A kind of process the method for quinoline pollutant in sewage
Technical field
The invention belongs to sewage treatment area, be specifically related to the process of quinoline pollutant in sewage.
Background technology
Quinoline is a kind of colourless, has the hygroscopic liquid of penetrating odor.Dissolve in ethanol, ether, benzene, curing
Carbon, is slightly soluble in water.From the point of view of its chemical constitution, it belongs to heterocyclic arene type organic, by shape when coal, timber, PETROLEUM PROCESSING
Become.Quinoline in coking chemical waste water is then many to be produced during decoking oil wash benzene.The pollution of quinoline is mainly by air, water body, ground
The modes such as lower coal mining are propagated, and also contain the composition of quinoline in the gas of medicated cigarette discharge.Some research worker research finds liver
Dirty is the Target organ of quinoline, has a certain impact hepatocarcinoma.
Quinoline is typical multi-ring aromatic nitrogen-contg jeterocyclic chemistry compound, has that toxicity is big, teratogenecity and a spy such as carcinogenecity is strong
Point, it is difficult to biodegradation, can pass through soil pollution groundwater resources, have huge potential danger to human health and ecological environment
Evil, causes the concern of people the most day by day.Quinoline is the Typical Representative of nitrogen-containing heterocycle compound, and purposes is extremely extensive, is multiple doctor
Important source material in the production such as medicine, agricultural product, dyestuff and solvent.But quinoline and its derivates can carcinogenic, teratogenesis, mutagenesis,
And it is difficult to biodegradation, it is the severe pollutant in environment.Quinoline in environment is essentially from coal gas, Fossil fuel processing, coal
Tar residue and wood preservation facility.Additionally, quinoline and its derivates or coking chemical waste water, petroleum wastewater, pharmacy waste water etc.
Persistent organic pollutants composition in various industrial sewage.By scholar, Organic substance to 2 kinds of coking chemical waste waters forms and carries out respectively
Analyze, it is indicated that the content of quinolines occupies the second of organic pollution.Owing to quinoline contains I the strongest nitrogen of electronegativity
Atom makes its water solublity strengthen, and therefore this compounds is more easy to spread in the environment and persistently exists.Investigate discovery to exist
Quinoline and hydroxyquinoline thereof at concentrations up to several mg/L in subsoil water near creosote polluted place and soil.Except this it
Outward, quinoline and its derivates is also present in the tissue of urban air, tobacco smoke, sea water and fish, therefore finds and effectively removes
The method tool of quinoline is of great significance.
Now, there is the processing method of a lot of quinoline.It is roughly divided into Physical, chemical method and bioanalysis.Physical and change
Although method removal efficiency is high, but cost is high, easily produces secondary pollution.Biological treating have low cost, secondary pollution light,
The advantages such as Environmental compatibility is good, have become one of preferred option in pollutant abatement technology.Decision biochemical processing process success,
Effectively, be suitable for factor, in addition to process conditions and operation management, for the functional gene of the process such as contaminant degradation, conversion
Important function be also apparent from.Biological treatment because of its have that treating capacity is big, cost is relatively low, mild condition and not producing
The advantages such as secondary pollution and favored by people.In recent years, by specific merit on the basis of not changing existing treatment facility
The biological reinforcing technology of the interpolation of energy microorganism, it is possible to be greatly improved original biological treatment system and the removal of target contaminant is imitated
Really, sewage treatment capacity is greatly improved, thus is increasingly subject to people's attention.
Researcher begins to separate the microorganism of degraded quinoline from 20 century 70s both at home and abroad.Microorganism about quinoline
Degradation pathway and degradation kinetics, genetic research are also being goed deep into, but up to the present, the biodegradation of this compounds
Process study is the most fairly limited, and other functional studies for quinoline-degrading bacterium are the most less.
Summary of the invention
An object of the present invention is to provide one to utilize biotechnology to process the method for waste water in quinoline, to solve waste water
In the problem that is difficult to biodegradation and detoxification of the pollutant in quinoline.
The present invention adopts the following technical scheme that realization:
A kind of processing method containing quinoline compound waste water, comprises the steps:
(1) pretreatment: by filtering and precipitation, remove the float in handled sewage;
(2) main process: adding nitrogen oxide, be made into composite waste, be collected in intake pool, wherein, carbon-nitrogen ratio is 5-
10:1, the carbon in carbon-nitrogen ratio comes from quinoline compound, nitrogen comes from the nitrogen oxide of interpolation;
(3) composite waste is promoted in upflow type aerating biological filter by intake pump, oxygen supply aeration 6h;
(4) sewage after aeration proceeds to anoxic zone, adds the floating stuffing carrying denitrifying bacterium, processes 1 day;
(5) post processing: after sterile-processed for the sewage after the process of anoxic zone, directly discharge or reuse.
