CN110496856A - Pesticide-polluted soil remediation method based on laccase degradation - Google Patents
Pesticide-polluted soil remediation method based on laccase degradation Download PDFInfo
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- CN110496856A CN110496856A CN201910672111.XA CN201910672111A CN110496856A CN 110496856 A CN110496856 A CN 110496856A CN 201910672111 A CN201910672111 A CN 201910672111A CN 110496856 A CN110496856 A CN 110496856A
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- 239000002689 soil Substances 0.000 title claims abstract description 109
- 108010029541 Laccase Proteins 0.000 title claims abstract description 70
- 238000006731 degradation reaction Methods 0.000 title claims abstract description 53
- 230000015556 catabolic process Effects 0.000 title claims abstract description 49
- 238000000034 method Methods 0.000 title claims abstract description 27
- 238000005067 remediation Methods 0.000 title claims description 18
- 239000000575 pesticide Substances 0.000 claims abstract description 36
- 230000000694 effects Effects 0.000 claims abstract description 17
- 230000008439 repair process Effects 0.000 claims abstract description 6
- RMOGWMIKYWRTKW-UONOGXRCSA-N (S,S)-paclobutrazol Chemical compound C([C@@H]([C@@H](O)C(C)(C)C)N1N=CN=C1)C1=CC=C(Cl)C=C1 RMOGWMIKYWRTKW-UONOGXRCSA-N 0.000 claims description 34
- 239000005985 Paclobutrazol Substances 0.000 claims description 34
- YNWVFADWVLCOPU-MAUPQMMJSA-N uniconazole P Chemical compound C1=NC=NN1/C([C@@H](O)C(C)(C)C)=C/C1=CC=C(Cl)C=C1 YNWVFADWVLCOPU-MAUPQMMJSA-N 0.000 claims description 33
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 30
- 108090000790 Enzymes Proteins 0.000 claims description 22
- 102000004190 Enzymes Human genes 0.000 claims description 22
- BRPQOXSCLDDYGP-UHFFFAOYSA-N calcium oxide Chemical compound [O-2].[Ca+2] BRPQOXSCLDDYGP-UHFFFAOYSA-N 0.000 claims description 10
- 239000000292 calcium oxide Substances 0.000 claims description 10
- ODINCKMPIJJUCX-UHFFFAOYSA-N calcium oxide Inorganic materials [Ca]=O ODINCKMPIJJUCX-UHFFFAOYSA-N 0.000 claims description 10
- 241000856147 Pleurotus sp. Species 0.000 claims description 6
- 241000233866 Fungi Species 0.000 claims description 3
- 239000003973 paint Substances 0.000 claims description 3
- 230000028327 secretion Effects 0.000 claims description 3
- 239000003814 drug Substances 0.000 claims description 2
- 239000008363 phosphate buffer Substances 0.000 claims description 2
- 229940079593 drug Drugs 0.000 claims 1
- 238000006243 chemical reaction Methods 0.000 abstract description 10
- 239000003344 environmental pollutant Substances 0.000 abstract description 8
- 231100000719 pollutant Toxicity 0.000 abstract description 8
- 238000006555 catalytic reaction Methods 0.000 abstract description 3
- 230000035484 reaction time Effects 0.000 abstract description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 2
- 238000009423 ventilation Methods 0.000 abstract 1
- 229910019142 PO4 Inorganic materials 0.000 description 8
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 8
- 229910052760 oxygen Inorganic materials 0.000 description 8
- 239000001301 oxygen Substances 0.000 description 8
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 8
- 239000010452 phosphate Substances 0.000 description 8
- 239000000872 buffer Substances 0.000 description 6
- 238000011109 contamination Methods 0.000 description 5
- 238000005286 illumination Methods 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 description 3
- 239000000356 contaminant Substances 0.000 description 3
- 230000000593 degrading effect Effects 0.000 description 3
- 230000007613 environmental effect Effects 0.000 description 3
- 230000002255 enzymatic effect Effects 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 239000002957 persistent organic pollutant Substances 0.000 description 3
- 239000000376 reactant Substances 0.000 description 3
- -1 small molecule compound Chemical class 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 150000003852 triazoles Chemical class 0.000 description 3
