CN114082321B - Continuous protein renaturation device based on enzyme engineering - Google Patents
Continuous protein renaturation device based on enzyme engineering Download PDFInfo
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- CN114082321B CN114082321B CN202111399159.1A CN202111399159A CN114082321B CN 114082321 B CN114082321 B CN 114082321B CN 202111399159 A CN202111399159 A CN 202111399159A CN 114082321 B CN114082321 B CN 114082321B
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/107—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
- C07K1/113—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides without change of the primary structure
Abstract
The invention relates to the field of protease engineering, in particular to a continuous protein renaturation device based on enzyme engineering, which comprises a first support frame, a second support frame, a reaction frame, a motor, a rotating frame and the like; the first support frame is connected with the second support frame through the mode that the fastener is connected, and fixed mounting has the reaction frame on the first support frame, and second support frame top fixed mounting has the motor, and motor output shaft one end welding has the rotating turret, and the rotating turret is connected with the reaction frame rotation. Through the cooperation of dysmorphism promotion frame and band clamp post rocker, replace artifical manual removal baffle, the continuity adds the mixed solution of protein liquid and dislysate and the protein dislysate of different concentration to the reaction frame in, saves the trouble of manual addition of the mixed solution of protein liquid and dislysate and protein dislysate, realizes full-automatic supply protein liquid and the mixed solution of dislysate and protein dislysate, provides stable liquid serial dilution protein dislysate.
Description
Technical Field
The invention relates to the field of protease engineering, in particular to a continuous protein renaturation device based on enzyme engineering.
Background
The phenomenon that the physical and chemical properties of the protein are changed and the original biological activity is lost due to the fact that the spatial conformation of the molecule is damaged under the influence of certain physical or chemical factors is called denaturation of the protein, when the denaturation condition is not severe and the internal structure of the denatured protein is not greatly changed, the denaturation factor is removed, the denatured protein can restore the natural conformation and the biological activity under the proper condition, the phenomenon is called renaturation of the protein, if the denaturation condition is severe and durable, the denaturation of the protein is irreversible, and if the denaturation condition is not severe, the denaturation is reversible, and the change of the internal structure of the protein molecule is not great.
The protein renaturation method is that the protein dialysate is dialyzed from high to low according to a plurality of gradients, and the method has certain requirements on the solubility of the protein to be renatured, and because a plurality of gradients are needed, the time is needed to calculate manually, the solution is prepared, the dialysate is continuously replaced, the time consumption is long, the concentration of the protein to be dialyzed is low, the probability of protein precipitation is high, the protein precipitation needs to be concentrated at the later stage, and the like, and meanwhile, when the solution is prepared, the solution needs to be stirred manually, but the condition of uneven stirring can occur, and the labor intensity of experimental staff can be high due to long-term manual stirring.
Disclosure of Invention
The invention aims to provide a continuous protein renaturation device based on enzyme engineering, which can automatically supply protein dialysate and exchange liquid, is convenient for realizing protein renaturation, can stir solution and protein dialysate uniformly, and can prevent protein precipitation in the protein renaturation process, so as to solve the problems of time required for manual calculation, solution preparation, continuous exchange of dialysate, long time consumption and uneven stirring of manually stirred solution in the prior art.
The utility model provides a continuous type protein renaturation device based on enzyme engineering, includes first support frame, second support frame, reaction frame, motor, rotating turret, first support frame, fixed bolster, discharge pipe, slip frame, trompil frame, first spring, stirring subassembly and separation subassembly:
the first support frame is connected with the second support frame in a fastening piece connection mode;
the reaction frame is fixedly arranged on the first support frame and is used for providing a reaction place for protein renaturation;
the motor is fixedly arranged at the top of the second support frame and is used for providing driving power for equipment;
the rotating frame is welded at one end of the output shaft of the motor and is rotationally connected with the reaction frame;
the reaction frame is fixedly provided with a first supporting frame;
the fixed support plate is arranged on the first support frame and is used for adding protein solution or protein dialysate;
a discharge pipe fixedly connected to the bottom of the reaction frame for discharging the protein renaturated solution; the discharge pipe is connected with the sliding frame in a sliding manner, and the sliding frame is connected with the reaction frame in a sliding manner;
the perforated frame is fixedly connected with the discharge pipe and is connected with the sliding frame in a sliding manner, and the perforated frame is used for blocking the sliding frame;
the sliding frame is fixedly connected with a first spring, and one end of the first spring is fixedly connected with the discharge pipe;
the stirring assembly is arranged on the rotating frame and is used for uniformly stirring the protein solution and the protein dialysate;
the separation component is arranged on the reaction frame and is used for separating protein solution in the reaction frame at intervals.
Optionally, the stirring assembly comprises a rotating circular plate and a pushing partition plate, the rotating circular plate is connected to the rotating frame in a bolt connection mode, the rotating circular plate is rotationally connected with the reaction frame, the pushing partition plate is connected to the rotating frame in a circumferentially distributed mode, and the pushing partition plate is in contact with the reaction frame.
Optionally, be provided with four on the rotating turret and promote the baffle, promote baffle and the inseparable laminating of reaction frame inner wall, promote the baffle and be used for the protein liquid interval with the different concentration in the reaction frame, promote the baffle and can promote the solution flow in the reaction frame, fully guarantee to add the protein dialysate and take out the solution and do not conflict.
