CN103820370B - A kind of Pseudomonas aeruginosa and application thereof - Google Patents
A kind of Pseudomonas aeruginosa and application thereof Download PDFInfo
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- CN103820370B CN103820370B CN201410083918.7A CN201410083918A CN103820370B CN 103820370 B CN103820370 B CN 103820370B CN 201410083918 A CN201410083918 A CN 201410083918A CN 103820370 B CN103820370 B CN 103820370B
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Abstract
The invention discloses a kind of Pseudomonas aeruginosa, does is its Classification And Nomenclature Pseudomonas aeruginosa (Pseudomonas? aeruginosa) DSP-05, be deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, does is deposit number: CGMCC? No.8390, preservation date is on October 24th, 2013.A kind of Pseudomonas aeruginosa of the present invention is mainly used in the Chlorpyrifos 94 in degrading waste water, is adapted at promoting the use of in the wastewater treatment containing Chlorpyrifos 94.And the chlorpyrifos degrading effect that employing bacterial strain is prepared into immobilized spherule is better, and it is low to have production cost, easy to use, the advantage that removal effect is good, the present invention is for preserving the ecological environment, and the protection mankind's is healthy, reduces cost for wastewater treatment and has great importance; A kind of Pseudomonas aeruginosa (Pseudomonas of the present invention? aeruginosa) DSP-05 culture condition simply, is easily preserved, and is easy to suitability for industrialized production, has good development prospect.
Description
Technical field
The invention belongs to environment and biological technical field, particularly relate to a kind of Pseudomonas aeruginosa (Pseudomonasaeruginosa) DSP-05 and the application in the degraded of Chlorpyrifos 94 factory effluent Chlorpyrifos thereof.
Background technology
Organophosphorus pesticide (OrganophosphatepesticidesOPs) is agriculturally being widely used as sterilant, is the primary categories in current agricultural chemicals, at home and abroad produces in a large number for a long time and widely uses.Organophosphorus pesticide has mutagenicity and teratogenecity, and mammiferous nerve is as mostly relevant with it in mad cow disease, Creutzfeldt-Jakob disease, the complexities of the Gulf War, parkinsonism etc. with disease of immune system.Remain in organophosphorus pesticide on veterinary antibiotics and animal food due to the function of the cruel enzyme of second phthalein choline in human body can be suppressed thus there is the different toxicity of degree to people, wherein chronic toxicity can induce polyneuropathy, apoplexy etc., and acute poisoning can cause people's muscle spasm, pupil contraction, expiratory dyspnea until death.
Chlorpyrifos 94 is a kind of efficient, broad-spectrum organophosphorous pesticide, have insect tag, stomach toxicity and fumigation action, especially have extraordinary effect to the control of brown paddy plant hopper.It is one of most widely used five kinds of desinsections in the whole world at present.Due to its biological cylinder accumulation effect, damaging effect can be produced to the environment of surrounding, marine organism and the mankind.All can contain the former medicine of different concns in the waste water produced in China's pesticide producing Chlorpyrifos 94 process, cause waste water to be difficult to carry out biological degradation.In the field application of Chlorpyrifos 94, also have and can be scattered in farmland ecosystem greatly, affect Agro-ecology balance.Microbiological deterioration has the advantages such as cost is low, efficiency is high, non-secondary pollution, ecological recovery are good, has been applied in a lot, is significant so filter out chlorpyrifos degrading bacteria for process Chlorpyrifos 94 factory effluent.
Summary of the invention
Goal of the invention: in order to overcome the deficiencies in the prior art, the first object of the present invention is to provide a kind of Pseudomonas aeruginosa Chlorpyrifos 94 to Degradation.
The second object of the present invention is to provide a kind of application be prepared into by described Pseudomonas aeruginosa after immobilized spherule in chlorpyrifos degrading.
Technical scheme: in order to solve above-mentioned first object, the technical solution adopted in the present invention is: a kind of Pseudomonas aeruginosa, its Classification And Nomenclature is Pseudomonas aeruginosa (Pseudomonasaeruginosa) DSP-05, be deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center (being called for short CGMCC), depositary institution address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode is 100101, deposit number is: CGMCCNo.8390, and preservation date is on October 24th, 2013.
In order to solve the second above-mentioned object, the technical solution adopted in the present invention for: described Pseudomonas aeruginosa be prepared into immobilized spherule after bacillus amyloliquefaciens described in application in chlorpyrifos degrading suppressing the application in the growth of strawberry anthrax-bacilus.
