Preparation process of high-efficiency microbial degradation liquefied cellulose complex microbial agent

Abstract

This invention discloses a highly efficient microbial process for preparing liquefied cellulose composite inoculants, relating to the field of microbial acoustic magnetic field treatment. The process includes the following steps: Step 1, inoculum activation and seed culture preparation; Step 2, inoculation and fermentation; Step 3, multi-field coupling enhancement and online detection; Step 4, intelligent feedback control; and Step 5, inoculum post-treatment. The intelligent multi-field coupled reactor includes a tank for carrying the inoculum, an ultrasonic stirring device located within the tank, a magnetic field generating device located on the outer wall of the tank, and online viscometers, fluorescent dissolved oxygen sensors, and near-infrared enzyme activity sensors for detecting the inoculum within the tank. The preparation process provided by this invention, by constructing an intelligent fermentation system with multi-field coupling of ultrasound and magnetic fields, fundamentally solves the technical defects of traditional cellulose-degrading inoculants, such as long fermentation cycles and low enzyme activity.

Description

Technical Field

[0001] This invention mainly relates to the technical field of microbial acoustic magnetic field treatment, specifically a process for preparing a highly efficient microbial degradation liquefied cellulose composite agent. Background Technology

[0002] With the increasing severity of the global energy crisis and environmental pollution, the conversion of renewable biomass resources into clean energy has become a research hotspot. Cellulose, as the most abundant renewable resource in nature, is a key link in the efficient degradation into fermentable sugars for the production of biofuels and chemical products. Biodegradation methods are favored due to their mild conditions and environmental friendliness. The performance of cellulose-degrading microbial agents directly determines the bioconversion efficiency and economic feasibility. Developing efficient and stable compound microbial agent preparation technologies is of great significance for promoting the industrialization of cellulosic ethanol.

[0003] Currently, the preparation of cellulose-degrading microbial agents mainly employs liquid submerged fermentation. Traditional methods typically control only conventional parameters such as temperature, stirring speed, and aeration rate, using a single microorganism or a simple mixture of strains for fermentation. However, cellulose-degrading bacteria are mostly filamentous fungi, and the fermentation process involves complex metabolic regulation of mycelial growth and product synthesis, long fermentation cycles, and low enzyme activity. Although existing technologies attempt to improve enzyme production capacity through culture medium optimization or mutagenesis breeding, it remains difficult to overcome the metabolic bottleneck between cell growth and product synthesis. Furthermore, the lack of precise control methods in the fermentation process results in poor viable cell count and enzyme activity stability, making it difficult to meet industrial-scale cellulose degradation efficiency requirements.

[0004] To address the aforementioned technical deficiencies, this invention aims to provide a highly efficient process for preparing a microbial degradation liquefied cellulose composite inoculant, thereby solving the technical problems of long fermentation cycles, low cellulase activity, and poor inoculant stability in traditional fermentation processes, and achieving efficient and stable preparation of cellulose degradation inoculants. Summary of the Invention

[0005] Based on this, the purpose of the present invention is to provide a highly efficient microbial degradation liquefied cellulose composite inoculant preparation process to solve the technical problems mentioned in the background art.

[0006] To achieve the above objectives, the present invention provides the following technical solution: a high-efficiency microbial degradation liquefied cellulose composite inoculant preparation process, comprising the following steps: Step 1, inoculum activation and seed liquid preparation: after activating the cellulose-degrading inoculum, it is inoculated into a seed culture medium for cultivation to obtain seed liquid one; Step 2, inoculation and fermentation: seed liquid one is inoculated into a fermentation medium and fermented in an intelligent multi-field coupled reactor; Step 3, multi-field coupling enhancement and online detection: during fermentation, an ultrasonic stirring device applies an ultrasonic field, and a magnetic field generator applies a magnetic field to achieve synergistic enhancement, and an online viscometer and a fluorescence dissolved oxygen sensor are used to detect the enhanced field. The fermentation parameters are detected in real time by a near-infrared enzyme activity sensor; Step 4: Intelligent feedback control, based on the online detection results of the fermentation parameters, the controller automatically adjusts the ultrasonic intensity of the ultrasonic stirring device, the magnetic field intensity of the magnetic field generator, and the fermentation process parameters; Step 5: Post-treatment of the microbial agent, after fermentation, the fermentation broth is collected and post-treated to obtain the compound microbial agent product; The intelligent multi-field coupled reactor includes a tank for carrying the microbial agent, an ultrasonic stirring device installed in the tank, a magnetic field generator installed on the outer wall of the tank, and an online viscometer, a fluorescence dissolved oxygen sensor, and a near-infrared enzyme activity sensor for detecting the microbial agent in the tank.

