A kind of androstenedione preparation device and preparation method

Abstract

The invention belongs to the technical field of bioengineering equipment, and specifically is an androstenedione preparation device and preparation method, comprising a base; a rotating seat is fixedly connected to the top surface of the base; a support column is fixedly connected to the top surface of the rotating seat at the middle position; a plurality of uniformly arranged support arms are fixedly connected to the surface of the support column; the ends of the support arms away from the rotating seat are rotatably connected to a fermenter; a sealing cover is installed at the opening of the fermenter; a feed pipe and an exhaust pipe are fixedly connected to the end of the sealing cover away from the fermenter; a driving rod is rotatably connected to the side of the sealing cover close to the fermenter; blades are arranged on the outer side of the driving rod, and the fermenter is driven to rotate by the rotating seat, so that the fermented liquid can complete the centrifugal operation in the fermenter, thereby avoiding multiple extraction and transportation of the fermented liquid, thereby reducing the probability of the fermented liquid being contaminated by the outside, and at the same time reducing the work process and improving the production efficiency.

Description

Technical Field

[0001] The invention belongs to the technical field of bioengineering equipment, and in particular relates to a device and method for preparing androstenedione. Background Art

[0002] Steroid drugs have high value in clinical medical treatment and can be used in chronic diseases such as cardiovascular disease, hypertension, and tumors. They can also be used in neurological diseases such as depression and anxiety. With the change of production technology, 4-androstenedione, an important starting material, is mostly produced by microbial conversion and fermentation of progesterone.

[0003] The preparation of 4-androstenedione in the prior art is generally prepared by microbial fermentation. First, the bacteria and progesterone are introduced into a fermentation tank, and then the fermentation is carried out under constant temperature stirring. After the fermentation is completed, the fermentation liquid is extracted and centrifuged, and finally the supernatant is extracted and purified to obtain the final product.

[0004] In the above technology, after the fermentation operation is completed in the fermentation tank, the fermentation liquid in the fermentation tank needs to be extracted, and then the fermentation liquid is introduced into a centrifuge, and finally the supernatant is separated by centrifugation. Since it involves multiple liquid transfers, it is very easy to cause contamination of the raw materials. At the same time, the transportation of the liquid also adds more operating procedures, thereby reducing production efficiency.

[0005] To this end, the present invention provides a device and method for preparing androstenedione. Summary of the invention

[0006] In order to make up for the deficiencies of the prior art, at least one technical problem raised in the background technology is solved.

[0007] The technical solution adopted by the present invention to solve its technical problems is as follows: an androstenedione preparation device described in the present invention comprises a base; a rotating seat is fixedly connected to the top surface of the base, and the rotating seat is driven by a servo motor; a support column is fixedly connected to the top surface of the rotating seat at a middle position; a plurality of uniformly arranged support arms are fixedly connected to the surface of the support column, and the number of the support arms is an even number; one end of the support arm away from the rotating seat is rotatably connected to a fermentation tank; a heating component is installed on the surface of the fermentation tank; the heating component heats the fermentation tank through an electric heating wire; a sealing cover is installed at the opening of the fermentation tank; a feed pipe and an exhaust pipe are fixedly connected to the end of the sealing cover away from the fermentation tank, and a valve is installed inside the exhaust pipe; a driving rod is rotatably connected to the side of the sealing cover close to the fermentation tank, and is driven by a servo motor; blades are arranged on the outside of the driving rod.

[0008] Preferably, a through hole is provided on the top surface of the rotating seat, and the through hole passes through the rotating seat; a hydraulic rod is fixedly connected to the top surface of the base at the through hole; the telescopic end of the hydraulic rod is fixedly connected to a temporary storage tank; a plurality of evenly arranged support frames are fixedly connected to the bottom of the temporary storage tank, and the support frames and support arms are arranged in a one-to-one correspondence; a discharge pipe is fixedly connected to one end of the support frame away from the hydraulic rod; the positions of the discharge pipe and the feed pipe correspond, and the diameter of the discharge pipe is smaller than the diameter of the feed pipe; a conduit is fixedly connected between the discharge pipe and the temporary storage tank, and an electric control valve is provided at the connection between the conduit and the temporary storage tank.

[0009] Preferably, a bellows is fixedly connected to the top surface of the rotating seat between two adjacent support arms; the diameter of the top of the bellows is larger than the diameter of the discharge pipe and corresponds to the position of the discharge pipe; sewage outlets are provided on the side surface of the rotating seat at the corresponding position of the bellows, and the bottom of the bellows is connected to the sewage outlet; a liquid guide ring is installed on the side surface of the rotating seat, and one side of the liquid guide ring is higher than the other side; a notch is provided at the lowest end of the liquid guide ring.

[0010] Preferably, a telescopic rod is fixedly connected to the bottom of the liquid guide ring; the telescopic rod is fixed by a latch; and a baffle is fixedly connected to the outer side of the liquid guide ring.

[0011] Preferably, an internal gear is rotatably connected to the top surface of the rotating seat and the outer side of the support column; a plurality of sub-gears are meshingly connected to the inner side of the internal gear, and the sub-gears are rotatably connected to the top surface of the rotating seat; a threaded rod is fixedly connected to the top surface of the sub-gear; a movable plate is threadedly connected to the surface of the threaded rod; one end of the movable plate is slidably connected to the surface of the support column; and the end of the movable plate away from the support column is fixedly connected to the top surface of the bellows.

[0012] Preferably, the surface of the driving rod is slidably connected to a sleeve rod; the blades are fixedly connected to the surface of the sleeve rod; the end of the sleeve rod close to the sealing cover is rotatably connected to a rotating ring; a connecting plate is fixedly connected to one side of the rotating ring; a lifting rod is fixedly connected to the end of the connecting plate away from the rotating ring, and the lifting rod is located at the feed pipe; a closing cover is threadedly connected to the feed hopper; a socket is provided in the middle of the closing cover, and the socket is adapted to the lifting rod; a pair of symmetrically arranged receiving grooves are provided on the surface of the lifting rod; and a clamping plate is rotatably connected to the end of the receiving groove away from the connecting plate.

