CN102424806A - DDT (dichlorodiphenyltrichloroethane)-degrading pseudomonad strain - Google Patents
DDT (dichlorodiphenyltrichloroethane)-degrading pseudomonad strain Download PDFInfo
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- CN102424806A CN102424806A CN2011104209991A CN201110420999A CN102424806A CN 102424806 A CN102424806 A CN 102424806A CN 2011104209991 A CN2011104209991 A CN 2011104209991A CN 201110420999 A CN201110420999 A CN 201110420999A CN 102424806 A CN102424806 A CN 102424806A
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Abstract
The invention relates to a DDT (dichlorodiphenyltrichloroethane)-degrading pseudomonad strain, and specifically provides a pseudomonad strain capable of degrading DDT, which can be used for the biological intensified treatment of DDT pollutants and can help to degrade DDT organochlorine pesticides. The DDT-degrading pseudomonad strain is named pseudomonad No.12-3; the strain was preserved by China General Microbiological Culture Collection Center on November 8th, 2011; and the preservation number is CGMCC NO.5449. The pseudomonad No.12-3 has the characteristic of degrading DDT organochlorine pesticides; the DDT degradation rate can be up to 51.6%; and secondary pollution can not be caused to the environment. Thus, the DDT-degrading pseudomonad strain can help to solve the problems of DDT degradation, pollution remediation and the like, thereby having high application value.
Description
Technical field
The present invention relates to the pseudomonas of a kind of DDT of degraded.
Background technology
DDT is DDT again; Chemistry biconjugate chloro-phenyl-trichloroethane (Dichlorodiphenyltrichloroethane) by name; 12 kinds that are " about the Convention of Stockholm of persistence organic pollutant POPs " regulation limit one of POPs, are a kind of heat-resisting, acidproof, fat-soluble big, organochlorine class broad spectrum pesticides that the longevity of residure is long.Because it has the characteristics of teratogenesis, carcinogenic, mutagenic " three cause " effect, high toxicity and difficult degradation, and environment has been caused serious pollution.Though forbidden many decades so far, the accumulative total effect in environment is still very serious.Therefore, the environmental problem that adopts certain technique means reparation to have the organochlorine pesticide of the DDT class of high poison, high residue, difficult degradation to cause is extremely urgent.
Microbiological deterioration be considered to organochlorine pesticide degraded the most reliably, approach the most efficiently.The mikrobe of separation screening efficient degradation DDT is the effective measure that people carry out DDT Pollution abatement, soil organisms reparation.Through screening DDT degradation bacteria, study its biological characteristics, degradation characteristic and DDT degradation bacteria Optimizing Conditions of Fermentation thereof, the biological prosthetic that can be the DDT contaminated soil provides foundation, reaches to recover the soil organisms variety, the purpose of protection environment and human health.
Summary of the invention
The invention provides a pseudomonas bacterial strain, degradable DDT can be applicable to the biological reinforced processing of DDT pollutent, for degraded DDT class organochlorine insecticide is offered help.
The degrade pseudomonas of DDT of the present invention is pseudomonas (Pseudomonas sp.) 12-3#; Be deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC); The preservation address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City; Preservation date is on November 08th, 2011, and preserving number is CGMCC No.5449; Pseudomonas 12-3# of the present invention is a gram-positive microorganism, and aerobic bacteria is bar-shaped, and the thalline size is (0.1 μ m～0.2 μ m) * (0.4 μ m～0.6 μ m), amphitrichous; On the solid inorganic salt culture medium, cultivate, bacterium colony is rounded, and color is faint yellow, transparent, smooth surface, and neat in edge, thickness is prone to provoke.
Pseudomonas 12-3# of the present invention can not utilize glucose, can not oxidation ethanol or aldose.The catalase reaction is positive, and produces indole test and is positive, and produces ammonia test and is positive; The methyl red test reaction is negative, and the starch hydrolysis is negative, and fat hydrolysis is negative; The urea experiment is negative, and the PD experiment is negative, and the glucose fermentation test is negative.
