CN106145523B - A kind of method of quinoline pollutant in processing sewage - Google Patents

A kind of method of quinoline pollutant in processing sewage Download PDF

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CN106145523B
CN106145523B CN201610600468.3A CN201610600468A CN106145523B CN 106145523 B CN106145523 B CN 106145523B CN 201610600468 A CN201610600468 A CN 201610600468A CN 106145523 B CN106145523 B CN 106145523B
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ppkl1
quinoline
sewage
amino acid
protein variant
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CN106145523A (en
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窦欣童
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Shenzhen Hemin Biotechnology Co., Ltd.
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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
    • C02F3/30—Aerobic and anaerobic processes
    • C02F3/302—Nitrification and denitrification treatment
    • C02F3/305—Nitrification and denitrification treatment characterised by the denitrification
    • 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
    • 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
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C12N9/1229—Phosphotransferases with a phosphate group as acceptor (2.7.4)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y207/00—Transferases transferring phosphorus-containing groups (2.7)
    • C12Y207/04—Phosphotransferases with a phosphate group as acceptor (2.7.4)
    • C12Y207/04001—Polyphosphate kinase (2.7.4.1)
    • 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
    • C02F1/72—Treatment of water, waste water, or sewage by oxidation
    • 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
    • C02F2101/105—Phosphorus compounds
    • 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
    • C02F2101/30—Organic compounds
    • C02F2101/38—Organic compounds containing nitrogen
    • 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
    • C02F2303/04—Disinfection
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2305/00—Use of specific compounds during water treatment
    • C02F2305/06—Nutrients for stimulating the growth of microorganisms
    • 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
    • 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
    • C02F3/02—Aerobic processes
    • 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
    • C02F3/28—Anaerobic digestion processes
    • 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
    • C02F3/30—Aerobic and anaerobic processes
    • C02F3/308—Biological phosphorus removal

Abstract

The invention discloses a kind of methods of quinoline pollutant in processing sewage, including pretreatment, main process task, aeration, post-treating and other steps can remove most quinoline pollutant after treatment.The present invention has very important practical application value to the improvement of quinoline pollutant.

