CN112725174B - Preparation device and method for biological imprinting enzyme - Google Patents
Preparation device and method for biological imprinting enzyme Download PDFInfo
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- CN112725174B CN112725174B CN202011582570.8A CN202011582570A CN112725174B CN 112725174 B CN112725174 B CN 112725174B CN 202011582570 A CN202011582570 A CN 202011582570A CN 112725174 B CN112725174 B CN 112725174B
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- C12M23/48—Holding appliances; Racks; Supports
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- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
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- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
- C12M41/18—Heat exchange systems, e.g. heat jackets or outer envelopes
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- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
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- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/40—Means for regulation, monitoring, measurement or control, e.g. flow regulation of pressure
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- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/14—Drying
Abstract
The invention discloses a preparation device and a preparation method for biological imprinting enzyme, which comprises a main reaction vessel, an inner container, a top cover and a detection assembly, the main reaction vessel is a cavity container with an opening at the top end, a placing groove is arranged on the main reaction vessel, the inner container is placed in the placing groove of the main reaction vessel, the top cover is hinged on the opening at the top end of the main reaction vessel, the invention has the advantages that, the preparation device for the biological imprinting enzyme is matched with the preparation method, and integrates the functions of temperature control, detection and the like into a whole by utilizing special equipment, therefore, the occupied space of equipment for preparing the biological imprinting enzyme is effectively saved, the practical process of the production method is fully automatically controlled, the labor cost and the time cost can be effectively saved, the preparation speed is higher, the preparation efficiency can be effectively improved, and the preparation method can meet the requirements of small-amount preparation in a laboratory and large-batch processing preparation.
Description
Technical Field
The invention relates to the technical field of biological manufacturing, in particular to a preparation device and a preparation method for a biological imprinting enzyme.
Background
The principle of the method is that the flexibility of enzyme in aqueous solution is utilized, chiral competitive inhibitor is added, then the enzyme-inhibitor compound is transferred into lipophilic solvent, so that the three-dimensional structure of the enzyme is frozen in a modified state, and the modified enzyme without the inhibitor has stereoselectivity to substrate by virtue of the memory function.
In the preparation process of the bio-imprinting enzyme, various instruments are often matched at the present stage, including but not limited to various devices such as a mixing box, a freezing box, a vacuum dryer and the like, so that the used devices are complex, the operation is complicated, and the operation process is very troublesome.
Disclosure of Invention
The invention aims to solve the problems, designs a preparation device for the bio-imprinting enzyme, and solves the problems that in the preparation process of the bio-imprinting enzyme, various instruments are mostly matched at present, including but not limited to a mixing box, a freezing box, a vacuum dryer and other various devices, the used devices are complex, the operation is complex, and the operation process is very troublesome.
The technical scheme of the invention for realizing the aim is as follows: a preparation device for biological imprinting enzyme comprises a main reaction vessel, an inner container, a top cover and a detection assembly, wherein the main reaction vessel is a cavity container with an opening at the top end, a placing groove is formed in the main reaction vessel, the inner container is placed in the placing groove of the main reaction vessel, the top cover is hinged to the opening at the top end of the main reaction vessel, the detection assembly is arranged on the top cover, the top cover is sealed with the opening at the top end of the main reaction vessel, a plurality of temperature control assemblies are annularly arranged in the main reaction vessel and positioned on the outer side of the installing groove, an auxiliary function structure is arranged on the top cover, and a filtering assembly is arranged at the bottom end of the inner container;
the auxiliary function structure includes: the device comprises an extraction groove, a one-way valve, a charging opening, an insertion groove, a mounting seat, an ultrasonic wave conduction rod, an ultrasonic generator and a limiting piece;
the top cover is provided with an extraction groove, the extraction groove is provided with a one-way valve, a charging opening is arranged on the top cover and on one side of the top cover corresponding to the extraction groove, the center of the top cover is provided with an insertion groove, the mounting seat is inserted into the insertion groove, the mounting seat is embedded with an ultrasonic wave conduction rod, the top end of the ultrasonic wave conduction rod is connected with an ultrasonic wave generator, the top end of the mounting seat horizontally extends out of an annular limiting piece, the mounting seat can be pulled out of the insertion groove so as to insert the sealing plug, the outer side of the top surface of the insertion groove matched with the mounting seat is provided with a limiting ring groove, and the limiting piece and the limiting ring groove are in interference fit;
the filter assembly includes: the filter screen, the bottom groove, the drain pipe and the control valve;
the water-saving water dispenser is characterized in that a filter screen is mounted at the bottom end of the inner container, a bottom groove is sleeved on the outer wall of the bottom end of the inner container, a water discharge pipe extends out of the bottom end of the bottom groove, a control valve is arranged on the water discharge pipe, a water discharge hole matched with the water discharge pipe is formed in the bottom end of the mounting groove, the bottom end of the inner container is opened, the bottom groove is sleeved at an opening of the bottom end of the inner container, and a sealing sleeve is arranged at the top end of the bottom groove.