Pseudomonas 12-3# of the present invention can grow on the minimal medium that has added DDT, and the best final concentration of DDT is 20mg/L in the substratum.The optimum growth temperature of pseudomonas 12-3# of the present invention is 30 ℃, and the pH value of suitable growing environment is 7.0.
Pseudomonas of the present invention (Pseudomonas sp.) 12-3# identifies through lipid acid; (Pseudomonas) is the most close with Rhodopseudomonas; Confirm that pseudomonas 12-3# is the novel bacterial of Rhodopseudomonas (Pseudomonas), called after pseudomonas 12-3# through combining morphological features, growth conditions, Physiology and biochemistry qualification result.
Pseudomonas 12-3# of the present invention has the characteristic of degraded DDT class organochlorine insecticide, can reach 51.6% to the degradation rate of DDT, does not cause secondary environmental pollution.Pseudomonas 12-3# of the present invention is helpful for solving problems such as DDT degraded and pollution reparation, has higher using value.
Pseudomonas of the present invention (Pseudomonas sp.) 12-3# belongs to Rhodopseudomonas (Pseudomonas); Be deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC); The preservation address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City; Preservation date is on November 08th, 2011, and preserving number is CGMCC No.5449.
Description of drawings
Fig. 1 is the dyeing picture of pseudomonas 12-3# of the present invention; Fig. 2 is a pseudomonas 12-3# atom mechanics microscope aspect graph of the present invention; Fig. 3 is that the lipid acid of pseudomonas 12-3# is identified collection of illustrative plates.
Embodiment
Technical scheme of the present invention is not limited to following cited embodiment, also comprises the arbitrary combination between each embodiment.
Embodiment one: the pseudomonas of this embodiment degraded DDT is pseudomonas (Pseudomonas sp.) 12-3#; Be deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC); The preservation address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City; Preservation date is on November 08th, 2011, and preserving number is CGMCCNo.5449.
This embodiment pseudomonas (Pseudomonas sp.) 12-3# is a gram-positive microorganism; Aerobic bacteria; Bar-shaped, the thalline size is (0.1 μ m～0.2 μ m) * (0.4 μ m～0.6 μ m), amphitrichous; The dyeing picture of pseudomonas 12-3# is as shown in Figure 1, and pseudomonas 12-3# atom mechanics microscope aspect graph is as shown in Figure 2; On the solid inorganic salt culture medium, cultivate, bacterium colony is rounded, and color is faint yellow, transparent, smooth surface, and neat in edge, thickness is prone to provoke.Wherein said solid inorganic salt culture medium (1000mL) is by 4.0g NaNO
3, 1.5g KH
2PO
4, 0.5g Na
2HPO
4, 0.005g FeCl
3, 0.01g CaCl
2, 20g agar powder and 20mg/L DDT form.
With reference to " the outstanding Bacteria Identification handbook of uncle the 8th edition is carried out Physiology and biochemistry with " common bacteria system identification handbook " to pseudomonas (Pseudomonas sp.) 12-3# and is identified: can not utilize glucose, and can not oxidation ethanol or aldose.The catalase reaction is positive, and produces indole test and is positive, and produces ammonia test and is positive; The methyl red test reaction is negative, and the starch hydrolysis is negative, and fat hydrolysis is negative; The urea experiment is negative, and the PD experiment is negative, and the glucose fermentation test is negative.
This embodiment pseudomonas 12-3# can grow on the minimal medium that has added DDT, and the best final concentration of DDT is 20mg/L in the substratum.The minimal medium that has wherein added DDT is by 4.0g NaNO
3, 1.5g KH
2PO
4, 0.5g Na
2HPO
4, 0.005g FeCl
3, 0.01g CaCl
2Form with the DDT of 20mg/L.The optimum growth temperature of pseudomonas 12-3# of the present invention is 30 ℃, and the pH value of suitable growing environment is 7.0.
