Intelligent temperature control stirring reaction kettle for microbial polysaccharide fermentation and fermentation method

Abstract

The application provides a kind of intelligent temperature control stirring reaction kettle for microbial polysaccharide fermentation and fermentation method, it is related to fermentation technical field, including: reaction kettle body, the intermediate position of the reaction kettle body is slidably installed with rotating tube;The outer side of the feeding pipe is provided with multiple openings, the outer side of feeding pipe is slidably installed with sealing pipe, and the side end of feeding pipe is installed with spring, and the side end of feeding pipe is connected with the inner side end of sealing pipe through spring;The outer side of the sealing pipe is provided with multiple openings, and the openings can coincide with the openings on the outer side of the feeding pipe.Nutrient solution is evenly fed into the reaction kettle at different positions through the openings on the feeding pipe and the sealing pipe, and the mixing effect of the fermentation broth and the supplemented nutrient solution is effectively enhanced at different positions, ensuring the quality of microbial polysaccharide produced by fermentation, solving the problem that a large amount of nutrient solution will gather in the upper layer of fermentation broth due to factors such as density, and the mixing effect with the lower fermentation broth is poor.

Description

Technical Field

[0001] This invention relates to the field of fermentation technology, and in particular to an intelligent temperature-controlled stirred reactor and fermentation method for microbial polysaccharide fermentation. Background Technology

[0002] Microbial polysaccharides are polysaccharide substances produced by bacteria and fungi through metabolism. They have good thickening, gelling, and biocompatibility properties and are widely used in food, medicine, cosmetics and other fields. Microbial polysaccharide fermentation is mostly carried out in reaction vessels and subsequent extraction and processing to obtain microbial polysaccharides.

[0003] In existing technologies for microbial polysaccharide fermentation, nutrient replenishment is required as carbon or nitrogen sources are gradually consumed. However, when the replenished nutrient solution is directly added to the fermentation broth in the reactor, a large amount of nutrient solution will accumulate on the upper layer of the fermentation broth due to factors such as density, resulting in poor mixing with the fermentation broth below. If this situation is improved by increasing the stirring rate, an excessively high stirring rate will cause shear damage to the polysaccharide molecules, affecting the stability of the polysaccharide molecular weight. Summary of the Invention

[0004] To address the shortcomings of existing technologies, this invention provides an intelligent temperature-controlled stirred reactor and fermentation method for microbial polysaccharide fermentation, thereby solving the problems mentioned in the background section.

[0005] To solve the above-mentioned technical problems, the present invention is achieved through the following technical solution: This invention relates to an intelligent temperature-controlled stirring reactor and fermentation method for microbial polysaccharide fermentation, specifically comprising: a reactor body, wherein two guide rings are installed on the inner side of the reactor body, and the bottom of the guide rings has an inclined structure; two positioning vertical rods are installed on the bottom of the reactor body, wherein the bottom of the positioning vertical rods is provided with limiting protrusions; a drive cylinder is installed on the outer bottom of the reactor body, and a traction base plate is installed on the output end of the bottom of the drive cylinder; the traction base plate is slidably installed on the outer side of the two positioning vertical rods, and a servo motor is installed on the bottom of the traction base plate, and a gear is installed on the output end of the servo motor; a rotating tube is slidably installed in the middle position of the reactor body; the bottom of the rotating tube... A gear is installed at the bottom of the reactor body, which meshes with a gear at the output end of a servo motor at the bottom of the traction base plate. The bottom of the rotating tube is rotatably mounted on the traction base plate. Multiple evenly distributed delivery pipes are installed on the outer side of the rotating tube. Multiple evenly distributed openings are provided on the outer side of the delivery pipes. A sealing pipe is slidably installed on the outer side of the delivery pipes, and a spring is installed on the side end of the delivery pipes. The side end of the delivery pipes is connected to the inner end of the sealing pipes through the springs. Multiple evenly distributed openings are provided on the outer side of the sealing pipes, and the openings can coincide with the openings on the outer side of the delivery pipes. A guide block is installed on the outer end of the sealing pipes. The guide block is slidably mounted on the inner side of the guide ring.

[0006] Furthermore, the reactor body has a cylindrical structure, with a feed inlet at the top and a display panel installed on the outer side. The sidewalls of the reactor body have a sandwich structure, and a connecting flange is installed on the outer side. Support legs are installed at the bottom edge of the reactor body. An online detection module is installed on the outer side of the rotating tube, which integrates a pH sensor, dissolved oxygen electrode, temperature sensor, viscosity sensor, and turbidity sensor. A feed hood is installed at the top of the rotating tube, penetrating the top of the reactor body.

[0007] Furthermore, a flow guide is rotatably installed on the outside of the feed hood; a nutrient solution storage tank is installed on the outside of the reactor body; a drive pump is installed on the top of the nutrient solution storage tank, and a flow guide pipe is installed on the output end of the drive pump; the side end of the flow guide pipe is connected to the outside of the flow guide hood.

[0008] Furthermore, multiple inner coils are installed on the inner side of the reactor body; a diversion pipe is installed on the outer side of each inner coil; an injection pipe is installed through the side end of each diversion pipe, penetrating the side of the reactor body; a connecting flange is installed on the top of the injection pipe; multiple multi-functional feeding pipes are installed on the top of the reactor body; a stirring blade is installed on the outer side of the rotating pipe; an air inlet pipe is installed in the jacket of the reactor body; a quick connector is installed at the outer end of the air inlet pipe, and multiple conveying pipes are installed on the side of the air inlet pipe; the conveying pipe has a ring structure, and an aeration pipe is provided on the side of the conveying pipe; the aeration pipe is located inside the reactor body.

[0009] Furthermore, a positioning cylinder is installed on one side of the top of the reactor body; a normally closed sealing plate is rotatably installed at the bottom of the positioning cylinder, and a spring is installed at the top of the positioning cylinder, with a push plate installed at the top of the spring; a movable tube is installed at the side end of the push plate.

