CN105060620B - It is a kind of to remove the method containing arsenic wastewater pollutants - Google Patents
It is a kind of to remove the method containing arsenic wastewater pollutants Download PDFInfo
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Abstract
The method containing arsenic 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 arsenic, 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 arsenic 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, most rearmounted 4 °C of lower low temperature dryings, water content control is obtained final product 6% after drying；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 disinfection treatment: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 I times daily, 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 Sphingomonas, bacillus pumilus
5 parts, 2 parts of Phanerochaete chrysosporium；The concentration of above-mentioned each raw material bacterium is controlled in (1-2) X 108Individual/ml.Above-mentioned strain can
To be 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 Sphingomonas are Sphingomonas (Sphingomonas sp.) CGMCC N0.4589；
The bacillus pumilus is bacillus pumilus (Bacillus pumilus) ATCC27142；
The yellow flat lead fungi of raw wool of embracing embraces the flat lead fungi of raw wool (Phanerochaete chrysosporium) for yellow
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, the Rhodococcus sp is the ATCC15906 of the encoding gene for being transferred to following any one protein variant of coding
Bacterial strain, the protein variant be mutated as follows respectively relative to albumen original amino acid 62K/G, 80E/V, 93V/H,
120R/I, 131F/P, 143Q/S, 152P/Q, 169G/I, 179Q/A, 192Q/G, 241W/S, 258R/T, 263R/P, proteinogen
Beginning, amino acid sequence was referring to GenBank：NP_745067.1.
Present invention simultaneously provides a kind of transgenic strain for being capable of efficient absorption arsenic, the bacterial strain by by amino acid sequence such as
Genbank：Sequence shown in NP_745067.1 realizes its purpose in importeding into Rhodococcus sp.
And described amino acid sequence can have a mutation in following no site, 62K/G, 80E/V, 93V/H,
120R/I、131F/P、143Q/S、152P/Q、169G/I、179Q/A、192Q/G、241W/S、258R/T、263R/P.（62K/G
The K amino acid substitutions represented in original series the 62nd are G amino acid）.Amino acid sequence after these mutation does not grudge importing
The bacterial strain of the amino acid sequence after the mutation equally has similar effect.And applicant is by substantial amounts of it is experimentally confirmed that not
Be it is all of substitution all have similar effect, have the Rhodococcus sp after substantial amounts of substitution together with other bacterial strains and without compared with
The effect of good sewage disposal.
Using DNAMAN software Design primers, restriction enzyme site is separately added into, it is complete according to amino acid sequence NP_745067.1
Nucleotide sequence PHY genes described in gene chemical synthesis, purpose fragment is obtained by carrying out amplification, and PCR amplifications obtain genes of interest
PHY（Corresponding mutational site is incorporated into gene order by multiplex PCR simultaneously, so as to obtain different mutants base
Cause）, double digestion PCR primer is carried out with restriction endonuclease, with the expression vector for also passing through double digestionpEASYIt is connected, will be proved to be successful
Recombinant plasmid transformed enter into the Rhodococcus sp, obtain degraded arsenic 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 arsenic 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, most rearmounted 4 °C of lower low temperature dryings, water content control is obtained final product 6% after drying；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 Sphingomonas, 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:It is 300mg/L, sulfide 80mg/L first by sewage NH3-N, is 70mg containing phosphor contaminant
/ L, arsenic content is 20mg/ml, by solid-liquid separator, carries out separation of solid and liquid, removes bulk solid particulate matter, subsequent liquid
Into sedimentation basin, precipitate 12 hours, then liquid is removed into solid floccules by circular hole screen pack, the circular hole of circular hole screen pack is straight
Footpath 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 daily 1 time, 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, clearance containing phosphor contaminant
Reach more than 96%；And arsenic content is 17mg/mL, removal effect is not notable.
