank 10 is connected to the sampling tank 12.

[0020] The ceramic membrane 3 is an asymmetric membrane formed by a special process using inorganic ceramic materials. The wall of the ceramic membrane 3 is densely covered with micropores. Under pressure, the raw liquid flows inside or outside the membrane tube. Small molecules permeate through the membrane, while large molecules are retained by the membrane, thereby achieving separation, concentration, purification, and environmental protection purposes. For example, a large columnar ceramic membrane 3 can be placed in the filtration chamber.

[0021] In this system, after the fermentation broth in fermenter 1 is filtered through the filtration chamber, the bacterial cells remain in the inner cavity 31 and flow back to fermenter 1, while the clarified liquid enters the outer cavity 32 and flows into the dilution mixing tank 10. Additionally, the dilution water supply pipeline 11 provides dilution water to the dilution mixing tank 10 according to a preset dilution ratio, allowing the dilution water to mix with the clarified liquid flowing into the dilution mixing tank 10. The mixed liquid then flows into the sampling tank 12 for subsequent sampling and analysis of its components, and the fermenter 1 is replenished based on the analysis results.

[0022] like Figure 1 As shown, a pressure sensor 42 and a pneumatic diaphragm regulating valve 43 are sequentially installed on the bacterial reflux pipeline 4 along the bacterial flow direction.

[0023] The automatic online detection and automatic feeding feedback control system for the microbial fermentation process can be equipped with a control motherboard 15, and the pressure sensor 42 and the pneumatic diaphragm regulating valve 43 are all electrically connected to the control motherboard 15.

[0024] Specifically, the fermentation broth in fermenter 1 enters the inner cavity 31 of ceramic membrane 3. The liquid pressure in the inner cavity 31 of ceramic membrane 3 is controlled by a pneumatic diaphragm regulating valve 43 and a pressure sensor 42, causing the liquid in the inner cavity 31 to permeate through the tube wall of ceramic membrane 3 at a certain speed to reach the outer cavity 32 of ceramic membrane 3, thus achieving the purpose of cell recovery. For example, the control board 15 controls the opening degree of the pneumatic diaphragm regulating valve 43 based on the pressure value of the pressure sensor 42 and the target pressure value.

[0025] In this embodiment of the invention, by setting a ceramic membrane 3 in the filter chamber, the bacteria in the fermentation broth can be refluxed back into the fermenter 1, while the clear liquid of the fermentation broth flows to the dilution mixing tank 10. The diluted clear liquid can be tested and analyzed in the sampling tank 12. In other words, online sampling can be completed without contaminating the fermentation broth in the fermenter 1, keeping the entire fermentation process free from the risk of contamination. In addition, the entire sampling process is simple to operate and can save time and effort.

[0026] In some embodiments, such as Figure 1 As shown, the automatic online detection and automatic feeding feedback control system for microbial fermentation process also includes a stirring device. The stirring device includes a stir bar installed inside the dilution mixing tank 10 and a magnetic stirrer installed outside the dilution mixing tank 10. The magnetic stirrer is configured to drive the stir bar to rotate relative to the dilution mixing tank.

[0027] Specifically, the stir bar is a magnet, and the magnetic stirrer is used to generate a changing magnetic field to drive the stir bar to rotate. Under the action of the stirring device, the permeate from the ceramic membrane 3 can be mixed evenly with the dilution water. The stirring device can be electrically connected to the control board 15, in other words, the start and stop of the stirring device can be controlled by the control board 15.

[0028] In some embodiments, such as Figure 1 As shown, the automatic online detection and automatic feeding feedback control system for the microbial fermentation process also includes a steam supply pipeline 5. The steam supply pipeline 5 is connected to the cell return pipeline 4 via a pneumatic three-way diaphragm valve 41. The cell return pipeline 4 is connected to the fermenter 1 via a first sterile sampling valve 44. The first sterile sampling valve 44 is also connected to a sampling return and wastewater discharge pipeline. A first temperature sensor 47, a first pneumatic diaphragm valve 45, and a first manual diaphragm valve 46 are sequentially installed on the sampling return and wastewater discharge pipeline. The material discharge pipeline 2 is connected to the fermenter 1 via a second sterile sampling valve 21. The second sterile sampling valve 21 is also connected to a sampling discharge and wastewater discharge pipeline. A second temperature sensor 24, a second pneumatic diaphragm valve 22, and a second manual diaphragm valve 23 are sequentially installed on the sampling discharge and wastewater discharge pipeline. The steam supply pipeline 5 is used to provide high-temperature steam.

