CN220724010U - Recombinant collagen separation and purification system - Google Patents

Abstract

The utility model provides a recombinant collagen separation and purification system which comprises a bacterial liquid separation subsystem, a first separation subsystem, a second separation subsystem, a filtering subsystem and a material receiving subsystem. The system is additionally provided with a bacterial liquid separation subsystem, so that fermented feed liquid can be separated into bacterial liquid and bacterial liquid, and meanwhile, recombinant collagen with different expression forms can be respectively discharged into different separation subsystems according to different reagents and treatment methods, so that different extraction and purification treatments can be conveniently carried out, further more recombinant collagen can be ensured to be obtained, and the protein extraction efficiency is improved. The feed liquid treated by the separation system is subjected to multi-layer filtration treatment, so that the purity of the product is ensured. In addition, extracellular expression recombinant collagen does not need thallus fragmentation, so that the extraction process is simplified; the recombinant collagen bacteria which are expressed in the cell and simultaneously expressed in the cell and the cell need to be crushed, and only need to be purified in 2-3 steps, so that the purification steps are simplified, and the treatment efficiency is improved.

Description

Recombinant collagen separation and purification system

Technical Field

The utility model belongs to the technical field of separation and purification, and relates to a recombinant collagen separation and purification system.

Background

Collagen is the main component of extracellular matrix, accounting for 70% -80% of dry weight in human tissue. The collagen mainly comprises type I and type III collagen, wherein the collagen in the skin of the infant is mainly type III collagen, and the type III collagen accounts for 60% of the total collagen; collagen in skin of young and old people is mainly type I collagen, and type I collagen accounts for 80% of total collagen. Type I collagen determines skin contour support, and is a material basis for scar tissue fibrosis; type III collagen is the main component of reticular fibers in the skin, and determines the elasticity and fineness of the skin. The higher the content of type III collagen, the finer the fiber bundles. In wound repair, the skin tissue is fine and smooth when the proportion of III type collagen is high.

Collagen is mainly synthesized by fibroblasts, but adult fibroblasts of their own dermis can only synthesize type I collagen. In the growth process of the human, the proportion of the III type collagen is gradually reduced, and the proportion of the I type collagen is continuously increased. Type I and iii collagen levels are closely related to skin youthfulness, wound repair, cell support and regeneration, and the like. Symptoms such as epithelial damage and skin atrophy are accompanied by a large loss of collagen, so that the main way to repair the skin is to supplement collagen in a large amount. Currently, the collagen used in the market is mainly type I collagen, the active structure of which is often destroyed, and there is a risk of contamination and allergy of animal-derived viruses.

The recombinant collagen is recombinant human-derived collagen which is obtained by using a genetic engineering technology and taking a human collagen original gene sequence as a template and expressing the recombinant human-derived collagen in escherichia coli or saccharomycetes through optimized recombination and has high biocompatibility and high activity and is highly consistent with the human collagen. Compared with natural collagen, the recombinant collagen is rich in hydrophilic amino acid residues, has excellent water solubility, higher biological activity, no virus hidden trouble, high biocompatibility and low immune rejection, and can promote cell adhesion and proliferation and differentiation.

Recombinant collagen is stored in yeast cells or bacteria after expression, secreted extracellularly, or expressed in yeast cells or bacteria at the same time. During extraction of intracellular expression recombinant collagen, the bacterial cells need to be crushed and then purified to obtain the recombinant collagen. In the extraction of the extracellular recombinant collagen, the cell is not required to be broken. Therefore, two sets of devices are needed for separation and purification of the recombinant collagen in two expression modes, and separation and purification cannot be realized on the same device.

Disclosure of Invention

The utility model aims to provide a recombinant collagen separation and purification system, which aims to solve the problem that the separation and purification of recombinant collagen in two expression modes cannot be realized on the same device.