Embodiment 2
Using DNAMAN software Design primers, restriction enzyme site is separately added into, it is complete according to amino acid sequence NP_745067.1
Nucleotide sequence PHY genes described in gene chemical synthesis, purpose fragment is obtained by carrying out amplification, and PCR amplifications obtain genes of interest
PHY（Simultaneously by multiplex PCR by corresponding mutational site 62K/G, 80E/V, 93V/H, 120R/I, 131F/P, 143Q/S,
152P/Q, 169G/I, 179Q/A, 192Q/G, 241W/S, 258R/T, 263R/P are incorporated into gene order, so as to obtain not
Same mutant gene）, double digestion PCR primer is carried out with the restriction enzyme site for adding, carried with the clonal expression for also passing through double digestion
BodypEASYIt is connected, the recombinant plasmid transformed that will be proved to be successful is entered into the Rhodococcus sp ATCC15906 bacterial strains, is degraded
The genetic engineering bacterium of arsenic.
The Rhodococcus genes engineering bacteria 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 described in embodiment 2 is respectively adopted is removed the reaction of arsenic, passes through
Experiment find, 62K/G, 80E/V, 93V/H, 120R/I, 131F/P, 143Q/S, 152P/Q, 169G/I, 179Q/A, 192Q/G,
241W/S, 258R/T, 263R/P this several mutant bacteria all have the effect for having relative to original bacteria and being remarkably reinforced.During total treatment
Between be not more than 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 be 300mg/L, sulfide 80mg/L, containing phosphor contaminant be 70mg/
L, arsenic content is 20mg/ml.
| Strain name | NH3-N residual concentrations（mg/L） | Sulfide residual concentration（mg/L） | Residual concentration containing phosphor contaminant（mg/L） | Arsenic residual concentration（mg/L） |
| Rhodococcus sp original strain | 12.5 | 4.5 | 2.5 | 17 |
| HQJ-4（62K/G） | 0.9 | 0.7 | 0.5 | 0.7 |
| HQJ-8（80E/V） | 0.8 | 0.7 | 0.4 | 0.4 |
| HQJ-9（93V/H） | 0.7 | 0.8 | 0.5 | 0.6 |
| HQJ-11（120R/I） | 0.6 | 0.6 | 0.4 | 0.5 |
| HQJ-15（131F/P） | 0.8 | 0.8 | 0.6 | 0.8 |
| HQJ-17（143Q/S） | 0.9 | 0.7 | 0.5 | 0.9 |
| HQJ-22（152P/Q） | 0.8 | 0.6 | 0.6 | 1.0 |
| HQJ-23（169G/I） | 0.8 | 0.6 | 0.5 | 0.9 |
| HQJ-28（179Q/A） | 0.8 | 0.5 | 0.5 | 0.7 |
| HQJ-31（192Q/G） | 0.7 | 0.6 | 0.4 | 0.8 |
| HQJ-40（241W/S） | 0.9 | 0.7 | 0.6 | 0.7 |
| HQJ-45（258R/T） | 0.8 | 0.8 | 0.5 | 0.6 |
| HQJ-47（263R/P） | 0.8 | 0.8 | 0.4 | 0.5 |
As can be seen from the above results, the microbial inoculum that the Rhodococcus sp for being built using genetic engineering is constituted with other several bacterial strains is removed
Outside effect with original preferable removal ammonia nitrogen, sulfide and phosphorus, also with the effect for preferably going arsenic removal, and
Also there is enhanced collaboration in removal ammonia nitrogen, sulfide and in terms of phosphorus between the Rhodococcus sp and other several bacterial strains of genetic engineering
Effect.
Claims (2)
1. a kind of to remove the method containing arsenic 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 I times daily, continuously adds one week, finally stands 3 days, and liquid is discharged,
Described composite bacteria agent is mixed by the raw material bacterium of following weight portion:10 parts of Rhodococcus sp, 9 parts of thiobacillus denitrificans, Amur is false single
7 parts of born of the same parents bacterium, 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
Control in (1-2) × 108Individual/ml, the Rhodococcus sp is the encoding gene for being transferred to the following protein variant of coding
ATCC15906 bacterial strains, the protein variant is to be mutated as follows relative to albumen original amino acid:62K/G, proteinogen
Beginning, amino acid sequence was referring to GenBank：NP_745067.1.
2. 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:62K/G, albumen original amino acid is referring to GenBank：NP_745067.1.
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| CN201510441623.7A CN105060620B (en) | 2015-07-26 | 2015-07-26 | It is a kind of to remove the method containing arsenic wastewater pollutants |
| CN201611187827.3A CN106587506B (en) | 2015-07-26 | 2015-07-26 | A method of removal wastewater pollutants containing arsenic |
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