Solid state bioreactor and methods of use thereof

Abstract

The application discloses a kind of solid-state biological reaction device and its use method, it is related to solid-state fermentation technical field, device includes solid-state bioreactor and multifunctional fermentation cabin;The reactor main cavity is divided into upper fermentation cavity and lower air chamber by sieve plate, sieve plate supports material, outside temperature control component is equipped, lower air chamber is equipped with air distribution component and drain pipe;Top cover closes upper opening, is equipped with exhaust pipe and at least four high-density resistivity probes of diameter equidistant arrangement and connection high-density resistivity detection system, its lower part is inserted into material surface layer, and relevant surface is equipped with insulating layer;Multifunctional fermentation cabin inner cavity is divided into upper, middle and lower three cavities, lower cavity is placed reactor;Middle cavity is equipped with hand hole with rubber glove, opening adjustable roller blind and opposite air purification unit, so that the upper opening of reactor is located in the airflow area between the two;Upper cavity is equipped with lifting equipment for lifting top cover and heating component, and there is external connection port.The use method is the use instruction of solid-state biological reaction device.

Description

Technical Field

[0001] This invention relates to the field of solid-state fermentation technology, and in particular to a solid-state bioreactor and its method of use. Background Technology

[0002] Solid-state fermentation is widely used in the production of alcoholic beverages, food, feed, and biopesticides. Unlike liquid fermentation, the moisture content inside the solid-state fermentation bed exhibits a non-uniform gradient distribution and changes dynamically over time. Without real-time data acquisition, localized overheating or water loss can easily occur, affecting fermentation quality. However, existing solid-state bioreactors lack the capability to characterize the spatial distribution of moisture content within the bed online.

[0003] Furthermore, existing solid-state reactors (such as packed beds) often lack reliable local sterile environments during loading, inoculation, and sampling, resulting in a high risk of contamination. Traditional methods rely on large sterile rooms, which suffer from drawbacks such as low temperature control accuracy, high energy consumption, and easy contamination due to personnel entering and exiting the rooms. Summary of the Invention

[0004] The purpose of this invention is to provide a solid-state bioreactor and its usage method to solve the problems existing in the prior art, realize online detection of bed moisture content, construct a sterile and precisely temperature-controlled operating environment, reduce the risk of contamination, and improve the controllability of the fermentation process.

[0005] To achieve the above objectives, the present invention provides the following solution: This invention provides a solid-state bioreactor, comprising: a solid-state bioreactor, which includes a reactor body; the inner cavity of the reactor body is divided into an upper fermentation chamber and a lower gas chamber by a sieve plate, the upper fermentation chamber being located above the lower gas chamber; the sieve plate being used to support fermentation materials; a temperature control component corresponding to the position of the upper fermentation chamber is provided on the outer wall of the reactor body; an air distribution component for introducing sterile air is provided in the lower gas chamber; a top cover is sealed at the upper opening of the reactor body; an exhaust pipe and at least four high-density resistivity probes are fixed on the top cover; a drain pipe is provided at the bottom of the lower gas chamber; the top cover is coaxially arranged with the reactor body; the high-density resistivity probes are arranged at equal intervals along the diameter direction passing through the center of the top cover; the lower part of each high-density resistivity probe is used to insert into the surface layer of the fermentation materials at equal depths; and each high-density resistivity probe... A resistivity probe is connected to a high-density resistivity detection system; an insulating layer is provided on the surfaces of the upper fermentation chamber, the top cover, and the sieve plate facing the fermentation material; a multi-functional fermentation chamber has an internal cavity divided into an interconnected upper cavity, a middle cavity, and a lower cavity; the lower cavity is used to place the solid bioreactor, and the bottom of the lower cavity has a lower opening; multiple hand holes are provided on the side wall of the middle cavity, and rubber gloves are provided on the hand holes; a roller shutter with adjustable opening is provided on the side wall of the middle cavity and is connected to the outside; an air purification unit is provided on the side wall of the middle cavity opposite to the roller shutter; in the vertical direction, the upper opening of the reactor body is located in the horizontal airflow area between the air purification unit and the roller shutter; the upper cavity is provided with a lifting device and a heating component; the lifting device is used to lift the top cover, and the upper cavity has an external connection port connected to the outside.

[0006] Preferably, the high-density resistivity probe includes a fixed external connector, a probe telescopic cylinder, a probe rod, and an energized connector; the fixed external connector can be fixedly connected to the top cover, and the upper end of the fixed external connector is connected to the high-density resistivity detection system via a wire; the probe telescopic cylinder can be fixed to the top cover inside the upper fermentation chamber, and the lower end of the fixed external connector is located inside the inner cavity of the probe telescopic cylinder; the probe telescopic cylinder is made of a non-conductive material; the upper end of the probe rod slides vertically within the inner cavity of the probe telescopic cylinder, and the lower end of the probe rod is used to insert into the surface layer of the fermentation material; the energized connector is located within the inner cavity of the probe telescopic cylinder between the fixed external connector and the probe rod, and the energized connector is used to energize the fixed external connector and the probe rod.

[0007] Preferably, the lower end of each of the probe rods is fixed to the lower positioning plate; the lower positioning plate is located inside the upper fermentation chamber, and in the vertical direction, the lower positioning plate is higher than the lower end of the probe rod.

[0008] Preferably, the top cover is provided with a plurality of guide components, each guide component having a guide rod that extends and retracts in the vertical direction, and the lower positioning plate is fixedly connected to the lower end of each guide rod.

[0009] Preferably, the energized connector is a metal spring, and the metal spring is in a compressed state located between the fixed external connector and the probe rod.

[0010] Preferably, the power-conducting connector is a metal braided strip, and the lower positioning plate is weighted.

[0011] Preferably, each of the fixed external components is fixed to the upper positioning plate, and the top cover has a through mounting hole, and the upper positioning plate is fixedly inserted into the mounting hole.

