CN114292837B - Immobilization method for producing enzyme special for 4-aminobutyric acid and oscillation device thereof - Google Patents

Abstract

The invention discloses an immobilization method of special enzyme for producing 4-aminobutyric acid and an oscillation device thereof, relating to the technical field of enzyme immobilization, comprising the following steps of S1 resin pretreatment; s2 glutaraldehyde crosslinking; the S3 enzyme is fixed, and the device comprises the container, a positioning and supporting mechanism for positioning the container and a driving unit for driving the container to move; the positioning support mechanism is provided with a strip-shaped offset channel which is parallel to the horizontal plane and has the same width, and a sliding piece which is assembled in the strip-shaped offset channel and can slide along the length direction of the strip-shaped offset channel, wherein a circular through hole for a container to pass through is formed in the middle position of the sliding piece; the container is fitted inside the circular through-hole through the circular through-hole, and the container is axially rotatable in the circular through-hole. When the enzyme immobilization method is applied to industrial production in the field of biocatalysis, the enzyme immobilization method can be used for immobilization of enzymes in a large scale through single preparation, and the production and preparation efficiency is high.

Description

Immobilization method for producing enzyme special for 4-aminobutyric acid and oscillation device thereof

Technical Field

The invention relates to the technical field of enzyme immobilization, in particular to an immobilization method for producing special enzyme for 4-aminobutyric acid and an oscillation device thereof.

Background

Enzymes immobilized on solid materials are a major form of biocatalyst application due to the advantages of easy catalyst recovery, more flexible reactor design and relatively simple product purification. Enzymes have been immobilized on various inorganic or organic solid supports by various chemical or physical methods, or embedded in hollow fibers or microcapsules, or crosslinked by covalent bonds. The structure and surface properties of the support material, as well as the manner in which the enzyme is bound to the material, are among many factors critical to the performance of the immobilized enzyme. Researchers have been working to optimize the structure of the support material to produce more efficient biocatalysts.

The immobilization methods of enzymes can be divided into four main categories: adsorption, entrapment, covalent bonding, and cross-linking. Adsorption is the earliest enzyme immobilization method that occurs, including physical adsorption and ion exchange adsorption. The method has mild conditions, and the conformational change of the enzyme is small or basically unchanged, so that the method has small influence on the catalytic activity of the enzyme, but the bonding force between the enzyme and the carrier is weak, and under the conditions of uncomfortable pH, high salt concentration, high temperature and the like, the enzyme is easy to fall off from the carrier and pollute catalytic reaction products and the like. The basic principle of the embedding method is that after the carrier is mixed with the enzyme solution, polymerization reaction is carried out by virtue of an initiator, and the enzyme is limited in the grids of the carrier through physical action, so that the enzyme immobilization method is realized. The method does not involve the conformation of the enzyme and the chemical change of the enzyme molecules, and has mild reaction conditions, so that the recovery rate of the enzyme activity is higher. The immobilized enzyme of the embedding method is easy to leak, has the problems of diffusion limitation and the like, and is not suitable for catalyzing the reaction of macromolecular substrates because the catalytic reaction is influenced by mass transfer resistance. The covalent bonding method is an enzyme immobilization method for realizing irreversible bonding by forming chemical covalent bonds between an unnecessary group of an enzyme molecule and an active functional group on the surface of a carrier, and the obtained immobilized enzyme is firmly connected with the carrier and has good stability and reusability, but the method has stronger reaction and more serious activity loss of the immobilized enzyme than other immobilization methods. The crosslinking method is to use bifunctional or multifunctional crosslinking reagent to form covalent bond between enzyme molecule and crosslinking reagent, and to use different crosslinking condition and add different materials into crosslinking system to generate immobilized enzyme with different physical properties. The crosslinking method is generally used as an aid to other immobilization methods. The method has the advantages of more available crosslinking reagents and simple technology; the binding force of the enzyme is strong and very stable. However, the above method has a disadvantage of low production efficiency in industrial production applied to the field of biocatalysis.

Disclosure of Invention

The invention aims to provide an oscillation device for producing enzyme immobilization special for 4-aminobutyric acid, which solves the defect of low preparation efficiency in industrial production applied to the field of biocatalysis in the prior art.

