CN112745858A - Persistent organic pollutant contaminated site repairing agent and preparation method thereof - Google Patents
Persistent organic pollutant contaminated site repairing agent and preparation method thereof Download PDFInfo
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- CN112745858A CN112745858A CN202011478183.XA CN202011478183A CN112745858A CN 112745858 A CN112745858 A CN 112745858A CN 202011478183 A CN202011478183 A CN 202011478183A CN 112745858 A CN112745858 A CN 112745858A
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K17/00—Soil-conditioning materials or soil-stabilising materials
- C09K17/40—Soil-conditioning materials or soil-stabilising materials containing mixtures of inorganic and organic compounds
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/04—Enzymes or microbial cells immobilised on or in an organic carrier entrapped within the carrier, e.g. gel or hollow fibres
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
Abstract
The invention discloses a persistent organic pollutant polluted site repairing agent and a preparation method thereof, and relates to the technical field of environmental pollution treatment. The soil repairing tablet comprises a soil repairing tablet and comprises the components of a fixing agent, an adsorbent and a repairing strain. The soil restoration sheet with strong pertinence is prepared on the basis of the in-situ restoration technology by utilizing the degradation capability of microorganisms, is convenient to carry, and can effectively avoid loss in soil; the repairing strain is embedded and fixed by a fixing agent, adsorbed by an adsorbent and mutually matched with the sheet structure, so that the repairing strain has high-efficiency fixing and stabilizing capacity; by utilizing the genetic engineering technology and the molecular hybridization technology, corresponding genes of the existing bacteria, the water environment adjacent bacteria and the target soil bacteria in the polluted site are combined with each other to form the hybrid bacteria which can degrade organic pollutants in the polluted site and the water environment adjacent to the polluted site and restore the polluted site to the target environment, and the hybrid bacteria have high pertinence and pertinence.
Description
Technical Field
The invention belongs to the technical field of environmental pollution treatment, and particularly relates to a persistent organic pollutant contaminated site repairing agent and a preparation method thereof.
Background
Soil pollutants can be roughly divided into two categories, namely inorganic pollutants and organic pollutants, wherein the organic pollutants mainly comprise organic pesticides, phenols, cyanides, petroleum, synthetic detergents, 3, 4-benzopyrene, harmful microorganisms brought by municipal sewage, sludge and animal manure, and the like. When the soil contains excessive harmful substances and exceeds the self-cleaning capacity of the soil, the composition, the structure and the function of the soil are changed, the activity of microorganisms is inhibited, and the harmful substances or decomposition products thereof are gradually accumulated in the soil and are indirectly absorbed by the human body through the soil → plants → the human body or through the soil → water → the human body, so that the degree of harming the health of the human body is reached, namely the soil pollution.
In the prior art, a plurality of measures are provided for repairing polluted soil, which can be roughly divided into physical repair, chemical repair and biological repair, wherein the most common repair method which has the highest repair speed and is relatively healthy to the soil is a microbial repair technology in the biological repair; however, in the microbial repair technology, no matter in-situ repair or ex-situ repair is performed, compared with other repair methods, the repair agent is easy to run off along with water, so that long-term timeliness is difficult to guarantee, only temporary repair can be performed, and the repair capacity is relatively low; in view of the above, a persistent organic pollutant contaminated site repairing agent and a preparation method thereof are designed.
Disclosure of Invention
The invention aims to provide a persistent organic pollutant contaminated site repairing agent and a preparation method thereof, and solves the problems that the existing soil repairing agent is easy to run off, so that the repairing capability is influenced and the repairing cost is increased.
In order to solve the technical problems, the invention is realized by the following technical scheme:
the invention relates to a persistent organic pollutant contaminated site restoration agent, which comprises a soil restoration sheet, wherein the soil restoration sheet comprises a fixing agent, an adsorbent and a restoration strain, the restoration strain is wrapped in the fixing agent, and the fixing agent and the restoration strain are adsorbed in the adsorbent, namely the persistent organic pollutant contaminated site restoration agent is mainly constructed by wrapping and fixing the restoration strain by using the fixing agent, adsorbing a composite structure after wrapping and fixing by using the adsorbent and compacting the composite structure into the soil restoration sheet.