[0013] Preferably, a rubber plug is provided on one side of the closing cover close to the feed pipe at a position corresponding to the plug hole, and the diameter of the rubber plug close to the plug hole is smaller than that of the plug hole; an elastic rod is fixedly connected between the rubber plug and the closing cover; the diameter of the rubber plug close to the plug hole is smaller than the diameter of the end away from the plug hole.

[0014] Preferably, a plurality of evenly arranged hydraulic cylinders are fixedly connected around the base; the telescopic ends of the hydraulic cylinders are commonly fixedly connected with a protective ring; the end of the protective ring close to the support column is rotatably connected with a plurality of evenly arranged rollers; and a connecting ring is commonly installed at the end of the plurality of support arms away from the support column.

[0015] Preferably, a method for preparing androstenedione, the method adopts an androstenedione preparation device described in any one of the above, comprising the following steps:

[0016] S1. Take the strain, put the frozen glycerol tube of Pseudomonas alkylphenolica GR-4-4 into an ice box to thaw, inoculate it into the LB plate with an inoculation loop on an ultra-clear workbench, and culture it in a 28°C incubator for 23-24 hours. Then pick a single colony with an inoculation loop and inoculate it again into the LB plate, and culture it in a 28°C incubator for 23-24 hours.

[0017] S2. Culture the strains. Use SX liquid culture medium, prepare 100 ml SX liquid culture medium in a 250 ml conical flask, and sterilize at 121°C for 30-35 min. Take the strain Pseudomonas alkylphenolica GR-4-4 plate, pick a single colony with an inoculation loop and inoculate it into the SX liquid culture medium, and then culture it in a constant temperature shaker at 28°C and 180 rpm for 15-16 h.

[0018] S3, fermentation culture, 9.5-10.5L of SX liquid culture medium is introduced into the fermenter, sterilized at 121°C for 30-35min, and then the fermentation liquid is cooled to room temperature, the cultured strain Pseudomonas alkylphenolica GR-4-4 seed liquid is aseptically poured into the fermenter, and the servo motor is started to drive the driving rod to rotate, and the driving rod drives the sleeve rod and the blades thereon to stir the liquid in the fermenter, and the constant temperature of 28°C is maintained for fermentation for 71.5-72h.

[0019] S4, extracting the transformant, adding alkali to the fermentation liquid in the fermentation tank to adjust the pH to 11.8-12, then heating the fermentation tank, boiling the fermentation liquid in the fermentation tank to inactivate the bacteria, then the rotating seat drives the support arm and the fermentation tank thereon to rotate, and then the liquid in the fermentation tank is stratified by centrifugal action, and then the supernatant in the fermentation tank is collected.

[0020] S5. The transformed product is initially purified, and the supernatant is concentrated to 0.9-1L in a sand bath pot. After the supernatant is cooled to room temperature, 29%-30% ethyl acetate is added to the supernatant, and the pH value is adjusted to 2-3. The temperature is controlled at 49-50°C and stirred for extraction for 5-10 minutes. The mixture is then placed in a separatory funnel and allowed to stand for separation, and the aqueous phase is collected. After repeating the extraction three times, the extracts are combined and the solvent is recovered on a rotary evaporator to 5-10ml, and then dried in a vacuum drying oven.

[0021] S6. The transformed product is further purified by placing the dried product in a beaker, adding 100 times of water, adding 19% to 20% of sodium hydroxide, boiling and reacting for 29-30 minutes, cooling and adding water to 50 times, adjusting the pH value to 2-3 with hydrochloric acid at low temperature, heating and dehydrating, filtering, washing with water to a pH value of 5-6, and drying at 50°C to obtain the extracted product.

[0022] Preferably, the fermenter in S4 needs to be deflected before centrifugal separation so that the fermenter is no longer perpendicular to the support arm, making the stratification more obvious. At the same time, it is also necessary to disconnect the fermenter from the external pipeline to avoid the pipeline from being entangled when the support arm drives the fermenter to rotate.

[0023] The beneficial effects of the present invention are as follows:

[0024] 1. The androstenedione preparation device and preparation method described in the present invention drive the fermentation tank to rotate by a rotating seat, so that the fermented liquid can complete the centrifugal operation in the fermentation tank, thereby avoiding multiple extraction and transportation of the fermentation liquid, thereby reducing the probability of the fermentation liquid being contaminated by the outside world, and at the same time reducing the work flow and improving the production efficiency.

[0025] 2. The androstenedione preparation device and preparation method described in the present invention receive and drain sewage through a bellows and a liquid guide ring, which can prevent sewage from flowing around, reduce the difficulty of users cleaning the embodiment of the present invention, and improve the cleaning efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] The present invention will be further described below in conjunction with the accompanying drawings.

