Biological pharmaceutical microorganism culture method

Abstract

The invention discloses a biological pharmaceutical microorganism culture method, and relates to the technical field of microorganism gas introduction culture, and the biological pharmaceutical microorganism culture method comprises the following steps: S1, preparing a culture medium; step S2, inoculation; step S3, culturing; step S4, separation and purification; microorganisms are cultured in a constant-temperature and constant-humidity environment through the culture device, stable environmental conditions are provided for culture of the microorganisms, and growth and reproduction of the microorganisms are promoted; the culture device can accurately control the concentration of oxygen and carbon dioxide in the microorganism growth environment, oxygen needed for microorganism culture can be provided, and air pressure balance in the constant-temperature and constant-humidity box can be kept. According to the device, the rotating plate can swing in a reciprocating manner, and the culture dish in the placing groove can oscillate in a reciprocating manner, so that oxygen can be in full contact with a solution in the culture dish, the requirement of microbial cell respiration is met, and the culture efficiency and culture effect of microorganisms are improved.

Description

Technical Field

[0001] The invention relates to the technical field of microbial gas introduction culture, in particular to a biopharmaceutical microbial culture method. Background Art

[0002] In the field of biopharmaceuticals, microbial cultivation is a key technical means. Different microorganisms require different living environments. When cultivating aerobic microorganisms, it is necessary to maintain sufficient oxygen in the microbial cultivation environment, so that, as in Chinese patent CN202210554764.X, a biomedical microbial cultivation device and a cultivation method can be used. It can be seen that the existing fixed static equipment is usually used to create a growth environment for microorganisms in the culture dish, but the static cultivation equipment needs to introduce air when aerobic bacteria are cultivated, which to a certain extent will cause the accumulation of metabolic waste in the space, resulting in uneven bacterial growth inside the equipment. As for a biomedical microbial cultivation device disclosed in Chinese patent CN202310928106.7, it can be seen that the rotation of the stirring rod in the cultivation device will make the different oxygen-producing compounds fully mixed, so that the reaction rate is accelerated, and the rotation of the reaction barrel will make the compound for preparing carbon dioxide fully and evenly heated, so that the heating rate is accelerated, and then the reaction rate is accelerated, thereby greatly improving the cultivation rate of the biomedical microbial cultivation device.

[0003] Based on the search of the prior art, it can be known that when cultivating aerobic microorganisms, the cultivation device usually only focuses on a single method of filling oxygen into the microorganism cultivation environment. The microorganisms will gradually consume oxygen and produce carbon dioxide during the cultivation process. The single method of introducing oxygen into the microorganism cultivation environment used in the prior art is likely to cause an imbalance in the air pressure in the microorganism cultivation environment for a long time, resulting in excessive carbon dioxide, which affects the cultivation and reproduction rate of the microorganisms. In addition, when oxygen is introduced into the microorganism culture environment, it is not convenient to adjust the distribution range of oxygen in the microorganism culture environment, which easily leads to uneven distribution of oxygen, so that oxygen cannot fully diffuse in the microorganisms, thereby affecting the culture rate of the microorganisms. Summary of the invention

[0004] The object of the present invention is to provide a method for culturing biopharmaceutical microorganisms to solve the problems raised in the above-mentioned background technology.

[0005] To achieve the above object, the present invention provides the following technical solution: a method for culturing biopharmaceutical microorganisms, specifically comprising the following steps: Prepare the culture medium: Dissolve 1 ml of the nutrient solution in 9 ml of sterile water, shake for 5 minutes to make a 10% solution of the culture medium, inject the culture medium into the culture dish, and sterilize it with high temperature and high pressure. After sterilization, cool the culture medium to room temperature; Inoculation: Operate on a sterile operating table, use sterile tools to take a small amount of microorganisms and add them to the culture medium; Cultivation: placing the culture dish inoculated in step S2 into a culture device for cultivation, the culture device automatically maintains a constant temperature of 37°C and a constant humidity of 50%, and the culture device automatically monitors the oxygen and carbon dioxide concentrations in the microbial culture environment. When the monitored oxygen concentration is lower than a set minimum value and the carbon dioxide concentration is higher than a set maximum value, the culture device introduces oxygen into the microbial culture environment, removes carbon dioxide from the microbial culture environment, and oscillates the culture dish to disturb the gas in the microbial culture environment, thereby introducing oxygen into the culture dish; Separation and purification: After the cultivation is completed, the microbial products are separated to obtain pure microorganisms for the production of drugs.