[0012] Preferably, the top cover is further provided with multiple multi-point temperature measuring probes and / or multiple imaging components; each of the multi-point temperature measuring probes is distributed in a V-shape around the axis of the top cover, or each of the multi-point temperature measuring probes is distributed in a straight line along the diameter direction passing through the center of the top cover; and the distance between the axis of each multi-point temperature measuring probe and the axis of the top cover is different; the lower end of each multi-point temperature measuring probe is inserted into the fermentation material; each of the imaging components is distributed in a V-shape around the axis of the top cover, or each of the imaging components is distributed in a straight line along the diameter direction passing through the center of the top cover; and the distance between the axis of each imaging component and the axis of the top cover is different; the imaging component includes a pointed blind tube and an endoscope, the lower end of the pointed blind tube is inserted into the fermentation material, and the endoscope is placed inside the pointed blind tube.

[0013] Preferably, the sieve plate has a groove in the center, and stainless steel wires are arranged in the groove; an insulating terminal is provided on the side wall of the lower air chamber, one end of the stainless steel wire is connected to the insulating terminal through a lead wire, and the insulating terminal is connected to the dedicated terminal of the high-density resistivity detection system through a wire.

[0014] The present invention also provides a method of using the solid-state bioreactor based on any one of the above claims, comprising the following steps: S1, The solid bioreactor is placed in the lower chamber of the multifunctional fermentation chamber, and the interior of the solid bioreactor is disinfected; S2, turn on the air purification unit, turn off the heating component, open the roller shutter and load the sterilized and cooled solid fermentation substrate onto the sieve plate, pour in the inoculum, reduce the opening of the roller shutter, and use the rubber gloves to hold the sterile tool to stir the solid fermentation substrate mixed with the inoculum evenly. S3, close the top cover, turn on the temperature control component, and introduce sterile air regulated by temperature and humidity into the lower air chamber through the air distribution component at a predetermined flow rate; S4, turn off the air purification unit and the roller shutter, start the heating component to control the temperature of the inner cavity of the multifunctional fermentation chamber, and start fermentation; and use the host computer that is communicatively connected to the high-density resistivity detection system to monitor it.

[0015] The present invention achieves the following technical effects compared to the prior art: The solid-state bioreactor provided by this invention features at least four high-density resistivity probes fixed on the top cover, with each probe evenly spaced along a diameter passing through the center of the top cover. The lower part of each probe is used to insert into the surface of the fermentation material at equal depth and is connected to a high-density resistivity detection system. Combined with insulating layers on the fermentation chamber, top cover, and sieve plate facing the fermentation material, a complete resistivity detection circuit is formed. This allows for real-time acquisition of resistivity data at different locations within the material bed to invert the spatial distribution of moisture content, achieving visualized monitoring of the fermentation process. Furthermore, by setting up a multifunctional fermentation chamber with an upper, middle, and lower cavity, the solid-state bioreactor is placed in the lower cavity. Utilizing the opposing air purification units on the sidewall of the middle cavity and the adjustable-opening roller shutter, the upper opening of the reactor body is located within the lateral airflow area between the two cavities, forming a clean air protective layer during operation. Simultaneously, operators can work in a closed environment through rubber gloves provided with handholes. This eliminates the need for a large sterile room, allowing for the construction of a reliable local sterile operating environment within the middle cavity, significantly reducing the risk of contamination and improving temperature control accuracy and energy efficiency.

[0016] This invention also provides a method for using a solid-state bioreactor. Through the coordinated operation of the solid-state bioreactor and its multifunctional fermentation chamber components, aseptic and precise control of the entire solid-state fermentation process and real-time monitoring of the fermentation status are achieved. In the initial stage, the solid-state bioreactor is disinfected. During the inoculation stage, the air purification unit is activated, and rubber gloves are used to create a reliable localized aseptic environment, significantly reducing the risk of contamination. During the fermentation stage, the temperature control and heating components are activated simultaneously, combined with the aeration components to introduce temperature- and humidity-controlled aseptic air, achieving precise control of the fermentation temperature and aeration environment. During fermentation, a high-density resistivity detection system linked to a host computer monitors the fermentation status in real time, effectively preventing localized overheating and water loss in the material bed, and ensuring stable fermentation quality. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0018] Figure 1 A schematic diagram of the overall structure of the solid-state bioreactor provided by the present invention; Figure 2 This is a schematic diagram of the overall structure of the solid bioreactor in the solid bioreactor device provided by the present invention. Figure 3 This is a front view of the multifunctional fermentation chamber in the solid-state bioreactor provided by the present invention; Figure 4 Rear view of the multifunctional fermentation chamber in the solid-state bioreactor provided by the present invention; Figure 5 A side view of the multifunctional fermentation chamber in the solid-state bioreactor provided by the present invention; Figure 6 This is a schematic diagram showing the arrangement of high-density resistivity probes on the top cover of the solid-state bioreactor provided by the present invention. Figure 7 This is a schematic diagram of the combined structure of the upper positioning plate, guide assembly, lower positioning plate, and various high-density resistivity probes in the solid-state bioreactor provided by the present invention. Figure 8 A schematic diagram of the structure of the solid-state bioreactor provided by the present invention, which uses metal braided straps to fix the connection between the external component and the probe rod; Figure 9 This is a schematic diagram showing the arrangement of multi-point temperature probes on the top cover in the solid-state bioreactor provided by the present invention. Figure 10 A schematic diagram showing the multi-point temperature probes arranged in a V-shape on the top cover of the solid-state bioreactor provided by the present invention; Figure 11 A schematic diagram showing the multi-point temperature probes arranged in a straight line on the top cover of the solid-state bioreactor provided by the present invention; Figure 12 A schematic diagram showing the arrangement of the pointed blind tubes on the top cover in the solid-state bioreactor provided by the present invention; Figure 13 A schematic diagram showing the V-shaped arrangement of pointed blind tubes on the top cover in the solid-state bioreactor provided by the present invention; Figure 14 A schematic diagram showing the pointed blind tubes arranged in a straight line on the top cover in the solid bioreactor provided by the present invention; Figure 15 This is a schematic diagram showing the connection method of the three differential pressure gauges in the solid-state bioreactor provided by the present invention. Figure 16 The resistivity contour plot obtained after 72 hours of fermentation in Case 1; Figure 17 This is a contour map of the temperature field distribution obtained after 72 hours of fermentation in Case 1. Figure 18The temperature change trend at points 1 to 4 during the fermentation process in Case 1; Figure 19 These are images taken using an endoscope and a pointed blind tube to examine the interior of the fermentation material after 48 hours of fermentation in Case 2. Figure 20 This is a cross-sectional view of the cylindrical fermentation material after fermentation in Case 2. Figure 21 This is a graph showing the trend of pressure difference and weight changes during the entire 120-hour fermentation process in Case 3.

