CN112979745A - Method for purifying antibacterial peptide by solid phase extraction - Google Patents
Method for purifying antibacterial peptide by solid phase extraction Download PDFInfo
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- CN112979745A CN112979745A CN202110473220.6A CN202110473220A CN112979745A CN 112979745 A CN112979745 A CN 112979745A CN 202110473220 A CN202110473220 A CN 202110473220A CN 112979745 A CN112979745 A CN 112979745A
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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
- C07K1/14—Extraction; Separation; Purification
- C07K1/145—Extraction; Separation; Purification by extraction or solubilisation
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
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Abstract
The invention discloses a method for purifying antibacterial peptide by solid phase extraction, which comprises the following steps: step 1, preparing materials; a raw material containing an antimicrobial peptide is prepared. Step 2, degreasing; adding lipase into the raw material containing the antibacterial peptide for degreasing. Step 3, extracting; and (3) extracting the degreased raw materials in an extraction device, standing for layering, and taking down the lower-layer solution, wherein the temperature in the extraction device is kept at 37 ℃, the mixing time is 2-2.5 hours, and the standing time is 40-60 minutes. The extraction device comprises: extraction jar, actuating mechanism and mix the assembly. The method for purifying the antibacterial peptide by solid-phase extraction is simple in operation, and the adopted extraction device can quickly mix the raw materials, so that the internal raw materials can quickly react under the mixing of the mixing assembly, and then the reaction that the solution in the raw materials can be sufficient is ensured. Quickening the reaction efficiency, shortening the time and improving the quality of the purified antibacterial peptide.
Description
Technical Field
The invention relates to the technical field of antibacterial peptide purification, in particular to a method for purifying antibacterial peptide by solid-phase extraction.
Background
The antibacterial peptide generally consists of 12-100 amino acid residues, mostly contains hydrophobic amino acids, and mostly is amphiphilic cationic peptide. The natural antibacterial peptide has wide antibacterial spectrum, good thermal stability and water solubility, almost no toxic action on normal cells of higher animals, and is not easy to generate drug resistance. Research on antimicrobial peptides of different species has been a hot spot, and the antimicrobial peptides have the potential to be used as a new generation of antimicrobial drugs to replace traditional antibiotics.
However, the extraction device adopted at present is simple in structure, long in mixing time and incapable of controlling the mixing efficiency of the whole mixing structure. Resulting in a decrease in extraction efficiency and difficulty in extracting antibacterial peptides of good quality.
Disclosure of Invention
In order to solve the defects of the prior art, the invention provides a method for purifying antibacterial peptide by solid-phase extraction.
The technical scheme of the invention is realized as follows:
a method for purifying antibacterial peptide by solid phase extraction comprises the following steps:
step 1, preparing materials; preparing a raw material containing antibacterial peptide;
the extraction device comprises:
the extraction tank is used for placing raw materials containing the antibacterial peptide, the upper end of the extraction tank is provided with a liquid inlet pipe, and the lower end of the extraction tank is provided with a liquid outlet pipe;
the driving mechanism is arranged at the upper end of the extraction tank, a motor, a driving rod and a gear are arranged in the driving mechanism, the motor is arranged on the extraction tank through a fixing frame, a first rotating shaft is arranged at the lower end of the motor, a rotating piece is arranged on the first rotating shaft and is hinged with one end of the driving rod, a sawtooth part is arranged on the driving rod, the gear is arranged on the extraction tank, a pushing piece is arranged on the extraction tank and is arranged on the extraction tank through a first hinged rod, a pushing rod is arranged on the pushing piece, one end of the pushing rod is hinged on the pushing piece, the other end of the pushing rod is provided with a pushing piece, one end of the pushing piece is hinged with the pushing rod, the middle of the pushing piece is arranged on the extraction tank through a second hinged rod, the other end of the pushing piece is contacted with the driving rod, and a rotating part is arranged at the lower end, the periphery of the rotating part is provided with triangular blades;
mix the assembly, mix the inside at the extraction jar is installed to the assembly, first rabbling mechanism and second rabbling mechanism have in mixing the assembly, first rabbling mechanism and the same but rotational symmetry setting of second rabbling mechanism structure, the rotary rod has in the first rabbling mechanism, rotary rod and gear connection, first rotatory leaf has on the rotary rod, the lower extreme of rotary rod has the rotary disk, the lower extreme of rotary disk has the second rotatory leaf, the pull rod has on the rotary disk, the fixed block has on the inner wall of extraction jar, the slurcam has on the fixed block, the slurcam articulates on the fixed block, the one end of pull rod articulates on the rotary disk, and the other end articulates on the slurcam.