Have particular application as: first activated by immobilized spherule aeration 12h in Chlorpyrifos 94 factory effluent, water temperature 26 DEG C-30 DEG C in reactivation process, aeration rate is 1.2L/min; Then temperature be 26 DEG C-30 DEG C, under pH value is 6.8-7.2, Chlorpyrifos 94 influent concentration 200mg/L, hydraulic detention time be the condition of 32h, often liter of Chlorpyrifos 94 waste water is taked the immobilized spherule of 185mL to process.
Further, the method described Pseudomonas aeruginosa being prepared into immobilized spherule comprises the following steps:
(1) use a small amount of Pseudomonas aeruginosa strain inoculation of transfering loop picking on slant medium in 30 DEG C in LB liquid nutrient medium, under 160r/min condition, shaking culture is to logarithmic phase;
(2) by above-mentioned cultured bacterium liquid centrifugal 2min under the condition of 8000r/min, outwell supernatant liquor, add the sterilized water of same volume, after shaking up under the condition of 8000r/min centrifugal 2min, repetitive operation twice, suspends with the sterilized water of same volume, makes thalline liquid;
(3) mass concentration of getting 1mL thalline liquid and 20mL be 4% sodium alginate soln mix in room temperature, then with syringe, by mixed solution, the mass concentration instilled under room temperature is the CaCl of 3%
2solution, obtains calcium alginate gel immobilized spherule after crosslinked calcification 6h at 4 DEG C, aseptic washing be stored in for twice 4 DEG C for subsequent use.
Further, in above-mentioned steps (1), the formula of LB substratum is: 1000mL distilled water, yeast powder 5.0g, peptone 10.0g, sodium-chlor 10.0g, pH7.0.
The physiologically active feature of Pseudomonas aeruginosa (Pseudomonasaeruginosa) DSP-05 described in the present invention is as follows:
Described Pseudomonas aeruginosa (Pseudomonasaeruginosa) DSP-05 Main Biological is G-, this bacterium oxidase positive, and energy oxygenolysis glucose and xylose, produces acid not aerogenesis, but do not reduce lactose and sucrose.Liquefy gelatin, decomposable asymmetric choice net urea, reduction nitrate is nitrite and produces nitrogen.
Gramstaining shows as feminine gender; Indoles is negative, utilizes citrate, and arginine dihydrolase is positive; By the 16SrDNA gene order of this bacterial strain of BLAST comparison, identify that it is Pseudomonas aeruginosa (Pseudomonasaeruginosa).
Beneficial effect: compared with prior art, advantage of the present invention is:
(1) a kind of Pseudomonas aeruginosa (Pseudomonasaeruginosa) DSP-05 disclosed by the invention can be that sole carbon source grows with Chlorpyrifos 94; Under laboratory shake flask condition, 93% is reached to the degradation rate of 200mg/L Chlorpyrifos 94.Present invention also offers a kind of immobilized spherule used prepared by bacterial strain DSP-05, laboratory biological degradation experiment result shows, it reaches 90% to the degradation rate of 200mg/L Chlorpyrifos 94.It is low, easy to use that the preparation method of immobilized spherule has production cost, the advantage that removal effect is good, can Chlorpyrifos 94 in degrading waste water, is adapted at promoting the use of in the wastewater treatment containing Chlorpyrifos 94.The present invention is for preserving the ecological environment, and the protection mankind's is healthy, reduces cost for wastewater treatment and has great importance.
(2) Pseudomonas aeruginosa (Pseudomonasaeruginosa) DSP-05 culture condition simply, is easily preserved, and is easy to suitability for industrialized production, has good development prospect.
Accompanying drawing explanation
Fig. 1 is bacterial strain DSP-05 thalline violet staining photo (1000x) of the present invention;
Fig. 2 be the present invention use bacterial strain DSP-05 to prepare immobilized spherule to the time dependent schematic diagram of chlorpyrifos degrading clearance;
Fig. 3 is the degraded clearance schematic diagram of the present invention's immobilized spherule of using bacterial strain DSP-05 to prepare to the different influent concentration of Chlorpyrifos 94.
Embodiment
The application of Pseudomonas aeruginosa of the present invention (Pseudomonasaeruginosa) DSP-05 is elaborated below in conjunction with experimental example.
