Compound enzyme preparation device and method and application of compound enzyme preparation device and method in efficient removal of dental plaque

Abstract

The invention relates to the technical field of compound enzymes, in particular to a compound enzyme preparation device and method and application of the compound enzyme preparation device and method to efficient dental plaque removal, and solves the defects in the prior art. The compound enzyme preparation device comprises a base table, a fermentation tank, an adjusting structure, a drainage structure and a culture structure; a connecting table is arranged on the supporting frames on the two sides, an extending table and an L-shaped support are further arranged outside the supporting frames on the two sides respectively, and the fermentation tank is fixed to the middle of the top face of the connecting table. Compared with the prior art, the preparation efficiency of compound enzyme can be effectively improved, the automation level of material supplementing in the process can be improved, and the production efficiency is improved. And defects possibly caused by human factors are avoided.

Description

Technical Field

[0001] This invention relates to the field of complex enzyme technology, and in particular to a complex enzyme preparation apparatus, method, and its application in the efficient removal of dental plaque. Background Technology

[0002] Enzymatic plaque removal is an emerging biological strategy that enhances removal effectiveness by specifically degrading plaque matrix components (such as dextran) and disrupting the integrity of the biofilm. Dextranase is an enzyme that catalyzes the hydrolysis of α-1,6 glycosidic bonds in dextran, effectively breaking down the glucan network and weakening the adhesion and stability of plaque. However, the catalytic efficiency of single dextranases is limited and they are easily affected by the oral environment (such as pH, temperature, and inhibitors), leading to unsatisfactory cleaning results. Furthermore, natural dextranases may suffer from low enzyme activity, poor stability, and high cost, limiting their large-scale application.

[0003] Furthermore, the market currently lacks efficient preparation equipment and methods specifically for compound enzymes (especially those based on dextranase), resulting in unstable performance and high production costs for compound enzyme preparations, which hinders their widespread application in oral care products (such as toothpaste and mouthwash).

[0004] Therefore, we propose a compound enzyme preparation device and method, and its application in the efficient removal of dental plaque, to solve the problems mentioned above. Summary of the Invention

[0005] The purpose of this invention is to provide a compound enzyme preparation device and method, and its application in the efficient removal of dental plaque, so as to solve the problems mentioned in the background art.

[0006] To solve the above-mentioned technical problems, the present invention provides the following technical solution: A compound enzyme preparation device includes a base, a fermenter, an adjustment structure, a flow guiding structure, and a culture structure. The base is provided with two sets of support frames, and a connecting platform is provided on the two side support frames. An extension platform and an L-shaped bracket are also provided on the outside of the two side support frames respectively. The fermenter is fixed in the middle position on the top surface of the connecting platform. The adjustment structure is provided in two sets, both of which are rotatably mounted between the side walls of the two support frames. The adjustment structure holds a triangular bottle that can swing and adjust accordingly. The drainage structure is slidably mounted on the extension platform, and both sides of the drainage structure are equipped with drainage tubes that can be raised and lowered for adjustment. The culture structure is mounted on an L-shaped support. A growth culture tube connects the culture structure to the fermenter. A valve for controlling flow regulation is installed in the middle section of the growth culture tube. The fermenter is equipped with a fixed tube and a piston tube that are adjusted according to the dissolved oxygen level inside the tank. The piston tube slides along the inner wall of the fixed tube. A transmission component for controlling the rotation of the valve is also installed between the piston tube and the valve.

[0007] Compared with the prior art, the beneficial effects achieved by the present invention are: 1. By setting up fermenters, lifting components, adjustment structures, culture structures, and drainage structures, the compound enzyme preparation process can be realized in a semi-automatic manner, which can effectively improve the preparation efficiency and avoid the drawbacks that may be caused by human factors.

[0008] 2. By setting up a fixed tube, piston tube, transmission components, and valve in the culture structure, the change in air pressure inside the fermenter is used as the power to regulate the movement of the piston tube. When the dissolved oxygen content inside the tank rises, its internal air pressure is higher than that outside, causing the piston tube to make linear displacement movement inside the fixed tube. Then, through the cooperation of the transmission components, the valve opens accordingly and controls the flow rate of the growth medium. When the valve opens to its maximum angle, it will not be affected by the piston tube continuing to move outward until the concentration sensor is triggered and the feeding process of the growth medium stops, effectively realizing the automated feeding of the growth medium. Attached Figure Description

[0009] The technical solution and other beneficial effects of this application will become apparent from the following detailed description of specific embodiments in conjunction with the accompanying drawings.