[0007] Preferably, in step three, a staged multi-field enhancement strategy is adopted according to the fermentation stage: Lag phase: Magnetic field strength of 50-100 mT is applied, no ultrasound is applied; Early logarithmic phase: Ultrasound strength of 20-30 kHz and magnetic field strength of 100-150 mT are applied; Late logarithmic phase: Ultrasound strength of 30-40 kHz and magnetic field strength of 150-200 mT are applied; Stationary phase: Ultrasound strength of 20-30 kHz and magnetic field strength of 80-120 mT are applied; The ultrasound in the early logarithmic phase is applied intermittently with a duty cycle of 30-50%; the ultrasound in the late logarithmic phase is applied continuously; the ultrasound in the stationary phase is applied intermittently with a duty cycle of 25-35%. In this preferred embodiment, this strategy dynamically matches different field strengths and action modes to the physiological characteristics of different growth stages of microorganisms, achieving precise metabolic flux regulation from "promoting germination" to "enhancing synthesis" and then to "extending the enzyme production cycle," maximizing the yield of cellulase and cell activity.

[0008] Preferably, the ultrasonic stirring device includes a drive turntable rotatably connected to the top of the tank, a rotating tube passing through the drive turntable, multiple stirring boxes hinged at one end to the outer wall of the rotating tube and arranged in a circular array, an ultrasonic generating component disposed within each stirring box, and a power component disposed on the top of the drive turntable for simultaneously driving the multiple stirring boxes to oscillate. In this preferred embodiment, this design deeply integrates traditional mechanical stirring with an ultrasonic transmitting unit, and the stirring boxes can oscillate independently, realizing dynamic directional emission of ultrasonic waves within the reactor, effectively eliminating ultrasonic dead zones, and significantly improving the uniformity of the sound field distribution within the tank.

[0009] Preferably, the ultrasonic generating component includes a cross-shaped plate disposed within the mixing chamber, eight transducers symmetrically disposed at the four ends of the cross-shaped plate, and a wavelength acoustic matching plate with one end abutting against the transducers and the other end abutting against the inner wall of the mixing chamber. In this preferred embodiment, by symmetrically installing eight transducers on the cross-shaped plate and using the wavelength acoustic matching plate for impedance matching, the reflection loss of ultrasonic energy at the interface is minimized, ensuring that the ultrasonic waves are efficiently and uniformly transmitted into the fermentation broth, thereby enhancing the cavitation effect.

[0010] Preferably, the power component includes an annular frame at the top of the mixing box, an electric telescopic cylinder at the top of the drive turntable with its actuating end penetrating the drive turntable, a first lifting ring at the actuating end of the electric telescopic cylinder and sleeved on the outer wall of the rotating tube, multiple vertical plates connected at their top ends to the bottom of the first lifting ring, a second lifting ring connected at its top ends to the bottom ends of the vertical plates and sleeved on the outer wall of the rotating tube, multiple support rods arranged in a ring array on the outer wall of the second lifting ring, and a drive rod with one end connected to the end of the support rod and the other end slidably connected to the inner wall of the annular frame. In this preferred embodiment, the power component, through the electric telescopic cylinder and a multi-stage linkage mechanism, achieves precise synchronous control of the swing angles of multiple mixing boxes.

[0011] Preferably, the device further includes a power supply component located at the top of the tank. The power supply component includes an end cap fixed to the top of the tank via a bracket, a conductive stator located within the end cap, and a rotor conductive ring located on the inner wall of the rotating tube and in contact with the conductive stator. In this preferred embodiment, the rotating conductive ring structure solves the problem of providing stable power to the transducer during dynamic rotation and oscillation, ensuring the reliability of the electrical connection during continuous movement of the equipment and avoiding cable entanglement issues.

[0012] Preferably, the device further includes a cooling device comprising a circulation pump located at the top of the drive turntable, an inlet ring and a outlet ring sequentially arranged from top to bottom on the inner wall of the rotating tube, and a heat-conducting pipe embedded in the cross-shaped plate. The inlet end of the heat-conducting pipe is connected to the inlet ring, and the outlet end of the heat-conducting pipe is connected to the outlet ring. The inlet ring is connected to the output end of the circulation pump via an input pipe, and the outlet ring is connected to the input end of the circulation pump via an output pipe. In this preferred embodiment, the cooling device constructs a closed-loop liquid-cooled circulation from the heat source to the heat exchange components, which can promptly remove the heat generated by the high-power ultrasonic transducer during operation, prevent localized high temperatures from affecting bacterial activity, and ensure temperature stability during long-term fermentation.

[0013] Preferably, the device further includes a heat exchange component, which comprises a heat exchange section disposed on the output pipe, multiple heat exchange fins disposed on the heat exchange section, a mounting cover disposed on the outer wall of the end cover, and multiple thermoelectric cooling chips disposed within the mounting cover. In this preferred embodiment, the introduction of thermoelectric cooling chips in conjunction with the fins for active heat exchange provides higher temperature control accuracy, faster response speed, and a more compact structure compared to traditional cooling medium heat exchange, further enhancing the heat exchange efficiency of the cooling device.

[0014] Preferably, the device further includes a sound-absorbing component disposed on the outer wall of the tank. The sound-absorbing component includes a sound-absorbing cover plate disposed on the outer wall of the tank, and the sound-absorbing cover plate has multiple sound-absorbing grooves on the side near the tank. In this preferred embodiment, by providing a sound-absorbing cover plate with sound-absorbing grooves, the structural noise and reflected waves generated by the ultrasonic field on the tank wall are effectively absorbed and damped, which not only improves the working environment but also reduces noise interference and enhances the purity and stability of the ultrasonic field.

