Bioreactor device

Abstract

The utility model relates to the technical field of biological product production, in particular to a bioreactor device. Comprising a three-necked flask, the lower end of the three-necked flask is located in a constant-temperature water bath kettle, a stirring device is installed in a middle port of the three-necked flask, a pH meter probe is installed in a side port in one side of the three-necked flask, a liquid adding pipe is installed in a side port in the other side of the three-necked flask, and the pH meter probe is electrically connected with a control panel. The control panel is electrically connected with the pH meter-peristaltic pump linkage controller, the control panel is electrically connected with the peristaltic pump, the liquid inlet end of the peristaltic pump is communicated with the liquid supplementing bottle, alkali liquor or acid liquor is stored in the liquid supplementing bottle, the liquid outlet end of the peristaltic pump is communicated with the liquid adding pipe, a support is fixed in the constant-temperature water bath kettle, and the three-neck flask is stabilized on the support. The bracket is arranged in the constant-temperature water bath kettle and can stabilize the three-neck flask, so that the stability of the three-neck flask in the water bath kettle can be improved.

Description

Technical Field

[0001] The utility model relates to the technical field of biological product production, and particularly relates to a bioreactor device. Background Art

[0002] The research and development background of the bioreactor device is based on the need for the control and optimization of biological processes, and the application scenarios cover multiple fields such as biopharmaceuticals, biovaccine production, new drug research and development, food industry, and chemical industry. During the biological conversion process in the bioreactor device, a constant pH value and a constant temperature need to be ensured.

[0003] A patent document with the authorization announcement number CN112280758B discloses a steroid 5β-reductase variant and its uses. In Example 2 of the specification of this patent document, it is recorded that the CmP5βR-LY enzyme uses NADH as a coenzyme to prepare 5β-pregnane-3,20-dione.

[0004] The following steps are used to prepare 5β-pregnane-3,20-dione:

[0005] (1) Transform the pET26b(+) vector recombinant expression plasmid containing the CmP5βR-LY coding gene into the competent cells of the highly expressing strain E. coli T7 to obtain a recombinant bacterium.

[0006] (2) Inoculate a single colony of the recombinant bacterium into an LBK (LB medium containing 50 μg / mL kanamycin sulfate) medium and culture it overnight at 37°C.

[0007] (3) Transfer 50 μL of the bacterial liquid to 5 mL of fresh LBK medium and culture it at 37°C for 2 h until the OD(600 nm) value is approximately equal to 0.6. Subsequently, add IPTG to a final concentration of 1 mM and culture it overnight at 22°C for 20 h.

[0008] (4) Centrifuge to collect the induced bacteria and wash them twice with Tris-HCl (pH 5.5) buffer. Lyse the cells by low-temperature ultrasonic waves to obtain a lysate.

[0009] (5) Take 300 μL of the above lysate, add coenzyme NADH to a final concentration of 3 mg / mL, and add the substrate progesterone to a final concentration of 0.2 mg / mL. After the entire reaction system reacts at 40°C for 4 h, add 2 volumes of ethyl acetate to terminate the reaction process.

[0010] (6) Vortex and extract the above reaction product, take the supernatant to evaporate the solvent, and redissolve it to perform TLC detection, GC detection, MS detection, and NMR detection for qualitative and quantitative analysis of the product.

[0011] In step (5) of Example 2 of this patent document, a bioreactor device is required to ensure that the reaction solution maintains constant pH and temperature conditions. However, the scale of the biological reaction system involved in this step is too small, restricting our further exploration in terms of scaling up the reaction conditions. Additionally, a previous patent application of the applicant, CN201710291852, which relates to an engineered bacterium and its use for preparing testosterone, also mentions a similar reaction process, but still uses a small-scale system and also faces technical challenges when scaling up the reaction system.

