CN110938518A - Filtering device and method for microbial enzyme activity test - Google Patents
Filtering device and method for microbial enzyme activity test Download PDFInfo
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- CN110938518A CN110938518A CN201911018518.7A CN201911018518A CN110938518A CN 110938518 A CN110938518 A CN 110938518A CN 201911018518 A CN201911018518 A CN 201911018518A CN 110938518 A CN110938518 A CN 110938518A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/24—Methods of sampling, or inoculating or spreading a sample; Methods of physically isolating an intact microorganisms
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Abstract
The invention provides a filtering device and a method for microbial enzyme activity test, the device comprises a liquid supply mechanism for providing liquid to be filtered and a collecting mechanism for collecting filtrate, the liquid supply mechanism comprises an upper connecting table, the collecting mechanism comprises a collecting container, a lower connecting table for placing a filter membrane is arranged on the collecting container, the upper connecting table and the lower connecting table are detachably connected, at least one filtering channel is arranged on the upper connecting table and the lower connecting table in a penetrating manner, the liquid to be filtered in the liquid supply mechanism passes through the filtering channel and the filter membrane and enters a liquid collecting cavity of the collecting mechanism, and trapped matters are enriched on the filter membrane. The device of the invention enriches cells by a filtering mode, the cell sap consumption required to be filtered is obviously reduced, the number of single filtering samples is obviously increased, and further, the substrates required in the subsequent enzyme activity test are obviously reduced.
Description
Technical Field
The invention relates to the technical field of microbial enzyme activity test, in particular to a filtering device and a filtering method for microbial enzyme activity test.
Background
The halogenated organic pollutants have the characteristics of strong durability and difficult biodegradation, and mainly comprise perfluoro or Partial Fluoro Compounds (PFCs), Chlorinated Organic Compounds (COCs) and Brominated Organic Compounds (BOCs). Although organic halides are often used as raw materials, intermediates, solvents and the like in organic synthesis, the organic halides have significant effects in human production and life. However, many organic halides are discharged into the environment, either randomly or inevitably, and pose serious hazards to the ozone layer, ecological safety and human health.
The use and discharge of a large amount of halogenated organic matters cause the pollution of the halogenated matters in the water body to be increasingly serious, and threaten the ecological safety and the human health. Moreover, the halogenated organic pollutants have the characteristics of environmental persistence, difficult biodegradation, bioaccumulation, high toxicity, long-distance migration capability and the like, and are distributed in the field environments such as soil, atmosphere and the like, so that how to effectively solve the problem of halogenated pollutants becomes the focus of attention in the field of environment. At present, the methods for repairing the soil and underground water polluted by the halogenated organic pollutants mainly comprise physical repair, chemical repair and microbial repair. The anaerobic microorganism dehalogenation process is one of the most potential in-situ remediation methods for the pollution of the environmental halogenated organic matters at present due to the advantages of no secondary pollution, low cost, relatively high environmental friendliness compared with physical and chemical dehalogenation methods and the like. The process of dehalogenation using anaerobic microorganisms is essentially an enzymatic reaction catalyzed by dehalogenases, and thus a large number of experiments are required to test the dehalogenase activity.
The microorganism in-vitro enzyme activity experiment is one of means for researching the catalytic reaction of the microorganism enzyme, can test the catalytic activity of the microorganism enzyme to a substrate in a short time and research the functional characteristics of specific enzyme for catalyzing the conversion of the substrate; by adjusting the reaction conditions, the mechanism of utilization of the microbial substrate can be studied. Taking the existing in vitro enzyme activity test of microbial cells as an example, as shown in fig. 1, 1 liter of cell bacterial liquid is subpackaged into centrifuge tubes for centrifugation (10000 × g, 5 minutes, 4 ℃), supernatant is removed, 3mL of supernatant is left in each centrifuge tube, precipitates are uniformly mixed, secondary centrifugation (15000 × g, 5 minutes, 4 ℃) is carried out, supernatant is removed, circulation is carried out (generally, centrifugation is carried out for twenty times, the time is about 1-2 hours or more) until 1 liter of cell bacterial liquid is completely used up, finally, cell bacterial liquids of the centrifuge tubes are combined, the cell bacterial liquid with the volume of about 0.1mL and the enzyme concentration of 100-. The method has the advantages of complex operation, limited cell centrifugally collected, long time consumption, low flux, large test reaction system and large consumption of consumables, so an enzyme activity test technology which is convenient to operate, low in manufacturing cost and high in cell recovery rate needs to be developed so as to further research the microbial enzymatic reaction.
The existing vacuum filtration device has unreasonable structural design and cannot be used for cell separation in microorganism in-vitro enzyme activity experiments.