Optionally, the separation subassembly is including separating frame, the second braced frame, the water conservancy diversion frame, the push pedal, the second spring, the check valve, push away the frame, the third support frame, the slip fluting frame and stir the frame, separation frame fixed connection is on the reaction frame, be provided with the second braced frame on the separation frame, second braced frame and reaction frame rigid coupling, be provided with the water conservancy diversion frame in the second braced frame, sliding connection has the push pedal on the second braced frame, fixedly connected with second spring in the push pedal, second spring one end and second braced frame rigid coupling, be provided with the check valve on the push pedal, push away frame fixed connection in the push pedal, reaction frame one side welding has the third support frame, sliding connection has the slip fluting frame on the third support frame, slip fluting frame and push away the spacing cooperation of frame, fixedly connected with stirs the frame on the rotating frame.
Optionally, the injection assembly is fixedly installed on the top surface of the fixed supporting plate, the injection assembly comprises an L-shaped supporting plate, a sliding supporting frame, a liquid storage frame, an input pipe, a first opening frame, a second opening frame, a third opening frame, a fourth supporting frame, a first blocking frame, a second blocking frame, a third spring, a protective folding sleeve, a buoyancy baffle, a baffle plate, a special-shaped pushing frame and a fourth spring, the top surface of the fixed supporting plate is fixedly installed with two L-shaped supporting plates, the sliding supporting frames are connected with the sliding supporting frames on the L-shaped supporting plates, three liquid storage frames are fixedly connected on the two sliding supporting frames together, the input pipe is arranged on the liquid storage frame, a first opening frame is arranged on the other liquid storage frame, a second opening frame is arranged on the other liquid storage frame, a third opening frame is arranged on the other liquid storage frame, a fourth supporting frame is arranged in the liquid storage frame, the first blocking frame is connected with the first blocking frame in a sliding manner, the first blocking frame is in contact with the first opening frame, the other fourth blocking frame is connected with the first opening frame in a sliding manner, the other fourth blocking frame is connected with the third blocking frame in a sliding manner, the second blocking frame is connected with the third opening frame in a sliding manner, the other blocking frame is connected with the third blocking frame in a sliding manner, the third blocking frame is connected with the third blocking frame is in a sliding manner with the third blocking frame is connected with the third blocking frame, the third blocking frame is in a folding frame is in a sliding manner, the third blocking frame is connected with the third blocking frame, the third blocking frame is in a protecting blocking frame is connected with the third blocking frame, the third blocking frame is connected with the third blocking frame, the blocking frame is connected with the third blocking frame. Be provided with the baffle jointly on the three stock solution frames, first trompil frame, second trompil frame and third trompil frame all with baffle rigid coupling, baffle and fixed bolster contact each other, and the dysmorphism promotes the frame sliding connection in fixed bolster, is connected with the fourth spring between fixed bolster and the dysmorphism promotion frame.
Optionally, the automatic ratchet rack comprises a first supporting frame, a second supporting frame, a third supporting frame, a fourth supporting frame, a fifth supporting frame, a sixth spring, a toggle rod, a supporting sliding block, a sliding frame, a pawl, a second torsion spring, a first reset frame, a seventh spring and a second reset frame, wherein the first supporting frame is provided with the switching assembly, the switching assembly comprises a rocker with a clamping column, the fifth spring, the fifth supporting frame is fixedly connected with the fifth supporting frame, the fifth supporting frame is connected with the guide frame in a sliding mode, the sixth spring is connected with the swing plate in a rotating mode, the first torsion spring is connected between the swing plate and the guide frame, the ratchet frame is welded on the sliding supporting frame, the toggle rod is connected with the sliding frame in a uniform arrangement mode, the supporting sliding frame is fixedly connected with the sliding frame on the L-shaped supporting frame, the pawl is connected with the sliding frame in a sliding mode, the pawl is connected with the reset frame in a sliding mode, the first reset frame is connected with the ratchet frame in a sliding mode, and the first reset frame is connected with the ratchet frame.
Optionally, the device further comprises a liquid outlet control assembly, wherein the liquid outlet control assembly is arranged on the sliding frame and comprises a connecting frame, wedge blocks and a pushing rod, the connecting frame is fixedly connected to the sliding frame, the wedge blocks are fixedly connected to the connecting frame, and the pushing rod is fixedly connected to the other sliding support frame.
Optionally, still including preventing depositing the subassembly, prevent depositing the subassembly setting and on the reaction frame, prevent depositing the subassembly including fixed ring gear, stirring frame and spur gear, fixed ring gear fixed mounting is on the reaction frame, rotates to be the mode rotation that four corners distributes on the plectane and is connected with stirring frame, and the rigid coupling has the spur gear on the stirring frame, spur gear and fixed ring gear intermeshing.