[0029] It is understandable that the pneumatic three-way diaphragm valve 41, the first sterile sampling valve 44, the first temperature sensor 47, the first pneumatic diaphragm valve 45, the first manual diaphragm valve 46, the second sterile sampling valve 21, the second temperature sensor 24, the second pneumatic diaphragm valve 22, and the second manual diaphragm valve 23 can all be electrically connected to the control mainboard 15.

[0030] It should be noted that the pneumatic three-way diaphragm valve 41 is located between the pressure sensor 42 and the pneumatic diaphragm regulating valve 43, and the first sterile sampling valve 44 is located downstream of the pneumatic diaphragm regulating valve 43. The first pump body 25 is located between the second sterile sampling valve 21 and the outer cavity 32.

[0031] In this embodiment of the invention, during the sterilization of the filter chamber, the first sterile sampling valve 44 and the second sterile sampling valve 21 are in the closed state, while the pneumatic three-way diaphragm valve 41, the pneumatic diaphragm regulating valve 43, the first pneumatic diaphragm valve 45, the first manual diaphragm valve 46, the second pneumatic diaphragm valve 22, and the second manual diaphragm valve 23 are all in the open state. This allows external steam to enter the ceramic membrane 3 and its associated pipelines for sterilization. The speed and temperature of the steam are controlled by interlocking with the first temperature sensor 47 and the second temperature sensor 24, ensuring that the ceramic membrane 3 and its associated pipelines reach a sterile state. After a set time, the valves are closed, completing the sterilization operation of the ceramic membrane 3 and its associated pipelines. The clear liquid discharge pipeline 8 can be connected to an external cavity drain pipeline 9.

[0032] It is particularly important to note that when normal offline sampling is required, the first manual diaphragm valve 46 or the second manual diaphragm valve 23 should be manually closed, and the second sterile sampling valve 21 and the second pneumatic diaphragm valve 22 should be opened by controlling the main board 15, or the first sterile sampling valve 44 and the first pneumatic diaphragm valve 45 should be opened, and then the first manual diaphragm valve 46 or the second manual diaphragm valve 23 should be manually opened to complete the sampling operation.

[0033] In some embodiments, such as Figure 1 As shown, it also includes a cleaning fluid supply pipeline 6, which is connected to the steam supply pipeline 5 via a first pneumatic three-way ball valve 51. The cleaning fluid supply pipeline 6 is also connected to the outer cavity 32 via a pneumatic ball valve 62. A second pneumatic three-way ball valve 81 is installed on the clear liquid discharge pipeline 8. Additionally, a first pump body 25 is installed on the material discharge pipeline 2, which pumps the fermentation liquid from the fermenter 1 into the inner cavity 31 of the ceramic membrane 3. The first pump body 25, the first pneumatic three-way ball valve 51, the pneumatic ball valve 62, and the second pneumatic three-way ball valve 81 can all be electrically connected to the control main board 15. The cleaning water provided by the cleaning fluid supply pipeline 6 can be divided into sterile water and cleaning water with a certain antibacterial ability, such as dilute acid, dilute alkali, sodium hypochlorite, etc. It should be noted that the cleaning water with antibacterial ability is used for cleaning when the ceramic membrane 3 is clogged and for system protection when the system is shut down.

[0034] Understandably, a cleaning operation can be performed after the entire fermentation process is completed. Specifically, the first aseptic sampling valve 44 and the second aseptic sampling valve 21 are closed, the pneumatic three-way diaphragm valve 41 is opened, the first pneumatic three-way ball valve 51 is switched to connect to the cleaning solution supply pipeline 6, and the first pump body 25 reverses direction. Thus, cleaning water is discharged from the sampling return drain pipeline and the sampling discharge drain pipeline to the external sewage pipeline. The cell return pipeline 4 will be cleaned first, and after cleaning, the first pneumatic diaphragm valve 45 and the second pneumatic diaphragm valve 22 are closed. The first pump body 25 rotates forward, and the pressure sensor 42 controls the pressure in the inner cavity 31 of the ceramic membrane 3. The pneumatic ball valve 62 opens, allowing the outer cavity 32 of the ceramic membrane 3 to be cleaned, leading to the sampling tank 12 and related pipelines. The second pneumatic three-way ball valve 81 is connected to the outer cavity drain pipeline 9.

[0035] In some embodiments, such as Figure 1 As shown, a third pneumatic three-way ball valve 61 is installed on the pipeline connecting the pneumatic ball valve 62 to the outer cavity 32.

