CN117511725A - Enzyme molecule directed evolution biological information analysis device and method - Google Patents
Enzyme molecule directed evolution biological information analysis device and method Download PDFInfo
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- 238000007622 bioinformatic analysis Methods 0.000 claims 7
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- 230000001276 controlling effect Effects 0.000 claims 1
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- 108010059892 Cellulase Proteins 0.000 description 8
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- 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
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/14—Bioreactors or fermenters specially adapted for specific uses for producing enzymes
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/46—Means for regulation, monitoring, measurement or control, e.g. flow regulation of cellular or enzymatic activity or functionality, e.g. cell viability
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
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Abstract
The invention discloses an enzyme molecule directed evolution biological information analysis device and method, and relates to the technical field of enzyme molecule directed evolution analysis. The invention adopts the enzyme molecular directed evolution sample culture component to realize the environment temperature culture of crude enzyme liquid at a plurality of reaction temperatures at one time; the disposable operation in the temperature interval can be realized, and the orifice plate assembly is conveyed into the sample biological information analysis assembly for analysis and measurement through the sample automatic conveying assembly; the enzyme molecular directed evolution tolerance analysis detection of a plurality of temperature indexes at a large-span temperature can be realized by only performing one-time reaction operation on one-time analysis task, so that the tolerance analysis efficiency of enzyme molecular directed evolution at different temperatures is greatly improved, the analysis uncertainty caused by manual operation is avoided, and the biological interference uncertainty caused by manual repeated operation is reduced.
Description
Technical Field
The invention relates to the technical field of enzyme molecular directed evolution analysis, in particular to an enzyme molecular directed evolution biological information analysis device and method.
Background
Cellulose is the most abundant carbohydrate in the world and is extremely widely distributed and has become the third largest industrial enzyme in the world. With the increasing reduction of global mineral resources, such as: coal, oil, natural gas, etc., cellulose has become an important way to alleviate the global energy crisis as a renewable resource. During the degradation of cellulose by cellulases, glucose is produced, and chemical raw materials and fuels such as: ethanol, acetone, butanol, and the like. The cellulase is utilized to convert a large amount of cellulose resources and cellulose waste into substances required by human life, which has important significance for solving the requirements of people for eating food and living. Currently, the use of cellulases has been spread over various fields such as brewing industry, chemical industry, etc. The application in the food, textile, feed and paper industries is particularly wide.
The cellulase has wide application prospect in various industries, and the application of the cellulase in industry is limited by factors such as enzyme activity, thermal stability, pH stability, cost and the like in recent years. In recent years, with the continuous development of modern molecular biology, in order to improve the enzymatic activity of cellulase, a DNA shuffling technology is utilized to carry out directed evolution research on mutant strains of bacillus amyloliquefaciens (Bacillus amyloliquefaciens strain) subjected to plasma mutagenesis at normal pressure and room temperature, so as to construct a mutant library of escherichia coli. Lays a foundation for further researching the functions and the application of the cellulase at the molecular level.
However, in the process of directed evolution research of cellulase molecules, activity detection analysis is needed to be carried out on the directed evolution cellulase molecules so as to judge and analyze the directed evolution to obtain the tolerance of the enzyme molecules at different temperatures, a 96-well plate is generally used for carrying out reaction before analysis at a set temperature and a pH value, and finally the cultured 96-well plate is manually placed on a bearing plate popped up by an enzyme labeling instrument for analysis and detection.
Therefore, how to automatically and efficiently analyze and detect the enzyme molecular samples of the cellulase analysis and directed evolution is a technical problem which needs to be solved in a matching way.
Disclosure of Invention
In order to solve the technical problems, the invention provides an enzyme molecule directed evolution biological information analysis device and method. The following technical scheme is adopted:
the utility model provides an enzyme molecule directed evolution biological information analysis device, includes enzyme molecule directed evolution sample culture assembly, sample automatic conveying subassembly and sample biological information analysis subassembly, enzyme molecule directed evolution sample culture assembly includes circular bottom plate, orifice plate subassembly and a plurality of independent module of heating, and a plurality of independent modules of heating are evenly installed respectively in the interior bottom of circular bottom plate, orifice plate subassembly includes circular loading board, a plurality of baffle and a plurality of orifice plate, circular loading board cover is put in circular bottom plate, and a plurality of baffles evenly cut apart circular loading board into a plurality of reaction areas, and single reaction area corresponds single independent module of heating, and a plurality of orifice plates are located a plurality of reaction areas respectively, are used for providing different settlement reaction temperature for crude enzyme liquid after the enzyme molecule directed evolution respectively, and sample automatic conveying subassembly is used for taking out the orifice plate subassembly that the reaction is accomplished to input sample biological information analysis subassembly carries out enzyme molecule activity analysis survey.