A preparation method of a persistent organic pollutant contaminated site repairing agent mainly comprises the following steps:
step one, selecting a target strain and preparing a repair strain;
step two, culturing the repairing strains, namely performing amplification culture on the repairing strains, so that the mass production is facilitated;
step three, the immobilization treatment of the repairing strains and the preparation of the soil repairing sheet can effectively avoid the phenomenon that the repairing agent is lost along with water during the actual use;
step four, detecting the ectopic microorganism repair test of the soil repair sheet, detecting the actual using effect of the soil repair sheet through the ectopic repair test, and simultaneously detecting the applicable environment of the soil repair sheet;
selecting sources of the target strains in the first step, wherein the selected sources comprise the existing flora in an organic pollutant polluted site, the target soil environment flora and the water environment flora close to the polluted site; the environment of the target soil is a remediation target environment of a polluted site, and the remediation strain has the capacity of degrading organic pollutants in a polluted area and an adjacent water environment on the basis of the target soil environment flora, so that the remediation capacity of the target soil is finally achieved;
the experimental environment of the ectopic microorganism remediation test in the fourth step is a contaminated site soil reactor, and related reaction variables can be controlled in a targeted manner to detect the remediation capability of the remediation agent.
Further, the first step mainly comprises the following steps:
s1, sampling from a polluted site, a target soil environment and a water environment close to the polluted site respectively;
s2, screening bacteria A, bacteria B and bacteria C with the strongest growth activity from the three environments in the step S1 by utilizing a microbial culture technology, wherein the bacteria A are the existing strains in the polluted site, the bacteria B are the strains in the target soil environment, and the bacteria C are the strains in the water environment close to the polluted site;
s3, extracting genes of A bacteria and C bacteria by using a genetic engineering technology and making a genetic information fragment, mainly extracting and shearing the genes of the A bacteria and the C bacteria for degrading organic pollutants into the genetic information fragment, and amplifying the genetic information fragment by using a PCR technology;
and S4, inserting the genetic information fragment prepared in the step S3 into the B bacteria and carrying out subculture to obtain repair bacteria, wherein the expression state and the result of related genes can be observed and detected through a gene probe so as to facilitate screening.
Further, the culturing of the repairing strains in the second step is the screening of the repairing strains in the step S4, and the culturing is mainly performed by controlling a plurality of groups of culturing environments and selecting the optimal strains from the culturing environments, wherein the culturing environments are mainly mixed components formed by different proportions of the polluted site soil, the target soil environment and the water source adjacent to the polluted site.
Further, the third step mainly includes the following steps:
step SS1, taking culture bacteria liquid of the optimal strains in the step III, adding a fixing agent into the culture bacteria liquid, and performing first-step immobilization treatment on the repaired strains by using an embedding method, wherein the embedding treatment can ensure the activity of the strains;
step SS2, taking the suspension formed after the first step of immobilization treatment in step SS1, adding an adsorbent into the suspension to adsorb and fix the embedded bacteria balls, and effectively avoiding the loss of the repair bacteria along with water during the actual work;
and SS3, filtering the suspension adsorbed and fixed in the SS2, taking bottom layer bacterial sludge, injecting the bottom layer bacterial sludge into a corresponding template, compacting and freeze-drying, wherein the template structure is manufactured according to the actual structure of the soil restoration sheet.
Furthermore, the fixing agent is sodium alginate, the adsorbent is diatomite, and the sodium alginate is used as the fixing agent mainly because the materials are convenient to obtain and low in cost, and can completely wrap the repairing strain; the main reason for using diatomite as the adsorbent is that the inner pores are large, and embedded bacteria balls can be adsorbed into the diatomite simultaneously.
Further, the control conditions of the ectopic microorganism repair test in the fourth step comprise water, temperature, oxygen content and pH value, different working environments are simulated by a controlled variable method, and the degradation capability of the ectopic microorganism repair test is enhanced by using subculture in the culture process.
Furthermore, the strain screening standards of the second step and the step S2 are both the growth and reproduction rate and the metabolic reaction rate of the microorganism.
The invention has the following beneficial effects:
the soil restoration sheet with strong pertinence is prepared on the basis of the in-situ restoration technology by utilizing the degradation capability of microorganisms, is convenient to carry, and can effectively avoid loss in soil; the repairing strain is embedded and fixed by a fixing agent, adsorbed by an adsorbent and mutually matched with the sheet structure, so that the repairing strain has high-efficiency fixing and stabilizing capacity;
on the other hand, the invention combines the corresponding genes of the existing bacteria, the bacteria near the water environment and the target soil bacteria in the polluted site with each other by utilizing the genetic engineering technology and the molecular hybridization technology to form the hybrid bacteria which can degrade the organic pollutants in the polluted site and the water environment near the polluted site and can restore the polluted site to the target environment, and has high pertinence and pertinence.