[0015] Preferably, the magnetic field generating device includes an insulating cover plate disposed on the outer wall of the silencing cover plate, multiple through holes passing through the insulating cover plate, a first airflow ring and a second airflow ring respectively disposed on the top and bottom of the insulating cover plate and communicating with the through holes, multiple coil segments disposed on the outer wall of the insulating cover plate, and a shielding cover disposed on the outer wall of the tank and covering the coil segments. In this preferred embodiment, the magnetic field generating device adopts a segmented coil design and integrates an airflow cooling channel, which can not only accurately control the magnetic field intensity in different areas, but also effectively solve the coil heating problem; at the same time, the shielding cover prevents magnetic field leakage, ensuring the strength and directionality of the magnetic field applied to the bacterial agent inside the tank.

[0016] In summary, the present invention has the following main beneficial effects: The efficient microbial degradation liquefied cellulose composite inoculant preparation process provided by this invention fundamentally solves the technical defects of traditional cellulose degradation inoculants, such as long fermentation cycle and low enzyme activity, by constructing an intelligent fermentation system with multi-field coupling of ultrasound and magnetic field. The core of this process lies in dynamically adjusting physical field parameters according to the microbial growth stages: during the lag phase, only a magnetic field is applied to promote spore germination; during the early logarithmic phase, intermittent ultrasound is applied to enhance cell membrane permeability; during the late logarithmic phase, continuous ultrasound combined with a high-intensity magnetic field is applied to drive metabolic flow towards product synthesis; and during the stationary phase, mild stimulation is restored to maintain cell activity. Real-time monitoring is achieved through an online viscometer, dissolved oxygen sensor, and near-infrared enzyme activity sensor, with the PLC controller automatically adjusting the field strength parameters to realize precise closed-loop control throughout the entire process.

[0017] In terms of equipment structure, this invention deeply integrates mechanical stirring and ultrasonic emission. The stirring box of the ultrasonic stirring device can rotate with the drive turntable and oscillate under the drive of the power component, so that the ultrasonic waves are dynamically directionally emitted within the reactor, completely eliminating ultrasonic dead zones. The stirring box is equipped with a cross-shaped plate and eight transducers, which, together with a wavelength acoustic matching plate, perform impedance matching to ensure efficient and uniform conduction of ultrasonic energy. The power supply component adopts a rotating conductive ring structure to solve the power supply problem of moving parts. The cooling device constructs an active liquid cooling cycle through heat pipes and semiconductor cooling chips to prevent local high temperatures from affecting bacterial activity. The sound-absorbing components on the outer wall of the tank absorb structural noise. The magnetic field generating device adopts a segmented coil design and integrates airflow cooling channels and shielding covers to ensure the strength and directionality of the magnetic field while preventing magnetic field leakage. Attached Figure Description

[0018] Figure 1 This is a flow chart of the microbial agent preparation process of the present invention; Figure 2 This is a control system framework diagram of the present invention; Figure 3 This is an isometric view of the intelligent multi-field coupled reactor structure of the present invention; Figure 4 This is an exploded view of the intelligent multi-field coupled reactor structure of the present invention; Figure 5 This is an isometric view of the magnetic field generating device of the present invention; Figure 6 This is an exploded view of the magnetic field generating device of the present invention; Figure 7 For the present invention Figure 6 Enlarged view of the structure at point A in the middle; Figure 8 This is an isometric view of the power supply component structure of the present invention; Figure 9 This is an isometric view of the power component structure of the present invention; Figure 10 For the present invention Figure 9 Enlarged view of the structure at point B in the middle; Figure 11 This is an isometric view of the cooling device structure of the present invention; Figure 12This is a cross-sectional view of the intelligent multi-field coupled reactor structure of the present invention; Figure 13 For the present invention Figure 12 Enlarged view of the structure at point C.

[0019] Figure Descriptions: 10. Tank body; 20. Ultrasonic stirring device; 21. Drive turntable; 22. Rotating tube; 23. Stirring box; 24. Ultrasonic generating component; 241. Cross-shaped plate; 242. Transducer; 243. Wavelength acoustic matching plate; 25. Power component; 251. Ring frame; 252. Electric telescopic cylinder; 253. First lifting ring; 254. Vertical plate; 255. Second lifting ring; 256. Support rod; 257. Drive rod; 26. Power supply component; 261. Bracket; 262. End cap; 263. Conductive stator; 2 64. Rotor conductive ring; 27. Silencing component; 271. Silencing cover plate; 272. Silencing groove; 30. Magnetic field generating device; 31. Insulating cover plate; 32. Through hole; 33. First airflow ring; 34. Second airflow ring; 35. Coil section; 36. Shielding cover; 40. Cooling device; 41. Circulating pump; 42. Liquid inlet ring; 43. Liquid outlet ring; 44. Heat conduction pipe; 45. Input pipe; 46. Output pipe; 47. Heat exchange component; 471. Heat exchange section; 472. Heat exchange fins; 473. Mounting cover; 474. Semiconductor refrigeration chip. Detailed Implementation

[0020] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention.