Description

A kind of method of quinoline pollutant in processing sewage
Technical field
The invention belongs to sewage treatment fields, and in particular to the processing of quinoline pollutant in sewage.
Background technique
Quinoline is a kind of colourless, the hygroscopic liquid with penetrating odor.Dissolve in ethyl alcohol, ether, benzene, curing Carbon is slightly soluble in water.From the point of view of its chemical structure, it belongs to heterocyclic arene type organic, by shape when coal, timber, PETROLEUM PROCESSING At.Quinoline in coking wastewater then mostly generates during decoking oil wash benzene.The pollution of quinoline is mainly by atmosphere, water body, ground The modes such as lower coal mining are propagated, also containing the ingredient of quinoline in the gas of cigarette discharge.Some researchers study discovery liver Dirty is the target organ of quinoline, is had a certain impact to liver cancer.
Quinoline is typical polycyclic aromatic nitrogen-contg jeterocyclic chemistry compound, has big toxicity, teratogenesis and carcinogenicity strong etc. special Point, it is difficult to which biodegrade can have huge potential danger to human health and ecological environment by soil pollution groundwater resources Evil, increasingly causes the concern of people.Quinoline is the Typical Representative of nitrogen-containing heterocycle compound, and purposes is extremely extensive, is a variety of doctors Important source material and solvent in the production such as medicine, agricultural product, dyestuff.But quinoline and its derivates can carcinogenic, teratogenesis, mutagenesis, And it is difficult to biodegrade, it is the severe pollutant in environment.Quinoline in environment is mainly from coal gas, fossil fuel processing, coal Tar residue and wood preservation facility.In addition, quinoline and its derivates or coking wastewater, petroleum wastewater, pharmacy waste water etc. Persistent organic pollutants ingredient in various industrial sewage.Progress is formed to the organic matter of 2 kinds of coking wastewaters respectively by scholar Analysis, it is indicated that the content of quinolines occupies the second of organic pollutant.Since quinoline contains the very strong nitrogen of I electronegativity Atom makes its water solubility enhancing, therefore this kind of compound is easier to spread in the environment and persistently exist.Have investigation discovery to exist Quinoline and its oxyquinoline at concentrations up to several mg/L in underground water and soil near creosote polluted place.Except this it Outside, quinoline and its derivates are also present in urban air, tobacco smoke, seawater and the tissue of fish, therefore are found and effectively removed The method of quinoline has a very important significance.
Now, there is the processing method of many quinoline.It is roughly divided into physical method, chemical method and bioanalysis.Physical method and change Although method removal efficiency is high, at high cost, it is easy to produce secondary pollution.Biological treating have at low cost, secondary pollution it is light, The advantages such as Environmental compatibility is good, have become one of the preferred option in pollutant abatement technology.The success of decision biochemical processing process, Effectively, applicable factor, the functional gene other than process conditions and operational administrative, for processes such as contaminant degradation, conversions Important function be also obvious.Biological treatment is because it is big with treating capacity, cost is relatively low, mild condition and does not generate The advantages that secondary pollution and have been favored by people.In recent years, pass through specific function on the basis of not changing existing treatment facility The biological reinforcing technology of the addition of energy microorganism can greatly improve original biological treatment system and imitate to the removal of target contaminant Fruit, sewage treatment capacity greatly improve, thus are increasingly valued by people.
Domestic and international researcher begins to the microorganism of separation degradation quinoline from the 1970s.Microorganism in relation to quinoline The research of degradation pathway and its degradation kinetics, science of heredity is also being goed deep into, but up to the present, the biodegrade of such compound Process study is still fairly limited, and then less for the other function research of quinoline-degrading bacterium.
Summary of the invention
Method for waste water containing quinoline is handled using biotechnology one of the objects of the present invention is to provide a kind of, to solve waste water In pollutant containing quinoline be difficult to biodegrade and the problem of detoxification.
The present invention is achieved by the following technical scheme:
A kind of processing method of the waste water containing quinoline compound, includes the following steps:
(1) it pre-processes: by filtering and precipitating, removing the suspended matter in handled sewage；
(2) main process task: addition nitrogen oxide is made into composite waste, collects in intake pool, wherein carbon-nitrogen ratio 5- 10:1, carbon in carbon-nitrogen ratio from quinoline compound, nitrogen from addition nitrogen oxide；
(3) composite waste is promoted in upflow type aerating biological filter by intake pump, oxygen supply aeration 6h；
(4) sewage after being aerated is transferred to anoxic zone, adds the floating stuffing for carrying denitrifying bacterium, handles 1 day；
(5) post-process: by treated for anoxic zone sewage it is sterile-processed after, direct emission or reuse.
Preferably, the denitrifying bacterium is to be transferred to any one following PPKL1 protein variant of coding (relative to PPKL1 Albumen original amino acid (its amino acid sequence referring to: GenBank:NP_742191.1) carry out following mutation 60E/ respectively R, 69F/P, 82P/G, 120I/F, 147L/A, 178R/S, 189E/V, 229I/Q, 243V/Q, 251S/E, 259E/P) (60E/R Indicate that in original series the 60th E amino acid substitution be R amino acid, other meanings are similar) encoding gene CCTCC M 2010209 bacterial strains.
The present invention additionally provides a kind of quinoline-degrading albumen PPKL1, amino acid sequence such as NP_742191.1.The albumen It is to be screened from pseudomonas putida by the differential protein expression that quinoline induces.The albumen has stronger removal point Solve the ability of quinoline.
The present invention provides a series of PPKL1 protein variant (relative to PPKL1 albumen original amino acid respectively into Row following mutation 60E/R, 69F/P, 82P/G, 120I/F, 147L/A, 178R/S, 189E/V, 229I/Q, 243V/Q, 251S/E, 259E/P), the ability of quinoline is decomposed with stronger removal.The coded sequence of the variant amino acids by shown in is in CCTCC M When expressing in 2010209 bacterial strains, discovery recombinant bacterium relative to the polyP bacteria of the unconverted gene there is preferably removal to decompose The ability of quinoline has better effect.
The albumen is imported into denitrifying bacterium, which not only has the effect of preferably going to denitrogenate phosphorus, while can also Quinoline in enough effective removal sewage, with good application prospect.
The above method has the advantages that
1, processing method through the invention substantially increases the degradation rate of quinoline, reduces the reaction time.
2, the albumen and conversion bacterial strain, quinoline removal efficiency with higher and nitrogen phosphorus ligands of a kind of quinoline of degrading are provided Effect.
In order to make it easy to understand, the present invention will be described in detail by specific embodiment below.It needs to refer in particular to Out, specific example is merely to illustrate, and be not meant to limit the scope of the invention.The ordinary skill people of obvious this field Member can make various modifications and variations to the present invention within the scope of the invention according to illustrating herein, these amendments It is also included in the scope of the present invention with changing.
Specific embodiment
The processing of 1 quinoline compound waste water of embodiment
(1) it pre-processes: taking and be filtered for 100mg/L sewage by conventional filtering ponds containing quinoline concentration, by normal The sedimentation basin of rule is precipitated, and the suspended matter in handled sewage is removed；
(2) main process task: will eliminate suspended matter treated sewage, addition nitrogen oxide (if nitrogen content is enough, Can also not have to addition), it is made into composite waste, collects in intake pool, wherein carbon-nitrogen ratio 8:1, the carbon in carbon-nitrogen ratio come from In quinoline compound, nitrogen from addition nitrogen oxide；
(3) composite waste is promoted in upflow type aerating biological filter by intake pump, oxygen supply aeration 6h；
(4) sewage after being aerated is transferred to anoxic zone, adds the floating stuffing for carrying denitrifying bacterium, handles 1 day；It is described anti- Nitrifier is the bacterial strain that culture presevation number is CCTCC M 2010209, specifically may refer to CN102115719A；
(5) post-process: by treated for anoxic zone sewage it is sterile-processed after, by measurement, wherein the concentration of quinoline is 0.5mg/L。
The acquisition of 2 quinoline-degrading gene PPKL1 of embodiment
Pseudomonas putida is taken, using quinoline as inducer, carries out Fiber differentiation, respectively the bacterium to induce and not induce Strain as a comparison, carries out the extraction of total protein, carries out bidirectional two-dimensional electrophoresis, passes through control, it was found that differentially expressed protein at 10. Wherein choose wherein that the biggish point of differential expression is sequenced at one.Corresponding amino acid sequence is obtained, sequence is such as Shown in Genbank:NP_742191.1.The gene is carried out to conventional prokaryotic expression, specific steps are all conventional technique.It is logical Determination of activity discovery is crossed, which has stronger quinoline removal effect.
The activity of embodiment 3PPKL1 protein mutation improves
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 are introduced between in albumen shown in NP_742191.1, from And construct different mutant proteins (preparation method that can refer to the prior art can be obtained).It prepares and obtains different prominent Then variant gene is connected respectively at carrier, the recombinant plasmid transformed being proved to be successful is entered denitrifying bacterium CCTCC M In 2010209, the genetic engineering bacterium of degradation quinoline is obtained.
The verifying of 4 quinoline-degrading effect of embodiment
Multiple engineering bacterias that embodiment 3 is obtained are distinguished according to the method for embodiment 1, respectively under different quinoline concentration The effect of the corresponding degradation quinoline of verifying.Using the Ep2 bacterium of 102206657 A of CN as control.Concrete outcome is as shown in the table:
Can be seen that the denitrifying bacterium major part for having imported the albumen PPKL1 of mutation from above result all has enhancing Removal quinoline effect, this absolutely prove is not arbitrary change all have preferable effect.