In order to achieve the aim of the invention, the invention adopts the following technical scheme:

the invention provides an immobilization method for producing 4-aminobutyric acid special enzyme, which comprises the following steps of;

Pretreatment of S1 resin: weighing a proper amount of the amination carrier, pouring the amination carrier into a container of a shaking device, adding a certain amount of 0.1mol/L PBS buffer solution, shaking at a temperature of 60 ℃ and a rotating speed of 100rpm for 15 minutes, standing at room temperature for 1 hour, taking out the amination carrier, filtering with filter paper, pouring the amination carrier into the container 1 of the shaking device again, and adding a proper amount of 0.02mol/L PBS buffer solution;

S2 glutaraldehyde crosslinking: adding glutaraldehyde solution with the concentration of 50% to reach the final concentration of 2%, oscillating for 5 hours at the temperature of 25 ℃ and the rotating speed of 60rpm, taking out the crosslinked product, and washing 3 times with distilled water;

Immobilization of S3 enzyme: pouring into a container of an oscillating device again, immobilizing by adopting TGE buffer solution with PH=7.9, weighing a proper amount of pretreated resin, adding 1.5ml of corresponding buffer solution and 0.5ml of crude enzyme solution, oscillating and fixing for 10 hours at the temperature of 16 ℃ and the rotating speed of 50rpm, taking out a solidified product, washing for 3 times by using the buffer solution, finally adding a small amount of the buffer solution to immerse the immobilized enzyme, adding a stabilizing agent, and preserving at normal temperature.

Further, the stabilizing agent is enzyme stabilizing agent SG03.

An oscillation device of an immobilization method of an enzyme special for producing aminobutyric acid, comprising a container, a positioning and supporting mechanism for positioning the container and a driving unit for driving the container to move;

the positioning support mechanism is provided with a strip-shaped offset channel which is parallel to the horizontal plane and has the same width, and a sliding piece which is assembled in the strip-shaped offset channel and can slide along the length direction of the strip-shaped offset channel, wherein a circular through hole for a container to pass through is formed in the middle position of the sliding piece;

the container is assembled inside the circular through hole through the circular through hole, and the container can axially rotate in the circular through hole;

the driving unit comprises a first driving mechanism for driving the container to reciprocate along the length direction of the strip-shaped offset channel and a second driving mechanism for driving the container to synchronously rotate at a non-uniform speed.

Further, the location supporting mechanism includes the location base plate that the level set up, strip offset passageway is offered on the location base plate, the slider has the conflict limit of laminating with strip offset passageway both sides wall, and the both ends are close to strip offset passageway both sides about the conflict limit and all extend outwards and be formed with the chimb, chimb relative location base plate one side evenly is provided with the arc recess, and the inside card of arc recess is equipped with the spin.

Further, the container is cylindric wholly, and the inside of container is provided with the hybrid chamber, the top of container is provided with the feed inlet, and feed inlet position department still is provided with the lid, the axle center position department of terminal surface all is provided with the fixed column perpendicularly about the container, location supporting mechanism still includes the U-shaped locating plate, and the both ends of U-shaped locating plate are provided with the spacing through-hole of strip skew passageway looks adaptation and cover establish in the strip outside two fixed columns, wherein, U-shaped locating plate and positioning substrate fixed connection, the container bottom runs through downwards along its axle center and is provided with the discharge gate, and is provided with the valve in discharge gate department.

Further, the first driving mechanism comprises lever components symmetrically arranged on the upper end face and the lower end face of the U-shaped positioning plate, a connecting component used for connecting the lever components and driving the lever components to swing and a driving source; wherein the lever assembly comprises;

The support column is fixed at one end of the U-shaped positioning plate, which is close to the strip-shaped limiting through hole; and

The swing rod is rotationally connected with the support column, and axially rotates along the support column, and a through hole for the fixing column to pass through is formed in one end, close to the fixing column, of the other end of the swing rod;

The connection assembly includes:

The two ends of the fixed rod are fixedly connected with the other ends of the two swing rods respectively;