In order to achieve the above purpose, the present utility model adopts the following technical scheme:

the utility model provides a recombinant collagen separation and purification system, which comprises a bacterial liquid separation subsystem, a first separation subsystem and a second separation subsystem, wherein the first separation subsystem and the second separation subsystem are all communicated with the bacterial liquid separation subsystem, the first separation subsystem is communicated with a filtering subsystem through a first conveying pipe, the second separation subsystem is communicated with the filtering subsystem through a second conveying pipe, and the bottom of the filtering subsystem is also communicated with a receiving subsystem; the bacterial liquid separation subsystem comprises a liquid storage tank, a separation rotating motor arranged on the outer side of the top of the liquid storage tank and a separator arranged in the liquid storage tank; a separation rotating shaft is arranged at the top of the inner side of the separator, and the separation rotating shaft is connected with the separation rotating motor; a separation filter screen is arranged on the side wall of the separator, a rubber plug, a cam and a driving motor are arranged at the bottom of the separator, the rubber plug is movably plugged at the bottom of the separator, and the cam is respectively connected with the rubber plug and the driving motor; the top of liquid storage pot is equipped with first inlet and second inlet, first inlet intercommunication the inside of separator, the second inlet intercommunication the inside of liquid storage pot.

Preferably, the first separation subsystem comprises a first separation tank, a first rotating motor arranged outside the first separation tank, a first separation tank feed inlet arranged at the top of the first separation tank and a first separation tank discharge outlet arranged at the bottom of the first separation tank; the first separating tank feed inlet is communicated with the bacterial liquid separating subsystem, and the first separating tank discharge outlet is communicated with the first conveying pipe; the first rotary shaft is arranged inside the first separating tank, rotary separating blades are arranged on the first rotary shaft, and the first rotary shaft is connected with the first rotary motor.

Preferably, the second separation system comprises a second separation tank, a second rotating motor arranged outside the second separation tank, a second separation tank feed inlet and a liquid adding port arranged at the top of the second separation tank, and a second separation tank discharge outlet arranged at the bottom of the second separation tank; the feed inlet of the second separation tank is communicated with the bacterial liquid separation subsystem, and the discharge outlet of the second separation tank is communicated with the second conveying pipe; the inside second rotation axis that sets up of second knockout drum, set up crushing separation blade on the second rotation axis, the second rotation axis is connected the second rotating electrical machines.

Preferably, the filtering subsystem comprises a first-stage filtering chamber, a second-stage filtering chamber, a third-stage filtering chamber and a fourth-stage filtering chamber which are sequentially arranged from top to bottom, wherein the first-stage filtering chamber is communicated with the second conveying pipe, the second-stage filtering chamber is communicated with the first conveying pipe, and the fourth-stage filtering chamber is communicated with the material receiving subsystem; a filtering device is movably arranged between two adjacent filtering chambers.

Preferably, the filtering device comprises a filter membrane carrier plate, a filter membrane arranged on the filter membrane carrier plate and a filter membrane cover plate covering the filter membrane, and the filter membrane cover plate is provided with a through hole.

Preferably, the diameter of the mesh on the filter membrane gradually decreases from the first stage filter chamber to the fourth stage filter chamber.

Preferably, a handle is arranged on the outer side of the filter membrane carrier plate.

Preferably, the material receiving subsystem comprises a material receiving tank and a material receiving pipe, and two ends of the material receiving pipe are respectively communicated with the material receiving tank and the filtering subsystem.

Preferably, a first conveying pump is arranged on the first conveying pipe, and a second conveying pump is arranged on the second conveying pipe.

Preferably, the system further comprises a fixed support, the bacterial liquid separation subsystem and the filtering subsystem are located above the fixed support, and the first separation subsystem, the second separation subsystem and the material receiving subsystem are located below the fixed support.

The utility model has the following beneficial effects:

the utility model provides a recombinant collagen separation and purification system which comprises a bacterial liquid separation subsystem, a first separation subsystem, a second separation subsystem, a filtering subsystem and a material receiving subsystem. The system is additionally provided with a bacterial liquid separation subsystem, so that fermented feed liquid can be separated into bacterial liquid and bacterial liquid, and meanwhile, recombinant collagen with different expression forms can be respectively discharged into different separation subsystems according to different reagents and treatment methods, so that different extraction and purification treatments can be conveniently carried out, further more recombinant collagen can be ensured to be obtained, and the protein extraction efficiency is improved. The feed liquid treated by the separation system is subjected to multi-layer filtration treatment, so that the purity of the product is ensured. In addition, extracellular expression recombinant collagen does not need thallus fragmentation, so that the extraction process is simplified; the recombinant collagen bacteria which are expressed in the cell and simultaneously expressed in the cell and the cell need to be crushed, and only need 2-3 steps of purification treatment, so that the purification steps are simplified, and the treatment efficiency is improved.