[0036] It should be noted that the third pneumatic three-way ball valve 61 can be connected to an external cavity venting pipe 7. In this way, after the external cavity 32 of the ceramic membrane 3 to the sampling pool 12 and related pipes are cleaned, high-temperature gas can be introduced through the external cavity venting pipe 7 so that the entire system can be dried quickly.

[0037] In some embodiments, such as Figure 1 As shown, the automatic online detection and automatic feeding feedback control system for the microbial fermentation process also includes a parameter detector 83, which is installed in the clear liquid discharge pipeline 8. The parameter detector 83 is used to detect at least one of the sugar content and light transmittance. A second pump body 111 is installed on the dilution water supply pipeline 11. The parameter detector 83 is located between the second pneumatic three-way ball valve 81 and the dilution mixing tank 10. A third pump body 82 can also be installed between the second pneumatic three-way ball valve 81 and the parameter detector 83. Thus, the parameter detector 83, the second pump body 111, and the third pump body 82 can all be electrically connected to the control main board 15.

[0038] It should be noted that the parameter detector 83 may include a first instrument for detecting sugar content and a second instrument for detecting light transmittance, or the parameter detector 83 may simultaneously detect sugar content and light transmittance.

[0039] For example, the parameter detector 83 includes a detection cell and an incident light generator and a receiving light detector located on opposite sides of the detection cell. The detection cell is made of quartz or glass. The parameter detector 83 is used to detect the absorbance, transmittance or refractive index of a liquid at different wavelengths.

[0040] It should be noted that when the inner cavity 31 of the ceramic membrane 3 is cleaned by the cleaning water supplied by the cleaning fluid supply pipeline 6, the cleaning can be confirmed as complete when the transmittance of the permeate is close to 100% as detected by the parameter detector 83.

[0041] Understandably, the third pump 82 will pump the permeate to the dilution mixing tank 10. Based on the sugar content measured by the parameter detector 83 and the historical data stored in the control main board 15, the control main board 15 can determine the required dilution ratio. Under the control of the control main board 15, the second pump 111 will pump the corresponding amount of dilution water into the dilution mixing tank 10.

[0042] As shown above, by monitoring the absorbance value at 600nm using parameter detector 83, a sudden increase in absorbance indicates that bacteria have permeated through the ceramic membrane 3 into the outer cavity 32, triggering an alarm and shutdown for maintenance. By detecting the liquid sugar content, a correlation is established between sugar content and product content. Based on the calculated product concentration and the required detection concentration on the control board 15, the dilution factor is calculated, and the corresponding dilution is automatically completed in the dilution mixing tank 10. Furthermore, for repetitive production processes, the dilution factors required for different fermentation cycles are relatively fixed. The control board 15 can execute the system's default dilution factor. When the detection result deviates significantly from the expected result, the dilution factor for the next sample is re-determined based on the deviation, and an alarm is triggered to prompt the main control personnel to investigate the cause.

[0043] In some embodiments, such as Figure 1As shown, the sampling tank 12 is connected to a waste liquid collection tank 121 via an outlet pipe. It should be noted that the waste liquid collection tank 121 typically has sufficient capacity to hold all the waste liquid flowing out of the sampling tank 12. The waste liquid collection tank 121 ensures that the waste liquid flowing out of the sampling tank 12 will not pollute the environment. For example, the liquid passing through the dilution mixing tank 10 enters the bottom of the sampling tank 12 and then flows out from the top side of the sampling tank 12 to the waste liquid collection tank 121.

[0044] Microbial fermentation refers to the process by which microorganisms, under suitable conditions, transform raw materials into products needed by humans through specific metabolic pathways. In recent years, with the continuous advancement and development of biotechnology, bio-fermentation engineering technology and related products are increasingly impacting people's daily lives. In the bio-fermentation process, multi-stage continuous fermentation systems have high fermentation production efficiency, and therefore are usually chosen for fermentation operations. During fermentation, continuous feeding into fermenter 1 is required via a feedstock to maintain the fermentation process.

[0045] Thus, in some embodiments, such as Figure 1 As shown, the automatic online detection and automatic feeding feedback control system for the microbial fermentation process also includes: a control motherboard 15, a sampling component 13, a feeding pipeline, and a detection component 14. The feeding pipeline is connected to the fermenter 1, and the sampling component 13, the feeding pipeline, and the detection component 14 are all electrically connected to the control motherboard 15.