By adopting the technical scheme, for the analysis and detection of the tolerance of the enzyme molecules under different temperatures of the directed evolution, the most intuitive way is to add the crude enzyme solution after the directed evolution into low-grade solutions under different temperatures for reaction, for example, the tolerance of the enzyme molecules under the temperature range of 40 ℃ to 90 ℃ obtained after the directed evolution of the enzyme molecules needs to be inspected, sample reaction at the temperature range of 40 ℃ is needed to be carried out every 10 ℃, for example, the pore plate is placed in an enzyme labeling instrument for optical calibration after the reaction is finished, so as to feed back the activity degree of the enzyme molecules after the reaction under the temperature of 40 ℃, thus reflecting the tolerance of the enzyme molecules under the temperature of 40 ℃ after the directed evolution, and the analysis and detection can be completed once just like 5 times.
Different from the mode that a common enzyme-labeled instrument needs multiple reactions and multiple determinations, the environment temperature culture of crude enzyme liquid at multiple reaction temperatures is realized at one time by adopting an enzyme molecule directed evolution sample culture component;
the crude enzyme liquid is equally divided into a plurality of parts and is respectively injected into a plurality of reaction areas of the pore plate component, the plurality of reaction areas are respectively and oppositely heated through a plurality of independent heating modules, disposable operation in a temperature interval can be realized, the crude enzyme liquid is automatically reacted at a set temperature, after the reaction is completed in a set time, the pore plate component is conveyed into a sample biological information analysis component together through a sample automatic conveying component for analysis and measurement, and the sample biological information analysis component can be a biological information optical analysis instrument such as an enzyme-labeling instrument.
The enzyme molecular directed evolution tolerance analysis detection of a plurality of temperature indexes at a large-span temperature can be realized by only performing one-time reaction operation on one-time analysis task, so that the tolerance analysis efficiency of enzyme molecular directed evolution at different temperatures is greatly improved, the analysis uncertainty caused by manual operation is avoided, and the biological interference uncertainty caused by manual repeated operation is reduced.
Optionally, the sample biological information analysis assembly comprises a detection shell, a sample injection assembly, a rotary enzyme-labeling instrument measurement module and a measurement data output module, wherein the sample injection assembly comprises an electric pop-up mechanism and a detection end circular supporting plate, an opening is formed in one side of the detection shell, a detection cavity is formed in the detection shell, the electric pop-up mechanism is arranged at the opening of the detection shell, the detection end circular supporting plate is arranged at a pop-up part of the electric pop-up mechanism and is popped up or retracted along with the pop-up part, and the detection end circular supporting plate is used for placing a circular bearing plate;
the rotary enzyme-linked immunosorbent assay instrument comprises an electric rotary table, a C-shaped plate, an assay light source module, a photoelectric signal detector and a photoelectric signal processor, wherein a base of the electric rotary table is arranged on the inner wall of an assay shell, the top of the C-shaped plate is detachably arranged on a rotating part of the electric rotary table, and when the detection end circular supporting plate is retracted, the circular supporting plate is positioned in the middle of the C-shaped plate, the assay light source module is arranged on the inner side of one side of the C-shaped plate, the photoelectric signal detector is arranged on the inner side of the other side of the C-shaped plate and is used for receiving light rays transmitted by the assay light source module through the circular supporting plate, the photoelectric signal detector is in communication connection with the photoelectric signal processor, and the data output end of the photoelectric signal processor is in communication connection with the assay data output module.
By adopting the technical scheme, the sample biological information analysis assembly adopts the sample injection assembly to realize automatic sample injection operation, the sample automatic conveying assembly places the pore plate assembly with the reacted completion on the detection end circular supporting plate of the electric pop-up mechanism in the open state, the electric pop-up mechanism can retract to realize automatic sample injection, and the electric pop-up mechanism is a structure similar to an electric drawer;
after sample injection, the measuring module of the rotary enzyme-labeled instrument starts to act, the electric turntable drives the C-shaped plate to rotate, each time the electric turntable reaches the center of one reaction area, the measuring light source module emits measuring monochromatic light, the photoelectric signal detector receives the light transmitted by the measuring light source module through the pore plate assembly, and the received data signals are transmitted outwards through the measuring data output module, so that optical analysis is carried out on a plurality of samples respectively through one sample injection.
Optionally, the circular bearing plate, the plurality of partition plates and the plurality of pore plates of the pore plate assembly are molded by adopting an optical transparent pure polystyrene material integrated mold.
By adopting the technical scheme, the optical transparent pure polystyrene material can be used for conveniently measuring the transmission of monochromatic light rays emitted by the light source module, and the enzyme molecular reaction can not be interfered after the sterilization treatment by means such as gamma rays.