Disclosure of Invention
The invention aims to overcome the defects and shortcomings of the prior art and provide a filtering device and a method for microbial enzyme activity test, and the filtering device and the method overcome the defects of complex operation of enzyme activity test, low recovery rate of centrifugally collected cells, long time consumption, low flux, large test reaction system, large consumption of consumables and the like in the existing microbial dehalogenation experiment, particularly aim at dehalogenation bacteria with extremely low microbial growth amount, react each glass fiber membrane enriched with cells with a trace substrate (0.4mL) by using a suction membrane filtering mode, and can realize that extremely small amount (each tube is less than or equal to 10mL) of cell sap simultaneously and quickly enrich different microbial extracellular enzymes so as to test the enzyme activity under different substrates.
The above purpose of the invention is realized by the following technical scheme:
the utility model provides a filter equipment for microbial enzyme test alive, is including being used for providing the confession liquid mechanism of waiting to filter liquid, the collection mechanism that is used for collecting filtrating, supply liquid mechanism to include and connect the platform, collect the mechanism including collecting the container, be equipped with the lower platform that connects that is used for placing the filter membrane on the collection container, go up and connect the platform, connect the platform down and can dismantle the connection, go up and connect the platform, connect the bench down and run through and be equipped with at least one filtration passageway, supply the liquid that waits to filter in the liquid mechanism and pass filtration passageway and the filter membrane gets into collect the liquid collecting cavity of mechanism, enrichment retentate on the filter membrane. The number of the filtering channels can be designed according to the requirement, and compared with the existing centrifugal separation method of the centrifugal machine, the number of centrifuge tubes capable of being placed in each centrifugation is limited.
Optionally, a liquid supply cavity located above the filter membrane is arranged on the upper connecting table, a lower channel located below the filter membrane is arranged on the lower connecting table, and the liquid supply cavity and the lower channel are combined to form the filtering channel.
Optionally, a groove located on the outer side of the filter membrane is arranged on the lower connecting platform, and a flange capable of being inserted into the groove is arranged on the upper connecting platform.
Optionally, a first sealing ring is arranged in the groove, and the flange is inserted into the groove to press the first sealing ring.
Optionally, the liquid supply device further comprises a liquid supply container, wherein a liquid outlet portion is arranged on the liquid supply container, and the liquid outlet portion is inserted into the liquid supply cavity.
Optionally, the filter membrane sealing device further comprises a second sealing ring, a limiting table located at the lower portion of the liquid supply cavity is arranged on the upper connecting table, and the second sealing ring is pressed on the edge of the filter membrane by the limiting table.
Optionally, at least one support table for supporting a filter membrane is arranged on the lower connecting table, and the support table is arranged on the inner wall of the lower channel.
Optionally, the upper connection table and the lower connection table are connected by bonding or snap-fit connection.
Optionally, the collection container is communicated with a negative pressure device.
Optionally, still include the fastener, the edge of connecting the platform is equipped with decurrent lower boss down, form draw-in groove down between the lateral wall of boss and lower connection platform down, the edge of going up the connection platform is equipped with ascending last boss, go up the boss with form the draw-in groove between the lateral wall of last connection platform, go up connection platform, down the connection platform laminating back, go up the shape that boss, lower boss combination formed with the recess phase-match of fastener, the fastener is followed the side of going up boss, lower boss is inserted, will go up boss, lower boss locking.
The invention also provides a microbial enzyme activity testing method, which comprises the following steps:
(1) preparing cell sap containing microorganisms to be detected, and performing suction filtration by adopting the filter device to obtain a filter membrane enriched with cells to be detected;
(2) providing a reaction solution containing a substrate, placing the filter membrane in the reaction solution, detecting the ratio of a product to the substrate after the reaction is finished, and calculating to obtain dehalogenation efficiency.
Alternatively, in the step (1), the volume of the cellular fluid pumped and filtered by each filtration channel is 2 to 10mL, specifically, 2mL, 3mL, 4mL, 5mL, 6mL, 7mL, 8mL, 9mL, 10mL, etc., preferably 3 to 9mL, further preferably 3 to 7mL, further preferably 4 to 6mL, further preferably 5 mL.
Optionally, in the step (1), the time required for suction filtration of the cell sap is 10 to 60s, specifically 10s, 20s, 30s, 40s, 50s, 60s, and the like, preferably 20 to 60s, further preferably 20 to 50s, further preferably 20 to 40s, further preferably 30 s.
Optionally, in the step (2), the ratio of the peak area of the substance of the product to the peak area of the substance of the substrate is calculated to obtain the dehalogenation efficiency.
Optionally, in the step (2), the product and the substrate are detected by a gas chromatography electron capture detector, and the ratio of the peak areas of the substances is calculated, i.e. the dehalogenation efficiency.
Optionally, in the step (2), the substrate is selected from polychlorinated biphenyl-180, and the product is polychlorinated biphenyl-153.
Optionally, in the step (2), the substrate concentration in the substrate-containing aqueous solution is 0.5 mg/L.