The deflection block is fixed in the middle of the fixed rod, and a strip through groove is formed in the deflection block in a penetrating manner; and

The two support plates are symmetrically arranged on the side walls of the U-shaped positioning plates at two sides of the deflection block, the opposite sides of the two support plates are coaxially and rotatably provided with rotating disks, and a force application rod penetrating through the strip-shaped through groove is fixed at the edge position between the two rotating disks;

the driving source is used for driving the rotating disc to circumferentially rotate, the driving source is a speed reducer and a motor, the input end of the speed reducer is fixedly connected with the output end of the motor, and the output end of the speed reducer is fixedly connected with the driving shaft of the rotating disc;

the strip-shaped offset channels are arc-shaped, and the circle centers of the strip-shaped offset channels are positioned on the central axis of the support column.

Further, the second driving mechanism comprises a one-way bearing assembled on the fixed column, a first gear assembled outside the one-way bearing and a second gear which is fixed outside the support column and is meshed with the first gear mutually.

Further, an outer ring barrel used for encircling the positioning substrate and the driving unit is arranged on the outer side of the positioning substrate, the outer ring barrel is fixedly connected with the positioning substrate, and support legs are uniformly arranged at the bottom of the outer ring barrel.

Further, the buffer mechanism comprises a piston cylinder, the piston cylinder is fixed on the inner wall of the outer ring cylinder, a piston is slidably arranged in the piston cylinder, a piston rod is arranged on one side of the piston, one end of the piston rod extends to one side of the piston cylinder, a push plate is arranged on the other side of the piston, which is far away from the piston rod, of the piston cylinder, a return spring is arranged on the other side, which is far away from the piston rod, an air inlet pipe and an air outlet pipe are respectively arranged at one end, which is far away from the piston rod, of the piston cylinder, and a first one-way valve and a second one-way valve are respectively arranged on the air outlet pipe and the air inlet pipe;

The buffer mechanism further comprises a baffle plate arranged on the outer side of the container and a T-shaped air ejector tube coaxially arranged inside the container, wherein the top end of the T-shaped air ejector tube penetrates through the top of the container and a fixing column positioned at the top of the T-shaped air ejector tube is connected with the air outlet tube through a hose, and when the container moves, the baffle plate can push the push plate in a clearance mode to enable the piston to generate displacement.

Further, the heating device comprises a heating mechanism, wherein the heating mechanism comprises a heating pipe fixed inside the container, a temperature sensor for detecting the temperature inside the container and a controller.

Compared with the prior art, the above technical scheme has the following beneficial effects:

The immobilized enzyme is obtained by immobilizing the enzyme by a crosslinking method, and chemically connecting glutaraldehyde with enzyme crosslinking, so that the enzyme obtains hydrophobicity and can be separated from a solution.

When the oscillation device provided by the invention is applied to industrial production in the field of biocatalysis, the enzyme is immobilized on a large scale by single preparation, and the production and preparation efficiency is high.

Drawings

The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention. In the drawings:

FIG. 1 is a schematic illustration of the overall structure provided by one or more embodiments of the invention;

FIG. 2 is a schematic illustration of one or more embodiments of the present invention with an outer ring cartridge integrally removed;

FIG. 3 is a schematic top view of one or more embodiments of the present invention;

FIG. 4 is a schematic view of the A-A plane structure of FIG. 3;

FIG. 5 is a schematic view of the B-B surface structure of FIG. 3;

FIG. 6 is a schematic view of the partial structure at A of FIG. 3;

FIG. 7 is a schematic view of the partial structure at B of FIG. 3;

FIG. 8 is a schematic diagram of a buffer mechanism according to one or more embodiments of the present invention.