Optionally, the orifice plate assembly further comprises a plurality of heat-insulating and light-insulating patches, the plurality of heat-insulating and light-insulating patches are respectively stuck to two side surfaces of the plurality of clapboards, and the heat-insulating and light-insulating patches are made of black polypropylene materials.
By adopting the technical scheme, the heat-insulating light-insulating paste made of the black polypropylene material can better enable the round bearing plate to be uniformly divided into a plurality of reaction areas by the plurality of partition plates, the temperatures in the reaction areas are not affected mutually, and the reflection and refraction effects are basically avoided during optical measurement.
Optionally, the independent heating module includes heating wire, soaking plate, temperature sensor and temperature control chip, the heating wire is installed in the heater strip installation cavity that circular bottom plate top surface was preset, the soaking plate is installed at circular bottom plate top surface, is located the heating wire top, and the soaking plate is single reaction zone and heats alone, temperature sensor detects the temperature value in the single reaction zone to transmit to the temperature control chip, the operating power of temperature control chip according to temperature sensor survey temperature and settlement temperature control heating wire.
Through adopting above-mentioned technical scheme, the independent module of heating is based on the heating wire and heats, evenly transmits the orifice plate of reaction region top with the temperature of heating wire through the vapor chamber, and the temperature control chip can realize carrying out constant temperature control to the reaction liquid in the orifice plate on the reaction region to the control of heating wire operating power through temperature sensor's detection feedback.
Optionally, the electronic pop-up mechanism is electronic linear slide rail, and the middle part of the circular layer board of detection end sets up circular fretwork region, and when the orifice plate subassembly was located the circular layer board of detection end, circular fretwork region covered a plurality of reaction areas, and when the C template of electronic carousel drive rotated, the monochromatic light that survey light source module sent was passed through circular loading board to it is received by photoelectric signal detector to see through circular fretwork region.
Through adopting above-mentioned technical scheme, the middle part of the circular layer board of detection end sets up circular fretwork area, can be convenient for survey the monochromatic light that light source module sent and see through circular loading board and receive by photoelectric signal detector.
Optionally, the automatic transport subassembly of sample includes sideslip electronic slide rail, flexible electric cylinder and electronic clamping jaw, sideslip electronic slide rail includes track and electronic slider, the track erects in the directional evolution sample of enzyme molecule cultivates subassembly and sample biological information analysis subassembly top, electronic slider is located the track and removes, the cylinder body of flexible electric cylinder is installed on electronic slider, follows electronic slider lateral shifting, the casing of electronic clamping jaw is installed on the piston rod of flexible electric cylinder, and the top of circular loading board is equipped with grabs the board, and electronic clamping jaw grabs the clamp and grabs orifice plate subassembly in grabbing the board department.
Through adopting above-mentioned technical scheme, the cooperation action of sideslip electronic slide rail, flexible electronic jar and electronic clamping jaw can realize the automation of circular loading board and snatch and sideslip.
Optionally, an electric cover plate is arranged at the top of the round bottom supporting plate.
By adopting the technical scheme, the electric cover plate can be automatically opened and closed, so that the pore plate assembly reacts in a closed environment.
Optionally, the sample biological information analysis component further comprises a computer, wherein the computer is in communication connection with the measurement data output module, and the computer is preloaded with ELISA Reader analysis software.
By adopting the technical scheme, the ELISA Reader is enzyme-labeled instrument software with strong function and simple operation. The ELISA Reader can be operated by a computer to accurately and rapidly read and analyze the experimental data of the ELISA Reader, and generate a related report and chart, so that an experimenter can rapidly judge the experimental result.
The computer can also respectively control the execution actions of each electric actuator of the enzyme molecule directed evolution sample culture assembly, the sample automatic conveying assembly and the sample biological information analysis assembly, so as to realize automatic control.
The enzyme molecular directed evolution biological information analysis method adopts an enzyme molecular directed evolution biological information analysis device to carry out enzyme molecular directed evolution analysis, and the specific method comprises the following steps:
step 1, opening an electric cover plate, respectively adding equal amounts of crude enzyme solution and substrate solution into a plurality of reaction areas of an orifice plate assembly, and respectively adjusting the reaction temperatures in the plurality of reaction areas through a plurality of independent heating modules according to reaction requirements;
step 2, covering an electric cover plate, and reacting for a set time;
step 3, an electric sliding block is started to drive a telescopic electric cylinder and an electric clamping jaw to be right above the pore plate component, an electric cover plate is opened, a piston rod of the telescopic electric cylinder extends out, the electric clamping jaw clamps the pore plate component at a grabbing plate, the piston rod of the telescopic electric cylinder retracts, the electric sliding block moves again, and the pore plate component moves to be above the sample biological information analysis component;
step 4, the electric pop-up mechanism pops up, the telescopic electric cylinder and the electric clamping jaw are linked to place the pore plate assembly on the circular supporting plate at the detection end, and the electric pop-up mechanism retracts;
and 5, starting a measuring module of the rotary enzyme-linked immunosorbent assay instrument, driving the C-shaped plate to rotate by an electric turntable, transmitting measuring monochromatic light to the center of each reaction area by the measuring light source module, receiving the light transmitted by the measuring light source module through the pore plate assembly by the photoelectric signal detector, transmitting the received data signal to a computer through a measuring data output module, and analyzing the measuring data by the computer through ELISA Reader analysis software and outputting an analysis result.
