CN114317203A - Fermentation cylinder suitable for pichia pastoris fermentation production usefulness - Google Patents
Fermentation cylinder suitable for pichia pastoris fermentation production usefulness Download PDFInfo
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- CN114317203A CN114317203A CN202011075420.8A CN202011075420A CN114317203A CN 114317203 A CN114317203 A CN 114317203A CN 202011075420 A CN202011075420 A CN 202011075420A CN 114317203 A CN114317203 A CN 114317203A
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Abstract
A fermentation tank suitable for fermentation production of pichia pastoris comprises a tank body and a stirring shaft vertically arranged in the tank body, wherein a power mechanism for driving the stirring shaft to rotate is fixedly arranged at the top of the tank body; an inclined blade type propulsion paddle is fixedly installed on the stirring shaft at the upper part of the tank body, an upper parabolic disc turbine paddle is fixedly installed on the stirring shaft at the middle part of the tank body, and a lower parabolic disc turbine paddle is fixedly installed on the stirring shaft at the lower part of the tank body; the upper parabolic disk turbine paddle and the lower parabolic disk turbine paddle respectively comprise disks fixedly sleeved on the stirring shaft, and at least 3 parabolic stirring paddles are arranged in the circumferential direction of the disks. This fermentation cylinder reasonable in design, simple and practical, impel the setting of oar, last parabola disk turbine oar and lower parabola disk turbine oar through oblique leaf formula, can effectively strengthen gas-liquid exchange, the dwell time of extension air in the zymotic fluid can fine satisfy the oxygen suppliment and the mass transfer heat transfer demand of pichia pastoris in high density fermentation.
Description
Technical Field
The invention relates to a fermentation tank, in particular to a fermentation tank suitable for fermentation production of pichia pastoris.
Background
Pichia pastoris is an expression system capable of efficiently expressing foreign proteins, has the advantages of stable heredity, high expression level, capability of post-translational processing of proteins, capability of secreting products, capability of high-density fermentation and the like, and is widely applied, so that the pichia pastoris is commonly used for expressing the commonly used foreign proteins.
The high-density fermentation is an important strategy for improving the expression level of foreign proteins by genetically engineered bacteria, in the process of high-density fermentation, mature equipment needs to provide sufficient oxygen supply, carbon sources, nitrogen sources, inorganic salts and other nutrient substances, the oxygen supply condition is closely related to the size design, air flow, pressure and the like of the equipment, the good oxygen supply condition and the mixing of a culture medium are necessary conditions for ensuring heat transfer and mass transfer in the fermentation process, the design of an oxygen supply structure of a fermentation tank suitable for producing specific strains and products is a set of fermentation tanks, and the high-density fermentation has great significance for improving the fermentation yield, saving energy and reducing consumption.
Disclosure of Invention
The invention aims to solve the technical problem of the prior art and provides a fermentation tank which is reasonable in design, practical and convenient, can well meet the requirements of oxygen supply and mass and heat transfer of pichia pastoris in high-density fermentation, and is suitable for fermentation production of pichia pastoris.
The technical problem to be solved by the present invention is achieved by the following technical means. The invention relates to a fermentation tank suitable for fermentation production of pichia pastoris, which comprises a tank body and a stirring shaft vertically arranged in the tank body, wherein the top of the tank body is also fixedly provided with a power mechanism for driving the stirring shaft to rotate; an inclined blade type propulsion paddle is fixedly installed on the stirring shaft at the upper part of the tank body, an upper parabolic disc turbine paddle is fixedly installed on the stirring shaft at the middle part of the tank body, and a lower parabolic disc turbine paddle is fixedly installed on the stirring shaft at the lower part of the tank body; the upper parabolic disk turbine paddle and the lower parabolic disk turbine paddle respectively comprise disks fixedly sleeved on the stirring shaft, and at least 3 parabolic stirring paddles are arranged in the circumferential direction of the disks.
The technical problem to be solved by the invention can be further solved by adopting the following technical scheme that the inclined blade type propulsion paddle comprises 6 rectangular stirring blades, the 6 rectangular stirring blades are uniformly distributed along the circumferential direction of the stirring shaft, an inclined stirring surface is arranged at the end part of each stirring blade far away from the stirring shaft, and the upward part of each stirring surface inclines towards one side of the stirring shaft.
The technical problem to be solved by the invention can be further realized by the following technical scheme that the inclination angle of the stirring surface is 45-60 degrees for the fermentation tank suitable for pichia pastoris fermentation production.