[0010] In the attached diagram: Figure 1 This is a schematic diagram of the overall structure of the present invention; Figure 2 yes Figure 1 A cross-sectional view along the central axis of the fermenter; Figure 3 This is a schematic diagram of the installation of the adjustment structure of the present invention; Figure 4 This is a schematic diagram of the adjusting structure of the present invention; Figure 5 This is a schematic diagram of the installation of the culture structure of the present invention; Figure 6 yes Figure 5 A top-view structural diagram; Figure 7 This is a schematic diagram of the structure of the fixed tube and piston tube of the present invention; Figure 8 This is a schematic diagram of the structure of the component on the guide rail platform of the present invention; Figure 9 This is a schematic diagram of the drainage structure of the present invention; Figure 10 This refers to the content of each component in YNB as described in the embodiments of this invention.

[0011] In the diagram: 1. Base; 11. Control panel; 2. Support frame; 21. Connecting platform; 22. Extension platform; 221. Screw 1; 23. L-shaped bracket; 24. Transmission gear; 3. Fermentation tank; 31. Inoculation tube; 311. Adjusting tube; 32. Ventilation tube; 33. Growth culture tube; 34. Induction culture tube; 4. Lifting assembly; 41. Lifting motor; 42. Connecting rod; 43. Connecting plate; 5. Adjustment structure; 51. Fixing frame; 52. Adjusting frame; 53. Screw 2; 54. Fixing plate; 55. Rotating frame; 551. U-shaped frame; 552. Positioning clamp; 553. Heating element; 554. 56. Limiting shaft; 57. Rotating handle; 58. Adjusting handle; 59. Connecting block one; 50. Rotating shaft; 60. Culture structure; 61. T-tube; 62. Guide rail platform; 621. Gear condition one; 622. Rack block; 623. Piston rod; 624. Limiting seat; 625. Positioning block; 626. Connecting frame; 627. Gear condition two; 63. Fixed tube; 631. Keyway; 64. Piston tube; 641. Key bar; 65. Adjusting gear one; 66. Adjusting gear two; 661. Valve disc; 70. Drainage structure; 71. Sliding seat; 72. Lifting platform; 73. Pump body two; 731. Drainage tube. Detailed Implementation

[0012] The following disclosure provides many different embodiments or examples for implementing different structures of this application. To simplify the disclosure, specific examples of components and arrangements are described below. Of course, these are merely examples and are not intended to limit the scope of this application. Furthermore, reference numerals and / or letters may be repeated in different examples; such repetition is for simplification and clarity and does not in itself indicate a relationship between the various embodiments and / or arrangements discussed. In addition, various specific examples of processes and materials are provided in this application, but those skilled in the art will recognize the application of other processes and / or the use of other materials.

[0013] like Figure 1As shown, the present invention provides a technical solution: a compound enzyme preparation device, including a base, an adjustment structure 5, a culture structure 6, and a drainage structure 7. The base includes a base 1, a support frame 2, an extension platform 22, and an L-shaped support 23. A control panel 11 with a PLC controller is fixedly installed on the base 1. Two support frames 2 are symmetrically arranged and are both fixed to the base 1 by bolts. Two open and symmetrical fixing holes are opened on one of the support frames 2 (the fixing holes are arranged along the length direction of the support frame 2). A connecting platform 21 is also fixed between the top walls of the two support frames 2 by bolts. The bottom end of the L-shaped support 23 is fixed to the base 1 by bolts and is located on one side of one of the support frames 2. The extension platform 22 is also fixed to the base 1 by bolts and is also connected and fixed together with the support frame 2 on the other side.

[0014] like Figure 2-3 As shown, two inner connecting plates 1 at the same height are fixed between the opposite side walls of the two support frames 2 by bolts. One inner connecting plate 2 is also provided below the inner connecting plate 1 and is also fixed between the side walls of the two support frames 2. A transmission gear 24 is rotatably connected between the top surface of the inner connecting plate 1 and the bottom surface of the connecting platform 21 by a shaft. A motor 1 (motor 1 is a rotary motor) is fixedly installed on the bottom wall of the inner connecting plate 2. The output shaft of the motor 1 drives the transmission gear 24 located above it, and the transmission gears 24 on both sides mesh with each other.

[0015] It should be further explained that by starting motor one, it is used as a drive source to control the transmission gear 24 connected to it to rotate. The other transmission gear 24 is meshed with the first transmission gear 24, so that the other transmission gear 24 rotates synchronously. The two transmission gears 24 rotate in opposite directions, and the rotation angle of the transmission gear 24 is 90 degrees.