In summary, the present invention includes at least one of the following beneficial technical effects:
the invention can provide a device and a method for analyzing biological information of directed evolution of enzyme molecules, which are different from the mode that a common enzyme-labeled instrument needs multiple reactions and multiple determinations, and adopt an enzyme molecule directed evolution sample culture component to realize the environment temperature culture of crude enzyme liquid at multiple reaction temperatures at one time; the disposable operation in the temperature interval can be realized, the reaction is automatically carried out at the set temperature, and after the reaction is finished for the set time, the pore plate component is conveyed into the sample biological information analysis component for analysis and measurement through the sample automatic conveying component; the enzyme molecular directed evolution tolerance analysis detection of a plurality of temperature indexes at a large-span temperature can be realized by only performing one-time reaction operation on one-time analysis task, so that the tolerance analysis efficiency of enzyme molecular directed evolution at different temperatures is greatly improved, the analysis uncertainty caused by manual operation is avoided, and the biological interference uncertainty caused by manual repeated operation is reduced.
Drawings
FIG. 1 is a schematic diagram of the structure of an enzyme molecular directed evolution sample culture assembly of an enzyme molecular directed evolution biological information analysis device of the present invention;
FIG. 2 is a schematic diagram showing a top view structure of an enzyme molecular directed evolution sample culture assembly of the enzyme molecular directed evolution biological information analysis device of the present invention;
FIG. 3 is a schematic diagram showing the structural arrangement of an enzyme molecule directed evolution biological information analysis device;
FIG. 4 is a schematic diagram showing the internal structure of a sample bioinformation analysis module of an enzyme molecule directed evolution bioinformation analysis device according to the present invention;
FIG. 5 is a schematic diagram of a measurement module of a rotary microplate reader of a sample bioinformation analysis component of an enzyme molecule directed evolution bioinformation analysis device according to the present invention;
FIG. 6 is a schematic diagram of the connection principle of the electric device of the enzyme molecule directed evolution biological information analysis device.
Reference numerals illustrate: 1. an enzyme molecule directed evolution sample culture component; 11. a circular bottom plate; 111. an electric cover plate; 121. a circular bearing plate; 122. a partition plate; 123. an orifice plate; 131. heating wires; 132. a soaking plate; 133. a temperature sensor; 134. a temperature control chip; 2. a sample bioinformation analysis component; 21. a sample injection assembly; 211. an electric pop-up mechanism; 212. a circular supporting plate at the detection end; 22. a rotary enzyme-labeling instrument measuring module; 221. an electric turntable; 222. a C-shaped plate; 223. measuring a light source module; 224. a photoelectric signal detector; 225. an optoelectronic signal processor; 23. a measurement data output module; 24. a detection housing; 25. a computer; 311. a track; 312. an electric slide block; 32. a telescopic electric cylinder; 33. and (5) an electric clamping jaw.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings.
The embodiment of the invention discloses an enzyme molecule directed evolution biological information analysis device and method.
Referring to fig. 1-6, embodiment 1, an enzyme molecule directed evolution biological information analysis device, which comprises an enzyme molecule directed evolution sample culturing component 1, a sample automatic conveying component and a sample biological information analysis component 2, wherein the enzyme molecule directed evolution sample culturing component 1 comprises a circular bottom support plate 11, a pore plate component and a plurality of independent heating modules, the independent heating modules are respectively and uniformly arranged at the inner bottom of the circular bottom support plate 11, the pore plate component comprises a circular bearing plate 121, a plurality of partition plates 122 and a plurality of pore plates 123, the circular bearing plate 121 is sleeved in the circular bottom support plate 11, the circular bearing plate 121 is uniformly divided into a plurality of reaction areas by the plurality of partition plates 122, the single reaction area corresponds to the single independent heating module, the plurality of pore plates 123 are respectively positioned in the plurality of reaction areas and are respectively used for providing different set reaction temperatures for crude enzyme liquid after enzyme molecule directed evolution, and the sample automatic conveying component is used for taking out the pore plate component after reaction completion and inputting the pore plate component into the sample biological information analysis component 2 for enzyme molecule activity analysis and measurement.