The beneficial effects of the invention are as follows:
through the effect of pushing the baffle, promote the baffle with solution interval, the staff adds the mixed solution of protein solution and dislysate and the protein dislysate that different concentration was arranged in proper order according to the gradient from high to low in succession towards the reaction frame, reaches the purpose that can dilute protein dislysate in succession, and in the reaction frame one of them check in the solution can flow to the second braced frame simultaneously, guarantees to add protein dislysate and take out this operation of solution and not conflict, does not need artifical trading liquid, makes the protein renaturation process be in a continuous state, guarantees that protein dislysate concentration presents the linear decline, is convenient for realize the protein renaturation.
Through the cooperation of dysmorphism promotion frame and band clamp post rocker, replace artifical manual removal baffle, the continuity adds the mixed solution of protein liquid and dislysate and the protein dislysate of different concentration to the reaction frame in, saves the trouble of manual addition of the mixed solution of protein liquid and dislysate and protein dislysate, realizes full-automatic supply protein liquid and the mixed solution of dislysate and protein dislysate, provides stable liquid serial dilution protein dislysate.
Through stirring the frame rotation for stirring the frame and stirring the solution in the reaction frame, evenly stir solution and protein dialysate, make solution and protein dialysate intensive mixing, fully dilute protein dialysate, avoid protein precipitation to appear in the protein renaturation in-process simultaneously.
Drawings
Fig. 1 is a schematic perspective view of a first embodiment of the present invention.
Fig. 2 is a schematic perspective view of a second embodiment of the present invention.
Fig. 3 is a schematic perspective view of a first part of the present invention.
Fig. 4 is a schematic perspective view of a second part of the present invention.
Fig. 5 is a schematic perspective view of a third portion of the present invention.
Fig. 6 is a partially disassembled perspective view of the present invention.
Fig. 7 is a schematic perspective view of a first portion of a separator assembly according to the present invention.
FIG. 8 is a schematic perspective view, partially in section, of a separator assembly of the present invention.
Fig. 9 is a schematic perspective view of a second portion of the separation assembly of the present invention.
Fig. 10 is a schematic view of a partial perspective view of an injection assembly according to the present invention.
Fig. 11 is a schematic perspective view, partially in section, of a first type of injection assembly of the present invention.
Fig. 12 is a schematic view of a second partially cut-away perspective structure of an infusion assembly of the present invention.
Fig. 13 is a schematic perspective view, partially in section, of a third type of infusion assembly in accordance with the present invention.
Fig. 14 is a schematic perspective view of a first portion of a switching assembly according to the present invention.
Fig. 15 is a schematic view of a second partial perspective view of the switching assembly of the present invention.
Fig. 16 is a schematic view of a third part of a third perspective structure of the switching assembly of the present invention.
Fig. 17 is a schematic view of a third partial perspective view of the switching assembly of the present invention.
FIG. 18 is a schematic view showing a partial perspective structure of the liquid outlet control assembly of the present invention.
Fig. 19 is a schematic perspective view of a sliding support and a push rod according to the present invention.
FIG. 20 is a schematic perspective view of an anti-settling assembly according to the present invention.
FIG. 21 is a schematic view showing a partial perspective structure of an anti-settling assembly according to the present invention.
The reference symbols in the drawings: 1: first support frame, 21: second support frame, 22: reaction frame, 23: motor, 24: rotating frame, 25: first support frame, 26: fixed support plate, 27: a discharge pipe, 28: slide frame, 29: perforated rack, 210: first spring, 3: stirring assembly, 31: rotating the circular plate, 32: pushing the partition plate, 4: separation assembly, 41: separation frame, 42: second support frame, 421: flow guiding frame, 43: push plate, 44: second spring, 45: check valve, 46: pushing frame, 47: third support frame, 48: slide slotted frame, 49: toggle rack, 5: injection assembly, 51: l-shaped support plate, 52: sliding support, 53: liquid storage frame, 531: input tube, 532: first aperture frame, 533: second aperture frame, 534: third aperture frame, 54: fourth support frame, 55: first blockage rack 551: second plug rack, 552: third plug rack, 56: third spring, 57: protective fold sleeve, 58: buoyancy stopper 581: baffle plate, 59: dysmorphism promotes frame, 510: fourth spring, 6: switching component, 61: with clamping column rocker 6101: fifth spring, 62: fifth support frame, 63: guide rack 631: swing plate, 632: first torsion spring, 64: sixth spring, 65: ratchet bar, 66: toggle rod, 67: support slide, 68: carriage, 69: pawl, 610: second torsion spring, 611: first reset frame, 612: seventh spring, 613: second frame that resets, 7: liquid outlet control assembly, 71: connecting frame, 72: wedge block, 73: push rod, 8: anti-settling assembly, 81: stationary ring gear, 82: agitation frame, 83: spur gears.