The technical problem to be solved by the invention can be further realized by the following technical scheme that for the fermentation tank suitable for fermentation production of pichia pastoris, the disks of the upper parabolic disk turbine paddle and the lower parabolic disk turbine paddle are respectively provided with 6 parabolic stirring paddles, and the 6 parabolic stirring paddles are uniformly distributed along the circumferential direction of the disks.
The technical problem to be solved by the invention can be further realized by the following technical scheme that for the fermentation tank suitable for pichia pastoris fermentation production, the lengths of the inclined blade type propulsion paddle, the upper parabolic disk turbine paddle and the lower parabolic disk turbine paddle are 1/2-2/3 of the diameter of the tank body.
The technical problem to be solved by the invention can be further realized by the following technical scheme that for the fermentation tank suitable for pichia pastoris fermentation production, the power mechanism is a driving motor, the driving motor is fixedly arranged at the top of the tank body, and the top of the stirring shaft extends out of the tank body and is in transmission connection with an output shaft of the driving motor.
The technical problem to be solved by the invention can be further realized by the following technical scheme that the tank body is integrally cylindrical for the fermentation tank suitable for fermentation production of pichia pastoris.
Compared with the prior art, the invention arranges the stirring shaft in the tank body, and the inclined blade type propulsion paddle, the upper parabolic disk turbine paddle and the lower parabolic disk turbine paddle are sequentially arranged on the stirring shaft from top to bottom, the middle of the upper parabolic disk turbine paddle and the lower parabolic disk turbine paddle is a disk, and the periphery of the disk is provided with 6 parabolic stirring paddles; after air is introduced from the bottom of the fermentation tank, the air is distributed to the periphery due to the blocking of the disc of the lower parabolic disc turbine paddle and is uniformly distributed to the periphery under the wrapping and stirring of the parabolic stirring paddle; the arrangement of the upper parabolic disc turbine paddle avoids the condition of short circuit between air and the middle, and meanwhile, the heat and mass transfer in the fermentation liquor is effectively improved; meanwhile, the stirring paddles of the upper parabolic disk turbine paddle and the lower parabolic disk turbine paddle are parabolic, so that the reduction of liquid resistance is facilitated; and secondly, when the fermentation liquor reaches the upper part of the fermentation tank, the fermentation liquor moves downwards to the periphery under the action of the inclined blade type propulsion paddle, so that gas-liquid exchange is enhanced, the retention time of air in the fermentation liquor is prolonged, and the mass and heat transfer capacity of the tank body is improved. This fermentation cylinder reasonable in design, simple and practical, impel the setting of oar, last parabola disk turbine oar and lower parabola disk turbine oar through oblique leaf formula, can effectively strengthen gas-liquid exchange, the dwell time of extension air in the zymotic fluid can fine satisfy the oxygen suppliment and the mass transfer heat transfer demand of pichia pastoris in high density fermentation.
Drawings
FIG. 1 is a schematic diagram of a structure of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings of the present invention, and it is obvious that the described embodiments are some embodiments of the present invention, but not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Referring to fig. 1, the fermentation tank suitable for pichia pastoris fermentation production comprises a tank body 1 and a stirring shaft 2 vertically installed in the tank body 1, wherein the tank body 1 is vertically arranged, the lower part of the tank body 1 is communicated with an air duct 6, and one end of the air duct 6 extends to the bottom inside the tank body 1, so that air can be conveniently introduced into the tank body 1 from the bottom of the tank body 1; the stirring shaft 2 is used for stirring and pushing the fermentation liquor to flow and mix with air, so that the gas-liquid contact area is increased, the dissolved oxygen content in the fermentation liquor is increased, and the requirement of high-density culture of thalli is met; a power mechanism for driving the stirring shaft 2 to rotate is fixedly arranged at the top of the tank body 1 and is used for driving the stirring shaft 2 to rotate; an inclined blade type propulsion paddle 3 is fixedly arranged on the stirring shaft 2 at the upper part of the tank body 1 and is used for stirring the fermentation liquor at the upper part of the tank body 1, so that the fermentation liquor flows downwards to the periphery; an upper parabolic disc turbine impeller 4 is fixedly arranged on the stirring shaft 2 in the middle of the tank body 1 and is used for stirring in the middle of the tank body 1, so that air in the middle of the tank body 1 is diffused to the periphery, and the condition that the air is short-circuited in the middle is avoided; a lower parabolic disc turbine impeller 5 is fixedly arranged on the stirring shaft 2 at the lower part of the tank body 1 and is used for stirring at the lower part of the tank body 1, so that the air introduced into the tank body 1 is distributed to the periphery; the upper parabolic disk turbine impeller 4 and the lower parabolic disk turbine impeller 5 respectively comprise disks fixedly sleeved on the stirring shaft 2, and at least 3 parabolic stirring impellers are arranged in the circumferential direction of the disks; the disc can be used for blocking air, and the stirring paddle is in a parabolic shape, so that the air is conveniently stirred, and the liquid resistance of the fermentation liquid is favorably reduced.