The temperature control assembly includes: a plurality of temperature control grooves, a heat conducting plate and a semiconductor temperature control regulator;
the side wall of the main reaction vessel is provided with a plurality of temperature control grooves by taking the mounting groove as a central ring, the heat conducting plate is embedded in the temperature control grooves, and the heat conducting plate is connected with the semiconductor temperature control regulator.
The detection assembly comprises: the device comprises a temperature sensor, an air pressure sensor, a humidity sensor, a controller and a display screen;
install temperature sensor, baroceptor and humidity transducer on the top cap bottom surface respectively, be equipped with the controller on the top cap, the controller is connected with temperature sensor, baroceptor and humidity transducer respectively, controller one side is equipped with the display screen and is connected with the controller.
The inner bottom surface of the top cover is provided with a pair of pressing components which are pressed with the limiting plate on the top surface of the liner;
the pressing component comprises: the spring comprises a fixed seat, a slot, an insertion block, a spring and a pressing block;
the fixing seat is installed on the inner bottom surface of the top cover, a slot is formed in the center of the fixing seat, the inserting block is inserted into the slot, a spring is sleeved on the inserting block and connected with the inner side of an opening in the bottom end of the slot, and the pressing block is installed at the bottom end of the inserting block.
A placing seat is arranged below the main reaction vessel, and a cushioning fastening component is arranged in the placing seat;
the cushioning fastening assembly comprises: a plurality of damping, placement plates and fastening cushions;
the improved reaction vessel is characterized in that a placing groove is formed in the center of the placing seat, a plurality of dampers are distributed on the bottom surface of the placing groove in an annular array mode, the top ends of the dampers are connected with a placing plate, the placing plate is in contact with the bottom surface of the main reaction vessel, a corresponding groove corresponding to the drain hole is formed in one side of the placing groove, and a fastening cushion pad is arranged on the inner circle of the placing groove.
A preparation method for a biological imprinted enzyme comprises the following steps: step S1, enzyme induction, step S2, freeze drying, step S3, elution, step S4, separation;
step S1: opening the top cover of the equipment, dissolving the enzyme in a buffer solution containing a ligand of the equipment to induce the conformation of the enzyme to change, and adding the enzyme solution into the inner container of the equipment;
step S2: the inner container is placed in a mounting groove of a main reaction vessel, a top cover is buckled, a temperature control adjusting component is started to reduce the temperature in the mounting groove, the mounting groove is vacuumized, then enzyme liquid is rapidly sublimated to be freeze-dried to form enzyme powder, water is directly sublimated without melting, and in order to prevent the possible change of enzyme conformation in the freeze-drying process, a protective agent (such as inorganic salt, substrate analogue and the like) capable of maintaining the enzyme structure must be added in the process to reduce or even eliminate the influence;
step S3: adding an organic solvent into the inner container, washing the freeze-dried enzyme powder, and removing the imprinted molecules, namely an elution process;
step S4: the organic solvent washed enzyme powder was filtered and dried in vacuo.
The reaction time induced in the step S1 is set between 30min and 1 h.
And in the step S2, the temperature in the mounting groove is ensured to be between-10 and-20 ℃ during freezing.
The mesh diameter of the filter screen in the step S4 is not more than 200 meshes.
The temperature of the vacuum drying in the step S4 is set to 40 to 50 ℃.
The preparation device for the biological imprinted enzyme, which is manufactured by the technical scheme of the invention, is matched with the preparation method, and special equipment is used for integrating the functions of temperature control, detection and the like, so that the occupied space of the equipment for preparing the biological imprinted enzyme is effectively saved, meanwhile, the practical process of the production method is fully automatically controlled, the labor cost and the time cost can be effectively saved, the preparation speed is higher, the preparation efficiency can be effectively improved, the small-amount preparation in a laboratory can be met, and the large-batch processing preparation can also be met.
Drawings
FIG. 1 is a schematic sectional view of the device and method for preparing the bio-imprinted enzyme according to the present invention.
FIG. 2 is a schematic top view of the apparatus and method for preparing a bio-imprinted enzyme according to the present invention.
FIG. 3 is a schematic view of the device and method for preparing the bio-imprinted enzyme according to the present invention.
FIG. 4 is a schematic top cross-sectional view of an apparatus and a method for preparing a bio-imprinted enzyme according to the present invention.
FIG. 5 is a schematic view of a device and a method for preparing a bio-imprinted enzyme according to the present invention, showing a partially enlarged structure of FIG. 1.