Alternatively, in step (2), each filter is independently placed in 0.4mL of the substrate-containing aqueous solution for reaction.
The invention also provides the application of the device in microbial enzyme activity test, cell collection and cell enzyme collection, and the device can also be used for collecting filtrate, and is preferably used for microbial enzyme activity test.
The invention has the following beneficial effects:
the invention provides a small-sized cell-enriching device with high efficiency aiming at the defects of low efficiency, complex operation, large consumption of consumable materials and the like of the existing cell-enriching device and method.
Drawings
FIG. 1 is a flow chart of a conventional microorganism cell in vitro enzyme activity test.
Fig. 2 is a schematic structural diagram of a filtering apparatus according to an embodiment of the present invention.
Fig. 3 is a cross-sectional view of the upper connecting stage in fig. 2.
Fig. 4 is a schematic cross-sectional view of the lower connecting station of fig. 2.
Fig. 5 is a schematic view of a filter device according to another embodiment of the present invention.
Fig. 6 is a schematic view of the combined filter device of fig. 5.
Fig. 7 is a schematic flow chart showing the filtering process using the filtering apparatus of fig. 6.
FIG. 8 is a graph showing the comparison of the reaction efficiency of dehalogenase pcbA1 after cell enrichment using the apparatus and method for testing enzyme activity of the present invention.
Description of numbering:
1-Collection mechanism
2-collecting container
21-liquid collecting Chamber
3-air vent
4-first seal ring
5-groove
6-Filter Membrane
7-liquid supply mechanism
8-Flange
9-second seal ring
10-liquid supply chamber
11-liquid supply container
110-liquid outlet part
12-lower channel
13-support bench
14-lower connecting table
15-upper connecting table
16-lower clamping table
161-lower card slot
17-go up calorie of platform
171-upper card slot
18-snap-on part
19-limiting table
Detailed Description
The invention is further described with reference to the drawings and the following detailed description, which are not intended to limit the invention in any way. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.
Unless otherwise indicated, reagents and materials used in the following examples are commercially available.
As shown in fig. 2-4, a filter device for testing microbial enzyme activity is provided, which comprises a liquid supply mechanism 7 for providing liquid to be filtered and a collection mechanism 1 for collecting filtrate, wherein the liquid supply mechanism 7 comprises an upper connection platform 15, the collection mechanism 1 comprises a collection container 2, a lower connection platform 14 for placing a filter membrane 6 is arranged on the collection container 2, the upper connection platform 15 and the lower connection platform 14 are detachably connected, the detachable connection structure is convenient to assemble into a closed structure during filtering, on the other hand, after filtering is finished, the upper connection platform 15 and the lower connection platform 14 are separated, the filter membrane 6 is taken out from the lower connection platform 14 for testing microbial enzyme activity, at least one filtering channel penetrates through the upper connection platform 15 and the lower connection platform 14, and the number of the filtering channels can be any number, such as 1, 2, 3,4, 5, 6, 7, 8, 9, 10 etc. each filter membrane corresponds a filtration passageway, and the quantity of seting up of filtration passageway is decided according to the needs of experiment, and the liquid that waits to filter in the confession liquid mechanism 7 passes filtration passageway and filter membrane 6, gets into collection liquid chamber 21 of collection mechanism 1, and the enrichment is held back on the filter membrane 6, and when the object of filtration was the cell sap, enrichment cell on the filter membrane 6 was the determinand.
In an embodiment, the upper connection table 15 is provided with a liquid supply chamber 10 located above the filter membrane 6, the lower connection table 14 is provided with a lower channel 12 located below the filter membrane 6, in some embodiments, the diameter of the lower channel 12 may be 10-12mm, specifically 10mm, 11mm, 12mm, and preferably 11mm, the liquid supply chamber 10 and the lower channel 12 form a filtration channel in combination, the liquid supply chamber 10 is located above the filter membrane 6, under the action of a vacuum pump, the liquid in the liquid supply container 11 passes through the liquid supply chamber 10, after being filtered by the filter membrane 6, the filtrate enters the liquid collection chamber 21, and the filter membrane 6 is enriched with the retentate. In some embodiments, the inner diameters of the liquid supply chamber 10 and the lower channel 12 may be kept uniform, and the lower channel 12 is positioned directly below the liquid supply chamber 10, so that the cellular liquid is sufficiently filtered.
In one embodiment, the lower connecting platform 14 is provided with a groove 5 positioned outside the filter membrane 6, the upper connecting platform 15 is provided with a flange 8 capable of being inserted into the groove 5, the groove 5 is internally provided with a first sealing ring 4, the flange 8 is inserted into the groove 5 to tightly press the first sealing ring 4, so that cell sap on the filter membrane 6 is fully separated during filtration, cells are intercepted on the filter membrane as much as possible, and the dehalogenation efficiency is further improved.