[0019] In the picture: 1-Reactor body; 2-Top cover; 3-Sieve plate; 4-Sealing ring; 5-Wire groove; 6-Lead wire; 7-Gas distribution pipe; 8-First temperature and humidity sensor; 9-Insulating terminal; 10-First pressure measuring tube; 11-Drain pipe; 12-Weight sensor; 13-Outrigger; 14-Jacket; 15-Inlet pipe; 16-Outlet pipe; 17-Exhaust pipe; 18-Second temperature and humidity sensor; 19-Multi-core cable; 20-Connecting connector; 21-High-density resistivity probe; 22-Guide rod; 23-Lower positioning plate; 24-Multifunctional fermentation chamber; 25-Sterilizing filter; 26-Enclosure panel; 27-Air supply fan; 28-Heating component; 29-Hanger; 30-Handhole; 31-Castwheel; 32-Retractable foot support; 33-Multi-point temperature probe; 34-Pointed blind tube; 35-Roller shutter; 40-Upper positioning plate; 41-Fixed external component; 42-Metal spring; 43-Probe rod; 44-Probe telescopic cylinder; 45-First cleaning hole; 46-Second pressure measuring tube; 47-Guide cylinder; 48-Second cleaning hole; 49-Counterweight; 50-Metal braided belt. 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. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0021] The purpose of this invention is to provide a solid-state bioreactor and its usage method to solve the problems existing in the prior art, realize online detection of the moisture content of the material bed, construct a sterile and precisely temperature-controlled operating environment, reduce the risk of contamination, and improve the controllability of the fermentation process.

[0022] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0023] Example 1 This embodiment provides a solid-state bioreactor, such as Figures 1-15 As shown, it includes: A solid-state bioreactor includes a reactor body 1. The inner cavity of the reactor body 1 is divided into an upper fermentation chamber and a lower gas chamber by a sieve plate 3. The upper fermentation chamber is located above the lower gas chamber, and the sieve plate 3 is used to support the fermentation material. A temperature control component corresponding to the position of the upper fermentation chamber is provided on the outer wall of the reactor body 1. An air distribution component for introducing sterile air is provided in the lower gas chamber. A top cover 2 is closed at the upper opening of the reactor body 1. An exhaust pipe 17 and at least four high-density resistivity probes 21 are fixed on the top cover 2 (the number can be reasonably selected, such as 24, 30, or 60, to obtain a high-density resistivity cloud map). A drain pipe 11 is provided at the bottom of the lower gas chamber. The top cover 2 is coaxially arranged with the reactor body 1, and each high-density resistivity probe 21 passes through the top cover 1. The high-density resistivity probes 21 are arranged at equal intervals along the diameter of the center of the top cover 2 (i.e., the diameter of the top cover 2 at the center of the circle passing through the upper surface of the top cover 2 from a top view). The lower part of each high-density resistivity probe 21 is used to insert into the surface of the fermentation material at the same depth. Each high-density resistivity probe 21 is connected to the high-density resistivity detection system (Chongqing Benteng WGMD-30 type). The detection signal of the high-density resistivity detection system is input to the host computer for inversion processing (using 2DRES software for inversion processing) to obtain the resistivity cloud map of the cross section. Each high-density resistivity probe 21 is connected to the high-density resistivity detection system through a multi-core cable 19. Insulation layers are provided on the surfaces of the upper fermentation chamber, the top cover 2, and the sieve plate 3 that face the fermentation material. The multifunctional fermentation chamber 24 has an internal cavity divided into an upper cavity, a middle cavity, and a lower cavity that are interconnected. The lower cavity is used to place the solid bioreactor, and the bottom of the lower cavity has a lower opening. The middle cavity has multiple hand holes 30 on its side wall, and rubber gloves are provided on the hand holes 30. The middle cavity side wall is also provided with a roller shutter 35 that is connected to the outside and has an adjustable opening. An air purification unit is provided on the middle cavity side wall opposite to the roller shutter 35. In the vertical direction, the upper opening of the reactor body 1 is located in the horizontal airflow area between the air purification unit and the roller shutter 35 (that is, in the vertical direction, the upper opening of the reactor body 1 is not lower than the lower edge of the air outlet of the air purification unit). The upper cavity is provided with a lifting device and a heating component 28. The lifting device is used to lift the top cover 2, and the upper cavity has an external connection port that connects to the outside.

[0024] By fixing at least four high-density resistivity probes 21 on the top cover 2, and arranging them at equal intervals along the diameter passing through the center of the top cover 2, the lower part of each probe is used to insert into the surface of the fermentation material at the same depth and is connected to the high-density resistivity detection system. Combined with the insulating layers on the fermentation chamber, the top cover 2, and the sieve plate 3 facing the fermentation material, a complete resistivity detection circuit is formed. This allows for real-time acquisition of resistivity data at different locations inside the material bed to invert the spatial distribution of moisture content, achieving visualized monitoring of the fermentation process. Furthermore, by setting internal... The multifunctional fermentation chamber 24 is divided into an upper chamber, a middle chamber, and a lower chamber. The solid bioreactor is placed in the lower chamber. The air purification units on the side wall of the middle chamber and the adjustable roller shutter 35 make the upper opening of the reactor body 1 located in the horizontal airflow area between the two, forming a clean air protection layer during operation. At the same time, the operator can work in a closed environment through the rubber gloves provided on the hand hole 30. A reliable local sterile operating environment can be built in the middle chamber without relying on a large sterile room, which significantly reduces the risk of contamination and improves temperature control accuracy and energy efficiency.

