CN106745817A - It is a kind of to remove the method containing cadmium wastewater pollutants - Google Patents
It is a kind of to remove the method containing cadmium wastewater pollutants Download PDFInfo
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- CN106745817A CN106745817A CN201710061963.6A CN201710061963A CN106745817A CN 106745817 A CN106745817 A CN 106745817A CN 201710061963 A CN201710061963 A CN 201710061963A CN 106745817 A CN106745817 A CN 106745817A
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- 238000000034 method Methods 0.000 title claims abstract description 21
- 229910052793 cadmium Inorganic materials 0.000 title claims abstract description 15
- BDOSMKKIYDKNTQ-UHFFFAOYSA-N cadmium atom Chemical compound [Cd] BDOSMKKIYDKNTQ-UHFFFAOYSA-N 0.000 title claims abstract description 15
- 239000003344 environmental pollutant Substances 0.000 title claims abstract description 10
- 231100000719 pollutant Toxicity 0.000 title claims abstract description 10
- 239000002351 wastewater Substances 0.000 title claims abstract description 7
- 239000007788 liquid Substances 0.000 claims abstract description 31
- 241000894006 Bacteria Species 0.000 claims abstract description 26
- 239000010865 sewage Substances 0.000 claims abstract description 25
- 239000007787 solid Substances 0.000 claims abstract description 12
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 8
- 239000002131 composite material Substances 0.000 claims abstract description 8
- 238000006243 chemical reaction Methods 0.000 claims abstract description 5
- 239000013618 particulate matter Substances 0.000 claims abstract description 4
- 238000004062 sedimentation Methods 0.000 claims abstract description 4
- 238000000926 separation method Methods 0.000 claims abstract description 4
- 238000009288 screen filtration Methods 0.000 claims abstract 2
- 241000222393 Phanerochaete chrysosporium Species 0.000 claims description 18
- 108090000623 proteins and genes Proteins 0.000 claims description 16
- 241000194103 Bacillus pumilus Species 0.000 claims description 6
- 241000589614 Pseudomonas stutzeri Species 0.000 claims description 6
- 241001509286 Thiobacillus denitrificans Species 0.000 claims description 6
- 150000001413 amino acids Chemical class 0.000 claims description 6
- 102000004169 proteins and genes Human genes 0.000 claims description 6
- 239000002994 raw material Substances 0.000 claims description 6
- 241000316848 Rhodococcus <scale insect> Species 0.000 claims description 5
- 230000035772 mutation Effects 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 3
- 239000002244 precipitate Substances 0.000 claims description 2
- 240000007594 Oryza sativa Species 0.000 claims 1
- 235000007164 Oryza sativa Nutrition 0.000 claims 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims 1
- 235000009566 rice Nutrition 0.000 claims 1
- CPJBEAMIONWDDK-UHFFFAOYSA-N [N].N.[P].[S] Chemical compound [N].N.[P].[S] CPJBEAMIONWDDK-UHFFFAOYSA-N 0.000 abstract description 2
- 238000001556 precipitation Methods 0.000 abstract 1
- 230000000694 effects Effects 0.000 description 9
- 239000000126 substance Substances 0.000 description 8
- 238000011282 treatment Methods 0.000 description 8
- 125000003275 alpha amino acid group Chemical group 0.000 description 7
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 6
- UCKMPCXJQFINFW-UHFFFAOYSA-N Sulphide Chemical compound [S-2] UCKMPCXJQFINFW-UHFFFAOYSA-N 0.000 description 6
- 230000001580 bacterial effect Effects 0.000 description 6
- 238000010353 genetic engineering Methods 0.000 description 6
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
- 230000029087 digestion Effects 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 239000002068 microbial inoculum Substances 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 235000017166 Bambusa arundinacea Nutrition 0.000 description 3
- 235000017491 Bambusa tulda Nutrition 0.000 description 3
- 241001330002 Bambuseae Species 0.000 description 3
- 235000015334 Phyllostachys viridis Nutrition 0.000 description 3
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 239000011425 bamboo Substances 0.000 description 3
- 239000003610 charcoal Substances 0.000 description 3
- 239000000356 contaminant Substances 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 229910001385 heavy metal Inorganic materials 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 230000000869 mutational effect Effects 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 229910052698 phosphorus Inorganic materials 0.000 description 3
- 239000011574 phosphorus Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000006467 substitution reaction Methods 0.000 description 3
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 239000003124 biologic agent Substances 0.000 description 2
- 238000010170 biological method Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
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- 238000007254 oxidation reaction Methods 0.000 description 2
- 238000000053 physical method Methods 0.000 description 2
- 239000013612 plasmid Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 108091008146 restriction endonucleases Proteins 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000004065 wastewater treatment Methods 0.000 description 2
- 241000195493 Cryptophyta Species 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 244000131316 Panax pseudoginseng Species 0.000 description 1
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 241000187693 Rhodococcus rhodochrous Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 241001135759 Sphingomonas sp. Species 0.000 description 1
- CVTZKFWZDBJAHE-UHFFFAOYSA-N [N].N Chemical class [N].N CVTZKFWZDBJAHE-UHFFFAOYSA-N 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
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- 238000007233 catalytic pyrolysis Methods 0.000 description 1
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- 230000003247 decreasing effect Effects 0.000 description 1
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- 238000004821 distillation Methods 0.000 description 1
- 238000000909 electrodialysis Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
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- 230000002262 irrigation Effects 0.000 description 1