- FMMWHPNWAFZXNH-UHFFFAOYSA-N Benz[a]pyrene Chemical compound C1=C2C3=CC=CC=C3C=C(C=C3)C2=C2C3=CC=CC2=C1 FMMWHPNWAFZXNH-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 230000000996 additive effect Effects 0.000 description 2
- 230000003139 buffering effect Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 229910001385 heavy metal Inorganic materials 0.000 description 2
- 239000005556 hormone Substances 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 229910021645 metal ion Inorganic materials 0.000 description 2
- 125000005575 polycyclic aromatic hydrocarbon group Chemical group 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000003900 soil pollution Methods 0.000 description 2
- LINPIYWFGCPVIE-UHFFFAOYSA-N 2,4,6-trichlorophenol Chemical class OC1=C(Cl)C=C(Cl)C=C1Cl LINPIYWFGCPVIE-UHFFFAOYSA-N 0.000 description 1
- HFZWRUODUSTPEG-UHFFFAOYSA-N 2,4-dichlorophenol Chemical compound OC1=CC=C(Cl)C=C1Cl HFZWRUODUSTPEG-UHFFFAOYSA-N 0.000 description 1
- WXNZTHHGJRFXKQ-UHFFFAOYSA-N 4-chlorophenol Chemical compound OC1=CC=C(Cl)C=C1 WXNZTHHGJRFXKQ-UHFFFAOYSA-N 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- 108010031396 Catechol oxidase Proteins 0.000 description 1
- 102000030523 Catechol oxidase Human genes 0.000 description 1
- 229930191978 Gibberellin Natural products 0.000 description 1
- 208000012766 Growth delay Diseases 0.000 description 1
- 102000043368 Multicopper oxidase Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 239000004098 Tetracycline Substances 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 150000001336 alkenes Chemical class 0.000 description 1
- 150000004982 aromatic amines Chemical class 0.000 description 1
- MXWJVTOOROXGIU-UHFFFAOYSA-N atrazine Chemical compound CCNC1=NC(Cl)=NC(NC(C)C)=N1 MXWJVTOOROXGIU-UHFFFAOYSA-N 0.000 description 1
- 150000003851 azoles Chemical class 0.000 description 1
- 150000004054 benzoquinones Chemical class 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 230000006652 catabolic pathway Effects 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000012272 crop production Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- LHRCREOYAASXPZ-UHFFFAOYSA-N dibenz[a,h]anthracene Chemical compound C1=CC=C2C(C=C3C=CC=4C(C3=C3)=CC=CC=4)=C3C=CC2=C1 LHRCREOYAASXPZ-UHFFFAOYSA-N 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 229940124307 fluoroquinolone Drugs 0.000 description 1
- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 description 1
- 239000003448 gibberellin Substances 0.000 description 1
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 1
- 239000010842 industrial wastewater Substances 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 108700020788 multicopper oxidase Proteins 0.000 description 1
- XNGIFLGASWRNHJ-UHFFFAOYSA-N o-dicarboxybenzene Natural products OC(=O)C1=CC=CC=C1C(O)=O XNGIFLGASWRNHJ-UHFFFAOYSA-N 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000000447 pesticide residue Substances 0.000 description 1
- 150000002989 phenols Chemical class 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 239000005648 plant growth regulator Substances 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000000275 quality assurance Methods 0.000 description 1
- LISFMEBWQUVKPJ-UHFFFAOYSA-N quinolin-2-ol Chemical compound C1=CC=C2NC(=O)C=CC2=C1 LISFMEBWQUVKPJ-UHFFFAOYSA-N 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000002910 solid waste Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 229940124530 sulfonamide Drugs 0.000 description 1
- 150000003456 sulfonamides Chemical class 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 229960002180 tetracycline Drugs 0.000 description 1
- 229930101283 tetracycline Natural products 0.000 description 1
- 235000019364 tetracycline Nutrition 0.000 description 1
- 150000003522 tetracyclines Chemical class 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 239000002351 wastewater Substances 0.000 description 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