[0021] The embodiments of the present invention will now be described.

[0022] Please refer to the appendix for details. Figure 1 , 2As shown in Figure 3, in a preferred embodiment of the present invention, a process for preparing a high-efficiency microbial degradation liquefied cellulose composite inoculant includes the following steps: Step 1, inoculant activation and seed liquid preparation: After activating the cellulose-degrading inoculant, it is inoculated into a seed culture medium to obtain seed liquid 1; Step 2, inoculation and fermentation: Seed liquid 1 is inoculated into a fermentation culture medium and fermented in an intelligent multi-field coupling reactor; Step 3, multi-field coupling enhancement and online detection: During fermentation, an ultrasonic stirring device 20 applies an ultrasonic field, and a magnetic field generating device 30 applies a magnetic field to achieve synergistic enhancement, and fermentation parameters are detected in real time by an online viscometer, a fluorescence dissolved oxygen sensor, and a near-infrared enzyme activity sensor; Step 4, intelligent feedback control: Based on the online detection results of fermentation parameters, the controller automatically adjusts the ultrasonic intensity of the ultrasonic stirring device 20, the magnetic field intensity of the magnetic field generating device 30, and the fermentation process parameters; Step 5, inoculant post-treatment: After fermentation, the fermentation broth is collected and post-treated to obtain... The composite microbial agent is finished; the intelligent multi-field coupled reactor includes a tank 10 for carrying the microbial agent, an ultrasonic stirring device 20 installed inside the tank 10, a magnetic field generating device 30 installed on the outer wall of the tank 10, and an online viscometer, a fluorescence dissolved oxygen sensor, and a near-infrared enzyme activity sensor for detecting the microbial agent inside the tank 10. In step three, a staged multi-field enhancement strategy is adopted according to the fermentation stage: Lag phase: magnetic field strength of 50-100 mT is applied, no ultrasound is applied; Early logarithmic phase: ultrasonic strength of 20-30 kHz and magnetic field strength of 100-150 mT are applied; Late logarithmic phase: ultrasonic strength of 30-40 kHz and magnetic field strength of 150-200 mT are applied; Stationary phase: ultrasonic strength of 20-30 kHz and magnetic field strength of 80-120 mT are applied; the ultrasound in the early logarithmic phase is applied intermittently with a duty cycle of 30-50%; the ultrasound in the late logarithmic phase is applied continuously; the ultrasound in the stationary phase is applied intermittently with a duty cycle of 25-35%.

[0023] It should be noted that in this embodiment, Trichoderma reesei and Aspergillus niger were inoculated into PDA slant culture medium and cultured in a 30°C constant temperature incubator for 4 days to obtain activated slant. The bacterial strains from the activated slant were picked and inoculated onto seed culture medium, and incubated at 30℃ and 200 rpm for 36 hours to obtain the seed culture. The concentration of *Trichoderma reesei* seed culture was 2.5 × 10⁻⁶ using the plate count method. 8 CFU / mL, the concentration of Aspergillus niger seed culture was 3.0 × 10⁻⁶. 8 CFU / mL. Mix the two seed solutions at a 1:1 volume ratio and set aside. The tank 10 has a total volume of 50L and a filling coefficient of 70%. It is equipped with an ultrasonic stirrer 20, a magnetic field generator 30, an online viscometer, a fluorescence dissolved oxygen sensor, and a near-infrared enzyme activity sensor. It uses a PLC controller and features a touchscreen human-machine interface. Before use, the tank 10 is sterilized by emptying, then the fermentation medium is filled in, and then sterilized again. After sterilization, it is cooled to 30°C. The mixed seed culture was inoculated into the reactor at an 8% (v / v) inoculum rate. Fermentation parameters were set as follows: temperature 30℃, aeration rate 1.0 vvm, initial stirring speed 200 rpm, and fermentation cycle 36 hours.

[0024] Lag period (0-4h): Magnetic field strength of 80mT is applied by magnetic field generator 30, while ultrasonic stirring device 20 does not apply ultrasound. During this stage, the magnetic field acts on the spores, promoting spore germination and initial mycelial growth.

[0025] Early logarithmic phase (4-12h): Ultrasonic stirring device 20 applies ultrasonic intensity of 25kHz, using intermittent operation mode with a duty cycle of 40%; magnetic field generator 30 applies magnetic field intensity of 120mT. The intermittent stimulation of ultrasound enhances cell membrane permeability and promotes the transport of nutrients into the cell, while the magnetic field enhances intracellular synthesis and metabolism. The two work synergistically to promote rapid mycelial growth.

[0026] Late logarithmic phase (12-24h): Ultrasonic stirring device 20 applies ultrasonic intensity of 35kHz, operating in continuous mode; magnetic field generator 30 applies magnetic field intensity of 180mT. The cavitation effect generated by continuous ultrasound helps the intracellularly synthesized cellulase to be secreted outward, and the high-intensity magnetic field drives the metabolic flow in the direction of product synthesis, significantly increasing cellulase yield.