[0027] Figure 1 is a stereogram of the present invention;

[0028] Figure 2 It is a structural schematic diagram of the support column of the present invention;

[0029] Figure 3 It is a structural schematic diagram of the hydraulic rod in the present invention;

[0030] Figure 4 It is a schematic diagram of the structure of the fermentation tank in the present invention;

[0031] Figure 5 is a cross-sectional view of a fermentation tank in the present invention;

[0032] Figure 6 is a cross-sectional view of the closure cover of the present invention;

[0033] Figure 7 It is a schematic diagram of the structure of the guard ring in the present invention;

[0034] Figure 8 is a flow chart of the method of the present invention;

[0035] In the figure: 1. base; 2. rotating seat; 3. support column; 4. support arm; 5. fermentation tank; 6. sealing cover; 7. feed pipe; 8. exhaust pipe; 9. drive rod; 10. blade; 11. through hole; 12. hydraulic rod; 13. temporary storage tank; 14. support frame; 15. feed pipe; 16. conduit; 17. bellows; 18. sewage outlet; 19. liquid guide ring; 20. notch; 21. telescopic rod; 22. baffle; 23. internal gear; 24. sub gear; 25. threaded rod; 26. moving plate; 27. sleeve rod; 28. rotating ring; 29. ​​connecting plate; 30. lifting rod; 31. closing cover; 32. jack; 33. receiving groove; 34. card plate; 35. rubber plug; 36. elastic rod; 37. hydraulic cylinder; 38. protective ring; 39. roller; 40. connecting ring. DETAILED DESCRIPTION

[0036] In order to make the technical means, creative features, objectives and effects achieved by the present invention easy to understand, the present invention is further explained below in conjunction with specific implementation methods.

[0037] like Figures 1 to 4As shown, an androstenedione preparation device according to an embodiment of the present invention comprises a base 1; a rotating seat 2 is fixedly connected to the top surface of the base 1, and the rotating seat 2 is driven by a servo motor; a support column 3 is fixedly connected to the top surface of the rotating seat 2 at the middle position; a plurality of uniformly arranged support arms 4 are fixedly connected to the surface of the support column 3, and the number of the support arms 4 is an even number; a fermentation tank 5 is rotatably connected to one end of the support arm 4 away from the rotating seat 2; a heating component is installed on the surface of the fermentation tank 5; the heating component heats the fermentation tank 5 through an electric heating wire; The opening of the fermentation tank 5 is equipped with a sealing cover 6; the end of the sealing cover 6 away from the fermentation tank 5 is fixedly connected with a feed pipe 7 and an exhaust pipe 8, and a valve is installed inside the exhaust pipe 8; the side of the sealing cover 6 close to the fermentation tank 5 is rotatably connected with a driving rod 9, and is driven by a servo motor; the outer side of the driving rod 9 is provided with a blade 10; when working, in order to reduce the number of liquid transfers in the preparation process of androstenedione, the embodiment of the present invention can be used. First, the user introduces the culture solution and the strain into the fermentation tank 5 through the feed pipe 7, and then the servo motor is driven. The motor drives the driving rod 9 and the blade 10 to rotate, thereby stirring the liquid in the fermentation tank 5, so that the strain and the culture liquid are fully mixed. During the culture period, the user can also open the valve at the exhaust pipe 8 at regular intervals to ensure that the pressure inside the fermentation tank 5 is normal. When the fermentation is completed, the user needs to deflect the fermentation tank 5 so that the sealing cover 6 is close to the support arm 4. At this time, the fermentation tank 5 is no longer perpendicular to the support arm 4, but forms an acute angle with the support arm 4. Then the user drives the rotating seat 2 to rotate, and the rotating seat 2 drives the support column 3 and the support arm 4 to rotate, thereby driving the fermentation tank 5 on the support arm 4 to rotate together, thereby completing the centrifugal operation of the liquid in the fermentation tank 5, so that the liquid in the fermentation tank 5 is stratified, and then the user can extract the required supernatant and carry out the next purification operation. The fermentation tank 5 is driven to rotate by the rotating seat 2, so that the fermented liquid can complete the centrifugal operation in the fermentation tank 5, thereby avoiding multiple extractions and transportations of the fermentation liquid, thereby reducing the probability of the fermentation liquid being contaminated by the outside world, while reducing the work process and improving production efficiency.

[0038] like Figures 2 to 3As shown, a through hole 11 is provided on the top surface of the rotating seat 2, and the through hole 11 passes through the rotating seat 2; a hydraulic rod 12 is fixedly connected to the top surface of the base 1 at the through hole 11; a temporary storage tank 13 is fixedly connected to the telescopic end of the hydraulic rod 12; a plurality of evenly arranged support frames 14 are fixedly connected to the bottom of the temporary storage tank 13, and the support frames 14 and the support arms 4 are arranged in a one-to-one correspondence; a feed pipe 15 is fixedly connected to one end of the support frame 14 away from the hydraulic rod 12; the feed pipe 15 corresponds to the position of the feed pipe 7, and the diameter of the feed pipe 15 is smaller than the diameter of the feed pipe 7; a conduit 16 is fixedly connected between the feed pipe 15 and the temporary storage tank 13, and the connection between the conduit 16 and the temporary storage tank 13 An electric control valve is provided at the temporary storage tank 13; during operation, when the user needs to inject the culture solution or bacteria into the fermentation tank 5, the user needs to first inject the culture solution or bacteria into the temporary storage tank 13, and then the user drives the temporary storage tank 13 to descend through the hydraulic rod 12 in the through hole 11, and then the temporary storage tank 13 drives the support frame 14 to descend until the discharge pipe 15 at the end of the temporary storage tank 13 is inserted into the feed pipe 7 on the surface of the sealing cover 6, and then the user can open the electric control valve at the conduit 16 to introduce the culture solution or bacteria in the temporary storage tank 13 into the fermentation tank 5 through the feed pipe 7, thereby realizing the simultaneous filling of multiple fermentation tanks 5, thereby shortening the filling time and improving the filling efficiency of the fermentation tank 5.

