CN113750437A - Method for enhancing interface activation to efficiently biodegrade PET (polyethylene terephthalate) - Google Patents
Method for enhancing interface activation to efficiently biodegrade PET (polyethylene terephthalate) Download PDFInfo
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- CN113750437A CN113750437A CN202111112786.2A CN202111112786A CN113750437A CN 113750437 A CN113750437 A CN 113750437A CN 202111112786 A CN202111112786 A CN 202111112786A CN 113750437 A CN113750437 A CN 113750437A
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- 230000004913 activation Effects 0.000 title claims abstract description 19
- 238000000034 method Methods 0.000 title claims abstract description 18
- 239000005020 polyethylene terephthalate Substances 0.000 title description 61
- 229920000139 polyethylene terephthalate Polymers 0.000 title description 60
- -1 polyethylene terephthalate Polymers 0.000 title description 8
- 230000002708 enhancing effect Effects 0.000 title description 2
- 238000006731 degradation reaction Methods 0.000 claims abstract description 23
- 230000015556 catabolic process Effects 0.000 claims abstract description 21
- 229920000426 Microplastic Polymers 0.000 claims abstract description 18
- 239000012190 activator Substances 0.000 claims abstract description 15
- 239000007788 liquid Substances 0.000 claims abstract 3
- 239000001963 growth medium Substances 0.000 claims description 27
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- 229920000728 polyester Polymers 0.000 claims description 14
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 claims description 12
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 12
- JIAARYAFYJHUJI-UHFFFAOYSA-L zinc dichloride Chemical compound [Cl-].[Cl-].[Zn+2] JIAARYAFYJHUJI-UHFFFAOYSA-L 0.000 claims description 12
- 230000001954 sterilising effect Effects 0.000 claims description 11
- 239000000126 substance Substances 0.000 claims description 8
- 235000019270 ammonium chloride Nutrition 0.000 claims description 6
- 239000011609 ammonium molybdate Substances 0.000 claims description 6
- 235000018660 ammonium molybdate Nutrition 0.000 claims description 6
- APUPEJJSWDHEBO-UHFFFAOYSA-P ammonium molybdate Chemical compound [NH4+].[NH4+].[O-][Mo]([O-])(=O)=O APUPEJJSWDHEBO-UHFFFAOYSA-P 0.000 claims description 6
- 229940010552 ammonium molybdate Drugs 0.000 claims description 6
- 238000006065 biodegradation reaction Methods 0.000 claims description 6
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims description 6
- 239000004327 boric acid Substances 0.000 claims description 6
- JZCCFEFSEZPSOG-UHFFFAOYSA-L copper(II) sulfate pentahydrate Chemical compound O.O.O.O.O.[Cu+2].[O-]S([O-])(=O)=O JZCCFEFSEZPSOG-UHFFFAOYSA-L 0.000 claims description 6
- 229940044631 ferric chloride hexahydrate Drugs 0.000 claims description 6
- 239000004519 grease Substances 0.000 claims description 6
- NQXWGWZJXJUMQB-UHFFFAOYSA-K iron trichloride hexahydrate Chemical compound O.O.O.O.O.O.[Cl-].Cl[Fe+]Cl NQXWGWZJXJUMQB-UHFFFAOYSA-K 0.000 claims description 6
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims description 6
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 claims description 6
- SCVOEYLBXCPATR-UHFFFAOYSA-L manganese(II) sulfate pentahydrate Chemical compound O.O.O.O.O.[Mn+2].[O-]S([O-])(=O)=O SCVOEYLBXCPATR-UHFFFAOYSA-L 0.000 claims description 6
- 239000001103 potassium chloride Substances 0.000 claims description 6
- 235000011164 potassium chloride Nutrition 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- 238000002360 preparation method Methods 0.000 claims description 6
- 239000011780 sodium chloride Substances 0.000 claims description 6
- 238000004659 sterilization and disinfection Methods 0.000 claims description 6
- 239000000758 substrate Substances 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 239000011592 zinc chloride Substances 0.000 claims description 6
- 235000005074 zinc chloride Nutrition 0.000 claims description 6
- 239000002609 medium Substances 0.000 claims 3
- 101150015738 Fev gene Proteins 0.000 claims 1
- 229920001410 Microfiber Polymers 0.000 claims 1
- 102100037681 Protein FEV Human genes 0.000 claims 1
- 230000001580 bacterial effect Effects 0.000 claims 1
- 239000003658 microfiber Substances 0.000 claims 1
- 238000005728 strengthening Methods 0.000 claims 1
- 102000004190 Enzymes Human genes 0.000 abstract description 11
- 108090000790 Enzymes Proteins 0.000 abstract description 11