[0027] Stable period (24-36h): Ultrasonic stirring device 20 applies ultrasonic intensity of 25kHz, resuming intermittent operation mode with a duty cycle of 30%; magnetic field generator 30 applies magnetic field intensity of 100mT. Mild physical field stimulation maintains cell metabolic activity, delays aging and autolysis, and prolongs the enzyme production cycle.

[0028] The core of intelligent feedback control lies in the controller's built-in multi-parameter coupled control model. This model pre-sets multiple control rules matching the fermentation stage. First, the controller comprehensively determines the growth stage of the microorganism based on data collected by online sensors and the fermentation time. Then, the model compares key parameters such as viscosity, dissolved oxygen, and enzyme activity at the current moment with preset optimization target thresholds. When a parameter deviates from the threshold range or its rate of change exceeds the preset interval, the controller automatically calculates and outputs new execution instructions according to the preset control rule priority. This dynamically adjusts one or more parameters among ultrasonic intensity, ultrasonic operating mode, magnetic field strength, stirring speed, and aeration rate until the fermentation parameters return to the target range, thereby achieving closed-loop precise control of the fermentation process.

[0029] Intelligent feedback control record: At 18 hours: the near-infrared enzyme activity sensor detected that the enzyme activity growth rate had dropped to 4.2% / h, below the set threshold of 5% / h. The PLC controller automatically increased the ultrasonic intensity from 35kHz to 42kHz, and maintained it for 30 minutes before reverting to normal. After adjustment, the enzyme activity growth rate recovered to 6.8% / h.

[0030] At 22 hours: the dissolved oxygen sensor detected that the dissolved oxygen concentration had dropped to 18%, below the set threshold of 20%. The PLC controller automatically increased the stirring speed from 200 rpm to 220 rpm, the aeration rate from 1.0 vvm to 1.2 vvm, and simultaneously adjusted the ultrasonic duty cycle from continuous mode to 50% intermittent mode. After the adjustment, the dissolved oxygen gradually recovered to 28%.

[0031] At 28 hours: the viscometer detected a viscosity increase rate of 12% / h, exceeding the set threshold of 10% / h. The PLC controller automatically reduced the ultrasonic intensity from 35kHz to 28kHz, and restored it after 1 hour to prevent excessive ultrasonic waves from damaging the bacteria.

[0032] Fermentation was completed after 36 hours, and the fermentation broth was collected. The wet bacterial cells were separated using a tubular centrifuge. Based on the wet weight of the bacterial cells, 5% skim milk powder and 2% trehalose were added as preservatives and mixed thoroughly. The mixture was then dried using a vacuum freeze dryer for 24 hours to obtain the final compound bacterial agent.

[0033] The test results showed that the viable count of the compound microbial agent was 8.5 × 10⁻⁶. 10 CFU / g, filter paper enzyme activity 1200 IU / g, β-glucosidase activity 450 IU / g.

[0034] The obtained compound microbial agent was added to the degradation system of corn straw pretreated with 5% alkali at an inoculation rate of 1% (w / w). The mixture was treated at 50℃ and pH 5.5 for 48 hours. The reducing sugar yield was 32.5 g / L and the cellulose degradation rate was 87.3% as determined by the DNS method.

[0035] Comparative Example 1: Conventional Fermentation Process Fermentation was carried out in a conventional fermenter without the application of ultrasound or magnetic fields; only temperature, stirring, and aeration were controlled. Fermentation was completed after 72 hours, and samples were taken for analysis: the cell density was 22 g / L, and the filter paper enzyme activity was 9.5 IU / mL. The resulting inoculum had a viable count of 4.2 × 10⁻⁶. 10 CFU / g, filter paper enzyme activity 680 IU / g. Cellulose degradation experiment: reducing sugar yield 18.6 g / L, degradation rate 52.8% after 48 h.

[0036] Comparative Example 2: Single Ultrasonic Field Enhancement Fermentation was performed using only ultrasound, without applying a magnetic field. Fermentation lasted 48 hours, with a cell density of 28 g / L and filter paper enzyme activity of 12.8 IU / mL. The resulting inoculum contained 6.1 × 10⁻⁶ viable cells. 10 CFU / g, filter paper enzyme activity 890 IU / g. Cellulose degradation experiment after 48 hours: reducing sugar yield 24.3 g / L, degradation rate 67.5%.

[0037] Comparative Example 3: Enhancement by a Single Magnetic Field Fermentation was carried out using only a magnetic field, without ultrasound. Fermentation lasted 52 hours, with a cell density of 26 g / L and filter paper enzyme activity of 11.2 IU / mL. The resulting inoculum had a viable count of 5.5 × 10⁻⁶ cells / mL. 10 CFU / g, filter paper enzyme activity 780 IU / g. Cellulose degradation experiment: reducing sugar yield 21.8 g / L after 48 h, degradation rate 61.2%.

