A kind of method for removing mercurous wastewater pollutants
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 method for removing mercurous 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 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 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 Sphingomonas, 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 Sphingomonas are Sphingomonas (Sphingomonas sp.) CGMCC N0.4589;
The bacillus pumilus is bacillus pumilus (Bacillus pumilus) ATCC27142;
The Phanerochaete chrysosporium is Phanerochaete chrysosporium (Phanerochaete chrysosporium)
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 Sphingomonas have been transferred to the sphingol of the encoding gene of following any one protein variant of coding
Sporangium (Sphingomonas sp.) CGMCC N0.4589, the protein variant is relative to PPKL1 albumen Original aminos
Sequence be mutated as follows respectively 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, albumen original amino acid referring to
GenBank:NP_744926.1.
Present invention simultaneously provides a kind of transgenic strain for being capable of efficient absorption mercury, the bacterial strain by by amino acid sequence such as
Genbank:Sequence shown in NP_744926.1 realizes its purpose in importeding into Sphingomonas.
And described amino acid sequence can have a mutation in following no site, 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.(21L/Q represents that the L amino acid substitutions in original series the 21st are Q amino acid).Amino acid sequence after these mutation
The bacterial strain that row are not willing the amino acid sequence after having imported the mutation equally has similar effect.And applicant is by substantial amounts of reality
Checking is real, and not all substitution all has similar effect, have Sphingomonas after substantial amounts of substitution and other
Bacterial strain together and without preferable sewage disposal effect.
Using DNAMAN software Design primers, BamHI and SalI restriction enzyme sites are separately added into, it is complete according to amino acid sequence
Nucleotide sequence PDA genes described in gene chemical synthesis, purpose fragment is obtained by carrying out amplification, and PCR amplifications obtain genes of interest
PDA (while corresponding mutational site is incorporated into gene order by multiplex PCR, so as to obtain different mutants base
Cause), double digestion PCR primer is carried out with BamHI and SalI, with the expression vector pBPSE for also passing through BamHI and SalI double digestions
It is connected, the recombinant plasmid transformed that will be proved to be successful is entered into the Sphingomonas, obtains the genetic engineering bacterium of degraded mercury.
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 Hg 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 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/L containing phosphor contaminant,
Hg contents are 20mg/ml, by solid-liquid separator, carry out separation of solid and liquid, remove 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 Hg contents are 15mg/mL, removal effect is not notable.
Embodiment 2
Using DNAMAN software Design primers, BamHI and SalI restriction enzyme sites are separately added into, it is complete according to amino acid sequence
Nucleotide sequence PDA genes described in gene chemical synthesis, purpose fragment is obtained by carrying out amplification, and PCR amplifications obtain genes of interest
PDA (while by multiplex PCR by corresponding mutational site 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 are incorporated into gene order
In, so as to obtain different mutants gene), carry out double digestion PCR primer with BamHI and SalI, with also pass through BamHI and
The cloning vector pBPSE of SalI double digestions is connected, and the recombinant plasmid transformed that will be proved to be successful enters into the sphingol list
In spore bacterium CGMCC N0.4589, the genetic engineering bacterium of degraded mercury is obtained.
The Sphingomonas 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 that embodiment 2 is prepared is respectively adopted is removed the reaction of mercury,
Be found through experiments that, 21L/Q, 59F/S, 75R/V, 121E/D, 165A/P, 199H/R, 242D/V, 421L/Y, 480A/S,
This several mutant bacteria of 544R/I, 602A/L, 690N/Q, 729T/G, 842A/G, 901Y/V are all bright with having relative to original bacteria
Show enhanced effect.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 one
Cause.
Each pollutant concentration of sewage:NH3-N is 300mg/L, sulfide 80mg/L, is 70mg/L, Hg containing phosphor contaminant
Content is 20mg/ml.
As can be seen from the above results, the Sphingomonas and other several bacterial strains composition for being built using genetic engineering
Microbial inoculum in addition to original preferable removal ammonia nitrogen, the effect of sulfide and phosphorus, also with the preferably effect of removal Hg
Really, and in removal ammonia nitrogen, sulfide and in terms of phosphorus between the Sphingomonas and other several bacterial strains of genetic engineering go back
With enhanced synergy.