[0016] like Figure 3-4 As shown, the adjustment structure 5 includes a fixed frame 51, an adjustment frame 52, a rotating frame 55, and an output component; The fixing frame 51 is installed below the bottom wall of the inner plate and is coaxially connected to the transmission gear 24 located above it. The fixing frame 51 has a groove, and guide grooves (dovetail type) are provided on both sides of the groove. Two identical screws 53 are rotatably installed in the groove (a motor that drives the screws 53 is installed between the inner walls of the fixing frame 51, not shown in the figure). An adjusting frame 52 is slidably installed along the inner wall of the groove. Guide bars (dovetail type) are fixedly installed on both sides of the adjusting frame 52. The guide bars slide between the inner walls of the guide grooves and never disengage. The end of the screw 53 is always located inside the adjusting frame 52 and the screw 53 is connected to the inner wall of the adjusting frame 52 by threaded transmission.

[0017] A second groove is provided on the adjusting frame 52, and a second motor (a rotary motor) is fixedly installed in the second groove. Fixing plates 54 are bolted to both sides of the top surface of the adjusting frame 52 above the second groove. The fixing plates 54 have arc-shaped holes. A rotating shaft 581 is rotatably mounted between the side walls of the two fixing plates 54 via bearings. Two connecting blocks 58 are fixedly connected to the rotating shaft 581, and the two connecting blocks 58 are fixed to the bottom wall of the rotating frame 55. A limiting shaft 554 passes through the inner wall of the rotating frame 55, sliding between the arc-shaped holes on the two fixing plates 54. The rotating frame 55 has a "U"-shaped structure, and a heating element 553 is fixedly installed on the concave surface of the rotating frame 55. 3 is a plate structure with a built-in heating rod (the outer shell of the plate is made of heat-conducting metal). A switch for controlling the heating element 553 is also installed on the outer arc surface of the rotating frame 55. A U-shaped frame 551 is rotatably connected to the end of the rotating frame 55 via a shaft (a motor 3, not shown in the figure, is installed inside the end of the rotating frame 55 to drive the rotation of the U-shaped frame 551). A "T"-shaped piston shaft is elastically provided in the middle of the U-shaped frame 551 (the spring on the piston shaft is located between the positioning clamp 552 and the opposite side wall of the U-shaped frame 551). One end of the piston shaft faces the direction of the arc surface of the rotating frame 55, and the other end of the piston shaft is fixedly connected to the arc-shaped positioning clamp 552. A sponge pad is adhered to the surface of the positioning clamp 552 that contacts the triangular bottle.

[0018] The output component includes a rotating handle 56, an adjusting handle 57, and a rotating shaft 581. The adjusting handle 57 has a fixing hole 2 along its length direction, and one end of the adjusting handle 57 is connected to the rotating shaft 581 as a whole. The rotating handle 56 is fixedly connected to the end of the output shaft of the second motor. A guide bolt is also threadedly connected to the side wall of the rotating handle 56 facing the adjusting handle 57. The guide bolt slides between the inner walls of the fixing hole 2.

[0019] It should be further explained that the movement of the two meshing transmission gears 24 synchronously drives the two fixed brackets 51 below to rotate and adjust. When the transmission gears 24 rotate at an angle of 90 degrees, the two fixed brackets 51 just rotate into the fixing hole and contact one of the inner walls. In this state, the activated bacterial solution in one triangular bottle is transferred to the other triangular bottle for further cultivation through the drainage structure 7.

[0020] During the cultivation process, the Erlenmeyer flask is placed in the middle of the heating element 553. Then, by starting motor three, the U-shaped frame 551 is rotated to the side where the Erlenmeyer flask is located until the positioning clamp 552 contacts and clamps the wall of the Erlenmeyer flask. Then, motor three is stopped. When shaking the flask, motor two is started, which drives the rotating handle 56 to rotate. This causes the guide bolt on the rotating handle 56 to slide back and forth between the inner walls of the fixing hole two, thereby causing the adjusting handle 57 to swing back and forth 60 degrees. This, in turn, drives the rotating frame 55 to swing back and forth synchronously through the rotating shaft 581, so that the bacterial solution in the Erlenmeyer flask is shaken and mixed evenly under heating. After the process is completed, motor three is stopped.