[0039] like Figures 1 to 3 As shown, a bellows 17 is fixedly connected between two adjacent support arms 4 on the top surface of the rotating seat 2; the diameter of the top of the bellows 17 is larger than the diameter of the discharge pipe 15, and corresponds to the position of the discharge pipe 15; a sewage outlet 18 is provided on the side surface of the rotating seat 2 at the corresponding position of the bellows 17, and the bottom of the bellows 17 is connected to the sewage outlet 18; a liquid guide ring 19 is installed on the side surface of the rotating seat 2, and one side of the liquid guide ring 19 is higher than the other side; a notch 20 is provided at the lowest end of the liquid guide ring 19; during operation, when the bacteria or culture solution in the temporary storage tank 13 is filled, in order to avoid residual bacteria or culture solution in the temporary storage tank 13, the conduit 16 and the discharge pipe 15, which may cause contamination inside them, they need to be cleaned, and for this purpose, the user needs to first clean the bellows 19. The top of the bellows 17 is butted against the discharge pipe 15, and then the user injects clean water into the temporary storage tank 13, which then guides the clean water into each conduit 16 and finally discharges it from the discharge pipe 15 at the end of the conduit 16, thereby achieving the purpose of cleaning the temporary storage tank 13, the conduit 16 and the discharge pipe 15. At the same time, the cleaned waste water can also flow out from the sewage outlet 18 on the surface of the rotating seat 2 toilet under the guidance of the bellows 17, and finally flow to the surface of the liquid guide ring 19. Since the liquid guide ring 19 is inclined, the sewage on the surface of the liquid guide ring 19 will flow to the gap 20 and finally be discharged from the gap 20. The sewage is received and drained by the bellows 17 and the liquid guide ring 19, which can prevent the sewage from flowing everywhere, and can also reduce the difficulty of the user to clean the embodiment of the present invention and improve the cleaning efficiency.

[0040] like Figures 1 to 3 As shown, a telescopic rod 21 is fixedly connected to the bottom of the liquid guide ring 19; the telescopic rod 21 is fixed by a latch; a baffle 22 is fixedly connected to the outer side of the liquid guide ring 19; during operation, after the temporary storage tank 13, the guide tube 16 and the discharge tube 15 are cleaned, the user needs to release the fixation of the telescopic rod 21 through the latch, and enable the telescopic rod 21 to drive the liquid guide ring 19 to descend, so that the inner side of the liquid guide ring 19 is no longer in contact with the surface of the rotating seat 2, thereby preventing the surface of the rotating seat 2 from contacting the guide ring 19 when the rotating seat 2 rotates. The liquid ring 19 continues to rub, causing the liquid guide ring 19 to wear and affecting its service life. At the same time, after the liquid guide ring 19 drops, the baffle 22 on its outer side can still cover the sewage outlet 18. Therefore, when the rotating seat 2 rotates, the sewage in the sewage outlet 18 is thrown out due to the centrifugal effect. The baffle 22 can catch the thrown sewage and collect the sewage and then guide it out from the gap 20, thereby avoiding the residual sewage in the sewage outlet 18 being directly thrown into the surrounding environment when the rotating seat 2 rotates, causing pollution to the surrounding environment.

[0041] like Figure 3 As shown, the top surface of the rotating seat 2 is rotatably connected with the outer side of the support column 3 by an internal gear 23; the inner side of the internal gear 23 is meshed with a plurality of sub-gears 24, and the sub-gears 24 are rotatably connected with the top surface of the rotating seat 2; the top surface of the sub-gear 24 is fixedly connected with a threaded rod 25; the surface of the threaded rod 25 is threadedly connected with a movable plate 26; one end of the movable plate 26 is slidably connected with the surface of the support column 3; the end of the movable plate 26 away from the support column 3 is fixedly connected with the top surface of the bellows 17; during operation, when the user needs to dock the bellows 17 with the discharge pipe 15, the user only needs to screw one of the plurality of threaded rods 25, and the screwed threaded rod 25 can drive the bottom sub-gear 24 to cooperate with the internal gear 23 to ... Its internal gear 23 and threaded rod 25 rotate, and the rotation of the threaded rod 25 can drive the movable plate 26 to rise, and then drive the bellows 17 to rise, so that the top of the bellows 17 is docked with the discharge pipe 15. Therefore, only one threaded rod 25 needs to be rotated to simultaneously drive multiple bellows 17 to complete docking with the discharge pipe 15, thereby reducing the difficulty of operation for users. At the same time, when the bellows 17 is not in use, the user can screw the threaded rod 25 to make the movable plate 26 drive the top of the bellows 17 to descend, thereby compressing the bellows 17, and making it convenient for the user to store the bellows 17. The traction of the bellows 17 by the movable plate 26 can also prevent the bellows 17 from swinging randomly due to the centrifugal effect when the rotating seat 2 rotates.

[0042] like Figures 4 to 5As shown, the surface of the driving rod 9 is slidably connected with a sleeve rod 27; the blade 10 is fixedly connected to the surface of the sleeve rod 27; the end of the sleeve rod 27 close to the sealing cover 6 is rotatably connected with a rotating ring 28; one side of the rotating ring 28 is fixedly connected with a connecting plate 29; the end of the connecting plate 29 away from the rotating ring 28 is fixedly connected with a lifting rod 30, and the lifting rod 30 is located at the feeding pipe 7; the feeding hopper is threadedly connected with a closing cover 31; the middle part of the closing cover 31 is provided with a plug hole 32, and the plug hole 32 is adapted to the lifting rod 30; the surface of the lifting rod 30 is provided with a pair of symmetrically arranged receiving grooves 33; the end of the receiving groove 33 away from the connecting plate 29 is rotatably connected with a clamping plate 34; during operation, when the servo motor drives the driving rod 9 to rotate, the driving rod 9 will drive the sleeve rod 27 to rotate, and then drive the blades 1 on the surface of the sleeve rod 27 0 rotates, thereby completing the stirring of the liquid in the fermentation tank 5. When the liquid in the fermentation tank 5 needs to be centrifuged, the user needs to pull the lifting rod 30 out of the feed port. During this process, the lifting rod 30 will lift the sleeve rod 27 through the rotating ring 28, thereby driving the sleeve rod 27 and the blades 10 away from the liquid in the fermentation tank 5, thereby preventing the sleeve rod 27 and the blades 10 from being inserted into the liquid, resulting in obstruction of the stratification of the liquid. To fix the lifting rod 30, it is necessary to align the insertion hole 32 at the end of the closing cover 31 with the lifting rod 30 so that the lifting rod 30 can pass through the insertion hole 32, and then screw the closing cover 31 onto the feeding pipe 7. Thereafter, the user rotates the card plate 34 in the receiving groove 33 to make the card plate 34 rotate out of the receiving groove 33. At this time, the card plate 34 can play a limiting role to prevent the lifting rod 30 from falling back into the feeding pipe 7 and the inside of the fermentation tank 5.