[0025] The following are the relevant settings for reactor body 1: Specifically, the reactor body 1 is cylindrical; the reactor body 1 is provided with an inner flange ring for receiving the sieve plate 3, which provides support for the sieve plate 3; at the same time, the sealing ring 4 at the lower edge of the sieve plate 3 is used to achieve sealing, preventing airflow from flowing through the gap between the sieve plate 3 and the inner flange ring.

[0026] Specifically, the top cover 2 is connected to the upper opening of the reactor body 1 via a flange.

[0027] Specifically, when high-density resistivity probes 21 are arranged on the top cover 2, the reactor body 1, the top cover 2, and the sieve plate 3 should all be made of polymer materials with good insulation properties (including but not limited to polypropylene, polytetrafluoroethylene, and polycarbonate), or made of metal materials, and a layer of polymer material with good insulation properties should be tightly attached to the surface facing the material.

[0028] The following are the settings instructions for each parameter monitoring component: Specifically, the setup instructions for the high-density resistivity probe 21 (which is used to detect moisture content using existing technology, and its working principle will not be elaborated further) are as follows: Among the optional solutions in this embodiment, the more preferred one is as follows: Figure 2As shown, a groove 5 is provided in the center of the sieve plate 3, and a stainless steel wire is arranged in the groove 5. An insulating terminal 9 is provided on the side wall of the lower air chamber. One end of the stainless steel wire is connected to the insulating terminal 9 through a lead wire 6 (one end of the lead wire 6 passes through the perforation in the center of the sieve plate 3 and connects to the insulating terminal 9 at the bottom). The insulating terminal 9 is connected to a dedicated terminal of the high-density resistivity detection system through a wire. With this configuration, the three-electrode method can be used to assist in the detection of high-density resistivity.

[0029] Among the optional solutions in this embodiment, the more preferred one is as follows: Figure 2 , Figure 7 and Figure 8 As shown, the high-density resistivity probe 21 includes a fixed external connector 41 (with a connecting joint 20), a probe telescopic cylinder 44 (with a first cleaning hole 45 communicating with the inner cavity), a probe rod 43, and an energized connector. The fixed external connector 41 can be fixedly connected to the top cover 2, and the upper end of the fixed external connector 41 is connected to the high-density resistivity detection system through a wire. The probe telescopic cylinder 44 can be fixed to the top cover 2 in the upper fermentation chamber, and the lower end of the fixed external connector 41 is located in the inner cavity of the probe telescopic cylinder 44. The probe telescopic cylinder 44 is made of a non-conductive material. The upper end of the probe rod 43 is slidably disposed in the inner cavity of the probe telescopic cylinder 44 in the vertical direction, and the lower end of the probe rod 43 is used to insert into the surface layer of the fermentation material. The energized connector is located in the inner cavity of the probe telescopic cylinder 44 between the fixed external connector 41 and the probe rod 43, and the energized connector is used to energize the fixed external connector 41 and the probe rod 43.

[0030] Specifically, both the fixed external connector 41 and the probe rod 43 are made of metal or other conductive materials, and are connected by a metal spring 42 or a flexible metal braided strip 50 to form a conductive path.

[0031] Among the optional solutions in this embodiment, the more preferred one is as follows: Figure 7 and Figure 8 As shown, the lower end of each probe rod 43 is fixed to the lower positioning plate 23; the lower positioning plate 23 is located in the upper fermentation chamber, and in the vertical direction, the lower positioning plate 23 is higher than the lower end of the probe rod 43 (the lower ends of each probe rod 43 are fixed to the lower positioning plate 23 with an interference fit at equal intervals).

[0032] Among the optional solutions in this embodiment, the more preferred one is as follows: Figure 7 and Figure 8 As shown, the top cover 2 is provided with multiple guide components (the guide components include guide cylinder 47 and guide rod 22, the guide cylinder 47 is fixed on the top cover 2, and the upper end of the guide rod 22 is slidably disposed in the guide cylinder 47; the guide cylinder 47 is provided with a second cleaning hole 48 communicating with its inner cavity), the guide components have guide rods 22 that extend and retract in the vertical direction, and the lower positioning plate 23 is fixedly connected to the lower end of each guide rod 22.

[0033] Specifically, the guide cylinder 47 and the guide rod 22 are fitted with a clearance. The depth of the internal space of the guide cylinder 47 is greater than the length of the thickened section at the head of the guide rod 22, so as to ensure that the head of the guide rod 22 can freely extend and retract up and down in the guide cylinder 47 under the drive of the lower positioning plate 23, and play a role in preventing the probe rod 43 from tilting.

[0034] Specifically, the fixed external component 41, probe telescopic cylinder 44, probe rod 43 and upper positioning plate 40 or top cover 2, as well as the guide cylinder 47, guide rod 22 and upper positioning plate 40 or top cover 2, can be detachable to facilitate effective sterilization of each component before the experiment; or they can be cleaned in conjunction with the first cleaning hole 45 and the second cleaning hole 48.

[0035] Specifically, the probe rod 43 and the inner wall of the probe telescopic cylinder 44 are fitted with a clearance, and the depth inside the probe telescopic cylinder 44 is left with a margin to ensure that the probe rod 43 can freely extend and retract in the vertical direction under the combined action of the metal spring 42 and the reaction force of the fermentation material.

[0036] Specifically, when the material bed shrinks during solid-state fermentation, the reduced surface height can lead to poor contact between the tip of the fixed resistivity probe and the material, affecting the high-density resistivity imaging results. In this case, an adaptive resistivity probe can be used instead of the fixed resistivity probe for detection. The first, and more preferred, option among the optional solutions in this embodiment is as follows: Figure 7 As shown, the power-conducting connector is a metal spring 42, and the metal spring 42 is in a compressed state (which can apply a vertically downward elastic force to the probe rod 43) and is located between the fixed external connector 41 and the probe rod 43.