In the figure:

1. A container; 2. positioning and supporting mechanisms; 3. a driving unit; 4. an outer ring barrel; 5. a buffer mechanism;

11. a feed inlet; 12. fixing the column; 13. a discharge port;

21. a strip-shaped offset channel; 22. a slider; 23. positioning a substrate; 24. a U-shaped positioning plate;

31. A first driving mechanism; 32. a second driving mechanism;

51. A piston cylinder; 52. a piston; 53. a piston rod; 54. a push plate; 55. a return spring; 57. an air outlet pipe; 58. a first one-way valve; 510. a baffle; 511. a T-shaped gas jet pipe;

221. A circular through hole; 222. a collision edge; 223. a convex edge; 224. an arc-shaped groove; 225. a rolling ball;

241. a strip-shaped limiting through hole;

311. a lever assembly; 312. a connection assembly; 313. a driving source;

321. A one-way bearing; 322. a first gear; 323. a second gear;

3111. A support column; 3112. swing rod; 3113. a through hole;

3121. a fixed rod; 3122. a deflection block; 3123. a strip-shaped through groove; 3124. a support plate; 3125. a rotating disc; 3126. and a force application rod.

Detailed Description

In order that those skilled in the art will better understand the present application, a technical solution in the embodiments of the present application will be clearly and completely described below with reference to the accompanying drawings in which it is apparent that the described embodiments are only some embodiments of the present application, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the present application without making any inventive effort, shall fall within the scope of the present application.

It should be noted that the terms "first," "second," and the like in the description and the claims of the present application and the above figures are used for distinguishing between similar objects and not necessarily for describing a particular sequential or chronological order. It is to be understood that the data so used may be interchanged where appropriate in order to describe the embodiments of the application herein. Furthermore, the terms "comprises," "comprising," and "having," and any variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, system, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed but may include other steps or elements not expressly listed or inherent to such process, method, article, or apparatus.

In the present application, the terms "upper", "lower", "left", "right", "front", "rear", "top", "bottom", "inner", "outer", "middle", "vertical", "horizontal", "lateral", "longitudinal" and the like indicate an azimuth or a positional relationship based on that shown in the drawings. These terms are only used to better describe the present application and its embodiments and are not intended to limit the scope of the indicated devices, elements or components to the particular orientations or to configure and operate in the particular orientations.

Also, some of the terms described above may be used to indicate other meanings in addition to orientation or positional relationships, for example, the term "upper" may also be used to indicate some sort of attachment or connection in some cases. The specific meaning of these terms in the present application will be understood by those of ordinary skill in the art according to the specific circumstances.

Furthermore, the terms "mounted," "configured," "provided," "connected," "coupled," and "sleeved" are to be construed broadly. For example, it may be a fixed connection, a removable connection, or a unitary construction; may be a mechanical connection, or an electrical connection; may be directly connected, or indirectly connected through intervening media, or may be in internal communication between two devices, elements, or components. The specific meaning of the above terms in the present application can be understood by those of ordinary skill in the art according to the specific circumstances.

As shown in FIGS. 1 to 8, a method for immobilizing an enzyme dedicated to the production of 4-aminobutyric acid comprises the steps of;

Pretreatment of S1 resin: weighing a proper amount of the amination carrier, pouring the amination carrier into a container 1 of a shaking device, adding a certain amount of 0.1mol/L PBS buffer solution, shaking at a temperature of 60 ℃ and a rotating speed of 100rpm for 15 minutes, standing at room temperature for 1 hour, taking out the amination carrier, filtering with filter paper, pouring the amination carrier into the container 1 of the shaking device again, and adding a proper amount of 0.02mol/L PBS buffer solution;

S2 glutaraldehyde crosslinking: adding glutaraldehyde solution with the concentration of 50% to reach the final concentration of 2%, oscillating for 5 hours at the temperature of 25 ℃ and the rotating speed of 60rpm, taking out the crosslinked product, and washing 3 times with distilled water;

Immobilization of S3 enzyme: pouring into a container 1 of an oscillating device again, immobilizing by adopting TGE buffer with PH=7.9, weighing a proper amount of pretreated resin, adding 1.5ml of corresponding buffer and 0.5ml of crude enzyme solution, oscillating and fixing for 10 hours at the temperature of 16 ℃ and the rotating speed of 50rpm, taking out a solidified product, washing for 3 times by using the buffer, finally adding a small amount of enzyme immersed and fixed by the buffer, adding a stabilizing agent, and preserving at normal temperature.

In this example, the stabilizing agent is enzyme stabilizing agent SG03.

