CN103667161B - A kind of Pseudomonas stutzeri KY-02 and application - Google Patents
A kind of Pseudomonas stutzeri KY-02 and application Download PDFInfo
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- CN103667161B CN103667161B CN201310746598.4A CN201310746598A CN103667161B CN 103667161 B CN103667161 B CN 103667161B CN 201310746598 A CN201310746598 A CN 201310746598A CN 103667161 B CN103667161 B CN 103667161B
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Abstract
The invention provides a kind of Pseudomonas stutzeri KY-02, does is preserving number CGMCC? No.8526, described Pseudomonas stutzeri KY-02 can on the basis not affecting anaerobic digestion reaction system stability, significantly improve efficiency and the mud organic matter decomposition rate of anaerobic methane production, there is the advantages such as cost is low, the time length of a specified duration, non-secondary pollution compared with chemical process, cultivate easy to use, can be trade effluent biologic treating technique, especially the further genralrlization of anaerobic digestion techniques makes positive contribution with universal, is with a wide range of applications at sewage treatment area.
Description
Technical field
The present invention relates to biological technical field, especially a kind of Pseudomonas stutzeri KY-02 and monoclonal antibody thereof and application.
Background technology
Epoxy chloropropane is a kind of important Organic Chemicals and fine chemical product, and in epoxy compounds, its output is only second to oxyethane and propylene oxide.It is colourless liquid, has like chloroform smell, volatile, unstable, can be miscible, water insoluble with ethanol, ether, chloroform, trieline and tetracol phenixin etc., can not be miscible with petroleum hydrocarbon.It has been widely used in the industrial production, mainly for the preparation of glycerine, epoxy resin, chlorohydrin rubber, polyether glycol, it is the important source material of producing glycerine and glycidyl derivatives, can be used as organic solvent (solvents of resin, natural gum, cellulose ester, ether of cellulose etc.), or the intermediate of rodenticide gliftor.
But epoxy chloropropane is country's one of emphasis supervision hazardous chemical in the first batch: its steam has intense stimulus to respiratory tract, and repeatedly can cause lung, liver and renal impairment with long-time suction, high density suction causes central nervous system impression can be lethal; Its steam has intense stimulus to eye, and liquid can cause eye and burn; Direct skin contact liquid can cause and burn; Orally cause liver, renal impairment, can be lethal; Neurasthenic syndrome and peripheral neuropathy can be there is in long-term a small amount of suction; The subcutaneous experiment display of mouse, just can cause tumer positive during minimum toxic dose 720mg/kg.Its aerial tolerance is 2ppm, and its steam can form explosive mixture with air, and chance naked light, high temperature can cause decomposition explosion and burning; If meet high heat violent decomposition can occur, cause vessel breaks or explosion hazard.
Although epoxy chloropropane has above-mentioned many harm to the mankind and environment, along with industrial development, a large amount of epoxy chloropropane is still had to be discharged in trade effluent.Which not only improves processing load, add intractability, also may make that existing Sludge System is poisoning even collapses simultaneously.
Common micro-organisms and active sludge are all difficult to the effective degraded realizing epoxy chloropropane.Therefore, focus mostly in chemical processes such as Fenton reagent oxidizing treatment for the treatment process of epoxy chloropropane waste water at present.Happy reporting to brightness in 2008 uses Fenton reagent oxidizing treatment to the process case of epoxy chloropropane artificial wastewater, determine when initial pH value be 3, add ferrous ion and hydrogen peroxide be respectively 1.60g/L and 1.92g/L time, COD can reduce by 89.44%.Li little Lin reported in 2009 and adopts single factor test and orthogonal test method to investigate temperature of reaction, pH value, reaction times, FeSO
4and H
2o
2the factors such as dosage are on the impact of clearance, and after optimal conditions, clearance is about 60%.Chen Xinde reported the chemical Treatment of discharging waste water for certain chlor-alkali plant at the bottom of the cyclized by treatment tower tower of epoxy chloropropane production process in 2012, after optimal conditions, CODCr clearance can reach 90.4%.