Drawings

Fig. 1 is a schematic perspective view of a recombinant collagen separation and purification system according to an embodiment of the present application;

FIG. 2 is a front view of a recombinant collagen separation and purification system according to an embodiment of the present disclosure;

FIG. 3 is a cross-sectional view of a bacterial liquid separation system according to an embodiment of the present disclosure;

FIG. 4 is a block diagram of a first separation subsystem provided in an embodiment of the present application;

FIG. 5 is a block diagram of a second separation system according to an embodiment of the present disclosure;

FIG. 6 is a block diagram of a filtering subsystem provided in an embodiment of the present application;

fig. 7 is a perspective view of a filtering device according to an embodiment of the present disclosure;

FIG. 8 is a block diagram of a material receiving subsystem according to an embodiment of the present disclosure;

the symbols represent:

01-bacterial liquid separation subsystem, 02-first separation subsystem, 03-second separation subsystem, 04-first conveying pipe, 05-filtering subsystem, 06-second conveying pipe, 07-receiving subsystem, 08-first conveying pump, 09-second conveying pump and 010-fixed bracket;

1-a liquid storage tank, 2-a separation rotating motor, 3-a separator, 4-a separation rotating shaft, 5-a rubber plug, 6-a cam, 7-a driving motor, 8-a first liquid inlet, 9-a second liquid inlet, 10-a first separation tank, 11-a first rotating motor, 12-a first separation tank feed inlet, 13-a first separation tank discharge outlet, 14-a first rotating shaft, 15-a rotary separation blade, 16-a second separation tank, 17-a second rotating motor, 18-a second separation tank feed inlet, 19-a liquid feeding port, 20-a second separation tank discharge outlet, 21-a second rotating shaft, 22-a crushing separation blade, 23-a first stage filter chamber, 24-a second stage filter chamber, 25-a third stage filter chamber, 26-a fourth stage filter chamber, 27-a filter device, 28-a handle, 29-a receiving tank, 30-receiving pipe;

2701-filter membrane carrier plate, 2702-filter membrane, 2703-filter membrane cover plate and 2704-through holes.

Detailed Description

The following description of the embodiments of the present utility model will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present utility model, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the utility model without making any inventive effort, are intended to be within the scope of the utility model.

Referring to fig. 1 and 2, fig. 1 and 2 show a schematic perspective view and a front view of a recombinant collagen separation and purification system according to an embodiment of the present application. As can be seen from fig. 1, the recombinant collagen separation and purification system provided in the embodiment of the present application includes: cell bacterial liquid separation subsystem 01, first separation subsystem 02, second separation subsystem 03, first conveying pipe 04, filtering subsystem 05, second conveying pipe 06, material receiving subsystem 07 and fixing support 010. Wherein, the fixed support 010 is a support component of a recombinant collagen separation and purification system, and consists of a baffle plate and supporting legs. The bacterial liquid separation subsystem 01 and the filtering subsystem 05 are arranged above the fixed support 010, and the first separation subsystem 02, the second separation subsystem 03, the first conveying pipe 04, the second conveying pipe 06 and the material receiving subsystem 07 are all arranged below the fixed support 010.

The bacterial liquid separation subsystem 01 is a component for separating bacterial liquid and bacterial liquid, and is respectively communicated with the first separation subsystem 02 and the second separation subsystem 03, so that the separated bacterial liquid and bacterial liquid are respectively conveyed into the first separation subsystem 02, the second separation subsystem 03 or the waste liquid collecting device. The first separation subsystem 02 and the second separation subsystem 03 are components for stirring and crushing thalli containing recombinant collagen with different expression modes, wherein the first separation subsystem 02 is communicated with the filtering subsystem 05 through a first conveying pipe 04, the second separation subsystem 03 is communicated with the filtering subsystem 05 through a second conveying pipe 06, and therefore, thalli containing recombinant collagen with different expression modes are separated through different devices, and crushed thalli are conveyed into the filtering subsystem 05 through different pipelines respectively. The filtering subsystem 05 filters the crushed thalli discharged to obtain recombinant collagen, and the recombinant collagen is collected in the material receiving subsystem 07. Because the first separation subsystem 02 and the second separation subsystem 03 are positioned below the fixed support 010, and the filtering subsystem 05 is positioned above the fixed support 010, in order to facilitate the conveying of materials, the first conveying pipe 04 is provided with a first conveying pump 08, and the second conveying pipe 06 is provided with a second conveying pump 09.

