CN114082321A - Continuous type protein renaturation device based on enzyme engineering - Google Patents
Continuous type protein renaturation device based on enzyme engineering Download PDFInfo
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- CN114082321A CN114082321A CN202111399159.1A CN202111399159A CN114082321A CN 114082321 A CN114082321 A CN 114082321A CN 202111399159 A CN202111399159 A CN 202111399159A CN 114082321 A CN114082321 A CN 114082321A
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- 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
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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 a second support frame in a manner of being connected through a fastener, the first support frame is fixedly provided with a reaction frame, the top of the second support frame is fixedly provided with a motor, one end of an output shaft of the motor is welded with a rotating frame, and the rotating frame is rotatably connected with the reaction frame. Through the cooperation of dysmorphism propelling movement frame and area card post rocker, replace artifical manual removal baffler, the mixed liquid of protein liquid and dislysate and the protein dislysate of different concentrations of continuity add to the reaction frame in, save the trouble of artifical manual mixed liquid and the protein dislysate that adds protein liquid and dislysate, realize the mixed liquid and the protein dislysate of full-automatic supply protein liquid and dislysate, provide stable liquid and dilute the protein dislysate continuously.
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 spatial conformation of a molecule is destroyed because of the influence of certain physical or chemical factors, so that the physicochemical property of the protein is changed and the original biological activity is lost is called protein denaturation, when the denaturation condition is not severe and the internal structure of the denatured protein is not greatly changed, the denaturation factor is removed, and the denatured protein can restore the natural conformation and the biological activity under the proper condition, the phenomenon is called protein renaturation, if the denaturation condition is severe and durable, the protein denaturation 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 large.
The protein renaturation method is characterized in that protein dialysate is dialyzed from high to low according to a plurality of gradients, the method has certain requirements on the solubility of protein to be renatured, the multiple gradients are needed, therefore, the time is needed to be calculated 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 protein precipitation probability is high, the concentration and other treatment are needed in the later period, meanwhile, when the solution is prepared, the solution needs to be stirred manually, the condition of uneven stirring can occur, and the labor intensity of experimenters 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 and replace protein dialysate, is convenient to realize protein renaturation, can uniformly stir the solution and the protein dialysate, and can reduce protein precipitation in the protein renaturation process, so as to solve the problems of long time consumption and uneven stirring caused by manual stirring of the solution due to manual calculation time, solution preparation and continuous dialysate replacement in the background technology.
A continuous protein renaturation device based on enzyme engineering, including first support frame, second support frame, reaction frame, motor, rotating turret, first support frame, fixed support plate, discharge pipe, slip frame, trompil frame, first spring, stirring subassembly and separator assembly:
the first support frame is connected with the second support frame in a fastener connection mode;
the reaction frame is fixedly arranged on the first support frame and used for providing a reaction site for protein renaturation;
the motor is fixedly arranged at the top of the second support frame and used for providing driving power for equipment;
one end of an output shaft of the motor is welded with the rotating frame, and the rotating frame is rotatably connected with the reaction frame;
the reaction frame is fixedly provided with a first support frame;
the first support frame is provided with a fixed support plate, and the fixed support plate is used for adding protein solution or protein dialysate;
the discharge pipe is fixedly connected to the bottom of the reaction frame and is used for discharging the protein renaturation 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 discharge pipe is fixedly connected with the tapping frame, the tapping frame is connected with the sliding frame in a sliding mode, and the tapping frame is used for plugging 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 component 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 the 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 rotatably connected with the reaction frame, 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.
Optionally, four pushing separation plates are arranged on the rotating frame, the pushing separation plates are tightly 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 it is fully guaranteed that the protein dialysate is added and the solution is not drawn out.
Optionally, the separation assembly comprises a separation frame, a second support frame, a flow guide frame, a push plate, a second spring, a check valve, a push frame, a third support frame, a sliding slotting frame and a poking frame, the separation frame is fixedly connected to the reaction frame, the second support frame is arranged on the separation frame and fixedly connected to the reaction frame, the flow guide frame is arranged in the second support frame, the push plate is slidably connected to the second support frame, the second spring is fixedly connected to the push plate, one end of the second spring is fixedly connected to the second support frame, the check valve is arranged on the push plate, the push frame is fixedly connected to the push plate, the third support frame is welded to one side of the reaction frame, the sliding slotting frame is slidably connected to the third support frame, the sliding slotting frame is in limit fit with the push frame, and the poking frame is fixedly connected to the rotating frame.