For the analysis and detection of the tolerance of the enzyme molecules under different temperatures of the directed evolution, the most intuitive way is to add the crude enzyme solution after the directed evolution into low-grade solutions under different temperatures for reaction, for example, the tolerance of the enzyme molecules obtained after the directed evolution of the enzyme molecules in the range of 40 ℃ to 90 ℃ needs to be examined, sample reactions at 40 ℃ are required to be carried out at intervals of set temperature, for example, at intervals of 10 ℃, after the reaction is finished, the pore plate is placed in an enzyme labeling instrument for optical calibration so as to feed back the activity degree of the enzyme molecules after the reaction at 40 ℃, thus reflecting the tolerance of the enzyme molecules to the temperature of 40 ℃ after the directed evolution, and the analysis and detection at once can be completed only by 5 times.
Different from the mode that a common enzyme-labeled instrument needs multiple reactions and multiple determinations, the enzyme molecular directed evolution sample culture assembly 1 is adopted to realize the environment temperature culture of crude enzyme liquid at multiple reaction temperatures at one time;
the crude enzyme liquid is equally divided into a plurality of parts and is respectively injected into a plurality of reaction areas of the pore plate component, the plurality of reaction areas are respectively and oppositely heated through a plurality of independent heating modules, disposable operation in a temperature interval can be realized, the reaction is automatically carried out at a set temperature, after the reaction is completed in a set time, the pore plate component is conveyed into the sample biological information analysis component 2 together through the sample automatic conveying component for analysis and measurement, and the sample biological information analysis component 2 can be a biological information optical analysis instrument such as an enzyme-labeling instrument.
The enzyme molecular directed evolution tolerance analysis detection of a plurality of temperature indexes at a large-span temperature can be realized by only performing one-time reaction operation on one-time analysis task, so that the tolerance analysis efficiency of enzyme molecular directed evolution at different temperatures is greatly improved, the analysis uncertainty caused by manual operation is avoided, and the biological interference uncertainty caused by manual repeated operation is reduced.
In embodiment 2, the sample biological information analysis assembly 2 comprises a detection housing 24, a sample feeding assembly 21, a rotary enzyme-labeled instrument measurement module 22 and a measurement data output module 23, wherein the sample feeding assembly 21 comprises an electric pop-up mechanism 211 and a detection end circular support plate 212, an opening is formed in one side of the detection housing 24, a detection cavity is formed inside the detection housing, the electric pop-up mechanism 211 is arranged at the opening of the detection housing 24, the detection end circular support plate 212 is arranged at the pop-up part of the electric pop-up mechanism 211 and pops up or withdraws back along with the pop-up part, and the detection end circular support plate 212 is used for placing a circular bearing plate 121;
the rotary enzyme-labeled instrument measuring module 22 comprises an electric rotary table 221, a C-shaped plate 222, a measuring light source module 223, a photoelectric signal detector 224 and a photoelectric signal processor 225, wherein a base of the electric rotary table 221 is arranged on the inner wall of the detecting shell 24, the top of the C-shaped plate 222 is detachably arranged on a rotating part of the electric rotary table 221, and when the detecting end circular supporting plate 212 is retracted, the circular supporting plate 121 is positioned in the middle of the C-shaped plate 222, the measuring light source module 223 is arranged on the inner side of one side of the C-shaped plate 222, the photoelectric signal detector 224 is arranged on the inner side of the other side of the C-shaped plate 222 and is used for receiving light rays transmitted by the measuring light source module 223 through the circular supporting plate 121, the photoelectric signal detector 224 is in communication connection with the photoelectric signal processor 225, and a data output end of the photoelectric signal processor 225 is in communication connection with the measuring data output module 23.
The sample biological information analysis assembly 2 adopts a sample injection assembly 21 to realize automatic sample injection operation, the sample automatic conveying assembly places the pore plate assembly with the reacted completion on a circular supporting plate 212 at the detection end of an electric pop-up mechanism 211 in an opened state, the electric pop-up mechanism 211 is retracted to realize automatic sample injection, and the electric pop-up mechanism 211 is a structure similar to an electric drawer;
after sample injection, the rotary enzyme-labeled instrument measuring module 22 is started, the electric rotary table 221 drives the C-shaped plate 222 to rotate, the measuring light source module 223 emits measuring monochromatic light every time the center of one reaction area is reached, the photoelectric signal detector 224 receives the light transmitted by the measuring light source module 223 through the pore plate assembly, and the received data signals are transmitted outwards through the measuring data output module 23, so that optical analysis is carried out on a plurality of samples respectively through one sample injection.
Embodiment 3 the circular carrier plate 121, the plurality of spacers 122 and the plurality of well plates 123 of the well plate assembly are molded using an optically transparent pure polystyrene material integral mold.