[0006] Optionally, the culture device comprises: A constant temperature and humidity chamber, wherein a chamber door is hingedly connected to the front side of the constant temperature and humidity chamber; A culture dish placement seat, the culture dish placement seat is arranged in the constant temperature and humidity chamber, the upper end surface of the culture dish placement seat is evenly provided with placement grooves, the middle part of the lower end surface of the culture dish placement seat is fixed with a first rotating shaft, and the first rotating shaft is rotatably connected to the bottom wall of the constant temperature and humidity chamber through a bearing; A culture dish reciprocating oscillation mechanism, the culture dish reciprocating oscillation mechanism is used to drive the culture dish placement seat to reciprocate; A plurality of rotating plates, wherein the rotating plates are arranged in the constant temperature and humidity chamber, a second rotating shaft is fixedly arranged in the rotating plates, and the second rotating shaft is rotatably connected to the bottom wall of the constant temperature and humidity chamber through a bearing; a first transmission mechanism, when the culture dish reciprocating oscillation mechanism is in operation, the rotating plate is caused to reciprocate under the action of the first transmission mechanism; An oxygen introduction mechanism, an oxygen generator, a carbon dioxide derivation mechanism and a second transmission mechanism, wherein the oxygen introduction mechanism, the carbon dioxide derivation mechanism and the second transmission mechanism are all arranged in the constant temperature and humidity chamber, and the oxygen generator is arranged outside the constant temperature and humidity chamber. When the culture dish reciprocating oscillation mechanism operates, the oxygen introduction mechanism and the carbon dioxide derivation mechanism operate under the action of the second transmission mechanism, the oxygen introduction mechanism introduces the oxygen prepared by the oxygen generator into the constant temperature and humidity chamber, and the carbon dioxide derivation mechanism exports the carbon dioxide in the constant temperature and humidity chamber out of the constant temperature and humidity chamber; A control unit is used to monitor the oxygen and carbon dioxide concentrations in the constant temperature and humidity chamber and control the operation of the culture dish reciprocating oscillation mechanism and the oxygen generator.

[0007] Optionally, the culture dish reciprocating oscillation mechanism comprises: A third rotating shaft, the third rotating shaft is rotatably arranged on the inner bottom wall of the constant temperature and humidity chamber through a bearing, a motor is fixedly installed on the outer bottom of the constant temperature and humidity chamber, the output shaft of the motor is fixedly connected to the third rotating shaft, an active rotating arm is fixedly arranged on the upper end surface of the third rotating shaft, and a round pin is fixedly arranged on the side of the upper end surface of the active rotating arm away from the third rotating shaft; The driven swing arm is fixedly arranged on the outer surface of the first rotating shaft. A sliding hole is opened on the driven swing arm. The round pin extends into the sliding hole and is slidably connected with the sliding hole.

[0008] Optionally, the first transmission mechanism includes: A large gear and multiple groups of small gears, the large gear is fixed on the outer surface of the first rotating shaft, the multiple groups of small gears correspond to the second rotating shaft one by one, and the small gears are fixed on the outer surface of the second rotating shaft, and the small gears are meshed and connected with the large gear.

[0009] Optionally, the oxygen introduction mechanism includes: An oxygen introduction piston assembly comprises a first piston cylinder, a first piston plate slidably arranged in the first piston cylinder, a first piston rod fixedly arranged at the end of the first piston plate, an oxygen inlet pipe and an oxygen exhaust pipe fixedly connected to the upper wall and side wall of the first piston cylinder, a first one-way valve is installed on the oxygen inlet pipe and the oxygen exhaust pipe, the first piston cylinder is fixedly connected to the inner wall of the constant temperature and humidity chamber, and the oxygen inlet pipe extends out of the constant temperature and humidity chamber and is fixedly connected to the outlet of the oxygen generator.

[0010] Optionally, the carbon dioxide export mechanism includes: A carbon dioxide export piston assembly, the carbon dioxide export piston assembly comprising a second piston cylinder, a second piston plate slidably arranged in the second piston cylinder, a second piston rod fixedly arranged at the end of the second piston plate, a carbon dioxide inlet pipe and a carbon dioxide exhaust pipe fixedly connected to the side wall and the lower wall of the second piston cylinder, a second one-way valve being installed on the carbon dioxide inlet pipe and the carbon dioxide exhaust pipe, the second piston cylinder being fixedly connected to the inner wall of the constant temperature and humidity chamber, and the carbon dioxide exhaust pipe extending out of the constant temperature and humidity chamber.

[0011] Optionally, the oxygen exhaust pipe and the carbon dioxide inlet pipe are both facing the culture dish placement seat, and air hoods are installed at the ends of the oxygen exhaust pipe and the carbon dioxide inlet pipe. Connecting ribs are fixedly provided on the inner wall of the air hood, and a fan shaft is rotatably provided in the middle of the connecting rib through a bearing, and fan blades are evenly distributed on the outer surface of the fan shaft.

[0012] Optionally, the second transmission mechanism includes: A connecting frame, the connecting frame is fixedly arranged between the first piston rod and the second piston rod, a guide rod is fixedly arranged on the upper end of the connecting frame, a fixing block is fixedly arranged on the inner side wall of the constant temperature and humidity chamber, a guide hole for the guide rod to pass through is opened on the fixing block, a spring is connected between the fixing block and the connecting frame and located on the outer side of the guide rod, and a limit block is fixedly arranged on the upper end of the guide rod; A fourth rotating shaft, the fourth rotating shaft is rotatably connected to the inner wall of the constant temperature and humidity chamber through a bearing seat, and a cam is fixedly provided on the outer surface of the fourth rotating shaft; The fifth rotating shaft is rotatably connected to the bottom wall of the constant temperature and humidity chamber via a bearing, and the fifth rotating shaft is rotatably connected to the inner wall of the constant temperature and humidity chamber via a bearing seat, the fifth rotating shaft is transmission-connected to the fourth rotating shaft via a bevel gear transmission assembly, and the fifth rotating shaft is transmission-connected to the third rotating shaft via a pulley transmission assembly.