Of course, it is not necessary for any product in which the invention is practiced to achieve all of the above-described advantages at the same time.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the description of the present invention, it is to be understood that the terms "upper", "middle", "outer", "inner", and the like, indicate orientations or positional relationships, are used for convenience in describing the present invention and simplifying the description, but do not indicate or imply that the referenced components or elements must have a particular orientation, be constructed and operated in a particular orientation, and thus, should not be construed as limiting the present invention.
The invention relates to a persistent organic pollutant contaminated site restoration agent, which comprises a soil restoration sheet, wherein the soil restoration sheet comprises a fixing agent, an adsorbent and a restoration strain, the restoration strain is wrapped in the fixing agent, and the fixing agent and the restoration strain are adsorbed in the adsorbent, namely the persistent organic pollutant contaminated site restoration agent is mainly constructed by wrapping and fixing the restoration strain by using the fixing agent, adsorbing a wrapped and fixed composite structure by using the adsorbent and compacting the composite structure into the soil restoration sheet.
A preparation method of a persistent organic pollutant contaminated site repairing agent mainly comprises the following steps:
step one, selecting a target strain and preparing a repair strain;
step two, culturing the repairing strains, namely performing amplification culture on the repairing strains, so that the mass production is facilitated;
step three, the immobilization treatment of the repairing strains and the preparation of the soil repairing sheet can effectively avoid the phenomenon that the repairing agent is lost along with water during the actual use;
step four, detecting the ectopic microorganism repair test of the soil repair sheet, detecting the actual using effect of the soil repair sheet through the ectopic repair test, and simultaneously detecting the applicable environment of the soil repair sheet;
selecting sources of target strains in the first step, wherein the selected sources comprise the existing flora in an organic pollutant polluted site, the target soil environment flora and the water environment flora close to the polluted site; the environment of the target soil is a repairing target environment of a polluted site, and the target soil environment flora is mainly used as a basis, so that the repairing strain has the capability of degrading organic pollutants in a polluted area and a nearby water environment, and finally the repairing capability of the target soil is achieved;
the experimental environment of the ectopic microorganism remediation test in the fourth step is a soil reactor of a polluted site, and the relevant reaction variables can be controlled in a targeted manner to detect the remediation capability of the remediation agent.
Preferably, step one mainly comprises the following steps:
s1, sampling from a polluted site, a target soil environment and a water environment close to the polluted site respectively;
s2, screening bacteria A, bacteria B and bacteria C with the strongest growth activity from the three environments in the step S1 by utilizing a microbial culture technology, wherein the bacteria A are the existing strains in the polluted site, the bacteria B are the strains in the target soil environment, and the bacteria C are the strains in the water environment close to the polluted site;
s3, extracting genes of A bacteria and C bacteria by using a genetic engineering technology and making a genetic information fragment, mainly extracting and shearing the genes of the A bacteria and the C bacteria for degrading organic pollutants into the genetic information fragment, and amplifying the genetic information fragment by using a PCR technology;
and S4, inserting the genetic information fragment prepared in the step S3 into the B bacteria and carrying out subculture to obtain repair bacteria, wherein the expression state and the result of related genes can be observed and detected through a gene probe so as to facilitate screening.
Preferably, the culturing of the repairing strains in the second step is the screening of the repairing strains in the step S4, and the culturing environment is mainly a mixed component formed by different proportions of the polluted site soil, the target soil environment and the water source adjacent to the polluted site by controlling a plurality of groups of culturing environments to culture and selecting the optimal strains from the culturing environments.
Preferably, step three mainly comprises the following steps:
step SS1, taking culture bacteria liquid of the optimal strains in the step III, adding a fixing agent into the culture bacteria liquid, and performing first-step immobilization treatment on the repaired strains by using an embedding method, wherein the embedding treatment can ensure the activity of the strains;
step SS2, taking the suspension formed after the first step of immobilization treatment in step SS1, adding an adsorbent into the suspension to adsorb and fix the embedded bacteria balls, and effectively avoiding the loss of the repair bacteria along with water during the actual work;
and SS3, filtering the suspension adsorbed and fixed in the SS2, taking bottom layer bacterial sludge, injecting the bottom layer bacterial sludge into a corresponding template, compacting and freeze-drying, wherein the template structure is manufactured according to the actual structure of the soil restoration sheet.