The optically transparent pure polystyrene material can be used for conveniently measuring the transmission of monochromatic light rays emitted by the light source module 223, and the enzyme molecular reaction can not be interfered after sterilization treatment by means of gamma rays and the like.
Embodiment 4, the orifice plate assembly further comprises a plurality of heat-insulating and light-insulating patches, wherein the plurality of heat-insulating and light-insulating patches are respectively adhered to two side surfaces of the plurality of partition plates 122, and the heat-insulating and light-insulating patches are made of black polypropylene.
The black polypropylene thermal insulation paste can better enable the plurality of partition plates 122 to uniformly divide the round bearing plate 121 into a plurality of reaction areas, and the temperatures in the reaction areas do not influence each other and basically have no reflection and refraction effects in optical measurement.
Embodiment 5, the independent heating module includes heating wire 131, soaking plate 132, temperature sensor 133 and temperature control chip 134, and heating wire 131 installs in the heater strip installation cavity that circular bottom plate 11 top surface was preset, and soaking plate 132 installs at circular bottom plate 11 top surface, is located heating wire 131 top, and soaking plate 132 is single reaction zone alone and heats, and temperature sensor 133 detects the temperature value in the single reaction zone to transmit to temperature control chip 134, temperature control chip 134 is according to temperature sensor 133 survey temperature and the operating power of settlement temperature control heating wire 131.
The independent heating module is based on heating wire 131 to heat, evenly transmits the temperature of heating wire 131 to orifice plate 123 above the reaction area through vapor chamber 132, and then through the detection feedback of temperature sensor 133, temperature control chip 134 is to the control of heating wire 131 working power, can realize the constant temperature control to the reaction liquid in the orifice plate 123 on the reaction area.
In embodiment 6, the electric pop-up mechanism 211 is an electric linear slide rail, a circular hollow area is disposed in the middle of the circular support plate 212 at the detection end, when the orifice plate assembly is located on the circular support plate 212 at the detection end, the circular hollow area covers a plurality of reaction areas, and when the electric turntable 221 drives the C-shaped plate 222 to rotate, the monochromatic light emitted by the measurement light source module 223 passes through the circular bearing plate 121 and is received by the photoelectric signal detector 224 through the circular hollow area.
The middle part of the detection end circular supporting plate 212 is provided with a circular hollowed-out area, so that the monochromatic light emitted by the measurement light source module 223 can be conveniently received by the photoelectric signal detector 224 through the circular bearing plate 121.
Embodiment 7, the automatic sample conveying assembly includes sideslip electronic slide rail, flexible electronic jar 32 and electronic clamping jaw 33, sideslip electronic slide rail includes track 311 and electronic slider 312, track 311 erects in the directional evolution sample of enzyme molecule cultivates subassembly 1 and sample biological information analysis subassembly 2 top, electronic slider 312 is located the track 311 and removes, the cylinder body of flexible electronic jar 32 is installed on electronic slider 312, follow electronic slider 312 lateral shifting, the casing of electronic clamping jaw 33 is installed on the piston rod of flexible electronic jar 32, the top of circular loading board 121 is equipped with the grabbing plate, electronic clamping jaw 33 grabs the clamp and grabs the orifice plate subassembly in grabbing the plate department.
The automatic grabbing and traversing of the circular bearing plate 121 can be realized by the matched actions of the traversing electric slide rail, the telescopic electric cylinder 32 and the electric clamping jaw 33.
In embodiment 8, an electric cover plate 111 is provided on top of the circular bottom plate 11.
The motorized cover 111 may be automatically opened and closed to allow the orifice plate assembly to react in a closed environment.
In example 9, the sample bioinformation analysis module 2 further comprises a computer 25, the computer 25 is communicatively connected to the assay data output module 23, and the computer 25 is preloaded with ELISA Reader analysis software.
ELISA Reader is a powerful, easy-to-operate enzyme-labeled instrument software. The computer 25 runs the ELISA Reader to accurately and rapidly read and analyze the experimental data of the ELISA Reader, and generate related reports and charts, so that an experimenter can rapidly judge the experimental result.
The computer 25 can also respectively control the execution actions of the electric actuators of the enzyme molecule directed evolution sample culture assembly 1, the sample automatic conveying assembly and the sample biological information analysis assembly 2, so as to realize automatic control.
