CN106745823B - Method for removing mercury-containing sewage pollutants - Google Patents
Method for removing mercury-containing sewage pollutants Download PDFInfo
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- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 title claims abstract description 9
- 229910052753 mercury Inorganic materials 0.000 title claims abstract description 9
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- 239000002994 raw material Substances 0.000 claims description 4
- 239000002131 composite material Substances 0.000 claims description 2
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- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 abstract description 10
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- 239000011574 phosphorus Substances 0.000 abstract description 10
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- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 abstract description 2
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Classifications
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F9/00—Multistage treatment of water, waste water or sewage
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
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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
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/001—Processes for the treatment of water whereby the filtration technique is of importance
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- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
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- C—CHEMISTRY; METALLURGY
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- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/66—Treatment of water, waste water, or sewage by neutralisation; pH adjustment
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- C—CHEMISTRY; METALLURGY
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- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
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- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
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- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/348—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the way or the form in which the microorganisms are added or dosed
Abstract
The invention relates to a method for removing mercury-containing sewage pollutants, which comprises the following steps of firstly carrying out solid-liquid separation on sewage through a solid-liquid separator to remove large solid particle substances, then enabling the liquid to enter a sedimentation tank for sedimentation for 12 hours, removing solid flocculates from the liquid through a circular hole filter screen, enabling the liquid filtered through the circular hole filter screen to enter a biological reaction tank, adjusting the pH to be 7, adding 10 g of a compound microbial inoculum per cubic meter of the liquid every time, adding the compound microbial inoculum for 1 time every day, continuously adding the compound microbial inoculum for one week, finally standing for 3 days, and discharging the liquid. The method can effectively remove pollutants containing ammonia nitrogen, sulfur and phosphorus and mercury, has low input cost and has better application prospect.
Description
Technical Field
The invention belongs to the technical field of environmental protection, and particularly relates to a method for removing pollutants in sewage.
Background
With the development of industrial production and the improvement of living standards of people, the amount of industrial sewage and the amount of urban sewage, which pollute rivers and lakes where people live or pollute the rivers, lakes, etc., have been or are becoming one of the reasons for harming the living environment of people, are rapidly increasing at an alarming speed.
In order to meet the continuously improved requirements of the public on the environmental quality, the nation sets up more and more strict emission standards for nitrogen, and research and development of economic and efficient nitrogen removal treatment technology becomes a key point and a hotspot of research in the field of water pollution control engineering. Although there are many methods for removing ammonia effectively, such as physical methods like reverse osmosis, distillation, soil irrigation; the chemical method comprises an ion exchange method, ammonia stripping, a chemical precipitation method, breakpoint chlorination, electrodialysis, electrochemical treatment and catalytic cracking; biological methods include nitrification and algae cultivation, but physical methods have poor treatment effect, compared with chemical methods, biological methods have the following advantages that 1) each chemical product is a product with strong pertinence, and can lose efficacy when meeting other chemical substances, and biological agents have spectral removal on pollutants; 2) the chemical product can temporarily eliminate certain harmful substances and cover odor, but cannot prevent the generation of the harmful substances; 3) after the chemical product is used, residues are left in the water body, which may cause secondary pollution. The biological preparation contains natural microorganisms without pathogenic bacteria and pathogens, the microorganisms take organic nutrients in the sewage as food under the catalysis of enzyme, and after the sewage is purified, the microorganisms are gradually reduced along with the reduction of the pollutants until the microorganisms are eliminated; 4) the paint is non-toxic, non-corrosive, convenient to use, basically does not need to add equipment or engineering, and saves the capital investment.