[0021] like Figure 1-2 as well as Figure 5 As shown, a fermenter 3 is bolted to the middle position on the connecting platform 21. The fermenter 3 has a lid on top, and a concentration sensor is embedded in the inner wall of the fermenter 3. A stirrer (not shown in the figure) is also installed on the inner bottom wall. Four electric cylinders are installed on the top surface of the connecting platform 21. A U-shaped rod is connected to the output shaft end of the electric cylinder. The other end of the U-shaped rod is connected and fixed to the top surface of the lid. An "L"-shaped inoculation tube 31 is installed on one side wall of the fermenter 3. A vertically set support column is bolted to the top surface of the connecting platform 21. A pump body is fixedly connected to the top of the support column. An adjusting tube 311 is threaded to the end of the inoculation tube 31 located outside the fermenter 3. A rigid tube (the rest is a flexible tube) is connected to the end of the adjusting tube 31 away from the inoculation tube 31. The inlet and outlet of the pump body are both connected to the flexible part of the adjusting tube 311.

[0022] Three connection ports are provided on the other side wall of the fermenter 3. A vent pipe 32, a growth culture tube 33, and an induction culture tube 34 are respectively threaded to the three connection ports. An electric one-way valve (not shown in the figure) is installed on the port of the connection port facing the fermenter 3. The electric one-way valve is fixed on the inner wall of the fermenter 3.

[0023] A lifting assembly 4 is also provided on the base 1. The lifting assembly 4 includes a lifting motor 41, a connecting rod 42 and a connecting plate 43. The lifting motor 41 is fixed to the base 1 by bolts. One end of the connecting rod 42 is fixed to the output shaft end of the lifting motor 41, and the other end of the connecting rod 42 is threadedly connected to the connecting plate 43. The top end of the rigid tube of the regulating tank is stuck between the inner wall of the connecting plate 43 and connected and fixed thereto.

[0024] It should be further explained that during the process of introducing the secondary cultured bacterial solution into fermenter 3, adjustments were made... Figure 1The position of the adjustment structure 5 on the left side is moved to the initial position (that is, the position corresponding to the rotation angle of the transmission gear 24 being 0 degrees). At this time, the mouth of the triangular bottle on the adjustment mechanism is exactly below the rigid tube of the adjustment tube 311. Then, by starting the lifting motor 41, it drives the rigid tube of the adjustment tube 311 downward into the triangular bottle through the connecting plate 43. Then, the pump body 1 is started, so that the bacterial liquid in the triangular bottle is introduced into the fermenter 3. After the end, the pump body 1 is turned off, and the connecting plate 43 is reset upward, so that the rigid tube of the adjustment tube 311 is moved out of the triangular bottle.

[0025] like Figure 5-8 As shown, the culture structure 6 includes a T-shaped tube 61, a guide rail 62, a fixed tube 63, a piston tube 64, a transmission assembly, and a valve disc 661. The transmission assembly includes an adjusting gear 1 65, an adjusting gear 2 66, a gear condition 1 621, and a gear condition 2 627. The bottom end of the T-shaped tube 61 is fixed to the top surface of the L-support 23. The growth culture tube 33 is screwed onto the end of the T-shaped tube 61. A valve disc 661 is rotatably arranged between the inner walls of the T-shaped tube 61, and an adjusting gear 66 is rotatably arranged on the top of the T-shaped tube 61. The adjusting gear 66 and the valve disc 661 are coaxially connected.

[0026] A connecting seat is fixedly installed on the side wall of the fermenter 3. One end of the fixing tube 63 passes through the connecting seat and extends to the inner wall of the fermenter 3 by means of a threaded connection. At least two keyways 631 are opened on the inner wall of the fixing tube 63. At least two key bars 641 are fixedly connected to the outer surface of one end of the piston tube 64. The key bars 641 slide between the inner walls of the keyways 631. One end of the piston tube 64 slides between the inner walls of the fixing tube 63. A connecting bolt 1 is fixedly connected to the outer surface of the fixing tube 63. A connecting bolt 2 is fixedly connected to the outer surface of the piston tube 64. A fixing hole 3 matching the connecting bolt 2 is also opened on the fixing tube 63. The fixing hole 3 is set to open towards one end of the piston tube 64. A tension spring is connected between the connecting bolt 1 and the connecting bolt 2. Furthermore, several toothed grooves are also opened along the length direction on the outer surface of the piston tube 64.

[0027] It should be further explained that gas is introduced into the fermenter 3 through the vent pipe 32 to keep the dissolved oxygen in the tank above 25% (25% is used as an example in this embodiment). As the reaction in the fermenter 3 proceeds, the dissolved oxygen continues to rise, which leads to an increase in the gas pressure in the fermenter 3. This, in turn, pushes the piston pipe 64 outward, thereby causing the adjusting gear 65 to rotate synchronously.