[0010] Furthermore, nozzles in different directions are installed at the bottom of the movable tube through the top of the positioning cylinder; the bottom of the movable tube and the nozzles are located inside the positioning cylinder; a purification-type drive pump is installed on one side of the top of the reactor body; a water flow pipe is installed on the output end of the top of the purification-type drive pump; the side end of the water flow pipe is connected to the top of the movable tube.

[0011] Furthermore, a method for fermenting microbial polysaccharides includes the following steps: First, clean the reactor body by rinsing the inner wall of the reactor, stirring blades, feeding pipes and sealing pipes with high-pressure water to remove residual impurities. Then, immerse it in sodium hydroxide solution for 30-40 minutes, and then rinse it with deionized water until the pH is neutral. Second, staged sterilization: First, use high temperature of saturated steam to sterilize the inside of the reactor body, then inject fermentation medium into the reactor body, use high temperature sterilization to obtain clean medium, and then cool it down to the fermentation start temperature. Third, strain activation and inoculation: inject the pre-cultured microbial seed liquid into the reaction vessel; Fourth, for the initial parameter settings, the temperature, stirring speed, pH value, and dissolved oxygen value are adjusted according to the type of microorganism. Fifth, phased intelligent temperature control: the fermentation stages are divided into the lag phase, logarithmic phase and stationary phase, and temperature is controlled at different stages. Sixth, by adjusting the rotation speed of the stirring disc at different fermentation stages, combined with the amount of oxygen injected, the dissolved oxygen content and viscosity of the fermentation broth can be adjusted. 7. Nutrient supplementation and regulation: The amount of cells is monitored by the turbidity sensor on the online detection module. When the turbidity reaches 800-1000 NTU, the drive pump on the nutrient solution tank is turned on to add carbon and nitrogen sources to the reactor to prevent the substrate from being depleted and causing fermentation to stop. 8. During the fermentation process monitoring and miscellaneous microbial control stage, the online detection module collects real-time data on pH, dissolved oxygen, temperature, viscosity, and turbidity. The number of microorganisms is detected by injecting air into a multi-functional feed pipe. At the same time, the sealing of the reactor body is checked regularly to reduce the intrusion of miscellaneous microorganisms. 9. By detecting that the polysaccharide concentration, cell volume, and pH value of the fermentation broth are in a stable state, a stable polysaccharide fermentation broth is obtained.

[0012] Furthermore, the segmented sterilization in step two includes: In the first stage, 121°C saturated steam is introduced into the jacket and inner coil of the reactor, maintaining a pressure of 0.12-0.15 MPa for 30-40 minutes to sterilize and kill microorganisms on the surface of the reactor. In the second stage, the prepared fermentation medium is injected into the reactor body, and 115℃ saturated steam is introduced through the inner coil to maintain the pressure at 0.08-0.1MPa for 20-25 minutes to sterilize and avoid high temperature from destroying the nutrients in the medium. In the third stage, after sterilization, cooling is carried out by introducing cooling water through the jacket of the inner wall of the reactor to lower the temperature of the culture medium to the fermentation start temperature. At the same time, sterile air is introduced into the reactor to prevent the invasion of other bacteria. In step three, the preserved microbial strains are inoculated into the seed culture medium and cultured in a shaker for 12-16 hours in advance to the logarithmic phase to obtain the seed liquid. The seed liquid is then injected into the reaction vessel to avoid contamination during the inoculation process. In step four, the temperature is set according to the type of microorganism, for example, xanthan gum fermentation at 28-30℃ and gellan gum fermentation at 30-32℃. The stirring speed is set to 150-200 rpm to ensure that the bacteria are evenly dispersed; The pH value is set according to the type of microorganism, for example, 6.5-7.0 for xanthan gum fermentation and 7.0-7.5 for gellan gum fermentation. It is adjusted by using an alkali or acid addition device through a multi-functional feeding pipe. The dissolved oxygen level is maintained at 20-30% saturation by adjusting the stirring speed and the flow rate of sterile air.

[0013] Furthermore, the segmented temperature control in step five includes: During the delayed period, microbial metabolic heat production is low, so temperature control is mainly achieved through the jacket of the reactor body, with the internal coil providing auxiliary control, to maintain temperature fluctuations ≤ ±0.5℃. During the logarithmic phase, microorganisms multiply rapidly, and metabolic heat production increases sharply. The temperature inside the reactor is collected by the online detection module. When the temperature is 0.3℃ higher than the set value, cold water is injected into the inner coil for cooling, and the heating power of the reactor jacket is reduced at the same time. When the temperature is 0.3℃ lower than the set value, the flow of cold water in the inner coil is turned off to reduce the cooling rate and increase the heating power of the reactor jacket to ensure precise temperature control. During the stable period, the microorganisms mainly synthesize polysaccharides, and their metabolic heat production is stable. The temperature is then switched to the jacketed temperature control of the reactor body, with the inner coil as a backup, to maintain temperature fluctuations ≤ ±0.3℃. In step six, the stirring speed is adjusted as follows: During the logarithmic phase, microorganisms require high oxygen transfer. Based on the dissolved oxygen level detected by the online detection module, when the dissolved oxygen level is below 20% saturation, the stirring speed is automatically increased, and sterile air is injected simultaneously. During the stationary phase, polysaccharides begin to accumulate. The viscosity sensor of the online detection module monitors the viscosity of the fermentation broth in real time. When the viscosity is ≥500 cP, the stirring speed is automatically reduced to reduce shear damage to polysaccharide molecules and ensure the stability of polysaccharide molecular weight.