- 239000003513 alkali Substances 0.000 abstract description 10
- 239000011942 biocatalyst Substances 0.000 abstract description 5
- 230000000593 degrading effect Effects 0.000 abstract description 4
- 230000003197 catalytic effect Effects 0.000 abstract description 2
- 229920000136 polysorbate Polymers 0.000 abstract description 2
- 238000002144 chemical decomposition reaction Methods 0.000 abstract 2
- 231100000252 nontoxic Toxicity 0.000 abstract 1
- 230000003000 nontoxic effect Effects 0.000 abstract 1
- 239000002861 polymer material Substances 0.000 abstract 1
- 238000009630 liquid culture Methods 0.000 description 16
- 239000000835 fiber Substances 0.000 description 10
- 229920003023 plastic Polymers 0.000 description 7
- 239000004033 plastic Substances 0.000 description 7
- 238000001035 drying Methods 0.000 description 6
- 241000894006 Bacteria Species 0.000 description 5
- 241000589518 Comamonas testosteroni Species 0.000 description 5
- 238000001816 cooling Methods 0.000 description 5
- 238000012258 culturing Methods 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- 239000013543 active substance Substances 0.000 description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 229920001213 Polysorbate 20 Polymers 0.000 description 2
- MUMGGOZAMZWBJJ-DYKIIFRCSA-N Testostosterone Chemical compound O=C1CC[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 MUMGGOZAMZWBJJ-DYKIIFRCSA-N 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 239000003054 catalyst Substances 0.000 description 2
- 238000004043 dyeing Methods 0.000 description 2
- 238000003912 environmental pollution Methods 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 238000010406 interfacial reaction Methods 0.000 description 2
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 2
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 2
- 241000589519 Comamonas Species 0.000 description 1
- 229920001214 Polysorbate 60 Polymers 0.000 description 1
- 229920004933 Terylene® Polymers 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 230000005661 hydrophobic surface Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 229960003604 testosterone Drugs 0.000 description 1
- 239000004753 textile Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D3/00—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances
- A62D3/02—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances by biological methods, i.e. processes using enzymes or microorganisms
-
- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D2101/00—Harmful chemical substances made harmless, or less harmful, by effecting chemical change
- A62D2101/20—Organic substances
- A62D2101/28—Organic substances containing oxygen, sulfur, selenium or tellurium, i.e. chalcogen
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- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Toxicology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Business, Economics & Management (AREA)
- Emergency Management (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明基于耐碱全细胞生物催化剂，利用一种界面活化剂创建了降解PET微塑料的化学‑生物联合催化体系。该方法操作步骤如下：首先按配方配制碱性液体培养基；然后，另外添加吐温系列界面活化剂，置于超声仪器中充分分散，在恒温摇床上进行界面活化2h；最后，将体系灭菌，接入耐碱菌株进行生物‑化学联合降解。该方法操作简单、绿色环保、无毒害、成本低廉，能高效地将降解界面活化，降低PET表面活化能，从而提高PET微塑料的生物‑化学联合降解效率。本发明不仅为PET微塑料的安全、彻底去除提供了有效的解决方案，还为实现其他难降解人造高分子材料的绿色处理树立了示范和引领作用。Based on the alkali-resistant whole-cell biocatalyst, the invention utilizes an interfacial activator to create a chemical-biological joint catalytic system for degrading PET microplastics. The operation steps of the method are as follows: firstly, the alkaline liquid medium is prepared according to the formula; then, the Tween series interface activator is additionally added, placed in an ultrasonic instrument to fully disperse, and the interface is activated on a constant temperature shaker for 2 hours; finally, the system is sterilized , access to alkali-tolerant strains for combined bio-chemical degradation. The method is simple to operate, environmentally friendly, non-toxic, and low-cost, and can efficiently activate the degradation interface and reduce the surface activation energy of PET, thereby improving the combined bio-chemical degradation efficiency of PET microplastics. The present invention not only provides an effective solution for the safe and complete removal of PET microplastics, but also sets a demonstration and leading role for realizing the green treatment of other refractory artificial polymer materials.