Preferably, described denitrifying bacterium is for having proceeded to coding any one PPKL1 protein variant following (relative to PPKL1
(its aminoacid sequence sees albumen original amino acid: GenBank:NP_742191.1) suddenly change 60E/ the most as follows
R, 69F/P, 82P/G, 120I/F, 147L/A, 178R/S, 189E/V, 229I/Q, 243V/Q, 251S/E, 259E/P) (60E/R
Representing and replace with R aminoacid at the E aminoacid of original series the 60th, other implication is similar) the CCTCC M of encoding gene
2010209 bacterial strains.
The present invention additionally provides a kind of quinoline-degrading albumen PPKL1, its aminoacid sequence such as NP_742191.1.This albumen
It is that the differential protein expression screening induced by quinoline from pseudomonas putida is obtained.This albumen has stronger removal and divides
Solve the ability of quinoline.
The invention provides a series of PPKL1 protein variant (to enter respectively relative to PPKL1 albumen original amino acid
Capable suddenly change as follows 60E/R, 69F/P, 82P/G, 120I/F, 147L/A, 178R/S, 189E/V, 229I/Q, 243V/Q, 251S/E,
259E/P), it has the higher ability removing decomposition quinoline.By the coded sequence of shown variant amino acids at CCTCC M
When 2010209 bacterial strains are expressed, find that recombinant bacterium has relative to the polyP bacteria of unconverted described gene and preferably remove decomposition
The ability of quinoline, has more preferable effect.
Importing in denitrifying bacterium by this albumen, this bacterial strain not only has the effect preferably going to denitrogenate phosphorus, simultaneously can also
Enough effective quinoline removed in sewage, have preferable application prospect.
Said method has the advantages that
1, by the processing method of the present invention, substantially increase the degradation rate of quinoline, reduce the response time.
2, the albumen of a kind of quinoline of degrading it is provided and converts bacterial strain, there is higher quinoline removal efficiency and nitrogen phosphorus ligands
Effect.
By specific embodiment, the present invention will be described in detail in order to make it easy to understand, following.Needs refer in particular to
Going out, instantiation, merely to explanation, is not intended that limitation of the scope of the invention.Obviously the ordinary skill people of this area
The present invention can be made various correction and change according to illustrating herein by member within the scope of the invention, and these are revised
Also include in the scope of the present invention with changing.
Detailed description of the invention
The process of embodiment 1 quinoline compound waste water
(1) pretreatment: taking containing quinoline concentration is that 100mg/L sewage is filtered, by often by conventional filtering ponds
The sedimentation tank of rule precipitates, and removes the float in handled sewage;
(2) main process: the sewage after the process of float will be eliminated, add nitrogen oxide (if nitrogen content is enough,
Can also add), be made into composite waste, be collected in intake pool, wherein, carbon-nitrogen ratio is 8:1, the carbon in carbon-nitrogen ratio from
The nitrogen oxide of interpolation is come from quinoline compound, nitrogen;
(3) composite waste is promoted in upflow type aerating biological filter by intake pump, oxygen supply aeration 6h;
(4) sewage after aeration proceeds to anoxic zone, adds the floating stuffing carrying denitrifying bacterium, processes 1 day;Described instead
Nitrifier is the bacterial strain of culture presevation numbered CCTCC M 2010209, specifically may refer to CN102115719A;
(5) post processing: after sterile-processed for the sewage after the process of anoxic zone, by measuring, wherein the concentration of quinoline is
0.5mg/L。
The acquisition of embodiment 2 quinoline-degrading gene PPKL1
Take pseudomonas putida, use quinoline as derivant, carry out inducing culture, respectively with induction and the bacterium do not induced
Strain as a comparison, carries out the extraction of total protein, carries out bidirectional two-dimensional electrophoresis, by comparison, it was found that differentially expressed protein at 10.
Wherein choose the point that wherein differential expression is bigger to check order.Having obtained corresponding aminoacid sequence, its sequence is such as
Shown in Genbank:NP_742191.1.This gene carries out the prokaryotic expression of routine, and concrete steps are all conventional technique.Logical
Crossing determination of activity to find, this albumen has stronger quinoline removal effect.
The activity raising of embodiment 3PPKL1 protein mutation
By multiplex PCR by corresponding 60E/R, 69F/P, 82P/G, 120I/F, 147L/A, 178R/S, 189E/V,
229I/Q, 243V/Q, 251S/E, 259E/P, 121E/S, mutational site is introduced between in albumen shown in NP_742191.1, from
And construct different mutant proteins (preparation method referring to prior art can obtain).Prepare and obtain different prominent
Variant gene, is then connected respectively at carrier, and the recombinant plasmid transformed being proved to be successful is entered denitrifying bacterium CCTCC M
In 2010209, it is thus achieved that the genetic engineering bacterium of degraded quinoline.
The checking of embodiment 4 quinoline-degrading effect
Multiple engineering bacterias embodiment 3 obtained are respectively according to the method for embodiment 1, respectively under different quinoline concentration
The effect of the corresponding quinoline of degrading of checking.Using the Ep2 bacterium of CN 102206657A as comparison.Concrete outcome is as shown in the table:
From above result it can be seen that the denitrifying bacterium major part having imported the albumen PPKL1 of sudden change all has enhancing
Remove quinoline effect, this absolutely proves not to be that arbitrary change all has preferable effect.