[0038] Please refer to the appendix for details. Figure 4 , 5As shown in Figures 6, 7, 8, 9, 10, and 12, in another preferred embodiment of the present invention, the ultrasonic stirring device 20 includes a drive turntable 21 rotatably connected to the top of the tank 10, a rotating tube 22 passing through the drive turntable 21, a plurality of stirring boxes 23 with one end hinged to the outer wall of the rotating tube 22 and arranged in a ring array, an ultrasonic generating component 24 disposed in the stirring box 23, and a power component 25 disposed on the top of the drive turntable 21 for simultaneously driving the plurality of stirring boxes 23 to swing. The ultrasonic generating component 24 includes a cross-shaped plate 241 disposed in the stirring box 23, symmetrically disposed on the cross-shaped plate 241. The device comprises eight transducers 242 at its four ends, and a wavelength acoustic matching plate 243 with one end abutting against the transducers 242 and the other end abutting against the inner wall of the mixing box 23. The power component 25 includes an annular frame 251 at the top of the mixing box 23, an electric telescopic cylinder 252 at the top of the drive turntable 21 with its actuating end penetrating the drive turntable 21, a first lifting ring 253 at the actuating end of the electric telescopic cylinder 252 and sleeved on the outer wall of the rotating tube 22, multiple vertical plates 254 with their top ends connected to the bottom of the first lifting ring 253, and plates with their top ends connected to the bottom ends of the vertical plates 254 and sleeved on the outer wall of the rotating tube 22. The second lifting ring 255 includes a plurality of support rods 256 arranged in a ring array on the outer wall of the second lifting ring 255, and a drive rod 257 with one end connected to the end of the support rod 256 and the other end slidably connected to the inner wall of the ring frame 251. It also includes a power supply component 26 located at the top of the tank body 10. The power supply component 26 includes an end cap 262 fixed to the top of the tank body 10 via a bracket 261, a conductive stator 263 located within the end cap 262, and a rotor conductive ring 264 located on the inner wall of the rotating tube 22 and in contact with the conductive stator 263. It also includes a noise reduction component 27 located on the outer wall of the tank body 10. The noise reduction component 27 includes a noise reduction cover plate 271 disposed on the outer wall of the tank body 10. The noise reduction cover plate 271 has a plurality of noise reduction grooves 272 on the side near the tank body 10. The magnetic field generating device 30 includes an insulating cover plate 31 disposed on the outer wall of the noise reduction cover plate 271, a plurality of through holes 32 passing through the insulating cover plate 31, a first airflow ring 33 and a second airflow ring 34 respectively disposed on the top and bottom of the insulating cover plate 31 and communicating with the through holes 32, a plurality of coil segments 35 disposed on the outer wall of the insulating cover plate 31, and a shielding cover 36 disposed on the outer wall of the tank body 10 and covering the coil segments 35.

[0039] It should be noted that, in this embodiment, when the ultrasonic stirring device 20 is working, the motor drives the drive turntable 21 to rotate, and the drive turntable 21, the rotating tube 22, and the stirring box 23 rotate together. The ultrasonic generating component 24 in the stirring box 23 can generate ultrasonic waves, which are transmitted to the bacterial solution through the stirring box 23. The actuator of the power component 25 can drive the stirring box 23 to rotate around the hinge point between the stirring box 23 and the rotating tube 22, so as to adjust the emission direction of the ultrasonic waves. Furthermore, when the drive turntable 21 rotates, the motor actuator in the mounting box at the top of the tank 10 drives the drive gear to rotate, and the drive gear meshes with the outer wall of the drive turntable 21 to drive the drive turntable 21 to rotate. Furthermore, when the ultrasonic generating component 24 is working, the ultrasonic generator installed on the tank 10 supplies power to the transducer 242 through wires and power supply component 26, and the radiating surface of the transducer 242 transmits ultrasonic waves to the bacterial agent through the wavelength acoustic matching plate 243 and the stirring box 23. The wavelength acoustic matching plate 243 is made of a composite material of epoxy resin and tungsten powder. By adjusting the mass fraction of tungsten powder, its acoustic impedance reaches 6.5 MRayl. Furthermore, when the power unit 25 is working, the actuator of the electric telescopic cylinder 252 drives the first lifting ring 253, the vertical plate 254, the second lifting ring 255, the support rod 256, and the drive rod 257 to rise or fall. The drive rod 257 slides within the annular frame 251 and drives the mixing box 23 to swing through the annular frame 251. Furthermore, the end of the mixing box 23 is connected to the rotating tube 22 via a flexible hose. When the power supply component 26 is working, the conductive stator 263 adopts a multi-point cluster brush design, consisting of multiple precious metal alloy brush filaments, evenly distributed around the rotor conductive ring 264, with a contact pressure of 8-12g. The rotor conductive ring 264 uses a beryllium copper alloy substrate with a hard gold plating treatment on the surface and is provided with a V-shaped annular groove to increase the contact area. Furthermore, when the silencing component 27 is in operation, the silencing cover 271 is made of 304 stainless steel with a thickness of 2mm, and the silencing groove 272 is wedge-shaped with a depth of 12mm, a width of 8mm, and a spacing of 10mm. The silencing groove 272 is filled with silicone rubber-based composite sound-absorbing material to eliminate sound pressure and effectively improve the uniformity of the ultrasonic field; Furthermore, when the magnetic field generating device 30 is working, each coil segment 35 is powered by an independent constant current source, and the output current of the constant current source is adjustable from 0-10A with an accuracy of ±0.2%. The PLC controller sets the current value of each coil segment 35 according to the fermentation stage, and adjusts it in real time through the Hall sensor installed in the tank 10 to achieve precise control of the magnetic field strength; The input end of the first airflow ring 33 can be connected to the air source system. The airflow enters through the first airflow ring 33, undergoes heat exchange through the through hole 32, and is discharged through the second airflow ring 34 to cool the coil section 35.