The bacterial cell liquid separation subsystem 01, the first separation subsystem 02, the second separation subsystem 03, the filtration subsystem 05 and the material receiving subsystem 07 in the embodiment of the present application will be described in detail below with reference to the accompanying drawings.

Referring to fig. 3, fig. 3 is a cross-sectional view of a bacterial liquid separation subsystem 01 according to an embodiment of the present application. As can be seen from fig. 3, the bacterial liquid separation subsystem 01 in the embodiment of the present application includes a liquid storage tank 1, a separation rotary motor 2, a separator 3, and the like.

Specifically, the liquid storage tank 1 is a main body part of the bacterial liquid separation subsystem 01, and is arranged on the fixed bracket 010 through four supporting legs. The inside of the liquid storage tank 1 is vertically provided with a separator 3, and the separator 3 is used for separating bacterial liquid. The separator 3 is a hollow member, and a separation rotation shaft 4 is vertically provided at the top of the inner side thereof. A separation rotating motor 2 is arranged on the outer side of the top of the liquid storage tank 1, and the separation rotating motor 2 is connected with a separation rotating shaft 4. Therefore, when the separation rotary motor 2 rotates, the separation rotary shaft 4 can be driven to rotate, so that the separator 3 rotates, and separation of bacterial liquid of the bacterial cells is realized under the action of centrifugal force. The side wall of the separator 3 is provided with a separation hole which is used for filtering the separated bacterial liquid to realize the separation of solid precipitate bacterial liquid and liquid material bacterial liquid.

The bottom of the separator 3 is provided with a rubber plug 5, a cam 6 and a driving motor 7, and the rubber plug 5 is movably plugged at the bottom of the separator 3, and the cam 6 is respectively connected with the rubber plug 5 and the driving motor 7. Under the drive of the driving motor 7, the cam 6 can rotate and drive the rubber plug 5 to move up and down, so that the rubber plug 5 plugs the bottom of the separator 3 and is separated from the separator 3.

The top of liquid storage pot 1 is equipped with first inlet 8 and second inlet 9, and wherein, first inlet 8 communicates the inside of separator 3, and second inlet 9 communicates the inside of liquid storage pot 1. The first liquid inlet 8 is used for feeding liquid containing bacterial liquid, and the second liquid inlet 9 is used for feeding liquid, reagent and cleaning liquid.

In the bacterial liquid separation subsystem 01, after a feed liquid containing bacterial liquid enters the separator 3 from the first liquid inlet 8, the separation rotary motor 2 is started, the separation rotary shaft 4 is driven to rotate, and bacterial liquid is separated to form bacterial liquid and bacterial liquid. The thalli is collected at the bottom of the inside of the separator 3 as solid materials, and the bacterial liquid enters between the separator 3 and the liquid storage tank 1 as liquid materials through separation holes on the side wall of the separator 3. The driving motor 7 is started, so that the driving motor 7 drives the cam 6 to rotate, and further drives the rubber plug 5 to move up and down. When the rubber stopper 5 moves downward, the rubber stopper 5 is separated from the separator 3, and at this time, the bottom of the separator 3 is in an open state, and the cells located at the bottom inside the separator 3 are discharged into the second separation system 03. The bacterial liquid is discharged to the first separation subsystem 02 or the waste liquid collecting device through a liquid outlet according to the situation. The liquid for cleaning cleans the liquid storage tank 1 and the separator 3 through the second liquid inlet 9. The reagent used for extracting and separating the recombinant collagen is discharged into the liquid storage tank 1 through the second liquid inlet 9, and then is discharged into the first separation subsystem 02 through the liquid outlet.

The separation subsystem in the present application is divided into two parts, namely, a first separation subsystem 02 and a second separation subsystem 03, which can stir the discharged thalli and the liquid or liquid reagent decomposing the thalli, or break up and stir the thalli, respectively. The first separation subsystem 02 is used for stirring bacterial liquid and a reagent so as to realize extraction of extracellular recombinant collagen; the second separation system 03 is used for crushing and stirring the thalli and the reagents for decomposing the thalli so as to realize extraction of the recombinant collagen expressed in the cells.