In the invention, the driving rod is also provided with a limiting sliding groove, and one end of the pushing piece is arranged in the limiting sliding groove.
In the invention, the pushing piece is provided with a sliding column, the sliding column is sleeved with a limiting piece, two ends of the limiting piece are provided with limiting parts, the limiting parts are provided with limiting arc grooves, and the extraction tank is provided with a first limiting rod and a second limiting rod.
In the invention, the first limiting rod and the second limiting rod have equal distance to the first hinge rod.
In the invention, the sliding column is also sleeved with a first spring, one end of the first spring is connected with the lower end of the limiting piece, and the other end of the first spring is connected with the pushing piece.
In the invention, the extraction tank is provided with an installation column, the driving rod is provided with an installation block, and the installation column and the installation block are connected through a second spring.
In the present invention, the first rotary vane has a corrugated cross-section.
In the invention, the pull rod is provided with a groove.
In the invention, the first stirring mechanism and the second stirring mechanism are driven by the second rotating blade.
In the invention, the pushing plate is arc-shaped and is in tangential fit with the inner wall of the extraction tank.
The method for purifying the antibacterial peptide by solid phase extraction has the following beneficial effects: the method for purifying the antibacterial peptide by solid-phase extraction is simple in operation, and the adopted extraction device can quickly mix the raw materials, so that the internal raw materials can quickly react under the mixing of the mixing assembly, and then the reaction that the solution in the raw materials can be sufficient is ensured. Quickening the reaction efficiency, shortening the time and improving the quality of the purified antibacterial peptide.
Simultaneously through pushing away the drive that the piece can control mixing assembly, guarantee that only the lobe can rotate at the anterior segment part in the stage of stewing, then when arriving the back end part in the stage of stewing, close the motor again, guarantee that the solution lower extreme after the extraction can not receive the stirring influence, and the lobe can carry out slight stirring with the solution of upper end, guarantees that the solution of upper end can not appear condensing phenomenon.
Drawings
FIG. 1 is a schematic view of an extraction apparatus according to the present invention;
FIG. 2 is a schematic view of the internal structure of FIG. 1;
FIG. 3 is a schematic view of the drive mechanism of FIG. 2;
FIG. 4 is another schematic view of the structure of FIG. 3;
FIG. 5 is a schematic view of the pushing member shown in FIG. 4;
FIG. 6 is a schematic diagram of the hybrid assembly of FIG. 1;
fig. 7 is a schematic structural diagram of the first stirring mechanism in fig. 6.
In the figure: the device comprises an extraction device 1, an extraction tank 2, a liquid inlet pipe 3, a liquid discharge pipe 4, a driving mechanism 5, a motor 6, a driving rod 7, a gear 8, a first rotating shaft 9, a rotating part 10, a sawtooth part 11, a limiting sliding chute 12, a pushing part 13, a first hinged rod 14, a pushing rod 15, a pushing part 16, a second hinged rod 17, a rotating part 18, a triangular blade 19, a sliding column 20, an installation column 201, an installation block 202, a limiting part 21, a limiting part 22, a first limiting rod 23, a second limiting rod 24, a first spring 25, a second spring 26, a limiting arc groove 27, a mixing assembly 28, a first stirring mechanism 281, a second stirring mechanism 282, a rotating rod 29, a first rotating blade 30, a second rotating blade 31, a rotating disk 32, a pull rod 33, a groove 34, a fixed block 35 and a pushing plate 36.
Detailed Description
The technical solution in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention.
As shown in fig. 1 to 7, the method for purifying antimicrobial peptide by solid phase extraction of the present invention comprises the following steps:
step 1, preparing materials; a raw material containing an antimicrobial peptide is prepared.
The extraction apparatus 1 comprises:
and the extraction tank 2 is used for placing the raw materials containing the antibacterial peptides. The upper end of the extraction tank 2 is provided with a liquid inlet pipe 3, and the lower end is provided with a liquid discharge pipe 4. The liquid inlet pipe 3 is used for feeding raw materials, and the liquid outlet pipe 4 is used for discharging the extracted solution.
