CN103667161A - Pseudomonas stutzeri KY-02 and application thereof - Google Patents
Pseudomonas stutzeri KY-02 and application thereof Download PDFInfo
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- CN103667161A CN103667161A CN201310746598.4A CN201310746598A CN103667161A CN 103667161 A CN103667161 A CN 103667161A CN 201310746598 A CN201310746598 A CN 201310746598A CN 103667161 A CN103667161 A CN 103667161A
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Abstract
The invention provides pseudomonas stutzeri KY-02 which has a preservation number of CGMCC (China General Microbiological Culture Collection Center) No.8526. The pseudomonas stutzeri KY-02 can remarkably improve the efficiency of anaerobic production of marsh gas and the decomposition rate of organic matters in sludge on the basis of not influencing the stability of an anaerobic digestion reaction system, has the advantages of low cost, long duration, no secondary pollution and the like compared with chemical methods, is convenient to culture and use, can make positive contribution on further popularization and generalization of an industrial wastewater biological treatment technology, in particular to an anaerobic digestion technology and has a wide application prospect in the field of sewage treatment.
Description
Technical field
The present invention relates to biological technical field, especially a kind of Pseudomonas stutzeri KY-02 and monoclonal antibody 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., and can not be miscible with petroleum hydrocarbon.It has been widely used in 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.
Yet epoxy chloropropane is one of the national supervision of emphasis in the first batch hazardous chemical: its steam has intense stimulus to respiratory tract, repeatedly can cause lung, liver and renal impairment with long-time suction, and high density suction causes central nervous system inhibition can be lethal; Its steam has intense stimulus to eye, and liquid can cause eye and burn; The direct contact liq of skin can cause burns; Oral liver, the renal impairment of causing, can be lethal; Can there is neurasthenic syndrome and peripheral neuropathy in long-term a small amount of suction; The subcutaneous experiment of mouse shows, during minimum toxic dose 720mg/kg, just can cause tumer positive.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, can there is violent decomposition, cause that container breaks or explosion hazard.
Although epoxy chloropropane has above-mentioned many harm to the mankind and environment, along with industrial development, still there is a large amount of epoxy chloropropane to be discharged in trade effluent.This has not only improved processes load, has increased intractability, also may make the poisoning even collapse of existing Sludge System simultaneously.
Common micro-organisms and active sludge are all difficult to realize effective degraded of epoxy chloropropane.Therefore, the current treatment process for epoxy chloropropane waste water focuses mostly in chemical processes such as Fenton reagent oxidation factures.Happy report to brightness in 2008 used the processing case of Fenton reagent oxidation facture to epoxy chloropropane artificial wastewater, determined when initial pH value be 3, when adding ferrous ion and hydrogen peroxide and being respectively 1.60g/L and 1.92g/L, COD can reduce by 89.44%.Li little Lin has reported in 2009 and has adopted single factor and orthogonal test method to investigate temperature of reaction, pH value, reaction times, FeSO
4and H
2o
2the impact of the factors such as dosage on clearance, after optimal conditions, clearance is 60% left and right.Chen Xinde reported in 2012 and for certain chlor-alkali plant, in epoxy chloropropane production process, at the bottom of cyclisation tower tower, discharged the chemical Treatment of waste water, and after optimal conditions, CODCr clearance can reach 90.4%.
Yet 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 bacterial strain that can efficient degradation epoxy chloropropane, becomes one of industrial sewage process 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 to provide the application of 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, separated from active sludge, on screening flat board, show as neat, the moistening thickness of white edge, gramstaining is negative; By 16SrDNA and Biolog microorganism classification identification systems, identify, 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 V3 variable region and the V8-V9 variable region of bacterial 16 S rDNA fragment as.
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 aspect epoxy chloropropane.