[0037] The second, and more preferred, option among the optional solutions in this embodiment is as follows: Figure 8 As shown, the power-conducting connector is a metal braided strap 50, and the lower positioning plate 23 is reinforced (including an independently added counterweight 49 on the lower positioning plate 23 to increase the pressure of the lower positioning plate 23 carrying each probe rod 43 on the material bed, thereby ensuring that the tip of each probe rod 43 can make good contact with the material. When the spacing between the probe rods 43 is small and it is difficult to arrange the counterweight 49, the thickness of the lower positioning plate 23 can be increased to increase its own weight, achieving the same effect).

[0038] The first method utilizes the elastic force of each metal spring 42 to push each probe rod 43, and under the guidance of the guide rod 22, ensures that the lower end of each probe rod 43 is always stably connected to the material bed. In this case, additional weight can be added to the lower positioning plate 23 as needed. The specific method is as described above and will not be elaborated further. The second method utilizes the weight of the lower positioning plate 23 itself to drive each probe rod 43 to maintain a stable connection with the material bed at all times.

[0039] Specifically, to prevent short circuits caused by deformation and displacement, the steel wire used to make the metal spring 42 is coated with insulating varnish; the metal braided mesh is completely covered with an insulating sleeve.

[0040] Specifically, when the solid fermentation bed shrinks and its surface descends, the entire structure consisting of the probe rod 43, the lower positioning plate 23, the counterweight block 49, and the guide rod 22, under the combined action of its own weight and spring thrust (excluding spring thrust if metal braided strap 50 is used), moves vertically downwards with the bed surface, maintaining close contact between the tip of the probe rod 43 and the material, while simultaneously maintaining the continuity of the circuit through the metal spring 42 or the metal braided strap 50.

[0041] Among the optional solutions in this embodiment, the more preferred one is as follows: Figure 7 and Figure 8 As shown, each fixed external component 41 is fixed on the upper positioning plate 40. The top cover 2 has a through mounting hole, and the upper positioning plate 40 is fixedly inserted (note that it should be sealed) into the mounting hole.

[0042] Specifically, each fixed external component 41 can be directly fixed to the top cover 2, or it can be fixed to the upper positioning plate 40, and then fixed to the top cover 2 by the upper positioning plate 40 (the upper end of the fixed external component 41 is located outside the upper fermentation chamber, and the connecting joint 20 on it can be equipped with terminals and connected to the high-density resistivity detection system through the multi-core cable 19). The fixed external component 41 can be fixed by interference fit.

[0043] Specifically, here are the settings instructions for monitoring other parameters: Among the optional solutions in this embodiment, the more preferred one is as follows: Figures 9-14As shown, the top cover 2 is also equipped with multiple multi-point temperature measuring probes 33 and / or multiple imaging components (which can be reasonably selected and arranged according to actual needs); each multi-point temperature measuring probe 33 is distributed in a V-shape around the axis of the top cover 2, or each multi-point temperature measuring probe 33 is distributed in a straight line along the diameter direction passing through the center of the top cover 2; and the distance between the axis of each multi-point temperature measuring probe 33 and the axis of the top cover 2 is different; the lower end of each multi-point temperature measuring probe 33 is inserted into the fermentation material; each imaging component is distributed in a V-shape around the axis of the top cover 2, or each imaging component is distributed in a straight line along the diameter direction passing through the center of the top cover 2; and the distance between the axis of each imaging component and the axis of the top cover 2 is different; the imaging component includes a pointed blind tube 34 and an endoscope, the lower end of the pointed blind tube 34 is inserted into the fermentation material, the endoscope is placed inside the pointed blind tube 34, and the pointed blind tube 34 is transparent.

[0044] Specifically, for the multi-point temperature probe 33, since this probe needs to penetrate the material bed (fermented material, the same applies below, and will not be explained further) to carry out the detection, in order to avoid the dense arrangement causing serious impact on the physical properties of the material bed and the transfer of heat and moisture, a method can be adopted. Figure 10 and Figure 11 The arrangement shown is either a V-shape or a straight line to avoid the probe spacing being too small. The temperature signal of the multi-point temperature measuring probe 33 is introduced into one or more temperature monitoring instruments, and after being transmitted, it is introduced into a host computer for data recording.

[0045] Specifically, the same method can be used for the pointed blind tube 34. Figure 13 and Figure 14 The arrangement shown is to avoid the blind tubes being too close together. During fermentation, an endoscope can be inserted into the pointed blind tube 34, and the forward-facing lens or the rotatable side lens of the endoscope can be used to photograph the fine morphology of the material particles in the bed through the tube wall.

[0046] Specifically, the air distribution assembly is located at the bottom of the lower air chamber and includes a multi-layered annular air distribution pipe 7 to promote uniform distribution of airflow in the horizontal direction.

[0047] Specifically, the bottom of the lower chamber is equipped with a first temperature and humidity sensor 8, and its top is fitted with a waterproof cover to prevent contamination by liquid seeping from the fermentation material. Additionally, there is a first pressure measuring pipe 10 and a drain pipe 11, both equipped with valves that can be closed when not in use. A second temperature and humidity sensor 18 is arranged on the top cover 2. The exhaust pipe 17 has a valve, and a branch pipe on the exhaust pipe 17 serves as a second pressure measuring pipe 46. The connection method of the three differential pressure gauges arranged on the first pressure measuring pipe 10 and the second pressure measuring pipe 46 is as follows... Figure 5 As shown, the signals from the first temperature and humidity sensor 8 and the second temperature and humidity sensor 18, as well as the signals from each differential pressure gauge, are all introduced into the host computer for recording via signal lines.

[0048] Specifically, each support leg 13 of the reactor body 1 is equipped with a weight sensor 12 at its bottom. The signal is connected to the host computer for recording, which can obtain online data of the total weight of the entire reactor body 1 (including fermentation materials, etc.).