Optionally, the device further comprises an injection assembly, the injection assembly is fixedly installed on the top surface of the fixed support plate, the injection assembly comprises an L-shaped support plate, a sliding support frame, a liquid storage frame, an input tube, a first open hole frame, a second open hole frame, a third open hole frame, a fourth support frame, a first blocking frame, a second blocking frame, a third spring, a protective folding sleeve, a buoyancy blocking frame, a blocking plate, a special-shaped pushing frame and a fourth spring, the two L-shaped support plates are fixedly installed on the top surface of the fixed support plate, the sliding support frames are slidably connected on the L-shaped support plates, the three liquid storage frames are fixedly connected on the two sliding support frames together, the input tube is arranged on the liquid storage frame, the first open hole frame is arranged on one liquid storage frame, the second open hole frame is arranged on the other liquid storage frame, the third open hole frame is arranged on the other liquid storage frame, the fourth support frame is arranged in the liquid storage frame, the first blocking frame is slidably connected on the fourth support frame, the first blocking frame is in contact with the first hole opening frame, the other fourth supporting frame is in sliding connection with a second blocking frame, the second blocking frame is in contact with the second hole opening frame, the other fourth supporting frame is in sliding connection with a third blocking frame, the third blocking frame is in contact with the third hole opening frame, a third spring is connected between the first blocking frame and one fourth supporting frame, a third spring is connected between the second blocking frame and one fourth supporting frame, a third spring is connected between the third blocking frame and the other fourth supporting frame, a protective folding sleeve is fixedly connected onto the fourth supporting frame, one protective folding sleeve is fixedly connected onto the first hole opening frame, the other protective folding sleeve is fixedly connected onto the second hole opening frame, the other protective folding sleeve is fixedly connected onto the third hole opening frame, a buoyancy blocking frame is in sliding connection onto the liquid storage frame, blocking plates are jointly arranged on the three liquid storage frames, and the first hole opening frame is fixedly connected onto the second hole opening frame, The second open-hole frame and the third open-hole frame are fixedly connected with the blocking plate, the blocking plate is in contact with the fixed supporting plate, the special-shaped pushing frame is connected to the fixed supporting plate in a sliding mode, and a fourth spring is connected between the fixed supporting plate and the special-shaped pushing frame.
Optionally, the device further comprises a switching assembly, the first support frame is provided with the switching assembly, the switching assembly comprises a rocker with a clamping column, a fifth spring, a fifth support frame, a guide frame, a swing plate, a first torsion spring, a sixth spring, a ratchet bar, a poke rod, a support slider, a sliding frame, a pawl, a second torsion spring, a first reset frame, a seventh spring and a second reset frame, the rotating frame is fixedly connected with the rocker with the clamping column, the fifth spring is connected between the L-shaped support plate and the sliding support frame, the first support frame is fixedly connected with the fifth support frame, the fifth support frame is slidably connected with the guide frame, the fifth support frame is connected with the sixth spring, one end of the sixth spring is connected with the guide frame, the guide frame is rotatably connected with the swing plate, the first torsion spring is connected between the swing plate and the guide frame, the ratchet bar is welded on the sliding support frame, the ratchet rack is characterized in that toggle rods are connected to the sliding support frames at the same position in an evenly-arranged mode, a supporting sliding block is fixedly connected to the L-shaped supporting plate, a sliding frame is connected to the supporting sliding block in a sliding mode, a pawl is connected to the sliding frame in a rotating mode, the pawl is in contact with a ratchet rack, a second torsion spring is connected between the pawl and the sliding frame, a first reset frame is connected to the ratchet rack in a sliding mode, the first reset frame is in contact with the sliding frame, a seventh spring is connected between the first reset frame and the ratchet rack, and a second reset frame is fixedly connected to the ratchet rack.
Optionally, the liquid outlet control assembly is arranged on the sliding frame and comprises a connecting frame, a wedge block and a push rod, the connecting frame is fixedly connected to the sliding frame, the wedge block is fixedly connected to the connecting frame, and the push rod is fixedly connected to the other sliding support frame.
Optionally, the system further comprises a sedimentation prevention assembly, the sedimentation prevention assembly is arranged on the reaction frame and comprises a fixed gear ring, a stirring frame and a straight gear, the fixed gear ring is fixedly arranged on the reaction frame, the stirring frame is rotatably connected to the rotating circular plate in a four-corner distribution mode, the straight gear is fixedly connected to the stirring frame, and the straight gear is meshed with the fixed gear ring.