- B09C1/105—Reclamation of contaminated soil microbiologically, biologically or by using enzymes using fungi or plants
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Mycology (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Botany (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Soil Sciences (AREA)
- Environmental & Geological Engineering (AREA)
- Processing Of Solid Wastes (AREA)
Abstract
A method for remediating pesticide-contaminated soil based on laccase degradation, the method comprising: controlling the pH value of soil to be repaired to be 3.5-7.5, adding laccase and a mediator HBT (heterojunction bipolar transistor), adjusting the water content of the soil to be 15-60%, naturally illuminating and keeping continuous ventilation for 24-48 h in an environment of 10-40 ℃, and repairing the soil polluted by pesticide; the laccase is adopted to repair the soil polluted by the pesticide, so that the efficiency is high, the environment is friendly, and secondary pollution cannot be caused; the laccase is used, the soil property range is wide, the use effect in natural environment is good, and the catalytic reaction can be carried out under a mild condition; the HBT mediator is used to promote the binding site of laccase and target pollutants, so that the laccase reaction efficiency is maximized, and the reaction time of the whole reaction system is reduced; therefore, the method has the characteristics of short repair period, good repair effect and no secondary pollution.
Description
(1) technical field
The invention belongs to by the technical field of soil remediation of pollution by pesticides, and in particular to it is a kind of based on laccase degradation by agriculture
The soil remediation method of chemicals contamination.
(2) background technique
In recent years, the direct application of various pesticides and industrial wastewater, solid waste etc. enter agriculture by degradation pathways
Field causes agricultural land soil quality safety by serious challenge, has seriously threatened agricultural food product production safety.Research shows that farmland
Soil pollution is divided into organic contamination and inorganic pollution, organic contamination mainly include pesticide, organic compound (such as: NBFRs, PAHs,
PAEs, DPs etc.), inorganic pollution mainly includes heavy metal, chemical reagent (such as: acid, alkali, salt).Wherein, pollution by pesticides conduct
The typical event of Farmland Soil Pollution by more document reports, therefore is carried out to repair and just be shown about the soil of pollution by pesticides
It obtains especially urgent.
Triazole pesticide, which is widely applied to, kills pest and field of cutting weeds, much big in usable floor area in farmland
In pesticide to the contaminated area (in fact, pollution level of most of pollutant in farmland is far below place) in place.Paclobutrazol
Belong to the efficient triazole type plant growth regulator of broad spectrum activity with uniconazole P, is widely used in field-crop etc., both agricultures
Remaining half-life period is more than more than half a year (0.5~1.0 year) to medicine in the soil, similar to part POPs, be may cause serious
Soil environment ecological risk and crop production reduction, bring negative effect can not be ignored.Paclobutrazol and uniconazole P pesticide enter
Crop mainly is influenced in terms of endogenous hormones and enzymatic activity two into botanical system.The influence of endogenous hormones is mainly reflected in suppression
Gibberellin (GA processed3) biosynthesis, this will lead to plant growth delay or stagnate.The influence of enzymatic activity be mainly reflected in through
The change of triazole pesticide treated catalase activity and heteroauxin (IAA) oxidase active directly affects IAA
Content, this indirectly influences the growth regulating of plant.Therefore, paclobutrazol/uniconazole P residual in repairing polluted soil is reduced
To green agriculture is developed, improves crop growth quality assurance food safety and have great importance.