[0049] The following are the specifications regarding the multi-functional fermentation chamber 24: Specifically, the fermentation chamber has a two-compartment, three-section structure: the lower chamber and the middle chamber are integrated into one compartment. The lower chamber houses the solid-state bioreactor. An air purification unit consisting of a blower 27 and a sterilizing filter 25 is installed on the outer back of the middle chamber. A roller shutter 35 that can be opened at any angle is installed on the outer front of the middle chamber. When the top cover 2 of the solid-state bioreactor is open, a crane 29 (i.e., lifting equipment) can be used to lift or lower the top cover 2. The blower 27 of the air purification unit is turned on to form laminar flow, protecting the upper opening of the reactor body 1 from external pollutants. At the same time, there are two handholes 30 on each side of the middle chamber, with a fixed rubber glove installed on them, allowing manual addition, inoculation, stirring, or sampling of materials in the reactor body 1 using sterile tools, provided that clean air is turned on and the cover is open. The upper chamber is equipped with a crane 29 for lifting the top cover 2 and a heating component 28 for heating the inner cavity of the multifunctional fermentation chamber 24. The heating element 28 with a fan (which also includes a controller for better temperature control) can supply hot air to the middle and lower chambers to maintain a stable temperature within the chamber. The fermentation chamber is enclosed by panels 26 on all sides and top, except for a roller shutter 35 on the front of the middle chamber, an air purification unit on the back of the middle chamber (and the external connection port corresponding to the heating element 28), to ensure stable internal temperature. The multi-functional fermentation chamber 24 has casters 31 and retractable feet 32 ​​at its bottom for easy movement. The area around the casters 31 is not enclosed by panels 26, leaving a gap for excess airflow to escape.

[0050] The following three cases illustrate the implementation of the solid-state bioreactor based on Example 1: Case 1 The reactor body 1, made of polypropylene, is placed in the multifunctional fermentation chamber 24. A 3% hydrogen peroxide aqueous solution is sprayed onto the inner wall of the reactor body 1 facing the material, the surface of the sieve plate 3 and the top cover 2, as well as the inner wall of the air inlet pipe and the air distribution pipe 7, and maintained for 60 minutes. Afterwards, an ultraviolet lamp is placed inside the reactor body 1 for sterilization, and irradiation time is 60 minutes. No stainless steel wire is installed in the wire groove 5 on the sieve plate 3 inside the reactor body 1, and no lead wire 6 is connected. The number of high-density resistivity probes is 30.

[0051] The prepared solid fermentation substrate was pre-filled in a metal bucket, the opening of which was sealed with eight layers of pure cotton gauze. The bucket was then placed in a pressure sterilizer and sterilized at 121°C for 20 minutes. After cooling, the air purification unit of the multifunctional fermentation chamber 24 was turned on, the heating component 28 was turned off, and the roller shutter 35 in the front central cavity of the multifunctional fermentation chamber 24 was opened. The sterile substrate was then poured into the reactor body 1. Next, 100 mL of the cultured Beauveria bassiana liquid inoculum was poured onto the substrate. The roller shutter 35 was closed to 1 / 4 of its opening. Using the handhole 30 on the side of the multifunctional fermentation chamber 24 and the rubber glove fixed to the handhole 30, a pre-sterilized stirring spatula was held to thoroughly mix the solid substrate containing the inoculum and level the surface. After closing the top cover 2, insert the temperature probe (previously sterilized by soaking in a 3% hydrogen peroxide solution for 60 minutes) and the pointed blind tube 34 (previously sterilized at 121℃ for 20 minutes). Circulate water at a constant temperature of 30.0℃ into the jacket 14 through the water inlet 15. Simultaneously, sterile air at 28℃ and 95% relative humidity is introduced into the tank through the bottom air inlet at a flow rate of 25 L / min. After 72 hours of fermentation, data inversion is performed using a host computer, and the resulting resistivity contour plot is shown below. Figure 16 At 72 hours of fermentation, the temperature field distribution contour maps obtained by interpolation calculations of the exported temperature values ​​at various points are shown below. Figure 17 (In the figure, the vertical axis ρ represents the central axis of the cylindrical bed, and the horizontal axis r represents the radius). The temperature change trend at points 1-4 throughout the fermentation process is as follows: Figure 18 The four temperature measuring points on the temperature measuring probe closest to the center of the material bed are numbered 1, 2, 3, and 4 from bottom to top.

[0052] Case 2 The reactor body 1, made of polypropylene, was placed in the multifunctional fermentation chamber 24. A 0.5% peracetic acid aqueous solution was sprayed onto the inner wall of the reactor body 1 facing the material, the surface of the sieve plate 3 and the top cover 2, as well as the inner wall of the air inlet pipe and the air distribution pipe 7, and kept for 60 minutes. After that, an ultraviolet lamp was placed inside the reactor body 1 for disinfection, and the irradiation time was 60 minutes.

[0053] The prepared solid fermentation substrate was pre-filled in a metal bucket, the opening of which was sealed with eight layers of pure cotton gauze. The bucket was then placed in a pressure sterilizer and sterilized at 121°C for 20 minutes. After cooling, the air purification unit of the multi-functional fermentation chamber 24 was turned on, the heating component 28 was turned off, and the roller shutter 35 in the front central cavity of the multi-functional fermentation chamber 24 was opened. The sterile substrate was then poured into the reactor body 1. Next, 100 mL of the cultured Beauveria bassiana liquid inoculum was poured onto the substrate. The roller shutter 35 was closed to 1 / 4 of its opening. Using the handhole 30 on the side of the multi-functional fermentation chamber 24 and the rubber glove fixed to the handhole 30, a pre-sterilized stirring spatula was held to thoroughly mix the solid substrate containing the inoculum and level the surface. After closing the top cover 2, insert a temperature probe that has been disinfected by soaking in a 0.5% peracetic acid aqueous solution for 60 minutes and a pointed blind tube 34 that has been sterilized at 121℃ for 20 minutes. Circulate water at a constant temperature of 30℃ into the jacket 14 through the water inlet 15. Simultaneously, sterile air at 28℃ and 95% relative humidity is introduced into the tank through the bottom air inlet at a flow rate of 25 L / min. After 48 hours of fermentation, the interior of the material bed is photographed using an endoscope and the pointed blind tube 34 (which is transparent). Figure 19 (The red dots in the upper part of the diagram indicate the location of the endoscopic imaging point in the well logging; the rectangular plane in the diagram represents the semi-axial section of the material bed, the vertical axis is the central axis of the cylindrical material bed, the horizontal axis is the radius, and the origin is located at the center of the bottom surface of the material bed.) After fermentation, the cylindrical material bed is cut open, and the cross-sectional shape is as follows: Figure 20 .