As can be seen from fig. 4, the first separation subsystem 02 provided in the embodiment of the present application includes a first separation tank 10, and the first separation tank 10 is a tank structure. The outside of first knockout drum 10 is equipped with first rotating electrical machines 11, and the top is equipped with first knockout drum feed inlet 12, and the bottom is equipped with first knockout drum discharge gate 13, and wherein, first knockout drum feed inlet 12 intercommunication thallus fungus liquid separation subsystem 01, first knockout drum discharge gate 13 intercommunication first conveyer pipe 04 to in being convenient for carry the material of separating to filtration subsystem 05 through first conveyer pipe 04.

The first rotary shaft 14 is provided in the first separator tank 10, the rotary separation blades 15 are provided on the first rotary shaft 14, and the first rotary shaft 14 is connected to the first rotary motor 11. When the first rotary motor 11 is started, the first rotary shaft 14 is rotated by the driving action of the first rotary motor 11, and thereby drives the rotary separation blade 15 to rotate. The rotation of the rotary separation blade 15 can stir the bacterial liquid and the reagent, so that the reagent is uniformly distributed in the bacterial liquid, and the phenomenon that substances which cannot be filtered during filtration suddenly increase can not occur, thereby preventing blockage in the filtration process.

As can be seen from fig. 5, the second separation system 03 provided in the embodiment of the present application includes a second separation tank 16, and the second separation tank 16 is also a tank structure. The outside of second knockout drum 16 is equipped with second rotating electrical machines 17, and the top is equipped with second knockout drum feed inlet 18 and liquid filling opening 19, and the bottom is equipped with second knockout drum discharge gate 20, and wherein, second knockout drum feed inlet 18 intercommunication thallus fungus liquid separation subsystem 01, second knockout drum discharge gate 20 intercommunication second conveyer pipe 06 to in being convenient for carry the material of separating to filtration subsystem 05 through second conveyer pipe 06.

The second rotary shaft 21 is provided in the second separation tank 16, the crushing separation blades 22 are provided on the second rotary shaft 21, and the second rotary shaft 21 is connected to the second rotary motor 17. When the second rotary motor 17 is started, the second rotary shaft 21 is rotated by the driving of the second rotary motor 17, and thereby drives the crushing and separating blades 22 to rotate. After the cells enter the second separation tank 16, the lysate is added through the liquid inlet 19. Under the rotation of the crushing and separating blades 22, the crushing and separating blades 22 crush and agitate the bacterial cells. In the crushing and stirring processes, the thalli are rapidly crushed by physical stirring and a lysate biological enzymolysis method, so that the wall breaking effect is enhanced, and the release efficiency of the recombinant collagen is further improved. The thallus is crushed and stirred by the crushing and separating blades 22, so that the thallus and the liquid for decomposing the thallus or the liquid reagent and the pyrolysis liquid are uniformly distributed, and the blockage in the filtering process is prevented; but also can improve the extraction efficiency of the recombinant collagen.

The materials after the first separation subsystem 02 and the second separation subsystem 03 are stirred and crushed are respectively conveyed into the filtering subsystem 05 through the first conveying pipe 04 and the second conveying pipe 06, and then the materials are filtered by the filtering subsystem 05, so that the purification of recombinant proteins is realized.

As can be seen from fig. 6, the filtering subsystem 05 provided in the embodiment of the present application includes a first stage filtering chamber 23, a second stage filtering chamber 24, a third stage filtering chamber 25 and a fourth stage filtering chamber 26, which are sequentially arranged from top to bottom; wherein the first stage filtering chamber 23 is communicated with the second conveying pipe 06 and is used for receiving the materials separated by the second separation subsystem 03; the second stage filter chamber 24 is communicated with the first conveying pipe 04 and is used for receiving materials separated by the first separation subsystem 02; the fourth stage filter chamber 26 is communicated with the material receiving subsystem 07 and is used for collecting the filtered recombinant collagen into the material receiving subsystem 07.