[0012] Regarding temperature control, the common practice is to place the bioreactor in a constant temperature water bath to maintain its temperature stability. However, the low-cost bioreactor devices currently on the market do not have the function of automatically adjusting the pH value of the reaction solution in the bioreactor, which may lead to fluctuations in the pH value during the reaction process, thereby having an adverse impact on the conversion rate of the reaction product. Therefore, in future research and practice, special attention needs to be paid to and the equipment performance in this aspect needs to be optimized to improve the overall reaction efficiency and product quality. Summary of the Utility Model

[0013] The technical problem to be solved by the present utility model is to provide a bioreactor device with low cost and excellent performance, which can automatically adjust the pH value of the reaction solution in the bioreactor while maintaining a constant temperature, and at the same time ensure the stability of the reaction solution container of the bioreactor device in a constant temperature water bath.

[0014] To achieve the above object, the technical solution provided by the present utility model is:

[0015] A bioreactor device includes a three-necked flask. The lower end of the three-necked flask is located in a constant temperature water bath. A stirring device is installed in the middle port of the three-necked flask. A pH probe is installed in the side port on one side of the three-necked flask. A liquid adding tube is installed in the side port on the other side of the three-necked flask. The pH probe is electrically connected to a control panel. The control panel is electrically connected to a pH meter - peristaltic pump linkage controller. The control panel is electrically connected to a peristaltic pump. The liquid inlet end of the peristaltic pump is communicated with a replenishing bottle, and an alkali solution or an acid solution is stored in the replenishing bottle. The liquid outlet end of the peristaltic pump is communicated with the liquid adding tube. A bracket is fixed in the constant temperature water bath, and the three-necked flask is stabilized on the bracket.

[0016] Specifically, a display screen, a flow rate adjustment button, and an acid / alkali adjustment button are provided on the control panel.

[0017] Specifically, the stirring device is a stirring paddle, and the stirring paddle is driven by a motor. The motor is electrically connected to the pH meter - peristaltic pump linkage controller.

[0018] Specifically, the bracket includes a support plate in an inverted U shape. The vertical part of the support plate is fixed in a constant temperature water bath. A round hole is formed in the middle of the flat part of the support plate. The spherical part at the lower side of the three-necked flask is seated in the round hole. Vertical plates are fixed on the support plates on both sides of the three-necked flask. The two vertical plates correspond to the two side ports of the three-necked flask one by one. A clamping plate is elastically rotatably connected to the upper end of the vertical plate. A clamping groove is formed in the clamping plate. The side port of the three-necked flask is clamped in the clamping groove of the clamping plate on one side. An ear plate is fixed on the clamping plate. A vertical optical rod is rotatably connected to the ear plate. The optical rod passes through a long hole in the flat part of the support plate. A pressing rod is fixed at the lower end of the optical rod. The pressing rod is located inside the support plate. The spherical part of the three-necked flask tightly abuts against the pressing rod.

[0019] Specifically, the diameter of the round hole is smaller than the diameter of the spherical part at the lower side of the three-necked flask.

[0020] Specifically, the clamping groove is a V-shaped groove.

[0021] Specifically, the vertical plate and the clamping plate are rotatably connected through a rotating shaft. A torsion spring is sleeved on the rotating shaft. One end of the torsion spring is fixedly connected to the vertical plate, and the other end of the torsion spring is fixedly connected to the clamping plate. When the pressing rod and the optical rod are pushed downward under the gravity of the three-necked flask, the optical rod can make the clamping plate rotate toward the three-necked flask by overcoming the elastic force of the torsion spring through the ear plate.

[0022] Compared with the prior art, the beneficial effects of the present utility model are as follows:

[0023] 1. The pH meter probe can monitor the pH value of the reaction solution in the three-necked flask in real time. When the pH value of the reaction solution in the three-necked flask changes, the pH meter probe sends a signal to the pH meter - peristaltic pump linkage controller. Then, the pH meter - peristaltic pump linkage controller starts the peristaltic pump to add acid solution or alkali solution in the liquid replenishing bottle into the three-necked flask to adjust the pH value of the reaction solution in the three-necked flask, which can effectively ensure the constancy of the pH value in the three-necked flask, improve the conversion rate of the reaction product, and realize the automatic adjustment of the pH value of the reaction solution in the three-necked flask.