But chemical process can not obtain desirable removal efficiency, and need to add a large amount of medicaments, cost is higher, also easily causes secondary pollution.Therefore, screening obtains can the bacterial strain of efficient degradation epoxy chloropropane, becomes one of Industrial Waste Water Treatments field difficult problem urgently to be resolved hurrily.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of Pseudomonas stutzeri KY-02, has extremely strong degraded epoxy chloropropane ability.
Another technical problem to be solved by this invention is the application providing above-mentioned Pseudomonas stutzeri KY-02.
For solving the problems of the technologies described above, technical scheme of the present invention is:
A kind of Pseudomonas stutzeri (Pseudomonas stutzeri) KY-02, its preserving number is CGMCC No.8526(depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, preservation date: on December 9th, 2013, deposit number: CGMCC No.8526).
Preferably, above-mentioned Pseudomonas stutzeri KY-02, is separated from active sludge, and screening flat board shows as neat, the moistening thickness of white edge, and gramstaining is negative; Identified by 16SrDNA and Biolog microorganism classification identification systems, be defined as Pseudomonas stutzeri, called after Pseudomonas stutzeri KY-02, as shown in Figure 1,16SrDNA the sequencing results is sequence shown in <400>1 in sequence table to its systematic evolution tree.
Preferably, above-mentioned Pseudomonas stutzeri KY-02, described nucleotides sequence is classified as V3 variable region and the V8-V9 variable region of bacterial 16 S rDNA fragment.
Preferably, above-mentioned Pseudomonas stutzeri KY-02, has extremely strong degraded epoxy chloropropane ability.
Above-mentioned Pseudomonas stutzeri KY-02, the application in degraded trade effluent in epoxy chloropropane.
Preferably, the application of above-mentioned Pseudomonas stutzeri KY-02, by described Pseudomonas stutzeri KY-02 inoculation in LB substratum, 30 DEG C of shaking culture are dense to bacterium is O.D
600≈ 2.0, is inoculated in the waste water containing epoxy chloropropane with the inoculum size of 1-5%, cultivates 1-3 days for 20-30 DEG C, maintains dissolved oxygen at more than 3.0ppm, realizes the degraded to epoxy chloropropane.
The invention has the beneficial effects as follows:
The invention provides Pseudomonas stutzeri KY-02(preserving number is CGMCC No.8526), can epoxy chloropropane in efficient degradation sewage under aerobic condition, reduce COD, there is the advantages such as cost is low, removal efficiency is high, non-secondary pollution compared with chemical process, positive contribution will be made for trade effluent biologic treating technique, have wide practical use at sewage treatment area.
Accompanying drawing explanation
Fig. 1 is the systematic evolution tree of Pseudomonas stutzeri KY-02 of the present invention, and in figure, Pseudomonasstutzeri is Pseudomonas stutzeri.
Preservation information
Classification noun: Pseudomonas stutzeri (Pseudomonas stutzeri)
Depositary institution's title: China Committee for Culture Collection of Microorganisms's common micro-organisms center
Depositary institution address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City
Preservation date: on December 9th, 2013
Preserving number: CGMCC No.8526
Embodiment
Below in conjunction with specific embodiment, technical scheme of the present invention is further described.
Substratum different in following embodiment and solution formula as follows:
1. screening culture medium: epoxy chloropropane 3g, NaNO
32g, KH
2pO
40.5g, K
2hPO
40.5g, MgSO
40.1g, CaCl
20.1g, distilled water 1000mL, pH7.0.
2.LB substratum: distilled water 1000mL, peptone 10.0g, yeast powder 5.0g, NaCl5.0g, pH7.0-7.2.
3. artificial wastewater: tap water 1000mL, NaNO
30.5g, epoxy chloropropane 1g, KH
2pO
40.05g, pH6.5-7.5.
4. epoxy chloropropane wastewater from chemical industry: the pending waste water being taken from Hangu, Tianjin chemical industrial park sewage work.
Embodiment 1
The screening and identification of Pseudomonas stutzeri (Pseudomonas stutzeri) KY-02
Experiment material is from the active sludge of Hangu, Tianjin chemical industrial park sewage work biological treatment tank.