A filtering device 27 is movably arranged between two adjacent filtering chambers, namely, a first-stage filtering chamber 23 and a second-stage filtering chamber 24, a second-stage filtering chamber 24 and a third-stage filtering chamber 25, and a filtering device 27 is respectively arranged between the third-stage filtering chamber 25 and a fourth-stage filtering chamber 26. The filter device 27 in this embodiment of the present application includes a filter membrane carrier plate 2701, a filter membrane 2702, and a filter membrane cover plate 2703, as shown in fig. 7. The filter membrane carrier 2701 has a frame-type plate structure, and a filter membrane 2702 is provided therein. The filter 2702 is covered with a filter cover plate 2703 so that the filter 2702 is fixed by the filter cover plate 2703. The filter membrane cover plate 2703 is provided with a through hole 2704, so that the material entering the filtering subsystem 05 can pass through the filter membrane cover plate 2703 and then be filtered by passing through the filter membrane 2702. In addition, the diameter of the mesh on the filter membrane 2702 is gradually reduced from the first stage filter chamber 23 to the fourth stage filter chamber 26 so as to realize gradual filtration in the process of extracting the recombinant collagen. Correspondingly, the aperture of the through hole 2704 on the filter membrane cover plate 2703 is also gradually reduced.

Further, the outside of the filter membrane carrier plate 2701 in this embodiment of the application is provided with a handle 28, and the filter membrane carrier plate 2701 can be removed by pulling the handle 28, so that the filter membrane 2702 can be replaced or cleaned conveniently. The lower end of the fourth stage filter chamber 26 is funnel-shaped to facilitate the collection of the filtered material containing recombinant collagen into the receiving subsystem 07.

As shown in fig. 8, the material receiving subsystem 07 provided in the embodiment of the present application includes a material receiving tank 29 and a material receiving pipe 30, where two ends of the material receiving pipe 30 are respectively connected to the material receiving tank 29 and the filtering subsystem 05. The material containing the recombinant collagen after being filtered by the filtering subsystem 05 enters the material receiving tank 29 through the material receiving pipe 30, so that the material collection is realized, and further, the next work is facilitated. Preferably, the receiving tank 29 and the receiving pipe 30 are separable for facilitating the centralized handling of the collected material containing the recombinant collagen.

For extracellular expression recombinant collagen, a feed liquid containing thalli and bacterial liquid is added into a separator 3 through a first liquid inlet 8, a separation rotating motor 2 is started to drive a separation rotating shaft 4 to rotate, and the bacterial liquid of the thalli is separated to form the thalli and the bacterial liquid. Wherein, the thallus gathers in the inside bottom of separator 3, and the fungus liquid gets into between separator 3 and liquid storage tank 1 through the separation hole on separator 3 lateral wall. The driving motor 7 is started to drive the cam 6 to rotate, so that the rubber plug 5 is driven to move up and down. When the rubber stopper 5 moves downward, the rubber stopper 5 is separated from the separator 3, at this time, the bottom of the separator 3 is in an open state, the bacterial cells located at the bottom inside the separator 3 are discharged, and the bacterial liquid is discharged into the first separation subsystem 02 through the liquid outlet. The first rotary motor 11 is started, and the first rotary shaft 14 is rotated by the driving action of the first rotary motor 11, thereby driving the rotary separating blade 15 to rotate. The rotary separation blade 15 stirs the bacterial liquid and the added reagent so that the bacterial liquid and the reagent are uniformly mixed. The materials stirred by the first separation subsystem 02 enter the second stage filtering chamber 24 through the first conveying pipe 04, and the materials containing the recombinant collagen are obtained after the steps of preliminary filtering, impurity removing, multi-step filtering, impurity removing and the like of the multi-stage filtering device 27. And finally is received into the receiving tank 29 through the receiving pipe 30.

