CN108048323B - Alkali supplementing equipment and method for bioreactor - Google Patents
Alkali supplementing equipment and method for bioreactor Download PDFInfo
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- CN108048323B CN108048323B CN201810073816.5A CN201810073816A CN108048323B CN 108048323 B CN108048323 B CN 108048323B CN 201810073816 A CN201810073816 A CN 201810073816A CN 108048323 B CN108048323 B CN 108048323B
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- 239000003513 alkali Substances 0.000 title claims abstract description 90
- 230000001502 supplementing effect Effects 0.000 title claims abstract description 27
- 238000000034 method Methods 0.000 title claims abstract description 16
- 238000003756 stirring Methods 0.000 claims abstract description 105
- 238000005507 spraying Methods 0.000 claims abstract description 22
- 239000007788 liquid Substances 0.000 claims description 33
- 238000009423 ventilation Methods 0.000 claims description 7
- 230000009469 supplementation Effects 0.000 claims 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 abstract description 17
- 229910052760 oxygen Inorganic materials 0.000 abstract description 17
- 239000001301 oxygen Substances 0.000 abstract description 17
- 238000004113 cell culture Methods 0.000 abstract description 9
- 230000006378 damage Effects 0.000 abstract description 4
- 230000030833 cell death Effects 0.000 abstract description 3
- 210000004102 animal cell Anatomy 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 14
- 210000004027 cell Anatomy 0.000 description 12
- 230000000052 comparative effect Effects 0.000 description 8
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 8
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 6
- 239000004310 lactic acid Substances 0.000 description 4
- 235000014655 lactic acid Nutrition 0.000 description 4
- 239000001569 carbon dioxide Substances 0.000 description 3
- 229910002092 carbon dioxide Inorganic materials 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 239000002699 waste material Substances 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 238000005276 aerator Methods 0.000 description 2
- 230000003833 cell viability Effects 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 230000002503 metabolic effect Effects 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 230000005779 cell damage Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 208000037887 cell injury Diseases 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000013065 commercial product Substances 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000036284 oxygen consumption Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/08—Bioreactors or fermenters specially adapted for specific uses for producing artificial tissue or for ex-vivo cultivation of tissue
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
- C12M27/02—Stirrer or mobile mixing elements
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/06—Nozzles; Sprayers; Spargers; Diffusers
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Biomedical Technology (AREA)
- Genetics & Genomics (AREA)
- Sustainable Development (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention relates to the technical field of animal cell culture, in particular to a device and a method for supplementing alkali for a bioreactor; the apparatus includes a tank; a stirring device is arranged in the tank body; an alkali liquor spraying head is arranged above the tank wall of the tank body, and the alkali liquor spraying head is connected with an alkali storage container through a valve. According to the invention, by adopting a pulse alkali liquor supplementing mode and matching with a stirring device and a baffle which are specially arranged, turbulent flow generated during stirring can be effectively reduced, alkali liquor can be rapidly mixed uniformly, and thus cell death caused by mechanical damage and overhigh local alkali liquor concentration is reduced. In addition, unexpectedly, the unique stirring paddle rotating mode can also prolong the residence time of the oxygen, thereby effectively reducing the consumption of the oxygen.
Description
Technical Field
The invention relates to the technical field of animal cell culture, in particular to a device and a method for supplementing alkali for a bioreactor.
Background
Cell culture starts from the beginning of the century, and the scale of the cell culture starts to expand in 1962, so that the development has become a widely adopted technology in biological and medical research and application, and the production of enzymes, growth factors, vaccines, monoclonal antibodies and the like with important medical values by using the cell culture has become an important part of the medical biotechnology industry.
The accumulation of metabolic waste products in the cellular environment, including carbon dioxide, lactic acid and ammonia, is a major factor in inhibiting cell growth. Carbon dioxide and lactic acid are the most important acid metabolic wastes in the culture medium, and with the increase of the cell culture scale and density, the carbon dioxide and the lactic acid are gradually accumulated in the cell culture medium, so that the inhibition and even the toxic action on the normal physiological functions of cells are generated.
In the prior art, the basic substances (such as NaHCO) are added to the cell culture solution 3 Solution) to neutralize the lactic acid, and fed into the tank along the wall of the tank by a non-pressure drop method (as shown in fig. 1). The feeding way is slow, and the cells around the alkali liquor are lost and die due to the fact that the cells can not be dissolved quickly after being dripped into the tank, and waste of the alkali liquor is large.