[0046] The control board 15 is configured to control the operation of the sampling component 13 to draw liquid from the sampling pool 12, move the liquid to the detection component 14 for detection, and control the feeding pipeline to feed the fermenter 1 according to the detection result of the detection component 14.

[0047] In this embodiment of the invention, under the control of the control motherboard 15, the fermenter 1 can realize real-time automatic separation of bacteria, automatic dilution, and automatic sampling and detection. Based on the detection results and preset programs, the feeding rate or frequency can be automatically adjusted to realize automatic control of feeding during the fermentation process. In this way, continuous sampling can be achieved, the sampling density can be subdivided to the minute level, and there is no risk of contamination during the entire sampling process.

[0048] In some embodiments, such as Figure 1 As shown, the sampling component 13 includes a moving mechanism 131 and a sampling needle 132. The sampling needle 132 is connected to the moving mechanism 131, and the moving mechanism 131 is electrically connected to the control motherboard 15. Under the control of the control motherboard 15, the moving mechanism 131 drives the sampling needle 132 to draw liquid from the sampling pool 12 and move the liquid to the detection component 14 for detection.

[0049] It should be noted that the moving mechanism 131 can be a two-dimensional moving mechanism 131 or a three-dimensional moving mechanism 131. Both the two-dimensional moving mechanism 131 and the three-dimensional moving mechanism 131 can be robotic arms in the prior art, and no specific limitation is made here.

[0050] The detection component 14 may include an enzyme membrane electrode detector 141 and a liquid chromatography detector 142. The enzyme membrane electrode detector 141 and the liquid chromatography detector 142 are placed on the experimental platform. The moving mechanism 131 can move linearly horizontally and vertically. The sampling cell 12 has a detachable rubber cover on top. The sampling needle 132, driven by the moving mechanism 131, is inserted into the sampling cell 12 and draws a quantitative amount of sample liquid. After sampling, the sampling needle 132 accurately positions the enzyme membrane electrode detector 141 or the liquid chromatography detector 142 for corresponding injection and detection under the action of the moving mechanism 131.

[0051] In some embodiments, such as Figure 1 As shown, the feeding pipeline includes a first feeding pipeline 16 and / or a second feeding pipeline 17. That is, the feeding pipeline includes the first feeding pipeline 16, or the feeding pipeline includes the second feeding pipeline 17, or the feeding pipeline includes the first feeding pipeline 16 and the second feeding pipeline 17.

[0052] The first feed line 16 is equipped with a third pneumatic diaphragm valve 161, a first flow meter 163, a first pneumatic regulating valve 164, and a fourth pneumatic diaphragm valve 165 in sequence. The second feed line 17 is equipped with a fifth pneumatic diaphragm valve 171, a second flow meter 173, a second pneumatic regulating valve 174, and a sixth pneumatic diaphragm valve 175 in sequence. The first flow meter 163, the first pneumatic regulating valve 164, the second flow meter 173, and the second pneumatic regulating valve 174 are all electrically connected to the control main board 15.

[0053] The control board 15 is configured to control the first pneumatic regulating valve 164 and / or the second pneumatic regulating valve 174 to operate according to the detection result of the detection component 14, so that the flow value of the first flow meter 163 and / or the flow value of the second flow meter 173 are both equal to the corresponding target flow value.

[0054] It should be noted that during the fermentation process, the first aseptic sampling valve 44 and the second aseptic sampling valve 21 are open, while the first pneumatic diaphragm valve 45 and the second pneumatic diaphragm valve 22 are closed. The first pump body 25 pumps the fermentation liquid from the fermenter 1 into the inner cavity 31 of the ceramic membrane 3. The liquid pressure in the inner cavity 31 of the ceramic membrane 3 is controlled by the opening of the pneumatic diaphragm regulating valve 43 and the pressure sensor 42, causing the liquid in the inner cavity 31 of the ceramic membrane 3 to pass through the tube wall of the ceramic membrane 3 at a certain speed to reach the outer cavity 32 of the ceramic membrane 3, thus achieving the purpose of filtration and sterilization. At the same time, the third pump body 82 pumps the permeate to the dilution mixing tank 10. Based on the sugar content measured by the parameter detector 83 and the historical data saved on the control mainboard 15, the required dilution ratio is determined. The second pump body 111 then pumps the corresponding amount of dilution water into the dilution mixing tank 10. After being stirred and mixed by the stirring device, the water flows to the sampling tank 12. Driven by the moving mechanism 131, the sampling needle 132 periodically draws liquid from the sampling pool 12 according to a preset program and transfers it to the enzyme membrane electrode detector 141 or the liquid chromatography detector 142 for detection. The detection results are fed back to the control mainboard 15. The control mainboard 15 calculates the flow rate of the first feeding line 16 and / or the second feeding line 17 based on the detection value and preset conditions, and adjusts the opening of the first pneumatic regulating valve 164 and / or the second pneumatic regulating valve 174 so that the flow rate of the first flow meter 163 and / or the second flow meter 173 reaches the new set value.