[0013] Optionally, the control unit includes: An oxygen concentration sensor, a carbon dioxide concentration sensor and a controller, wherein the oxygen concentration sensor and the carbon dioxide concentration sensor are fixedly installed in a constant temperature and humidity chamber, and the controller is fixedly installed outside the constant temperature and humidity chamber, and the output ends of the oxygen concentration sensor and the carbon dioxide concentration sensor are connected to the input end of the controller, and the output end of the controller is connected to the input end of the motor and the oxygen generator.

[0014] Compared with the prior art, the present invention has the following beneficial effects: 1. The present invention cultivates microorganisms in a constant temperature and humidity environment through a cultivation device, thereby providing stable environmental conditions for the cultivation of microorganisms and promoting the growth and reproduction of microorganisms; the cultivation device can accurately control the concentrations of oxygen and carbon dioxide in the microbial growth environment to meet the needs of microbial cell respiration and improve the cultivation efficiency and cultivation effect of microorganisms.

[0015] 2. The culture device used in the biopharmaceutical microorganism culture method of the present invention can automatically monitor the oxygen and carbon dioxide concentrations in the constant temperature and humidity chamber. When the oxygen concentration in the constant temperature and humidity chamber is low and the carbon dioxide concentration is high, it can automatically introduce oxygen into the constant temperature and humidity chamber and export carbon dioxide out of the constant temperature and humidity chamber, which can provide the required oxygen for the culture of microorganisms and maintain the air pressure balance in the constant temperature and humidity chamber; it can make the rotating plate swing back and forth and the culture dish in the placement tank oscillate back and forth, so that oxygen can fully contact with the solution in the culture dish, so as to meet the respiration needs of microbial cells and improve the culture efficiency and culture effect of microorganisms; First, the reciprocating oscillation mechanism of the culture dish is operated, and under the action of the second transmission mechanism, the oxygen introduction mechanism and the carbon dioxide export mechanism are driven to operate. The oxygen introduction mechanism introduces the oxygen prepared by the oxygen generator into the constant temperature and humidity chamber, and the carbon dioxide export mechanism exports the carbon dioxide in the constant temperature and humidity chamber out of the constant temperature and humidity chamber, and exchanges the air in the constant temperature and humidity chamber, which can provide the required oxygen for the cultivation of microorganisms and maintain the air pressure balance in the constant temperature and humidity chamber. Second, the culture dish reciprocating oscillation mechanism is operated to drive the culture dish placement seat and the culture dish on the upper end of the culture dish placement seat to reciprocate, so that oxygen can enter the solution in the culture dish, and the oxygen can be fully diffused in the solution to meet the breathing needs of the microbial cells; Thirdly, the reciprocating oscillation mechanism of the culture dish is operated, and under the action of the first transmission mechanism, the rotating plate is driven to swing back and forth, and the gas in the constant temperature and humidity chamber is disturbed by the reciprocating swing of the rotating plate, so that oxygen can flow in the constant temperature and humidity chamber and the oxygen can be distributed more evenly, thereby ensuring sufficient contact between the solution in the culture dish and the oxygen, and improving the efficiency of microbial culture. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 This is a flow chart of the biopharmaceutical microbial culture method of the present invention; Figure 2 It is a schematic diagram of the three-dimensional structure of the culture device of the present invention; Figure 3 It is a schematic diagram of the three-dimensional structure of the culture device of the present invention without a chamber door; Figure 4 This is a schematic diagram of the internal structure of the constant temperature and humidity chamber of the present invention; Figure 5 For the present invention Figure 4 A schematic diagram of the partially enlarged structure at center A; Figure 6 For the present invention Figure 4 Schematic diagram of the local enlarged structure at point B in the middle.

[0017] In the figure: 1, culture device; 2, constant temperature and humidity chamber; 3, chamber door; 4, culture dish placement seat; 5, placement slot; 6, first rotating shaft; 7, rotating plate; 8, second rotating shaft; 9, oxygen generator; 10, third rotating shaft; 11, motor; 12, active rotating arm; 13, round pin; 14, driven swing arm; 15, sliding hole; 16, large gear; 17, small gear; 18, oxygen introduction piston assembly; 19, oxygen inlet pipe; 20, oxygen exhaust pipe; 21, carbon dioxide extraction piston assembly; 22, carbon dioxide inlet pipe; 23, carbon dioxide exhaust pipe; 24, air hood; 25, fan blade; 26, connecting frame; 27, guide rod; 28, fixed block; 29, spring; 30, fourth rotating shaft; 31, cam; 32, fifth rotating shaft; 33, bevel gear transmission assembly; 34, pulley transmission assembly; 35, oxygen concentration sensor; 36, carbon dioxide concentration sensor; 37, controller. DETAILED DESCRIPTION

[0018] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention. Embodiment 1:

[0019] See also Figures 1 to 6 This embodiment provides a technical solution: a method for culturing biopharmaceutical microorganisms, which specifically includes the following steps: Step S1, preparing culture medium: dissolving 1 ml of nutrient solution in 9 ml of sterile water, shaking for 5 min to prepare a 10% solution culture medium, injecting the culture medium into a culture dish, and sterilizing it under high temperature and high pressure, and cooling the culture medium to room temperature after sterilization; Step S2, inoculation: operating on a sterile operating table, using sterile tools to take a small amount of microorganisms and add them into the culture medium; Step S3, culturing: placing the culture dish inoculated in step S2 into the culture device 1 for culturing, the culture device 1 automatically maintains a constant temperature of 37°C and a constant humidity of 50%, and the culture device 1 automatically monitors the oxygen and carbon dioxide concentrations in the microbial culture environment. When the monitored oxygen concentration is lower than a set minimum value and the carbon dioxide concentration is higher than a set maximum value, the culture device 1 introduces oxygen into the microbial culture environment, removes carbon dioxide from the microbial culture environment, and oscillates the culture dish to disturb the gas in the microbial culture environment, thereby introducing oxygen into the culture dish; Step S4, separation and purification: After the cultivation is completed, the microbial products are separated to obtain pure microorganisms for use in the production of drugs.

[0020] More specifically, in this embodiment: when the microorganisms are cultured in the culture device 1, a constant temperature of 37°C and a constant humidity of 50% are maintained, which provides stable environmental conditions for the culture of the microorganisms and promotes the growth and reproduction of the microorganisms; the oxygen and carbon dioxide concentrations in the culture device 1 are precisely controlled to meet the needs of microbial cell respiration and improve the culture efficiency and culture effect of the microorganisms.

[0021] Please refer to Figure 1 , Figure 2 , Figure 3 , Figure 4 , Figure 5 and Figure 6 , the culture device 1 is disclosed as follows, and the culture device 1 comprises: A constant temperature and humidity chamber 2, wherein a chamber door 3 is hingedly connected to the front side of the constant temperature and humidity chamber 2; A culture dish placement seat 4 is arranged in the constant temperature and humidity chamber 2, and a placement groove 5 is evenly opened on the upper end surface of the culture dish placement seat 4. A first rotating shaft 6 is fixedly arranged in the middle of the lower end surface of the culture dish placement seat 4, and the first rotating shaft 6 is rotatably connected to the bottom wall of the constant temperature and humidity chamber 2 through a bearing; A culture dish reciprocating oscillation mechanism, which is used to drive the culture dish placement seat 4 to reciprocate; A plurality of rotating plates 7, the rotating plates 7 are arranged in the constant temperature and humidity chamber 2, a second rotating shaft 8 is fixedly arranged in the rotating plates 7, and the second rotating shaft 8 is rotatably connected to the bottom wall of the constant temperature and humidity chamber 2 through a bearing; The first transmission mechanism, when the culture dish reciprocating oscillation mechanism is running, under the action of the first transmission mechanism, the rotating plate 7 is caused to reciprocate; The oxygen introduction mechanism, the oxygen generator 9, the carbon dioxide export mechanism and the second transmission mechanism are all arranged in the constant temperature and humidity box 2, and the oxygen generator 9 is arranged outside the constant temperature and humidity box 2. When the culture dish reciprocating oscillation mechanism is in operation, the oxygen introduction mechanism and the carbon dioxide export mechanism are operated under the action of the second transmission mechanism. The oxygen introduction mechanism introduces the oxygen prepared by the oxygen generator 9 into the constant temperature and humidity box 2, and the carbon dioxide export mechanism exports the carbon dioxide in the constant temperature and humidity box 2 out of the constant temperature and humidity box 2; The control unit is used to monitor the oxygen and carbon dioxide concentrations in the constant temperature and humidity chamber 2 and control the operation of the culture dish reciprocating oscillation mechanism and the oxygen generator 9.

[0022] More specifically, in this embodiment: the staff opens the door 3 of the constant temperature and humidity chamber 2, places the inoculated microorganism culture dish in the placement groove 5 at the upper end of the culture dish placement seat 4, and then closes the door 3, thereby placing the microorganism culture dish in the constant temperature and humidity chamber 2 for cultivation; the temperature and humidity of the constant temperature and humidity chamber 2 are set in advance according to the cultivation requirements, thereby providing a suitable temperature and humidity environment for the cultivation of microorganisms; When aerobic microorganisms are cultured, as time goes by, the microorganisms gradually consume oxygen and produce carbon dioxide. The concentrations of oxygen and carbon dioxide in the constant temperature and humidity chamber 2 are monitored by the control unit. When the monitored oxygen concentration is lower than the set minimum value and the carbon dioxide concentration is higher than the set maximum value, the control unit sends a signal to the culture dish reciprocating oscillation mechanism and the oxygen generator 9 to control the culture dish reciprocating oscillation mechanism and the oxygen generator 9 to start; The culture dish reciprocating oscillation mechanism operates to drive the culture dish placement seat 4 and the culture dish on the upper end of the culture dish placement seat 4 to reciprocate and oscillate; under the action of the first transmission mechanism, the rotating plate 7 is driven to reciprocate and swing; under the action of the second transmission mechanism, the oxygen introduction mechanism and the carbon dioxide export mechanism are driven to operate, the oxygen introduction mechanism introduces the oxygen prepared by the oxygen generator 9 into the constant temperature and humidity chamber 2, and the carbon dioxide export mechanism exports the carbon dioxide in the constant temperature and humidity chamber 2 out of the constant temperature and humidity chamber 2; Oxygen is introduced into the constant temperature and humidity chamber 2, and carbon dioxide is exported from the constant temperature and humidity chamber 2. The air in the constant temperature and humidity chamber 2 is exchanged, which can not only provide the required oxygen for the cultivation of microorganisms, but also maintain the air pressure balance in the constant temperature and humidity chamber 2. The rotating plate 7 is reciprocated to disturb the gas in the constant temperature and humidity chamber 2, so that the introduced oxygen can be more evenly distributed in the constant temperature and humidity chamber 2. At the same time, the culture dish in the placement groove 5 is reciprocated to oscillate, so that the oxygen can fully contact with the solution in the culture dish, so as to meet the respiration needs of the microbial cells and improve the culture efficiency and culture effect of the microorganisms. Embodiment 2:

[0023] Based on the above examples: Figure 3 , Figure 4 and Figure 5 The culture dish reciprocating oscillation mechanism in the first embodiment is disclosed as follows. The culture dish reciprocating oscillation mechanism comprises: A third rotating shaft 10, the third rotating shaft 10 is rotatably arranged on the inner bottom wall of the constant temperature and humidity chamber 2 through a bearing, a motor 11 is fixedly installed on the outer bottom of the constant temperature and humidity chamber 2, the output shaft of the motor 11 is fixedly connected to the third rotating shaft 10, an active rotating arm 12 is fixedly arranged on the upper end surface of the third rotating shaft 10, and a round pin 13 is fixedly arranged on the side of the upper end surface of the active rotating arm 12 away from the third rotating shaft 10; The driven swing arm 14 is fixedly arranged on the outer surface of the first rotating shaft 6 . A sliding hole 15 is opened on the driven swing arm 14 . The round pin 13 extends into the sliding hole 15 and is slidably connected with the sliding hole 15 .

[0024] More specifically, in this embodiment: the motor 11 drives the third rotating shaft 10 to rotate, so that the active rotating arm 12 and the round pin 13 rotate. When the round pin 13 is in the process of continuous rotation, the driven swing arm 14 is driven to swing back and forth, thereby driving the first rotating shaft 6 to swing back and forth at a smaller angle, and then the culture dish placement seat 4 and the culture dish placed in the placement groove 5 are caused to swing back and forth, so that oxygen can enter the solution in the culture dish, so that oxygen can be fully diffused in the solution to meet the breathing needs of the microbial cells.

[0025] Also, see Figure 3 and Figure 4 , the first transmission mechanism in the first embodiment is disclosed as follows, the first transmission mechanism comprises: The large gear 16 and multiple sets of small gears 17 , the large gear 16 is fixed on the outer surface of the first rotating shaft 6 , the multiple sets of small gears 17 correspond to the second rotating shaft 8 one by one, and the small gears 17 are fixed on the outer surface of the second rotating shaft 8 , and the small gears 17 are meshed and connected with the large gear 16 .

[0026] More specifically, in this embodiment: when the first rotating shaft 6 in the reciprocating oscillation mechanism of the culture dish performs a reciprocating swing at a small angle, it drives the large gear 16 to perform a reciprocating swing at a small angle. Since the large gear 16 is meshed and connected with the small gear 17, the small gear 17 and the second rotating shaft 8 are driven to perform a reciprocating swing, thereby driving the rotating plate 7 to perform a reciprocating swing. The reciprocating swing of the rotating plate 7 disturbs the gas in the constant temperature and humidity chamber 2, so that oxygen can flow in the constant temperature and humidity chamber 2, and the oxygen can be distributed more evenly, thereby ensuring sufficient contact between the solution in the culture dish and the oxygen, thereby improving the efficiency of microbial culture.

[0027] Also see Figure 2 , Figure 3 , Figure 4 and Figure 6 The oxygen introduction mechanism in the first embodiment is disclosed as follows. The oxygen introduction mechanism comprises: The oxygen introduction piston assembly 18 includes a first piston cylinder, a first piston plate slidably arranged in the first piston cylinder, a first piston rod fixedly arranged at the end of the first piston plate, an oxygen inlet pipe 19 and an oxygen exhaust pipe 20 fixedly connected to the upper wall and side wall of the first piston cylinder, a first one-way valve is installed on the oxygen inlet pipe 19 and the oxygen exhaust pipe 20, the first piston cylinder is fixedly connected to the inner wall of the constant temperature and humidity box 2, and the oxygen inlet pipe 19 extends out of the constant temperature and humidity box 2 and is fixedly connected to the outlet of the oxygen generator 9.