The invention has the beneficial effects that:
through the effect that promotes the baffle, promote the baffle with the solution interval, the staff is continuous to add the mixed solution of albumen liquid and dislysate and the albumen dislysate that different concentrations arranged according to the gradient from high to low in proper order in the reaction frame, reach the mesh that can dilute the albumen dislysate in succession, the partial solution of one of them check can flow to the second carriage in the reaction frame simultaneously, guarantee to add the albumen dislysate and take out this operation conflict with solution, do not need the manual work to change the liquid, make albumen renaturation process be in a continuous state, guarantee that albumen dislysate concentration presents the linearity and subtracts progressively, be convenient for realize the albumen renaturation.
Through the cooperation of dysmorphism propelling movement frame and area card post rocker, replace artifical manual removal baffler, the mixed liquid of protein liquid and dislysate and the protein dislysate of different concentrations of continuity add to the reaction frame in, save the trouble of artifical manual mixed liquid and the protein dislysate that adds protein liquid and dislysate, realize the mixed liquid and the protein dislysate of full-automatic supply protein liquid and dislysate, provide stable liquid and dilute the protein dislysate continuously.
Rotate through stirring the frame for stirring the frame and stirring solution in to the reaction frame, with solution and albumen dislysate stirring, make solution and albumen dislysate intensive mixing, fully dilute the albumen dislysate, avoid albumen renaturation in-process to appear the albumen sediment 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 view of a first partial body structure according to the present invention.
FIG. 4 is a schematic view of a second partial body structure according to the present invention.
Fig. 5 is a perspective view of a third embodiment of the present invention.
Fig. 6 is a partially disassembled perspective structure of the present invention.
FIG. 7 is a schematic view of a first partial body configuration of the separation assembly of the present invention.
Fig. 8 is a schematic view of a partially cut-away perspective structure of the separator assembly of the present invention.
FIG. 9 is a schematic view of a second partial body configuration of the separation assembly of the present invention.
Fig. 10 is a schematic perspective view of a portion of the injection assembly of the present invention.
Figure 11 is a first partially cross-sectional isometric view of the inject assembly of the invention.
Figure 12 is a second partially cut-away perspective view of the infusion assembly of the present invention.
Figure 13 is a schematic view of a third partially cut-away perspective structure of the infusion assembly of the present invention.
FIG. 14 is a schematic diagram of a first partially assembled body of a switching assembly according to the present invention.
FIG. 15 is a perspective view of a second portion of the switching assembly of the present invention.
Fig. 16 is a perspective view of a third portion of the switching assembly of the present invention.
Fig. 17 is a perspective view of a fourth portion of the switching assembly of the present invention.
Fig. 18 is a schematic partial perspective view of a liquid control assembly according to the present invention.
Fig. 19 is a schematic perspective view of the sliding support and the push rod of the present invention.
Fig. 20 is a schematic perspective view of the anti-settling assembly of the present invention.
Fig. 21 is a schematic perspective view of a portion of the anti-settling assembly of the present invention.
Reference numbers in the figures: 1: first support frame, 21: second support, 22: reaction block, 23: motor, 24: rotating frame, 25: first support frame, 26: fixed support plate, 27: discharge pipe, 28: slide frame, 29: open-hole frame, 210: first spring, 3: agitating assembly, 31: rotating circular plate, 32: push the baffle, 4: separation assembly, 41: separation frame, 42: second support frame, 421: flow guide frame, 43: push plate, 44: second spring, 45: check valve, 46: push frame, 47: third support, 48: sliding slotted frame, 49: toggle frame, 5: injection assembly, 51: l-shaped support plate, 52: sliding support, 53: liquid storage frame, 531: input tube, 532: first aperture frame, 533: second open-hole frame, 534: third open hole frame, 54: fourth support frame, 55: first plug frame, 551: second plug rack, 552: third plug rack, 56: third spring, 57: protective folding sleeve, 58: buoyancy block, 581: barrier plate, 59: dysmorphism pushing frame, 510: fourth spring, 6: switching assembly, 61: rocker with clamp column, 6101: fifth spring, 62: fifth support frame, 63: guide frame, 631: swing plate, 632: first torsion spring, 64: sixth spring, 65: ratchet bar, 66: tap lever, 67: support slider, 68: carriage, 69: pawl, 610: second torsion spring, 611: first reset shelf, 612: seventh spring, 613: second reset shelf, 7: liquid outlet control assembly, 71: link, 72: wedge block, 73: catch bar, 8: anti-settling component, 81: fixed ring gear, 82: agitation frame, 83: a straight gear.