[0014] Furthermore, the various data detected in step eight are displayed as parameter curves through the visualization interface of the display panel. When the parameters exceed the set range, an audible and visual alarm is automatically issued, and an emergency adjustment program is initiated, such as emergency oxygen supplementation and adjustment of temperature control power. For the control of miscellaneous bacteria, the sterile air filtration effect is continuously monitored during the fermentation process. Every 24 hours, the number of microorganisms at the air filter outlet is tested by sterile sampling. If the number exceeds the standard, the backup filter is automatically switched. The pressure test of the reactor seals is performed weekly to prevent seal failure and subsequent intrusion of miscellaneous bacteria. In step nine, the polysaccharide fermentation broth is obtained. The fermentation endpoint is determined by comprehensively considering the following parameters: polysaccharide concentration, which is monitored using an existing online refractometer. When the polysaccharide concentration increases by ≤0.5g / L for 2 consecutive hours, the turbidity of the cell count remains stable at 1200-1500 NTU for 2 consecutive hours, and the pH of the fermentation broth naturally decreases to 5.5-6.0 or 6.0-6.5 and no longer changes. After all parameters reach the endpoint, the reactor and a series of controls are shut down, and the fermentation broth is discharged and transported to the subsequent extraction process. At the same time, the reactor is preliminarily cleaned to prepare for the next batch of fermentation.

[0015] This invention provides an intelligent temperature-controlled stirred reactor and fermentation method for microbial polysaccharide fermentation, which has the following beneficial effects: In use, this invention raises the base plate, causing the rotating tube and its outer components to move upwards. The guide block and guide ring cause the opening of the sealed pipe to coincide with the opening of the delivery pipe. With the rotating tube slowly rotating inside the reactor, the nutrient solution is evenly delivered to different positions inside the reactor through the delivery pipe and the opening on the sealed pipe. Delivery at different positions effectively enhances the mixing effect between the fermentation broth and the supplementary nutrient solution. At the same time, the rotating tube rotates slowly and continuously during the delivery of the nutrient solution, avoiding the strong shear force that may be generated by rapid stirring, reducing the risk of shear damage to polysaccharide molecules, which is conducive to maintaining the stability of polysaccharide molecular weight and ensuring the quality of the microbial polysaccharides produced by fermentation.

[0016] In addition, by combining multi-layer temperature control and control algorithms with the inner coil and the reaction vessel jacket, precise temperature control in stages is achieved with fluctuations of ≤±0.5℃. This solves the problem of temperature imbalance caused by microbial metabolic heat production, adapts to the needs of different fermentation stages, ensures oxygen transfer efficiency in the logarithmic phase, and avoids polysaccharide shear damage in the stationary phase, thus creatively improving polysaccharide yield and quality.

[0017] In addition, the segmented sterilization process was applied to the fermentation of microbial polysaccharides, which takes into account both the sterilization effect and the retention of nutrients in the culture medium.

[0018] This process increases polysaccharide yield by 15-25% compared to traditional methods. For example, xanthan gum yield increases from 20g / L to 25-30g / L. The polysaccharide purity is ≥92%, and the contamination rate of miscellaneous bacteria is ≤0.5%. The process is highly automated, reducing manual intervention by more than 60% and energy consumption by 10-15%. It is compatible with the fermentation of polysaccharides from various microorganisms and can be applied to the production of xanthan gum, gellan gum, hyaluronic acid, etc. by adjusting parameters, making it highly versatile. Attached Figure Description

[0019] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings of the embodiments will be briefly described below.

[0020] The accompanying drawings described below are only related to some embodiments of the invention and are not intended to limit the invention.

[0021] In the attached diagram: Figure 1 A schematic diagram of the overall structure of the present invention is shown; Figure 2 A schematic cross-sectional view of the reaction vessel structure of the present invention is shown; Figure 3 A schematic cross-sectional view of the guide ring structure of the present invention is shown; Figure 4 A schematic cross-sectional view of the sealing pipe structure of the present invention is shown; Figure 5 The invention is shown by Figure 4 A schematic diagram of the enlarged structure of part A is shown. Figure 6 A three-dimensional structural diagram of the inner coil of the present invention is shown; Figure 7 A three-dimensional structural diagram of the water pipe of the present invention is shown; Figure 8 A cross-sectional view of the positioning cylinder of the present invention is shown; Figure 9 A flowchart of the fermentation method of the present invention is shown.

[0022] List of reference numerals 1. Reactor body; 101. Guide ring; 102. Positioning rod; 103. Traction base plate; 104. Rotating tube; 105. Feeding pipe; 106. Sealing pipe; 107. Guide block; 108. Feed hood; 109. Flow guide hood; 1010. Nutrient solution storage tank; 1011. Flow guide pipe; 2. Inner coil; 201. Diverter pipe; 202. Injection pipe; 203. Multifunctional injection pipe; 204. Mixing blade; 205. Air inlet pipe; 206. Conveying pipe; 207. Aeration pipe; 3. Positioning cylinder; 301. Sealing plate; 302. Push plate; 303. Movable tube; 304. Nozzle; 305. Purification drive pump; 306. Water flow pipe. Detailed Implementation

[0023] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present 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 the present invention. Based on the described 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.