Preferably, the application of above-mentioned Pseudomonas stutzeri KY-02, by described Pseudomonas stutzeri KY-02 inoculation, to LB substratum, 30 ℃ of shaking culture are dense to bacterium is O.D
600≈ 2.0, with the inoculum size of 1-5%, are inoculated in the waste water containing epoxy chloropropane, cultivate 1-3 days for 20-30 ℃, maintain dissolved oxygen more than 3.0ppm, realize 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), epoxy chloropropane that can be in efficient degradation sewage under aerobic condition, reduce COD, compare advantages such as thering is cost is low, removal efficiency is high, non-secondary pollution with chemical process, will make positive contribution for trade effluent biologic treating technique, at sewage treatment area, have wide practical use.
Accompanying drawing explanation
Fig. 1 is the systematic evolution tree of Pseudomonas stutzeri KY-02 of the present invention, and in figure, Pseudomonas stutzeri 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 and solution formula different in following embodiment are 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 that is 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, in 30 ℃ of thermostat containers, cultivate, purifying 2-3 time that repeat to rule of picking list bacterium colony, obtains purifying bacterial strain; Through 16SrDNA and Biolog microorganism classification identification systems, identify, 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:
CGGGGGGCGGCTACACATGCAAGTCGAGCGGATGAAGGGAGCTTGCTCTCTGATTCAGCGGC?GGACGGGTGAGTAATGCCTAGGAATCTGCCTGATAGTGGGGGACAACGTTTCGAAAGGAACGCTAA?TACCGCATACGTCCTACGGGAGAAAGCAGGGGACCTTCGGGCCTTGCGCTATCAGATGAGCCTAGG?TCGGATTAGCTAGTTGGTGAGGTAATGGCTCACCAAGGCGACGATCCGTAACTGGTCTGAGAGGAT?GATCAGTCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGG?ACAATGGGCGAAAGCCTGATCCAGCCATGCCGCGTGTGTGAAGAAGGTCTTCGGATTGTAAAGCAC?TTTAAGTTGGGAGGAAGGGCATTAACCTAATACGTTAGTGTTTTGACGTTACCGACAGAATAAGCA?CCGGCTAACTTCGTGCCAGCAGCCGCGGTAATACGAAGGGTGCAAGCGTTAATCGGAATTACTGGG?CGTAAAGCGCGCGTAGGTGGTTTGTTAAGTTGAATGTGAAAGCCCCGGGCTCAACCTGGGAACTGC?ATCCAAAACTGGCAAGCTAGAGTATGGCAGAGGGTGGTGGAATTTCCTGTGTAGCGGTGAAATGCG?TAGATATAGGAAGGAACACCAGTGGCGAAGGCGACCACCTGGGCTAATACTGACACTGAGGTGCGA?AAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTCGACTAGCCG?TTGGGATCCTTGAGATCTTAGTGGCGCAGCTAACGCATTAAGTCGACCGCCTGGGGAGTACGGCCG?CAAGGTTAAAACTCAAATGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGA?AGCAACGCGAAGAACCTTACCAGGCCTTGACATGCAGAGAACTTTCCAGAGATGGATTGGTGCCTT?CGGGAACTCTGACACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTC?CCGTAACGAGCGCAACCCTTGTCCTTAGTTACCAGCACGTTAAGGTGGGCACTCTAAGGAGACTGC?CGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGGCCCTTACGGCCTGGGCTACA?CACGTGCTACAATGGTCGGTACAAAGGGTTGCCAAGCCGCGAGGTGGAGCTAATCCCATAAAACCG?ATCGTAGTCCGGATCGCAGTCTGCAACTCGACTGCGTGAAGTCGGAATCGCTAGTAATCGTGAATC?AGAATGTCACGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTT?GCTCCAGAAGTAGCTAGTCTAACCTTCGGGGGGACGGTACCACGGAGATCTG
Embodiment 2
The experiment of Pseudomonas stutzeri described in embodiment 1 (Pseudomonas stutzeri) KY-02 degraded epoxy chloropropane
Processing object is screening culture medium, and operation scheme is shaking flask shaking culture.Concrete implementation step is as follows: Pseudomonas stutzeri KY-02 bacterial classification is seeded to LB substratum, and 30 ℃ of shaking culture are dense to bacterium is O.D
600≈ 2.0, with 1% inoculum size, is inoculated in liquid screening substratum, and 30 ℃ of shaking culture 1 day, maintaining dissolved oxygen is 3.0ppm; Centrifuging and taking supernatant, measures COD according to < < dichromate titration (GB11914-89) > >.Result demonstration, COD clearance is 95.6%.