[0043] like Figure 5 to Figure 6 As shown, a rubber plug 35 is provided at a position corresponding to the insertion hole 32 on one side of the closing cover 31 close to the feeding pipe 7, and the diameter of the rubber plug 35 at one end close to the insertion hole 32 is smaller than the insertion hole 32; an elastic rod 36 is fixedly connected between the rubber plug 35 and the closing cover 31; the diameter of the rubber plug 35 at one end close to the insertion hole 32 is smaller than the diameter of the end away from the insertion hole 32; during operation, when the fermentation tank 5 needs to be sealed and fermented, the user needs to insert the rubber plug 35 into the insertion hole 32 to seal the insertion hole 32, and then screw the closing cover 31 on the feeding pipe 7, at this time, the closing cover 31 can complete the sealing of the feeding pipe 7, and when the lifting rod 30 needs to pass through the insertion hole 32, the user only needs to pull out the rubber plug 35 at the insertion hole 32, so that the lifting rod 30 can pass through the insertion hole 32 smoothly, and at the same time, the elastic rod 36 can also limit the rubber plug 35 in the closing cover 31, thereby avoiding the loss of the rubber plug 35.

[0044] like Figure 1 and Figure 7As shown, a plurality of hydraulic cylinders 37 are fixedly connected to the four sides of the base 1 evenly arranged; the telescopic ends of the hydraulic cylinders 37 are commonly fixedly connected with a protective ring 38; the end of the protective ring 38 close to the support column 3 is rotatably connected with a plurality of rollers 39 evenly arranged; a plurality of support arms 4 are commonly installed with a connecting ring 40 at one end away from the support column 3; during operation, when the fermentation tank 5 needs to be centrifuged, the hydraulic cylinder 37 will drive the protective ring 38 to rise, and make the rollers 39 on the inner side of the protective ring 38 abut against the surface of the connecting ring 40, so that when the support arm 4 and the connecting ring 40 rotate, the connecting ring 40 provides centripetal force for each support arm 4, so as to avoid the support arm 4 from breaking due to centrifugal action at high speed, thereby causing the support column 3 to be thrown out and cause harm to the staff.

[0045] like Figure 8 As shown, a method for preparing androstenedione, the method adopts an androstenedione preparation device described in any one of the above, comprising the following steps:

[0046] S1. Take the strain, put the frozen glycerol tube of Pseudomonas alkylphenolica GR-4-4 into an ice box to thaw, inoculate it into the LB plate with an inoculation loop on an ultra-clear workbench, and culture it in a 28°C incubator for 23-24 hours. Then pick a single colony with an inoculation loop and inoculate it again into the LB plate, and culture it in a 28°C incubator for 23-24 hours.

[0047] S2. Culture the strains. Use SX liquid culture medium, prepare 100 ml SX liquid culture medium in a 250 ml conical flask, and sterilize at 121°C for 30-35 min. Take the strain Pseudomonas alkylphenolica GR-4-4 plate, pick a single colony with an inoculation loop and inoculate it into the SX liquid culture medium, and then culture it in a constant temperature shaker at 28°C and 180 rpm for 15-16 h.

[0048] S3, fermentation culture, 9.5-10.5L of SX liquid culture medium is introduced into the fermentation tank 5, sterilized at 121°C for 30-35min, and then the fermentation liquid is cooled to room temperature, and the cultured strain Pseudomonas alkylphenolica GR-4-4 seed liquid is aseptically poured into the fermentation tank 5, and the servo motor is started to drive the driving rod 9 to rotate, and the driving rod 9 drives the sleeve rod 27 and the blades 10 thereon to stir the liquid in the fermentation tank 5, and the constant temperature of 28°C is maintained for fermentation for 71.5-72h.

[0049] S4, extracting the transformant, adding alkali to the fermentation liquid in the fermentation tank 5 to adjust the pH to 11.8-12, then heating the fermentation tank 5, boiling the fermentation liquid in the fermentation tank 5 to inactivate the bacteria, then rotating the support arm 4 and the fermentation tank 5 thereon to rotate, and then the liquid in the fermentation tank 5 is stratified by centrifugal action, and then the supernatant in the fermentation tank 5 is collected.

[0050] S5. The transformed product is initially purified, and the supernatant is concentrated to 0.9-1L in a sand bath pot. After the supernatant is cooled to room temperature, 29%-30% ethyl acetate is added to the supernatant, and the pH value is adjusted to 2-3. The temperature is controlled at 49-50°C and stirred for extraction for 5-10 minutes. The mixture is then placed in a separatory funnel and allowed to stand for separation, and the aqueous phase is collected. After repeating the extraction three times, the extracts are combined and the solvent is recovered on a rotary evaporator to 5-10ml, and then dried in a vacuum drying oven.

[0051] S6. The transformed product is further purified by placing the dried product in a beaker, adding 100 times of water, adding 19% to 20% of sodium hydroxide, boiling and reacting for 29-30 minutes, cooling and adding water to 50 times, adjusting the pH value to 2-3 with hydrochloric acid at low temperature, heating and dehydrating, filtering, washing with water to a pH value of 5-6, and drying at 50°C to obtain the extracted product.