In the figure: 1. a main reaction vessel; 2. an inner container; 3. a top cover; 4. drawing a groove; 5. a one-way valve; 6. a feed inlet; 7. inserting the slots; 8. a mounting seat; 9. an ultrasonic wave conduction rod; 10. an ultrasonic generator; 11. a limiting sheet; 12. a filter screen; 13. a bottom groove; 14. a drain pipe; 15. a control valve; 16. a temperature control tank; 17. a heat conducting plate; 18. a semiconductor temperature control regulator; 19. a temperature sensor; 20. an air pressure sensor; 21. a humidity sensor; 22. a controller; 23. a display screen; 24. a sealing plug; 25. sealing sleeves; 26. a fixed seat; 27. a slot; 28. inserting a block; 29. a spring; 30. briquetting; 31. a placing seat; 32. damping; 33. placing a plate; 34. the cushion pad is fastened.
Detailed Description
The invention is described in detail with reference to the accompanying drawings, and as shown in fig. 1-5, a preparation apparatus for a bio-imprinting enzyme comprises a main reaction vessel 1, a liner 2, a top cover 3 and a detection component, wherein the main reaction vessel 1 is a cavity container with an open top end, a placing groove is formed in the main reaction vessel 1, the liner 2 is placed in the placing groove of the main reaction vessel 1, the top cover 3 is hinged to the open top end of the main reaction vessel 1, the detection component is arranged on the top cover 3, the top cover 3 is sealed with the open top end of the main reaction vessel 1, a plurality of temperature control components are annularly arranged in the main reaction vessel 1 and outside the placing groove, an auxiliary functional structure is arranged on the top cover 3, and a filtering component is arranged at the bottom end of the liner 2; the auxiliary function structure includes: the device comprises an extraction tank 4, a one-way valve 5, a feed inlet 6, an insertion tank 7, a mounting seat 8, an ultrasonic wave conduction rod 9, an ultrasonic generator 10 and a limiting piece 11; an extraction groove 4 is formed in the top cover 3, a one-way valve 5 is arranged on the extraction groove 4, a charging opening 6 is formed in the top cover 3 and located on one side, corresponding to the extraction groove 4, of the top cover 3, an insertion groove 7 is formed in the center of the top cover 3, the installation seat 8 is inserted into the insertion groove 7, an ultrasonic wave conduction rod 9 is embedded in the installation seat 8, an ultrasonic wave generator 10 is connected to the top end of the ultrasonic wave conduction rod 9, an annular limiting piece 11 horizontally extends from the top end of the installation seat 8, the installation seat 8 can be pulled out of the insertion groove 7 so as to be inserted into the sealing plug 24, a limiting ring groove is formed in the outer side of the top surface of the insertion groove 7 matched with the sealing plug 24, and the limiting piece 11 and the limiting ring groove are in interference fit; the filter assembly includes: a filter screen 12, a bottom groove 13, a drain pipe 14 and a control valve 15; a filter screen 12 is installed at the bottom end of the inner container 2, a bottom groove 13 is sleeved on the outer wall of the bottom end of the inner container 2, a drain pipe 14 extends out of the bottom end of the bottom groove 13, a control valve 15 is arranged on the drain pipe 14, a drain hole matched with the drain pipe 14 is formed in the bottom end of the installation groove, the bottom end of the inner container 2 is opened, the bottom groove 13 is sleeved at an opening in the bottom end of the inner container 2, and a sealing sleeve 25 is arranged at the top end of the bottom groove 13; the temperature control assembly includes: a plurality of temperature control grooves 16, a heat conducting plate 17 and a semiconductor temperature control regulator 18; the side wall of the main reaction vessel 1 is annularly provided with a plurality of temperature control grooves 16 by taking the mounting groove as a center, the heat conducting plate 17 is embedded in the temperature control grooves 16, and the heat conducting plate 17 is connected with the semiconductor temperature control regulator 18; the detection assembly comprises: a temperature sensor 19, an air pressure sensor 20, a humidity sensor 21, a controller 22 and a display screen 23; a temperature sensor 19, an air pressure sensor 20 and a humidity sensor 21 are respectively installed on the bottom surface of the top cover 3, a controller 22 is arranged on the top cover 3, the controller 22 is respectively connected with the temperature sensor 19, the air pressure sensor 20 and the humidity sensor 21, and a display screen 23 is arranged on one side of the controller 22 and connected with the controller 22; the inner bottom surface of the top cover 3 is provided with a pair of pressing components which are pressed with the limiting plate on the top surface of the inner container 2; the pressing assembly comprises: a fixed seat 26, a slot 27, an insertion block 28, a spring 29 and a pressing block 30; the fixed seat 26 is mounted on the inner bottom surface of the top cover 3, a slot 27 is formed in the center of the fixed seat 26, the insertion block 28 is inserted into the slot 27, a spring 29 is sleeved on the insertion block 28, the spring 29 is connected with the inner side of an opening at the bottom end of the slot 27, and the pressing block 30 is mounted on the bottom end of the insertion block 28; a placing seat 31 is arranged below the main reaction vessel 1, and a cushioning fastening component is arranged in the placing seat 31; the cushioning fastening assembly comprises: a plurality of dampers 32, a placement plate 33, and a fastening