Contaminated soil can significantly change various organic pollutant concentration in soil using biological enzyme reparation, promote soil
Earth environmental quality.Have more documents and describes biological enzyme type, environmental factor, pollutant concentration to biological enzyme reparation Polluted Soil
The influence of earth.Macromolecule contaminant degradation can not only be small molecule compound by biological enzyme repairing polluted soil, and to environment
Side effect it is lower, more environmentally friendly relative to other recovery techniques, reduce to environment, there may be the Potential feasibilities of secondary pollution.
The research achievement for repairing polluted farmland soil for biological enzyme is rarely reported, and limits the further development of correlative study.
Laccase (Laccase) is a kind of polyphenol oxidase of cupric, belongs to blue blue multicopper oxidase family.Laccase has extensive
Substrate specificity, a variety of phenolic compounds and its derivative, aromatic amine and its derivative etc. can be catalyzed, make its oxidation generate
Corresponding benzoquinones and water.Therefore, as a kind of environmentally friendly catalyst, organic contamination of the laccase in processing waste water and soil
Object has a good application prospect.Laccase utilizes the active function of itself in substrate, and then various relevant organic contaminations of degrading
Object.Existing at present some literature reports the researchs about laccase degradation of contaminant, such as pesticide (DDTs, atrazine, chlopyrifos
Deng), polycyclic aromatic hydrocarbon (Polycyclic aromatic hydrocarbons, PAHs) (benzo [a] pyrene, dibenzo [a, h] anthracene
Deng), phthalic acid ester (Phthalate ester, PAEs) (DMP), antibiotic (sulfonamide, tetracycline,
Quinolone, and Fluoroquinolones) etc..However, limitation is equally existed in practical degradation process, it is main next
From in the limitation of pollutant and enzymatic activity.Due in environment pollutant it is many kinds of, the presence of polluting effect altogether results in enzyme
Transformation efficiency there are dual characters: positive effect and negative effect.Research shows that when laccase degrades to 2,4- Dichlorophenol, when other
In the presence of phenol, different phenol (4- chlorophenol, 2,4,6- trichlorophenols) has different influences to it.Laccase is active in degradation process
Variation decides type, transforming degree and the different products and compound composition ratio of compound product.Meanwhile degradation process
The suction-operated of middle pollutant or package action prevent laccase further to interact, this may be the weight for causing laccase to inactivate
Want reason.Soil property (pH, TOC, microbe species, content of heavy metal etc.) also can generate corresponding shadow to the activity of enzyme
It rings.Therefore, influence of the research environment condition to laccase degradable organic pollutant, and select more excellent condition, for development and perfect
Soil remediation has important reference significance.
(3) summary of the invention
In view of the deficiencies in the prior art, that the present invention provides a kind of repairing efficiencies is short, repairing effect is good, without secondary
The soil remediation method by pollution by pesticides based on laccase degradation of pollution.
Technical scheme is as follows:
A kind of soil remediation method by pollution by pesticides based on laccase degradation, the method are as follows:
The pH of soil to be repaired is controlled between 3.5~7.5, laccase, mediator HBT (I-hydroxybenzotriazole) is added, is adjusted
Whole soil moisture content is to 15~60%, in 10~40 DEG C of environment, natural lighting and keeps 24~48h of continuous ventilating, repairs quilt
The soil of pollution by pesticides；
The dosage of the laccase is calculated as 1~8U/g with the quality of soil to be repaired；
The dosage of the mediator HBT is calculated as 0.2~2.7mg/g with the quality of soil to be repaired；
The pH of the soil to be repaired first carries out coarse adjustment with calcium oxide or acetic acid, then is accurately adjusted with phosphate buffer
Section；
The laccase is the laccase of white-rot fungi Pleurotus sp. secretion, and enzyme activity is 3~4U/mL, the white-rot fungi
Pleurotus sp. belongs to the fungi of Basidiomycotina；
The pesticide is at least one of paclobutrazol, uniconazole P.