[0024] 2. When the pH value of the reaction solution in the three-necked flask is automatically adjusted, the pH meter - peristaltic pump linkage controller starts the motor, and then the stirring paddle stirs the reaction solution in the three-necked flask, which can make the acid solution or alkali solution entering the three-necked flask be quickly mixed with the reaction solution, improve the accuracy of the pH meter probe for real-time monitoring of the pH value of the reactants in the three-necked flask, and avoid adding too much acid solution or alkali solution into the three-necked flask.

[0025] 3. The control panel is provided with a flow rate adjustment button. The output flow rate of the peristaltic pump can be adjusted through the flow rate adjustment button. When the amount of the reaction solution in the three-necked flask is large, the output flow rate after the peristaltic pump is started can be increased, and the amount of the acid solution or the alkali solution entering the three-necked flask per unit time can be increased, so that the pH value of the reaction solution in the three-necked flask can still be quickly and automatically adjusted when the amount of the reaction solution in the three-necked flask is large.

[0026] 4. The control panel is provided with an acid / alkali adjustment button. The pH threshold value when the pH meter - peristaltic pump linkage controller starts the peristaltic pump can be set through the acid / alkali adjustment button, so that the bioreactor device has a wide applicable range.

[0027] 5. By arranging a bracket in the constant temperature water bath, the bracket can stabilize the three-necked flask and improve the stability of the three-necked flask in the water bath. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 It is a schematic diagram of the bioreactor device of the present utility model.

[0029] Figure 2 is Figure 1 a schematic diagram of the bracket in

[0030] The names of the components in the drawings are: 1. pH meter - peristaltic pump linkage controller, 2. Control panel, 3. Peristaltic pump, 4. Liquid replenishing bottle, 5. Constant temperature water bath, 6. Three-necked flask, 7. pH meter probe, 8. Liquid adding tube, 9. Stirring device, 10. Bracket, 101. Support plate, 102. Vertical plate, 103. Clamping plate, 104. Card slot, 105. Ear plate, 106. Smooth rod, 107. Pressing rod, 108. Round hole, 109. Long hole. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0031] Next, the technical solutions in the embodiments of the present utility model will be clearly and completely described in conjunction with the drawings in the embodiments of the present utility model. Obviously, the described embodiments are only a part of the embodiments of the present utility model, rather than all the embodiments.

[0032] Example 1

[0033] Referring to Figure 1 as shown, a bioreactor device includes a three-necked flask 6. The lower end of the three-necked flask 6 is located in a constant temperature water bath 5, and a stirring device 9 is installed in the middle port of the three-necked flask 6.

[0034] The stirring device 9 is a stirring paddle, and the stirring paddle is driven by a motor.

[0035] A pH meter probe 7 is installed in the side port on one side of the three-necked flask 6. A liquid adding tube 8 is installed in the side port on the other side of the three-necked flask 6.

[0036] The pH meter probe 7 is electrically connected to the control panel 2, the control panel 2 is electrically connected to the pH meter - peristaltic pump linkage controller 1, and the control panel 2 is electrically connected to the peristaltic pump 3. The motor is electrically connected to the pH meter - peristaltic pump linkage controller 1.

[0037] The liquid inlet end of the peristaltic pump 3 is communicated with the replenishing bottle 4, and the replenishing bottle 4 stores alkaline liquid or acidic liquid. The liquid outlet end of the peristaltic pump 3 is communicated with the liquid adding tube 8.

[0038] The pH meter probe 7 can monitor the pH value of the reaction liquid in the three - necked flask 6 in real time. When the pH value of the reaction liquid in the three - necked flask 6 changes, the pH meter probe 7 sends a signal to the pH meter - peristaltic pump linkage controller 1, and then the pH meter - peristaltic pump linkage controller 1 starts the peristaltic pump 3 to add the acidic liquid or alkaline liquid in the replenishing bottle 4 into the three - necked flask 6 to adjust the pH value of the reaction liquid in the three - necked flask 6, which can effectively ensure the constancy of the pH value in the three - necked flask 6, improve the conversion rate of the reaction product, and realize the automatic adjustment of the pH value of the reaction liquid in the three - necked flask 6.