- 238000003973 irrigation Methods 0.000 description 1
- 230000004630 mental health Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
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- 239000010802 sludge Substances 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 238000003911 water pollution Methods 0.000 description 1
Classifications
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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
- C02F9/00—Multistage treatment of water, waste water or sewage
-
- 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
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
-
- 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
- C07K14/21—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Pseudomonadaceae (F)
-
- 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
- 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
- C12N1/14—Fungi; Culture media therefor
-
- 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
- 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
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- 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
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/52—Treatment of water, waste water, or sewage by flocculation or precipitation of suspended impurities
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- 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
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F2001/007—Processes including a sedimentation step
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/101—Sulfur compounds
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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
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/105—Phosphorus compounds
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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
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/20—Heavy metals or heavy metal compounds
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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
- C02F2209/00—Controlling or monitoring parameters in water treatment
- C02F2209/06—Controlling or monitoring parameters in water treatment pH
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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
- C02F2209/00—Controlling or monitoring parameters in water treatment
- C02F2209/10—Solids, e.g. total solids [TS], total suspended solids [TSS] or volatile solids [VS]
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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
- C02F2301/00—General aspects of water treatment
- C02F2301/08—Multistage treatments, e.g. repetition of the same process step under different conditions
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2303/00—Specific treatment goals
- C02F2303/24—Separation of coarse particles, e.g. by using sieves or screens
Abstract
The method containing cadmium wastewater pollutants is removed the present invention relates to a kind of, is comprised the following steps:First by sewage by solid-liquid separator, separation of solid and liquid is carried out, remove bulk solid particulate matter, subsequent liquid enters sedimentation basin, precipitation 12 hours, then liquid is removed into solid floccules by circular hole screen pack, the liquid through circular hole filter screen filtration enters biological reaction pool, regulation PH is 7,10 grams of composite bacteria agent is added every time by every cubic metre of liquid, is added daily 1 time, continuously add one week, 3 days finally are stood, liquid is discharged.The method of the present invention can effectively remove the pollutant containing ammonia nitrogen sulphur phosphorus and cadmium, and input cost is low, possess preferable application prospect.
Description
Technical field
The invention belongs to environmental technology field, and in particular to a kind of method of removal dirt water pollutant.
Background technology
Along with industrial development and the improvement of people's living standards, trade effluent amount and city domestic sewage
Amount just surges with surprising rapidity, these sewage or or has polluted river for the survival of mankind, river and lake, structure
One of the reason for environment for human survival being assisted into prestige.
In order to meet continuous improvement of the public to environmental quality requirement, country has worked out increasingly stricter discharge mark to nitrogen
Standard, research and development are economical, efficiently denitrogenating treatment technology has turned into the emphasis and focus of Water Pollution Control Engineering area research.Though
So having many methods can effectively remove ammonia, and such as physical method has counter-infiltration, distillation, soil irrigation；Chemical method has ion to hand over
Change method, ammonia aeration, chemical precipitation method, breakpoint chlorination, electrodialysis, electrochemical treatments, catalytic pyrolysis；Biological method has nitrification and algae
Class is cultivated, but physical method treatment effect is not good, and than chemical method, biological method treatment waste water has the following advantages:1) it is every kind of
Chemical article is possible to failure when other chemical substances are run into both for the very strong product of property, and biological agent is to dirt
The removal for contaminating thing has light spectrality；2) chemical products can temporarily eliminate some harmful substances and cover stink, and lacking to hinder
The only generation of harmful substance；3) using after chemical products, residual is had in water body, secondary pollution may be caused.Biological agent institute
Containing natural microbial, without pathogenic bacteria and pathogen, these microorganisms under the catalytic action of enzyme, with the organic nutrition in sewage
Material is food, and after sewage is purified, these microbes are gradually decreased with the reduction of pollutant, until withering away；4)
Nontoxic, non-corrosiveness is easy to use, and equipment or engineering need not be added substantially, saves fund input.