For the recombinant collagen expressed in cells, the feed liquid containing thalli and bacterial liquid is added into the separator 3 through the first liquid inlet 8, the separation rotary motor 2 is started to drive the separation rotary shaft 4 to rotate, and the bacterial liquid of the thalli is separated to form the thalli and the bacterial liquid. Wherein, the thallus gathers in the inside bottom of separator 3, and the fungus liquid gets into between separator 3 and liquid storage tank 1 through the separation hole on separator 3 lateral wall. The driving motor 7 is started to drive the cam 6 to rotate, so that the rubber plug 5 is driven to move up and down. When the rubber stopper 5 moves downward, the rubber stopper 5 is separated from the separator 3, and at this time, the bottom of the separator 3 is in an open state, and the cells located at the bottom inside the separator 3 are discharged into the second separation system 03. The second rotating motor 17 is started, and the second rotating shaft 21 rotates under the driving action of the second rotating motor 17, so that the crushing and separating blades 22 are driven to rotate. After the cells enter the second separation tank 16, the lysate is added through the liquid inlet 19. The rotation of the crushing and separating blades 22 agitates and crushes the thalli, so that the recombinant collagen is released into the lysate, and meanwhile, crushed thalli substances are uniformly distributed, thereby preventing blockage in the filtering process. The crushed thallus substances can directly enter the first stage filtering chamber 23 through the second conveying pipe 06, and the materials containing the recombinant collagen are obtained after the steps of preliminary filtering, impurity removing, multi-step filtering, impurity removing and the like of the multi-stage filtering device 27. And finally is received into the receiving tank 29 through the receiving pipe 30.

For the recombinant collagen protein expressed simultaneously in the cell and the cell, the feed liquid containing thalli and bacterial liquid is added into the separator 3 through the first feed liquid port 8, the separation rotary motor 2 is started to drive the separation rotary shaft 4 to rotate, and the bacterial liquid of the thalli is separated to form the thalli and the bacterial liquid. The bacterial liquid is stirred by a first separation subsystem 02 to realize separation and extraction. The stirred bacterial liquid enters the second stage filtering chamber 24 through the first conveying pipe 04, and the materials containing the recombinant collagen are obtained after the steps of preliminary filtering, impurity removing, multi-step filtering, impurity removing and the like of the multi-stage filtering device 27. The thalli are crushed by a second separation system 03 to realize separation and extraction. The crushed thallus substances directly enter the first stage filtering chamber 23 through the second conveying pipe 06, and the materials containing the recombinant collagen are obtained after the steps of preliminary filtering, impurity removing, multi-step filtering, impurity removing and the like of the multi-stage filtering device 27. The material containing recombinant collagen after being filtered by the multi-stage filter 27 is finally received into the receiving tank 29 through the receiving pipe 30.

In the recombinant collagen separation and purification system provided in the embodiment of the present application, different feed liquids respectively enter different separation subsystems according to different expression modes of recombinant collagen, and are collected into the material collecting subsystem 07 after being filtered by the filtering subsystem 05. Therefore, the recombinant collagen separation and purification system provided by the embodiment of the application is added with a bacterial liquid separation subsystem, so that the fermented bacterial liquid containing liquid can be separated into two parts of bacterial liquid and bacterial liquid, and different extraction and purification treatments are respectively carried out on the recombinant collagen in different expression forms according to different added reagents and treatment methods, so that more recombinant collagen can be obtained, and the protein extraction efficiency is improved; meanwhile, the raw material liquid is subjected to multi-layer filtration to ensure the purity of the product. In addition, extracellular expression recombinant collagen does not need thallus fragmentation, so that the extraction process is simplified; the recombinant collagen bacteria which are expressed in the cell and simultaneously expressed in the cell and the cell need to be crushed, and only need 2-3 steps of purification treatment, so that the purification steps are simplified, and the treatment efficiency is improved.

The foregoing description is only of the preferred embodiments of the present application and is not intended to limit the same, but rather, various modifications and variations may be made by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application should be included in the protection scope of the present application.

Claims (10)

1. The recombinant collagen separation and purification system is characterized by comprising a bacterial liquid separation subsystem (01), a first separation subsystem (02) and a second separation subsystem (03), wherein the first separation subsystem (02) and the second separation subsystem (03) are respectively communicated with the bacterial liquid separation subsystem (01), the first separation subsystem (02) is communicated with a filtering subsystem (05) through a first conveying pipe (04), the second separation subsystem (03) is communicated with the filtering subsystem (05) through a second conveying pipe (06), and the bottom of the filtering subsystem (05) is also communicated with a material receiving subsystem (07); the bacterial liquid separation subsystem (01) comprises a liquid storage tank (1), a separation rotating motor (2) arranged on the outer side of the top of the liquid storage tank (1) and a separator (3) arranged in the liquid storage tank (1); a separation rotating shaft (4) is arranged at the top of the inner side of the separator (3), and the separation rotating shaft (4) is connected with the separation rotating motor (2); the device is characterized in that a separation hole is formed in the side wall of the separator (3), a rubber plug (5), a cam (6) and a driving motor (7) are arranged at the bottom of the separator (3), the rubber plug (5) is movably plugged at the bottom of the separator (3), and the cam (6) is respectively connected with the rubber plug (5) and the driving motor (7); the top of liquid storage pot (1) is equipped with first inlet (8) and second inlet (9), first inlet (8) intercommunication the inside of separator (3), second inlet (9) intercommunication the inside of liquid storage pot (1).