The invention also provides an oscillating device of the immobilization method for producing the 4-aminobutyric acid special enzyme, which comprises a container 1, a positioning and supporting mechanism 2 for positioning the container 1 and a driving unit 3 for driving the container 1 to move;

the positioning support mechanism 2 is provided with a strip-shaped offset channel 21 parallel to the horizontal plane and having the same width, and a sliding piece 22 which is assembled inside the strip-shaped offset channel 21 and can slide along the length direction of the strip-shaped offset channel 21, wherein a circular through hole 221 for the container 1 to pass through is formed in the middle position of the sliding piece 22;

The container 1 is fitted inside the circular through-hole 221 through the circular through-hole 221, and the container 1 can axially rotate inside the circular through-hole 221;

The driving unit 3 includes a first driving mechanism 31 for driving the container 1 to reciprocate along the length direction of the strip-shaped offset passage 21 and a second driving mechanism 32 for driving the container 1 to rotate at a non-uniform speed in synchronization.

In application, the first driving mechanism 31 drives the container 1 to reciprocate along the length direction of the strip-shaped offset channel 21 to generate vibration, and meanwhile, the second driving mechanism 32 drives the container to synchronously rotate at a non-uniform speed, so that compared with the existing oscillation mode, the existing oscillation mode has fixed vibration direction, and the movement of a solution at the position close to the vibration source is larger than that of other positions, so that the mixing efficiency of all parts is different, namely, too much enzyme is solidified on a carrier at the position close to the vibration source, and too little enzyme is solidified on the carrier at other positions, thereby affecting the enzyme fixing effect.

The positioning support mechanism 2 comprises a positioning substrate 23 which is horizontally arranged, the strip-shaped offset channel 21 is arranged on the positioning substrate 23, the sliding piece 22 is provided with a collision edge 222 which is attached to two side walls of the strip-shaped offset channel 21, the upper end and the lower end of the collision edge 222 are close to two sides of the strip-shaped offset channel 21 and extend outwards to form a convex edge 223, one side of the convex edge 223 opposite to the positioning substrate 23 is uniformly provided with an arc-shaped groove 224, and a rolling ball 225 is arranged in the arc-shaped groove 224 in a clamping mode. The abutting edges 222 on two sides of the sliding piece 22 abut against two sides of the strip-shaped offset channel 21 to ensure that the sliding piece 22 can slide along the length direction of the strip-shaped offset channel 21, stability during sliding is improved, the convex edge 223 and the rolling ball 225 are arranged, the convex edge 223 can vertically limit the sliding piece 22, the sliding piece 22 is prevented from vertically shaking during sliding, and the rolling ball 225 can reduce friction of the sliding piece 22 during moving, so that load of a driving piece is reduced.

The whole cylindric that is of container 1, and the inside of container 1 is provided with the mixing chamber, the top of container 1 is provided with feed inlet 11, and feed inlet 11 position department still is provided with the lid, the axle center position department of terminal surface all is provided with fixed column 12 perpendicularly about the container 1, location supporting mechanism 2 still includes U-shaped locating plate 24, the both ends of U-shaped locating plate 24 are provided with strip offset passageway 21 looks adaptation and cover establish the spacing through-hole 241 in the strip in two fixed column 12 outsides, wherein, U-shaped locating plate 24 and positioning substrate 23 fixed connection, container 1 bottom runs through downwards along its axle center and is provided with discharge gate 13, and be provided with the valve in discharge gate 13 department. The U-shaped locating plate 24 limits the top and the bottom of the container 1, so that the stability of the container 1 during movement is further improved.

After the method of the present invention is applied to the industrial production, if only a certain amount of enzyme solution is immobilized at a time, the production efficiency must not conform to the industrial production, and if the enzyme solution is immobilized on a large scale, that is, a certain amount of immobilized enzyme must be reached at a time, the overall weight of the container increases during the oscillation, the load of the driving unit 3 increases, and the vibration frequency of the container is affected, taking this problem into consideration.

