CN116794139B - Method for rapidly screening plastic degradation microorganisms based on polymer membrane electrode sensing technology - Google Patents
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/333—Ion-selective electrodes or membranes
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/50—Reuse, recycling or recovery technologies
- Y02W30/62—Plastics recycling; Rubber recycling
Abstract
The invention discloses a method for rapidly screening microorganisms with plastic degradation activity. A method for quickly screening plastic degrading microbes based on polymer membrane electrode sensing technology is disclosed in more detail. The degradation capability of microorganisms on the plastic layer wrapped by the outer layer of the magnetic composite material is utilized to regulate and control the release flux of indicated ions in the composite material, and then the qualitative and/or quantitative detection of degradation efficiency of the microorganisms of the degraded plastic is realized through the response signals of the polymer membrane ion selective electrode on the composite material and the relationship between the indicated ion flux and the microbial degraded plastic; the composite material is formed by magnetic particles coated by plastic. Compared with the traditional method for screening microorganisms with plastic degradation activity, the method can be realized within a few hours. The invention provides a new idea for rapid screening of plastic degrading microorganisms.
Description
Technical Field
The invention discloses a method for rapidly screening microorganisms with plastic degradation activity. A method for quickly screening plastic degrading microbes based on polymer membrane electrode sensing technology is disclosed in more detail.
Background
The annual output of plastics is more than 3 hundred million tons, and the mass production and the use of plastic products and the unreasonable disposal and management mode of plastic waste make a large amount of plastic rubbish accumulate in the environment. Most plastics are difficult to degrade in a short time due to their chemically stable nature. The traditional plastic treatment method mainly comprises landfill, incineration and partial recovery treatment, and biodegradation is used as an emerging plastic treatment mode, and plastic waste can be degraded by biological enzymes or microorganisms, so that the method has the characteristics of environmental protection, low cost and the like, and has an application prospect in the aspect of solving the plastic pollution problem.
Some researchers have screened and separated a variety of microorganisms having the ability to degrade plastics from landfill sites, soil, ocean, and other environments. Existing methods available for assessing the ability of microorganisms to degrade plastics include: the method comprises the steps of observing the defect condition of the plastic surface in the plastic degradation process, by an electron microscopy method, an infrared spectrometry method for representing the change of functional groups on the plastic surface before and after plastic degradation, a chromatography method for measuring the change of molecular weight of the plastic polymer before and after plastic degradation, a weight loss method for measuring the change of mass before and after plastic degradation, and the like. However, the microorganisms existing in nature are various, and the degradation period of days or months is usually required by adopting the method, so that the experimental process is tedious and time-consuming. Therefore, how to quickly screen out microorganisms with high degradation capacity for specific plastics remains a great challenge.
Disclosure of Invention
Aiming at the defect of the existing method for rapidly screening microorganisms with plastic degradation activity, the invention aims to provide a method for rapidly screening plastic degradation microorganisms based on a polymer membrane electrode sensing technology.
In order to achieve the above purpose, the technical scheme of the invention is as follows:
a method for rapidly screening plastic degrading microorganisms based on a polymer membrane electrode sensing technology utilizes the degradation capability of microorganisms on a plastic layer wrapped by an outer layer of a magnetic composite material, regulates and controls the release flux of indicated ions in the composite material, and realizes qualitative and/or quantitative detection of degradation efficiency on degrading plastic microorganisms through the relationship between response signals of polymer membrane ion selective electrodes on the composite material, the indicated ion flux and the degrading plastic by the microorganisms; the composite material is formed by magnetic particles coated by plastic.
Further, the composite material and a sample to be detected are incubated together, if the sample contains degradation plastic microorganisms, the degradation plastic microorganisms degrade the plastic layer of the composite material, holes are formed on the surface of the composite material, medium conversion is carried out through a magnetic separation method, the composite material generates indication ion release flux under an acidic condition, and the polymer membrane ion selective electrode is caused to generate potential response signals, so that qualitative and/or quantitative detection of degradation efficiency of the degradation plastic microorganisms is realized.
The composite material and a sample to be detected are incubated in an inorganic salt culture medium at 25-35 ℃ for 1-12 hours.
The acidic condition is hydrochloric acid solution with a value of 1-5 pH.