Detailed Description
In the description of the present invention, unless otherwise indicated, the meaning of "a plurality" is two or more; the terms "upper," "lower," "left," "right," "inner," "outer," "front," "rear," "head," "tail," and the like are used as an orientation or positional relationship based on that shown in the drawings, merely to facilitate description of the invention and to simplify the description, and do not indicate or imply that the devices or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore should not be construed as limiting the invention. Furthermore, the terms "first," "second," "third," and the like are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
Example 1
The continuous protein renaturation device based on enzyme engineering comprises a first support frame 1, a second support frame 21, a reaction frame 22, a motor 23, a rotating frame 24, a first support frame 25, a fixed support plate 26, a discharge pipe 27, a sliding frame 28, an opening frame 29, a first spring 210, an agitating component 3 and a separating component 4, wherein the first support frame 1 is connected with the second support frame 21 through a fastening piece, the first support frame 1 is fixedly provided with the reaction frame 22, the top of the second support frame 21 is fixedly provided with a motor 23 for driving, one end of an output shaft of the motor 23 is welded with the rotating frame 24, the rotating frame 24 is rotationally connected with the reaction frame 22, the first support frame 25 is fixedly provided with the fixed support plate 26, the discharge pipe 27 is fixedly connected with the bottom of the reaction frame 22, the discharge pipe 27 is used for discharging solution, the sliding frame 28 is connected with the sliding frame 28, the opening frame 29 is fixedly connected with the sliding frame 27, the opening frame 29 is fixedly connected with the sliding frame 28, the opening frame 28 is used for fixedly connecting the sliding frame 28 with the sliding frame 22 with the first spring 22, the agitating component 3 is fixedly arranged on the first support frame 22, and the agitating component is fixedly connected with the first spring 2, and the agitating component is fixedly arranged on the first support frame 22.
The stirring assembly 3 comprises a rotary circular plate 31 and a pushing partition plate 32, the rotary circular plate 31 is connected to the rotary frame 24 in a bolt connection mode, the rotary circular plate 31 is connected with the reaction frame 22 in a rotary mode, the pushing partition plate 32 is connected to the rotary frame 24 in a circumferentially distributed mode, the pushing partition plate 32 is in contact with the reaction frame 22, and the pushing partition plate 32 is used for spacing the solution in the reaction frame 22.
The separation assembly 4 comprises a separation frame 41, a second support frame 42, a flow guiding frame 421, a push plate 43, a second spring 44, a one-way valve 45, a push frame 46, a third support frame 47, a sliding slotted frame 48 and a toggle frame 49, wherein the separation frame 41 is fixedly connected to the reaction frame 22, the second support frame 42 is arranged on the separation frame 41, the second support frame 42 is fixedly connected with the reaction frame 22, the flow guiding frame 421 is arranged in the second support frame 42, the second support frame 42 is connected with the push plate 43 in a sliding manner, the push plate 43 is used for extruding a solution in the second support frame 42, the push plate 43 is fixedly connected with the second spring 44, one end of the second spring 44 far away from the push plate 43 is fixedly connected with the second support frame 42, the push plate 43 is provided with the one-way valve 45, the push frame 46 is fixedly connected with the push plate 43, one side of the reaction frame 22 is welded with the third support frame 47, the third support frame 47 is connected with the sliding slotted frame 48 in a sliding manner, the sliding slotted frame 48 is in a limiting fit with the push frame 46, the toggle frame 49 is fixedly connected with the rotary frame 24, and the toggle frame 49 is used for pushing the sliding slotted frame 48 to move downwards.
When protein renaturation is required to be realized, a worker manually starts the motor 23, the prepared protein solution is added into the reaction frame 22 through the liquid injection port on the fixed support plate 26, the separation plate 32 is pushed to separate the solution, the output shaft of the motor 23 rotates to drive the rotating frame 24 and the device on the rotating frame to rotate clockwise, the separation plate 32 is pushed to push the solution in the reaction frame 22 to flow, then the protein dialysate with higher concentration is added into the reaction frame 22, one lattice of solution can be mixed with the protein dialysate, and the solution can dilute the protein dialysate. Simultaneously, the rotating frame 24 rotates to drive the stirring frame 49 to rotate, the stirring frame 49 pushes the sliding slotting frame 48 to move downwards, and the sliding slotting frame 48 pushes the pushing frame 46 and the device thereon to move downwards, so that part of the solution in one of the cells in the reaction frame 22 flows into the second support frame 42. When the toggle frame 49 is separated from the sliding slotted frame 48, the compressed second spring 44 resets to drive the push plate 43 to move upwards, the push plate 43 extrudes the solution in the second supporting frame 42, so that the solution in the second supporting frame 42 flows out through the one-way valve 45 and the flow guiding frame 421, a worker collects the solutions with different concentrations, after pushing the partition plate 32 to rotate for one circle, the worker adds protein dialysate with lower concentration into the reaction frame 22, and repeats the above operation, and the worker continuously adds protein dialysate with different concentrations into the reaction frame 22 from high to low according to the gradient, so as to continuously dilute the protein dialysate, and simultaneously ensure that the operation of adding the protein dialysate and extracting the solution is not in conflict, so that the protein renaturation process is in a continuous state, and the concentration of the protein dialysate is ensured to be linearly decreased.