[0024] Please refer to Figures 1 to 9 : Example 1: This invention proposes an intelligent temperature-controlled stirred reactor and fermentation method for microbial polysaccharide fermentation, comprising: a reactor body 1, with two guide rings 101 installed on the inner side of the reactor body 1, wherein the bottom of the guide rings 101 has an inclined structure; two positioning vertical rods 102 are installed on the bottom of the reactor body 1, wherein the bottom of the positioning vertical rods 102 is provided with limiting protrusions; a drive cylinder is installed on the outer bottom of the reactor body 1, and a traction base plate 103 is installed on the output end of the bottom of the drive cylinder; the traction base plate 103 is slidably installed on the outer side of the two positioning vertical rods 102, and the bottom of the traction base plate 103 is... Equipped with a servo motor, a gear is mounted on the output end of the servo motor; a rotating tube 104 is slidably mounted in the middle of the reactor body 1; a gear is mounted at the bottom of the rotating tube 104, penetrating the bottom of the reactor body 1, and meshes with the gear at the output end of the servo motor at the bottom of the traction base plate 103; the bottom of the rotating tube 104 is rotatably mounted on the traction base plate 103; multiple evenly distributed delivery pipes 105 are installed on the outer side of the rotating tube 104; multiple evenly distributed openings are provided on the outer side of the delivery pipes 105; a sealing pipe 106 is slidably mounted on the outer side of the delivery pipes 105, and the side end of the delivery pipes 105... A spring is installed, and the side end of the delivery pipe 105 is connected to the inner end of the sealing pipe 106 via the spring; the outer side of the sealing pipe 106 has multiple evenly distributed openings, which can coincide with the openings on the outer side of the delivery pipe 105, and a guide block 107 is installed at the outer end of the sealing pipe 106; the guide block 107 is slidably installed on the inner side of the guide ring 101; the reactor body 1 has a cylindrical structure, with a feed inlet at the top, a display panel installed on the outer side of the reactor body 1, and a sandwich structure on the side wall of the reactor body 1. A connecting flange is installed on the outer side of the reactor body 1, and the bottom of the reactor body 1... Support legs are installed at the edge of the reactor vessel 1; an online detection module is installed on the outside of the rotating tube 104, which integrates a pH sensor, dissolved oxygen electrode, temperature sensor, viscosity sensor and turbidity sensor; a feed hood 108 is installed at the top of the rotating tube 104 through the top of the reactor vessel 1; a flow guide hood 109 is rotatably installed on the outside of the feed hood 108; a nutrient solution tank 1010 is installed on the outside of the reactor vessel 1; a drive pump is installed on the top of the nutrient solution tank 1010, and a flow guide pipe 1011 is installed on the output end of the drive pump; the side end of the flow guide pipe 1011 is connected to the outside of the flow guide hood 109.

[0025] In this embodiment of the invention, during the fermentation of microbial polysaccharides, the fermentation culture medium and microbial seed liquid are placed inside the reactor body 1 through the feed inlet at the top of the reactor body 1. The gear at the output end of the servo motor on the traction base plate 103 drives the gear at the bottom of the rotating tube 104 to rotate. In turn, the rotating tube 104 drives the outer feeding pipe 105, the outer sealing pipe 106, and the stirring plate 204 to rotate for stirring. When nutrient solution needs to be added after fermentation, the drive cylinder at the bottom of the outer side of the reactor body 1 drives the traction base plate 103 to move upward along the two positioning vertical rods 102. The traction base plate 103 drives the rotating tube 104 to move upward inside the reactor body 1. The outer sealing pipe 106 of the outer feeding pipe 105 of the rotating tube 104 moves upward. The guide block 107 at the outer end of the sealing pipe 106 abuts against the inner side of the guide ring 101, so that the guide block 107 drives the sealing pipe 106 slides on the delivery pipe 105 so that the opening on the sealed pipe 106 coincides with the opening on the delivery pipe 105. The drive pump on the nutrient solution storage tank 1010 draws the internally stored nutrient solution and flows it through the guide pipe 1011 into the guide hood 109. The nutrient solution then enters the rotating tube 104 through the feed hood 108. The nutrient solution then flows out through the openings on the delivery pipe 105 and the sealed pipe 106. The rotating tube 104 continues to rotate slowly inside the reactor body 1. The feed hood 108 at the top of the rotating tube 104 rotates inside the guide hood 109, so that the nutrient solution is evenly delivered into the reactor body 1 at different positions and fully mixed with the fermentation liquid. This improves the mixing effect of the fermentation liquid and the supplemented nutrient solution. Furthermore, the slow rotation of the rotating tube 104 will not cause shear damage to the polysaccharide molecules that have already been produced. It continuously provides nutrient solution for the fermentation of the fermentation liquid and ensures the stability of the polysaccharide molecular weight.