The composite material has a three-layer structure, and the innermost layer is a magnetic substrate Fe 3 O 4 Particles which are convenient for realizing medium conversion by using a magnetic control separation method; the secondary outer layer is a signal indicating element Ca 3 (PO 4 ) 2 The coating layer is used for releasing calcium indicating ions after medium conversion; the outermost layer is a plastic coating layer of the identification element for identifying specific microorganisms of the degradable plastic.
The plastic is polystyrene, polyethylene, polypropylene, polyvinyl chloride, etc.
The application of the degrading microorganism screened by the method in degrading plastics.
The principle of the method for rapidly screening plastic degrading microorganisms is that the degradation effect of microorganisms on plastic layers is utilized to regulate and control the release flux of indicated ions (calcium), the relation between the potential signal of the polymer membrane ion selective electrode and the indicated ion flux and the efficiency of the plastic degrading microorganisms is established, and the rapid analysis of the efficiency of the plastic degrading microorganisms is realized.
The specific process of the method for rapidly screening plastic degrading microorganisms comprises the steps of bacterial strains and Fe 3 O 4 -Ca 3 (PO 4 ) 2 The magnetic composite material of plastic (polystyrene) is incubated in inorganic salt culture medium for several hours, the degradation of polystyrene plastic layer by bacterial strain is utilized to form holes or damage on the surface of the material, the medium is converted by magnetic separation method, the calcium indication ion flux in the magnetic composite material is released by acidic condition, and the polymer membrane calcium ion selective electrode is caused to generate potential response signal. The blank group contains Fe only 3 O 4 -Ca 3 (PO 4 ) 2 Inorganic salt culture medium of magnetic composite material of @ polystyrene.
By adopting the scheme, the invention has the beneficial effects that:
the present invention provides a method for rapidly screening microorganisms having plastic degrading activity, which uses Fe when the microorganisms are cultured under proper culture conditions 3 O 4 -Ca 3 (PO 4 ) 2 The polystyrene plastic coating in the magnetic composite material of the @ plastic (polystyrene) is degraded, the release flux of calcium indicating ions is regulated and controlled, and the rapid screening of plastic degrading microorganisms is realized within a few hours. The plastic layer in the magnetic composite material provided by the invention can be synthesized into different types according to experimental conditions, so that quick screening of microorganisms degrading different types of plastics is realized.
Drawings
Fig. 1 is a schematic diagram of a method for rapidly screening plastic degrading microorganisms according to an embodiment of the present invention.
FIG. 2 shows Fe provided in an embodiment of the present invention 3 O 4 、Fe 3 O 4 -Ca 3 (PO 4 ) 2 、Fe 3 O 4 -Ca 3 (PO 4 ) 2 SEM characterization of magnetic materials of polystyrene, wherein A is Fe 3 O 4 B is Fe 3 O 4 -Ca 3 (PO 4 ) 2 C is Fe 3 O 4 -Ca 3 (PO 4 ) 2 Polystyrene magnetic material.
FIG. 3 shows a blank and the known functions of degrading polystyrene plastics of Bacillus cereus strain and Fe 3 O 4 -Ca 3 (PO 4 ) 2 Potential response profile for 12 hours of co-incubation of polystyrene magnetic material.
FIG. 4 is a SEM morphology characterization of a strain with unknown function, wherein A is TXN-1 strain and B is TXN-2 strain.
FIG. 5 shows a blank and unknown functional strains and Fe 3 O 4 -Ca 3 (PO 4 ) 2 Potential response change plot of incubation of polystyrene magnetic material for 12 hours, wherein a is potential response change plot and B is potential change difference plot.
FIG. 6 is a SEM variation graph of TXN-1 and TXN-2 strains of unknown function incubated with polystyrene plastic film for 7-14 days; wherein A and B are SEM characterization images of polystyrene plastic film samples co-cultured with TXN-1 strain for 7 days and 14 days respectively, and the inset is SEM characterization image of blank control group cultured for 7 days and 14 days respectively; c and D are SEM characterization images of samples of polystyrene plastic films co-cultured with TXN-2 strain for 7 days and 14 days, respectively.
Detailed Description
The present invention is further described below with reference to examples, but the examples are only for helping to illustrate technical features of the present invention, and do not limit the scope of the present invention.