The sewage mainly comprises domestic sewage and industrial wastewater. The components of industrial sewage are relatively complex, particularly a large amount of artificially synthesized compounds enter the environment, the substances mainly comprise ammonia nitrogen, sulfide and phosphorus-containing compounds, and the substances cannot be decomposed and utilized by microorganisms in a short time due to the complexity of the structures of the substances, so that the microorganisms cultured and domesticated by the activated sludge in the traditional wastewater treatment method cannot effectively remove the pollutants, the substances are accumulated in the environment for a long time, great pollution is caused to the ecological environment which depends on the survival of people, and great harm is brought to the physical and mental health of human beings. A certain part of industrial pollution enterprises in China are penalized and are not willing to invest and treat wastewater, and even if a sewage treatment device runs abnormally. Therefore, the development of a sewage treatment technology with less construction investment, low operation cost and good treatment efficiency is urgent.
Disclosure of Invention
In order to overcome the defects of the prior art and effectively and simply remove pollutants such as ammonia nitrogen, sulfur, phosphorus, heavy metals and the like in the sewage, the invention provides a method for removing the pollutants in the sewage, which is realized by the following method:
a method for removing mercury-containing sewage pollutants, comprising the following steps:
the preparation of the compound microbial inoculum comprises the steps of mixing the mixed bacterial liquid and the carrier according to the weight ratio of 1:1, uniformly stirring, standing for 6 hours, and finally drying at low temperature of 4 ℃, wherein the water content is controlled at 6% after drying, so as to obtain the compound microbial inoculum; the carrier is prepared by mixing bamboo charcoal, chitosan and diatomite according to the mass ratio of 2:2:1, and the particle size of the bamboo charcoal is preferably 10 meshes;
the sewage pretreatment comprises the steps of firstly carrying out solid-liquid separation on sewage through a solid-liquid separator to remove large solid particle substances, then enabling the liquid to enter a sedimentation tank for sedimentation for 12 hours, then removing solid flocculate from the liquid through a circular hole filter screen, enabling the diameter of a circular hole of the circular hole filter screen to be 0.1mm, carrying out biological oxidation, enabling the liquid passing through the circular hole filter screen to enter a biological reaction tank, adjusting the pH to be 7, adding 10 g of composite microbial inoculum per cubic meter of the liquid every time, adding for 1 time every day, continuously adding for one week, finally standing for 3 days, and discharging the liquid.
Preferably, the compound microbial inoculum is prepared by mixing the following raw material bacteria in parts by weight:
10 parts of rhodococcus, 9 parts of thiobacillus denitrificans, 7 parts of pseudomonas stutzeri, 6 parts of sphingomonas, 5 parts of bacillus pumilus and 2 parts of phanerochaete chrysosporium; the concentration of each raw material bacteria is controlled at (1-2) × 108One per ml. The above mentioned species may be conventional strains of the prior art,
preferably, the first and second electrodes are formed of a metal,
the Rhodococcus is Rhodococcus rhodochrous (Rhodococcus rhodochrous) ATCC 15906;
the Thiobacillus denitrificans is Thiobacillus denitrificans (Thiobacillus Denitriclaims) ATCC 25259;
the Pseudomonas stutzeri is Pseudomonas stutzeri (Pseudomonas stutzeri) CCTCC NO: M209107;
the Sphingomonas is CGMCC N0.4589;
the Bacillus pumilus is Bacillus pumilus (Bacillus pumilus) ATCC 27142;
the Phanerochaete chrysosporium is Phanerochaete chrysosporium (Phanerochaete chrysosporium) ATCC 24725.
The bacteria of the invention can be purchased from commercial approaches such as CGMCC, CCTCC, American type culture collection and storage (ATCC) and the like.
Preferably, the Sphingomonas sp CGMCC N0.4589 is transformed with a gene encoding any one of the following protein variants, namely 21L/Q, 59F/S, 75R/V, 121E/D, 165A/P, 199H/R, 242D/V, 421L/Y, 480A/S, 544R/I, 602A/L, 690N/Q, 729T/G, 842A/G, 901Y/V, which are mutated with respect to the original amino acid sequence of the PPKL1 protein, wherein the original amino acid sequence of the protein is shown in GenBank: NP _ 744926.1.
The invention also provides a transgenic strain capable of efficiently absorbing mercury, and the strain is prepared by mixing amino acid sequences such as Genbank: the sequence shown in NP-744926.1 was introduced into Sphingomonas to achieve its object.