Embodiment 3
The experiment of Pseudomonas stutzeri described in embodiment 1 (Pseudomonas stutzeri) KY-02 degraded epoxy chloropropane
Processing 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 bacterial classification is seeded to LB substratum, and 30 ℃ of shaking culture are dense to bacterium is O.D
600≈ 2.0, with 5% inoculum size, is inoculated in SBR system, and the lower aeration operation of room temperature (20-25 ℃) 2 days, maintaining dissolved oxygen is 3.0ppm; System stops aeration, and precipitation is got supernatant, according to < < dichromate titration (GB11914-89) > >, measures COD.Result demonstration, COD clearance is 90.4%.
Embodiment 4
The experiment of Pseudomonas stutzeri described in embodiment 1 (Pseudomonas stutzeri) KY-02 degraded epoxy chloropropane
Processing object is the pending waste water that is 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 bacterial classification is seeded to LB substratum, and 30 ℃ of shaking culture are dense to bacterium is O.D
600≈ 2.0, with 5% inoculum size, is inoculated in SBR system, and the lower aeration operation of room temperature (20-25 ℃) 3 days, maintaining dissolved oxygen is 3.5ppm; System stops aeration, and precipitation is got supernatant, according to < < dichromate titration (GB11914-89) > >, measures COD.Result demonstration, COD clearance is 92.3%.
Above-mentioned with reference to embodiment to this kind of Pseudomonas stutzeri KY-02 and the application detailed description of carrying out; illustrative rather than determinate; can list several embodiment according to institute's limited range; therefore in the variation and the modification that do not depart under general plotting of the present invention, within should belonging to 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: separation, from active sludge, shows as neat, the moistening thickness of white edge on screening flat board, and gramstaining is negative; By 16SrDNA and Biolog microorganism classification identification systems, identify, 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 described Pseudomonas stutzeri KY-02 of one of claim 1-3, is characterized in that: the application in degraded trade effluent aspect 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, to LB substratum, 30 ℃ of shaking culture are dense to bacterium is O.D
600≈ 2.0, with the inoculum size of 1-5%, are inoculated in the waste water containing epoxy chloropropane, cultivate 1-3 days for 20-30 ℃, maintain dissolved oxygen more than 3.0ppm, realize the degraded to epoxy chloropropane.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105969760A (en) * | 2016-06-29 | 2016-09-28 | 华南理工大学 | Embedded nano-iron/single-microorganism composite microbial agent and preparation method thereof |
| CN106630173A (en) * | 2015-11-04 | 2017-05-10 | 中国石油化工股份有限公司 | Biochemical treatment method of epoxy chloropropane waste water |
| 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 |
| CN113801828A (en) * | 2021-11-05 | 2021-12-17 | 黄河三角洲京博化工研究院有限公司 | Efficient nitrobenzene degrading bacterium and preparation and application thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106630173A (en) * | 2015-11-04 | 2017-05-10 | 中国石油化工股份有限公司 | Biochemical treatment method of epoxy chloropropane waste water |
| 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 |
| CN105969760A (en) * | 2016-06-29 | 2016-09-28 | 华南理工大学 | Embedded nano-iron/single-microorganism composite microbial agent and preparation method thereof |
| 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 |
| CN113801828A (en) * | 2021-11-05 | 2021-12-17 | 黄河三角洲京博化工研究院有限公司 | Efficient nitrobenzene degrading bacterium and preparation and application 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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