The invention is realized by synthesizing Fe 3 O 4 -Ca 3 (PO 4 ) 2 The @ polystyrene magnetic composite material is used for rapidly screening plastic degrading microorganisms, wherein a polystyrene plastic layer in the composite material is used as an identification element for identifying specific microorganisms; ca (Ca) 3 (PO 4 ) 2 The coating layer is used as a signal indicating element and is used for releasing calcium indicating ions after medium conversion; fe (Fe) 3 O 4 The microsphere is used as a magnetic substrate, so that the medium conversion is realized by using a magnetic separation method. The invention regulates and controls the release flux of the indicated ions by utilizing the degradation action of microorganisms on the plastic layer, establishes the relationship among the calcium ion selective electrode signal of the polymer membrane, the calcium indicated ion flux and the efficiency of degrading the plastic by the microorganisms, and realizes the rapid screening of the microorganisms with the plastic degradation activity. The blank group contains Fe only 3 O 4 -Ca 3 (PO 4 ) 2 Inorganic salt culture medium of magnetic composite material of @ polystyrene (as shown in figure 1).
Taking the method provided by the invention as an example for rapidly screening polystyrene plastic degrading microorganisms.
Example 1
Fe 3 O 4 -Ca 3 (PO 4 ) 2 Synthesis of magnetic composite material of @ polystyrene
（1）Fe 3 O 4 Synthesis of magnetic microspheres
Weigh 1.35 g FeCl 3 ·6H 2 O was dissolved in 40 mL glycol followed by the addition of 3.6 g anhydrous sodium acetate and 1.0 g polyethylene glycol. After the mixed solution was vigorously stirred at room temperature for 30 minutes, it was transferred to a high-pressure reaction vessel and reacted at 200℃for 8 h. After the reaction is finished, a black product Fe is obtained by separation through a magnetic separation method 3 O 4 The product was alternately washed several times with absolute ethanol and deionized water and dried in a vacuum oven at 60 c for 6 h to use.
（2）Fe 3 O 4 -Ca 3 (PO 4 ) 2 Synthesis of magnetic microspheres:
fe to be prepared 3 O 4 Magnetic microspheres (0.1 g) are uniformly dispersed in 100 mL deionized water under the ultrasonic condition, and 1 mL CaCl is added 2 (1.0M) solution was continued to sonicate 1. 1 h. Then 2 mL PBS buffer solution (0.1M, pH=7.4) is dripped under mechanical stirring, and stirring is continued for 4 h to obtain Fe 3 O 4 -Ca 3 (PO 4 ) 2 Magnetic microspheres. The product was washed several times with deionized water and stored at 4 ℃ for further use.
（3）Fe 3 O 4 -Ca 3 (PO 4 ) 2 Synthesis of magnetic composite material of @ polystyrene
Before synthesis, the styrene solution needs to be purified, namely, 20 mL styrene solution and 15 mL 1.0M NaOH solution are added into a 150 mL separating funnel and mixed thoroughly, the supernatant is left after standing and layering, and the above process is repeated three times to remove polymerization inhibitor in the styrene solution. The supernatant was then washed again 2 times with 30 mL deionized water to remove residual NaOH. Sequentially adding solvent (20 mL deionized water and 80 mL anhydrous ethanol mixed solution), purified styrene monomer (1 mL), cross-linking agent (0.1 mL, divinylbenzene) and emulsifying agent (0.3 g, polyvinylpyrrolidone) into a 250 mL three-neck flask, heating to 70deg.C under nitrogen protection to react 1 h, adding 3 mL initiator (2, 2-azobis (2-methylpropionamide) dihydrochloride aqueous solution, 1.7 wt%) to continue reaction for 10 min, and adding 20 mL Fe prepared above 3 O 4 -Ca 3 (PO 4 ) 2 Dispersion of 70 ° The reaction was continued for 48 hours at C. Coating the obtained polystyrene with Fe by using a magnetic separation method 3 O 4 -Ca 3 (PO 4 ) 2 Washing the microspheres several times in deionized water to remove residual reactants and free polystyrene nanoparticles, and subjecting the product to60. Drying 6 h in a vacuum drying oven at the temperature for later use.
According to the method, the polystyrene is replaced by polyethylene, polypropylene, polyvinyl chloride and the like. Obtaining different kinds of plastic layers coated with Fe 3 O 4 -Ca 3 (PO 4 ) 2 The magnetic composite material of (2) can be prepared by changing corresponding experimental conditions through the synthesis method.