Compared with the existing technology, the invention has the following advantages that
(1), the present invention using laccase reparation by the soil of pollution by pesticides, can in 48h fast degradation pollutant, efficiency
It is high；Paint enzyme viability determines that the final product of paclobutrazol and uniconazole P pollutant can be H2O and CO2, environmental-friendly, Bu Huizao
At secondary pollution.
(2), the present invention uses laccase, and applicable soil property range is wide, and using effect is good in the natural environment, and catalysis is anti-
It should can be carried out compared under temperate condition.
(3), the present invention uses mediator, and different mediators can cause entire in the reaction process of Laccase Catalyzed degradation substrate
Catalysis reaction forward inversely reacts.The bound site of laccase and target contaminant is promoted in the present invention using HBT mediator
Point makes laccase reactions efficiency reach maximization, and reduces the reaction time of entire reaction system.
Therefore, the present invention has the characteristics that repairing efficiency is short, repairing effect is good and without secondary pollution.
(4) Detailed description of the invention
Fig. 1: degradation of the different laccase concentration to paclobutrazol and uniconazole P pesticide；
Fig. 2: influence of the temperature to paclobutrazol and uniconazole P in laccase degradation soil；
Fig. 3: the pH influence to paclobutrazol and uniconazole P in laccase degradation soil；
Fig. 4: influence of the humidity to paclobutrazol and uniconazole P in laccase degradation soil；
Fig. 5: influence of the illumination to paclobutrazol and uniconazole P in laccase degradation soil；
Fig. 6: influence of the continuous blowing air to paclobutrazol and uniconazole P in laccase degradation soil；
Fig. 7: Cd2+And Fe2+Influence to paclobutrazol and uniconazole P in laccase degradation soil；
Influence of Fig. 8: the ABTS or HBT to paclobutrazol and uniconazole P in laccase degradation soil.
(5) specific embodiment
The invention will be further described With reference to embodiment, not to the limitation of its protection scope.
In present embodiment:
The laccase is the laccase of white-rot fungi Pleurotus sp. secretion, and enzyme activity is 3~4U/mL；The white-rot fungi
Pleurotus sp. belongs to the fungi of Basidiomycotina.
The soil to be repaired is by the soil of two class pollution by pesticides of paclobutrazol and uniconazole P.
The dosage of the mediator HBT powder is calculated as 0.2~2.7mg/g with the quality of soil to be repaired.
It is repeated no more in embodiment.
Embodiment 1
The pH value of soil to be repaired is adjusted 4.5 or so with calcium oxide or acetic acid, then with phosphate concn is 100mM buffering
Liquid accurately adjusts pH=4.5, and the HBT of 0.2mg/g is added, and enzyme additive amount is respectively 0U/gSoil, 1U/gSoil, 2U/gSoil, 4U/gSoil, adjustment
Moisture content is placed in 25 DEG C of constant incubators and cultivates 48h to 15%, respectively 0h, 3h, 6h, 12h, for 24 hours, 48h this 6 when
Between point measurement paclobutrazol/uniconazole P concentration value.Experimental result as shown in Figure 1, paclobutrazol and uniconazole P pesticide through degrading for 24 hours
Afterwards, it substantially tends towards stability.After 48h occurs for entire degradation, the paclobutrazol degradation rate under different laccase concentration is respectively 10%,
59%, 36% and 66%, the degradation rate of uniconazole P is respectively 7.7%, 45%, 42% and 44%.It proves in above-mentioned each condition
Under, there is the effect for Pesticide Residue in Soil to be repaired of degrading within the scope of every g soil laccase concentration 1U~4U, wherein concentration is 4U/g soil
Laccase additive amount be more suitable for it is this based on laccase degradation the soil remediation method by pollution by pesticides.