Preferably, the fixing agent is sodium alginate, the adsorbent is diatomite, and the sodium alginate is used as the fixing agent mainly because of convenient material acquisition and low cost, and can completely wrap the repairing strain; the main reason for using diatomite as the adsorbent is that the inner pores are large, and embedded bacteria balls can be adsorbed into the diatomite simultaneously.
Preferably, the control conditions of the ectopic microorganism repair test in the fourth step comprise moisture, temperature, oxygen content and pH value, different working environments are simulated by a controlled variable method, and the degradation capability of the ectopic microorganism repair test is enhanced by subculturing in the culture process.
Preferably, the strain screening criteria of step two and step S2 are both the growth and reproduction rate and the metabolic reaction rate of the microorganism.
In the description herein, references to the description of "one embodiment," "an example," "a specific example" or the like are intended to mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
The preferred embodiments of the invention disclosed above are intended to be illustrative only. The preferred embodiments are not intended to be exhaustive or to limit the invention to the precise embodiments disclosed. Obviously, many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the invention and the practical application, to thereby enable others skilled in the art to best utilize the invention. The invention is limited only by the claims and their full scope and equivalents.
Claims (8)
1. The persistent organic pollutant contaminated site repairing agent comprises a soil repairing sheet and is characterized in that: the soil restoration sheet comprises a fixing agent, an adsorbent and a restoration strain, wherein the restoration strain is wrapped in the fixing agent, and the fixing agent and the restoration strain are adsorbed in the adsorbent.
2. A preparation method of a persistent organic pollutant contaminated site repairing agent is characterized by mainly comprising the following steps:
step one, selecting a target strain and preparing a repair strain;
step two, culturing the repairing strains;
step three, the immobilization treatment of the repairing strain and the preparation of a soil repairing sheet;
step four, detecting the ectopic microorganism repairing test of the soil repairing sheet;
selecting sources of the target strains in the first step, wherein the selected sources comprise the existing flora in an organic pollutant polluted site, the target soil environment flora and the water environment flora close to the polluted site; the environment of the target soil is a remediation target environment of a polluted site;
the experimental environment of the ectopic microorganism remediation test in the fourth step is a polluted site soil reactor.
3. The method for preparing the remediation agent for the site polluted by the persistent organic pollutant according to claim 2, wherein the step one mainly comprises the following steps:
s1, sampling from a polluted site, a target soil environment and a water environment close to the polluted site respectively;
s2, screening bacteria A, bacteria B and bacteria C with the strongest growth activity from the three environments in the step S1 by utilizing a microbial culture technology, wherein the bacteria A are the existing strains in the polluted site, the bacteria B are the strains in the target soil environment, and the bacteria C are the strains in the water environment close to the polluted site;
s3, extracting and making genetic information segments by using genes of bacteria A and C in the genetic engineering technology;
and S4, inserting the genetic information fragment prepared in the step S3 into the B bacteria and carrying out subculture to obtain the repairing bacteria.
4. The method for preparing the repairing agent for the contaminated site with persistent organic pollutants as claimed in claim 2, wherein the culturing of the repairing bacteria in the second step is the screening of the repairing bacteria in the step S4, and the optimal bacteria is selected from the culturing bacteria by controlling a plurality of groups of culturing environments.
5. The method for preparing the remediation agent for the site polluted by the persistent organic pollutant according to claim 2, wherein the third step mainly comprises the following steps:
step SS1, taking the culture bacterial liquid of the optimal strain in the step III, adding a fixing agent into the culture bacterial liquid, and carrying out first-step immobilization treatment on the repaired strain by using an embedding method;
step SS2, taking the suspension formed after the first step of immobilization treatment in the step SS1, and adding an adsorbent into the suspension to adsorb and immobilize the embedded bacteria balls;
and SS3, filtering the suspension adsorbed and fixed in the SS2, taking bottom layer bacterial sludge, injecting the bottom layer bacterial sludge into a corresponding template, compacting and freeze-drying.
6. The method for preparing the persistent organic pollutant contaminated site remediation agent according to claim 1, wherein the fixing agent is sodium alginate and the adsorbent is diatomaceous earth.
7. The method for preparing the remediation agent for the site polluted by the persistent organic pollutant according to claim 2, wherein the control conditions of the step four ectopic microorganism remediation test comprise moisture, temperature, oxygen content and pH value.
8. The method for preparing the remediation agent for the site polluted by the persistent organic pollutant according to claim 3, wherein the bacteria species screening criteria of the second step and the step S2 are both the growth and reproduction rate and the metabolic reaction rate of the microorganism.
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Cited By (1)
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Application publication date: 20210504 |