The Scanning Electron Microscope (SEM) characterization of the composite material is shown in FIG. 2, comparing unmodified Fe 3 O 4 Microsphere (A), ca 3 (PO 4 ) 2 After formation of the ion-indicating coating layer (B), the coating layer is formed of Fe 3 O 4 Irregularly shaped fragments can be observed on the microsphere surface. After the polystyrene plastic coating layer is formed by continuous polymerization, the surface morphology of the microsphere is obviously changed and becomes smooth, which indicates that the material is successfully synthesized.
Example 2
Verification of degradation of plastics Using the composite obtained in example 1 above
The microorganism enrichment medium used was 2216E liquid medium (available from Qingdao sea Bo Biotechnology Co., ltd.). Composition (g/L) of inorganic salt medium for evaluating plastic degrading ability of microorganism: 0.92 g K 2 HPO 4 ·3H 2 O，0.7 g KH 2 PO 4 ，0.7 g MgSO 4 ·7H 2 O，1.0 g NH 4 NO 3 ，0.005 g NaCl，0.002 g FeSO 4 ·7H 2 O，0.002 g ZnSO 4 ·7H 2 O and 0.001 g MnSO 4 The preparation is sterilized in a sterilizing pot at 121 ℃ for 20 minutes before use.
Referring to fig. 1, the composite material obtained in example 1 above was incubated with bacillus cereus strain ATCC14579 (Bacillus cereus CH 6) known to have the ability to degrade polystyrene plastics for a period of time, and the concentration of calcium ions released from the magnetic composite material into a dilute hydrochloric acid solution was detected using a polymer membrane calcium ion selective electrode to verify the feasibility of the method.
Meanwhile, the composite material obtained in example 1 was used as a control.
The method comprises inoculating Bacillus cereus CH strain in logarithmic phase of growth with 10% of inoculating amount in 20 mL sterile inorganic salt culture medium and Fe 3 O 4 -Ca 3 (PO 4 ) 2 Magnetic composites of polystyrene (2 mg/mL) and incubated at 33℃in a shaker at 90 rpm. The blank control group is 20 mL inorganic salt culture medium without inoculating bacteria and Fe 3 O 4 -Ca 3 (PO 4 ) 2 Magnetic composites of polystyrene (2 mg/mL), other experimental conditions were kept consistent. After culturing for 12 hours, adopting a magnetic control separation method to separate Fe 3 O 4 -Ca 3 (PO 4 ) 2 Separating the magnetic composite material of the @ polystyrene from the inorganic salt culture medium, washing the magnetic composite material with deionized water for a plurality of times, then redispersing the magnetic composite material in 10 mL dilute hydrochloric acid (pH=3) solution, oscillating for 5 min, and then separating out Fe again 3 O 4 -Ca 3 (PO 4 ) 2 The concentration of calcium ions released from the magnetic composite into the dilute hydrochloric acid solution is detected by using a polymer membrane calcium ion selective electrode. The above experiments were performed in triplicate.
The test results are shown in FIG. 3, which shows that Fe was inoculated with Bacillus cereus CH6 strain compared with the blank group 3 O 4 -Ca 3 (PO 4 ) 2 The potential signal generated in the magnetic composite material of @ polystyrene was greater, indicating that strain Bacillus cereus CH6 caused a change in electrode potential by degrading the polystyrene plastic layer of the magnetic composite material, releasing calcium-indicating ions into the solution. This result demonstrates the feasibility of a method for rapidly screening plastics degrading microorganisms based on polymer membrane electrode sensing technology.
Example 3
The ability of the microbiologically degradable plastic was evaluated based on polymer membrane electrode sensing technology:
the sediment of the cold spring liquid on the seafloor of the southwest part of taiwan is taken, and the capability of degrading polystyrene plastic is evaluated by a polymer membrane electrode sensing technology.
The acquisition process comprises the following steps: the subsea cold spring liquid sediment supernatant (22 DEG 06'55.457' '-26.831' '-N, 119 DEG 17'07.302'' -10.153'' -E, depth about 1100 m) was enriched with sterile polystyrene plastic film in mineral salt medium containing a small amount of nutrients at 33 ℃ for 30 days. And then separating and purifying microorganisms attached to the surface of the polystyrene plastic film, and continuously subculturing to realize mass enrichment of strains. And finally, identifying the separated and purified strain by a 16S rRNA gene sequencing technology.