Get mud diluent 0.1ml, coat on screening culture medium flat board, cultivate in 30 DEG C of thermostat containers, picking list bacterium colony repeats line purifying 2-3 time, obtains purifying bacterial strain; Through the qualification of 16SrDNA and Biolog microorganism classification identification systems, be defined as Pseudomonas stutzeri, as shown in Figure 1, by its 16SrDNA gene of pcr amplification, sequencing result is as follows for systematic evolution tree:
CGGGGGGCGGCTACACATGCAAGTCGAGCGGATGAAGGGAGCTTGCTCTCTGATTCAGCGGCGGACGGGTGAGTAATGCCTAGGAATCTGCCTGATAGTGGGGGACAACGTTTCGAAAGGAACGCTAATACCGCATACGTCCTACGGGAGAAAGCAGGGGACCTTCGGGCCTTGCGCTATCAGATGAGCCTAGGTCGGATTAGCTAGTTGGTGAGGTAATGGCTCACCAAGGCGACGATCCGTAACTGGTCTGAGAGGATGATCAGTCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGCGAAAGCCTGATCCAGCCATGCCGCGTGTGTGAAGAAGGTCTTCGGATTGTAAAGCACTTTAAGTTGGGAGGAAGGGCATTAACCTAATACGTTAGTGTTTTGACGTTACCGACAGAATAAGCACCGGCTAACTTCGTGCCAGCAGCCGCGGTAATACGAAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCGCGTAGGTGGTTTGTTAAGTTGAATGTGAAAGCCCCGGGCTCAACCTGGGAACTGCATCCAAAACTGGCAAGCTAGAGTATGGCAGAGGGTGGTGGAATTTCCTGTGTAGCGGTGAAATGCGTAGATATAGGAAGGAACACCAGTGGCGAAGGCGACCACCTGGGCTAATACTGACACTGAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTCGACTAGCCGTTGGGATCCTTGAGATCTTAGTGGCGCAGCTAACGCATTAAGTCGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGCCTTGACATGCAGAGAACTTTCCAGAGATGGATTGGTGCCTTCGGGAACTCTGACACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGTAACGAGCGCAACCCTTGTCCTTAGTTACCAGCACGTTAAGGTGGGCACTCTAAGGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGGCCCTTACGGCCTGGGCTACACACGTGCTACAATGGTCGGTACAAAGGGTTGCCAAGCCGCGAGGTGGAGCTAATCCCATAAAACCGATCGTAGTCCGGATCGCAGTCTGCAACTCGACTGCGTGAAGTCGGAATCGCTAGTAATCGTGAATCAGAATGTCACGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTGCTCCAGAAGTAGCTAGTCTAACCTTCGGGGGGACGGTACCACGGAGATCTG
Embodiment 2
The experiment of Pseudomonas stutzeri described in embodiment 1 (Pseudomonas stutzeri) KY-02 degraded epoxy chloropropane
Handling object is screening culture medium, and operation scheme is that shake flask is cultivated.Concrete implementation step is as follows: Pseudomonas stutzeri KY-02 strain inoculation is to LB substratum, and 30 DEG C of shaking culture are dense to bacterium is O.D
600≈ 2.0, is inoculated in liquid screening medium with 1% inoculum size, 30 DEG C of shaking culture 1 day, and maintenance dissolved oxygen is 3.0ppm; Centrifuging and taking supernatant, measures COD according to " dichromate titration (GB11914-89) ".Result shows, and COD clearance is 95.6%.
Embodiment 3
The experiment of Pseudomonas stutzeri described in embodiment 1 (Pseudomonas stutzeri) KY-02 degraded epoxy chloropropane
Handling object is artificial wastewater, and operation scheme is that in 3L SBR system, aeration is cultivated.Concrete implementation step is as follows: Pseudomonas stutzeri KY-02 strain inoculation is to LB substratum, and 30 DEG C of shaking culture are dense to bacterium is O.D
600≈ 2.0, is inoculated in SBR system with 5% inoculum size, and under room temperature (20-25 DEG C), aeration runs 2 days, and maintenance dissolved oxygen is 3.0ppm; System stops aeration, and precipitation gets supernatant, measures COD according to " dichromate titration (GB11914-89) ".Result shows, and COD clearance is 90.4%.