2. The recombinant collagen separation and purification system according to claim 1, wherein the first separation subsystem (02) comprises a first separation tank (10), a first rotary motor (11) arranged outside the first separation tank (10), a first separation tank feed inlet (12) arranged at the top of the first separation tank (10), and a first separation tank discharge outlet (13) arranged at the bottom of the first separation tank (10); the first separating tank feed port (12) is communicated with the bacterial liquid separating subsystem (01), and the first separating tank discharge port (13) is communicated with the first conveying pipe (04); the first separating tank (10) is internally provided with a first rotating shaft (14), the first rotating shaft (14) is provided with rotating separating blades (15), and the first rotating shaft (14) is connected with the first rotating motor (11).

3. The recombinant collagen separation and purification system according to claim 1, wherein the second separation system (03) comprises a second separation tank (16), a second rotating electric machine (17) arranged outside the second separation tank (16), a second separation tank feed inlet (18) and a liquid feed inlet (19) arranged at the top of the second separation tank (16), and a second separation tank discharge outlet (20) arranged at the bottom of the second separation tank (16); the second separating tank feed port (18) is communicated with the bacterial liquid separating subsystem (01), and the second separating tank discharge port (20) is communicated with the second conveying pipe (06); the second separation tank (16) is internally provided with a second rotating shaft (21), crushing separation blades (22) are arranged on the second rotating shaft (21), and the second rotating shaft (21) is connected with the second rotating motor (17).

4. The recombinant collagen separation and purification system according to claim 1, wherein the filtering subsystem (05) comprises a first stage filtering chamber (23), a second stage filtering chamber (24), a third stage filtering chamber (25) and a fourth stage filtering chamber (26) which are sequentially arranged from top to bottom, wherein the first stage filtering chamber (23) is communicated with the second conveying pipe (06), the second stage filtering chamber (24) is communicated with the first conveying pipe (04), and the fourth stage filtering chamber (26) is communicated with the material receiving subsystem (07); a filter device (27) is movably arranged between two adjacent filter chambers.

5. The recombinant collagen separation and purification system according to claim 4, wherein the filter device (27) comprises a filter membrane carrier plate (2701), a filter membrane (2702) arranged on the filter membrane carrier plate (2701), and a filter membrane cover plate (2703) covering the filter membrane (2702), and the filter membrane cover plate (2703) is provided with a through hole (2704).

6. The recombinant collagen separation and purification system according to claim 5, wherein the mesh diameter on the filter membrane (2702) is gradually reduced from the first stage filter chamber (23) toward the fourth stage filter chamber (26).

7. The recombinant collagen separation and purification system according to claim 5, wherein a handle (28) is provided on the outside of the filter membrane carrier plate (2701).

8. The recombinant collagen separation and purification system according to claim 1, wherein the material receiving subsystem (07) comprises a material receiving tank (29) and a material receiving pipe (30), and two ends of the material receiving pipe (30) are respectively communicated with the material receiving tank (29) and the filtering subsystem (05).

9. The recombinant collagen separation and purification system according to claim 1, wherein a first delivery pump (08) is provided on the first delivery pipe (04), and a second delivery pump (09) is provided on the second delivery pipe (06).

10. The recombinant collagen separation and purification system according to any one of claims 1 to 9, further comprising a fixed support (010), wherein the bacterial cell liquid separation subsystem (01) and the filtration subsystem (05) are located above the fixed support (010), and wherein the first separation subsystem (02), the second separation subsystem (03) and the material receiving subsystem (07) are located below the fixed support (010).