[0054] Case 3 The reactor body 1, made of polypropylene, was placed in the multifunctional fermentation chamber 24. A 0.5% peracetic acid aqueous solution was sprayed onto the inner wall of the reactor body 1 facing the material, the surface of the sieve plate 3 and the top cover 2, as well as the inner wall of the air inlet pipe and the air distribution pipe 7, and kept for 60 minutes. After that, an ultraviolet lamp was placed inside the reactor body 1 for disinfection, and the irradiation time was 60 minutes.

[0055] The prepared solid fermentation substrate was pre-filled in a metal bucket, the opening of which was sealed with eight layers of pure cotton gauze. The bucket was then placed in a pressure sterilizer and sterilized at 121°C for 20 minutes. After cooling, the air purification unit of the multi-functional fermentation chamber 24 was turned on, the heating component 28 was turned off, and the roller shutter 35 in the front central cavity of the multi-functional fermentation chamber 24 was opened. The sterile substrate was then poured into the reactor body 1. Next, 100 mL of the cultured Beauveria bassiana liquid inoculum was poured onto the substrate. The roller shutter 35 was closed to 1 / 4 of its opening. Using the handhole 30 on the side of the multi-functional fermentation chamber 24 and the rubber glove fixed to the handhole 30, a pre-sterilized stirring spatula was held to thoroughly mix the solid substrate containing the inoculum and level the surface. After closing the top cover 2, insert a temperature probe that has been disinfected by soaking in a 0.5% peracetic acid aqueous solution for 60 minutes and a pointed blind tube 34 that has been sterilized at 121℃ for 20 minutes. Circulate water at a constant temperature of 30℃ into the jacket 14 through the water inlet 15. Simultaneously, sterile air at 28℃ is introduced into the tank through the bottom air inlet at a flow rate of 25 L / min, with the relative humidity fluctuating between 90% and 99%. The pressure difference across the bed and the total weight of the reactor body 1 and its contents over the entire 120-hour fermentation process are shown in the following figures. Figure 21 .

[0056] Example 2 This embodiment provides a method for using the solid-state bioreactor based on Embodiment 1, including the following steps: S1. Place the solid-state bioreactor in the lower chamber of the multifunctional fermentation chamber 24 and disinfect the interior of the solid-state bioreactor. (When the reactor body 1 and top cover 2 are made of metal or metal-lined polymer coating, the interior of the reactor body 1 can be disinfected directly with steam. Steam is introduced from the bottom gas distribution pipe 7 and discharged from the exhaust pipe 17 on the top cover 2. If the reactor body 1 or top cover 2 is made of polymer material, the interior of the reactor body 1 should be chemically disinfected. Hydrogen peroxide, ethanol or peracetic acid aqueous solution can be sprayed on the inner wall of the reactor body 1 facing the material, the surface of the sieve plate 3 and the top cover 2, as well as the inner wall of the air inlet pipe and the gas distribution pipe 7, and kept for at least 30 minutes. Alternatively, an ultraviolet lamp can be placed inside the reactor body 1 for disinfection for at least 30 minutes.) S2, turn on the air purification unit, turn off the heating component 28, open the roller shutter 35 and put the sterilized and cooled solid fermentation substrate (prepared solid fermentation substrate in a metal bucket, the bucket opening is sealed with eight layers of pure cotton gauze, placed in a pressure sterilizer, sterilized at 121℃ for 20 min and then cooled, the roller shutter 35 is opened and poured into the reactor body 1) onto the sieve plate 3, pour in the inoculum (pour a small amount of cultured liquid inoculum or solid inoculum suspended in sterile water onto the solid fermentation substrate), reduce the opening of the roller shutter 35, and use a sterile tool (pre-sterilized stirring tool (stick or shovel)) held with rubber gloves to stir the solid fermentation substrate mixed with inoculum evenly and spread the surface evenly. S3, close the top cover 2 (the multi-point temperature measuring probe 33 and the pointed blind tube 34 can be inserted after the top cover 2 is closed, and they also need to be disinfected or sterilized beforehand), turn on the temperature control component (which is a jacket 14, and it is equipped with a water inlet 15 and a water outlet 16; constant temperature circulating water is introduced into the jacket 14 through the water inlet 15), and sterile air regulated by temperature and humidity is introduced into the lower air chamber through the air distribution component according to the predetermined flow rate; S4, turn off the air purification unit and the roller shutter 35, start the heating component 28 to control the temperature of the inner cavity of the multi-functional fermentation chamber 24, and start fermentation; and use the host computer connected to the high-density resistivity detection system to monitor (monitor the equipment according to the various parameters set, and use the host computer to record the tank weight (i.e., the data detected by the weight sensor 12), the temperature and humidity of the inlet and outlet air, the pressure and pressure difference, and the multi-channel temperature data measured by the temperature probe inside the material bed, etc.; and the detection of high-density resistivity imaging can be carried out at any time during the fermentation process, and the data inversion and recording functions are all realized through the host computer).

[0057] Through the coordinated operation of the solid-state bioreactor and the 24 components of the multifunctional fermentation chamber, aseptic and precise control and real-time monitoring of the fermentation status are achieved throughout the entire solid-state fermentation process. The solid-state bioreactor is disinfected in the early stages, and the air purification unit is activated during the inoculation and loading phases, with the use of rubber gloves to create a reliable localized aseptic environment, significantly reducing the risk of contamination. During the fermentation phase, the temperature control and heating components 28 are activated simultaneously, combined with the gas distribution components to introduce temperature- and humidity-controlled aseptic air, achieving precise control of the fermentation temperature and ventilation environment. During fermentation, a high-density resistivity detection system linked to a host computer monitors the fermentation status in real time, effectively preventing localized overheating and water loss in the material bed, and ensuring stable fermentation quality.

[0058] Specific examples have been used to illustrate the principles and implementation methods of this invention. The descriptions of the above embodiments are only for the purpose of helping to understand the method and core ideas of this invention. Furthermore, those skilled in the art will recognize that, based on the ideas of this invention, there will be changes in the specific implementation methods and application scope. Therefore, the content of this specification should not be construed as a limitation of this invention.