Example 2
On the basis of the embodiment 1, as shown in fig. 10-14, the device further comprises an injection assembly 5, the top surface of the fixed support plate 26 is fixedly provided with the injection assembly 5, the injection assembly 5 is used for adding protein dialysate into the reaction frame 22, the injection assembly 5 comprises an L-shaped support plate 51, a sliding support frame 52, a liquid storage frame 53, an input pipe 531, a first opening frame 532, a second opening frame 533, a third opening frame 534, a fourth support frame 54, a first blocking frame 55, a second blocking frame 551, a third blocking frame 552, a third spring 56, a protective folding sleeve 57, a buoyancy blocking frame 58, a blocking plate 581, a special-shaped pushing frame 59 and a fourth spring 510, the top surface of the fixed support plate 26 is fixedly provided with two L-shaped support plates 51, the sliding support frames 52 are connected to the L-shaped support plates 51 in a sliding manner, three liquid storage frames 53 are fixedly connected to the two sliding support frames 52 together, the liquid storage frames 53 are provided with the input pipe 531, wherein one liquid storage frame 53 is provided with a first open hole frame 532, the other liquid storage frame 53 far away from the first open hole frame 532 is provided with a second open hole frame 533, the other liquid storage frame 53 is provided with a third open hole frame 534, a fourth support frame 54 is arranged in the liquid storage frame 53, wherein a first blocking frame 55 is connected on the fourth support frame 54 in a sliding manner, the first blocking frame 55 is contacted with the first open hole frame 532 mutually, the first blocking frame 55 is used for blocking a liquid outlet hole on the first open hole frame 532, the other fourth support frame 54 is connected with a second blocking frame 551 in a sliding manner, the second blocking frame 551 is contacted with the second open hole frame 533 mutually, the second blocking frame 551 is used for blocking the liquid outlet hole on the second open hole frame 533, the other fourth support frame 54 is connected with a third blocking frame 552 in a sliding manner, the third blocking frame 552 is contacted with the third open hole frame 534 mutually, the third blocking frame 552 is used for blocking the liquid outlet hole on the third open hole frame 534, the third spring 56 is connected between the first blocking frame 55 and one fourth supporting frame 54, the third spring 56 is connected between the second blocking frame 551 and one fourth supporting frame 54, the third blocking frame 552 is connected with the other fourth supporting frame 54, the third spring 56 is connected between the third blocking frame 552 and the other fourth supporting frame 54, the protective folding sleeve 57 is fixedly connected to the fourth supporting frame 54, the protective folding sleeve 57 is fixedly connected with the first opening frame 532, the protective folding sleeve 57 is fixedly connected with the second opening frame 533, the protective folding sleeve 57 is fixedly connected with the third opening frame 534, the buoyancy blocking frame 58 is slidingly connected to the liquid storage frame 53, the blocking plate 581 is jointly arranged on the three liquid storage frames 53, the first opening frame 532, the second opening frame 533 and the third opening frame 534 are fixedly connected with the blocking plate 581, the blocking plate 581 is mutually contacted with the fixed supporting plate 26, the special-shaped pushing frame 59 is slidingly connected to the fixed supporting plate 26, the special-shaped pushing frame 59 is used for pushing the second blocking frame 551, the third blocking frame 552 or the first blocking frame 55 to move upwards, and the fourth spring 510 is connected between the fixed supporting plate 26 and the special-shaped pushing frame 59.
The staff adds the mixed solution of protein liquid and dialysate and the protein dialysate that different concentrations are arranged in proper order according to the gradient from high to low into stock solution frame 53 through input tube 531, and the liquid level in stock solution frame 53 can rise thereupon, and the mixed solution of protein liquid and dialysate produces buoyancy and promotes buoyancy and keeps off frame 58 upward movement, and buoyancy keeps off frame 58 gradually blocks the inlet on the stock solution frame 53 for the inlet velocity reduces according to the liquid level rising gradually. The worker then manually pushes the barrier plate 581 toward the direction approaching the profiled pushing frame 59 so that the second open-hole frame 533 corresponds to the liquid filling port on the fixed support plate 26. The stirring frame 49 rotates to push the special-shaped pushing frame 59 to move upwards, the special-shaped pushing frame 59 can push the second blocking frame 551 to move upwards, and the second blocking frame 551 does not block the liquid outlet hole on the second open hole frame 533, so that the mixed liquid of the protein liquid and the dialysis liquid in the liquid storage frame 53 flows into the reaction frame 22. When the toggle frame 49 is separated from the special-shaped pushing frame 59, the compressed fourth spring 510 resets to drive the special-shaped pushing frame 59 to move downwards to reset, and then the compressed third spring 56 resets to drive the second blocking frame 551 to move downwards to reset, so that the second blocking frame 551 blocks the liquid outlet on the second open hole frame 533. The above operation was repeated, and a mixed solution of the protein solution and the dialysate was added to each cell of the reaction frame 22.
After the toggle frame 49 rotates one turn, the worker pushes the blocking plate 581 again toward the direction approaching the special-shaped pushing frame 59, so that the third opening frame 534 corresponds to the liquid injection port on the fixed support plate 26. The upward movement of the special-shaped pushing frame 59 pushes the third blocking frame 552 to move upward, and the third blocking frame 552 does not block the liquid outlet hole on the third opening frame 534, so that the protein dialysate with higher concentration in the liquid storage frame 53 flows into the reaction frame 22. When the special-shaped pushing frame 59 moves downwards to reset, the compressed third spring 56 resets to drive the third blocking frame 552 to move downwards to reset, so that the third blocking frame 552 blocks the liquid outlet hole on the third opening frame 534.