In one embodiment, the liquid supply device further comprises a liquid supply container 11, a liquid outlet portion 110 is arranged at the bottom of the liquid supply container 11, and the liquid outlet portion 110 is inserted into the liquid supply cavity 10, so that liquid in the liquid supply container 11 can smoothly flow into the liquid supply cavity 10, and a subsequent filtering step can be performed. The liquid supply container 11 may be an existing syringe, the bottom nozzle is the liquid outlet 110, after sucking a desired amount of cellular fluid through the piston handle, the bottom nozzle is inserted into the liquid supply chamber 10, and the vacuum pump is started to perform filtration.
In an embodiment, the device further comprises a second sealing ring 9, a limiting table 19 located at the lower portion of the liquid supply cavity 10 is arranged on the upper connecting table 15, and the second sealing ring 9 is pressed on the edge of the filter membrane 6 by the limiting table 19, so that the tightness of the device is effectively improved, and the filtering efficiency is improved.
In some embodiments, the liquid supply chamber 10 has a diameter of 4-6mm, and specifically may be 4mm, 4.5mm, 5mm, 5.5mm, 6mm, etc.
In an embodiment, a supporting platform 13 for supporting the filter membrane 6 is arranged on the lower connecting platform 14, the supporting platform 13 may be annular, and the middle of the supporting platform 13 is hollow, so that the filtrate can enter the liquid collecting cavity 21 from the filter membrane 6.
In an embodiment, the supporting platform 13 is disposed on the inner wall of the lower channel 12, and the supporting surface of the supporting platform 13 may be slightly lower than the upper surface of the lower connecting platform 14, so as to facilitate the placement of the filter membrane 6, thereby supporting the filter membrane 6 well, and facilitating the compression of the second sealing ring 9 to the edge of the filter membrane 6, thereby avoiding air leakage, and enabling the filtrate to flow into the liquid collecting chamber 21 through the lower channel 12 smoothly.
As shown in fig. 2, the liquid supply chamber 10 may be cylindrical, the inner diameter of the liquid supply chamber is slightly larger than the outer diameter of the liquid outlet 110, the liquid outlet 110 of the liquid supply container 11 is inserted into the liquid supply chamber 10, the vacuum pump is started, and the vacuum pump pumps away the air in the liquid collection chamber 21 and the liquid supply chamber 10 communicated with the liquid collection chamber through the vent hole 3, so that the liquid in the liquid supply container 11 flows to the filter membrane 6 to realize filtration.
In an embodiment, as shown in fig. 5 and 6, the bottom of the liquid supply chamber 10 may be in an inverted cone shape, after the liquid to be filtered is injected into the liquid supply chamber 10, the vacuum pump is turned on, and the liquid in the liquid supply chamber 10 flows toward the filter membrane 6 under the vacuum effect, so as to realize filtration.
In an embodiment, the liquid supply mechanism 7 and the collection mechanism 1 are connected by bonding, specifically, the upper connection platform 15 of the liquid supply mechanism 7 and the lower connection platform 14 of the collection mechanism 1 can be bonded together by glue to form a closed structure, so that cell sap can be filtered sufficiently, after the filtration is finished, the connection part is pried, the upper connection platform 15 can be detached from the lower connection platform 14, and then the filter membrane is taken out for subsequent enzyme activity test.
In another embodiment, the connection mode between the liquid supply mechanism 7 and the collection mechanism 1 is a snap connection, specifically, as shown in fig. 5 and 6, a downward lower boss 16 is provided at the edge of the lower connection platform 14, a lower slot 161 is formed between the lower boss 16 and the side wall of the lower connection platform 14, an upward upper boss 17 is provided at the edge of the upper connection platform 15, an upper slot 171 is formed between the upper boss 17 and the side wall of the upper connection platform 15, the upper boss 17 and the lower boss 16 are snap-connected by a snap fastener 18, the upper boss 17 and the lower boss 16 are provided at both left and right sides of the collection container 2, when the upper connection platform 15 and the lower connection platform 14 are attached, the upper boss 17 and the lower boss 16 are combined to form a T shape, the snap fastener 18 has a T-shaped groove, the lug of the snap fastener 18 is transversely inserted into the upper slot 171 and the lower slot 161, the upper boss 17 and the lower boss 16 are locked, and the upper connection platform 15, the lower connecting block 14 is locked. The shape formed by combining the upper boss 17 and the lower boss 16 may be another shape, and the shape of the engaging piece 18 is matched with the shape, so that the engaging piece 18 can be inserted from the side surface, and the upper boss 17 and the lower boss 16 can be smoothly locked.
The collecting container 2 is provided with a vent hole 3, the vent hole 3 is connected to a negative pressure device, and the negative pressure device can be a vacuum pump. The vent 3 is typically above the highest liquid level in the collection container 2 to avoid liquid being drawn into the conduit.