And the driving mechanism 5 is arranged at the upper end of the extraction tank 2. The driving mechanism 5 has therein a motor 6, a driving lever 7 and a gear 8. The motor 6 is installed on the extraction tank 2 through a fixing frame (not shown in the figure), the lower end of the motor 6 is provided with a first rotating shaft 9, a rotating piece 10 is arranged on the first rotating shaft 9, the rotating piece 10 is hinged with one end of a driving rod 7, a sawtooth part 11 is arranged on the driving rod 7, the gear 8 is installed on the extraction tank 2, and the sawtooth part 11 can be meshed with the gear 8 after being pushed.
The driving rod 7 is also provided with a limit sliding groove 12, one end of a pushing piece 16 is arranged in the limit sliding groove 12, and the limit sliding groove 12 is used for limiting the position of the pushing rod 15.
The extraction tank 2 is provided with a pushing piece 13, the pushing piece 13 is installed on the extraction tank 2 through a first hinged rod 14, and the pushing piece 13 is provided with a pushing rod 15. One end of a push rod 15 is hinged on the push piece 13, the other end of the push rod is provided with a push piece 16, one end of the push piece 16 is hinged with the push rod 15, the middle of the push piece 16 is installed on the extraction tank 2 through a second hinge rod 17, and the other end of the push piece 16 is contacted with the driving rod 7.
The first rotating shaft 9 has a rotating portion 18 at a lower end thereof, and a triangular blade 19 is provided around the rotating portion 18. The triangular blade 19 is located inside the extraction tank 2, and at the upper end of the extraction tank 2, the raw materials can be mixed and extracted, the solution at the upper end is slightly mixed and stirred by the front section part of the standing stage, the condensation phenomenon of the solution at the upper end after layering is avoided, and the triangular blade 19 can avoid the phenomenon.
The pushing piece 13 is provided with a sliding column 20, the sliding column 20 is sleeved with a limiting piece 21, two ends of the limiting piece 21 are provided with limiting parts 22, the limiting parts 22 are provided with limiting arc grooves 27, and the extraction tank 2 is provided with a first limiting rod 23 and a second limiting rod 24. By pulling the limiting member 21, the first spring 25 can be stretched, and then the pushing member 13 is pushed, so that the pushing member 13 pushes the pushing rod 15, and then the pushing member 16 is pushed by the pushing rod 15, and then the driving rod 7 is pressed downwards by the other end of the pushing member 16, so that the sawtooth part 11 is connected with the gear 8 in a meshing manner.
The first and second limit levers 23 and 24 are equidistant from the first hinge lever 14. When the pushing piece 13 is rotated, the limiting arc groove 27 on the limiting part 22 can be arranged on the second limiting rod 24, so that the pushing piece 13 is limited.
Meanwhile, the sliding column 20 is sleeved with a first spring 25, one end of the first spring 25 is connected with the lower end of the limiting part 21, and the other end of the first spring 25 is connected with the pushing part 13.
The extraction tank 2 is provided with a mounting column 201, the driving rod 7 is provided with a mounting block 202, and the mounting column 201 and the mounting block 202 are connected through a second spring 26.
When the pusher 13 is rotated onto the first limit rod 23, the drive rod 7 can be pulled back by means of the second spring 26, so that the toothed segment 11 is disengaged from the gear wheel 8, so that the mixing assembly 28 is no longer mixed.
The mixing assembly 28 is installed inside the extraction tank 2, the mixing assembly 28 has a first stirring mechanism 281 and a second stirring mechanism 282 therein, and the first stirring mechanism 281 and the second stirring mechanism 282 are identical in structure but are arranged rotationally symmetrically. The first stirring mechanism 281 is provided with a rotating rod 29, the rotating rod 29 is connected with the gear 8, and the rotating rod 29 is provided with a first rotating blade 30. The first rotary vane 30 has a corrugated shape in cross section. When the cross section of the first rotating blade 30 is set to be corrugated, when the first rotating blade 30 rotates, the solution at the upper end can be brought to the lower end through the corrugated structure, and then the solution at the lower end can be brought to the upper end during reverse rotation, so that the solution in the whole extraction tank 2 is stirred, the mixing efficiency of the solution is accelerated, the time is shortened, and the mixing quality is improved.