[0055] It should be noted that the outlet of the third pneumatic diaphragm valve 161 is connected to the seventh pneumatic diaphragm valve 162, and the inlet of the fourth pneumatic diaphragm valve 165 is connected to the eighth pneumatic diaphragm valve 166. The outlet of the fifth pneumatic diaphragm valve 171 is connected to the ninth pneumatic diaphragm valve 172, and the inlet of the sixth pneumatic diaphragm valve 175 is connected to the tenth pneumatic diaphragm valve 176. The seventh, eighth, ninth, and tenth pneumatic diaphragm valves 172 and 176 are all electrically connected to the control mainboard 15.

[0056] Thus, during sterilization of the fermenter (either empty or fully operated), the third pneumatic diaphragm valve 161 is closed, while the fourth, seventh, and eighth pneumatic diaphragm valves 165, 162, and 166 are opened. This allows the high-temperature steam inside the fermenter to be expelled from the system through the seventh and eighth pneumatic diaphragm valves 162 and 166, completing the sterilization of the section between the third pneumatic diaphragm valve 161 and the fermenter 1 in the first feed line 16. The second feed line follows the same principle. The sterilization principle of the pipeline between the third pneumatic diaphragm valve 161 and the feed source is the same. At this time, the fourth pneumatic diaphragm valve 165 is closed, while the third, seventh, and eighth pneumatic diaphragm valves 161, 162, and 166 are open. The sterilization method of the second feed line is the same as that of the first feed line.

[0057] It is particularly important to note that the automatic online detection and automatic feeding feedback control system for microbial fermentation is intended for sampling and detection; therefore, the entire system's piping and chamber volume is very small. Specifically, the total volume of the outer chamber 32 of the ceramic membrane 3, the dilution mixing tank 10, the sampling tank 12, and the connecting pipes is designed to be 10-20 mL. Taking a total volume of 10 mL as an example, without sample dilution, the permeation rate of the ceramic membrane 3 only needs to reach 10 mL / min to ensure that the sample state at the time of sampling in the sampling tank 12 represents the fermentation broth from 1 minute prior. The detection time of the enzyme membrane electrode detector 141 is generally within 10 minutes, and the detection time of the liquid chromatography detector 142 is generally within 30 minutes. Since fermentation is a gradual process, a delay of less than one hour is perfectly acceptable for experimental or production processes.

[0058] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. An automatic online detection and automatic feeding feedback control system for a microbial fermentation process, characterized in that, include: Fermentation tank, filtration chamber, dilution mixing tank, sampling tank, and dilution water supply pipeline; The filtration chamber is equipped with a ceramic membrane, which divides the filtration chamber into an inner chamber and an outer chamber. The inner chamber is connected to the fermenter through a material discharge pipeline and a cell return pipeline, respectively. The outer chamber is connected to the dilution mixing tank through a clear liquid discharge pipeline. The dilution water supply pipeline is connected to the dilution mixing tank, and the dilution mixing tank is connected to the sampling tank. A pressure sensor and a pneumatic diaphragm regulating valve are sequentially installed along the flow direction of the bacteria on the bacterial return pipeline. The opening degree of the pneumatic diaphragm regulating valve is adjusted according to the pressure value of the pressure sensor.

2. The automatic online detection and automatic feeding feedback control system for microbial fermentation process according to claim 1, characterized in that, It also includes a stirring device, which includes a stir bar disposed inside the dilution mixing tank and a magnetic stirrer disposed outside the dilution mixing tank, the magnetic stirrer being configured to drive the stir bar to rotate relative to the dilution mixing tank.