In view of this, the present invention has been made.
Disclosure of Invention
The invention relates to a bioreactor alkali supplementing device, which comprises a tank body;
a stirring device is arranged in the tank body;
an alkali liquor spraying head is arranged above the tank wall of the tank body, and the alkali liquor spraying head is connected with an alkali storage container through a valve.
The device provided by the invention can ensure that the alkali liquor is stirred at the highest speed, thereby reducing the death quantity of cells.
The invention also relates to a method for supplementing alkali in the bioreactor, when the equipment is used for supplementing alkali, the valve is intermittently opened, so that the alkali liquid injection head can be used for injecting alkali liquid in a pulse mode, and meanwhile, the stirring device is opened, so that the supplemented alkali liquid is completely mixed rapidly.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described, and it is obvious that the drawings in the description below are some embodiments of the present invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 is a schematic diagram of a basic filling device in the prior art;
FIG. 2 is a schematic diagram of an alkali-supplementing device according to the present invention;
FIG. 3 is a schematic diagram of an alkali-compensating apparatus according to an embodiment of the present invention;
FIG. 4 is a schematic top view of an alkali replenishment apparatus used in one embodiment of the present invention.
Reference numerals:
a tank body A;
microbubble ventilation device 1
A stirring device 2; a stirring shaft 201; a stirring paddle 202;
a power device 3;
a baffle 4;
an alkali liquor spraying head 5; and a valve 501.
Detailed Description
The technical solution of the present invention will be clearly and completely described below with reference to the accompanying drawings and detailed description, but it will be understood by those skilled in the art that the examples described below are some, but not all, examples of the present invention, and are intended to be illustrative of the present invention only and should not be construed as limiting the scope of the present invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
In the description of the present invention, it should be noted that the directions or positional relationships indicated by the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc. are based on the directions or positional relationships shown in the drawings, are merely for convenience of describing the present invention and simplifying the description, and do not indicate or imply that the devices or elements referred to must have a specific orientation, be configured and operated in a specific orientation, and thus should not be construed as limiting the present invention. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
In the description of the present invention, it should be noted that, unless explicitly specified and limited otherwise, the terms "mounted," "connected," and "connected" are to be construed broadly, and may be either fixedly connected, detachably connected, or integrally connected, for example; can be mechanically or electrically connected; can be directly connected or indirectly connected through an intermediate medium, and can be communication between two elements. The specific meaning of the above terms in the present invention will be understood in specific cases by those of ordinary skill in the art.
The invention relates to an alkali supplementing device of a bioreactor, which is shown in figure 3, and comprises a tank body A;
a stirring device 2 is arranged in the tank A;
an alkali liquor spraying head 5 is arranged above the tank wall of the tank body A, and the alkali liquor spraying head 5 is connected with an alkali storage container through a valve 501.
The principle of the device is shown in figure 2, the alkali liquor spraying head 5 sprays alkali liquor into the culture medium of the tank body in a pulse mode, and the culture medium is rapidly stirred and uniformly mixed in the stirring device 2, so that cell damage caused by local overhigh pH can be avoided.
Preferably, a pH meter is further arranged in the alkali supplementing equipment of the bioreactor.
Preferably, in the alkali supplementing device of the bioreactor, the aperture of the inner hole at the front end of the alkali liquor spraying head 5 is 1 mm-3 mm; more preferably 2mm.
Preferably, the alkali supplementing device of the bioreactor comprises a stirring shaft 201 and a stirring paddle 202;
the stirring shaft 201 is perpendicular to the horizontal plane and fixed at the bottom of the tank, the stirring paddle 202 is arranged on the stirring shaft 201, and the bottom of the stirring shaft 201 is connected with the power device 3;
preferably, the apparatus further comprises a micro-bubble aerator 1, the micro-bubble aerator 1 being arranged directly below the stirring paddle 202.
Microbubble ventilation device in the present inventionThe device 1 can be a commercial product and can supply oxygen/CO to cells 2 And (3) obtaining the product.