Embodiment 10, an enzyme molecule directed evolution biological information analysis method, which uses an enzyme molecule directed evolution biological information analysis device to perform enzyme molecule directed evolution analysis, specifically comprises the following steps:
step 1, opening an electric cover plate 111, respectively adding equal amounts of crude enzyme solution and substrate solution into a plurality of reaction areas of an orifice plate assembly, and respectively adjusting the reaction temperatures in the plurality of reaction areas through a plurality of independent heating modules according to reaction requirements;
step 2, covering an electric cover plate 111, and reacting for a set time;
step 3, the electric sliding block 312 starts to drive the telescopic electric cylinder 32 and the electric clamping jaw 33 to be right above the pore plate component, the electric cover plate 111 is opened, a piston rod of the telescopic electric cylinder 32 extends out, the electric clamping jaw 33 clamps the pore plate component at the grabbing plate, the piston rod of the telescopic electric cylinder 32 retracts, the electric sliding block 312 moves again, and the pore plate component is moved to be above the sample biological information analysis component 2;
step 4, the electric popup mechanism 211 popups, the telescopic electric cylinder 32 and the electric clamping jaw 33 are linked to place the orifice plate assembly on the circular supporting plate 212 at the detection end, and the electric popup mechanism 211 retracts;
step 5, the rotary microplate Reader measuring module 22 is started, the electric turntable 221 drives the C-shaped plate 222 to rotate, each time the center of one reaction area is reached, the measuring light source module 223 emits measuring monochromatic light, the photoelectric signal detector 224 receives the light transmitted by the measuring light source module 223 through the orifice plate assembly, the received data signal is transmitted to the computer 25 through the measuring data output module 23, and the computer 25 operates ELISA Reader analyzing software to analyze the measuring data and output an analyzing result.
The above embodiments are not intended to limit the scope of the present invention, and therefore: all equivalent changes in structure, shape and principle of the invention should be covered in the scope of protection of the invention.
Claims (10)
1. An enzyme molecule directed evolution biological information analysis device is characterized in that: the enzyme molecular directional evolution sample culture device comprises an enzyme molecular directional evolution sample culture assembly (1), a sample automatic conveying assembly and a sample biological information analysis assembly (2), wherein the enzyme molecular directional evolution sample culture assembly (1) comprises a circular bottom support plate (11), an orifice plate assembly and a plurality of independent heating modules, the independent heating modules are respectively and uniformly arranged at the inner bottoms of the circular bottom support plate (11), the orifice plate assembly comprises a circular bearing plate (121), a plurality of partition plates (122) and a plurality of orifice plates (123), the circular bearing plate (121) is sleeved in the circular bottom support plate (11), the circular bearing plate (121) is uniformly divided into a plurality of reaction areas by the plurality of partition plates (122), the single reaction area corresponds to the single independent heating module, the orifice plates (123) are respectively positioned in the plurality of reaction areas and are respectively used for providing different set reaction temperatures for crude enzyme liquid after the directional evolution of enzyme molecules, and the sample automatic conveying assembly is used for taking out the orifice plate assembly after the reaction is completed and inputting the sample biological information analysis assembly (2) for enzyme molecular activity analysis and measurement.
2. The enzyme molecule directed evolution bioinformatic analysis apparatus of claim 1 wherein: the sample biological information analysis assembly (2) comprises a detection shell (24), a sample injection assembly (21), a rotary enzyme-labeling instrument measurement module (22) and a measurement data output module (23), wherein the sample injection assembly (21) comprises an electric ejection mechanism (211) and a detection end circular supporting plate (212), an opening is formed in one side of the detection shell (24), a detection cavity is formed in the detection shell, the electric ejection mechanism (211) is arranged at the opening of the detection shell (24), the detection end circular supporting plate (212) is arranged at the ejection part of the electric ejection mechanism (211) and is ejected or retracted along with the ejection part, and the detection end circular supporting plate (212) is used for placing a circular bearing plate (121);
the utility model provides a rotatory ELIAS appearance survey module (22) includes electric turntable (221), C template (222), survey light source module (223), photoelectric signal detector (224) and photoelectric signal processor (225), the pedestal mounting of electric turntable (221) is on detecting casing (24) inner wall, the top demountable installation of C template (222) is on the rotation part of electric turntable (221), and when detecting end circular layer board (212) is withdrawn, circular loading board (121) is located the middle part of C template (222), survey light source module (223) are installed in the inboard of one side of C template (222), photoelectric signal detector (224) are installed in the inboard of the another side of C template (222) for receive the light of survey light source module (223) through circular loading board (121), photoelectric signal detector (224) and photoelectric signal processor (225) communication connection, the data output end and the survey data output module (23) of photoelectric signal processor (225).
3. The enzyme molecule directed evolution bioinformatic analysis apparatus of claim 1 wherein: the circular bearing plate (121), the plurality of partition plates (122) and the plurality of pore plates (123) of the pore plate assembly are molded by adopting an optical transparent pure polystyrene material integrated mold.