Description
Technical Field
The invention relates to a polyester degradation treatment method, in particular to a method for catalyzing efficient degradation of PET by an interfacial biological activator, and belongs to the field of polyester textile processing.
Technical Field
Biodegradation is considered to be the safest and most thorough method for solving the problems of the terylene micro plastic. However, PET is a bio-resistant high molecular compound having high crystallinity, a dense structure and a highly hydrophobic surface. In addition, as a chemically synthesized material, polyester appears for a short time as compared with a natural product, and microorganisms capable of efficiently decomposing PET have not been developed in nature.
Although, various microorganisms and enzymes for biodegradation of PET have been reported in recent years, their productivity is limited. Considering that the degradation of PET is an interfacial catalytic reaction, the use of an interfacial catalyst is considered to be an effective method for improving the degradation of PET. The interfacial catalyst, whether chemically or biologically degraded, can reduce surface tension, making enzymes and chemical molecules more likely to contact the polymer surface, and ultimately increasing the efficiency of the interfacial reaction.
The invention relates to a chemical-biological combined catalytic system for degrading PET (polyethylene terephthalate) micro-plastic by utilizing an interfacial activator based on an alkali-resistant whole-cell biocatalyst. In the process of corroding the PET surface by enzyme and alkaline molecules, the interfacial biological reaction is enhanced by applying an interfacial activator, and the biodegradation efficiency is enhanced. Because the interfacial activator used in the invention belongs to esters, the interfacial activator can also be used as a carbon source to be utilized by microorganism growth and reproduction in the PET biodegradation process. Therefore, the method can not further cause environmental pollution and meets the requirements of ecological dyeing and finishing.
Disclosure of Invention
The invention aims to solve the technical problem of providing a novel method for efficiently degrading PET by utilizing interface activation based on an alkali-resistant whole-cell biocatalyst aiming at the defect of low efficiency of the existing enzyme and microorganism degradation of PET.
The technical scheme adopted by the invention is as follows: a novel method for efficiently degrading PET by utilizing interface activation based on an alkali-resistant whole-cell biocatalyst. The method is characterized by comprising the following steps:
(1) preparation of PET micro plastic: firstly, polyester superfine fiber is refined, surface grease is removed, and the polyester superfine fiber is fully dried after being washed for multiple times; then, crushing the substrate to the size of micro plastic, collecting the screened PET powder, washing with water, and drying for later use.
(2) Interface activation: preparing an alkaline liquid culture medium according to a culture formula of the alkali-resistant Comamonas testosteroni, and additionally adding 0.5-3% of interfacial activator by volume. The system is placed in an ultrasonic instrument to ensure that the PET and the interfacial activator are fully and uniformly dispersed, and the PET and the interfacial activator are placed on a constant temperature shaking table at 37 ℃ and 140rpm for treatment for 2 to 5 hours to ensure that the PET surface is fully emulsified. The formula of the culture medium is as follows: 1.00g/L of ammonium chloride, 1.4g/L of sodium hydroxide, 3.7g/L of potassium chloride, 0.50g/L of sodium chloride, 0.25g/L of magnesium sulfate heptahydrate, 1g/L of PET, 0.5 mu g/L of boric acid, 0.2 mu g/L of ferric chloride hexahydrate, 0.4 mu g/L of manganese sulfate pentahydrate, 0.4 mu g/L of zinc chloride, 40.0 mu g/L of copper sulfate pentahydrate and 0.2 mu g/L of ammonium molybdate.