After the toggle frame 49 rotates for one turn, the worker pushes the blocking plate 581 towards the direction approaching to the special-shaped pushing frame 59 again, so that the first perforated frame 532 corresponds to the liquid injection port on the fixed support plate 26. The upward movement of the special-shaped pushing frame 59 pushes the first blocking frame 55 to move upward, and the first blocking frame 55 does not block the liquid outlet hole on the first opening frame 532, so that the protein dialysate with lower concentration in the liquid storage frame 53 flows into the reaction frame 22. When the special-shaped pushing frame 59 moves downwards to reset, the compressed third spring 56 resets to drive the first blocking frame 55 to move downwards to reset, so that the first blocking frame 55 blocks the liquid outlet hole on the first open hole frame 532. The above operation is repeated, and the mixed solution of the protein solution and the dialyzate and the protein dialyzate with different concentrations are continuously added into the reaction frame 22, so that the trouble of manually adding the mixed solution of the protein solution and the dialyzate and the protein dialyzate is omitted, and the stable liquid is further provided for continuously diluting the protein dialyzate.
Example 3
Based on embodiment 2, as shown in fig. 14-17, the device further comprises a switching component 6, a switching component 6 is arranged on the first supporting frame 25, the switching component 6 is used for switching the concentration of the added protein dialysate, the switching component 6 comprises a rocker 61 with a clamping column, a fifth spring 6101, a fifth supporting frame 62, a guide frame 63, a swing plate 631, a first torsion spring 632, a sixth spring 64, a ratchet rack 65, a toggle rod 66, a supporting slide block 67, a sliding frame 68, a pawl 69, a second torsion spring 610, a first reset frame 611, a seventh spring 612 and a second reset frame 613, the rocker 61 with the clamping column is fixedly connected on the rotating frame 24, a fifth spring 6101 is connected between the L-shaped supporting plate 51 and the sliding supporting frame 52, a fifth supporting frame 62 is fixedly connected on the first supporting frame 25, a guide frame 63 is slidingly connected on the fifth supporting frame 62, the fifth support frame 62 is connected with a sixth spring 64, one end of the sixth spring 64 far away from the fifth support frame 62 is connected with a guide frame 63, a swing plate 631 is rotatably connected on the guide frame 63 close to the fifth support frame 62, a first torsion spring 632 is connected between the swing plate 631 and the guide frame 63, a ratchet bar 65 is welded on a sliding support frame 52, a toggle rod 66 is connected on the sliding support frame 52 in a uniformly arranged mode, a support slide block 67 is fixedly connected on the L-shaped support plate 51, a sliding frame 68 is slidingly connected on the support slide block 67, a pawl 69 is rotatably connected on the sliding frame 68, the pawl 69 is mutually contacted with the ratchet bar 65, the pawl 69 is used for clamping the ratchet bar 65, a second torsion spring 610 is connected between the pawl 69 and the sliding frame 68, a first reset frame 611 is slidingly connected on the ratchet bar 65, the first reset frame 611 is mutually contacted with the sliding frame 68, the first reset frame 611 is used for pushing the sliding frame 68 and the device thereon to move towards the direction approaching the sliding support frame 52, a seventh spring 612 is connected between the first reset frame 611 and the ratchet bar 65, and a second reset frame 613 is fixedly connected to the ratchet bar 65 far from the first reset frame 611.
The rotating frame 24 rotates to push the rocker 61 with the clamping column to rotate clockwise, the rocker 61 with the clamping column pushes the guide frame 63 and the upper device thereof to move upwards, the swinging plate 631 pushes the toggle rod 66 and the upper device thereof to move towards the direction close to the special-shaped pushing frame 59, the blocking plate 581 is moved towards the direction close to the special-shaped pushing frame 59 instead of manual operation, the ratchet rack 65 pushes the pawl 69 to swing, and then the pawl 69 clamps the ratchet rack 65 under the action of the second torsion spring 610, so that the fifth spring 6101 is prevented from resetting to drive the sliding support frame 52 and the upper device thereof to move towards the direction far away from the special-shaped pushing frame 59.
When the rotating frame 24 is separated from the rocker 61 with a clamping column, the compressed sixth spring 64 resets to drive the guide frame 63 and the device thereon to move downwards, the toggle rod 66 pushes the swing plate 631 to swing, and the compressed first torsion spring 632 resets to drive the swing plate 631 to swing and reset. When the second reset frame 613 contacts with the sliding frame 68, the second reset frame 613 pushes the sliding frame 68 and the upper device thereof to move away from the sliding support frame 52, so that the pawl 69 does not clamp the ratchet bar 65 any more, the stretched fifth spring 6101 resets to drive the sliding support frame 52 and the upper device thereof to move away from the profiled pushing frame 59 for resetting, and then the first reset frame 611 pushes the sliding frame 68 and the upper device thereof to move toward the direction approaching the sliding support frame 52, and the pawl 69 clamps the ratchet bar 65 again.
Example 4
18-19, further include a liquid outlet control assembly 7, where the liquid outlet control assembly 7 is disposed on the sliding frame 28, the liquid outlet control assembly 7 is used to control the discharge tube 27 to discharge the solution, the liquid outlet control assembly 7 includes a connecting frame 71, a wedge block 72 and a push rod 73, the connecting frame 71 is fixedly connected to the sliding frame 28, the wedge block 72 is fixedly connected to the connecting frame 71 far from the sliding frame 28, and the push rod 73 is fixedly connected to the other sliding support 52.