cushion 34; a placing groove is formed in the center of the placing seat 31, a plurality of dampers 32 are distributed on the inner bottom surface of the placing groove in an annular array mode, a placing plate 33 is connected to the top ends of the plurality of dampers 32, the placing plate 33 is in contact with the bottom surface of the main reaction vessel 1, a corresponding groove corresponding to the drain hole is formed in one side of the placing groove, and a fastening cushion pad 34 is arranged on the inner ring of the placing groove; a preparation method for a biological imprinted enzyme comprises the following steps: step S1, enzyme induction, step S2, freeze drying, step S3, elution, step S4, separation; step S1: opening the top cover 3 of the equipment, dissolving the enzyme in a buffer solution containing a ligand of the enzyme to induce the conformation of the enzyme to change, and adding the enzyme solution into the inner container 2 of the equipment; step S2: the inner container 2 is placed in the mounting groove of the main reaction vessel 1, the top cover 3 is buckled, the temperature control adjusting component is started to reduce the temperature in the mounting groove, the mounting groove is vacuumized and then rapidly sublimated to freeze and dry enzyme liquid to form enzyme powder, water is directly sublimated without melting, and in order to prevent the possible change of enzyme conformation in the freeze-drying process, a protective agent (such as inorganic salt, substrate analogue and the like) capable of maintaining the enzyme structure is added in the process to reduce or even eliminate the influence; step S3: adding an organic solvent into the inner container 2, washing the freeze-dried enzyme powder, and removing imprinted molecules, namely an elution process; step S4: filtering and vacuum drying the enzyme powder washed by the organic solvent; the reaction time induced in the step S1 is set to be between 30min and 1 h; in the step S2, the temperature in the mounting groove is ensured to be between-10 ℃ and-20 ℃ during the freezing; the mesh diameter of the filter screen in the step S4 is not more than 200 meshes; the temperature of vacuum drying in the step S4 is set at 40 to 50 ℃.
The device is characterized by comprising a main reaction vessel 1, an inner container 2, a top cover 3 and a detection assembly, wherein the main reaction vessel 1 is a cavity container with an opening at the top end, a placing groove is formed in the main reaction vessel 1, the inner container 2 is placed in the placing groove of the main reaction vessel 1, the top cover 3 is hinged on the opening at the top end of the main reaction vessel 1, the detection assembly is arranged on the top cover 3, the top cover 3 is sealed with the opening at the top end of the main reaction vessel 1, a plurality of temperature control assemblies are annularly arranged in the main reaction vessel 1 and positioned outside the installing groove, an auxiliary function structure is arranged on the top cover 3, and a filtering assembly is arranged at the bottom end of the inner container 2; the auxiliary function structure includes: the device comprises an extraction tank 4, a one-way valve 5, a charging hole 6, an insertion tank 7, a mounting seat 8, an ultrasonic wave conduction rod 9, an ultrasonic generator 10 and a limiting piece 11; an extraction groove 4 is formed in the top cover 3, a one-way valve 5 is arranged on the extraction groove 4, a charging opening 6 is formed in the top cover 3 and located on one side, corresponding to the extraction groove 4, an insertion groove 7 is formed in the center of the top cover 3, an installation seat 8 is inserted into the insertion groove 7, an ultrasonic conduction rod 9 is embedded in the installation seat 8, an ultrasonic generator 10 is connected to the top end of the ultrasonic conduction rod 9, an annular limiting piece 11 horizontally extends out of the top end of the installation seat 8, the installation seat 8 can be pulled out of the insertion groove 7 so as to be inserted into a sealing plug 24, a limiting ring groove is formed in the outer side of the top surface of the insertion groove 7 matched with the installation seat, and the limiting piece 11 and the limiting ring groove are in interference fit; the filter assembly includes: a filter screen 12, a bottom groove 13, a drain pipe 14 and a control valve 15; the bottom end of the inner container 2 is provided with a filter screen 12, the outer wall of the bottom end of the inner container 2 is sleeved with a bottom groove 13, the bottom end of the bottom groove 13 extends out of a drain pipe 14, the drain pipe 14 is provided with a control valve 15, the bottom end of the mounting groove is provided with a drain hole matched with the drain pipe 14, the bottom end of the inner container 2 is opened, the bottom groove 13 is sleeved at an opening at the bottom end of the inner container 2, and a sealing sleeve 25 is arranged at the top end of the bottom groove 13; according to the preparation device for the biological imprinting enzyme, the preparation method is matched, and special equipment is utilized to integrate functions of temperature control, detection and the like, so that the occupied space of the equipment for preparing the biological imprinting enzyme is effectively saved, meanwhile, the practice process of the production method is fully automatically controlled, the labor cost and the time cost can be effectively saved, the preparation speed is higher, the preparation efficiency can be effectively improved, a small amount of preparation in a laboratory can be met, and the large batch of processing and preparation can also be met.