[0052] The fermentation tank 5 in S4 is in the process of centrifugal separation. Before centrifugal separation, it is necessary to deflect the fermentation tank 5 so that the fermentation tank 5 is no longer perpendicular to the support arm 4, so that the stratification is more obvious. At the same time, it is also necessary to disconnect the fermentation tank 5 from the external pipeline to avoid the pipeline being entangled when the support arm 4 drives the fermentation tank 5 to rotate.

[0053] During operation, in order to reduce the number of liquid transfers during the preparation of androstenedione, the embodiment of the present invention can be used. First, the user introduces the culture solution and the strain into the fermentation tank 5 through the feed pipe 7, and then the servo motor drives the drive rod 9 and the blade 10 to rotate, thereby stirring the liquid in the fermentation tank 5 so that the strain and the culture solution are fully mixed. During the culture period, the user can also open the valve at the exhaust pipe 8 at regular intervals to ensure that the pressure inside the fermentation tank 5 is normal. When the fermentation is completed, the user needs to deflect the fermentation tank 5 so that the sealing cover 6 is close to the support arm 4. At this time, the fermentation tank 5 is no longer perpendicular to the support arm 4, but forms a sharp gap with the support arm 4. Angle, and then the user drives the rotating seat 2 to rotate, and the rotating seat 2 drives the support column 3 and the support arm 4 to rotate, and then drives the fermentation tank 5 on the support arm 4 to rotate together, thereby completing the centrifugal operation of the liquid in the fermentation tank 5, so that the liquid in the fermentation tank 5 is stratified, and then the user can extract the required supernatant to carry out the next purification operation, and drive the fermentation tank 5 to rotate by the rotating seat 2, so that the fermented liquid can complete the centrifugal operation in the fermentation tank 5, thereby avoiding multiple extractions and transportations of the fermentation liquid, thereby reducing the probability of the fermentation liquid being contaminated by the outside world, while reducing the work process and improving production efficiency.

[0054] When the user needs to inject the culture solution or bacteria into the fermentation tank 5, the user needs to first inject the culture solution or bacteria into the temporary storage tank 13, and then the user drives the temporary storage tank 13 to descend through the hydraulic rod 12 in the through hole 11, and then the temporary storage tank 13 drives the support frame 14 to descend until the discharge pipe 15 at the end of the temporary storage tank 13 is inserted into the feed pipe 7 on the surface of the sealing cover 6, and then the user can open the electric control valve at the conduit 16 to introduce the culture solution or bacteria in the temporary storage tank 13 into the fermentation tank 5 through the feed pipe 7, thereby realizing the simultaneous filling of multiple fermentation tanks 5, thereby shortening the filling time and improving the filling efficiency of the fermentation tank 5.

[0055] After the bacteria or culture solution in the temporary storage tank 13 is filled, in order to avoid residual bacteria or culture solution in the temporary storage tank 13, the conduit 16 and the discharge pipe 15, which may cause internal contamination, they need to be cleaned. For this purpose, the user needs to first connect the top of the bellows 17 with the discharge pipe 15, and then the user injects clean water into the temporary storage tank 13, and then the temporary storage tank 13 guides the clean water into each conduit 16, and finally discharges it from the discharge pipe 15 at the end of the conduit 16, thereby cleaning the temporary storage tank 13, the conduit 16 and the discharge pipe 15. 6 and the discharge pipe 15, and the cleaned waste water can also flow out from the sewage outlet 18 on the surface of the rotating seat 2 under the guidance of the bellows 17, and finally flow to the surface of the liquid guide ring 19. Since the liquid guide ring 19 is inclined, the sewage on the surface of the liquid guide ring 19 will flow to the gap 20 and finally be discharged from the gap 20. The sewage is received and drained by the bellows 17 and the liquid guide ring 19, which can prevent the sewage from flowing around, and can also reduce the difficulty of the user to clean the embodiment of the present invention and improve the cleaning efficiency.

[0056] After the temporary storage tank 13, the conduit 16 and the discharge pipe 15 are cleaned, the user needs to release the fixation of the telescopic rod 21 through the latch, and enable the telescopic rod 21 to drive the liquid guide ring 19 to descend, so that the inner side of the liquid guide ring 19 is no longer in contact with the surface of the rotating seat 2, thereby preventing the surface of the rotating seat 2 from continuously rubbing against the liquid guide ring 19 when the rotating seat 2 rotates, causing the liquid guide ring 19 to wear and affect its service life. At the same time, after the liquid guide ring 19 descends, the baffle 22 on its outer side can still cover the sewage outlet 18, so when the rotating seat 2 rotates, the sewage in the sewage outlet 18 is thrown out due to the centrifugal effect, and the baffle 22 can catch the thrown sewage and collect the sewage and then lead it out from the gap 20, thereby preventing the residual sewage in the sewage outlet 18 from being directly thrown into the surrounding environment when the rotating seat 2 rotates, causing pollution to the surrounding environment.

[0057] When the user needs to dock the bellows 17 with the discharge tube 15, the user only needs to screw one of the multiple threaded rods 25, and the screwed threaded rod 25 can drive the other internal gears 23 and the threaded rod 25 to rotate through the cooperation between its bottom sub-gear 24 and the internal gear 23, and the rotation of the threaded rod 25 can drive the movable plate 26 to rise, and then drive the bellows 17 to rise, so that the top of the bellows 17 is docked with the discharge tube 15. Therefore, only one threaded rod 25 needs to be rotated to simultaneously drive multiple bellows 17 to complete the docking with the discharge tube 15, thereby reducing the difficulty of operation for the user. At the same time, when the bellows 17 is not in use, the user screws the threaded rod 25 to make the movable plate 26 drive the top of the bellows 17 to descend, thereby compressing the bellows 17, and then facilitating the user to store the bellows 17. The traction of the movable plate 26 on the bellows 17 can also prevent the bellows 17 from swinging randomly due to the centrifugal force when the rotating seat 2 rotates.