Embodiment 2
The pH value of soil to be repaired is adjusted 4.5 or so with calcium oxide or acetic acid, then with phosphate concn is 100mM buffering
Liquid accurately adjusts pH=4.5, and addition laccase forms 8U/gSoilDegradation environment, and add the HBT of 0.2mg/g, keep contaminated soil
Moisture content be 15%, constant incubator is arranged different temperature conditions in batches: 10 DEG C, 25 DEG C and 40 DEG C, cultivating 48h.Respectively
Paclobutrazol/uniconazole P concentration value is measured (with embodiment 1) in different time points.Experimental result is as shown in Fig. 2, entire degradation hair
After raw 48h, paclobutrazol degradation rate under the conditions of 10 DEG C, 25 DEG C and 40 DEG C is respectively 20%, 66% and 23%, and uniconazole P is respectively
27%, 44% and 20%.It proves in 10 DEG C~40 DEG C environment, laccase still has certain effect by the soil of pollution by pesticides for repairing
Fruit, wherein 25 DEG C of environmental condition is more suitable for this soil remediation method by pollution by pesticides based on laccase degradation.
Embodiment 3
Soil to be repaired is sufficiently mixed and laccase is added and forms 8U/gSoilSystem, while add 0.2mg/g HBT be situated between
Body.It is again 100mM buffer to setting value, and with phosphate concn with the pH value of calcium oxide or acetic acid adjusting contaminated soil sample
Accurately adjusting to pH value is respectively 3.5,4.5,5.5,6.5 and 7.5 (selection standard pH measuring methods, with pH meter measured value).It keeps
Soil moisture content is 15%, and all samples are placed in 25 DEG C of constant incubators and cultivate 48h, in different time points (with real
Apply example 1) residual concentration of measurement paclobutrazol/uniconazole P in the soil, experimental result are as shown in Figure 3 respectively.Entire degradation occurs
After 48h, paclobutrazol degradation rate under the conditions of pH=3.5,4.5,5.5,6.5 and 7.5 is respectively 39%, 60%, 35%, 23% and
20%, uniconazole P degradation rate is respectively 33%, 46%, 38%, 27% and 24%.Prove the soil remediation method for pH value model
The soil to be repaired being trapped among between 3.5~7.5 has feasibility, and the soil to be repaired that pH value is 4.5 is more suitable for using this
The soil remediation method by pollution by pesticides based on laccase degradation.
Embodiment 4
Soil to be repaired and laccase are mixed to form 8U/gSoilSystem, and add the HBT of 0.2mg/g, while using calcium oxide
Or acetic acid adjusts the pH value of contaminated soil sample, then adding phosphate concn is that 100mM buffer accurately adjusts pH=4.5, is protected
Holding soil moisture content is respectively 15% and 60%, is placed in 25 DEG C of constant incubators in batches and cultivates 48h, and in different time points
Paclobutrazol/uniconazole P residual concentration (ngg in (with embodiment 1) measurement soil-1), experimental result is as shown in Figure 4.Entire drop
After 48h occurs for solution, paclobutrazol degradation rate under the conditions of humidity 15% and humidity 60% is respectively 66% and 6.5%, uniconazole P point
It Wei 44% and 15%.Prove humidity between 15%~60% by pesticide contaminated soil using laccase degradation technique into
Row is repaired, and is controlled the moisture content of soil to be repaired and be more suitable for this soil by pollution by pesticides based on laccase degradation for 15%
Earth restorative procedure.
Embodiment 5
Laccase is added in soil to be repaired forms 8U/gSoilReaction system, and add the HBT of 0.2mg/g.Using calcium oxide or
Acetic acid adjusts the pH value of contaminated soil sample, then with phosphate concn is that 100mM buffer accurately adjusts contaminated soil pH=
4.5, keeping soil moisture content is 15%.The temperature that incubator is arranged is 25 DEG C, and illumination and dark two kinds of environment items is respectively set
Part mode, wherein illumination condition is 16h illumination and 8 hours dark, and dark ambient conditions then pass through the shading treatment simulating black filaments of sun
Part culture 48h.Measure paclobutrazol/uniconazole P residual concentration in soil, result (with embodiment 1) in different time points respectively
As shown in Figure 5.After 48h occurs for entire degradation, paclobutrazol degradation rate under illumination and dark condition is respectively 66% and 34%, alkene
Imitating azoles is respectively 44% and 19%.Prove that normal lighting conditions are more suitable for this soil by pollution by pesticides based on laccase degradation
Earth restorative procedure.