The inclined blade type propulsion paddle 3 comprises 6 rectangular stirring blades, the 6 rectangular stirring blades are uniformly distributed along the circumferential direction of the stirring shaft 2, and the angle between every two adjacent rectangular stirring blades is 60 degrees; the end part of the stirring blade far away from the stirring shaft 2 is provided with an inclined stirring surface, the upward part of the stirring surface inclines towards one side of the stirring shaft 2, so that the fermentation liquor can move downwards to the periphery through stirring.
The inclination angle of the stirring surface is 45-60 degrees, preferably, the inclination angle of the stirring surface is 60 degrees, fermentation liquor can better move downwards and around through stirring, gas-liquid exchange is enhanced, and the retention time of air in the fermentation liquor is prolonged.
The discs of the upper parabolic disc turbine paddle 4 and the lower parabolic disc turbine paddle 5 are respectively provided with 6 parabolic-shaped stirring paddles, the 6 parabolic-shaped stirring paddles are uniformly distributed along the circumferential direction of the discs, and the angle between every two adjacent parabolic-shaped stirring paddles is 60 degrees, so that fermentation liquor can be stirred, the mass and heat transfer efficiency of the fermentation liquor is improved, and the liquid resistance of the fermentation liquor during stirring can be reduced.
The lengths of the inclined blade type propulsion paddle 3, the upper parabolic disk turbine paddle 4 and the lower parabolic disk turbine paddle 5 are 1/2-2/3 of the diameter of the tank body 1. Preferably, the lengths of the blade type propulsion paddle, the upper parabolic disk turbine paddle 4 and the lower parabolic disk turbine paddle 5 are 2/3 of the diameter of the tank body 1, so that large-scale stirring can be performed in the tank body 1, high-density culture of pichia pastoris is facilitated, and the oxygen and mass transfer requirements of the pichia pastoris are met.
The power mechanism is a driving motor which is fixedly arranged at the top of the tank body 1, and the top of the stirring shaft 2 extends out of the tank body 1 and is in transmission connection with an output shaft of the driving motor. A transmission gear is fixedly installed on an output shaft of a driving motor, a fixed gear meshed with the transmission gear is fixedly installed on a stirring shaft 2, the driving motor is started, the output shaft of the driving motor rotates, the stirring shaft 2 is driven to rotate through the transmission gear and the fixed gear matched in transmission, and then an inclined blade type propulsion paddle 3, an upper parabolic disk turbine paddle 4 and a lower parabolic disk turbine paddle 5 can be driven to rotate, so that fermentation liquor in the fermentation tank is stirred.
The tank body 1 is integrally cylindrical and convenient to manufacture and use, and a support is fixedly arranged at the bottom of the tank body 1 and used for being matched with the tank body 1 to install.
For the lower part of the tank body 1, due to the arrangement of the lower parabolic disk turbine paddle 5, after air is introduced from the bottom of the fermentation tank, the air is distributed to the periphery due to the blocking of the disk of the lower parabolic disk turbine paddle 5, and then is uniformly distributed to the periphery under the wrapping and stirring of the stirring paddle, and meanwhile, the stirring paddle is in a parabolic shape, which is beneficial to reducing the liquid resistance;
for the middle part of the tank body 1, when the liquid level in the tank just submerges the upper parabolic disk turbine paddle 4 in the early stage of fermentation, the liquid level occupies about 1/3 of the total height of the fermentation tank, and the distance between the upper parabolic disk turbine paddle 4 and the lower parabolic disk turbine paddle 5 is short, so that the arrangement of the lower parabolic disk turbine paddle 5 avoids the situation of short circuit of air passing through the middle, and meanwhile, the heat and mass transfer in the fermentation liquid is effectively improved;
for the upper part of the tank body 1, in the period of expressing the phospholipase C by pichia pastoris, due to the influence of feeding and fermentation expression protein, the liquid level of fermentation liquor floods the inclined-blade type propulsion paddle 3, at the position, most of oxygen in the air entering from the bottom is consumed, and partial thalli of the middle and upper layer fermentation liquor can not obtain sufficient oxygen, and the arrangement of the inclined-blade type propulsion paddle 3 enables the fermentation liquor to move downwards to the periphery, so that gas-liquid exchange is enhanced, the retention time of the air in the fermentation liquor is prolonged, and the whole mass and heat transfer capacity of the tank body 1 is improved.