All the electrical components in the present application are connected with the power supply adapted to the electrical components through the wires, and an appropriate controller should be selected according to actual conditions to meet control requirements, and specific connection and control sequences.
Example (b): according to the attached figures 1-5 of the specification, the scheme designs a preparation device for a biological imprinting enzyme, which comprises a main reaction vessel 1, an inner container 2, a top cover 3 and a detection component, wherein the main reaction vessel 1 is a cavity container with an open top end, a placing groove is formed in the main reaction vessel 1, the inner container 2 is placed in the placing groove of the main reaction vessel 1, the top cover 3 is hinged on the open top end of the main reaction vessel 1, the detection component is arranged on the top cover 3, the top cover 3 is sealed with the open top end of the main reaction vessel 1, a plurality of temperature control components are annularly arranged in the main reaction vessel 1 and positioned outside the installing groove, an auxiliary function structure is arranged on the top cover 3, a filter component is arranged at the bottom end of the inner container 2, in the specific implementation process, the main reaction vessel 1 is used for integrally wrapping and supporting equipment, the cavity inside the main reaction vessel 1 is used for placing the inner container 2, and the inner container 2 is a reaction container for preparing the biological imprinting enzyme, meanwhile, the top cover 3 is hinged and buckled to seal the placing groove, a detection assembly is arranged at the top of the top cover 3 and used for detecting various environmental indexes in the preparation process of the biological imprinted enzyme, the placing groove is subjected to temperature control through a plurality of temperature control assemblies, and various operations in the preparation process are assisted through the cooperation of an auxiliary function structure and a filtering assembly;
as can be seen from fig. 1 to 5 of the specification, the auxiliary function structure includes: the device comprises an extraction groove 4, a one-way valve 5, a feed inlet 6, an insertion groove 7, a mounting seat 8, an ultrasonic wave conduction rod 9, an ultrasonic generator 10 and a limiting piece 11, wherein the connection relation and the position relation are as follows;
an extraction groove 4 is formed in the top cover 3, a one-way valve 5 is arranged on the extraction groove 4, a feed inlet 6 is formed in one side, corresponding to the extraction groove 4, of the top cover 3, an insertion groove 7 is formed in the center of the top cover 3, an installation seat 8 is inserted into the insertion groove 7, an ultrasonic wave conduction rod 9 is embedded in the installation seat 8, the top end of the ultrasonic wave conduction rod 9 is connected with an ultrasonic wave generator 10, an annular limiting piece 11 horizontally extends out of the top end of the installation seat 8, a limiting ring groove is formed in the outer side of the top face of the insertion groove 7 matched with the limiting piece, and the limiting piece 11 and the limiting ring groove are in interference fit;
in the specific implementation process, the top cover 3 is opened, the inner container 2 containing the reaction solution is placed on the placing groove, then the top cover 3 is buckled, buffer solution containing enzyme ligands is injected into the inner container 2 from the feed inlet 6, the mounting seat 8 is inserted into the inserting groove 7, then the ultrasonic generator 10 is started, ultrasonic waves generate vibration and are further transmitted into mixed liquid of the inner container 2 to play a role in stirring, so that the enzyme is more completely contacted with the ligands, the conformation induction of the enzyme is smoother and quicker, the mounting seat 8 can be pulled out of the inserting groove 7 and is inserted into the sealing plug 24, after the induction is finished, the limiting piece 11 and the limiting ring groove are in interference fit, the mounting seat 8 is fastened, the extraction groove 4 is connected with an external vacuum-pumping pipe, and the vacuum-pumping can be realized on the position of the placing groove when the vacuum drying is needed by controlling the one-way valve 5;
as can be seen from figures 1-5 of the specification, the filter assembly comprises: a filter screen 12, a bottom tank 13, a drain pipe 14, and a control valve 15, the connection relationship and the positional relationship of which are as follows;
a filter screen 12 is installed at the bottom end of the inner container 2, a bottom groove 13 is sleeved on the outer wall of the bottom end of the inner container 2, a drain pipe 14 extends from the bottom end of the bottom groove 13, a control valve 15 is arranged on the drain pipe 14, and a drain hole matched with the drain pipe 14 is formed in the bottom end of the installation groove;