Sewage mainly has sanitary sewage and industrial wastewater.Trade effluent comparison of ingredients is complicated, particularly substantial amounts of artificial conjunction
Enter environment into compound, this kind of material is mainly ammonia nitrogen class, sulfide and phosphorus-containing compound, because these materials are tied in itself
The complexity of structure, can not be decomposed by the microorganisms utilization in a short time, and traditional method of wastewater treatment is tamed and dociled with activated sludge culture
The microorganism of change effectively can not be removed to these pollutants, and these materials are accumulated in the environment for a long time, be relied to us
Very big pollution is caused with the ecological environment survived, very big harm is brought to the physical and mental health of the mankind.Quite a few industry of China
Contaminating enterprises would rather be punished also be unwilling to invest and administer waste water, even if there have sewage-treatment plant to run to be also abnormal.Therefore, open
Sending out a kind of, construction investment is few, operating cost is low, good treatment efficiency sewage disposal technology is extremely urgent.
The content of the invention
In order to overcome the deficiencies in the prior art, effectively simply remove ammonia nitrogen sulphur phosphorus and heavy metal in sewage etc. and pollute
Thing, the invention provides a kind of method for removing dirty water pollutant, its technical scheme is realized by the following method:
A kind of to remove the method containing cadmium wastewater pollutants, it comprises the following steps:
The preparation of composite bacteria agent:By mixed bacteria liquid and carrier according to 1:1 weight stirs than mixing, then stands 6
Hour, it is most rearmounted 4 DEG C at low temperature drying, dry after water content control 6%, obtain final product；The carrier by bamboo charcoal, shitosan and
Diatomite is according to 2:2:1 mass ratio is mixed to get, particle diameter preferably 10 mesh of bamboo charcoal；
Sewage treatment step:First by sewage by solid-liquid separator, separation of solid and liquid is carried out, remove bulk solid particulate matter
Matter, subsequent liquid enters sedimentation basin, precipitates 12 hours, then liquid is removed into solid floccules, circular hole mistake by circular hole screen pack
The Circularhole diameter of filter screen is 0.1mm, biological oxidation:Enter biological reaction pool through the liquid of circular hole screen pack, regulation pH is 7, is pressed
Every cubic metre of liquid adds 10 grams of composite bacteria agent every time, adds daily 1 time, continuously adds one week, finally 3 days is stood, by liquid
Discharge.
Preferably, the composite bacteria agent is mixed by the raw material bacterium of following weight portion:
10 parts of Rhodococcus sp, 9 parts of thiobacillus denitrificans, 7 parts of Pseudomonas stutzeri, 6 parts of Sphingol single-cell, bacillus pumilus
5 parts, 2 parts of Phanerochaete chrysosporium；The concentration of above-mentioned each raw material bacterium is controlled in (1-2) × 108Individual/ml.Above-mentioned strain can be with
It is the conventional bacterial strain of prior art；
Preferably,
The Rhodococcus sp is Rhodococcus sp (Rhodococcusrhodochrous) ATCC15906；
The thiobacillus denitrificans are thiobacillus denitrificans (Thiobacillus denitrificans) ATCC25259；
The Pseudomonas stutzeri is Pseudomonas stutzeri (Pseudomonas stutzeri) CCTCC NO:M209107；
The Sphingol single-cell is Sphingol single-cell (Sphingomonas sp.) CGMCC N0.4589；
The bacillus pumilus is bacillus pumilus (Bacillus pumilus) ATCC27142；
The Phanerochaete chrysosporium is Phanerochaete chrysosporium (Phanerochaetechrysosporium)
ATCC24725。
Bacterium of the present invention can be from the business such as CGMCC, CCTCC and American Type Culture collection warehousing (ATCC)
Approach is commercially available.
Preferably, described Phanerochaete chrysosporium is the encoding gene for being transferred to following any one protein variant of coding
Phanerochaete chrysosporium (Phanerochaete chrysosporium) bacterial strains of ATCC 24725, the protein variant is relative
In albumen original amino acid be mutated as follows respectively 71S/T, 89T/Y, 121T/A, 143D/S, 184S/Q, 239W/G,
246V/P, 269K/N, 304P/S, 353K/E, 369G/D, 372C/S, 422D/R, 448G/I, albumen original amino acid ginseng
See GenBank：NP_744955.1.