And the amino acid sequence may have mutations at positions 21L/Q, 59F/S, 75R/V, 121E/D, 165A/P, 199H/R, 242D/V, 421L/Y, 480A/S, 544R/I, 602A/L, 690N/Q, 729T/G, 842A/G, 901Y/V which are not used. (21L/Q indicates the substitution of the L amino acid to the Q amino acid at position 21 of the original sequence). These mutated amino acid sequences are truncated to give similar effects to strains into which the mutated amino acid sequences have been introduced. However, the applicant has confirmed through a large number of experiments that not all the substitutions have similar effects, and the sphingosine monospora having a large number of substitutions together with other strains do not have a good effect of sewage treatment.
Designing primers by using DNAMAN software, respectively adding BamHI enzyme cutting sites and SalI enzyme cutting sites, synthesizing the nucleotide sequence PDA gene according to the whole gene of an amino acid sequence, amplifying to obtain a target fragment, amplifying by PCR to obtain a target gene PDA (simultaneously introducing corresponding mutation sites into the gene sequence by multiple PCR to obtain different mutant genes), performing double enzyme cutting on PCR products by using the BamHI enzyme and the SalI enzyme, connecting the products with an expression vector pBPSE which is also subjected to double enzyme cutting by the BamHI enzyme and the SalI enzyme, and transforming a successfully verified recombinant plasmid into the sphingomonas to obtain the mercury-degrading gene engineering bacteria.
The scale-up culture of each strain of the present invention is a conventional culture method in the art, and can be obtained by a culture method described in the literature.
The method has the main beneficial effects that the sewage is treated by the pure microbial inoculum, the sewage is effectively purified, and the phosphorus and nitrogen are removed simultaneously, so that the heavy metal mercury in the sewage can be removed simultaneously, the contradiction that the phosphorus and nitrogen removal effects are difficult to simultaneously consider the heavy metal removal in the traditional process is solved, and the method has a better application prospect.
Detailed Description
Example 1
A method for removing contaminants from wastewater, comprising the steps of:
the preparation of the compound microbial inoculum comprises the steps of mixing the mixed bacterial liquid and the carrier according to the weight ratio of 1:1, uniformly stirring, standing for 6 hours, and finally drying at low temperature of 4 ℃, wherein the water content is controlled at 6% after drying, so as to obtain the compound microbial inoculum; the carrier is prepared by mixing bamboo charcoal, chitosan and diatomite according to the mass ratio of 2:2: 1; the mixed bacterial liquid is prepared by mixing 10 parts of rhodococcus, 9 parts of thiobacillus denitrificans, 7 parts of pseudomonas stutzeri, 6 parts of sphingomonas, 5 parts of bacillus pumilus and 2 parts of phanerochaete chrysosporium by weight; the concentration of each raw material bacteria is controlled at 1 × 108One per ml.
The sewage pretreatment comprises the steps of firstly carrying out solid-liquid separation on 300mg/L of sewage NH3-N, 80mg/L of sulfide, 70mg/L of phosphorus-containing pollutant and 20mg/ml of Hg by a solid-liquid separator to remove large solid particle substances, then feeding the liquid into a sedimentation tank for sedimentation for 12 hours, removing solid flocculate from the liquid by a round hole filter screen, wherein the diameter of a round hole of the round hole filter screen is 0.1mm,
and (3) biological oxidation, namely, allowing the liquid passing through the circular hole filter screen to enter a biological reaction tank, adjusting the pH to 7.0, adding 10 g of the compound microbial inoculum per cubic meter of the liquid every time, adding the compound microbial inoculum I times every day, continuously adding the compound microbial inoculum for one week, finally standing for 3 days, and discharging the liquid. Through detection, the ammonia nitrogen content in the sewage is respectively 12.5mg/L, the sulfide content is 4.5mg/L, the phosphorus-containing pollutant content is 2.5mg/L, and the removal rate is more than 95%; and the Hg content is 15mg/mL, and the removal effect is not obvious.