4. The enzyme molecule directed evolution biological information analysis apparatus according to claim 3, wherein: the pore plate assembly further comprises a plurality of heat-insulating and light-insulating pastes, the heat-insulating and light-insulating pastes are respectively adhered to two side surfaces of the plurality of partition plates (122), and the heat-insulating and light-insulating pastes are made of black polypropylene materials.
5. The enzyme molecule directed evolution bioinformatic analysis apparatus of claim 1 wherein: the independent heating module comprises an electric heating wire (131), a soaking plate (132), a temperature sensor (133) and a temperature control chip (134), wherein the electric heating wire (131) is installed in a heating wire installation cavity preset in the top surface of the round bottom supporting plate (11), the soaking plate (132) is installed on the top surface of the round bottom supporting plate (11) and is located above the electric heating wire (131), the soaking plate (132) is independently used for heating a single reaction area, the temperature sensor (133) detects a temperature value in the single reaction area and transmits the temperature value to the temperature control chip (134), and the temperature control chip (134) is used for controlling the working power of the electric heating wire (131) according to the measured temperature and the set temperature of the temperature sensor (133).
6. The enzyme molecule directed evolution bioinformatic analysis apparatus of claim 2 wherein: the electric pop-up mechanism (211) is an electric linear slide rail, a circular hollow area is arranged in the middle of the circular supporting plate (212) at the detection end, when the pore plate component is positioned on the circular supporting plate (212) at the detection end, the circular hollow area covers a plurality of reaction areas, and when the electric turntable (221) drives the C-shaped plate (222) to rotate, monochromatic light emitted by the measuring light source module (223) penetrates through the circular bearing plate (121) and is received by the photoelectric signal detector (224) through the circular hollow area.
7. The enzyme molecule directed evolution bioinformatic analysis apparatus of claim 2 wherein: the automatic sample conveying assembly comprises a transverse moving electric sliding rail, a telescopic electric cylinder (32) and an electric clamping jaw (33), wherein the transverse moving electric sliding rail comprises a rail (311) and an electric sliding block (312), the rail (311) is arranged above an enzyme molecule directional evolution sample culturing assembly (1) and a sample biological information analyzing assembly (2), the electric sliding block (312) is located on the rail (311) and moves, a cylinder body of the telescopic electric cylinder (32) is mounted on the electric sliding block (312) and moves transversely along with the electric sliding block (312), a shell of the electric clamping jaw (33) is mounted on a piston rod of the telescopic electric cylinder (32), a grabbing plate is arranged at the top of a circular bearing plate (121), and the electric clamping jaw (33) grabs a pore plate assembly at the grabbing plate.
8. The enzyme molecule directed evolution bioinformatic analysis apparatus of claim 7 wherein: an electric cover plate (111) is arranged at the top of the round bottom supporting plate (11).
9. The enzyme molecule directed evolution bioinformatic analysis apparatus of claim 8 wherein: the sample biological information analysis component (2) further comprises a computer (25), the computer (25) is in communication connection with the measurement data output module (23), and the computer (25) is preloaded with ELISA Reader analysis software.
10. An enzyme molecule directed evolution biological information analysis method is characterized in that: the enzyme molecular directed evolution biological information analysis device of claim 9 is adopted for enzyme molecular directed evolution analysis, and the specific method is as follows:
step 1, an electric cover plate (111) is opened, equal parts of crude enzyme solution and substrate solution are respectively added into a plurality of reaction areas of an orifice plate assembly, and the reaction temperatures in the plurality of reaction areas are respectively regulated by a plurality of independent heating modules according to the reaction requirements;
step 2, covering an electric cover plate (111) to react for a set time;
step 3, an electric sliding block (312) starts to drive a telescopic electric cylinder (32) and an electric clamping jaw (33) to be right above the pore plate component, an electric cover plate (111) is opened, a piston rod of the telescopic electric cylinder (32) stretches out, the electric clamping jaw (33) clamps the pore plate component at a grabbing plate, the piston rod of the telescopic electric cylinder (32) retracts, the electric sliding block (312) moves again, and the pore plate component is moved to be above the sample biological information analysis component (2);
step 4, the electric pop-up mechanism (211) pops up, the telescopic electric cylinder (32) and the electric clamping jaw (33) are linked to place the pore plate assembly on the circular supporting plate (212) at the detection end, and the electric pop-up mechanism (211) retracts;
and 5, starting a rotary enzyme-labeled instrument measuring module (22), driving a C-shaped plate (222) to rotate by an electric rotary table (221), emitting measuring monochromatic light by a measuring light source module (223) every time the center of a reaction area is reached, receiving light rays transmitted by the measuring light source module (223) through an orifice plate assembly by a photoelectric signal detector (224), transmitting received data signals to a computer (25) through a measuring data output module (23), and analyzing measuring data by the computer (25) by operating ELISA Reader analysis software, and outputting an analysis result.
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