in the specific implementation process, the bottom end of the inner container 2 is opened, the bottom groove 13 is sleeved at the opening at the bottom end of the inner container 2, the sealing sleeve 25 is arranged at the top end of the bottom groove 13, the bottom groove 13 is fixed by using the sealing sleeve 25, the drain pipe 14 is arranged below the bottom groove 13, the drain pipe 14 is provided with the control valve 15, the control valve 15 is used for controlling the opening and closing of the drain pipe 14, after freeze-drying is completed, an organic solvent is added to wash the enzyme powder, invalid components in the enzyme powder are filtered and taken out along with the enzyme powder through the filter screen 12, so that the enzyme is left, then vacuum drying is carried out, and redundant water is discharged, so that the biological imprinted enzyme with higher purity is obtained;
as can be seen from fig. 1 to 5 of the specification, the temperature control assembly includes: a plurality of temperature control grooves 16, a heat conducting plate 17 and a semiconductor temperature control regulator 18, the connection relationship and the position relationship of which are as follows;
the side wall of the main reaction vessel 1 is annularly provided with a plurality of temperature control grooves 16 by taking the mounting groove as a center, a heat conducting plate 17 is embedded in the temperature control grooves 16, and the heat conducting plate 17 is connected with a semiconductor temperature control regulator 18;
in the specific implementation process, a plurality of temperature control grooves 16 on the main reaction vessel 1 are attached to a heat conducting plate 17, the heat conducting plate 17 is connected with a semiconductor temperature control regulator 18, and the semiconductor temperature control regulator 18 is used for heating or cooling the heat conducting plate 17, so that the temperature in the placing groove is reduced or heated;
the detection assembly comprises: a temperature sensor 19, an air pressure sensor 20, a humidity sensor 21, a controller 22 and a display screen 23;
a temperature sensor 19, an air pressure sensor 20 and a humidity sensor 21 are respectively arranged on the bottom surface of the top cover 3, a controller 22 is arranged on the top cover 3, the controller 22 is respectively connected with the temperature sensor 19, the air pressure sensor 20 and the humidity sensor 21, and a display screen 23 is arranged on one side of the controller 22 and connected with the controller 22;
in the specific implementation process, the temperature in the placing groove is detected through the temperature sensor 19, the humidity in the placing groove is detected through the humidity sensor 21, the air pressure in the placing groove is detected through the air pressure detector, the sensors respectively gather data into the controller 22, and the data are processed and displayed on the display screen 23 through the controller 22 for a producer to observe and adjust;
as can be seen from fig. 1-5 of the specification, a pair of pressing components are arranged on the inner bottom surface of the top cover 3 and are pressed with the limiting plate on the top surface of the liner 2; the lamination assembly comprises: the fixing seat 26, the slot 27, the insertion block 28, the spring 29 and the pressing block 30 are connected and positioned as follows;
the fixed seat 26 is arranged on the inner bottom surface of the top cover 3, a slot 27 is formed in the center of the fixed seat 26, the insertion block 28 is inserted into the slot 27, a spring 29 is sleeved on the insertion block 28, the spring 29 is connected with the inner side of an opening at the bottom end of the slot 27, and the pressing block 30 is arranged on the bottom end of the insertion block 28;
in the specific implementation process, the slot 27 on the fixed seat 26 is matched with the insert block 28, the insert block 28 can stretch out and draw back in the slot 27, the elasticity of the spring 29 is utilized, when the top cover 3 is buckled, the fixed seat 26, the insert block 28 and the pressing block 30 are driven to press downwards, when the pressing block 30 contacts the limiting sheet 11, the insert block 28 is pushed to slide upwards in the slot 27, so that the spring 29 is stretched, the stretched tensile force is used as the acting force of the top surface of the limiting liner 2, and the liner 2 is prevented from shaking;
as can be seen from fig. 1-5 of the specification, a placing seat 31 is arranged below the main reaction vessel 1, and a cushioning fastening component is arranged in the placing seat 31; the bradyseism fastening component includes: a plurality of dampers 32, a placing plate 33 and a fastening cushion 34, the connection and position relationships of which are as follows;
a placing groove is formed in the center of the placing seat 31, a plurality of dampers 32 are distributed on the inner bottom surface of the placing groove in an annular array mode, the top ends of the plurality of dampers 32 are connected with a placing plate 33, the placing plate 33 is in contact with the bottom surface of the main reaction vessel 1, a corresponding groove corresponding to the drain hole is formed in one side of the placing groove, and a fastening cushion pad 34 is arranged on the inner ring of the placing groove;
in the concrete implementation process, settle main reaction ware 1 in the standing groove of placing seat 31 during the use, align wash port and corresponding groove, utilize and place board 33 and bear main reaction ware 1, carry out the bottom shock attenuation to main reaction ware 1 through damping 32, utilize fastening buffer pad 34 ring to locate the main reaction ware 1 outside, provide horizontally fixed effect, avoid in the use, the condition that main reaction ware 1 rocked appears.