Embodiment two: this embodiment pseudomonas (Pseudomonas sp.) 12-3# is received to screen in the DDT Contaminated soil by the Guangzhou and obtains.Screening is carried out according to the following steps: 1. get 10mL and insert in the sterilized Erlenmeyer flask that granulated glass sphere and 90mL sterilized water be housed through the soil suspension liquid of DDT domestication and fully vibrate, to break up zoogloea, make bacterium be unicellular state and be scattered in the liquid; 2. get the 1mL bacteria suspension the soil bacteria suspension in Erlenmeyer flask, being inoculated in DDT with coubling dilution is in the solid inorganic salt culture medium of sole carbon source, is inverted in 30 ℃ of constant incubators and cultivates 3d; 3. distinguish the different typical single bacterium colony of picking, marked on plate, and streak culture in new petridish, repeat above operation 3～4 times, obtain the consistent pure bacterium flat board of colony characteristics; 4. the 4 ℃ of preservations of transferring in preserving substratum of the pure bacterium of gained are subsequent use; Carrying out multiple sieve then, promptly is that on the basis of primary dcreening operation, further whether to have higher DDT degradation rate with bacterial strain be foundation, filters out degradation efficiency bacterial strain preferably, is pseudomonas 12-3#.
Every liter of solid inorganic salt culture medium is by 4.0g NaNO in this embodiment
3, 1.5g KH
2PO
4, 0.5g Na
2HPO
4, 0.005g FeCl
3, 0.01g CaCl
2, 20g agar powder and 20mg/L DDT form, the pH value is 7.0～7.2,121 ℃ of autoclaving 20min.
Adopt vapor-phase chromatography to detect pseudomonas 12-3# cell fatty acid and carry out strain identification.Utilize gc to short chain fatty acid specific among the pseudomonas 12-3# (C9～C20) analyze; The lipid acid of pseudomonas 12-3# identifies that collection of illustrative plates is as shown in Figure 3; Compare according to the kind of short chain fatty acid and the collection of illustrative plates of content and built-in DB TSBA66.00; It is the most approaching that the result shows that 12-3# fatty acid structure and pseudomonas vesicularis belong to, and similarity is 64.28%, confirms that finally pseudomonas 12-3# belongs to Rhodopseudomonas (Pseudomonas).
Adopt the degradation rate of this bacterial strain of high effective liquid chromatography for measuring to DDT.This bacterial strain bacteria suspension 1mL is inoculated in 50mL has added that (minimal medium that has wherein added DDT is by 4.0g NaNO in the minimal medium of DDT
3, 1.5gKH
2PO
4, 0.5g Na
2HPO
4, 0.005g FeCl
3, 0.01g CaCl
2Form with the DDT of 20mg/L), under 30 ℃, 120r/min condition, cultivate 8d, bacteria suspension is through centrifugal; N-hexane extraction, anhydrous sodium sulfate dehydration, Rotary Evaporators concentrates; After measure the whole content of DDT with high performance liquid chromatograph behind the normal hexane constant volume, final DDT degradation rate reaches 51.6%.Proof pseudomonas 12-3# has the function of effective degraded DDT, and need not to add other chemicals, and DDT is polluted to repair provides strong theoretical foundation, has broad application prospects.
Claims (1)
1. the pseudomonas of strain degraded DDT; The pseudomonas of DDT of it is characterized in that degrading is pseudomonas (Pseudomonas sp.) 12-3#; Be deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center; The preservation address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, and preservation date is on November 08th, 2011, and preserving number is CGMCCNo.5449.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103223219A (en) * | 2013-04-11 | 2013-07-31 | 中国科学院水生生物研究所 | CYP5013C2 protein and coding gene thereof, and tetrahymena expressing CYP5013C2 protein |
| CN103243057A (en) * | 2013-05-27 | 2013-08-14 | 江苏南资环保科技有限公司 | Pseudomonas SYA-1 for degrading triethylamine and application of pseudomonas SYA-1 |