The first driving mechanism 31 comprises a lever assembly 311 symmetrically arranged on the upper end surface and the lower end surface of the U-shaped positioning plate 24, a connecting assembly 312 for connecting the lever assembly 311 and driving the lever assembly 311 to swing, and a driving source 313; wherein the lever assembly 311 comprises; the support column 3111 is fixed at one end of the U-shaped positioning plate 24 close to the strip-shaped limiting through hole 241; and a swing link 3112 rotatably connected to the support column 3111, the swing link 3112 rotating axially along the support column 3111, the other end of the swing link 3112 being provided with a through hole 3113 for the fixed column 12 to pass through near one end of the fixed column 12; the connection assembly 312 includes: the two ends of the fixed rod 3121 which is vertically arranged are fixedly connected with the other ends of the two swing rods 3112 respectively; a deflection block 3122 fixed in the middle of the fixed rod 3121, and a strip-shaped through slot 3123 is arranged on the deflection block 3122 in a penetrating manner; the two support plates 3124 are symmetrically arranged on the side walls of the U-shaped positioning plates 24 at the two sides of the deflection block 3122, the opposite sides of the two support plates 3124 are coaxially provided with rotating disks 3125 in a rotating way, and a force application rod 3126 passing through the strip-shaped through groove 3123 is fixed at the edge position between the two rotating disks 3125;

The driving source 313 is used for driving the rotating disc 3125 to rotate circumferentially, the driving source 313 is a speed reducer and a motor, an input end of the speed reducer is fixedly connected with an output end of the motor, and an output end of the speed reducer is fixedly connected with a driving shaft of the rotating disc 3125.

The invention utilizes the lever principle to drive the rotating disk 3125 to rotate by the driving source, so that the force application rod 3126 moves circularly around the axis of the rotating disk 3125, because the force application rod 3126 is inserted in the strip through groove 3123 of the deflection block 3122, when the force application rod 3126 moves, the fixed rod 3121 can swing, and then the other end of the swing rod 3112 is driven to swing, because the fulcrum-support column 3111 of the swing rod 3112 is arranged near one end close to the strip limiting through hole 241, namely the fixed rod 3121 drives the long arm end of the swing rod 3112 to swing, the short arm end of the swing rod 3112 drives the fixed column 12 to move, thereby the container 1 can swing to generate vibration, the load of the driving mechanism one is reduced, and the container 1 can keep higher vibration frequency, thus the production efficiency is improved.

Correspondingly, the strip-shaped offset channel 21 is arc-shaped, and the center of the strip-shaped offset channel 21 is on the central axis of the support column 3111.

The second driving mechanism 32 includes a one-way bearing 321 fitted on the fixed post 12, a first gear 322 fitted outside the one-way bearing 321, and a second gear 323 fixed outside the support post 3111 and intermeshed with the first gear 322. In the process of driving the fixed column 12 to swing bidirectionally by the swing rod 3112, when the fixed column 12 swings to one side, the unidirectional bearing is in a locking state, at this time, the first gear 322 is fixed on the fixed column 12, the first gear 322 is meshed with the second gear 323, so that the fixed column 12 rotates to drive the container 1 to rotate, and when the fixed column 12 swings to the other side, the unidirectional bearing is in an active state, at this time, the first gear 322 is fixed on the fixed column 12 through the bearing, the first gear 322 reversely rotates, the container 1 still keeps rotating forward, the rotating direction of the container 1 cannot be changed, and it is noted that when the rotating speed of the container 1 reaches a certain value, no rotating force is applied to the container 1 even if the fixed column 12 swings to one side, so that the container 1 rotates at a non-uniform speed, on one hand, the solution inside the container 1 can be more violently, and on the other hand, each point of the container can move to the vibration source position.

The outside of the positioning substrate 23 is provided with an outer ring barrel 4 for encircling the positioning substrate 23 and the driving unit 3, the outer ring barrel 4 is fixedly connected with the positioning substrate 23, and the bottom of the outer ring barrel 4 is uniformly provided with supporting legs. The container 1 is protected from the operator touching the container 1 during operation.

The weight of the container 1 is considered in order to prevent the rotational speed thereof from being excessively large.