[0028] The guide rail platform 62 is fixed to the top surface of the L-bracket 23 by bolts. A groove three is provided on the guide rail platform 62 along its length. A fixed shaft is fixed between the inner walls of the groove three. A guide bar two is slidably arranged between the inner walls of the groove three. The fixed shaft passes through the guide bar two and slides in contact with it. A tooth condition one 621 is provided on the top surface of the guide rail platform 62. The guide bar two is fixed on the bottom wall of the tooth condition one 621 and is set to the same length as it. An adjusting gear one 65 is also provided between the tooth condition one 621 and the fixed tube 63. The adjusting gear one 65 is rotatably set on the top surface of the L-bracket 23 through a shaft connection. The adjusting gear one 65 is also meshed between the tooth condition one 621 and the tooth groove of the fixed tube 63. A tooth condition two 627 is also installed on the top surface of the tooth condition one 621 through two "L"-shaped connecting brackets 626. The tooth condition two 627 is meshed with the adjusting gear two 66.

[0029] It should be further explained that as the adjusting gear 65 rotates, the gear 621 meshing with it on the other side moves accordingly, thereby synchronously driving the gear 627 to move and adjust. As the gear 627 moves, the adjusting gear 66 rotates accordingly, and then drives the valve 661 in the T-tube 61 to rotate and adjust through the coaxial connection. This allows the pipeline connected to the growth culture tube 33 to flow the growth culture medium into the fermenter 3. The flow rate of the culture medium is controlled by the rotation angle of the valve 661. When the bacterial solution in the fermenter 3 reaches the set concentration, the feed pump of the growth culture medium is turned off. After the Pichia pastoris has completely consumed the glycerol in the culture medium and the substrate is kept in a state of scarcity for about 1 hour, the induction temperature is set, the induction culture tube 34 is opened, and the induction culture medium is fed in to induce dextranase expression.

[0030] A limiting seat 624 is fixed to the top surface of the guide rail 62 by screws. A positioning block 625 is fixed to the side wall of the tooth condition 621 by bolts. A positioning groove is provided on the side wall of the limiting seat 624 facing the positioning block 625 (a silicon steel sheet is fixed on the inner wall of the positioning groove). The positioning groove matches the positioning block 625 and they can be connected and fixed by suction. An external power supply is installed on one side of the limiting seat 624 on the side wall of the guide rail 62. The external power supply energizes the silicon steel sheet in the positioning groove, thereby generating suction on the silicon steel sheet so as to firmly attract the iron positioning block 625.

[0031] A rack block 622 is movably mounted on one side of the gear condition 621 via a shaft connection. A connecting block 2 is fixedly connected to the side wall of the gear condition 621, and a connecting block 3 is fixedly connected to the end wall of the rack block 622. A piston rod 623 (in the shape of an "I") passes through the connecting block 2 and the connecting block 3, and a spring is fitted on the piston rod 623. The two ends of the spring are respectively connected and fixed to the side walls of the connecting block 2 and the connecting block 3. When the rack block 622 contacts the gear condition 621, the distance between the connecting block 2 and the connecting block 3 is the closest, and the rack block 622 and the gear condition 621 form a tooth groove. When the distance between the connecting block 2 and the connecting block 3 is the farthest, the distance between the rack block 622 and the gear condition 621 is just enough to allow two adjacent teeth on the adjusting gear 65 to rotate (that is, this distance forms two adjacent tooth grooves).

[0032] It should be further explained that as the valve disc 661 opens, after rotating 90 degrees, the valve disc 661 is in a position parallel to the length direction of the growth culture tube 33. At this time, the flow rate is at its maximum, and the adjusting gear 65 is also rotating to mesh with the rack block 622. The positioning block 625 also moves into the limiting seat 624 and is connected to it by suction. That is, the gear condition 621 no longer moves, and the transmission gear 24 no longer rotates. When the dissolved oxygen in the fermenter 3 continues to rise, it will still drive the adjusting gear 65 to rotate. As the adjusting gear 65 continues to rotate, it continuously meshes with the rack block 622 and no longer exerts force on the gear condition 621 until the dissolved oxygen in the fermenter 3 stabilizes. If the stabilized dissolved oxygen decreases and recovers, under the action of the tension spring, the piston tube 64 moves into the fixed tube 63 to reset, thereby indirectly driving the valve disc 661 to close.