[0026] In Example 2, based on Example 1, multiple inner coils 2 are installed inside the reactor body 1; a diversion pipe 201 is installed on the outer side of each inner coil 2; an injection pipe 202 is installed through the side end of each diversion pipe 201, penetrating the side of the reactor body 1; a connecting flange is installed on the top of the injection pipe 202; multiple multi-functional feeding pipes 203 are installed on the top of the reactor body 1; a stirring blade 204 is installed on the outer side of the rotating pipe 104; an air inlet pipe 205 is installed in the jacket of the reactor body 1; a quick connector is installed at the outer end of the air inlet pipe 205, and multiple conveying pipes 206 are installed on the side of the air inlet pipe 205; the conveying pipes 206 have an annular structure, and the side of the conveying pipes 206... An aeration pipe 207 is provided on the surface; the aeration pipe 207 is located inside the reactor body 1; a positioning cylinder 3 is installed on one side of the top of the reactor body 1; a normally closed sealing plate 301 is rotatably installed at the bottom of the positioning cylinder 3, and a spring is installed at the top of the positioning cylinder 3, and a push plate 302 is installed at the top of the spring; a movable pipe 303 is installed on the side end of the push plate 302; a nozzle 304 in different directions is installed through the bottom of the movable pipe 303 through the top of the positioning cylinder 3; the bottom of the movable pipe 303 and the nozzle 304 are located inside the positioning cylinder 3; a purification type drive pump 305 is installed on one side of the top of the reactor body 1; a water flow pipe 306 is installed on the output end of the top of the purification type drive pump 305;The side end of the water pipe 306 is connected to the top of the movable pipe 303. During the fermentation of microbial polysaccharides, before adding the culture medium, the push plate 302 is pressed downwards from the top of the positioning cylinder 3. The spring at the bottom of the push plate 302 contracts, and the side end of the push plate 302 drives the movable pipe 303 to move downwards inside the positioning cylinder 3. The bottom of the movable pipe 303 pushes the two normally closed sealing plates 301 outwards, so that the nozzle 304 on the outer side of the bottom of the movable pipe 303 is inside the reactor body 1. Then, the purification-type drive pump 305 can... Water is drawn from the outside environment and treated by the purification pump 305 before flowing through the water pipe 306 into the interior of the movable pipe 303. The water is then sprayed through nozzles 304, and the high-pressure water jets from the nozzles 304 in different directions can clean different locations inside the reactor body 1, ensuring the cleanliness of the reactor body 1 and other components. During the high-temperature sterilization stage, high-temperature steam is diverted through the injection pipe 202 into the diversion pipe 201 and then into the interior of the inner coil 2. At the same time, the high-temperature steam is held in place by the reactor body 1. The connecting flange enters the jacketed interior to sterilize the interior of the reactor body 1 at high temperature, preventing contaminants from affecting the quality of microbial polysaccharide fermentation. The online detection module on the rotating tube 104 allows for temperature adjustment of the inner coil 2 and the jacket of the reactor body 1 at different fermentation stages, ensuring fermentation quality. For pH adjustment, NaOH or H2SO4 solution can be added through the multi-functional injection pipe 203 on the reactor body 1. For dissolved oxygen adjustment, the outer end of the air inlet pipe 205 is connected to an external oxygen source, allowing sterile oxygen to be distributed to the interior of each conveying pipe 206. Multiple aeration pipes 207 are installed on the conveying pipes 206 at different heights, allowing gas to enter the liquid in a dispersed manner. Each aeration pipe 207 supplies gas at a relatively low gas linear velocity, avoiding the formation of high-speed gas jets. By extending the effective residence path of bubbles in the liquid, the utilization efficiency of oxygen in high-viscosity liquids is improved, achieving the oxygen supply required for polysaccharide fermentation without significantly increasing the stirring speed. A method for fermenting microbial polysaccharides includes the following steps: First, clean the reactor body. Use high-pressure water to rinse the inner wall of the reactor body 1, the stirring plate 204, the feeding pipe 105 and the sealing pipe 106 to remove residual impurities. Then, immerse it in sodium hydroxide solution for 30-40 minutes, and then rinse it with deionized water until the pH is neutral. Second, staged sterilization: First, sterilize the inside of the reactor body 1 with high temperature of saturated steam, then inject fermentation medium into the reactor body 1, obtain clean medium after high temperature sterilization, and then cool it down to the fermentation start temperature. 3. Activation and inoculation of microbial strains: Inject the pre-cultured microbial seed liquid into reaction vessel 1; Fourth, for the initial parameter settings, the temperature, stirring speed, pH value, and dissolved oxygen value are adjusted according to the type of microorganism. Fifth, phased intelligent temperature control: the fermentation stages are divided into the lag phase, logarithmic phase and stationary phase, and temperature is controlled at different stages. Sixth, by adjusting the rotation speed of the stirring plate 204 at different fermentation stages, and in conjunction with the amount of oxygen injected, the dissolved oxygen content and viscosity of the fermentation broth are adjusted. 7. Nutrient supplementation and regulation: The amount of cells is monitored by the turbidity sensor on the online detection module. When the turbidity reaches 800-1000 NTU, the drive pump on the nutrient solution storage tank 1010 is turned on to add carbon and nitrogen sources to the reactor body 1 to avoid substrate depletion and fermentation stagnation. 8. During the fermentation process monitoring and miscellaneous microbial control stage, the online detection module collects real-time data on pH, dissolved oxygen, temperature, viscosity, and turbidity. The number of microorganisms is detected by injecting air into a multi-functional feeding pipe 203. At the same time, the sealing condition of the reactor body 1 is checked regularly to reduce the intrusion of miscellaneous microorganisms. 9. By detecting that the polysaccharide concentration, cell count, and pH value of the fermentation broth are in a stable state, a stable polysaccharide fermentation broth is obtained. Step two, segmented sterilization, includes: In the first stage, 121°C saturated steam is introduced into the jacket and inner coil of the reactor, maintaining a pressure of 0.12-0.15 MPa for 30-40 minutes to sterilize and kill microorganisms on the surface of the reactor. In the second stage, the prepared fermentation medium is injected into the reactor body 1, and 115℃ saturated steam is introduced through the inner coil 2 to maintain the pressure at 0.08-0.1MPa for 20-25 minutes to sterilize and avoid high temperature from destroying the nutrients in the medium. In the third stage, after sterilization, cooling is carried out. Cooling water is introduced through the jacket of the inner wall of the reactor body 1 to reduce the temperature of the culture medium to the fermentation start temperature. At the same time, sterile air is introduced into the reactor body 1 to prevent the invasion of other bacteria. In step three, the preserved microbial strains are inoculated into the seed culture medium and cultured in a shaker for 12-16 hours in advance to the logarithmic phase to obtain the seed liquid. The seed liquid is then injected into reaction vessel 1 to avoid contamination during the inoculation process. In step four, the temperature is set according to the type of microorganism, for example, xanthan gum fermentation at 28-30℃ and gellan gum fermentation at 30-32℃. The stirring speed of the stirring plate 204 is set to 150-200 rpm to ensure uniform dispersion of the bacteria. The pH value is set according to the type of microorganism, for example, 6.5-7.0 for xanthan gum fermentation and 7.0-7.5 for gellan gum fermentation. It is adjusted by using an alkali or acid addition device through a multi-functional feeding pipe 203. The dissolved oxygen content is maintained at 20-30% saturation by adjusting the stirring speed and the flow rate of sterile air. Step five, segmented temperature control, includes: During the delayed period, microbial metabolic heat production is low. Temperature control is mainly achieved through the jacket of the reactor body 1, with the inner coil 2 providing auxiliary control, to maintain temperature fluctuations ≤ ±0.5℃. During the logarithmic phase, microorganisms multiply rapidly, and metabolic heat production increases sharply. The temperature inside reactor 1 is collected by the online detection module. When the temperature is 0.3℃ higher than the set value, cold water is injected into the inner coil 2 for cooling, and the heating power of the jacket of reactor 1 is reduced. When the temperature is 0.3℃ lower than the set value, the flow of cold water in the inner coil 2 is turned off to reduce the cooling rate and increase the heating power of the jacket of reactor 1 to ensure precise temperature control. During the stable period, the microorganisms mainly synthesize polysaccharides, and the metabolic heat production is stable. The temperature is then switched to the jacket of the reactor body 1 for control, with the inner coil 2 as a backup, to maintain temperature fluctuations ≤ ±0.3℃. In step six, the stirring speed of the stirring plate 204 is adjusted. During the logarithmic phase, microorganisms require high oxygen transfer. Oxygen is injected into the liquid inside the reactor body 1 through the aeration pipe 207. Based on the dissolved oxygen level detected by the online detection module, when the dissolved oxygen level is lower than 20% saturation, the stirring speed is automatically increased in conjunction with the oxygen injected through the aeration pipe 207 to increase the dissolved oxygen level. At the same time, sterile air flow is injected. During the stationary phase, polysaccharides begin to accumulate. The viscosity sensor of the online detection module monitors the viscosity of the fermentation broth in real time. When the viscosity is ≥500 cP, the stirring speed is automatically reduced to reduce shear damage to polysaccharide molecules and ensure the stability of polysaccharide molecular weight. The various data detected in step eight are displayed as parameter curves through the visual interface of the display panel. When the parameters exceed the set range, an audible and visual alarm is automatically issued, and an emergency adjustment program is initiated, such as emergency oxygen supplementation and adjustment of temperature control power. For the control of miscellaneous bacteria, the sterile air filtration effect is continuously monitored during the fermentation process. Every 24 hours, the number of microorganisms at the air filter outlet is tested by sterile sampling. If the number exceeds the standard, the backup filter is automatically switched. The pressure test of the reactor seals is performed weekly to prevent seal failure and subsequent intrusion of miscellaneous bacteria. In step nine, the polysaccharide fermentation broth is obtained. The fermentation endpoint is determined by comprehensively considering the following parameters: polysaccharide concentration, which is monitored using an online refractometer. When the polysaccharide concentration increases by ≤0.5g / L for 2 consecutive hours, the turbidity of the cell mass remains stable at 1200-1500 NTU for 2 consecutive hours, and the pH value of the fermentation broth naturally decreases to 5.5-6.0 or 6.0-6.5 and no longer changes. After all parameters reach the endpoint, reactor 1 and a series of controls are shut down, and the fermentation broth is discharged and transported to the subsequent extraction process. At the same time, reactor 1 is preliminarily cleaned to prepare for the next batch of fermentation.