Detailed Description
In the description of the present invention, "a plurality" means two or more unless otherwise specified; the terms "upper", "lower", "left", "right", "inner", "outer", "front", "rear", "head", "tail", and the like, indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, are only for convenience in describing and simplifying the description, and do not indicate or imply that the device or element referred to must have a particular orientation, be constructed in a particular orientation, and be operated, and thus, 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
A continuous protein renaturation device based on enzyme engineering is shown in figures 1-10 and 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, a perforated frame 29, a first spring 210, a stirring component 3 and a separation 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 the 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 rotatably connected with the reaction frame 22, the first support frame 25 is fixedly arranged on the reaction frame 22, the fixed support plate 26 is arranged on the first support frame 25, the discharge pipe 27 is fixedly connected to the bottom of the reaction frame 22, the discharge pipe 27 is used for discharging a solution, the discharge pipe 27 is connected with a sliding frame 28 in a sliding manner, the sliding frame 28 is connected with the reaction frame 22 in a sliding manner, the discharge pipe 27 is fixedly connected with an opening frame 29, the opening frame 29 is connected with the sliding frame 28 in a sliding manner, the opening frame 29 is used for blocking the sliding frame 28, the sliding frame 28 is fixedly connected with a first spring 210, one end of the first spring 210 is fixedly connected with the discharge pipe 27, the stirring assembly 3 is arranged on the rotating frame 24, the stirring assembly 3 is used for stirring the solution in the reaction frame 22, and the separation assembly 4 is arranged on the reaction frame 22.
The stirring assembly 3 comprises a rotating circular plate 31 and a pushing partition plate 32, the rotating frame 24 is connected with the rotating circular plate 31 in a bolt connection mode, the rotating circular plate 31 is rotatably connected with the reaction frame 22, the rotating frame 24 is connected with the pushing partition plate 32 in a circumferential distribution mode, the pushing partition plate 32 is in contact with the reaction frame 22, and the pushing partition plate 32 is used for separating the solution in the reaction frame 22.
The separating assembly 4 comprises a separating frame 41, a second supporting frame 42, a flow guide frame 421, a push plate 43, a second spring 44, a one-way valve 45, a push frame 46, a third supporting frame 47, a sliding slotting frame 48 and a toggle frame 49, wherein the separating frame 41 is fixedly connected to the reaction frame 22, the separating frame 41 is provided with the second supporting frame 42, the second supporting frame 42 is fixedly connected with the reaction frame 22, the flow guide frame 421 is arranged in the second supporting frame 42, the second supporting frame 42 is connected with the push plate 43 in a sliding manner, the push plate 43 is used for extruding the solution in the second supporting 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 supporting frame 42, the push plate 43 is provided with the one-way valve 45, the push frame 46 is fixedly connected to the push plate 43, one side of the reaction frame 22 is welded with the third supporting frame 47, the third supporting frame 47 is connected with the sliding slotting frame 48 in a sliding manner, the sliding slotting bracket 48 is in limit fit with the pushing bracket 46, the rotating bracket 24 is fixedly connected with a stirring bracket 49, and the stirring bracket 49 is used for pushing the sliding slotting bracket 48 to move downwards.
When protein renaturation needs to be realized, a worker manually starts the motor 23, prepared protein solution is added into the reaction frame 22 through a liquid injection port in the fixed supporting plate 26, the partition plate 32 is pushed to separate the solution, an output shaft of the motor 23 rotates to drive the rotating frame 24 and the upper device thereof to rotate clockwise, the partition 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, the solution in one grid can be mixed with the protein dialysate, and the solution can dilute the protein dialysate. Meanwhile, the rotating frame 24 rotates to drive the stirring frame 49 to rotate, the stirring frame 49 pushes the sliding slotted frame 48 to move downwards, and the sliding slotted frame 48 pushes the pushing frame 46 and the devices thereon to move downwards, so that part of the solution in one of the cells in the reaction frame 22 flows into the second supporting 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 guide frame 421, the worker collects the solutions with different concentrations, after pushing the partition plate 32 to rotate for a circle, the worker adds the protein dialysate with lower concentration into the reaction frame 22, the operation is repeated, the worker continuously adds the protein dialysate with different concentrations into the reaction frame 22 from high to low according to the gradient, the protein dialysate can be continuously diluted, the operation of adding the protein dialysate and extracting the solution is not conflicted, 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 to 14, the apparatus further comprises an injection component 5, the injection component 5 is fixedly installed on the top surface of the fixed support plate 26, the injection component 5 is used for adding protein dialysate into the reaction frame 22, the