Claims

1. A solid-state bioreactor, characterized in that: include: A solid-state bioreactor includes a reactor body. The inner cavity of the reactor body is divided into an upper fermentation chamber and a lower gas chamber by a sieve plate. The upper fermentation chamber is located above the lower gas chamber. The sieve plate is used to support the fermentation material. A temperature control component corresponding to the position of the upper fermentation chamber is provided on the outer wall of the reactor body. An air distribution component for introducing sterile air is provided in the lower gas chamber. A top cover is closed at the upper opening of the reactor body. An exhaust pipe and at least four high-density resistivity probes are fixed on the top cover. A drain pipe is provided at the bottom of the lower gas chamber. The top cover is coaxially arranged with the reactor body. The high-density resistivity probes are arranged at equal intervals along the diameter direction passing through the center of the top cover. The lower part of each high-density resistivity probe is used to insert into the surface of the fermentation material at an equal depth. Each high-density resistivity probe is connected to a high-density resistivity detection system. An insulating layer is provided on the surfaces of the upper fermentation chamber, the top cover, and the sieve plate facing the fermentation material. The multifunctional fermentation chamber has an internal cavity divided into an upper cavity, a middle cavity, and a lower cavity that are interconnected. The lower cavity is used to house the solid bioreactor, and the bottom of the lower cavity has a lower opening. The middle cavity has multiple hand holes on its side wall, and rubber gloves are provided for the hand holes. The middle cavity side wall is also equipped with an adjustable roller shutter that is connected to the outside. An air purification unit is provided on the middle cavity side wall opposite to the roller shutter. In the vertical direction, the upper opening of the reactor body is located in the horizontal airflow area between the air purification unit and the roller shutter. The upper cavity is equipped with a lifting device and a heating component. The lifting device is used to lift the top cover, and the upper cavity has an external connection port that connects to the outside.

2. The solid-state bioreactor according to claim 1, characterized in that: The high-density resistivity probe includes a fixed external component, a probe telescopic cylinder, a probe rod, and an electrical connector. The fixed external connector can be fixedly connected to the top cover, and the upper end of the fixed external connector is connected to the high-density resistivity detection system through a wire; The probe telescopic cylinder can be fixed to the top cover inside the upper fermentation chamber, and the lower end of the fixing external component is located inside the inner cavity of the probe telescopic cylinder; the probe telescopic cylinder is made of non-conductive material; The upper end of the probe rod is slidably disposed in the inner cavity of the probe telescopic cylinder in a vertical direction, and the lower end of the probe rod is used to insert into the surface layer of the fermentation material. The energized connector is located inside the probe telescopic cylinder between the fixed external connector and the probe rod, and the energized connector is used to energize the fixed external connector and the probe rod.

3. The solid-state bioreactor according to claim 2, characterized in that: The lower end of each of the probe rods is fixed to the lower positioning plate; the lower positioning plate is located inside the upper fermentation chamber, and in the vertical direction, the lower positioning plate is higher than the lower end of the probe rod.

4. The solid-state bioreactor according to claim 3, characterized in that: The top cover is provided with multiple guide components, each guide component having a guide rod that extends and retracts in the vertical direction, and the lower positioning plate is fixedly connected to the lower end of each guide rod.

5. The solid-state bioreactor according to claim 3, characterized in that: The electrically connected component is a metal spring, and the metal spring is in a compressed state located between the fixed external component and the probe rod.

6. The solid-state bioreactor according to claim 3, characterized in that: The power-conducting connector is a metal braided strip, and the lower positioning plate is reinforced.

7. The solid-state bioreactor according to claim 2, characterized in that: Each of the aforementioned fixed external components is fixed to the upper positioning plate, and the top cover has a through mounting hole, and the upper positioning plate is fixedly inserted into the mounting hole.

8. The solid-state bioreactor according to claim 1, characterized in that: The top cover is also equipped with multiple multi-point temperature measuring probes and / or multiple imaging components; The multi-point temperature measuring probes are distributed in a V-shape around the axis of the top cover, or in a straight line along the diameter direction passing through the center of the top cover; and the distance between the axis of each multi-point temperature measuring probe and the axis of the top cover is different; the lower end of each multi-point temperature measuring probe is inserted into the fermentation material. Each of the imaging components is distributed in a V-shape around the axis of the top cover, or each of the imaging components is distributed in a straight line along the diameter direction passing through the center of the top cover; and the distance between the axis of each imaging component and the axis of the top cover is different; each imaging component includes a pointed blind tube and an endoscope, the lower end of the pointed blind tube is inserted into the fermentation material, and the endoscope is placed inside the pointed blind tube.

9. The solid-state bioreactor according to claim 1, characterized in that: The sieve plate has a groove in the center, and stainless steel wires are arranged in the groove. The side wall of the lower air chamber is provided with an insulating terminal. One end of the stainless steel wire is connected to the insulating terminal through a lead wire. The insulating terminal is connected to the dedicated terminal of the high-density resistivity detection system through a wire.

10. A method of using a solid-state bioreactor based on any one of claims 1 to 9, characterized in that: Includes the following steps: S1, The solid bioreactor is placed in the lower cavity of the multifunctional fermentation chamber, and the interior of the solid bioreactor is disinfected; S2, turn on the air purification unit, turn off the heating component, open the roller shutter and load the sterilized and cooled solid fermentation substrate onto the sieve plate, pour in the inoculum, reduce the opening of the roller shutter, and use the rubber gloves to hold the sterile tool to stir the solid fermentation substrate mixed with the inoculum evenly. S3, close the top cover, turn on the temperature control component, and introduce sterile air regulated by temperature and humidity into the lower air chamber through the air distribution component at a predetermined flow rate; S4, turn off the air purification unit and the roller shutter, start the heating component to control the temperature of the inner cavity of the multifunctional fermentation chamber, and start fermentation; and use the host computer that is communicatively connected to the high-density resistivity detection system to monitor it.

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