[0039] A display screen, a flow rate adjustment button, and an acid / alkali adjustment button are arranged on the control panel 2. The pH value of the reaction liquid in the three - necked flask 6 can be displayed in real time on the display screen of the control panel 2.

[0040] The output flow rate of the peristaltic pump 3 can be adjusted through the flow rate adjustment button. When the amount of the reaction liquid in the three - necked flask 6 is large, the output flow rate after the peristaltic pump 3 is started can be increased to increase the amount of the acidic liquid or alkaline liquid entering the three - necked flask 6 per unit time, and the rapid automatic adjustment of the pH value of the reaction liquid in the three - necked flask 6 can still be realized when the amount of the reaction liquid in the three - necked flask 6 is large.

[0041] The pH threshold value when the pH meter - peristaltic pump linkage controller 1 starts the peristaltic pump 3 can be set through the acid / alkali adjustment button, so that the biological reactor device has a wide applicable range.

[0042] Example 2

[0043] On the basis of Example 1, with reference to Figure 1 and Figure 2 as shown, a bracket 10 is fixed in the constant - temperature water bath 5, and the three - necked flask 6 is stably placed on the bracket 10.

[0044] The bracket 10 includes a U - shaped support plate 101, and the vertical part of the support plate 101 is fixed in the constant - temperature water bath 5. A round hole 108 is opened in the middle of the flat part of the support plate 101. Specifically, the diameter of the round hole 108 is smaller than the diameter of the lower spherical part of the three - necked flask 6. The lower spherical part of the three - necked flask 6 is seated in the round hole 108.

[0045] On both sides of the three-necked flask 6, there are fixed vertical plates 102 on the support plates 101. The two vertical plates 102 correspond to the two side ports of the three-necked flask 6 one by one. The upper end of the vertical plate 102 is elastically rotatably connected to a clamping plate 103, and a clamping groove 104 is formed on the clamping plate 103. Specifically, the clamping groove 104 is a V-shaped groove. The side port of the three-necked flask 6 is clamped in the clamping groove 104 of the clamping plate 103 on one side thereof.

[0046] The vertical plate 102 and the clamping plate 103 are rotatably connected through a rotating shaft, and a torsion spring is sleeved on the rotating shaft. One end of the torsion spring is fixedly connected to the vertical plate 102, and the other end of the torsion spring is fixedly connected to the clamping plate 103.

[0047] A lug plate 105 is fixed on the clamping plate 103, and a vertical polished rod 106 is rotatably connected to the lug plate 105. The polished rod 106 passes through a long hole 109 on the straight part of the support plate 101. A pressing rod 107 is fixed at the lower end of the polished rod 106. The pressing rod 107 is located inside the support plate 101, and the spherical part of the three-necked flask 6 abuts tightly against the pressing rod 107.

[0048] When the three-necked flask 6 presses the pressing rod 107 and the polished rod 106 to move downward under the action of gravity, the polished rod 106 can make the clamping plate 103 rotate towards the three-necked flask 6 against the elastic force of the torsion spring through the lug plate 105.

[0049] When the three-necked flask 6 is placed into the constant temperature water bath 5, the three-necked flask 6 moves downward between the two clamping plates 103 and the two vertical plates 102, and the two vertical plates 102 correspond to the two side ports of the three-necked flask 6 one by one. During the process of the spherical part at the lower side of the three-necked flask 6 sitting on the round hole 108, when the spherical part at the lower side of the three-necked flask 6 contacts the pressing rod 107, under the action of the gravity of the three-necked flask 6, the pressing rod 107 and the polished rod 106 can be pressed to move downward, and the polished rod 106 can make the clamping plate 103 rotate towards the three-necked flask 6 against the elastic force of the torsion spring through the lug plate 105. When the spherical part at the lower side of the three-necked flask 6 sits in the round hole 108, the spherical part of the three-necked flask 6 abuts tightly against the pressing rod 107, and the side ports on both sides of the three-necked flask 6 are respectively clamped in the clamping grooves 104 of the clamping plates 103 on both sides. The bracket 10 can stabilize the three-necked flask 6 in the constant temperature water bath 5, and can improve the stability of the three-necked flask 6 in the water bath.