The buffer mechanism 5 is used for slowing down the rotation of the container 1, the buffer mechanism 5 comprises a piston cylinder 51, the piston cylinder 51 is fixed on the inner wall of the outer ring cylinder 4, a piston 52 is slidably arranged in the piston cylinder 51, a piston rod 53 is arranged on one side of the piston 52, a push plate 54 is arranged on one side of the piston rod 53 extending to the piston cylinder 51, a reset spring 55 is arranged on the other side of the piston 52 away from the piston rod 53, an air inlet pipe 56 and an air outlet pipe 57 are respectively arranged on one end of the piston cylinder 51 away from the piston rod 53, and a first one-way valve 58 and a second one-way valve are respectively arranged on the air outlet pipe 57 and the air inlet pipe; the buffer mechanism 5 further comprises a baffle 510 arranged on the outer side of the container 1 and a T-shaped air jet pipe 511 coaxially arranged in the container 1, wherein the top end of the T-shaped air jet pipe 511 penetrates through the top of the container 1, a fixed column 12 positioned on the top is connected with the air outlet pipe 57 through a hose, and when the container 1 moves, the baffle 510 can push the push plate 54 to enable the piston 52 to displace in a clearance mode. When the container 1 moves, the baffle 510 and the push plate 54 can push the push plate 54 to enable the piston 52 to displace and compress the return spring 55, the return spring 55 can slow down the rotation speed of the container 1, meanwhile, the piston 52 moves to guide air of the piston cylinder 51 into the container 1, and the air is discharged from the container near the vibration source, and the liquid at the vibration source moves vigorously, so that bubbles in the liquid can be broken, and the generated energy can be beneficial to the more permeable carrier porosity and the enzyme load rate.

The heating device comprises a container 1, a heating mechanism, a temperature sensor and a controller, wherein the heating mechanism comprises a heating pipe fixed inside the container 1, the temperature sensor is used for detecting the temperature inside the container 1 in real time, a temperature signal is transmitted to the controller, and the controller controls the heating pipe to heat the container 1.

The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (4)

1. An oscillation device for producing special enzyme immobilization of 4-aminobutyric acid, which is characterized in that: comprises a container (1), a positioning and supporting mechanism (2) for positioning the container (1) and a driving unit (3) for driving the container (1) to move;

The positioning support mechanism (2) is provided with a strip-shaped offset channel (21) which is parallel to the horizontal plane and has the same width, and a sliding piece (22) which is assembled inside the strip-shaped offset channel (21) and can slide along the length direction of the strip-shaped offset channel (21), wherein a circular through hole (221) for a container (1) to pass through is formed in the middle position of the sliding piece (22);

The container (1) is fitted inside the circular through hole (221) through the circular through hole (221), and the container (1) can axially rotate inside the circular through hole (221);

the driving unit (3) comprises a first driving mechanism (31) for driving the container (1) to reciprocate along the length direction of the strip-shaped offset channel (21) and a second driving mechanism (32) for driving the container (1) to synchronously rotate at a non-uniform speed;

The positioning support mechanism (2) comprises a positioning substrate (23) which is horizontally arranged, the strip-shaped offset channel (21) is formed on the positioning substrate (23), the sliding piece (22) is provided with a abutting edge (222) which is attached to two side walls of the strip-shaped offset channel (21), convex edges (223) are formed by outwards extending the upper end and the lower end of the abutting edge (222) close to the two sides of the strip-shaped offset channel (21), one side of the convex edges (223) opposite to the positioning substrate (23) is uniformly provided with arc-shaped grooves (224), and rolling balls (225) are clamped in the arc-shaped grooves (224);

The container (1) is integrally cylindrical, a mixing cavity is formed in the container (1), a feed inlet (11) is formed in the top of the container (1), a cover body is further arranged at the position of the feed inlet (11), fixing columns (12) are vertically arranged at the axle center positions of the upper end face and the lower end face of the container (1), a U-shaped locating plate (24) is further arranged at the locating support mechanism (2), strip-shaped limiting through holes (241) which are matched with strip-shaped offset channels (21) and are sleeved outside the two fixing columns (12) are formed in the two ends of the U-shaped locating plate (24), the U-shaped locating plate (24) is fixedly connected with a locating base plate (23), a discharge outlet (13) is formed in the bottom of the container (1) in a penetrating mode downwards along the axle center of the container, and a valve is arranged at the discharge outlet (13);