The lower end of the rotating lever 29 has a rotating disk 32, and the lower end of the rotating disk 32 has a second rotating blade 31. The second rotating blade 31 can mix the solution at the lower end of the rotating disk 32, that is, the solution at the middle and lower ends of the extraction tank 2 can be mixed, so that the solution at the upper and lower ends of the extraction tank 2 can be mixed.
The rotary disc 32 has a pull rod 33, and the pull rod 33 has a groove 34, and the groove 34 is used for avoiding the rotary rod 29. The inner wall of the extraction tank 2 is provided with a fixed block 35, the fixed block 35 is provided with a push plate 36, the push plate 36 is hinged on the fixed block 35, one end of a pull rod 33 is hinged on the rotating disc 32, and the other end is hinged on the push plate 36. When the rotating disc 32 rotates, the pull rod 33 is enabled to be pulled, and then the push plate 36 is pulled by the pull rod 33, so that the solution enters into the gap between the push plate 36 and the inner wall of the extraction tank 2. When the pull rod 33 is pushed, the pull rod 33 pushes the push plate 36 again, so that the push plate 36 pushes out the solution in the gap between the inner walls of the extraction tank 2, and pushes the solution against the second rotary blade 31.
And when the pull rod 33 is pulled after the second rotating blade 31 rotates counterclockwise, the second rotating blade 31 can push the solution in the extraction tank 2 into the gap between the push plate 36 and the inner wall of the extraction tank 2, i.e. the push plate 36 is pulled to rotate, and the gap is opened. When the second rotating blade 31 rotates clockwise, the pull rod 33 can be pushed, so as to close the gap between the pushing plate 36 and the inner wall of the extraction tank 2, and the second rotating blade 31 at this time can take out the solution pushed out by the pushing plate 36 and drive the solution in the direction of the second stirring mechanism 282.
The first stirring mechanism 281 and the second stirring mechanism 282 are driven by the second rotating blade 31, and the second rotating blade 31 on the second stirring mechanism 282 can be driven by the second rotating blade 31 on the first stirring mechanism 281, so that the second stirring mechanism 282 can also drive the rotating disk 32 in the second stirring mechanism 282 to rotate, and the solution is stirred by the pushing plate 36.
And, the pushing plate 36 is arc-shaped and is in tangential contact with the inner wall of the extraction tank 2. This enables the solution in the gap between the push plate 36 and the inner wall of the extraction tank 2 to be pressed out after the push plate 36 is pushed.
Because the driving rod 7 is hinged to the rotating member 10, when the rotating member 10 is driven by the first rotating shaft 9 to rotate, the rotating member 10 pushes the driving rod 7, but the rotating member 10 does circular motion, so the driving rod 7 is pushed back and forth by the rotating member 10, the gear 8 also rotates forward and backward, the rotating rod 29 at the lower end is driven to rotate forward and backward, and the raw materials in the extraction tank 2 are mixed uniformly.
When the extraction device 1 works, the raw materials are firstly put into the extraction tank 2, and then the motor 6 is driven, so that the motor 6 drives the triangular blade 19 to rotate, and the solution at the upper end is stirred. When it is desired to mix the raw materials in the entire extraction tank 2 with agitation. The limiting member 21 is pulled, and then the pushing member 13 is pushed, so that the limiting portion 22 is in contact with the second limiting rod 24, so that the pushing member 13 pushes the pushing rod 15, and then the pushing member 16 is pushed by the pushing rod 15, so that the other end of the pushing member 16 presses the driving rod 7 downwards, and the sawtooth portion 11 is in meshed connection with the gear 8. The gear 8 rotates at this time, and then the rotating rod 29 is driven to rotate by the gear 8, so that the mixing assembly 28 in the extraction tank 2 is driven to mix.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principles of the present invention are intended to be included within the scope of the present invention.