The preparation method of the biological imprinted enzyme based on the equipment comprises the following steps: firstly, opening a top cover 3 of the equipment, dissolving enzyme in a buffer solution containing a ligand of the equipment to induce the conformation of the enzyme to change, adding the enzyme solution into an inner container 2 of the equipment, and setting the reaction time of induction between 30min and 1 h; secondly, the inner container 2 is placed in the mounting groove of the main reaction vessel 1, the top cover 3 is buckled, the temperature control adjusting component is started to reduce the temperature in the mounting groove, the mounting groove is vacuumized, then the temperature is rapidly sublimated to freeze and dry enzyme liquid to form enzyme powder, the water is directly sublimated without melting, the conformation of the enzyme can be fixed, in order to prevent the possible change of the conformation of the enzyme in the freeze-drying process, a protective agent (such as inorganic salt, substrate analogs and the like) capable of maintaining the enzyme structure is added in the process to reduce or even eliminate the influence, and the temperature in the mounting groove is ensured to be between-10 ℃ and-20 ℃ during freezing; thirdly, the method comprises the following steps: adding an organic solvent into the inner container 2, washing the freeze-dried enzyme powder, and removing imprinted molecules, namely an elution process; fourthly, the method comprises the following steps: filtering enzyme powder washed by organic solvent and vacuum drying, wherein the mesh diameter of the filter screen 12 is not more than 200 meshes, and the vacuum drying temperature is set at 40-50 ℃;
in conclusion, the preparation device for the bio-imprinting enzyme is matched with the preparation method, and the functions of temperature control, detection and the like are integrated into a whole by using special equipment, so that the occupied space of the equipment for preparing the bio-imprinting enzyme is effectively saved, meanwhile, the practical process of the production method is fully automatically controlled, the labor cost and the time cost can be effectively saved, the preparation speed is higher, the preparation efficiency can be effectively improved, and the preparation device can meet the requirements of small-amount preparation in a laboratory and large-batch processing preparation.
The technical solutions described above only represent the preferred technical solutions of the present invention, and some possible modifications to some parts of the technical solutions by those skilled in the art all represent the principles of the present invention, and fall within the protection scope of the present invention.
Claims (10)
1. A preparation device for biological imprinting enzyme comprises a main reaction vessel (1), an inner container (2), a top cover (3) and a detection component, and is characterized in that the main reaction vessel (1) is a cavity container with an open top end, a placing groove is formed in the main reaction vessel (1), the inner container (2) is placed in the placing groove of the main reaction vessel (1), the top cover (3) is hinged to the open top end of the main reaction vessel (1), the detection component is arranged on the top cover (3), the top cover (3) is sealed with the open top end of the main reaction vessel (1), a plurality of temperature control components are annularly arranged in the main reaction vessel (1) and positioned outside the installing groove, an auxiliary function structure is arranged on the top cover (3), and a filtering component is arranged at the bottom end of the inner container (2);
the auxiliary function structure includes: the device comprises an extraction groove (4), a one-way valve (5), a feed inlet (6), an insertion groove (7), a mounting seat (8), an ultrasonic wave conduction rod (9), an ultrasonic wave generator (10) and a limiting piece (11);
the sealing plug is characterized in that an extraction groove (4) is formed in the top cover (3), a check valve (5) is arranged on the extraction groove (4), a feed opening (6) is formed in one side, corresponding to the extraction groove (4), of the top cover (3), an insertion groove (7) is formed in the center of the top cover (3), an installation seat (8) is inserted into the insertion groove (7), an ultrasonic conduction rod (9) is embedded into the installation seat (8), an ultrasonic generator (10) is connected to the top end of the ultrasonic conduction rod (9), an annular limiting piece (11) horizontally extends out of the top end of the installation seat (8), the installation seat (8) can be pulled out of the insertion groove (7), so that a sealing plug (24) is inserted, a limiting ring groove is formed in the outer side of the top surface of the insertion groove (7) matched with the sealing plug, and the limiting piece (11) and the limiting ring groove are in interference fit;
the filter assembly includes: a filter screen (12), a bottom groove (13), a drain pipe (14) and a control valve (15);
filter screen (12) are installed to inner bag (2) bottom, inner bag (2) bottom outer wall cover is equipped with kerve (13), drain pipe (14) are extended to kerve (13) bottom, be equipped with control valve (15) on drain pipe (14), the mounting groove bottom is equipped with the wash port that matches with drain pipe (14), inner bag (2) bottom is opened, inner bag (2) bottom opening part is located to kerve (13) cover, and is located kerve (13) top and is equipped with seal cover (25).