injection component 5 comprises an L-shaped support plate 51, a sliding support frame 52, a liquid storage frame 53, an input tube 531, a first open-hole frame 532, a second open-hole frame 533, a third open-hole 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 baffle frame 58, a baffle 581, a special-shaped pushing frame 59 and a fourth spring 510, the two L-shaped support plates 51 are fixedly installed on the top surface of the fixed support plate 26, the sliding support frame 52 is slidably connected to the L-shaped support plate 51, the three liquid storage frames 53 are fixedly connected to the two sliding support frames 52, the input tube 531 is installed on the liquid storage frame 53, wherein a first open-hole frame 532 is arranged on one liquid storage frame 53, a second open-hole frame 533 is arranged on the other liquid storage frame 53 far away from the first open-hole frame 532, a third open-hole frame 534 is arranged on the other liquid storage frame 53, a fourth support frame 54 is arranged in the liquid storage frame 53, wherein a first blocking frame 55 is slidably connected with one fourth support frame 54, the first blocking frame 55 is in contact with the first open-hole frame 532, the first blocking frame 55 is used for blocking the liquid outlet on the first open-hole frame 532, a second blocking frame 551 is slidably connected with the other fourth support frame 54, the second blocking frame 551 is in contact with the second open-hole frame 533, the second blocking frame 551 is used for blocking the liquid outlet on the second open-hole frame 533, a third blocking frame 552 is slidably connected with the other fourth support frame 54, the third blocking frame 552 is in contact with the third open-hole frame 534, and the third blocking frame 552 is used for blocking the liquid outlet on the third open-hole frame 534, a third spring 56 is connected between the first blocking frame 55 and one fourth supporting frame 54, a third spring 56 is connected between the second blocking frame 551 and one of the fourth supporting frames 54, a third spring 56 is connected between the third blocking frame 552 and the other fourth supporting frame 54, a protective folding sleeve 57 is fixedly connected to the fourth supporting frame 54, one protective folding sleeve 57 is fixedly connected to the first open-hole frame 532, the other protective folding sleeve 57 is fixedly connected to the second open-hole frame 533, the other protective folding sleeve 57 is fixedly connected to the third open-hole frame 534, a buoyancy blocking frame 58 is slidably connected to the liquid storage frame 53, a blocking plate 581 is commonly arranged on the three liquid storage frame 53, the first open-hole frame 532, the second open-hole frame 533 and the third open-hole frame 534 are fixedly connected to the blocking plate 581, the blocking plate 581 is in contact with the fixed supporting plate 26, the blocking plate 581 is in contact with the special-shaped supporting plate 26, a special-shaped pushing frame 59 is slidably connected to the fixed supporting plate 26, and used for pushing the second blocking frame 551, The third blocking frame 552 or the first blocking frame 55 moves upward, and a fourth spring 510 is connected between the fixed support plate 26 and the deformed pushing frame 59.
The staff adds the mixed solution of albumen liquid and dislysate and the albumen dislysate that different concentrations arranged according to the gradient in proper order 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 albumen liquid and dislysate produces buoyancy and promotes buoyancy and keep off frame 58 upward movement, and buoyancy keeps off frame 58 and blocks the inlet on the stock solution frame 53 gradually for the inlet fluid speed reduces according to the liquid level risees gradually. Then, the worker manually pushes the blocking plate 581 toward the direction close to the deformed pushing frame 59 so that the second open frame 533 corresponds to the pouring port of 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 pushes the second blocking frame 551 to move upwards, and the second blocking frame 551 does not block the liquid outlet hole in the second open-hole frame 533 any more, so that the mixed liquid of the protein liquid and the dialysate 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 for resetting, and the compressed third spring 56 resets to drive the second blocking frame 551 to move downwards for resetting, so that the second blocking frame 551 blocks the liquid outlet hole on the second open hole frame 533. The above operation was repeated, and the 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 toward the special-shaped pushing frame 59 again, 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 no longer blocks the liquid outlet hole of the third open-hole 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 in the third open hole frame 534.
After the toggle frame 49 rotates one more turn, the worker pushes the blocking plate 581 again toward the special-shaped pushing frame 59, so that the first opening frame 532 corresponds to the liquid injection port of 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 no longer blocks the liquid outlet hole of the first hole-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 hole opening frame 532. By repeating the above operation, the mixed liquid of the protein liquid and the dialysate and the protein dialysate with different concentrations are continuously added into the reaction frame 22, so that the trouble of manually adding the mixed liquid of the protein liquid and the dialysate and the protein dialysate is eliminated, and the stable liquid is provided for continuously diluting the protein dialysate.