Preferably, in the above-mentioned alkali supplementing device for bioreactor, the stirring paddles 202 are three, and the stirring shaft is provided with a first stirring paddle, a second stirring paddle and a third stirring paddle from top to bottom in sequence; the second stirring paddle stirs the liquid to move from top to bottom; the first stirring paddles and the third stirring paddles stir the liquid to move from bottom to top;
through the arrangement mode, oxygen released by the micro-bubble ventilation device 1 can be trapped between the first stirring paddle and the third stirring paddle (the spiral liquid flow is formed between the three stirring paddles), so that the residence time of the oxygen micro-bubbles in the culture solution is prolonged, and the utilization rate of the oxygen is increased.
Preferably, the stirring paddle 202 is a turbine type stirring paddle or a propulsion type stirring paddle;
more preferably, the paddles 202 are four-bladed propeller paddles.
Preferably, the blade inclination angle of the stirring paddle is 40-60 degrees; more preferably 50 °; since each paddle causes the direction of movement of the liquid stream to be different, it is preferable that the deflection angle of the second paddle is mirror symmetrical to the deflection angles of the first and third paddles.
Preferably, the ratio of the diameter of the stirring paddle to the diameter of the tank is 0.2-0.7, more preferably 0.3-0.5.
Preferably, the rotation speed of the stirring paddle is 30-70 rpm.
Preferably, in the alkali supplementing device of the bioreactor, a baffle plate 4 is further arranged inside the tank wall of the tank body, and the baffle plate 4 is perpendicular to the horizontal plane;
preferably, 3 to 5 baffles 4, more preferably 4 baffles are uniformly distributed on the tank wall of the tank body;
preferably, as shown in fig. 4, the baffles 4 are deviated from the warp line at which the connection point of the baffles and the tank A is positioned by 30-40 degrees, and the deflection directions of the baffles are consistent;
preferably, the width of the baffle plate is 1/13-1/10 of the tank diameter.
More preferably, the deflection direction of each baffle plate is consistent with the rotation direction of the stirring paddle; for example, the rotation direction of the stirring paddle is anticlockwise, and the deflection direction of the baffle plate 4 is shown in fig. 4, and the flow of water is deflected at an obtuse angle along the anticlockwise direction.
Because the second stirring paddle and the first stirring paddle and the third stirring paddle stir the liquid in opposite directions, the flowing direction of the liquid is more disordered than the same direction (upward stirring) of the three stirring paddles when the culture solution is stirred, and therefore, the stirring mode can greatly increase the dissolved oxygen amount, but also is easier to form turbulence due to the disordered impact of the liquid flow, and further cells are broken and dead. For this reason, the baffle plate 4 is arranged, the deflection angle of the baffle plate is consistent with the rotation direction of the stirring paddle, the baffle plate 4 can play a role in guiding flow, and the baffle plate 4 on the tank wall can slow down the flow velocity of the liquid at the position close to the tank wall of the bioreactor and guide the flow direction of the liquid because the flow velocity of the cell culture liquid at the position close to the tank wall of the bioreactor is the largest, so that the generation of turbulent flow is effectively avoided.
The invention also relates to a method for supplementing alkali in a bioreactor, which comprises intermittently opening the valve 501 to enable the alkali liquid injection head 5 to pulse the alkali liquid and simultaneously opening the stirring device 2 to enable the supplemented alkali liquid to be rapidly and completely mixed when the equipment is used for supplementing alkali.
Preferably, the volume of the tank body A is 3000L-6000L;
preferably, the method for supplementing alkali to the bioreactor is characterized in that the alkali solution is NaHCO of 0.8mol/L to 1.2mol/L 3 A solution; more preferably 1mol/L NaHCO 3 A solution.
Preferably, according to the method for supplementing alkali to the bioreactor, the valve 501 is opened once every 4.5 s-5.5 s, and each time is opened for 50 ms-200 ms; more preferably, the valve 501 is opened every 5 seconds, each time for 100ms to 150ms.
Preferably, according to the method for supplementing alkali in the bioreactor, when alkali liquid is sprayed, the liquid pressure in the alkali liquid spraying head 5 is 0.08-0.12 MPa; more preferably, the liquid pressure in the lye jet head 5 is 0.10MPa.
Preferably, in the method for supplementing alkali to a bioreactor as described above, the rotation speed of the stirring device 2 is 30rpm to 70rpm when the alkali supplementing operation is performed; more preferably, the rotation speed of the stirring device 2 is 50rpm.