[0028] More specifically, in this embodiment: when the third rotating shaft 10 of the culture dish reciprocating oscillation mechanism rotates, under the action of the second transmission mechanism, the first piston rod and the first piston plate are driven to reciprocate in the first piston cylinder. When the first piston plate moves downward, negative pressure is generated in the first piston cylinder, and the oxygen produced by the oxygen generator 9 is sucked into the first piston cylinder through the oxygen inlet pipe 19. When the first piston plate moves upward, the oxygen sucked into the first piston cylinder is discharged from the oxygen exhaust pipe 20 to the constant temperature and humidity chamber 2. Embodiment three:

[0029] Based on the above examples: Figure 2 , Figure 3 , Figure 4 and Figure 6 The carbon dioxide export mechanism in the first embodiment is disclosed as follows. The carbon dioxide export mechanism includes: The carbon dioxide export piston assembly 21 includes a second piston cylinder, a second piston plate slidably arranged in the second piston cylinder, a second piston rod fixedly arranged at the end of the second piston plate, a carbon dioxide inlet pipe 22 and a carbon dioxide exhaust pipe 23 fixedly connected to the side wall and the lower wall of the second piston cylinder, a second one-way valve is installed on the carbon dioxide inlet pipe 22 and the carbon dioxide exhaust pipe 23, the second piston cylinder is fixedly connected to the inner wall of the constant temperature and humidity chamber 2, and the carbon dioxide exhaust pipe 23 extends out of the constant temperature and humidity chamber 2.

[0030] More specifically, in this embodiment: since the density of carbon dioxide is greater than that of oxygen, carbon dioxide will flow downwards compared to oxygen. Therefore, in this embodiment, the carbon dioxide outlet mechanism is arranged below the oxygen inlet mechanism. When the third rotating shaft 10 of the reciprocating oscillation mechanism of the culture dish rotates, under the action of the second transmission mechanism, the second piston rod and the second piston plate are driven to reciprocate in the second piston cylinder. When the second piston plate moves upward, negative pressure is generated in the second piston cylinder, and the carbon dioxide in the constant temperature and humidity chamber 2 is sucked into the second piston cylinder through the carbon dioxide inlet pipe 22. When the second piston plate moves downward, the carbon dioxide sucked into the second piston cylinder is discharged outside the constant temperature and humidity chamber 2 through the carbon dioxide exhaust pipe 23.

[0031] It is worth noting that in the present embodiment: the oxygen exhaust pipe 20 and the carbon dioxide inlet pipe 22 are both facing the culture dish placement seat 4, and gas hoods 24 are installed at the ends of the oxygen exhaust pipe 20 and the carbon dioxide inlet pipe 22. The inner wall of the gas hood 24 is fixedly provided with connecting ribs, and a fan shaft is rotatably provided in the middle of the connecting ribs through a bearing, and fan blades 25 are evenly distributed on the outer surface of the fan shaft.

[0032] More specifically, in this embodiment: when the oxygen in the first piston cylinder is discharged or the second piston cylinder inhales carbon dioxide, the fan blades 25 rotate due to the wind force, accelerating the flow of the gas, causing the gas to produce a larger vortex, thereby making the oxygen discharged into the constant temperature and humidity chamber 2 more evenly dispersed, or allowing the carbon dioxide in the constant temperature and humidity chamber 2 to enter the second piston cylinder more quickly. Embodiment 4:

[0033] Based on the above examples: Figure 4 , Figure 5 and Figure 6 , the second transmission mechanism in the first embodiment is disclosed as follows, the second transmission mechanism comprises: A connecting frame 26 is fixedly arranged between the first piston rod and the second piston rod, a guide rod 27 is fixedly arranged on the upper end of the connecting frame 26, a fixing block 28 is fixedly arranged on the inner wall of the constant temperature and humidity chamber 2, a guide hole for the guide rod 27 to pass through is opened on the fixing block 28, a spring 29 is connected between the fixing block 28 and the connecting frame 26 and located on the outer side of the guide rod 27, and a limit block is fixedly arranged on the upper end of the guide rod 27; A fourth rotating shaft 30, the fourth rotating shaft 30 is rotatably connected to the inner wall of the constant temperature and humidity chamber 2 through a bearing seat, and a cam 31 is fixedly disposed on the outer surface of the fourth rotating shaft 30; The fifth rotating shaft 32 is rotatably connected to the bottom wall of the constant temperature and humidity chamber 2 through a bearing, and the fifth rotating shaft 32 is rotatably connected to the inner wall of the constant temperature and humidity chamber 2 through a bearing seat, the fifth rotating shaft 32 is transmission-connected to the fourth rotating shaft 30 through a bevel gear transmission assembly 33, and the fifth rotating shaft 32 is transmission-connected to the third rotating shaft 10 through a pulley transmission assembly 34.

[0034] More specifically, in this embodiment: when the third rotating shaft 10 of the culture dish reciprocating oscillation mechanism rotates, the fifth rotating shaft 32 is rotated through the pulley transmission assembly 34, and the fourth rotating shaft 30 is rotated through the bevel gear transmission assembly 33. The rotation of the fourth rotating shaft 30 drives the cam 31 to rotate. During the rotation of the cam 31, the connecting frame 26 is driven to reciprocate up and down under the elastic compression or restoring force of the spring 29, thereby driving the first piston rod of the oxygen introduction mechanism and the second piston rod of the carbon dioxide export mechanism to reciprocate up and down. Embodiment five:

[0035] Based on the above examples: Figure 2 and Figure 3 , the control unit in the first embodiment is disclosed as follows, the control unit includes: The oxygen concentration sensor 35, the carbon dioxide concentration sensor 36 and the controller 37 are fixedly installed in the constant temperature and humidity chamber 2, and the controller 37 is fixedly installed outside the constant temperature and humidity chamber 2. The output ends of the oxygen concentration sensor 35 and the carbon dioxide concentration sensor 36 are connected to the input end of the controller 37, and the output end of the controller 37 is connected to the input end of the motor 11 and the oxygen generator 9.