Example 3
Based on the embodiment 2, as shown in fig. 14 to 17, the apparatus further includes a switching assembly 6, the first supporting frame 25 is provided with the switching assembly 6, the switching assembly 6 is used for switching the concentration of the added protein dialysate, the switching assembly 6 includes a rocker 61 with a clamping column, a fifth spring 6101, a fifth supporting frame 62, a guiding frame 63, a swinging plate 631, a first torsion spring 632, a sixth spring 64, a ratchet rack 65, a poking rod 66, a supporting slider 67, a sliding frame 68, a pawl 69, a second torsion spring 610, a first resetting frame 611, a seventh spring 612, and a second resetting frame 613, the rotating frame 24 is fixedly connected with the rocker 61 with the clamping column, the fifth spring 6101 is connected between the L-shaped supporting plate 51 and the sliding supporting frame 52, the first supporting frame 25 is fixedly connected with the fifth supporting frame 62, the guiding frame 63 is slidably connected to the fifth supporting frame 62, the sixth spring 64 is connected to the fifth supporting frame 62, one end of a sixth spring 64 far away from the fifth support frame 62 is connected with the guide frame 63, the guide frame 63 near the fifth support frame 62 is rotatably connected with a swinging plate 631, a first torsion spring 632 is connected between the swinging plate 631 and the guide frame 63, the ratchet rack 65 is welded on a sliding support frame 52, the sliding support frame 52 is connected with toggle rods 66 in a uniform arrangement manner, an L-shaped support plate 51 is fixedly connected with a support slider 67, the support slider 67 is slidably connected with a sliding frame 68 in a sliding manner, the sliding frame 68 is rotatably connected with a pawl 69, the pawl 69 is contacted with the ratchet rack 65, the pawl 69 is used for clamping the ratchet rack 65, a second torsion spring 610 is connected between the pawl 69 and the sliding frame 68, the ratchet rack 65 is slidably connected with a first reset frame 611, the first reset frame 611 is contacted with the sliding frame 68, the first reset frame 611 is used for pushing the sliding frame 68 and the upper device thereof to move towards the direction close to the sliding support frame 52, a seventh spring 612 is connected between the first reset rack 611 and the ratchet rack 65, and a second reset rack 613 is fixedly connected to the ratchet rack 65 far away from the first reset rack 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 device thereon to move upwards, the swing plate 631 pushes the poke rod 66 and the device thereon to move towards the direction close to the special-shaped pushing frame 59, the barrier 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, the pawl 69 clamps the ratchet rack 65 under the action of the second torsion spring 610, and the fifth spring 6101 is prevented from resetting to drive the sliding support frame 52 and the device thereon to move towards the direction away from the special-shaped pushing frame 59.
When the rotating frame 24 is separated from the rocker 61 with the clamping column, the compressed sixth spring 64 resets to drive the guide frame 63 and the device thereon to move downwards, the poke 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 and the sliding frame 68 contact each other, the second reset frame 613 pushes the sliding frame 68 and the upper device to move away from the sliding support frame 52, so that the pawl 69 no longer blocks the ratchet bar 65, the extended fifth spring 6101 resets to drive the sliding support frame 52 and the upper device to move away from the special-shaped pushing frame 59 for resetting, and then the first reset frame 611 pushes the sliding frame 68 and the upper device to move towards the sliding support frame 52, and the pawl 69 blocks the ratchet bar 65 again.
Example 4
On the basis of the embodiment 3, as shown in fig. 18 to 19, the liquid discharge control device 7 is further included, the liquid discharge control device 7 is disposed on the sliding frame 28, the liquid discharge control device 7 is used for controlling the discharge pipe 27 to discharge the solution, the liquid discharge control device 7 includes a connecting frame 71, a wedge-shaped block 72 and a push rod 73, the connecting frame 71 is fixedly connected to the sliding frame 28, the connecting frame 71 far away from the sliding frame 28 is fixedly connected to the wedge-shaped block 72, and the other sliding support frame 52 is fixedly connected to the push rod 73.
When the sliding support frame 52 and the upper device thereof move towards the direction close to the special-shaped pushing frame 59, the pushing rod 73 contacts the wedge block 72, the pushing rod 73 pushes the wedge block 72 and the connecting frame 71 to move downwards, the connecting frame 71 drives the sliding frame 28 to move downwards, so that the hole opening frame 29 does not block the sliding frame 28 any more, and the solution in the reaction frame 22 is discharged from the discharge pipe 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 completely discharged from the discharge pipe 27. When the sliding support frame 52 and the upper device thereof move away from the special-shaped pushing frame 59, the pushing rod 73 is separated from the wedge block 72, the compressed first spring 210 is reset to drive the sliding frame 28 to move upwards for resetting, and the hole frame 29 blocks the sliding frame 28.