[0058] When the servo motor drives the driving rod 9 to rotate, the driving rod 9 will drive the sleeve rod 27 to rotate, and then drive the blades 10 on the surface of the sleeve rod 27 to rotate, thereby completing the stirring of the liquid in the fermenter 5. When the liquid in the fermenter 5 needs to be centrifuged, the user needs to pull the lifting rod 30 out of the feed port. During this process, the lifting rod 30 will lift the sleeve rod 27 through the rotating ring 28, thereby driving the sleeve rod 27 and the blades 10 away from the liquid in the fermenter 5, thereby preventing the sleeve rod 27 and the blades 10 from being inserted into the liquid, resulting in obstruction of the stratification of the liquid. To fix the lifting rod 30, it is necessary to align the insertion hole 32 at the end of the closing cover 31 with the lifting rod 30 so that the lifting rod 30 can pass through the insertion hole 32, and then screw the closing cover 31 onto the feeding pipe 7. After that, the user rotates the card plate 34 in the receiving groove 33 so that the card plate 34 rotates out of the receiving groove 33. At this time, the card plate 34 can play a limiting role to prevent the lifting rod 30 from falling back into the feeding pipe 7 and the fermenter 5.

[0059] When the fermentation tank 5 needs to be sealed and fermented, the user needs to insert the rubber plug 35 into the hole 32 to seal the hole 32, and then screw the closing cover 31 onto the feeding tube 7. At this time, the closing cover 31 can complete the sealing of the feeding tube 7. When the lifting rod 30 needs to pass through the hole 32, the user only needs to pull out the rubber plug 35 at the hole 32 to allow the lifting rod 30 to pass through the hole 32 smoothly. At the same time, the elastic rod 36 can also restrict the rubber plug 35 in the closing cover 31, thereby avoiding the loss of the rubber plug 35.

[0060] When the fermentation tank 5 needs to be centrifuged, the hydraulic cylinder 37 will drive the protective ring 38 to rise and make the roller 39 inside the protective ring 38 rest against the surface of the connecting ring 40, so that when the support arm 4 and the connecting ring 40 rotate, the connecting ring 40 provides centripetal force for each support arm 4 to avoid the support arm 4 from breaking due to centrifugal action at high speed, thereby causing the support column 3 to be thrown out and cause harm to the staff.

[0061] The above shows and describes the basic principles, main features and advantages of the present invention. It should be understood by those skilled in the art that the present invention is not limited to the above embodiments. The above embodiments and descriptions are only for explaining the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, which fall within the scope of the present invention. The scope of protection of the present invention is defined by the attached claims and their equivalents.

Claims

1. An androstenedione preparation device, characterized in that: The invention comprises a base (1); a rotating seat (2) is fixedly connected to the top surface of the base (1), and the rotating seat (2) is driven by a servo motor; a support column (3) is fixedly connected to the top surface of the rotating seat (2) at a middle position; a plurality of uniformly arranged support arms (4) are fixedly connected to the surface of the support column (3), and the number of the support arms (4) is an even number; the end of each support arm (4) away from the rotating seat (2) is rotatably connected to a fermentation tank (5); a heating component is installed on the surface of the fermentation tank (5); the heating component heats the fermentation tank (5) through an electric heating wire; a sealing cover (6) is installed at the opening of each fermentation tank (5); a feed pipe (7) and an exhaust pipe (8) are fixedly connected to the end of each sealing cover (6) away from the fermentation tank (5), and a valve is installed inside the exhaust pipe (8); a driving rod (9) is rotatably connected to the side of the sealing cover (6) close to the fermentation tank (5), and is driven by a servo motor; a blade (10) is arranged on the outer side of the driving rod (9).

2. A device for preparing androstenedione according to claim 1, characterized in that: The top surface of the rotating seat (2) is provided with a through hole (11), and the through hole (11) passes through the rotating seat (2); a hydraulic rod (12) is fixedly connected to the top surface of the base (1) at the through hole (11); the telescopic end of the hydraulic rod (12) is fixedly connected to a temporary storage tank (13); the bottom of the temporary storage tank (13) is fixedly connected to a plurality of evenly arranged support frames (14), and the support frames (14) and the support arms (4) are arranged in a one-to-one correspondence; a discharge pipe (15) is fixedly connected to one end of the support frame (14) away from the hydraulic rod (12); the position of the discharge pipe (15) corresponds to that of the feed pipe (7), and the diameter of the discharge pipe (15) is smaller than that of the feed pipe (7); a conduit (16) is fixedly connected between the discharge pipe (15) and the temporary storage tank (13), and an electric control valve is provided at the connection between the conduit (16) and the temporary storage tank (13).

3. A device for preparing androstenedione according to claim 2, characterized in that: A bellows (17) is fixedly connected to the top surface of the rotating seat (2) between two adjacent support arms (4); the diameter of the top of the bellows (17) is larger than the diameter of the discharge pipe (15) and corresponds to the position of the discharge pipe (15); a sewage outlet (18) is provided on the side surface of the rotating seat (2) at a position corresponding to the bellows (17), and the bottom of the bellows (17) is connected to the sewage outlet (18); a liquid guide ring (19) is installed on the side surface of the rotating seat (2), and one side of the liquid guide ring (19) is higher than the other side; and a notch (20) is provided at the lowest end of the liquid guide ring (19).

4. A device for preparing androstenedione according to claim 3, characterized in that: A telescopic rod (21) is fixedly connected to the bottom of the liquid guide ring (19); the telescopic rod (21) is fixed by a latch; and a baffle (22) is fixedly connected to the outer side of the liquid guide ring (19).