The vacuum pump is commercially available, and specifically may be model OL90A from danhao electromechanical devices limited, danyang, jiang su. The installation process of the device is as follows: place first sealing washer 4 in proper order on the recess 5 of lower joint table 14, place filter membrane 6 on a supporting bench 13, place second sealing washer 9 on filter membrane 6, will go up joint table 15 lock under on joint table 14, make flange 8 block in the recess 5, fix whole device with external fastener 18, make and go up joint table 15 and further compress tightly under on joint table 14, vacuum pump is connected to negative pressure device's air vent 3, pour 3-5mL cell sap into the cell sap container, supply in the liquid chamber 10 promptly, carry out the suction filtration, filtrate flows to the collection liquid chamber 21 of filtrating collecting container, the cell is held back on filter membrane 6. And (3) taking out the external clamping piece 18, separating the upper connecting platform 15 from the lower connecting platform 14, taking out the filter membrane 6, putting the bacteria-enriched filter membrane 6 into a reaction container, adding a reaction liquid, and carrying out enzyme catalysis reaction in a proper enzyme activity test environment.
The filtering device can be specifically made of PLA (polylactic acid), the polylactic acid has good thermal stability and processing temperature of 170-230 ℃, has good solvent resistance, and can be processed in various modes, such as extrusion, spinning, biaxial stretching, injection blow molding and the like. The product made of polylactic acid can be biodegraded, and has good biocompatibility, glossiness, transparency, hand feeling and heat resistance, and also has certain antibacterial property, flame retardance and ultraviolet resistance.
It was found through experiments that 95.68% of the substrate was converted to the product by only filtering 5mL of the cell liquid and reacting the filtered solid with 0.5mg/L of the substrate in a 0.4mL reaction solution.
In the following examples, the obligate anaerobic organic halide respiratory bacteria Dehalococcoides mccartyi CG1 are selected for experiments, the characteristics of other bacteria are similar to the characteristics of the obligate anaerobic organic halide respiratory bacteria, and the invention can be used for enzyme activity tests of other bacteria.
Example 1
In this example, an in vitro enzyme activity test of dehalogenase pcbA1 was performed using the obligate anaerobic organohalide respiring bacteria mccaratyi CG 1. The bacteria are obtained by separation and purification in the laboratory, and can be referred to as the following documents: genomics characterization of the three unique solutions of the catalyst systems of the previously described and previously described polycyclic polycarbonates, authors: shanquan Wang, Kern Rei Chun, Andrea Wilm et al, DOI: 10.1073/pnas.1404845111.
In this embodiment, the experimental apparatus for high-throughput microorganism in-vitro enzyme activity shown in fig. 5 to 6 is used, the operation flow is shown in fig. 7, the lower end of the cell sap container and the upper end of the filtrate collecting container are detachably connected together by a fastening mechanism, and the filter element (i.e., a filter membrane) is installed between the cell sap container and the filtrate collecting container. The external fixing mechanism is mutually embedded with flanges at two ends of the cell sap container and the filtrate collecting container, and the buckles are disassembled and assembled in a push-pull mode; injecting cell sap into the cell sap solution, connecting a connecting port of a filtrate collector with a negative pressure device, starting negative pressure, filtering filled liquid by a filtering element, and then flowing into the filtrate collector, wherein cells are retained on glass fibers; separating the cell sap container from the filtrate collector, namely detaching the glass fiber membrane, putting the glass fiber membrane into the container filled with the reaction solution, completely immersing the glass fiber membrane, and culturing for 48 hours at the temperature of 30 ℃ in a dark place. The filter element is a glass fiber filter membrane with the membrane aperture of 0.22 mu m and the diameter of 13mm, and is purchased from Yilong experimental equipment Limited of Ke jin. The pore size of the filter is determined according to the cell size in the experiment, and other pore sizes can be freely selected if other substances are filtered. The membrane diameter is determined by the design of the device, and in the present set of devices, it is preferred that each membrane diameter be the same size to fit the device.
Methods for preparing cellular fluids reference is made to the "Genomic characterization of the same, and" methods for obtaining and obtaining cellular fluids "(Shanquan waves, Kern Rei Chng, Andrea Wilm, Siyan Zhuao, Kun-Lin Yang, Niranjan Nagara, and Jianzhong. Deparatmeters of Civil and Environmental Engineering and Chemical and biological Engineering, National conversion of Singapore, Singapore 26; and comparative and Systems Biology, Genomic of Singapore 138672; Edied by Jaje M. tissue 353583, State of Marlateral and homogeneous of Martensile & 12108) section of the" materials and methods of the same, see section 138672, edition of Juniped M. Migap, Migap of Junipen, Juniper, Junipeditzeugee, Juniped 3683, and Juniped 3608, for obtaining materials ".
The cell fluid volume taken by each sample is 5mL, the maximum working pressure of the power of the used oil-free vacuum pump is-92 Kpa, the air extraction rate is 3.3L/s, the cell fluid needs 30s for complete filtration, 9 filtration channels are arranged on the filtration device, and 9 samples are filtered at one time.