Present invention simultaneously provides a kind of transgenic strain for being capable of efficient absorption cadmium, the bacterial strain by by amino acid sequence such as
Genbank：Sequence shown in NP_744955.1 realizes its purpose in importeding into Phanerochaete chrysosporium.
And described amino acid sequence can have a mutation in following different sites, 71S/T, 89T/Y, 121T/A,
143D/S、184S/Q、239W/G、246V/P、269K/N、304P/S、353K/E、369G/D、372C/S、422D/R、448G/I。
(71S/T represents that the S amino acid substitutions in original series the 71st are T amino acid).Amino acid sequence after these mutation is not grudged
The bacterial strain of the amino acid sequence imported after the mutation equally has similar effect.And applicant is by substantial amounts of experiment card
Real, not all substitution all has similar effect, there is Phanerochaete chrysosporium and other bacterium after substantial amounts of substitution
Strain together and do not have preferable sewage disposal effect.
Using DNAMAN software Design primers, BamHI and SalI restriction enzyme sites are separately added into, according to amino acid sequence
The described nucleotide sequence HP genes of NP_744955.1 full genomes synthesis, purpose fragment, PCR amplifications are obtained by carrying out amplification
Acquisition genes of interest HP (while corresponding mutational site is incorporated into gene order by multiplex PCR, so as to obtain difference
Mutant gene), double digestion PCR primer is carried out with BamHI and SalI, with the expression for also passing through BamHI and SalI double digestions
Carrier PWB980 is connected, and the recombinant plasmid transformed that will be proved to be successful is entered into the Phanerochaete chrysosporium, obtains degraded cadmium
Genetic engineering bacterium.
The Amplification Culture of each strain of the invention is the cellar culture mode of this area, it is also possible to described in reference literature
Training method obtain.
What the present invention was obtained mainly has the beneficial effect that:The present invention processes sewage using pure microbial inoculum, has effectively purified dirt
Water, and dephosphorization, denitrogenation removal simultaneously, the heavy metal cadmium that can remove sewage Central China simultaneously on this basis solve traditional work
Dephosphorization and denitrification effect are difficult to while take into account the contradiction of heavy metals removal in skill, with preferable application prospect.
Specific embodiment
Embodiment 1
A kind of method for removing dirty water pollutant, it comprises the following steps:
The preparation of composite bacteria agent:By mixed bacteria liquid and carrier according to 1:1 weight stirs than mixing, then stands 6
Hour, it is most rearmounted 4 DEG C at low temperature drying, dry after water content control 6%, obtain final product；Carrier is by bamboo charcoal, shitosan and diatom
Soil is according to 2:2:1 mass ratio is mixed to get；Above-mentioned mixed bacteria liquid is mixed by the raw material bacterium of following weight portion:Rhodococcus sp
10 parts, 9 parts of thiobacillus denitrificans, 7 parts of Pseudomonas stutzeri, 6 parts of Sphingol single-cell, 5 parts of bacillus pumilus, yellow archespore hair is flat
2 parts of lead fungi；The concentration of above-mentioned each raw material bacterium is controlled 1 × 108Individual/ml.
Sewage disinfection treatment:First by sewage (NH3-N is 300mg/L, sulfide 80mg/L, is 70mg/L containing phosphor contaminant,
Cadmium content is 40mg/ml, by solid-liquid separator, carries out separation of solid and liquid, removes bulk solid particulate matter, and subsequent liquid enters
Sedimentation basin, is precipitated 12 hours, then liquid is removed into solid floccules by circular hole screen pack, and the Circularhole diameter of circular hole screen pack is
0.1mm；
Biological oxidation:Enter biological reaction pool through the liquid of circular hole screen pack, regulation pH is 7.0, by every cubic metre of liquid
10 grams of composite bacteria agent is added every time, is added I times daily, continuously added one week, finally stand 3 days, liquid is discharged.After testing,
The content of ammonia nitrogen in sewage is respectively 12.5mg/L, sulfide 4.5mg/L, is 2.5mg/L containing phosphor contaminant, and clearance reaches
More than 95%；And cadmium content is 37mg/mL, removal effect is not notable.