3. The automatic online detection and automatic feeding feedback control system for microbial fermentation process according to claim 1, characterized in that, It also includes a steam supply pipeline, which is connected to the cell return pipeline via a pneumatic three-way diaphragm valve. The cell return pipeline is connected to the fermenter via a first aseptic sampling valve. The first aseptic sampling valve is also connected to a sampling return and sludge discharge pipeline. A first temperature sensor, a first pneumatic diaphragm valve, and a first manual diaphragm valve are sequentially installed on the sampling return and sludge discharge pipeline. The material discharge pipeline is connected to the fermenter via a second aseptic sampling valve. The second aseptic sampling valve is also connected to a sampling discharge and sludge discharge pipeline. A second temperature sensor, a second pneumatic diaphragm valve, and a second manual diaphragm valve are sequentially installed on the sampling discharge and sludge discharge pipeline. During the sterilization of the filter chamber, the first sterile sampling valve and the second sterile sampling valve are in the closed state, while the pneumatic three-way diaphragm valve, the pneumatic diaphragm regulating valve, the first pneumatic diaphragm valve, the first manual diaphragm valve, the second pneumatic diaphragm valve, and the second manual diaphragm valve are all in the open state, and the temperature of the steam supplied to the steam supply pipeline is controlled by the interlocking of the first temperature sensor and the second temperature sensor.

4. The automatic online detection and automatic feeding feedback control system for microbial fermentation process according to claim 3, characterized in that, It also includes a cleaning fluid supply pipeline, which is connected to the steam supply pipeline via a first pneumatic three-way ball valve. The cleaning fluid supply pipeline is also connected to the outer cavity via a pneumatic ball valve. A second pneumatic three-way ball valve is provided on the clear liquid discharge pipeline. A first pump body is provided on the material discharge pipeline. The first and second sterile sampling valves are closed, the pneumatic three-way diaphragm valve is opened, the first pneumatic three-way ball valve is switched to connect to the cleaning fluid supply pipeline, the first pump body reverses, and the cleaning water is discharged from the sampling return sewage pipeline and the sampling discharge sewage pipeline; after the bacterial return pipeline is cleaned, the first and second pneumatic diaphragm valves are closed, the first pump body rotates forward, and the pneumatic ball valve is opened.

5. The automatic online detection and automatic feeding feedback control system for microbial fermentation process according to claim 4, characterized in that, A third pneumatic three-way ball valve is provided on the pipeline connecting the pneumatic ball valve to the outer cavity, and the third pneumatic three-way ball valve is connected to the outer cavity venting pipeline.

6. The automatic online detection and automatic feeding feedback control system for microbial fermentation processes according to any one of claims 1 to 5, characterized in that, It also includes a parameter detector, which is installed in the clear liquid discharge pipeline. The parameter detector is used to detect at least one of the sugar content and light transmittance. A second pump body is installed in the dilution water supply pipeline.

7. The automatic online detection and automatic feeding feedback control system for microbial fermentation processes according to any one of claims 1 to 5, characterized in that, The sampling pool is connected to a waste liquid collection pool via an outlet pipe.

8. The automatic online detection and automatic feeding feedback control system for microbial fermentation processes according to any one of claims 1 to 5, characterized in that, It also includes: a control motherboard, a sampling component, a feeding pipeline, and a detection component, wherein the feeding pipeline is connected to the fermenter, and the sampling component, the feeding pipeline, and the detection component are all electrically connected to the control motherboard; The control board is configured to control the operation of the sampling component to draw liquid from the sampling pool, move the liquid to the detection component for detection, and control the feeding pipeline to feed the fermenter according to the detection result of the detection component.

9. The automatic online detection and automatic feeding feedback control system for microbial fermentation process according to claim 8, characterized in that, The sampling component includes a moving mechanism and a sampling needle. The sampling needle is connected to the moving mechanism, and the moving mechanism is electrically connected to the control motherboard. Under the control of the control motherboard, the moving mechanism drives the sampling needle to draw liquid from the sampling pool and moves the liquid to the detection component for detection.

10. The automatic online detection and automatic feeding feedback control system for microbial fermentation process according to claim 8, characterized in that, The feeding pipeline includes a first feeding pipeline and / or a second feeding pipeline. The first feeding pipeline is sequentially equipped with a third pneumatic diaphragm valve, a first flow meter, a first pneumatic regulating valve, and a fourth pneumatic diaphragm valve. The second feeding pipeline is sequentially equipped with a fifth pneumatic diaphragm valve, a second flow meter, a second pneumatic regulating valve, and a sixth pneumatic diaphragm valve. The first flow meter, the first pneumatic regulating valve, the second flow meter, and the second pneumatic regulating valve are all electrically connected to the control main board. The control board is configured to control the first pneumatic regulating valve and / or the second pneumatic regulating valve to operate according to the detection result of the detection component, so that the flow value of the first flow meter and / or the flow value of the second flow meter are both equal to the corresponding target flow value.

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