The concentration of the alkali liquor, the switching time of the valve, the rotating speed of the stirring device and the pressure in the alkali liquor spraying head (the outlet diameter of the spraying head is matched, and the quantity of the alkali liquor sprayed out each time is practically limited) are specially set by the applicant in combination with the structure of the alkali supplementing equipment of the bioreactor so as to be capable of quickly uniformly mixing the alkali liquor and reduce the damage to cells.
The opening and closing of the valve and the rotating speed of the stirring device can be controlled by an electronic control device, and the control device reads and analyzes the data of the pH meter to further control the operation of the device.
Examples
In this embodiment, the bioreactor tank is a 6000L tank, and the suspension type MDCK cells are cultured. An alkali liquor spraying head is arranged above the tank wall and is connected with a storage tank through a valve by 1.0mol/LNaHCO 3 The containers of solution are connected and the valves are opened once every 5s, each time for 120ms. When the alkali liquor is sprayed, the pressure of the liquid in the alkali liquor spraying head is 1.0MPa, and the diameter of the spraying opening of the alkali liquor spraying head is 2mm. The bottom in the tank is provided with a microbubble ventilation device for ventilation of oxygen, the bottom in the tank is also provided with a stirring device, the stirring device comprises a stirring shaft and a stirring paddle, the stirring shaft is perpendicular to the horizontal plane and is fixed at the bottom of the tank, the stirring paddle is arranged on the stirring shaft, and the bottom of the stirring shaft is connected with a power device; the stirring shaft is provided with three stirring paddles, namely a first stirring paddle, a second stirring paddle and a third stirring paddle which are sequentially arranged from top to bottom, and the stirring shafts rotate clockwise; the second stirring paddle stirs the liquid to move from top to bottom; the first stirring paddles and the third stirring paddles stir the liquid to move from bottom to top. The blade inclination angle of the stirring paddle is 50 degrees, and the ratio of the paddle diameter to the tank diameter is 0.4. When the alkali solution is dripped, the rotating speed of the stirring paddle is 50 revolutions per minute; when the cells are normally cultured and oxygen is introduced, the rotating speed of the stirring paddle is 40 revolutions per minute. Four baffles are arranged on the wall of the tank, the deflection directions of the baffles are respectively clockwise deviated from the warp line 35 degrees where the baffles are positioned at the connecting point of the baffles and the tank body, and the width of the baffles is1/12 of the tank diameter. This example was set as an experimental group, on which a comparative example was set.
Comparative example 1: the only difference from the experimental group is the removal of the baffle.
Comparative example 2: removing the alkali liquor spraying head, and replacing an alkali adding mode by a side wall dropwise adding mode in the prior art; the rotation speed of the stirring paddle is always 40 revolutions per minute.
Comparative example 3: the only difference from the experimental group is that the baffle is arranged perpendicular to the tank.
Comparative example 4: the only difference from the experimental group is that all three paddles agitate the broth upward and remove the baffles.
The same batch of MDCK cells were cultured for 5 days at a cell culture density of 180 ten thousand/ml, and the pH was maintained constant at 6.8-7.2 by adding alkali, and the following data were examined:
as is clear from comparative examples 1 and 2, the different addition modes of the lye have a large influence on the cell viability. The wall-attached dripping mode is continuous, so that the concentration of alkali liquor at the dripping position is often overlarge, and more cells die. If the rotation speed is increased, the increase of the cell death rate caused by mechanical damage can be caused; in addition, the dosage of the wall-attached dripping is not controllable every time, and the pH cannot be accurately adjusted. As is evident from comparative examples 1 and 3, the baffle plate was not properly arranged or the baffle plate was removed, and although the amount of oxygen was reduced, more severe turbulence was formed, resulting in a significant decrease in the cell viability. As is clear from comparative example 4, when the culture solution is stirred up by all three stirring paddles, the residence time of oxygen in the culture solution is shortened, so that the oxygen utilization rate is reduced, and the total oxygen consumption is obviously increased; in addition, the change of the liquid stirring mode is not beneficial to uniform oxygen dispersion, and the difficulty of uniform mixing of oxygen is increased due to the adoption of the large-volume bioreactor, so that the local oxygen concentration is possibly reduced, and the cell activity is possibly reduced.
In summary, the invention adopts a pulse alkali liquor replenishing mode and is matched with a specially arranged stirring device and a baffle plate, so that turbulent flow generated during stirring can be effectively reduced, alkali liquor can be rapidly mixed, and cell death caused by mechanical damage and overhigh local alkali liquor concentration can be reduced. In addition, unexpectedly, the unique stirring paddle rotating mode can also prolong the residence time of the oxygen, thereby effectively reducing the consumption of the oxygen.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the invention.