Embodiment 4
The experiment of Pseudomonas stutzeri described in embodiment 1 (Pseudomonas stutzeri) KY-02 degraded epoxy chloropropane
Handling object is the pending waste water being taken from Hangu, Tianjin chemical industrial park sewage work, and wherein COD is 213.6ppm, and main organism is epoxy chloropropane.Operation scheme is that in 3L SBR system, aeration is cultivated.Concrete implementation step is as follows: Pseudomonas stutzeri KY-02 strain inoculation is to LB substratum, and 30 DEG C of shaking culture are dense to bacterium is O.D
600≈ 2.0, is inoculated in SBR system with 5% inoculum size, and under room temperature (20-25 DEG C), aeration runs 3 days, and maintenance dissolved oxygen is 3.5ppm; System stops aeration, and precipitation gets supernatant, measures COD according to " dichromate titration (GB11914-89) ".Result shows, and COD clearance is 92.3%.
Above-mentioned detailed description of this kind of Pseudomonas stutzeri KY-02 being carried out with application with reference to embodiment; illustrative instead of determinate; several embodiments can be listed according to institute's limited range; therefore in the change do not departed under general plotting of the present invention and amendment, should belong within protection scope of the present invention.
Claims (5)
1. a Pseudomonas stutzeri KY-02, is characterized in that: its preserving number is CGMCC No.8526.
2. Pseudomonas stutzeri KY-02 according to claim 1, is characterized in that: be separated from active sludge, and screening flat board shows as neat, the moistening thickness of white edge, and gramstaining is negative; By the qualification of 16SrDNA and Biolog microorganism classification identification systems, be defined as Pseudomonas stutzeri, called after Pseudomonas stutzeri KY-02,16SrDNA the sequencing results is sequence shown in <400>1 in sequence table.
3. Pseudomonas stutzeri KY-02 according to claim 1, is characterized in that: have extremely strong degraded epoxy chloropropane ability.
4. the application of the Pseudomonas stutzeri KY-02 that one of claim 1-3 is described in degraded trade effluent in epoxy chloropropane.
5. the application of Pseudomonas stutzeri KY-02 according to claim 4, is characterized in that: by described Pseudomonas stutzeri KY-02 inoculation in LB substratum, and 30 DEG C of shaking culture are dense to bacterium is O.D
600≈ 2.0, is inoculated in the waste water containing epoxy chloropropane with the inoculum size of 1-5%, cultivates 1-3 days for 20-30 DEG C, maintains dissolved oxygen at more than 3.0ppm, realizes the degraded to epoxy chloropropane.
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| CN106630174A (en) * | 2015-11-04 | 2017-05-10 | 中国石油化工股份有限公司 | Method for treating epichlorohydrin wastewater |
| CN106630173B (en) * | 2015-11-04 | 2019-09-10 | 中国石油化工股份有限公司 | A kind of biochemical processing method of epoxychloropropane waste water |
| CN105969760B (en) * | 2016-06-29 | 2020-02-18 | 华南理工大学 | Embedded nano iron/single microorganism composite microbial agent and preparation method thereof |
| CN113801828B (en) * | 2021-11-05 | 2023-05-12 | 黄河三角洲京博化工研究院有限公司 | High-efficiency nitrobenzene degrading bacterium and preparation and application thereof |
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| CN102168041A (en) * | 2010-12-22 | 2011-08-31 | 浙江工业大学 | Fetor pseudomonas putida ZJB09102 and application thereof in preparation of epoxy chloropropane |
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| CN102168041A (en) * | 2010-12-22 | 2011-08-31 | 浙江工业大学 | Fetor pseudomonas putida ZJB09102 and application thereof in preparation of epoxy chloropropane |
Non-Patent Citations (2)
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|---|
| Characterization of the epoxide hydrolase from an epichlorohydrin-degrading Pseudomonas sp.;Mariken H. J. JACOBS,ET AL;《Eur. J. Biochem.》;19911231;第202卷;1217－1222 * |
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