Embodiment 6
Laccase is added in pedotheque to be repaired forms two groups of 8U/gSoilReaction system and two groups do not add the reactant of laccase
System adjusts the pH value of contaminated soil sample with calcium oxide or acetic acid, and is that 100mM buffer accurately adjusts dirt with phosphate concn
Soil pH=4.5 are contaminated, the HBT of 0.2mg/g is added.Soil moisture content is maintained at 15%, is respectively set that obstructed oxygen is enzyme, obstructed oxygen
Not enzyme, the continuously enzyme and continuous logical not enzyme four groups of experiments of oxygen of logical oxygen, cultivate 48h in 25 DEG C of incubators.Respectively in difference
Time point (with embodiment 1) measures paclobutrazol/uniconazole P residual concentration in soil, and result is as shown in Figure 6.Entire degradation hair
After raw 48h, paclobutrazol obstructed oxygen it is not enzyme, it is logical oxygen is not enzyme, the enzyme and logical oxygen of obstructed oxygen is enzyme under the conditions of degradation rate distinguish
It is 11%, 23%, 48% and 71%, uniconazole P is respectively 12%, 22%, 42% and 73%.Prove that ventilated environment is more suitable for
This soil remediation method by pollution by pesticides based on laccase degradation.
Embodiment 7
Laccase is added in pedotheque to be repaired forms two groups of 8U/gSoilReaction system and two groups do not add the reactant of laccase
System adjusts the pH value of contaminated soil sample with calcium oxide or acetic acid, and is that 100mM buffer accurately adjusts dirt with phosphate concn
Soil pH=4.5 are contaminated, the HBT of 0.2mg/g is added, keep soil moisture content 15%.Finally respectively addition Metal Ions Cd and
Fe, concentration are set as 0mg kg-1、1mg kg-1、5mg kg-1With 10mg kg-1.48h is cultivated in 25 DEG C of incubators, is existed respectively
Different time points (with embodiment 1) measure paclobutrazol/uniconazole P residual concentration in soil, and result is as shown in Figure 7.Entire drop
After 48h occurs for solution, paclobutrazol is in Cd2+Concentration is respectively 0mg kg-1、1mg kg-1、5mg kg-1With 10mg kg-1Condition decline
Solution rate is respectively 68%, 76%, 73% and 64%, and uniconazole P degradation rate is respectively 44%, 76%, 68% and 50%.In Fe2+It is dense
Degree is respectively 0mg kg-1、1mg kg-1、5mg kg-1With 10mg kg-1Under the conditions of, paclobutrazol degradation rate is respectively 68%, 79%,
81% and 63%, uniconazole P is respectively 44%, 67%, 55% and 55%.Proof has earth backing to be repaired existing for a small amount of metal ion
This soil remediation method by pollution by pesticides based on laccase degradation can be used in earth environment.
Embodiment 8
Laccase is added in pedotheque to be repaired forms two groups of 8U/gSoilReaction system and two groups do not add the reactant of laccase
System adjusts the pH value of contaminated soil sample with calcium oxide or acetic acid, and is that 100mM buffer accurately adjusts dirt with phosphate concn
Soil pH=4.5 are contaminated, ABTS mediator is added respectively and HBT mediator, mediator concentration is set as 0mg/g, 0.4mg/g, 2.7mg/g.