[0050] The above are only the preferred embodiments of the present utility model, and are not intended to limit the present utility model. Any modifications, equivalent replacements, and improvements made within the spirit and principle of the present utility model shall be included within the protection scope of the present utility model.

Claims

1. A bioreactor device, comprising a three-necked flask (6), the lower end of the three-necked flask (6) being located in a thermostatic water bath (5), and a stirring device (9) being installed in the middle port of the three-necked flask (6), characterized in that, A pH probe (7) is installed inside the side port on one side of the three-necked flask (6), a liquid adding tube (8) is installed inside the side port on the other side of the three-necked flask (6), the pH probe (7) is electrically connected to the control panel (2), the control panel (2) is electrically connected to the pH-meter peristaltic pump linkage controller (1), the control panel (2) is electrically connected to the peristaltic pump (3), the liquid inlet end of the peristaltic pump (3) is communicated with the replenishing bottle (4), an alkali solution or an acid solution is stored in the replenishing bottle (4), the liquid outlet end of the peristaltic pump (3) is communicated with the liquid adding tube (8), a bracket (10) is fixed in the constant temperature water bath (5), and the three-necked flask (6) is stably placed on the bracket (10).

2. The bioreactor device according to claim 1, characterized in that, A display screen, a flow rate adjusting button and an acid / alkali adjusting button are arranged on the control panel (2).

3. The bioreactor device according to claim 1, characterized in that, The stirring device (9) is a stirring paddle, the stirring paddle is driven by a motor, and the motor is electrically connected to the pH-meter peristaltic pump linkage controller (1).

4. The bioreactor device according to claim 1, characterized in that, The bracket (10) includes a U-shaped inverted support plate (101), the vertical part of the support plate (101) is fixed in the constant temperature water bath (5), a round hole (108) is formed in the middle of the flat part of the support plate (101), the spherical part on the lower side of the three-necked flask (6) is seated in the round hole (108), vertical plates (102) are fixed on the support plates (101) on both sides of the three-necked flask (6), the two vertical plates (102) correspond to the side ports of the three-necked flask (6) one by one, a clamping plate (103) is elastically rotatably connected to the upper end of the vertical plate (102), a clamping groove (104) is formed in the clamping plate (103), the side port of the three-necked flask (6) is clamped in the clamping groove (104) of the clamping plate (103) on one side, an ear plate (105) is fixed on the clamping plate (103), a vertical polished rod (106) is rotatably connected to the ear plate (105), the polished rod (106) penetrates through a long hole (109) in the flat part of the support plate (101), a pressing rod (107) is fixed at the lower end of the polished rod (106), the pressing rod (107) is located inside the support plate (101), and the spherical part of the three-necked flask (6) tightly abuts against the pressing rod (107).

5. The bioreactor device according to claim 4, characterized in that, The diameter of the round hole (108) is smaller than the diameter of the spherical part on the lower side of the three-necked flask (6).

6. The bioreactor device according to claim 4, wherein The clamping groove (104) is a V-shaped groove.

7. The bioreactor device according to claim 4, characterized in that, The vertical plate (102) and the clamping plate (103) are rotatably connected through a rotating shaft, a torsion spring is sleeved on the rotating shaft, one end of the torsion spring is fixedly connected to the vertical plate (102), the other end of the torsion spring is fixedly connected to the clamping plate (103), when the pressing rod (107) and the polished rod (106) are pushed downward under the gravity of the three-necked flask (6), the polished rod (106) can enable the clamping plate (103) to rotate towards the three-necked flask (6) against the elastic force of the torsion spring through the ear plate (105).

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