[0033] like Figure 9 As shown, the drainage structure 7 is slidably mounted on the extension platform 22. The drainage structure 7 includes a sliding seat 71, a lifting platform 72, a second pump body 73, and a drainage pipe 731. A groove 4 is provided on the extension platform 22 along its length. A screw 221 is rotatably mounted in the groove 4 by a motor 4. The motor 4 is built into the inner wall of the extension platform 22 (the motor 4 is a rotary motor). The sliding seat 71 has a "T" shape and slides between the inner walls of the groove 4. The screw 221 passes through the sliding seat 71 and is threadedly driven to it. A lifting cylinder is fixedly mounted on the top surface of the sliding seat 71. The lifting platform 72 is fixedly mounted on the output shaft end of the lifting cylinder. A second pump body 73 is also fixedly mounted on the lifting platform 72. Two "L"-shaped drainage pipes 731 are connected to the inlet and outlet of the second pump body 73, respectively.

[0034] It should be further explained that the adjustment structures 5 on both sides are rotated to a position close to the extension platform 22 by the transmission gear 24 (i.e., the adjustment structure 5 rotates 90 degrees), and the position of the adjustment frame 52 is adjusted so that the mouth of the triangular bottle is moved directly below the drainage tube 731. Then, the height of the lifting platform 72 is adjusted by activating the lifting cylinder so that the drainage tubes 731 on both sides extend into the interior of the triangular bottles on both sides. Then, the activated bacterial solution in one triangular bottle is transferred to the other triangular bottle by activating the pump body 73 for subsequent cultivation. After the transfer is completed, the adjustment structure 5 is rotated 90 degrees to the initial position and stopped to start the subsequent cultivation process.

[0035] Working principle: Activation culture: Prepare 30 mL of YPD medium [YPD activation medium (g / L): glucose 20, peptone 20, yeast extract 10, pH 5.5] and put it into a 250 mL Erlenmeyer flask. Autoclave at 115℃ for 15 min. After the medium cools to room temperature, pick 1 loopful of cells from the recombinant Pichia pastoris slant medium in a clean bench and inoculate it into the YPD medium. Fix it on the rotating rack 55 of one of the adjustment structures 5. Control the temperature of the heating element 553 to 30℃ and set the speed of motor 2 to 200 r / min. Activate the medium for 24 hours.

[0036] Cell culture: Prepare 30 mL of BMGY medium [BMGY growth medium (g / L): peptone 20, yeast extract 10, YNB 13.4 (composition see appendix)]. Figure 10 10g of glycerol, 0.0004g of biotin, and 0.1 mol / L potassium phosphate buffer (pH 6.0) were placed in another 250 mL Erlenmeyer flask and autoclaved at 115°C for 15 min. After the culture medium cooled to room temperature, the Erlenmeyer flask was fixed on the rotating rack 55 of another regulating structure 5. The 10% (v / v) activated bacterial solution was transferred to the other Erlenmeyer flask (i.e., BMGY medium) through the drainage structure 7. The temperature of the heating element 553 was controlled at 30°C, and the speed of the second motor was set to 200 r / min. The culture was continued for 24 hours.

[0037] High-density culture: Prepare 3L of BSM medium [BSM medium (g / L): CaSO4 0.93, K2SO4 18.2, MgSO4·7H2O 14.9, KOH 4.13, glycerol 40, 85% phosphate 26.7 mL / L, PTM1 4.35 mL / L, pH adjusted to 5.5 with 25% ammonia] in a 5L fermenter 3. Calibrate the pH electrode and dissolved oxygen electrode using calibration solution and assemble them. Finally, wrap the electrode protective sleeve with kraft paper and sterilize at 115℃ for 20min (standard operating procedure, so not shown in the figure). After the medium cools to room temperature, inoculate the BMGY bacterial solution into the 3L BSM medium at a 10% inoculation rate through inoculation tube 31. Adjust the aeration rate through aeration tube 32 to maintain dissolved oxygen above 25%. Set the temperature to 30℃ and the rotation speed to 500 r / min. When dissolved oxygen rises rapidly, the feed culture medium [growth medium: 50% (w / v) glycerol (containing 12 mL / L PTM1)] is started to be added to fermenter 3 by rotating valve 661. When the concentration sensor detects that the cell concentration has reached the set concentration, the feed culture pump is turned off. After the Pichia pastoris has completely consumed the glycerol in the culture medium and the substrate is kept in a state of depletion for 1 hour, the induction temperature is set and the induction culture tube 34 is started to feed the induction medium [induction medium: 100% methanol (containing 12 mL / L PTM1)] to induce dextranase expression.

[0038] After the composite enzyme is prepared by the above-described apparatus and method, it can be used to remove biofilms in the oral cavity.