A method for preparing a reaction solution containing a substrate is described in "Supporting Information" (Wang et al 10.1073/pnas 1404845111) in "Genomic characterization of hydrolytic solutions with high efficiency on a persistent polychlorinated biphenyls", specifically in "Enzymatic assays", wherein the concentration of the substrate in the reaction solution is 0.5mg/L, i.e., 0.5ppm, and the concentrations of the other components are the same as in the above-described reference, specifically, 100mM Tris. HCl (pH 7.0), 20mM methyl viologen, and 15mM titanium (III) citrate.
The name of dehalogenase pcbA1 is specifically disclosed in the literature: "Genomic characterization of a genetic deletion in a predetermined on a genetic multicated biphenyls".
The filter membrane does not need to be specially treated, the filter membrane only needs to be completely soaked into a reaction solution, the reaction substrate of dehalogenase pcbA1 is 2,2 ', 3,4,4 ', 5,5 ' -heptachlorobiphenyl (namely PCB-180, also called polychlorinated biphenyl-180), the using amount of the reaction solution is 0.4mL, the filter membrane is sealed at room temperature, the dehalogenation efficiency is calculated after 48-hour catalytic reaction, GC-ECD (gas chromatography electronic capture detector) is adopted to detect the ratio of a product to the substrate, and the reaction result is shown in figure 8, PCB-153 (namely polychlorinated biphenyl-153) is a product, and the dehalogenation efficiency is 95.68% (the original data is the peak area of a substance measured on a chromatograph, and the dehalogenation efficiency is calculated according to the peak area ratio of the product to the substrate). 2,2 ', 3,4,4 ', 5,5 ' -heptachlorobiphenyl was purchased from dr. ehrenstorfer GmbH, cat #: c20018000, Lot: G164639.
Methods for calculating dehalogenation efficiency references: a phylogenetic diagnostic of bacterial involved in segmented-free cultures (WangShanquan; He Jianzhong; Department of Civil and Environmental Engineering; National University of Singapore; Singapore);
published information in the literature is as follows:
JOURNAL:PloS one；
DOI:10.1371/journal.pone.0059178；
SOURCE is PubMed journal; DOI 10.1371/journal.bone.0059178; 2013 as YEAR; PAGES: e 59178;
PAGES:e59178；PUBLISHER:PubMed。
the literature: the present invention relates to a method for separating cellular fluid from cellular fluid by using an existing centrifugal tube, wherein the experimental result can be obtained from FIG. Fig.S4 (CG-12345-. The substrate, product and calculation method of the present example are different from the device used in the document, and the results show that the dehalogenation efficiency of the present example is as high as 95.68%, which is significantly higher than 78% in the prior art.
From the centrifugation method in the literature "genetic characterization of three times of microbial enzymes so as to obtain a microbial enzyme activity test method, it can be found that, in the existing microbial enzyme activity test method, a centrifuge is generally used for centrifugation, on one hand, the centrifuge can hold a limited number of centrifuge tubes each time, on the other hand, after each centrifugation, the cells enriched on the tube wall of the centrifuge tube are limited, on the other hand, each centrifugation needs 3-5 minutes, and after the supernatant is removed, the cell fluid needs to be added again, so that the cycle of centrifuging for multiple times is long, and the time is long. Even if the centrifugation is repeated, the dehalogenation efficiency measured finally is still far lower than that of the invention, and obviously, the number of cells which can be enriched is still limited by increasing the centrifugation times, and the invention successfully overcomes the defects, through a vacuum filtration mode, one-time filtration can be carried out without repeated operation, the enrichment efficiency is obviously higher than that of the prior art, and the method has the characteristic of high flux, moreover, the volume of the cell sap required for obtaining a sample by filtering is only about 2-10mL, the time required for filtering the cell sap is only 10-60s, the volume of the reaction liquid required is only 0.4mL, in the conventional centrifugation method, the volume of the cell fluid required for filtering a sample is about 1L, and the centrifugation is performed for twenty times, which takes about 1-2h, and the volume of the reaction solution is up to 4mL (the kind and molar amount of the substrate in the reaction solution of example 1 of the present invention are the same as those in the above-mentioned document).
In summary, the method has the following beneficial effects:
(1) the device can set the number of the filter channels as required, the cell sap containers are mutually independent and can be placed and taken simultaneously, repeated steps of one-by-one operation are omitted, and the use is more convenient;
(2) the method obviously shortens the time required by cell enrichment, simplifies the operation steps and reduces the consumption of experimental consumables while keeping high cell recovery rate;
(3) the invention greatly reduces the dosage of cell fluid and reaction solution while maintaining high reaction efficiency;
(4) the buckle mechanism of the device comprises flanges arranged at two ends of a cell liquid container and a filtrate collecting container and an external clamping piece, wherein the flanges at two ends of the cell liquid container and the filtrate collecting container are mutually embedded by the external clamping piece, and the buckle is disassembled and assembled in a push-pull mode, so that the cell liquid container and the filtrate collector can be tightly attached.