Example 5
On the basis of embodiment 4, as shown in fig. 20 to 21, the system further comprises an anti-sedimentation assembly 8, the anti-sedimentation assembly 8 is disposed on the reaction frame 22, the anti-sedimentation assembly 8 is used for preventing protein sedimentation during protein renaturation, the anti-sedimentation assembly 8 comprises a fixed gear ring 81, a stirring frame 82 and a spur gear 83, the fixed gear ring 81 is fixedly mounted on the reaction frame 22, the stirring frame 82 is rotatably connected to the rotating circular plate 31 in a four-corner distribution manner, the stirring frame 82 is used for stirring a solution in the reaction frame 22, the spur gear 83 is fixedly connected to the stirring frame 82, and the spur gear 83 is meshed with the fixed gear ring 81.
The rotating frame 24 can drive the rotating circular plate 31 and the devices thereon to rotate, and under the action of the fixed gear ring 81, the straight gear 83 and the stirring frame 82 rotate, so that the stirring frame 82 can stir the solution in the reaction frame 22, the solution and the protein dialysate can be fully mixed, the protein dialysate can be fully diluted, and protein precipitation in the protein renaturation process can be avoided.
The embodiments of the present 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 embodiment was 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 (8)
1. The utility model provides a continuous type protein renaturation device based on enzyme engineering which characterized in that, including first support frame, second support frame, reaction frame, motor, rotating turret, first support frame, fixed support board, discharge pipe, slide frame, trompil frame, first spring, stirring subassembly and separator assembly:
the first support frame is connected with the second support frame in a fastener connection mode;
the reaction frame is fixedly arranged on the first support frame and used for providing a reaction site for protein renaturation;
the motor is fixedly arranged at the top of the second support frame and used for providing driving power for equipment;
one end of an output shaft of the motor is welded with the rotating frame, and the rotating frame is rotatably connected with the reaction frame;
the reaction frame is fixedly provided with a first support frame;
the first support frame is provided with a fixed support plate, and the fixed support plate is used for adding protein solution or protein dialysate;
the discharge pipe is fixedly connected to the bottom of the reaction frame and is used for discharging the protein renaturation solution;
the discharge pipe is connected with the sliding frame in a sliding way, and the sliding frame is connected with the reaction frame in a sliding way;
the discharge pipe is fixedly connected with the tapping frame, the tapping frame is connected with the sliding frame in a sliding mode, and the tapping frame is used for plugging 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 component 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 the protein solution in the reaction frame at intervals.
2. The continuous protein renaturation device according to the enzyme engineering as claimed in claim 1, wherein the stirring assembly comprises a rotating circular plate and a pushing partition plate, the rotating circular plate is connected to the rotating frame by means of bolts, the rotating circular plate is rotatably connected to the reaction frame, the pushing partition plates are connected to the rotating frame in a circumferentially distributed manner, and the pushing partition plates are in contact with the reaction frame.
3. The continuous protein renaturation device based on enzyme engineering according to claim 2 characterized in that four pushing clapboards are arranged on the rotating frame, the pushing clapboards are closely attached to the inner wall of the reaction frame, the pushing clapboards are used for separating protein liquids with different concentrations in the reaction frame, the pushing clapboards can push the solution in the reaction frame to flow, and it is fully ensured that the solution is not conflicted with the protein dialysate.
4. The continuous protein renaturation device based on enzyme engineering as claimed in claim 2, the separation assembly comprises a separation frame, a second supporting frame, a flow guide frame, a push plate, a second spring, a one-way valve, a push frame, a third supporting frame, a sliding slotting frame and a poking frame, wherein the separation frame is fixedly connected to the reaction frame, the second supporting frame is arranged on the separation frame, the second supporting frame is fixedly connected with the reaction frame, the flow guide frame is arranged in the second supporting frame, the push plate is slidably connected to the second supporting frame, the second spring is fixedly connected to the push plate, one end of the second spring is fixedly connected to the second supporting frame, the one-way valve is arranged on the push plate, the push 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 slidably connected to the third supporting frame, the sliding slotting frame is in limited fit with the push frame, and the poking frame is fixedly connected to the rotating frame.