Example 2
Designing primers by using DNAMAN software, respectively adding BamHI enzyme cutting sites and SalI enzyme cutting sites, synthesizing the nucleotide sequence PDA gene according to the whole gene of an amino acid sequence, amplifying to obtain a target fragment, amplifying by PCR to obtain the target gene PDA (simultaneously introducing corresponding mutation sites 21L/Q, 59F/S, 75R/V, 121E/D, 165A/P, 199H/R, 242D/V, 421L/Y, 480A/S, 544R/I, 602A/L, 690N/Q, 729T/G, 842A/G and 901Y/V into the gene sequence by multiple PCR to obtain different mutant genes), carrying out double enzyme cutting on PCR products by using BamHI and SalI, connecting with a cloning expression vector pBPSE which is also subjected to double enzyme cutting by using BamHI and SalI, transforming a verified recombinant plasmid into the sphingomonas bacterium CGMCC N0.4589, obtaining the genetic engineering bacteria for degrading mercury.
Example 3 verification of Sewage treatment Effect of genetically engineered bacterium Sphingomonas and the rest of microbial Agents
According to the method of example 1, a corresponding sewage treatment experiment was performed, in which the sewage was the same batch as that of example 1 and had the same concentration of the contaminant. The compositions of the components of the microbial inoculum are completely the same as those of the example 1.
Experiments show that 21L/Q, 59F/S, 75R/V, 121E/D, 165A/P, 199H/R, 242D/V, 421L/Y, 480A/S, 544R/I, 602A/L, 690N/Q, 729T/G, 842A/G and 901Y/V mutant bacteria which are prepared in example 2 and have different mutation sites have obviously enhanced effects relative to original bacteria. The total treatment time was not more than 4 days, with the following results: the sewage is the same batch of sewage, thereby ensuring the consistent conditions.
The concentration of each pollutant in the sewage is as follows: NH3-N was 300mg/L, sulphide 80mg/L, phosphorus-containing contaminant 70mg/L, and Hg content 20 mg/ml.
From the results, the bacterial agent formed by the sphingosine monospora constructed by the genetic engineering and other strains has better Hg removing effect besides better original ammonia nitrogen, sulfide and phosphorus removing effect, and has enhanced synergistic effect between the sphingosine monospora constructed by the genetic engineering and other strains in the aspects of ammonia nitrogen, sulfide and phosphorus removing.
Claims (2)
1. A method for removing mercury-containing sewage pollutants is characterized by comprising the following steps: comprises the following steps of firstly passing the sewage through a solid-liquid separator,performing solid-liquid separation to remove large solid particle substances, then allowing the liquid to enter a sedimentation tank for sedimentation for 12 hours, removing solid flocculates from the liquid through a circular hole filter screen, allowing the liquid filtered by the circular hole filter screen to enter a biological reaction tank, adjusting the pH to 7, adding 10 g of the compound microbial inoculum per cubic meter of the liquid every time, adding the compound microbial inoculum for 1 time every day, continuously adding the compound microbial inoculum for one week, finally standing for 3 days, and discharging the liquid; the composite microbial inoculum is prepared by mixing 10 parts of rhodococcus, 9 parts of thiobacillus denitrificans, 7 parts of pseudomonas stutzeri, 6 parts of sphingomonas, 5 parts of bacillus pumilus and 2 parts of phanerochaete chrysosporium by weight; the concentration of each raw material bacteria is controlled at (1-2) × 108Per ml; the Sphingomonas is transferred into Sphingomonas (Sphingomonas sp.) CGMCC N0.4589 of a coding gene for coding a protein variant, wherein the protein variant is 199H/R by the following mutation relative to the original amino acid sequence of PPKL1 protein, and the original amino acid sequence of the protein is shown in GenBank: NP _ 744926.1.
2. A protein variant characterized in that: the protein variant is mutated by 199H/R relative to the original amino acid sequence of the protein, and the original amino acid sequence of the protein is shown in GenBank: NP _ 744926.1.
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