(5) The structure of the filtering element of the device is a sealing ring, a filtering membrane and a sealing ring, the filtering membrane is arranged between the two sealing rings, the filtering membrane is sealed tightly, the filtering membrane is made of glass fiber membranes and the like, the filtering membrane is not easy to deform in the suction filtration process, and cell liquid cannot leak along gaps between the device and the membrane, so that the cell recovery rate is improved.
(6) The filtrating collecting container of this device is equipped with the brace table that supports filtering element, and the recess of this brace table outer lane is equipped with the sealing washer, can guarantee that it is airtight in filtering process, and the filter effect is good.
(7) The cell sap container and the filtrate collecting container of the device can be made of PLA (polylactic acid), the polylactic acid has good thermal stability and processing temperature of 170-230 ℃, has good solvent resistance, and can be processed in various modes, such as extrusion, spinning, biaxial stretching and injection blow molding. The product made of polylactic acid can be biodegraded, and has good biocompatibility, glossiness, transparency, hand feeling and heat resistance, and also has certain antibacterial property, flame retardance and ultraviolet resistance.
(8) The filter membrane with enriched cells can directly perform catalytic reaction with reaction liquid containing a substrate.
(9) After the cell sap passes through the filter membrane, the cells can be completely retained on the filter membrane;
(10) compared with the traditional reaction system, the reaction system used by the device of the invention is reduced by about 10 times.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. The utility model provides a filter equipment for microbial enzyme test that lives, characterized in that, including the confession liquid mechanism (7) that is used for providing the liquid that waits to filter, be used for collecting collection mechanism (1) of filtrating, confession liquid mechanism (7) are including last joint table (15), collection mechanism (1) is including collecting container (2), be equipped with lower joint table (14) that are used for placing filter membrane (6) on collecting container (2), go up joint table (15), lower joint table (14) can dismantle the connection, go up joint table (15), run through on lower joint table (14) and be equipped with at least one filtration passageway, the liquid that waits to filter in the confession liquid mechanism (7) passes filtration passageway and filter membrane (6), get into collection liquid cavity (21) of collection mechanism (1), the enrichment retentate on filter membrane (6).
2. The filtration device of claim 1, wherein: the filter is characterized in that a liquid supply cavity (10) located above the filter membrane (6) is arranged on the upper connecting platform (15), a lower channel (12) located below the filter membrane (6) is arranged on the lower connecting platform (14), and the liquid supply cavity (10) and the lower channel (12) are combined to form the filter channel.
3. The filtration device of claim 1, wherein: the lower connecting platform (14) is provided with a groove (5) positioned on the outer side of the filter membrane (6), and the upper connecting platform (15) is provided with a flange (8) capable of being inserted into the groove (5).
4. A filter arrangement according to claim 3 wherein: a first sealing ring (4) is arranged in the groove (5), and the flange (8) is inserted into the groove (5) to compress the first sealing ring (4).
5. The filtration device of claim 2, wherein: the liquid supply device is characterized by further comprising a liquid supply container (10), wherein a liquid outlet part (110) is arranged on the liquid supply container (11), and the liquid outlet part (110) is inserted into the liquid supply cavity (10).
6. The filtration device of claim 1, wherein: the filter membrane sealing device is characterized by further comprising a second sealing ring (9), a limiting table (19) located at the lower portion of the liquid supply cavity (10) is arranged on the upper connecting table (15), and the second sealing ring (9) is pressed on the edge of the filter membrane (6) through the limiting table (19).
7. The filtration device of claim 2, wherein: the lower connecting table (14) is provided with at least one supporting table (13) for supporting the filter membrane (6), and the supporting table (13) is arranged on the inner wall of the lower channel (12).
8. The filtration device of claim 1, wherein: the upper connecting table (15) and the lower connecting table (14) are connected in an adhesive or clamping manner, and the collecting container (2) is communicated with a negative pressure device.
9. The filtration device of claim 1, wherein: still include fastener (18), the edge of lower joint chair (14) is equipped with decurrent lower boss (16), draw-in groove (161) down forms between the lateral wall of boss (16) and lower joint chair (14) down, the edge of connecting platform (15) is equipped with ascending last boss (17), go up boss (17) with form draw-in groove (171) between the lateral wall of last joint chair (15), go up joint chair (15), lower joint chair (14) laminating back, the shape that goes up boss (17), lower boss (16) combination and form with the recess phase-match of fastener (18), fastener (18) are followed the side of going up boss (17), lower boss (16) is inserted, will go up boss (17), lower boss (16) and lock.