[0040] Please refer to the appendix for details. Figure 8 , 11 As shown in Figures 12 and 13, in another preferred embodiment of the present invention, a cooling device 40 is further included. The cooling device 40 includes a circulating pump 41 disposed at the top of the drive turntable 21, an inlet ring 42 and a drain ring 43 sequentially disposed from top to bottom on the inner wall of the rotating tube 22, and a heat-conducting pipe 44 embedded in the cross-shaped plate 241; the inlet end of the heat-conducting pipe 44 is connected to the inlet ring 42, and the drain end of the heat-conducting pipe 44 is connected to the drain ring 43. 2. The pump is connected to the output end of the circulating pump 41 via the input pipe 45. The drain ring 43 is connected to the input end of the circulating pump 41 via the output pipe 46. The pump also includes a heat exchange component 47. The heat exchange component 47 includes a heat exchange section 471 on the output pipe 46, a plurality of heat exchange fins 472 on the heat exchange section 471, a mounting cover 473 on the outer wall of the end cover 262, and a plurality of semiconductor cooling chips 474 inside the mounting cover 473.

[0041] It should be noted that, in this embodiment, when the cooling device 40 is working, the circulating pump 41 drives the coolant to flow in a closed circulation system consisting of an input pipe 45, an inlet ring 42, a heat conduction pipe 44, a drain ring 43, and an output pipe 46: the coolant enters the inlet ring 42 through the input pipe 45, then flows to each heat conduction pipe 44, absorbs the heat generated by the transducer 242, and then flows into the drain ring 43, enters the heat exchange section 471 through the output pipe 46, is cooled by the semiconductor cooling chip 474, and then returns to the circulating pump 41; Furthermore, based on the feedback from the temperature sensor corresponding to the transducer 242, the PLC controller automatically adjusts the power of the semiconductor cooling chip 474 and the speed of the circulating pump 41 to keep the temperature of the transducer 242 always below the set value. The heat exchange fin structure 472 can improve heat exchange efficiency.

[0042] Although embodiments of the present invention have been shown and described, these specific embodiments are merely explanations of the invention and are not intended to limit it. The specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. After reading this specification, those skilled in the art may make modifications, substitutions, and variations to the embodiments as needed without departing from the principles and spirit of the invention, but such modifications, substitutions, and variations are protected by patent law as long as they are within the scope of the claims of the present invention.

Claims

1. A process for preparing a highly efficient microbial degradation liquefied cellulose composite inoculant, characterized in that... This includes the following steps: Step 1: Activation of the strain and preparation of seed culture. After activating the cellulose-degrading strain, it is inoculated into the seed culture medium for cultivation to obtain seed culture one. Step 2, Inoculation and Fermentation: Inoculate the seed liquid into the fermentation medium and carry out fermentation culture in an intelligent multi-field coupled reactor. Step 3: Multi-field coupling enhancement and online detection. During the fermentation process, the ultrasonic stirring device (20) applies an ultrasonic field, and the magnetic field generating device (30) applies a magnetic field to achieve synergistic enhancement. Fermentation parameters are detected in real time by an online viscometer, a fluorescence dissolved oxygen sensor, and a near-infrared enzyme activity sensor. Step 4: Intelligent feedback control. Based on the online detection results of fermentation parameters, the controller automatically adjusts the ultrasonic intensity of the ultrasonic stirring device (20), the magnetic field intensity of the magnetic field generator (30), and the fermentation process parameters. Step 5: Post-treatment of microbial agent. After fermentation, the fermentation broth is collected and post-treated to obtain the compound microbial agent product. The intelligent multi-field coupled reactor includes a tank (10) for carrying bacterial agents, an ultrasonic stirring device (20) disposed in the tank (10), a magnetic field generating device (30) disposed on the outer wall of the tank (10), and an online viscometer, a fluorescent dissolved oxygen sensor and a near-infrared enzyme activity sensor for detecting bacterial agents in the tank (10).