When the sliding support frame 52 and the device thereon move in the direction approaching to the special-shaped pushing frame 59, the pushing rod 73 will contact with the wedge block 72, the pushing rod 73 will push the wedge block 72 and the connecting frame 71 to move downward, the connecting frame 71 will drive the sliding frame 28 to move downward, so that the opening frame 29 will not block the sliding frame 28 any more, and the solution in the reaction frame 22 will be discharged from the discharge tube 27 through the sliding frame 28. The solution in the reaction frame 22 is pushed to flow by pushing the partition plate 32 so that the solution in the reaction frame 22 is entirely discharged from the discharge pipe 27. When the sliding support frame 52 and the device thereon move away from the special-shaped pushing frame 59, the pushing rod 73 is separated from the wedge block 72, and the compressed first spring 210 resets to drive the sliding frame 28 to move upwards for resetting, and the opening frame 29 plugs the sliding frame 28.
Example 5
On the basis of embodiment 4, as shown in fig. 20-21, the device further comprises an anti-precipitation component 8, the anti-precipitation component 8 is arranged on the reaction frame 22, the anti-precipitation component 8 is used for avoiding protein precipitation in the protein renaturation process, the anti-precipitation component 8 comprises a fixed gear ring 81, an stirring frame 82 and a spur gear 83, the fixed gear ring 81 is fixedly arranged on the reaction frame 22, the stirring frame 82 is rotationally connected with the stirring frame 82 in a manner of being distributed in four corners on the rotary circular plate 31, the stirring frame 82 is used for stirring the solution in the reaction frame 22, the spur gear 83 is fixedly connected on the stirring frame 82, and the spur gear 83 is mutually meshed with the fixed gear ring 81.
The rotating frame 24 can drive the rotating circular plate 31 and the upper device thereof to rotate, and the spur gear 83 and the stirring frame 82 rotate under the action of the fixed gear ring 81, so that the stirring frame 82 stirs the solution in the reaction frame 22, the solution is fully mixed with the protein dialysate, the protein dialysate is fully diluted, and protein precipitation is avoided in the protein renaturation process.
The embodiments of the invention have been presented for purposes of illustration and description, and are not intended to be exhaustive or limited to the invention in the form disclosed. Many modifications and variations will be apparent to those of ordinary skill in the art. The embodiments were chosen and described in order to best explain the principles of the invention and the practical application, and to enable others of ordinary skill in the art to understand the invention for various embodiments with various modifications as are suited to the particular use contemplated.
Claims (5)
1. Continuous type protein renaturation device based on enzyme engineering, its characterized in that, including first support frame, second support frame, reaction frame, motor, rotating turret, first support frame, fixed bolster, discharge pipe, slip frame, trompil frame, first spring, stirring subassembly and separation subassembly:
the first support frame is connected with the second support frame in a fastening piece connection mode;
the reaction frame is fixedly arranged on the first support frame and is used for providing a reaction place for protein renaturation;
the motor is fixedly arranged at the top of the second support frame and is used for providing driving power for equipment;
the rotating frame is welded at one end of the output shaft of the motor and is rotationally connected with the reaction frame;
the reaction frame is fixedly provided with a first supporting frame;
the fixed support plate is arranged on the first support frame and is used for adding protein solution or protein dialysate;
a discharge pipe fixedly connected to the bottom of the reaction frame for discharging the protein renaturated solution;
the discharge pipe is connected with the sliding frame in a sliding manner, and the sliding frame is connected with the reaction frame in a sliding manner;
the perforated frame is fixedly connected with the discharge pipe and is connected with the sliding frame in a sliding manner, and the perforated frame is used for blocking the sliding frame;
the sliding frame is fixedly connected with a first spring, and one end of the first spring is fixedly connected with the discharge pipe;
the stirring assembly is arranged on the rotating frame and is used for uniformly stirring the protein solution and the protein dialysate;
the separation component is arranged on the reaction frame and is used for separating protein solution in the reaction frame at intervals;
the stirring assembly comprises a rotating circular plate and a pushing partition plate, the rotating circular plate is connected to the rotating frame in a bolt connection mode, the rotating circular plate is connected with the reaction frame in a rotating mode, the pushing partition plate is connected to the rotating frame in a circumferential distribution mode, and the pushing partition plate is in contact with the reaction frame;
four pushing separation plates are arranged on the rotating frame and are closely attached to the inner wall of the reaction frame, the pushing separation plates are used for separating protein liquids with different concentrations in the reaction frame, the pushing separation plates can push the solution in the reaction frame to flow, and the situation that the adding of protein dialysate and the extraction of the solution do not conflict is fully guaranteed;
the separation assembly comprises a separation frame, a second supporting frame, a flow guiding frame, a push plate, a second spring, a one-way valve, a pushing frame, a third supporting frame, a sliding slotting frame and a pushing frame, wherein the separation frame is fixedly connected to a reaction frame, the second supporting frame is arranged on the separation frame and fixedly connected with the reaction frame, the flow guiding frame is arranged in the second supporting frame, the push plate is connected to the second supporting frame in a sliding mode, the second spring is fixedly connected to the push plate, one end of the second spring is fixedly connected with the second supporting frame, the one-way valve is arranged on the push plate, the pushing frame is fixedly connected to the push plate, the third supporting frame is welded to one side of the reaction frame, the sliding slotting frame is connected to the sliding slotting frame in a limiting mode, and the pushing frame is fixedly connected to the rotating frame.