(3) Biological-chemical combined degradation of PET (polyethylene terephthalate) micro-plastic: taking out the culture medium in the step (2), sterilizing at high temperature and high pressure for 20min, cooling to room temperature in a super clean bench, transferring into 5% bacteria solution, and culturing on a constant temperature shaking table at 37 ℃ and 140rpm for 3-5 days.
(4) And (4) preparing an alkaline liquid culture medium again, replacing 20-40% of the alkaline liquid culture medium into the degradation system in the step (3) after interface activation and sterilization, and continuously placing the alkaline liquid culture medium into a constant-temperature incubator at 37 ℃ and 140rpm for culture. This was repeated for 60 days.
Preferably, the drying method of the PET after refining in the step (1) is: and (5) freeze drying.
Preferably, the way of pulverizing PET in step (1) is: physical cutting, using an ultracentrifugal cutter.
Preferably, in the step (2), the ultrasonic temperature is 30 ℃, the ultrasonic frequency is 40KHz, and the ultrasonic time is 0.5 h; the surfactant is Tween 20, the volume is 1%, and the interfacial activation time is 2 h.
Preferably, the displacement time of the new fermentation broth in step (3) is 3 days.
Preferably, the displaced volume of fresh fermentation broth in step (4) is 30%.
Due to the adoption of the technical scheme, the method has the following beneficial effects:
(1) degradation of PET is an interfacial reaction, both chemical and biological. Therefore, the addition of the interfacial activator can enable the alkaline molecules and the extracellular enzyme to be quickly and specifically combined with the PET micro-plastic, and promote the high-efficiency degradation reaction.
(2) From the aspects of environmental protection and high-efficiency degradation, the invention utilizes the alkali-resistant testosterone comamonas as the whole-cell biocatalyst, not only can lead PET to be slightly degraded in a weak alkaline environment, but also saves the steps of enzyme extraction and purification in the PET biodegradation process.
(3) Because the added interface activator is an ester molecule, the interface activator can be assimilated and absorbed by microorganisms as a carbon source in the degradation process of PET. Therefore, the method does not cause environmental pollution, accords with the development direction of ecological dyeing and finishing, and has wide application prospect.
Detailed Description
Example 1
(1) Preparation of PET micro plastic: firstly, polyester superfine fibers and slices are refined, surface grease is removed, and the polyester superfine fibers and the slices are fully dried after being washed for multiple times; then, crushing the substrate to the size of micro plastic, collecting the screened PET powder, washing with water, and drying for later use.
(2) Interface activation: preparing 200ml of culture solution according to a culture formula of alkali-resistant Comamonas testosteroni in a 500ml conical flask, and adding 1% by volume of Tween 20; the system is placed in an ultrasonic instrument to ensure that the PET and the interfacial active agent are uniformly dispersed, and the system is placed on a constant temperature shaking table at 37 ℃ and 140rpm for treatment for 2 to 5 hours to ensure that the surface of the PET is fully emulsified. The formula of the culture medium is as follows: 1.00g/L of ammonium chloride, 1.4g/L of sodium hydroxide, 3.7g/L of potassium chloride, 0.50g/L of sodium chloride, 0.25g/L of magnesium sulfate heptahydrate, 1g/L of PET, 0.5 mu g/L of boric acid, 0.2 mu g/L of ferric chloride hexahydrate, 0.4 mu g/L of manganese sulfate pentahydrate, 0.4 mu g/L of zinc chloride, 40.0 mu g/L of copper sulfate pentahydrate and 0.2 mu g/L of ammonium molybdate.
(3) Biological-chemical combined degradation of PET (polyethylene terephthalate) micro-plastic: taking out the culture medium in the step (2), sterilizing at high temperature and high pressure for 20min, cooling to room temperature in a super clean bench, transferring into 5% bacteria solution, and culturing on a constant temperature shaking table at 37 ℃ and 140rpm for 3 days.