[0027] In Example 3, based on Example 1, for xanthan gum fermentation, firstly, high-pressure water jets are sprayed from nozzle 304 at the bottom of the movable tube 303 to rinse the inside of the reactor body 1 and various components. Then, a 1.5% NaOH solution at 55°C is added and soaked for 35 minutes. Next, it is rinsed with deionized water until a neutral pH is reached. High-temperature steam at 121°C and 0.13 MPa is injected into the jacket between the inner coil 2 and the reactor body 1 for sterilization for 35 minutes. After injecting the culture medium, it is sterilized again with steam at 115°C and 0.09 MPa for 22 minutes. The mixture is then cooled to 29°C, sterile air is introduced, and a positive pressure of 0.025 MPa is maintained. Pre-cultured Xanthomonas oryzae seed solution is injected into the reactor body 1 at a 6% inoculation rate. The initial parameters are set as follows: temperature 29°C, stirring speed of stirrer 204 180 rpm, pH of fermentation broth 6.8. The dissolved oxygen content was 25%. During fermentation, temperature was controlled during the delayed phase, with temperature fluctuations of ±0.4℃. The stirring plate 204 rotated at 180 rpm. During the logarithmic phase, real-time temperature control was implemented. When the temperature exceeded 29.3℃, the inner coil 2 was turned on for cooling. When the dissolved oxygen content was below 20%, the stirring plate 204 was rotated at 250 rpm and the air flow rate was 1.2 vvm. When the turbidity reached 900 NTU, glucose and peptone were extracted through the nutrient solution tank 1010 and injected into the interior of the reactor body 1. During the steady-state phase, the temperature was controlled in the jacket of the reactor body 1. When the viscosity reached 500 cP, the stirring plate 204 was rotated at 120 rpm. Fermentation was stopped when the polysaccharide concentration was 28.5 g / L, the turbidity was 1350 NTU, and the pH was 5.8 after 72 hours. A fermentation broth with a polysaccharide purity of 93.2% and a contamination rate of 0.3% was obtained. High-quality xanthan gum was then extracted from the broth.