Embodiment 2
Using DNAMAN software Design primers, BamHI and SalI restriction enzyme sites are separately added into, according to amino acid sequence
The described nucleotide sequence HP genes of NP_744955.1 full genomes synthesis, purpose fragment, PCR amplifications are obtained by carrying out amplification
Genes of interest HP is obtained (while passing through multiplex PCR by corresponding mutational site 71S/T, 89T/Y, 121T/A, 143D/S, 184S/
Q, 239W/G, 246V/P, 269K/N, 304P/S, 353K/E, 369G/D, 372C/S, 422D/R, 448G/I are incorporated into gene sequence
In row, so as to obtain different mutants gene), double digestion PCR primer is carried out with BamHI and SalI, and also pass through BamHI
It is connected with the cloning vector PWB980 of SalI double digestions, the recombinant plasmid transformed that will be proved to be successful enters into the yellow archespore
In flat lead fungi (Phanerochaete chrysosporium) ATCC24725 of hair, the genetic engineering bacterium of degraded cadmium is obtained.
The Phanerochaete chrysosporium genetic engineering bacterium of embodiment 3 is verified with the wastewater treatment efficiency of remaining several microbial inoculum
According to the method for embodiment 1, corresponding sewage disposal experiment is carried out, its sewage belongs to same with the sewage of embodiment 1
One batch, the pollutant concentration with same concentrations.The each group of wherein microbial inoculum is grouped into identical with embodiment 1.
The genetic engineering bacterium that the different mutational site of the acquisition of embodiment 2 is respectively adopted is removed the reaction of cadmium, passes through
Experiment find, 71S/T, 89T/Y, 121T/A, 143D/S, 184S/Q, 239W/G, 246V/P, 269K/N, 304P/S, 353K/E,
369G/D, 372C/S, 422D/R, 448G/I this several mutant bacteria all have the effect for having relative to original bacteria and being remarkably reinforced.
It is not more than total processing time 4 days, its result is as follows：Sewage is with a collection of sewage, so that guaranteed conditions are consistent.
Each pollutant concentration of sewage：NH3-N is 300mg/L, sulfide 80mg/L, is 70mg/L, cadmium containing phosphor contaminant
Content is 40mg/ml.
As can be seen from the above results, the Phanerochaete chrysosporium for being built using genetic engineering is constituted with other several bacterial strains
Microbial inoculum in addition to original preferable removal ammonia nitrogen, the effect of sulfide and phosphorus, also with preferably removal cadmium
Effect, and in terms of removal ammonia nitrogen, sulfide and phosphorus the Phanerochaete chrysosporium of genetic engineering and other several bacterial strains it
Between also have enhanced synergy.
Claims (4)
1. a kind of to remove the method containing cadmium wastewater pollutants, it comprises the following steps:First by sewage by solid-liquid separator, enter
Row separation of solid and liquid, removes bulk solid particulate matter, and subsequent liquid enters sedimentation basin, precipitates 12 hours, then by liquid by circle
Hole screen pack removal solid floccules, biological reaction pool is entered through the liquid of circular hole filter screen filtration, and regulation PH is 7, by often standing
Square rice liquid adds 10 grams of composite bacteria agent every time, adds daily 1 time, continuously adds one week, finally stands 3 days, and liquid is discharged.
2. the method for claim 1, it is characterised in that：The composite bacteria agent is by the raw material bacterium mixing of following weight portion
Into:10 parts of Rhodococcus sp, 9 parts of thiobacillus denitrificans, 7 parts of Pseudomonas stutzeri, 6 parts of Sphingol single-cell, 5 parts of bacillus pumilus,
2 parts of Phanerochaete chrysosporium；The concentration of above-mentioned each raw material bacterium is controlled in (1-2) × 108Individual/ml.
3. method as claimed in claim 2, it is characterised in that：Described Phanerochaete chrysosporium is as follows to be transferred to coding
Phanerochaete chrysosporium (Phanerochaete chrysosporium) bacterium of ATCC 24725 of the encoding gene of protein variant
Strain, the protein variant is to be mutated as follows relative to albumen original amino acid:353K/E, albumen Original amino
Sequence is referring to GenBank：NP_744955.1.
4. a kind of protein variant, it is characterised in that：The protein variant be carried out relative to albumen original amino acid it is as follows
Mutation:353K/E, albumen original amino acid is referring to GenBank：NP_744955.1.
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| CN111066572B (en) * | 2019-12-27 | 2022-01-14 | 福州康来生物科技有限公司 | Cultivation process of agaricus blazei murill with low heavy metal content |
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| CN106746216B (en) | 2020-04-28 |
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| CN106630494A (en) | 2017-05-10 |
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