Two strains of enterobacteria of unknown function were selected, designated TXN-1 and TXN-2, respectively, and the morphology of the TXN-1 and TXN-2 strains of unknown function used was observed by SEM. The TXN-1 strain (FIG. 4A) was in the form of a rod, and the cells were elongated, straight or slightly bent, and no hypha was observed around the cells; the TXN-2 strain (FIG. 4B) is similar to the TXN-1 strain in morphology, and is in a rod shape, and the cells are elongated, straight or slightly curved, but differ in that the cell surface carries hyphae (black arrow indication).
Two unknown TXN-1 and TXN-2 strains were inoculated into 2216E liquid medium, respectively, and placed in a shaker at 33℃for overnight culture, and the next day was re-inoculated into a new 2216E liquid medium for continuous culture for 3-5 hours to the growth log phase.
Taking Fe prepared in example 1 above 3 O 4 -Ca 3 (PO 4 ) 2 Polystyrene magnetic composite (2 mg/mL) was added to inorganic salt medium inoculated with 10% of TXN-1 and TXN-2 strains in log phase, respectively, and incubated at 33℃for 12 hours in a shaker at 90 rpm. The blank control group is inorganic salt culture medium without bacteria and Fe 3 O 4 -Ca 3 (PO 4 ) 2 Magnetic composite material of @ polystyrene, and keeping other experimental conditions consistent. After culturing for 12 hours, adopting a magnetic control separation method to separate Fe 3 O 4 -Ca 3 (PO 4 ) 2 Separating the magnetic composite material of the @ polystyrene from the inorganic salt culture medium, washing the magnetic composite material with deionized water for a plurality of times, then redispersing the magnetic composite material in 10 mL dilute hydrochloric acid (pH=3) solution, oscillating for 5 min, and then separating out Fe again 3 O 4 -Ca 3 (PO 4 ) 2 Polyphenyl @ polystyreneEthylene magnetic composite material.
Further, the concentration of calcium ions released from the magnetic composite into the dilute hydrochloric acid solution was then measured using a polymer membrane calcium ion selective electrode, and the experiment was performed in triplicate.
The preparation process of the polymer film calcium ion selective electrode comprises the following steps: 8.28 and mg of ionophore ETH 129,7.92 mg of ion exchanger sodium tetrakis (3, 5-bis (trifluoromethyl) phenyl) borate, 229.32 mg of plasticizer o-nitrophenyl octyl ether and 114.48 mg of film matrix polyvinyl chloride are respectively weighed into a dry and transparent glass bottle, and are fully stirred for 2 hours after 3.6 and mL of tetrahydrofuran solution is added to be fully dissolved. Pouring the mixture into a glass ring with the diameter of 3.6 and cm at room temperature, and standing overnight in a constant temperature and humidity box until tetrahydrofuran is completely volatilized to obtain the transparent and elastic calcium ion selective sensitive film with the thickness of about 200 mu m. Cutting the calcium ion selective sensitive film into small discs with the diameter of about 6.0 and mm by using a puncher, and adhering the small discs to the tail end of a polyvinyl chloride pipe by using tetrahydrofuran to obtain the polymer film calcium ion selective electrode. Placing in a constant temperature and humidity box for about 2 hours until tetrahydrofuran is completely volatilized, and injecting 10 into the electrode cavity -3 M CaCl 2 Is used as an internal liquid to be placed at 10 -3 M CaCl 2 The solution was used after activation overnight.
Fe is regulated and controlled by utilizing degradation of microorganisms to plastics 3 O 4 -Ca 3 (PO 4 ) 2 The release flux of the indication ions in the magnetic polystyrene composite material is combined with the polymer membrane ion selective electrode, so that the rapid detection of the efficiency of the microbial degradation plastic is realized. As shown in the graph of FIG. 5, the concentration of calcium ions released from the magnetic composite material after 12 hours of co-culture with TXN-2 strain causes a significant electrode potential signal, which indicates that the degradation capability of the magnetic composite material on a polystyrene plastic layer is stronger; the electrode potential signal caused by the concentration of released calcium ions in the magnetic composite material after being co-cultured with the TXN-1 strain is minimum, which shows that the degradation capability of the magnetic composite material on a polystyrene plastic layer is the weakest.