Claims (2)

1. a kind of processing method of waste water containing quinoline compound includes the following steps: that (1) pre-processes: by filtering and precipitating, going Except the suspended matter in handled sewage；(2) main process task: addition nitrogen oxide is made into composite waste, collects in intake pool, Wherein, carbon-nitrogen ratio 5-10:1, carbon in carbon-nitrogen ratio from quinoline compound, nitrogen from addition nitrogen oxide；(3) Composite waste is promoted in upflow type aerating biological filter by intake pump, oxygen supply aeration 6h；(5) sewage after being aerated is transferred to scarce Oxygen area adds the floating stuffing for carrying denitrifying bacterium, handles 1 day；(6) it post-processes: by treated for anoxic zone sewage through disappearing After poison processing, direct emission or reuse, the denitrifying bacterium are the encoding gene for being transferred to the following PPKL1 protein variant of coding 2010209 bacterial strain of CCTCC M, the PPKL1 protein variant be relative to PPKL1 albumen original amino acid carry out such as Lower mutation 189E/V, PPKL1 albumen original amino acid is referring to GenBank:NP_742191.1.
2.PPKL1 protein variant, it is characterised in that: the PPKL1 protein variant is relative to PPKL1 albumen Original amino sequence Column carry out following mutation 189E/V, and PPKL1 albumen original amino acid is referring to GenBank:NP_742191.1.
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CN201610600488.0A Active CN106045224B (en) 2015-06-05 2015-06-05 A kind of method of quinoline pollutant in processing sewage
CN201610599676.6A Active CN106007251B (en) 2015-06-05 2015-06-05 A kind of method of quinoline pollutant in processing sewage
CN201610599661.XA Active CN105967467B (en) 2015-06-05 2015-06-05 A kind of method of quinoline pollutant in treated sewage
CN201610600496.5A Active CN106116030B (en) 2015-06-05 2015-06-05 A kind of method of quinoline pollutant in treated sewage
CN201610600438.2A Active CN105948429B (en) 2015-06-05 2015-06-05 A kind of method of quinoline pollutant in treated sewage
CN201610595196.2A Active CN106007247B (en) 2015-06-05 2015-06-05 A kind of method of quinoline pollutant in treated sewage
CN201610600468.3A Active CN106145523B (en) 2015-06-05 2015-06-05 A kind of method of quinoline pollutant in processing sewage
CN201610595179.9A Active CN105948428B (en) 2015-06-05 2015-06-05 A kind of method of quinoline pollutant in processing sewage
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