(4) And (4) preparing an alkaline liquid culture medium again, replacing 30% of the alkaline liquid culture medium into the degradation system in the step (3) after interface activation and sterilization, and continuously placing the alkaline liquid culture medium into a constant-temperature incubator at 37 ℃ and 140rpm for culture. This was repeated for 60 days.
Example 2
(1) Preparation of PET micro plastic: firstly, polyester superfine fibers and slices are refined, surface grease is removed, and the polyester superfine fibers and the slices are fully dried after being washed for multiple times; then, crushing the substrate to the size of micro plastic, collecting the screened PET powder, washing with water, and drying for later use.
(2) Interface activation: preparing 200ml of culture solution according to a culture formula of alkali-resistant Comamonas testosteroni in a 500ml conical flask, and adding 1% by volume of Tween 40; the system is placed in an ultrasonic instrument to ensure that the PET and the interfacial active agent are uniformly dispersed, and the system is placed on a constant temperature shaking table at 37 ℃ and 140rpm for treatment for 2 to 5 hours to ensure that the surface of the PET is fully emulsified. The formula of the culture medium is as follows: 1.00g/L of ammonium chloride, 1.4g/L of sodium hydroxide, 3.7g/L of potassium chloride, 0.50g/L of sodium chloride, 0.25g/L of magnesium sulfate heptahydrate, 1g/L of PET, 0.5 mu g/L of boric acid, 0.2 mu g/L of ferric chloride hexahydrate, 0.4 mu g/L of manganese sulfate pentahydrate, 0.4 mu g/L of zinc chloride, 40.0 mu g/L of copper sulfate pentahydrate and 0.2 mu g/L of ammonium molybdate.
(3) Biological-chemical combined degradation of PET (polyethylene terephthalate) micro-plastic: taking out the culture medium in the step (2), sterilizing at high temperature and high pressure for 20min, cooling to room temperature in a super clean bench, transferring into 5% bacteria solution, and culturing on a constant temperature shaking table at 37 ℃ and 140rpm for 3 days.
(4) And (4) preparing an alkaline liquid culture medium again, replacing 30% of the alkaline liquid culture medium into the degradation system in the step (3) after interface activation and sterilization, and continuously placing the alkaline liquid culture medium into a constant-temperature incubator at 37 ℃ and 140rpm for culture. This was repeated for 60 days.
Example 3
(1) Preparation of PET micro plastic: firstly, polyester superfine fibers and slices are refined, surface grease is removed, and the polyester superfine fibers and the slices are fully dried after being washed for multiple times; then, crushing the substrate to the size of micro plastic, collecting the screened PET powder, washing with water, and drying for later use.
(2) Interface activation: preparing 200ml of culture solution according to a culture formula of alkali-resistant Comamonas testosteroni in a 500ml conical flask, and adding 1% by volume of Tween 60; the system is placed in an ultrasonic instrument to ensure that the PET and the interfacial active agent are uniformly dispersed, and the system is placed on a constant temperature shaking table at 37 ℃ and 140rpm for treatment for 2 to 5 hours to ensure that the surface of the PET is fully emulsified. The formula of the culture medium is as follows: 1.00g/L of ammonium chloride, 1.4g/L of sodium hydroxide, 3.7g/L of potassium chloride, 0.50g/L of sodium chloride, 0.25g/L of magnesium sulfate heptahydrate, 1g/L of PET, 0.5 mu g/L of boric acid, 0.2 mu g/L of ferric chloride hexahydrate, 0.4 mu g/L of manganese sulfate pentahydrate, 0.4 mu g/L of zinc chloride, 40.0 mu g/L of copper sulfate pentahydrate and 0.2 mu g/L of ammonium molybdate.