Claims (10)
1. A method for purifying antibacterial peptide by solid phase extraction is characterized by comprising the following steps:
step 1, preparing materials; preparing a raw material containing antibacterial peptide;
step 2, degreasing; adding lipase into the raw material containing the antibacterial peptide for degreasing;
step 3, extracting; extracting the degreased raw materials in an extraction device, standing for layering, and taking the lower-layer solution, wherein the temperature in the extraction device is kept at 37 ℃, the mixing time is 2-2.5 hours, and the standing time is 40-60 minutes;
the extraction device comprises:
the extraction tank is used for placing raw materials containing the antibacterial peptide, the upper end of the extraction tank is provided with a liquid inlet pipe, and the lower end of the extraction tank is provided with a liquid outlet pipe;
the driving mechanism is arranged at the upper end of the extraction tank, a motor, a driving rod and a gear are arranged in the driving mechanism, the motor is arranged on the extraction tank through a fixing frame, a first rotating shaft is arranged at the lower end of the motor, a rotating piece is arranged on the first rotating shaft and is hinged with one end of the driving rod, a sawtooth part is arranged on the driving rod, the gear is arranged on the extraction tank, a pushing piece is arranged on the extraction tank and is arranged on the extraction tank through a first hinged rod, a pushing rod is arranged on the pushing piece, one end of the pushing rod is hinged on the pushing piece, the other end of the pushing rod is provided with a pushing piece, one end of the pushing piece is hinged with the pushing rod, the middle of the pushing piece is arranged on the extraction tank through a second hinged rod, the other end of the pushing piece is contacted with the driving rod, and a rotating part is arranged at the lower end, the periphery of the rotating part is provided with triangular blades;
mix the assembly, mix the inside at the extraction jar is installed to the assembly, first rabbling mechanism and second rabbling mechanism have in mixing the assembly, first rabbling mechanism and the same but rotational symmetry setting of second rabbling mechanism structure, the rotary rod has in the first rabbling mechanism, rotary rod and gear connection, first rotatory leaf has on the rotary rod, the lower extreme of rotary rod has the rotary disk, the lower extreme of rotary disk has the second rotatory leaf, the pull rod has on the rotary disk, the fixed block has on the inner wall of extraction jar, the slurcam has on the fixed block, the slurcam articulates on the fixed block, the one end of pull rod articulates on the rotary disk, and the other end articulates on the slurcam.
2. The method of claim 1, wherein the driving rod further comprises a position-limiting sliding groove, and one end of the pushing element is disposed in the position-limiting sliding groove.
3. The method of claim 1, wherein the pushing element has a sliding column, the sliding column has a limiting element, the two ends of the limiting element have a limiting portion, the limiting portion has a limiting arc groove, and the extraction tank has a first limiting rod and a second limiting rod.
4. The method of claim 3, wherein the first and second stop levers are equidistant from the first hinge lever.
5. The method for purifying antimicrobial peptide by solid phase extraction according to claim 3, wherein the slide column is further sleeved with a first spring, one end of the first spring is connected with the lower end of the limiting member, and the other end of the first spring is connected with the pushing member.
6. The method for purifying antimicrobial peptide by solid phase extraction as claimed in claim 1, wherein the extraction tank is provided with a mounting column, the driving rod is provided with a mounting block, and the mounting column and the mounting block are connected by a second spring.
7. The method for the solid-phase extraction purification of antimicrobial peptide according to claim 1, wherein the cross section of the first rotating blade is corrugated.
8. The method for purifying antibacterial peptide by solid phase extraction as claimed in claim 1, wherein the pull rod is provided with a groove.
9. The method for the solid-phase extraction and purification of the antibacterial peptide according to claim 1, wherein the first stirring mechanism and the second stirring mechanism are driven by a second rotating blade.
10. The method of claim 1, wherein the pushing plate is curved and is tangentially attached to the inner wall of the extraction tank.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110473220.6A CN112979745A (en) | 2021-04-29 | 2021-04-29 | Method for purifying antibacterial peptide by solid phase extraction |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110473220.6A CN112979745A (en) | 2021-04-29 | 2021-04-29 | Method for purifying antibacterial peptide by solid phase extraction |
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| CN112979745A true CN112979745A (en) | 2021-06-18 |
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| CN202110473220.6A Withdrawn CN112979745A (en) | 2021-04-29 | 2021-04-29 | Method for purifying antibacterial peptide by solid phase extraction |
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| CN113813643A (en) * | 2021-10-14 | 2021-12-21 | 江西劲农作物保护有限公司 | Method and extraction device for extracting picoxystrobin from cereal grains |
| CN114146565A (en) * | 2021-12-17 | 2022-03-08 | 北京腾跃高科科技有限公司 | Functional brain peptide extraction equipment and process thereof |
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