2. The apparatus of claim 1, wherein the temperature control assembly comprises: a plurality of temperature control grooves (16), a heat conducting plate (17) and a semiconductor temperature control regulator (18);
the side wall of the main reaction vessel (1) is provided with a plurality of temperature control grooves (16) by taking the mounting groove as a central ring, the heat conducting plate (17) is embedded in the temperature control grooves (16), and the heat conducting plate (17) is connected with the semiconductor temperature control regulator (18).
3. The manufacturing apparatus of claim 1, wherein the detection component comprises: a temperature sensor (19), an air pressure sensor (20), a humidity sensor (21), a controller (22) and a display screen (23);
install temperature sensor (19), baroceptor (20) and humidity transducer (21) on top cap (3) bottom surface respectively, be equipped with controller (22) on top cap (3), controller (22) are connected with temperature sensor (19), baroceptor (20) and humidity transducer (21) respectively, controller (22) one side is equipped with display screen (23) and is connected with controller (22).
4. The preparation device for the bio-imprinting enzyme according to claim 1, wherein a pair of pressing components are arranged on the inner bottom surface of the top cover (3) and are pressed with the limiting plate on the top surface of the inner container (2);
the pressing component comprises: a fixed seat (26), a slot (27), an insertion block (28), a spring (29) and a pressing block (30);
the fixing seat (26) is installed on the bottom surface in the top cover (3), a slot (27) is formed in the center of the fixing seat (26), an insertion block (28) is inserted into the slot (27), a spring (29) is sleeved on the insertion block (28), the spring (29) is connected with the inner side of an opening in the bottom end of the slot (27), and a pressing block (30) is installed on the bottom end of the insertion block (28).
5. The preparation device for the bio-imprinting enzyme according to claim 1, wherein a placing seat (31) is arranged below the main reaction vessel (1), and a shock absorption fastening component is arranged in the placing seat (31);
the cushioning fastening assembly comprises: a plurality of dampers (32), a placement plate (33) and a fastening cushion (34);
the novel wash basin is characterized in that a placing groove is formed in the center of the placing seat (31), a plurality of dampers (32) are distributed on the bottom surface of the placing groove in an annular array mode, the top ends of the dampers (32) are connected with a placing plate (33), the placing plate (33) is in contact with the bottom surface of the main reaction vessel (1), a corresponding groove corresponding to the drain hole is formed in one side of the placing groove, and a fastening buffer pad (34) is arranged on the inner circle of the placing groove.
6. A method for producing a bio-blotting enzyme using the apparatus for producing a bio-blotting enzyme according to any of claims 1 to 5, comprising the steps of: step S1, enzyme induction, step S2, freeze drying, step S3, elution, step S4 and separation;
step S1: opening the top cover (3) of the device, dissolving the enzyme in a buffer solution containing a ligand of the enzyme, inducing the conformation of the enzyme to change, and adding the enzyme solution into the inner container (2) of the equipment;
step S2: the inner container (2) is placed in the mounting groove of the main reaction vessel (1), the top cover (3) is buckled, the temperature control component is started to reduce the temperature in the mounting groove, the mounting groove is vacuumized, then the temperature control component is quickly sublimated to freeze and dry enzyme liquid to form enzyme powder, water is directly sublimated without melting, the conformation of the enzyme can be fixed, and in order to prevent the possible change of the conformation of the enzyme in the freeze-drying process, a protective agent capable of maintaining the structure of the enzyme must be added in the process to reduce or even eliminate the influence;
step S3: adding an organic solvent into the inner container (2), washing the freeze-dried enzyme powder, and removing the imprinted molecules, namely the elution process;
step S4: the organic solvent washed enzyme powder was filtered and dried in vacuo.
7. The method for preparing a bio-imprinted enzyme according to claim 6, wherein the reaction time induced in the step S1 is set between 30min and 1 h.
8. The method for preparing a bio-imprinted enzyme according to claim 6, wherein the temperature in the mounting groove is maintained between-10 to-20 ℃ during the freezing in step S2.
9. The method of claim 6, wherein the mesh diameter of the filter in step S4 is not larger than 200 mesh.
10. The method for preparing a bio-imprinted enzyme according to claim 6, wherein the temperature of vacuum drying in step S4 is set to 40-50 ℃.
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