(3) Biological-chemical combined degradation of PET (polyethylene terephthalate) micro-plastic: taking out the culture medium in the step (2), sterilizing at high temperature and high pressure for 20min, cooling to room temperature in a super clean bench, transferring into 5% bacteria solution, and culturing on a constant temperature shaking table at 37 ℃ and 140rpm for 3 days.
(4) And (4) preparing an alkaline liquid culture medium again, replacing 30% of the alkaline liquid culture medium into the degradation system in the step (3) after interface activation and sterilization, and continuously placing the alkaline liquid culture medium into a constant-temperature incubator at 37 ℃ and 140rpm for culture. This was repeated for 60 days.
Example 4
(1) Preparation of PET micro plastic: firstly, polyester superfine fibers and slices are refined, surface grease is removed, and the polyester superfine fibers and the slices are fully dried after being washed for multiple times; then, crushing the substrate to the size of micro plastic, collecting the screened PET powder, washing with water, and drying for later use.
(2) Interface activation: preparing 200ml of culture solution according to a culture formula of alkali-resistant Comamonas testosteroni in a 500ml conical flask, and adding 1% by volume of Tween 80; the system is placed in an ultrasonic instrument to ensure that the PET and the interfacial active agent are uniformly dispersed, and the system is placed on a constant temperature shaking table at 37 ℃ and 140rpm for treatment for 2 to 5 hours to ensure that the surface of the PET is fully emulsified. The formula of the culture medium is as follows: 1.00g/L of ammonium chloride, 1.4g/L of sodium hydroxide, 3.7g/L of potassium chloride, 0.50g/L of sodium chloride, 0.25g/L of magnesium sulfate heptahydrate, 1g/L of PET, 0.5 mu g/L of boric acid, 0.2 mu g/L of ferric chloride hexahydrate, 0.4 mu g/L of manganese sulfate pentahydrate, 0.4 mu g/L of zinc chloride, 40.0 mu g/L of copper sulfate pentahydrate and 0.2 mu g/L of ammonium molybdate.
(3) Biological-chemical combined degradation of PET (polyethylene terephthalate) micro-plastic: taking out the culture medium in the step (2), sterilizing at high temperature and high pressure for 20min, cooling to room temperature in a super clean bench, transferring into 5% bacteria solution, and culturing on a constant temperature shaking table at 37 ℃ and 140rpm for 3 days.
(4) And (4) preparing an alkaline liquid culture medium again, replacing 30% of the alkaline liquid culture medium into the degradation system in the step (3) after interface activation and sterilization, and continuously placing the alkaline liquid culture medium into a constant-temperature incubator at 37 ℃ and 140rpm for culture. This was repeated for 60 days.
The above is only a specific embodiment of the present invention, but the technical features of the present invention are not limited thereto. Any simple changes, equivalent substitutions or modifications made on the basis of the present invention to solve the same technical problems and achieve the same technical effects are all covered in the protection scope of the present invention.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116162580A (en) * | 2022-08-05 | 2023-05-26 | 南京工业大学 | Comamonas capable of degrading microplastic, construction method and application thereof |
| CN118272275A (en) * | 2024-05-23 | 2024-07-02 | 佛山科学技术学院 | PET-degrading microorganisms and their cultivation methods, uses, and PET-degrading methods |
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| CN105950473A (en) * | 2016-07-18 | 2016-09-21 | 齐齐哈尔大学 | Separation and screening method for dimethyl phthalate degradation microorganisms |
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| CN105950473A (en) * | 2016-07-18 | 2016-09-21 | 齐齐哈尔大学 | Separation and screening method for dimethyl phthalate degradation microorganisms |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116162580A (en) * | 2022-08-05 | 2023-05-26 | 南京工业大学 | Comamonas capable of degrading microplastic, construction method and application thereof |
| CN116162580B (en) * | 2022-08-05 | 2025-05-02 | 南京工业大学 | A strain of Comamonas capable of degrading microplastics and its construction method and application |
| CN118272275A (en) * | 2024-05-23 | 2024-07-02 | 佛山科学技术学院 | PET-degrading microorganisms and their cultivation methods, uses, and PET-degrading methods |
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