Claims (9)
1. The alkali supplementing device of the bioreactor is characterized by comprising a tank body;
a stirring device is arranged in the tank body;
the stirring device comprises a stirring shaft and stirring paddles;
the stirring shaft is perpendicular to the horizontal plane and is fixed at the bottom of the tank, the stirring paddle is arranged on the stirring shaft, and the bottom of the stirring shaft is connected with the power device;
the stirring shafts are sequentially provided with a first stirring paddle, a second stirring paddle and a third stirring paddle from top to bottom; the second stirring paddle stirs the liquid to move from top to bottom; the first stirring paddles and the third stirring paddles stir the liquid to move from bottom to top;
the stirring paddle is a turbine type stirring paddle or a propelling type stirring paddle;
an alkali liquor spraying head is arranged above the tank wall of the tank body and is connected with an alkali storage container through a valve;
the device further comprises a micro-bubble ventilation device, wherein the micro-bubble ventilation device is arranged right below the stirring paddle;
a baffle is further arranged in the tank wall of the tank body, and the baffle is perpendicular to the horizontal plane;
the baffles deviate from the radial line of the connecting point of the baffles and the tank body by 30-40 degrees, and the deflection directions of the baffles are consistent;
the deflection angle of the baffle is consistent with the rotation direction of the stirring paddle;
the volume of the tank body is 3000L-6000L;
the alkali liquor is NaHCO with the concentration of 0.8mol/L to 1.2mol/L 3 A solution;
the valve is opened once every 4.5 s-5.5 s, and 50 ms-200 ms of the valve is opened every time;
when the alkali liquor is sprayed, the pressure of the liquid in the alkali liquor spraying head is 0.08-0.12 MPa;
and when the alkali supplementing operation is carried out, the rotating speed of the stirring device is 30-70 rpm.
2. The alkali supplementing device of the bioreactor according to claim 1, wherein the aperture of the inner hole at the front end of the alkali liquor spraying head is 1 mm-3 mm.
3. The bioreactor alkali replenishment apparatus of claim 1 wherein said paddles are four-bladed pusher paddles.
4. The bioreactor alkali replenishing device of claim 1, wherein the number of baffles is 3-5 and the baffles are uniformly distributed on the wall of the tank body.
5. The method for supplementing alkali in the bioreactor is characterized in that when the equipment of any one of claims 1-4 is used for supplementing alkali, the valve is opened intermittently, so that the alkali liquid is sprayed by the alkali liquid spraying head in a pulse mode, and meanwhile, the stirring device is opened, so that the supplemented alkali liquid is mixed rapidly and completely;
the tank body has a volume of 3000L-6000L.
6. The method for alkali supplementation in a bioreactor according to claim 5, whereinThe alkali liquor is NaHCO with the concentration of 0.8mol/L to 1.2mol/L 3 A solution.
7. The method for replenishing alkali in a bioreactor according to claim 5, wherein the valve is opened once every 4.5 s-5.5 s, and 50 ms-200 ms are opened every time.
8. The method for replenishing alkali in a bioreactor according to claim 5, wherein the liquid pressure in the alkali liquid spraying head is 0.08-0.12 MPa when alkali liquid is sprayed.
9. The method for alkali replenishment in a bioreactor according to claim 5, wherein the rotation speed of the stirring device is 30rpm to 70rpm when alkali replenishment is performed.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810073816.5A CN108048323B (en) | 2018-01-25 | 2018-01-25 | Alkali supplementing equipment and method for bioreactor |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810073816.5A CN108048323B (en) | 2018-01-25 | 2018-01-25 | Alkali supplementing equipment and method for bioreactor |
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| CN108048323A CN108048323A (en) | 2018-05-18 |
| CN108048323B true CN108048323B (en) | 2024-02-06 |
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| CN107619777A (en) * | 2016-12-09 | 2018-01-23 | 济南航晨生物科技有限公司 | A kind of high-viscosity products dedicated liquid fermentation tank |
| CN207958395U (en) * | 2018-01-25 | 2018-10-12 | 吉林冠界生物技术有限公司 | A kind of bioreactor benefit alkali equipment |
-
2018
- 2018-01-25 CN CN201810073816.5A patent/CN108048323B/en active Active
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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