2. The continuous protein renaturation device based on enzyme engineering according to claim 1, further comprising an injection component, wherein the injection component is fixedly arranged on the top surface of the fixed support plate, the injection component comprises an L-shaped support plate, a sliding support frame, a liquid storage frame, an input pipe, a first opening frame, a second opening frame, a third opening frame, a fourth support frame, a first blocking frame, a second blocking frame, a third spring, a protective folding sleeve, a buoyancy baffle frame, a baffle plate, a special-shaped pushing frame and a fourth spring, two L-shaped support plates are fixedly arranged on the top surface of the fixed support plate, the sliding support frames are connected on the L-shaped support plate, a third liquid storage frame is fixedly connected on the two sliding support frames, an input pipe is arranged on the liquid storage frame, a first opening frame is arranged on the first liquid storage frame, a second opening frame is arranged on the other liquid storage frame, a fourth support frame is arranged in the frame, the first blocking frame is connected with the first blocking frame in a sliding manner, the first blocking frame is contacted with the first opening frame, the other fourth supporting frame is connected with the first opening frame, the second blocking frame is connected with the fourth opening frame in a sliding manner, the other protective folding frame is fixedly connected with the third opening frame, the other protective folding frame is fixedly connected with the third opening frame, the third opening frame is connected with the third opening frame is fixedly connected with the third opening frame, and the third opening frame is fixedly connected with the third opening frame, the other part is protected the folding cover and is fixedly connected with third trompil frame, and sliding connection has buoyancy to keep off the frame on the stock solution frame, is provided with the baffle jointly on the three stock solution frames, and first trompil frame, second trompil frame and third trompil frame all are fixedly connected with the baffle, and the baffle contacts each other with fixed support plate, and the dysmorphism promotes the frame sliding connection in fixed support plate, is connected with the fourth spring between fixed support plate and the dysmorphism promotion frame.
3. The continuous protein renaturation device based on enzyme engineering is characterized by further comprising a switching component, wherein the switching component is arranged on the first supporting frame and comprises a rocker with a clamping column, a fifth spring, a fifth supporting frame, a guide frame, a swinging plate, a first torsion spring, a sixth spring, a ratchet rack, a poking rod, a supporting sliding block, a sliding frame, a pawl, a second torsion spring, a first reset frame, a seventh spring and a second reset frame, the rocker with the clamping column is fixedly connected on the rotating frame, a fifth spring is connected between the L-shaped supporting plate and the sliding supporting frame, the fifth supporting frame is fixedly connected with the fifth supporting frame, a guide frame is connected on the fifth supporting frame in a sliding mode, a sixth spring is connected with the guide frame at one end of the fifth spring, the swinging plate is connected with the guide frame in a rotating mode, the first torsion spring is connected between the swinging plate and the guide frame, the ratchet rack is welded on a sliding frame, the poking rod is connected on the same sliding supporting frame in a uniform arrangement mode, the supporting sliding block is fixedly connected on the L-shaped sliding block, the supporting frame is connected with the sliding frame, the ratchet rack is connected with the reset frame, the ratchet rack is connected with the second ratchet rack in a sliding mode, and the ratchet rack is fixedly connected with the second ratchet rack is connected with the reset frame.
4. The continuous protein renaturation device based on enzyme engineering is characterized by further comprising a liquid outlet control component, wherein the liquid outlet control component is arranged on the sliding frame and comprises a connecting frame, a wedge-shaped block and a pushing rod, the connecting frame is fixedly connected to the sliding frame, the wedge-shaped block is fixedly connected to the connecting frame, and the pushing rod is fixedly connected to the other sliding support frame.
5. The continuous protein renaturation device based on enzyme engineering is characterized by further comprising an anti-sedimentation component, wherein the anti-sedimentation component is arranged on the reaction frame and comprises a fixed gear ring, an agitating frame and a spur gear, the fixed gear ring is fixedly arranged on the reaction frame, the agitating frame is rotationally connected to the rotating circular plate in a manner of being distributed in four corners, the spur gear is fixedly connected to the agitating frame, and the spur gear is meshed with the fixed gear ring.
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| EP1845103B1 (en) * | 2006-04-10 | 2015-02-25 | Boehringer Ingelheim RCV GmbH & Co KG | Method for refolding a protein |
| CN107857794B (en) * | 2017-12-20 | 2024-04-02 | 天康生物制药有限公司 | Protein dialysis device and system |
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| CN113266548B (en) * | 2021-07-19 | 2021-10-08 | 山东高原油气装备有限公司 | Hydraulic bidirectional slurry pump for preventing blockage in oil exploitation |
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| CN111494690A (en) * | 2020-05-07 | 2020-08-07 | 廖曦 | Quick blow-off device of medical gauze mask production ethylene oxide |
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