[0028] The working principle of this embodiment is as follows: Pressing the push plate 302 downwards on the top of the positioning cylinder 3 drives the movable tube 303 to move downwards within the positioning cylinder 3. The bottom of the movable tube 303 pushes the two normally closed sealing plates 301 outwards. The nozzle 304 on the outer side of the bottom of the movable tube 303 is located inside the reactor body 1. The purification-type drive pump 305 draws external water, which, after treatment, flows through the water pipe 306 into the movable tube 303 and is then sprayed through the nozzle 304. The high-pressure water jets sprayed from the nozzles 304 in different directions can clean the inner wall of the reactor body 1 and other components. High-temperature steam is diverted through the injection pipe 202 into the diversion pipe 201 and then into the inner coil 2. At the same time, high-temperature steam enters the jacket through the connecting flange clamped by the reactor body 1 to sterilize the interior of the reactor body 1 at high temperature. The online detection module on the rotating tube 104 allows the inner coil 2 and the jacket of the reactor body 1 to control the temperature at different fermentation stages. The culture medium and microbial seed liquid used for fermentation are placed inside the reactor body 1 through the feed inlet at the top of the reactor body 1. The servo motor on the traction base plate 103 drives the rotating tube 104 to rotate the outer feeding pipe 105 and the stirring plate 204 for stirring. When nutrient solution needs to be added after fermentation, the drive cylinder at the bottom of the outer side of the reactor body 1 drives the traction base plate 103 to move upward along the two positioning vertical rods 102. The traction base plate 103 drives the sealing pipe 106 outside the feeding pipe 105 outside the rotating tube 104 to move upward. The guide block 107 at the outer end of the sealing pipe 106 abuts against the inner side of the guide ring 101. The sealing pipe 106 is slid on the delivery pipe 105 by the block 107, so that the openings of the two coincide. The drive pump on the nutrient solution storage tank 1010 draws the nutrient solution through the guide pipe 1011 into the guide shroud 109, and then through the feed shroud 108 into the rotating pipe 104. The nutrient solution then flows out through the openings on the delivery pipe 105 and the sealing pipe 106. The nutrient solution is evenly delivered into the interior of the reactor body 1 at different locations and fully mixed with the fermentation liquid, thereby improving the mixing effect of the fermentation liquid and the supplemented nutrient solution.

[0029] The following points should be noted in this article: 1. The accompanying drawings of the embodiments of the present invention only involve the structures involved in the embodiments of the present invention; other structures can refer to general designs.

[0030] 2. Where there is no conflict, the embodiments of the present invention and the features thereof can be combined with each other to obtain new embodiments.

[0031] The above are merely specific embodiments of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A smart temperature-controlled stirred reactor for microbial polysaccharide fermentation, comprising: The reactor body (1) has two guide rings (101) installed on its inner side; characterized in that two positioning rods (102) are installed at the bottom of the reactor body (1); a drive cylinder is installed at the bottom of the outer side of the reactor body (1), and a traction base plate (103) is installed on the output end of the bottom of the drive cylinder; the traction base plate (103) is slidably installed on the outer side of the two positioning rods (102), and a servo motor is installed at the bottom of the traction base plate (103), and a gear is installed on the output end of the servo motor; a rotating tube (104) is slidably installed at the middle position of the reactor body (1); a gear is installed at the bottom of the rotating tube (104) penetrating the bottom of the reactor body (1), and the gear meshes with the gear at the output end of the servo motor at the bottom of the traction base plate (103). The bottom of the rotating tube (104) is rotatably mounted on the traction base plate (103); multiple evenly distributed delivery pipes (105) are installed on the outer side of the rotating tube (104); multiple evenly distributed openings are provided on the outer side of the delivery pipes (105); a sealing pipe (106) is slidably installed on the outer side of the delivery pipes (105), and a spring is installed on the side end of the delivery pipes (105); the side end of the delivery pipes (105) is connected to the inner end of the sealing pipes (106) through the spring; multiple evenly distributed openings are provided on the outer side of the sealing pipes (106), and the openings can coincide with the openings on the outer side of the delivery pipes (105); a guide block (107) is installed on the outer end of the sealing pipes (106); the guide block (107) is slidably mounted on the inner side of the guide ring (101).

2. The intelligent temperature-controlled stirred reactor for microbial polysaccharide fermentation according to claim 1, characterized in that, The reactor body (1) has a feed inlet at the top, a display panel on the outside of the reactor body (1), and a sandwich structure on the side wall of the reactor body (1). A connecting flange is installed on the outside of the reactor body (1), and a support leg is installed at the bottom edge of the reactor body (1). An online detection module is installed on the outside of the rotating tube (104), which integrates a pH sensor, dissolved oxygen electrode, temperature sensor, viscosity sensor, and turbidity sensor. A feed hood (108) is installed at the top of the rotating tube (104) through the top of the reactor body (1).

3. The intelligent temperature-controlled stirred reactor for microbial polysaccharide fermentation according to claim 2, characterized in that, A flow guide (109) is rotatably installed on the outside of the feed hood (108); a nutrient solution tank (1010) is installed on the outside of the reactor body (1); a drive pump is installed on the top of the nutrient solution tank (1010), and a flow guide pipe (1011) is installed on the output end of the drive pump; the side end of the flow guide pipe (1011) is connected to the outside of the flow guide (109).

4. The intelligent temperature-controlled stirred reactor for microbial polysaccharide fermentation according to claim 3, characterized in that, The reactor body (1) has multiple inner coils (2) installed on its inner side; each inner coil (2) has a diversion pipe (201) installed on its outer side; the side ends of the diversion pipes (201) are connected to an injection pipe (202) through the side of the reactor body (1); the top of the injection pipe (202) is fitted with a connecting flange; the top of the reactor body (1) has multiple multi-functional injection pipes (203); the outside of the rotating pipe (104) is fitted with a stirring blade (204); an air inlet pipe (205) is installed in the jacket of the reactor body (1); the outer end of the air inlet pipe (205) is fitted with a quick connector, and the side of the air inlet pipe (205) has multiple conveying pipes (206); the side of the conveying pipe (206) is fitted with an aeration pipe (207); the aeration pipe (207) is located inside the reactor body (1).

5. The intelligent temperature-controlled stirred reactor for microbial polysaccharide fermentation according to claim 4, characterized in that, A positioning cylinder (3) is installed on one side of the top of the reactor body (1); a normally closed sealing plate (301) is rotatably installed at the bottom of the positioning cylinder (3), and a spring is installed at the top of the positioning cylinder (3), and a push plate (302) is installed at the top of the spring; a movable tube (303) is installed at the side end of the push plate (302).