[0039] In the description of this application, it should be noted that, unless otherwise expressly specified and limited, the terms "installation," "connection," and "linking" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection, an electrical connection, or a connection that allows communication between them; they can refer to a direct connection, the internal communication between two components, or the interaction between two components. Those skilled in the art can understand the meaning of the above terms in this application according to the specific circumstances.

[0040] The foregoing has provided a detailed description of a compound enzyme preparation apparatus and method, and its application in the efficient removal of dental plaque, as provided in the embodiments of this application. Specific examples have been used to illustrate the principles and implementation methods of this application. The descriptions of the above embodiments are only for the purpose of helping to understand the technical solutions and core ideas of this application. Those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. These modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of this application.

Claims

1. A compound enzyme preparation apparatus, comprising: The base is provided with two sets of support frames (2), and a connecting platform (21) is provided on the two side support frames (2). An extension platform (22) and an L-bracket (23) are also provided on the outside of the two side support frames (2). Fermentation tank (3), which is fixed at the middle position on the top surface of the connecting platform (21); Its characteristic is that it further includes: Adjustment structure (5), the adjustment structure (5) is provided in two sets and both are rotatably set between the side walls of the two side support frames (2), and the adjustment structure (5) holds a triangular bottle that can swing and adjust accordingly. The drainage structure (7) is slidably disposed on the extension platform (22), and both sides of the drainage structure (7) are provided with drainage tubes (731) that can be raised and lowered. The culture structure (6) is set on the L support (23). The culture structure (6) is connected to the fermenter (3) by a growth culture tube (33). The middle section of the growth culture tube (33) is also provided with a valve (661) for controlling the flow rate. The fermenter (3) is provided with a fixed tube (63) and a piston tube (64) that are adjusted by the dissolved oxygen content in the tank. The piston tube (64) slides along the inner wall of the fixed tube (63). A transmission component for controlling the rotation adjustment of the valve (661) is also provided between the piston tube (64) and the valve (661).

2. The compound enzyme preparation apparatus according to claim 1, characterized in that, A support column is provided on the connecting platform (21), and a pump body is provided on the top of the support column. An inoculation tube (31) is connected to both the inlet and outlet of the pump body. One end of one section of the inoculation tube (31) leads into the fermenter (3), and the other end of the inoculation tube (31) is a rigid tube. A lifting assembly (4) is installed on the base. The lifting end of the lifting assembly (4) is connected synchronously with the rigid tube part on the inoculation tube (31).

3. The compound enzyme preparation apparatus according to claim 2, characterized in that, The adjustment structure (5) includes a fixed frame (51), an adjustment frame (52), a rotating frame (55), and an output component. One end of the adjustment frame (52) slides between the inner walls of the fixed frame (51) through a threaded transmission. The adjustment frame (52) has a groove 2. The output component is set in the groove 2 and a motor 2 is connected to one side of it. Two fixed plates (54) are set on the adjustment frame (52). The output end of the output component is rotatably set between the opposite side walls of the two fixed plates (54). The output end of the output component is also connected synchronously with the rotating frame (55). The rotating frame (55) swings back and forth in the middle position of the two fixed plates (54). A limit shaft (554) is set between the rotating frame (55) and the fixed plate (54). The limit shaft (554) passes through the rotating frame (55) and is synchronous with it. The end of the limit shaft (554) moves between the inner walls of the fixed plate (54). Two meshing transmission gears (24) are rotatably arranged between the opposite side walls of the two side support frames (2). The transmission gears (24) are located above the fixed frame (51) and rotate coaxially with it. A motor is also connected below one of the transmission gears (24).

4. The compound enzyme preparation apparatus according to claim 3, characterized in that, The rotating frame (55) is arranged in a "U" shape. A heating element (553) is provided on the concave surface of the rotating frame (55). A U-shaped frame (551) is rotatably connected to both ends of the rotating frame (55) via a shaft. A "T"-shaped piston shaft is elastically provided at the middle position of the U-shaped frame (551). An arc-shaped positioning clamp (552) is connected to the end of the piston shaft facing the middle position of the rotating frame (55). Sponge pads are glued to the opposite side wall surfaces of the positioning clamps (552) on both sides.

5. The compound enzyme preparation apparatus according to claim 1, characterized in that, The drainage structure (7) includes a sliding seat (71), a lifting platform (72), and a drainage pipe (731). The sliding seat (71) is slidably adjusted along the length of the extension platform (22). A lifting cylinder is provided on the top surface of the sliding seat (71). The lifting platform (72) is installed on the output shaft end of the lifting cylinder. A second pump body (73) is also provided on the lifting platform (72). The drainage pipe (731) is connected to both the inlet and outlet of the second pump body (73).