2. The preparation process of a high-efficiency microbial degradation liquefied cellulose composite inoculant according to claim 1, characterized in that, In step three, a staged multi-field enhancement strategy is adopted according to the fermentation stage: Delay period: Apply a magnetic field strength of 50-100 mT, without applying ultrasound; Pre-log phase: Apply ultrasound at 20-30 kHz and magnetic field at 100-150 mT; Late logarithmic phase: Apply ultrasound at 30-40 kHz and magnetic field at 150-200 mT; Stable period: Apply ultrasonic intensity of 20-30kHz and magnetic field strength of 80-120mT; The ultrasound during the pre-log phase is applied intermittently with a duty cycle of 30-50%; the ultrasound during the post-log phase is applied continuously; and the ultrasound during the stationary phase is applied intermittently with a duty cycle of 25-35%.

3. The preparation process of a high-efficiency microbial degradation liquefied cellulose composite inoculant according to claim 1, characterized in that, The ultrasonic stirring device (20) includes a drive turntable (21) rotatably connected to the top of the tank (10), a rotating tube (22) passing through the drive turntable (21), a plurality of stirring boxes (23) with one end hinged to the outer wall of the rotating tube (22) and arranged in a ring array, an ultrasonic generating component (24) disposed in the stirring box (23), and a power component (25) disposed on the top of the drive turntable (21) for simultaneously driving the plurality of stirring boxes (23) to swing.

4. The preparation process of a high-efficiency microbial degradation liquefied cellulose composite inoculant according to claim 3, characterized in that, The ultrasonic generating component (24) includes a cross-shaped plate (241) disposed in the stirring box (23), eight transducers (242) symmetrically disposed at the four ends of the cross-shaped plate (241), and a wavelength acoustic matching plate (243) with one end abutting against the transducer (242) and the other end abutting against the inner wall of the stirring box (23).

5. The preparation process of a high-efficiency microbial degradation liquefied cellulose composite agent according to claim 3, characterized in that, The power unit (25) includes an annular frame (251) on top of the mixing box (23), an electric telescopic cylinder (252) on top of the drive turntable (21) with its execution end penetrating the drive turntable (21), a first lifting ring (253) on the execution end of the electric telescopic cylinder (252) and sleeved on the outer wall of the rotating tube (22), a plurality of vertical plates (254) with their top ends connected to the bottom of the first lifting ring (253), a second lifting ring (255) with its top ends connected to the bottom of the vertical plates (254) and sleeved on the outer wall of the rotating tube (22), a plurality of support rods (256) arranged in a ring array on the outer wall of the second lifting ring (255), and a drive rod (257) with one end connected to the end of the support rod (256) and the other end slidably connected to the inner wall of the annular frame (251).

6. The preparation process of a high-efficiency microbial degradation liquefied cellulose composite inoculant according to claim 4, characterized in that, It also includes a power supply component (26) located on the top of the tank (10). The power supply component (26) includes an end cap (262) fixed to the top of the tank (10) by a bracket (261), a conductive stator (263) located in the end cap (262), and a rotor conductive ring (264) located on the inner wall of the rotating tube (22) and in contact with the conductive stator (263).

7. The preparation process of a highly efficient microbial degradation liquefied cellulose composite inoculant according to claim 6, characterized in that, It also includes a cooling device (40), which includes a circulating pump (41) located on the top of the drive turntable (21), an inlet ring (42) and a drain ring (43) arranged sequentially from top to bottom on the inner wall of the rotating tube (22), and a heat-conducting tube (44) embedded in the cross-shaped plate (241). The inlet end of the heat pipe (44) is connected to the inlet ring (42), the outlet end of the heat pipe (44) is connected to the outlet ring (43), the inlet ring (42) is connected to the outlet end of the circulation pump (41) through the input pipe (45), and the outlet ring (43) is connected to the input end of the circulation pump (41) through the output pipe (46).

8. The preparation process of a high-efficiency microbial degradation liquefied cellulose composite agent according to claim 7, characterized in that, It also includes a heat exchange component (47), which includes a heat exchange section (471) disposed on the output pipe (46), a plurality of heat exchange fins (472) disposed on the heat exchange section (471), a mounting cover (473) disposed on the outer wall of the end cover (262), and a plurality of semiconductor cooling chips (474) disposed in the mounting cover (473).

9. The preparation process of a high-efficiency microbial degradation liquefied cellulose composite agent according to claim 3, characterized in that, It also includes a sound-absorbing component (27) disposed on the outer wall of the tank (10). The sound-absorbing component (27) includes a sound-absorbing cover plate (271) disposed on the outer wall of the tank (10). The sound-absorbing cover plate (271) has a plurality of sound-absorbing grooves (272) on the side near the tank (10).

10. The preparation process of a high-efficiency microbial degradation liquefied cellulose composite inoculant according to claim 9, characterized in that, The magnetic field generating device (30) includes an insulating cover plate (31) disposed on the outer wall of the silencing cover plate (271), a plurality of through holes (32) passing through the insulating cover plate (31), a first airflow ring (33) and a second airflow ring (34) respectively disposed on the top and bottom of the insulating cover plate (31 and communicating with the through holes (32), a plurality of coil segments (35) disposed on the outer wall of the insulating cover plate (31), and a shielding cover (36) disposed on the outer wall of the tank (10) and covering the coil segments (35).

Images