The unknown strain used above was evaluated for its ability to degrade plastics by conventional SEM method
TXN-1 and TXN-2 strains were inoculated into 2216E liquid medium and placed in a shaker at 33℃overnight, and the next day was re-inoculated into a new 2216E liquid medium and continued to culture for 3-5 hours until the logarithmic phase of growth. The two strains (TXN-1, TXN-2) in log phase of growth were again inoculated in 10% inoculum size into 50 mL sterile Erlenmeyer flasks containing 20 mL sterilized inorganic salt medium and sterilized polystyrene plastic film samples and placed in a constant temperature and humidity cabinet at 33℃for cultivation. As a control, the unvaccinated inorganic salt medium and the polystyrene plastic film sample were cultured under the same conditions. After 7 days and 14 days of culture, samples of the polystyrene plastic film were taken out, microbial films formed on the surfaces thereof were removed by ultrasonic cleaning, and degradation of the polystyrene plastic was observed under SEM. The above experiments were performed in triplicate.
Under the above experimental conditions, degradation of the polystyrene plastic film samples after 7 days and 14 days of culture with TXN-1 and TXN-2 in an inorganic salt medium was observed by SEM, and the results are shown in FIG. 6. Wherein A is an SEM characterization of a polystyrene plastic film sample co-cultured with TXN-1 strain for 7 days, B is an SEM characterization of a polystyrene plastic film sample co-cultured with TXN-1 strain for 14 days, and the inset is an SEM characterization of a blank control group cultured for 7 days and 14 days; c and D are SEM characterization images of samples of polystyrene plastic films co-cultured with TXN-2 strain for 7 days and 14 days, respectively. Compared with a blank control group, under the condition that no carbon source exists in an inorganic salt culture medium, folds, ravines and pits with different degrees appear on the surface of a polystyrene plastic film sample degraded by different bacteria, and according to the surface morphology change degree, the degradation capability of different bacteria on the polystyrene plastic film can be obtained as follows: TXN-2 > TXN-1. This result is consistent with that obtained using the polymer membrane electrode sensing technique.
The previous description of the embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above-described embodiments. Those skilled in the art will appreciate that, in light of the principles of the present invention, improvements and modifications can be made without departing from the scope of the invention.
Claims (6)
1. A method for rapidly screening plastic degrading microorganisms based on a polymer membrane electrode sensing technology is characterized in that the degradation capability of microorganisms on a plastic layer wrapped by a magnetic composite material outer layer is utilized to regulate and control the release flux of indicated ions in the composite material, and then the qualitative and/or quantitative detection of the microbial degradation effect of the degrading plastic is realized through the response signals of polymer membrane ion selective electrodes on the composite material, the relationship between the indicated ion flux and the microbial degradation plastic; the composite material is formed by magnetic particles coated by plastic.
2. The method for rapidly screening plastic degrading microorganisms based on the polymer membrane electrode sensing technology according to claim 1, wherein the composite material is incubated with a sample to be detected, if the sample contains degrading plastic microorganisms, the plastic layer of the composite material is degraded, holes are formed on the surface of the composite material, medium conversion is carried out by a magnetic separation method, the composite material generates indication ion release flux under an acidic condition, and the polymer membrane ion selective electrode is caused to generate potential response signals, so that qualitative and/or quantitative detection of the degradation effect of the degrading plastic microorganisms is realized.
3. A method for rapid screening of plastics degrading micro-organisms based on polymer membrane electrode sensing technology according to claim 1 or 2, wherein the composite material is incubated with the sample to be tested in an inorganic salt medium at 25-35 ℃ for 1-12 hours.
4. The method for rapid screening of plastic degrading microorganisms based on polymer membrane electrode sensing technology according to claim 2, wherein the acidic condition is 1-5 pH value hydrochloric acid solution.
5. The method for rapidly screening plastic degrading microorganisms based on polymer membrane electrode sensing technology as claimed in claim 1 or 2, wherein the composite material has a three-layer structure, and the innermost layer is a magnetic substrate Fe 3 O 4 Particles which are convenient for realizing medium conversion by using a magnetic control separation method; the secondary outer layer is a signal indicating element Ca 3 (PO 4 ) 2 The coating layer is used for releasing calcium indicating ions after medium conversion; the outermost layer is a plastic coating layer of the identification element for identifying specific microorganisms of the degradable plastic.
6. The method for rapid screening of plastic degrading microorganisms based on polymer membrane electrode sensing technology according to claim 5, wherein the plastic is polystyrene, polyethylene, polypropylene or polyvinyl chloride.
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