6. The compound enzyme preparation apparatus according to claim 1, characterized in that, The culture structure (6) includes a fixed tube (63), a piston tube (64), and an adjusting gear (65). One end of the fixed tube (63) is installed on the side wall of the fermenter (3) and extends into its interior. One end of the piston tube (64) always slides between the inner walls of the fixed tube (63). A connecting bolt is provided on the fixed tube (63), and a connecting bolt is provided on the piston tube (64). A tension spring is provided between the connecting bolt and the connecting bolt, and several toothed grooves are opened on the outer surface of the piston tube (64) along its length direction.

7. The compound enzyme preparation apparatus according to claim 6, characterized in that, The transmission assembly includes an adjusting gear one (65), an adjusting gear two (66), a gear condition one (621), and a gear condition two (627). The adjusting gear one (65) is rotatably mounted on the L-bracket (23) via a shaft connection, and the adjusting gear one (65) is meshed with a tooth groove for transmission. A guide rail platform (62) is also provided on one side of the adjusting gear one (65) on the L-bracket (23). A guide bar two is slidably mounted between the inner walls of the guide rail platform (62) along its length direction. The gear condition one (621) is fixed to the top surface of the guide bar two. An iron positioning block (625) is provided on the side wall of the upper tooth condition one (621). A rack block (622) is also elastically provided on one side of the tooth condition one (621). Two "L"-shaped connecting brackets (626) are also provided on the top surface of the tooth condition one (621). The tooth condition two (627) is fixed between the connecting brackets (626) on both sides. The adjusting gear two (66) is located above the valve disc (661) and coaxially connected to it. The adjusting gear two (66) and the tooth condition two (627) are meshed and driven.

8. The compound enzyme preparation apparatus according to claim 7, characterized in that, The top surface of the guide rail platform (62) is provided with a limiting seat (624). The limiting seat (624) has a positioning groove matching the positioning block (625) on the side wall facing the positioning block (625). The inner wall of the positioning groove is embedded with a silicon steel sheet, and an external power supply electrically connected to the silicon steel sheet is installed on the side wall of the guide rail platform (62). A connecting block 2 is provided on the side wall of the gear condition 1 (621), and a connecting block 3 is provided on the end wall of the rack block (622). An "I"-shaped piston rod (623) passes through the connecting block 2 and the connecting block 3, and a spring is sleeved on the piston rod (623).

9. A method for preparing a complex enzyme using the complex enzyme preparation apparatus according to claim 1, characterized in that, Includes the following steps: S1. Activation culture: Prepare YPD medium and put it in an Erlenmeyer flask. After autoclaving and cooling the medium to room temperature, pick 1 loop of bacteria from the recombinant Pichia pastoris slant medium in a clean bench and inoculate it into the YPD medium. Place it on one of the adjustment structures (5) and fix it. Set the heating temperature to 30±2℃ and the rotation speed to 200±5 r / min. Activate the medium for 24 hours. S2, bacterial culture: BMGY culture medium is prepared and placed in another triangular flask. After autoclaving and cooling the culture medium to room temperature, the triangular flask is fixed on another regulating structure (5). The activated bacterial solution is transferred to the triangular flask through the drainage structure (7). The heating temperature is controlled at 30±2℃, the rotation speed is set at 200±5 r / min, and the culture is continued for 24 hours. S3, High-density culture: BSM culture was prepared in fermenter (3). The pH electrode and dissolved oxygen electrode were calibrated and assembled using calibration solution. Finally, the electrode protective sleeve was wrapped with kraft paper and sterilized at 115±5℃ for 20min. After the culture medium cooled to room temperature, the BMGY bacterial solution in S2 was inoculated into the BSM culture medium at a 10% inoculation rate. The aeration rate was adjusted to keep the dissolved oxygen above 25%. The temperature was set at 30±2℃ and the rotation speed was 500±10 r / min. When the dissolved oxygen rose rapidly, the feed culture medium was started to be added to the fermenter (3). When the bacterial concentration reached the set value, the feed culture medium pump was turned off. After the Pichia pastoris consumed all the glycerol in the culture medium and maintained a substrate-deficient state for 1 hour, the induction temperature was set and the induction culture medium was started to be added to induce the expression of the complex enzyme, dextranase.

10. The application of a composite enzyme obtained by the method for preparing a composite enzyme according to claim 9 in the efficient removal of dental plaque, characterized in that, The complex enzyme can remove biofilm in the oral cavity.

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