The fermentation tank provided by the application is applied in practice, in the production process of a 5m fermentation tank for expressing phospholipase C by pichia pastoris and under the same fermentation condition, indexes such as the highest OUR (thallus oxygen uptake rate), the thallus wet weight, the fermentation protein yield, the fermentation enzyme activity and the like in the fermentation process are all improved, wherein the yield is improved by 32% to the maximum extent, and the good effects of saving energy, reducing consumption and reducing cost are achieved; specific data are shown in table 1:
TABLE 1
Claims (7)
1. The utility model provides a fermentation cylinder suitable for pichia pastoris fermentation production usefulness which characterized in that: the fermentation tank comprises a tank body and a stirring shaft vertically arranged in the tank body, and a power mechanism for driving the stirring shaft to rotate is also fixedly arranged at the top of the tank body; an inclined blade type propulsion paddle is fixedly installed on the stirring shaft at the upper part of the tank body, an upper parabolic disc turbine paddle is fixedly installed on the stirring shaft at the middle part of the tank body, and a lower parabolic disc turbine paddle is fixedly installed on the stirring shaft at the lower part of the tank body; the upper parabolic disk turbine paddle and the lower parabolic disk turbine paddle respectively comprise disks fixedly sleeved on the stirring shaft, and at least 3 parabolic stirring paddles are arranged in the circumferential direction of the disks.
2. The fermentation tank suitable for pichia pastoris fermentation production according to claim 1, wherein: the oblique blade type propulsion paddle comprises 6 rectangular stirring blades, the 6 rectangular stirring blades are uniformly distributed along the circumferential direction of the stirring shaft, an oblique stirring surface is arranged at the end part, far away from the stirring shaft, of each stirring blade, and the stirring surface inclines towards one side of the stirring shaft towards the upper part.
3. The fermentation tank suitable for pichia pastoris fermentation production according to claim 2, wherein: the inclination angle of the stirring surface is 45-60 degrees.
4. The fermentation tank suitable for pichia pastoris fermentation production according to claim 1, wherein: the disks of the upper parabolic disk turbine paddle and the lower parabolic disk turbine paddle are respectively provided with 6 stirring paddles in parabolic shapes, and the 6 stirring paddles in parabolic shapes are uniformly distributed along the circumferential direction of the disks.
5. The fermentation tank suitable for pichia pastoris fermentation production according to claim 1, wherein: the lengths of the inclined blade type propulsion paddle, the upper parabolic disk turbine paddle and the lower parabolic disk turbine paddle are 1/2-2/3 of the diameter of the tank body.
6. The fermentation tank suitable for pichia pastoris fermentation production according to claim 1, wherein: the power mechanism is a driving motor which is fixedly arranged at the top of the tank body, and the top of the stirring shaft extends out of the tank body and is in transmission connection with an output shaft of the driving motor.
7. The fermentation tank suitable for pichia pastoris fermentation production according to claim 1, wherein: the tank body is integrally cylindrical.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011075420.8A CN114317203A (en) | 2020-10-09 | 2020-10-09 | Fermentation cylinder suitable for pichia pastoris fermentation production usefulness |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011075420.8A CN114317203A (en) | 2020-10-09 | 2020-10-09 | Fermentation cylinder suitable for pichia pastoris fermentation production usefulness |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114317203A true CN114317203A (en) | 2022-04-12 |
Family
ID=81032183
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011075420.8A Withdrawn CN114317203A (en) | 2020-10-09 | 2020-10-09 | Fermentation cylinder suitable for pichia pastoris fermentation production usefulness |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114317203A (en) |
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2020
- 2020-10-09 CN CN202011075420.8A patent/CN114317203A/en not_active Withdrawn
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Application publication date: 20220412 |