[0036] More specifically, in this embodiment: the oxygen concentration sensor 35 and the carbon dioxide concentration sensor 36 are used to monitor the oxygen and carbon dioxide concentrations in the constant temperature and humidity chamber 2, and the monitored signals are transmitted to the controller 37. When the monitored oxygen concentration is lower than the set minimum value and the carbon dioxide concentration is higher than the set maximum value, the control unit sends a signal to the culture dish reciprocating oscillation mechanism and the oxygen generator 9 to control the culture dish reciprocating oscillation mechanism and the oxygen generator 9 to start operation.

[0037] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for culturing biopharmaceutical microorganisms, characterized in that: The specific steps include: Step S1, preparing culture medium: dissolving 1 ml of nutrient solution in 9 ml of sterile water, shaking for 5 min to prepare a 10% solution culture medium, injecting the culture medium into a culture dish, and sterilizing it under high temperature and high pressure, and cooling the culture medium to room temperature after sterilization; Step S2, inoculation: operating on a sterile operating table, using sterile tools to take a small amount of microorganisms and add them into the culture medium; Step S3, culturing: placing the culture dish inoculated in step S2 into a culture device (1) for culturing, the culture device (1) automatically maintains a constant temperature of 37°C and a constant humidity of 50%, and the culture device (1) automatically monitors the concentrations of oxygen and carbon dioxide in the microbial culture environment. When the monitored oxygen concentration is lower than a set minimum value and the carbon dioxide concentration is higher than a set maximum value, the culture device (1) introduces oxygen into the microbial culture environment and removes carbon dioxide from the microbial culture environment, and oscillates the culture dish to disturb the gas in the microbial culture environment, thereby introducing oxygen into the culture dish; Step S4, separation and purification: After the cultivation is completed, the microbial products are separated to obtain pure microorganisms for use in the production of drugs.

2. The biopharmaceutical microbial culture method according to claim 1, characterized in that: The culture device (1) comprises: A constant temperature and humidity chamber (2), wherein a chamber door (3) is hingedly connected to the front side of the constant temperature and humidity chamber (2); A culture dish placement seat (4) is arranged in the constant temperature and humidity chamber (2), the upper end surface of the culture dish placement seat (4) is evenly provided with placement grooves (5), and a first rotating shaft (6) is fixedly provided in the middle of the lower end surface of the culture dish placement seat (4), and the first rotating shaft (6) is rotatably connected to the bottom wall of the constant temperature and humidity chamber (2) via a bearing.

3. The biopharmaceutical microorganism cultivation method according to claim 2, characterized in that: The culture device (1) further comprises: A culture dish reciprocating oscillation mechanism, the culture dish reciprocating oscillation mechanism is used to drive the culture dish placement seat (4) to reciprocate; A plurality of rotating plates (7), wherein the rotating plates (7) are arranged in the constant temperature and humidity chamber (2), a second rotating shaft (8) is fixedly arranged in the rotating plates (7), and the second rotating shaft (8) is rotatably connected to the bottom wall of the constant temperature and humidity chamber (2) via a bearing; a first transmission mechanism, which, when the culture dish reciprocating oscillation mechanism is in operation, causes the rotating plate (7) to reciprocate under the action of the first transmission mechanism; An oxygen introduction mechanism, an oxygen generator (9), a carbon dioxide derivation mechanism and a second transmission mechanism, wherein the oxygen introduction mechanism, the carbon dioxide derivation mechanism and the second transmission mechanism are all arranged in the constant temperature and humidity chamber (2), and the oxygen generator (9) is arranged outside the constant temperature and humidity chamber (2). When the culture dish reciprocating oscillation mechanism is in operation, the oxygen introduction mechanism and the carbon dioxide derivation mechanism are operated under the action of the second transmission mechanism, the oxygen introduction mechanism introduces the oxygen prepared by the oxygen generator (9) into the constant temperature and humidity chamber (2), and the carbon dioxide derivation mechanism derives the carbon dioxide in the constant temperature and humidity chamber (2) out of the constant temperature and humidity chamber (2); A control unit, the control unit being used to monitor the concentrations of oxygen and carbon dioxide in the constant temperature and humidity chamber (2) and to control the operation of the culture dish reciprocating oscillation mechanism and the oxygen generator (9); The culture dish reciprocating oscillation mechanism comprises: a third rotating shaft (10), the third rotating shaft (10) being rotatably arranged on the inner bottom wall of the constant temperature and humidity chamber (2) via a bearing, a motor (11) being fixedly mounted on the outer bottom of the constant temperature and humidity chamber (2), an output shaft of the motor (11) being fixedly connected to the third rotating shaft (10), an active rotating arm (12) being fixedly arranged on the upper end surface of the third rotating shaft (10), and a round pin (13) being fixedly arranged on the side of the upper end surface of the active rotating arm (12) away from the third rotating shaft (10); A driven swing arm (14), the driven swing arm (14) being fixedly mounted on the outer surface of the first rotating shaft (6), the driven swing arm (14) being provided with a sliding hole (15), the round pin (13) extending into the sliding hole (15) and being slidably connected to the sliding hole (15); The first transmission mechanism comprises: A large gear (16) and a plurality of groups of small gears (17), wherein the large gear (16) is fixedly arranged on the outer surface of the first rotating shaft (6), the plurality of groups of small gears (17) correspond to the second rotating shaft (8) in a one-to-one manner, and the small gears (17) are fixedly arranged on the outer surface of the second rotating shaft (8), and the small gears (17) are meshingly connected with the large gear (16); The oxygen introduction mechanism comprises: An oxygen introduction piston assembly (18) comprises a first piston cylinder, a first piston plate slidably arranged in the first piston cylinder, a first piston rod fixedly arranged at the end of the first piston plate, an oxygen inlet pipe (19) and an oxygen exhaust pipe (20) fixedly connected to the upper wall and side wall of the first piston cylinder, a first non-return valve being installed on both the oxygen inlet pipe (19) and the oxygen exhaust pipe (20), the first piston cylinder being fixedly connected to the inner wall of the constant temperature and humidity chamber (2), the oxygen inlet pipe (19) extending out of the constant temperature and humidity chamber (2) and fixedly connected to the outlet of the oxygen generator (9).