| CN106336082A (en) * | 2016-10-20 | 2017-01-18 | 辽宁海润环保技术股份有限公司 | Method for treating fungicide production waste water |
| CN107099391A (en) * | 2017-05-10 | 2017-08-29 | 颐中（青岛）实业有限公司 | It is a kind of to be used to remove fruit and vegetable surfaces DDT microbial bacterial agent and preparation method thereof |
| CN108130281A (en) * | 2018-01-30 | 2018-06-08 | 陕西师范大学 | Aspergillus terreus DDT98801 and its screening technique and the application in degradation of dichloro-diphenyl-trichloroethane |
| CN108452666A (en) * | 2018-05-14 | 2018-08-28 | 大江环境股份有限公司 | A kind of application of the biological active filling material comprising Meng Shi pseudomonads in purifying chlorine contained exhaust gas |
| CN109622608A (en) * | 2018-12-31 | 2019-04-16 | 陕西师范大学 | Application of one plant of branch acremonium bacteria strain Y0997 in processing soil and water pollution |
| CN110438031A (en) * | 2019-06-28 | 2019-11-12 | 中南大学 | A kind of microbial deoderizer and preparation method thereof |
| CN116396898A (en) * | 2023-03-10 | 2023-07-07 | 江苏诚冉环境修复工程有限公司 | 1, 2-trichloroethane degrading bacterium and application thereof |
| CN116673319A (en) * | 2023-07-31 | 2023-09-01 | 北京建工环境修复股份有限公司 | Degradation method of polystyrene in soil |
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Patent Citations (1)
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| CHANDRAPPA M. KAMANAVALLI ET AL.: "Biodegradation of DDT by a Pseudomonas Species.", 《CURRENT MICROBIOLOGY》 * |
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Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103223219A (en) * | 2013-04-11 | 2013-07-31 | 中国科学院水生生物研究所 | CYP5013C2 protein and coding gene thereof, and tetrahymena expressing CYP5013C2 protein |
| CN103223219B (en) * | 2013-04-11 | 2015-11-25 | 中国科学院水生生物研究所 | The tetrahymena of CYP5013C2 albumen and encoding gene and expression CYP5013C2 albumen |
| CN103243057A (en) * | 2013-05-27 | 2013-08-14 | 江苏南资环保科技有限公司 | Pseudomonas SYA-1 for degrading triethylamine and application of pseudomonas SYA-1 |
| CN103243057B (en) * | 2013-05-27 | 2015-03-25 | 江苏南资环保科技有限公司 | Pseudomonas SYA-1 for degrading triethylamine and application of pseudomonas SYA-1 |
| CN106336082A (en) * | 2016-10-20 | 2017-01-18 | 辽宁海润环保技术股份有限公司 | Method for treating fungicide production waste water |
| CN107099391A (en) * | 2017-05-10 | 2017-08-29 | 颐中（青岛）实业有限公司 | It is a kind of to be used to remove fruit and vegetable surfaces DDT microbial bacterial agent and preparation method thereof |
| CN108130281A (en) * | 2018-01-30 | 2018-06-08 | 陕西师范大学 | Aspergillus terreus DDT98801 and its screening technique and the application in degradation of dichloro-diphenyl-trichloroethane |
| CN108452666A (en) * | 2018-05-14 | 2018-08-28 | 大江环境股份有限公司 | A kind of application of the biological active filling material comprising Meng Shi pseudomonads in purifying chlorine contained exhaust gas |
| CN108452666B (en) * | 2018-05-14 | 2019-03-19 | 大江环境股份有限公司 | A kind of application of the biological active filling material comprising Meng Shi pseudomonad in purification chlorine contained exhaust gas |
| CN109622608A (en) * | 2018-12-31 | 2019-04-16 | 陕西师范大学 | Application of one plant of branch acremonium bacteria strain Y0997 in processing soil and water pollution |
| CN109622608B (en) * | 2018-12-31 | 2021-06-15 | 陕西师范大学 | Application of acremonium cladonioides strain Y0997 in treatment of soil and water pollution |
| CN110438031A (en) * | 2019-06-28 | 2019-11-12 | 中南大学 | A kind of microbial deoderizer and preparation method thereof |
| CN116396898A (en) * | 2023-03-10 | 2023-07-07 | 江苏诚冉环境修复工程有限公司 | 1, 2-trichloroethane degrading bacterium and application thereof |
| CN116673319A (en) * | 2023-07-31 | 2023-09-01 | 北京建工环境修复股份有限公司 | Degradation method of polystyrene in soil |
| CN116673319B (en) * | 2023-07-31 | 2023-10-27 | 北京建工环境修复股份有限公司 | Degradation method of polystyrene in soil |
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Application publication date: 20120425 |