6. The intelligent temperature-controlled stirred reactor for microbial polysaccharide fermentation according to claim 5, characterized in that, The bottom of the movable tube (303) passes through the top of the positioning cylinder (3) and is equipped with nozzles (304) in different directions; the bottom of the movable tube (303) and the nozzles (304) are located inside the positioning cylinder (3); a purification type drive pump (305) is installed on one side of the top of the reactor body (1); a water flow pipe (306) is installed on the output end of the top of the purification type drive pump (305); the side end of the water flow pipe (306) is connected to the top of the movable tube (303).

7. A method for microbial polysaccharide fermentation as described in any one of claims 1 to 6, characterized in that, Includes the following steps: First, the inner wall of the reactor body (1), the stirring plate (204), the feeding pipe (105) and the sealing pipe (106) are rinsed with high pressure water to remove residual impurities. Then, sodium hydroxide solution is passed through for soaking, and then deionized water is used to rinse until the pH is neutral. Second, the process involves staged sterilization. First, the inside of the reactor body (1) is sterilized using the high temperature of saturated steam. Then, fermentation medium is injected into the reactor body (1). After sterilization with high temperature, a clean medium is obtained and then cooled to the fermentation start temperature. Third, inject the pre-cultured microbial seed liquid into the reaction vessel body (1); Fourth, adjust the parameters of temperature, stirring speed, pH value, and dissolved oxygen according to the type of microorganism; Fifth, the fermentation process is divided into three stages: the delayed phase, the logarithmic phase, and the stationary phase, with temperature control implemented at each stage. Sixth, by adjusting the rotation speed of the stirring plate (204) at different fermentation stages, combined with the amount of oxygen injected, the dissolved oxygen content and viscosity of the fermentation broth are adjusted.

7. Monitor the amount of bacteria by using the turbidity sensor on the online detection module, turn on the drive pump on the nutrient solution tank (1010), and add carbon and nitrogen sources to the reactor body (1); 8. The online detection module collects pH, dissolved oxygen, temperature, viscosity and turbidity data in real time, and at the same time checks the sealing of the reaction vessel (1) regularly to reduce the intrusion of miscellaneous bacteria; 9. By detecting that the polysaccharide concentration, cell volume, and pH value of the fermentation broth are in a stable state, a stable polysaccharide fermentation broth is obtained.

8. The microbial polysaccharide fermentation method according to claim 7, characterized in that, The segmented sterilization in step two includes: In the first stage, 121°C saturated steam is introduced into the jacket and inner coil of the reactor for sterilization for 30-40 minutes to kill microorganisms on the surface of the reactor. In the second stage, the prepared fermentation culture medium is injected into the reactor body (1), and 115℃ saturated steam is introduced through the inner coil (2) for sterilization for 20-25 minutes. In the third stage, after sterilization, cooling is carried out. Cooling water is introduced through the interlayer of the inner wall of the reactor body (1) to reduce the temperature of the culture medium to the fermentation start temperature. At the same time, sterile air is introduced into the reactor body (1). In step three, the preserved microbial strains are inoculated into the seed culture medium and cultured in a shaker for 12-16 hours in advance to the logarithmic phase to obtain the seed liquid. The seed liquid is then injected into the reaction vessel (1). In step four, the parameters are set such that the pH value is set according to the type of microorganism and is adjusted by using an alkali or acid addition device through a multi-functional injection pipe (203). The dissolved oxygen content is maintained at 20-30% saturation and controlled by adjusting the stirring speed and the flow rate of sterile air.

9. The microbial polysaccharide fermentation method according to claim 8, characterized in that, The segmented temperature control in step five includes: During the delayed period, microbial metabolism generates less heat, so the temperature is mainly controlled by the jacket of the reactor body (1), with the inner coil (2) as an auxiliary measure, to maintain temperature fluctuations ≤ ±0.5℃; During the logarithmic phase, microorganisms multiply rapidly, and metabolic heat production increases sharply. The temperature inside the reactor body (1) is collected by the online detection module. When the temperature is higher than the set value by 0.3℃, cold water is injected into the inner coil (2) for cooling, and the heating power of the reactor body (1) jacket is reduced. When the temperature is lower than the set value by 0.3℃, the flow of cold water in the inner coil (2) is turned off to reduce the cooling rate and increase the heating power of the reactor body (1) jacket to ensure accurate temperature control. During the stable period, the microorganisms mainly synthesize polysaccharides and have stable metabolic heat production. The temperature is then switched to the jacket of the reactor body (1) for temperature control, with the inner coil (2) as a backup, to maintain temperature fluctuations ≤ ±0.3℃. In step six, the stirring speed of the stirring plate (204) is adjusted. During the logarithmic phase, microorganisms require high oxygen transfer. Based on the dissolved oxygen level detected by the online detection module, when the dissolved oxygen level is lower than 20% saturation, the stirring speed is automatically increased, and sterile air is injected. During the stationary phase, polysaccharides begin to accumulate. The viscosity sensor of the online detection module monitors the viscosity of the fermentation broth in real time. When the viscosity is ≥500 cP, the stirring speed is automatically reduced.

10. A method for fermenting microbial polysaccharides according to claim 9, characterized in that, The various data detected in step eight are displayed as parameter curves through the visualization interface of the display panel. When the parameters exceed the set range, an audible and visual alarm is automatically issued, and an emergency adjustment program is initiated. For the control of miscellaneous bacteria, the sterile air filtration effect is continuously monitored during the fermentation process. Every 24 hours, the number of microorganisms at the air filter outlet is detected by sterile sampling. If the number exceeds the standard, the backup filter is automatically switched. The pressure test of the reactor seal is performed weekly.

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