4. The biopharmaceutical microorganism cultivation method according to claim 3, characterized in that: The carbon dioxide export mechanism comprises: A carbon dioxide export piston assembly (21), the carbon dioxide export piston assembly (21) comprising a second piston cylinder, a second piston plate slidably disposed in the second piston cylinder, a second piston rod fixedly disposed at the end of the second piston plate, a carbon dioxide intake pipe (22) and a carbon dioxide exhaust pipe (23) fixedly connected to the side wall and the lower wall of the second piston cylinder, and a second one-way valve is installed on each of the carbon dioxide intake pipe (22) and the carbon dioxide exhaust pipe (23).

5. The biopharmaceutical microorganism cultivation method according to claim 4, characterized in that: The second piston cylinder is fixedly connected to the inner wall of the constant temperature and humidity chamber (2), and the carbon dioxide exhaust pipe (23) extends out of the constant temperature and humidity chamber (2).

6. The biopharmaceutical microbial culture method according to claim 5, characterized in that: The oxygen exhaust pipe (20) and the carbon dioxide intake pipe (22) are both directed toward the culture dish placement seat (4); gas hoods (24) are installed at the ends of the oxygen exhaust pipe (20) and the carbon dioxide intake pipe (22); a connecting rib is fixedly provided on the inner wall of the gas hood (24); a fan shaft is rotatably provided in the middle of the connecting rib via a bearing; and fan blades (25) are evenly distributed on the outer surface of the fan shaft; The second transmission mechanism comprises: A connecting frame (26), the connecting frame (26) being fixedly arranged between the first piston rod and the second piston rod, a guide rod (27) being fixedly arranged on the upper end of the connecting frame (26), a fixing block (28) being fixedly arranged on the inner side wall of the constant temperature and humidity chamber (2), a guide hole for the guide rod (27) to pass through being opened on the fixing block (28), a spring (29) being connected between the fixing block (28) and the connecting frame (26) and located on the outer side of the guide rod (27), and a limit block being fixedly arranged on the upper end of the guide rod (27); A fourth rotating shaft (30), the fourth rotating shaft (30) being rotatably connected to the inner wall of the constant temperature and humidity chamber (2) via a bearing seat, and a cam (31) being fixedly provided on the outer surface of the fourth rotating shaft (30).

7. The biopharmaceutical microorganism cultivation method according to claim 6, characterized in that: The second transmission mechanism further comprises: A fifth rotating shaft (32), the fifth rotating shaft (32) is rotatably connected to the bottom wall of the constant temperature and humidity chamber (2) via a bearing, and the fifth rotating shaft (32) is rotatably connected to the inner wall of the constant temperature and humidity chamber (2) via a bearing seat, the fifth rotating shaft (32) is transmission-connected to the fourth rotating shaft (30) via a bevel gear transmission assembly (33), and the fifth rotating shaft (32) is transmission-connected to the third rotating shaft (10) via a pulley transmission assembly (34).

8. The biopharmaceutical microorganism cultivation method according to claim 7, characterized in that: The control unit comprises: An oxygen concentration sensor (35), a carbon dioxide concentration sensor (36) and a controller (37), wherein the oxygen concentration sensor (35) and the carbon dioxide concentration sensor (36) are both fixedly installed in a constant temperature and humidity chamber (2), and the controller (37) is fixedly installed outside the constant temperature and humidity chamber (2).

9. The biopharmaceutical microbial culture method according to claim 8, characterized in that: The output ends of the oxygen concentration sensor (35) and the carbon dioxide concentration sensor (36) are connected to the input end of the controller (37), and the output end of the controller (37) is connected to the input end of the motor (11) and the oxygen generator (9).

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