5. A device for preparing androstenedione according to claim 4, characterized in that: The top surface of the rotating seat (2) is rotatably connected to the outer side of the support column (3) by an internal gear (23); the inner side of the internal gear (23) is meshingly connected to a plurality of auxiliary gears (24), and the auxiliary gears (24) are rotatably connected to the top surface of the rotating seat (2); the top surface of the auxiliary gear (24) is fixedly connected to a threaded rod (25); the surface of the threaded rod (25) is threadedly connected to a movable plate (26); one end of the movable plate (26) is slidably connected to the surface of the support column (3); and the end of the movable plate (26) away from the support column (3) is fixedly connected to the top surface of the bellows (17).

6. A device for preparing androstenedione according to claim 1, characterized in that: The surface of the driving rod (9) is slidably connected to a sleeve rod (27); the blade (10) is fixedly connected to the surface of the sleeve rod (27); the end of the sleeve rod (27) close to the sealing cover (6) is rotatably connected to a rotating ring (28); one side of the rotating ring (28) is fixedly connected to a connecting plate (29); the end of the connecting plate (29) away from the rotating ring (28) is fixedly connected to a lifting rod (30), and the lifting rod (30) is located at the feed pipe (7); a closing cover (31) is threadedly connected to the feed hopper; a plug hole (32) is provided in the middle of the closing cover (31), and the plug hole (32) is compatible with the lifting rod (30); a pair of symmetrically arranged receiving grooves (33) are provided on the surface of the lifting rod (30); the end of the receiving groove (33) away from the connecting plate (29) is rotatably connected to a clamping plate (34).

7. A device for preparing androstenedione according to claim 6, characterized in that: A rubber plug (35) is provided on one side of the closing cover (31) close to the feed pipe (7) at a position corresponding to the insertion hole (32), and the diameter of the rubber plug (35) at one end close to the insertion hole (32) is smaller than that of the insertion hole (32); an elastic rod (36) is fixedly connected between the rubber plug (35) and the closing cover (31); and the diameter of the rubber plug (35) at one end close to the insertion hole (32) is smaller than the diameter of the end away from the insertion hole (32).

8. A device for preparing androstenedione according to claim 7, characterized in that: A plurality of uniformly arranged hydraulic cylinders (37) are fixedly connected around the base (1); the telescopic ends of the hydraulic cylinders (37) are commonly fixedly connected to a protective ring (38); one end of the protective ring (38) close to the support column (3) is rotatably connected to a plurality of uniformly arranged rollers (39); and a connection ring (40) is commonly installed at one end of the plurality of support arms (4) away from the support column (3).

9. A method for preparing androstenedione, the method using an androstenedione preparation device as described in any one of claims 1 to 8 above, characterized in that: The following steps are involved: S1. Take the strain, put the frozen glycerol tube of Pseudomonas alkylphenolica GR-4-4 into an ice box to thaw, inoculate it into the LB plate with an inoculation loop on an ultra-clear workbench, and culture it in a 28°C incubator for 23-24 hours. Then pick a single colony with an inoculation loop and inoculate it again into the LB plate, and culture it in a 28°C incubator for 23-24 hours. S2. Culture the strains. Use SX liquid culture medium, prepare 100 ml SX liquid culture medium in a 250 ml conical flask, and sterilize at 121°C for 30-35 min. Take the strain Pseudomonas alkylphenolica GR-4-4 plate, pick a single colony with an inoculation loop and inoculate it into the SX liquid culture medium, and then culture it in a constant temperature shaker at 28°C and 180 rpm for 15-16 h. S3, fermentation culture, 9.5-10.5L of SX liquid culture medium is introduced into the fermentation tank (5), sterilized at 121°C for 30-35min, and then the fermentation liquid is cooled to room temperature, and the cultured bacterial strain Pseudomonas alkylphenolica GR-4-4 seed liquid is aseptically poured into the fermentation tank (5), and the servo motor is started to drive the driving rod (9) to rotate, and the driving rod (9) drives the sleeve rod (27) and the blades (10) thereon to stir the liquid in the fermentation tank (5), and the fermentation is maintained at a constant temperature of 28°C for 71.5-72h; S4, extracting the transformed product, adding alkali to the fermentation liquid in the fermentation tank (5) to adjust the pH value to 11.8-12, then heating the fermentation tank (5), boiling the fermentation liquid in the fermentation tank (5) to inactivate the bacteria, then the rotating seat (2) drives the support arm (4) and the fermentation tank (5) thereon to rotate, and then the liquid in the fermentation tank (5) is separated into layers by centrifugal action, and then the supernatant in the fermentation tank (5) is collected; S5. The transformed product is initially purified, and the supernatant is concentrated to 0.9-1L in a sand bath pot. After the supernatant is cooled to room temperature, 29%-30% ethyl acetate is added to the supernatant, and the pH value is adjusted to 2-3. The temperature is controlled at 49-50°C and stirred for extraction for 5-10 minutes. The mixture is then placed in a separatory funnel and allowed to stand for separation, and the aqueous phase is collected. After repeating the extraction three times, the extracts are combined and the solvent is recovered on a rotary evaporator to 5-10ml, and then dried in a vacuum drying oven. S6. The transformed product is further purified by placing the dried product in a beaker, adding 100 times of water, adding 19% to 20% of sodium hydroxide, boiling and reacting for 29-30 minutes, cooling and adding water to 50 times, adjusting the pH value to 2-3 with hydrochloric acid at low temperature, heating and dehydrating, filtering, washing with water to a pH value of 5-6, and drying at 50°C to obtain the extracted product.

10. A method for preparing androstenedione according to claim 9, characterized in that: The fermentation tank (5) in S4 is being subjected to centrifugal separation, and it is also necessary to deflect the fermentation tank (5) so that the fermentation tank (5) is no longer perpendicular to the support arm (4), so that the stratification is more obvious. At the same time, it is also necessary to disconnect the fermentation tank (5) from the external pipeline to prevent the pipeline from being entangled when the support arm (4) drives the fermentation tank (5) to rotate.

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