10. A microbial enzyme activity test method is characterized by comprising the following steps:
(1) preparing a cell sap containing a microorganism to be detected, and performing suction filtration by using the filter device of any one of claims 1 to 9 to obtain a filter membrane enriched with cells to be detected;
(2) providing a reaction solution containing a substrate, placing the filter membrane in the reaction solution, detecting the ratio of a product to the substrate after the reaction is finished, and calculating to obtain dehalogenation efficiency.
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| CN201911018518.7A CN110938518A (en) | 2019-10-24 | 2019-10-24 | Filtering device and method for microbial enzyme activity test |
| PCT/CN2019/114518 WO2021077456A1 (en) | 2019-10-24 | 2019-10-31 | Filter device and method used for microbial enzymatic activity testing |
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| CN201911018518.7A CN110938518A (en) | 2019-10-24 | 2019-10-24 | Filtering device and method for microbial enzyme activity test |
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040009473A1 (en) * | 2002-06-07 | 2004-01-15 | Christopher Pease | Kit and process for microbiological for on-site examination of a liquid sample |
| CN1570075A (en) * | 2003-07-17 | 2005-01-26 | 友达光电股份有限公司 | Microbial cultivation sampling device |
| EP2305789A1 (en) * | 2008-06-27 | 2011-04-06 | Hitachi, Ltd. | Cartridge of microbial cell-capturing carrier, carrier treating device and method for counting microbial cells |
| CN103940660A (en) * | 2014-04-03 | 2014-07-23 | 华东理工大学 | Multichannel membrane enrichment device |
| WO2017055662A1 (en) * | 2015-10-01 | 2017-04-06 | Consejo Superior De Investigaciones Científicas (Csic) | Integrated filter-holder and microorganism concentration and detection method |
| WO2018018707A1 (en) * | 2016-07-26 | 2018-02-01 | 广州海力特生物科技有限公司 | Multi-layered centrifugal filter for exosome extraction |
| CN109012207A (en) * | 2018-08-21 | 2018-12-18 | 上海宸安生物科技有限公司 | A kind of cross-flow filtration system of circulating tumor cell enrichment |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN202263452U (en) * | 2011-08-12 | 2012-06-06 | 杭州吉沃科技有限公司 | Microbe and impurity filtering device |
| CN103131630A (en) * | 2011-11-28 | 2013-06-05 | 西安瑞捷生物科技有限公司 | Biological enzyme purification separating device |
| CN102495051A (en) * | 2011-12-01 | 2012-06-13 | 上海积彩医疗器械有限公司 | Device and method for quickly detecting biological activity and metabolism |
| CN202390438U (en) * | 2011-12-23 | 2012-08-22 | 潘茹茹 | Microbial limit filtering device |
| CN206624872U (en) * | 2017-04-06 | 2017-11-10 | 济南出入境检验检疫局检验检疫技术中心 | A kind of filter for microorganism detection |
| CN208949280U (en) * | 2018-10-11 | 2019-06-07 | 辽宁省分析科学研究院 | A kind of microbial limit filter device |
-
2019
- 2019-10-24 CN CN201911018518.7A patent/CN110938518A/en active Pending
- 2019-10-31 WO PCT/CN2019/114518 patent/WO2021077456A1/en active Application Filing
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040009473A1 (en) * | 2002-06-07 | 2004-01-15 | Christopher Pease | Kit and process for microbiological for on-site examination of a liquid sample |
| CN1570075A (en) * | 2003-07-17 | 2005-01-26 | 友达光电股份有限公司 | Microbial cultivation sampling device |
| EP2305789A1 (en) * | 2008-06-27 | 2011-04-06 | Hitachi, Ltd. | Cartridge of microbial cell-capturing carrier, carrier treating device and method for counting microbial cells |
| CN103940660A (en) * | 2014-04-03 | 2014-07-23 | 华东理工大学 | Multichannel membrane enrichment device |
| WO2017055662A1 (en) * | 2015-10-01 | 2017-04-06 | Consejo Superior De Investigaciones Científicas (Csic) | Integrated filter-holder and microorganism concentration and detection method |
| WO2018018707A1 (en) * | 2016-07-26 | 2018-02-01 | 广州海力特生物科技有限公司 | Multi-layered centrifugal filter for exosome extraction |
| CN109012207A (en) * | 2018-08-21 | 2018-12-18 | 上海宸安生物科技有限公司 | A kind of cross-flow filtration system of circulating tumor cell enrichment |
Non-Patent Citations (3)
| Title |
|---|
| SHANQUAN WANG ET AL.: ""Phylogenetically distinct bacteria involve extensive dechlorination of aroclor 1260 in sediment-free cultures"", 《PLOS ONE》 * |
| SHANQUAN WANGA ET AL.: ""Genomic characterization of three unique Dehalococcoides that respire on persistent polychlorinated biphenyls"", 《PNAS》 * |
| 吴梧桐主编: "《生物制药工艺学》", 30 April 2013 * |
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