The first driving mechanism (31) comprises lever assemblies (311) symmetrically arranged on the upper end face and the lower end face of the U-shaped positioning plate (24), a connecting assembly (312) used for connecting the lever assemblies (311) and driving the lever assemblies (311) to swing, and a driving source (313); wherein the lever assembly (311) comprises;

the support column (3111) is fixed at one end of the U-shaped positioning plate (24) close to the strip-shaped limiting through hole (241); and

The swing rod (3112) is rotationally connected with the support column (3111), the swing rod (3112) axially rotates along the support column (3111), and a through hole (3113) for the fixed column (12) to pass through is formed in one end, close to the fixed column (12), of the other end of the swing rod (3112);

the connection assembly (312) includes:

The two ends of the fixing rod (3121) are fixedly connected with the other ends of the two swing rods (3112) respectively;

The deflection block (3122) is fixed at the middle part of the fixed rod (3121), and a strip-shaped through groove (3123) is arranged on the deflection block (3122) in a penetrating way; and

The two support plates (3124) are symmetrically arranged on the side walls of the U-shaped positioning plates (24) at the two sides of the deflection block (3122), the opposite sides of the two support plates (3124) are coaxially and rotatably provided with rotating disks (3125), and a force application rod (3126) penetrating through the strip-shaped through groove (3123) is fixed at the edge position between the two rotating disks (3125);

the driving source (313) is used for driving the rotating disc (3125) to rotate circumferentially, the driving source (313) is a speed reducer and a motor, the input end of the speed reducer is fixedly connected with the output end of the motor, and the output end of the speed reducer is fixedly connected with the driving shaft of the rotating disc (3125);

the strip-shaped offset channel (21) is arc-shaped, and the center of the strip-shaped offset channel (21) is positioned on the central axis of the support column (3111);

The second driving mechanism (32) comprises a one-way bearing (321) assembled on the fixed column (12), a first gear (322) assembled outside the one-way bearing (321), and a second gear (323) fixed outside the support column (3111) and meshed with the first gear (322).

2. The oscillation device for producing the immobilized enzyme specific for 4-aminobutyric acid according to claim 1, wherein: the outer side of the positioning substrate (23) is provided with an outer ring barrel (4) which is used for encircling the positioning substrate (23) and the driving unit (3), the outer ring barrel (4) is fixedly connected with the positioning substrate (23), and the bottoms of the outer ring barrels (4) are uniformly provided with supporting legs.

3. The oscillation device for producing the immobilized enzyme specific for 4-aminobutyric acid according to claim 2, wherein: the device is characterized by further comprising a buffer mechanism (5) for slowing down the rotation of the container (1), wherein the buffer mechanism (5) comprises a piston cylinder (51), the piston cylinder (51) is fixed on the inner wall of the outer ring cylinder (4), a piston (52) is slidably arranged in the piston cylinder (51), a piston rod (53) is arranged on one side of the piston (52), a push plate (54) is arranged on one side of the piston (53) extending to the piston cylinder (51), a reset spring (55) is arranged on the other side of the piston (52) away from the piston rod (53), an air inlet pipe (56) and an air outlet pipe (57) are respectively arranged at one end of the piston cylinder (51) away from the piston rod (53), and a first one-way valve (58) and a second one-way valve are respectively arranged on the air outlet pipe (57) and the air inlet pipe;

The buffer mechanism (5) further comprises a baffle (510) arranged on the outer side of the container (1) and a T-shaped air jet pipe (511) coaxially arranged inside the container (1), wherein the top end of the T-shaped air jet pipe (511) penetrates through the top of the container (1) and a fixing column (12) positioned at the top is connected with an air outlet pipe (57) through a hose, and when the container (1) moves, the baffle (510) can push a push plate (54) in a clearance mode to enable a piston (52) to generate displacement.

4. The oscillation device for producing the immobilized enzyme specific for 4-aminobutyric acid according to claim 3, further comprising a heating mechanism comprising a heating pipe fixed inside the container (1), a temperature sensor for detecting the temperature inside the container (1), and a controller.