5. The continuous protein renaturation device according to the enzyme engineering based on the claim 4, characterized in that it further comprises an injection component, the injection component is fixedly installed 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 tube, a first open hole frame, a second open hole frame, a third open hole frame, a fourth support frame, a first blocking frame, a second blocking frame, a third spring, a protective folding sleeve, a buoyancy blocking frame, a blocking plate, a special-shaped pushing frame and a fourth spring, two L-shaped support plates are fixedly installed on the top surface of the fixed support plate, the sliding support frames are slidably connected with each other, the three liquid storage frames are fixedly connected with each other on the two sliding support frames, the input tube is installed on the liquid storage frame, the first open hole frame is installed on one liquid storage frame, the second open hole frame is installed on the other liquid storage frame, a third open pore frame is arranged on the other liquid storage frame, a fourth support frame is arranged in the liquid storage frame, wherein a first blocking frame is connected on the fourth support frame in a sliding way, the first blocking frame is contacted with the first open pore frame, a second blocking frame is connected on the other fourth support frame in a sliding way, the second blocking frame is contacted with the second open pore frame, a third blocking frame is connected on the other fourth support frame in a sliding way, the third blocking frame is contacted with the third open pore frame, a third spring is connected between the first blocking frame and one fourth support frame, a third spring is connected between the second blocking frame and one fourth support frame, a third spring is connected between the third blocking frame and the other fourth support frame, a protective folding sleeve is fixedly connected on the fourth support frame, one protective folding sleeve is fixedly connected with the first open pore frame, the other protective folding sleeve is fixedly connected with the second open pore frame, the other protective folding sleeve is fixedly connected with the third open pore frame, the liquid storage frame is connected with a buoyancy blocking frame in a sliding mode, the three liquid storage frames are provided with a blocking plate together, the first opening frame, the second opening frame and the third opening frame are fixedly connected with the blocking plate, the blocking plate is in contact with the fixed supporting plate, the special-shaped pushing frame is connected to the fixed supporting plate in a sliding mode, and a fourth spring is connected between the fixed supporting plate and the special-shaped pushing frame.
6. The continuous protein renaturation device according to the enzyme engineering as claimed in claim 5, further comprising a switching assembly, wherein said switching assembly is disposed on said first support frame, said switching assembly comprises a rocker with a locking pin, a fifth spring, a fifth support frame, a guide frame, a swing plate, a first torsion spring, a sixth spring, a ratchet bar, a toggle bar, a support slider, a sliding frame, a pawl, a second torsion spring, a first reset frame, a seventh spring and a second reset frame, said rotating frame is fixedly connected with said rocker with a locking pin, a fifth spring is connected between said L-shaped support frame and said sliding support frame, said fifth support frame is fixedly connected with said fifth support frame, said fifth support frame is slidably connected with said guide frame, said fifth support frame is connected with said sixth spring, one end of said sixth spring is connected with said guide frame, said guide frame is rotatably connected with said swing plate, the ratchet rack is welded on a sliding support frame, a poking rod is connected on the sliding support frame at the same position in an evenly-arranged mode, a supporting sliding block is fixedly connected on an L-shaped supporting plate, a sliding frame is connected on the supporting sliding block in a sliding mode, a pawl is connected on the sliding frame in a rotating mode, the pawl is in contact with the ratchet rack, a second torque spring is connected between the pawl and the sliding frame, a first reset frame is connected on the ratchet rack in a sliding mode, the first reset frame is in contact with the sliding frame, a seventh spring is connected between the first reset frame and the ratchet rack, and a second reset frame is fixedly connected on the ratchet rack.
7. The continuous protein renaturation device based on enzyme engineering as claimed in claim 6, characterized in that it further comprises a liquid outlet control assembly, said liquid outlet control assembly is mounted on the sliding frame, said liquid outlet control assembly comprises a connection frame, a wedge block and a push rod, said connection frame is fixedly connected to the sliding frame, said wedge block is fixedly connected to the connection frame, and said push rod is fixedly connected to another sliding support frame.
8. The continuous protein renaturation device based on enzyme engineering according to claim 7, characterized in that it further comprises a precipitation prevention assembly, wherein the precipitation prevention assembly is arranged on the reaction frame, the precipitation prevention assembly comprises a fixed gear ring, a stirring frame and a spur gear, the fixed gear ring is fixedly arranged on the reaction frame, the stirring frame is rotatably connected to the rotating circular plate in a four-corner distribution manner, the spur gear is fixedly connected to the stirring frame, and the spur gear is meshed with the fixed gear ring.
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