It keeps soil moisture content 15%, 48h is cultivated in 25 DEG C of incubators, measures soil (with embodiment 1) in different time points respectively
Paclobutrazol/uniconazole P residual concentration, result are as shown in Figure 8 in earth.After 48h occurs for entire degradation, paclobutrazol is dense in ABTS
Degree is respectively 0mg/g, and degradation rate is respectively 17%, 13% and 9.7% under the conditions of 0.4mg/g, 2.7mg/g, and uniconazole P is respectively
25%, 9.8% and 16%.It is respectively 0mg/g in HBT concentration, under the conditions of 0.4mg/g, 2.7mg/g, paclobutrazol degradation rate difference
It is 15%, 58% and 66%, uniconazole P is respectively 11%, 49% and 44%.It is this based on paint to prove that addition HBT mediator is suitable for
The soil remediation method by pollution by pesticides of enzyme degradation, and concentration 2.7mg/g is more excellent.
Claims (4)
1. a kind of soil remediation method by pollution by pesticides based on laccase degradation, which is characterized in that the method are as follows:
The pH of soil to be repaired is controlled between 3.5~7.5, addition laccase, mediator HBT, adjustment soil moisture content to 15~
60%, in 10~40 DEG C of environment, natural lighting simultaneously keeps 24~48h of continuous ventilating, repairs by the soil of pollution by pesticides；
The dosage of the laccase is calculated as 1~8U/g with the quality of soil to be repaired；
The dosage of the mediator HBT is calculated as 0.2~2.7mg/g with the quality of soil to be repaired.
2. as described in claim 1 based on laccase degradation the soil remediation method by pollution by pesticides, which is characterized in that it is described to
The pH of rehabilitating soil first carries out coarse adjustment with calcium oxide or acetic acid, then is accurately adjusted with phosphate buffer.
3. the soil remediation method by pollution by pesticides as described in claim 1 based on laccase degradation, which is characterized in that the paint
Enzyme is the laccase of white-rot fungi Pleurotus sp. secretion, and enzyme activity is 3~4U/mL, the white-rot fungi Pleurotus sp.
Belong to the fungi of Basidiomycotina.
4. the soil remediation method by pollution by pesticides as described in claim 1 based on laccase degradation, which is characterized in that the agriculture
Medicine is at least one of paclobutrazol, uniconazole P.
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| CN113025337A (en) * | 2021-02-04 | 2021-06-25 | 浙江卓锦环保科技股份有限公司 | Preparation method of soil remediation preparation and application of soil remediation preparation in soil remediation |
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| JP7503229B1 (en) | 2023-07-24 | 2024-06-20 | 南京▲農業▼大学 | Removal of polycyclic aromatic hydrocarbons from soil by a recyclable laccase system |
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| CN111202941A (en) * | 2019-12-27 | 2020-05-29 | 仲恺农业工程学院 | Method for degrading organophosphorus pesticide by using bacterial laccase |
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| CN112845566A (en) * | 2020-12-31 | 2021-05-28 | 浙江工业大学 | Method for repairing phthalate polluted soil by using lipase |
| CN113025337A (en) * | 2021-02-04 | 2021-06-25 | 浙江卓锦环保科技股份有限公司 | Preparation method of soil remediation preparation and application of soil remediation preparation in soil remediation |
| CN114226443A (en) * | 2021-12-22 | 2022-03-25 | 中电建十一局工程有限公司 | Inorganic and organic composite polluted soil restoration technology integration process |
| CN114871266A (en) * | 2022-04-29 | 2022-08-09 | 天津城建大学 | Method for repairing soil polluted by refractory organic matters by using immobilized laccase in cooperation with soil microbial electrochemical system |
| CN114871266B (en) * | 2022-04-29 | 2023-09-05 | 天津城建大学 | Method for repairing degradation-resistant organic matter polluted soil by immobilized laccase in cooperation with soil microbial electrochemical system |
| JP7503229B1 (en) | 2023-07-24 | 2024-06-20 | 南京▲農業▼大学 | Removal of polycyclic aromatic hydrocarbons from soil by a recyclable laccase system |
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