The separation andpreconcentration method of Pseudomonas aeruginosa (Pseudomonasaeruginosa) DSP-05 bacterial strain:
(1) be separated: get Chlorpyrifos 94 enrichment bacterium liquid 1.0ml, add in 9.0mL sterilized water, fully mixing is made into 10
-1pregnant solution, then draw that 1.0mL prepares 10
-1pregnant solution add in 9.0mL sterilized water, fully mixing is made into 10
-2pregnant solution, by that analogy, gradient dilution is carried out to pregnant solution.The diluent 0.1mL drawing each gradient coats on the inorganic salt solid medium containing 200mg/L Chlorpyrifos 94 that (formula of inorganic salt solid medium is: 1000mL distilled water, 1.5gK
2hPO
4, 0.5gKH
2pO
4, 0.2gMgSO
47H
2o, 1.0gNaCl, 1.0g (NH
4)
2sO
4, 20g agar, pH7.0), cultivate 7 days for 30 DEG C; From picking list bacterium colony above inorganic salt solid medium after 7 days, cultivate in the LB liquid nutrient medium of 5mL after 24 hours, centrifugal 2min under the condition of 8000r/min, remove supernatant liquor, the sterilized water adding 5mL shakes up, still centrifugal 2min under the condition of 8000r/min, according to said method with after aseptic washing twice, adds 5mL sterilized water and to suspend this bacterium; This bacterium liquid of absorption 1mL adds the inorganic salt liquid substratum that 100mL Chlorpyrifos 94 concentration is 200mg/L, and (formula is: 1000mL distilled water, 1.5gK
2hPO
4, 0.5gKH
2pO
4, 0.2gMgSO
47H
2o, 1.0gNaCl, 1.0g (NH
4)
2sO
4, pH7.0) and in 160r/min, under 30 DEG C of conditions, shaking culture is after 3 days, surveys its degradation effect by ultraviolet spectrophotometry.As shown in Figure 2, under laboratory shake flask condition, the degradation rate of free bacteria to 200mg/L Chlorpyrifos 94 reaches 90%.
(2) identify: a strain bacterial strain higher for degradation efficiency is preserved, carries out subsequent experimental.Its thalline gramstaining photo (1000x) as shown in Figure 1, is accredited as P. aeruginosa Pseudomonas (Pseudomonasaeruginosa.p.); Called after: DSP-05.Main Biological is G
-, logarithmic phase thalline is elongated and different in size, sometimes in club shape or wire, and paired or short catenation.This bacterium oxidase positive, energy oxygenolysis glucose and xylose, produces acid not aerogenesis, but does not reduce lactose and sucrose.Liquefy gelatin, decomposable asymmetric choice net urea, reduction nitrate is nitrite and produces nitrogen, and indoles is negative, utilizes citrate, and arginine dihydrolase is positive.Can be that sole carbon source grows with Chlorpyrifos 94.
Pseudomonas aeruginosa is prepared into the method for immobilized spherule:
(1) use a small amount of Pseudomonas aeruginosa strain inoculation of transfering loop picking on slant medium in 30 DEG C in LB liquid nutrient medium, under 160r/min condition, shaking culture is to logarithmic phase;
(2) by above-mentioned cultured bacterium liquid centrifugal 2min under the condition of 8000r/min, outwell supernatant liquor, add the sterilized water of same volume, after shaking up under the condition of 8000r/min centrifugal 2min, repetitive operation twice, suspends with the sterilized water of same volume, makes thalline liquid;
(3) mass concentration of getting 1mL thalline liquid and 20mL be 4% sodium alginate soln mix in room temperature, then with syringe, by mixed solution, the mass concentration instilled under room temperature is the CaCl of 3%
2solution, obtains calcium alginate gel immobilized spherule after crosslinked calcification 6h at 4 DEG C, aseptic washing be stored in for twice 4 DEG C for subsequent use.
Wherein in above-mentioned steps (1), the formula of LB substratum is: 1000mL distilled water, yeast powder 5.0g, peptone 10.0g, sodium-chlor 10.0g, pH7.0.
Embodiment 1:
Immobilized spherule is applied to the method for chlorpyrifos degrading: first activated by immobilized spherule aeration 12h in Chlorpyrifos 94 factory effluent, water temperature 26 DEG C in reactivation process, aeration rate is 1.2L/min; Then temperature be 26 DEG C, under pH value is 6.8, Chlorpyrifos 94 influent concentration 200mg/L, hydraulic detention time be the condition of 32h, often liter of Chlorpyrifos 94 waste water is taked the immobilized spherule of 185mL to process.
Embodiment 2:
First activated by immobilized spherule aeration 12h in Chlorpyrifos 94 factory effluent, water temperature 30 DEG C in reactivation process, aeration rate is 1.2L/min; Then temperature be 30 DEG C, under pH value is 7.2, Chlorpyrifos 94 influent concentration 200mg/L, hydraulic detention time be the condition of 32h, often liter of Chlorpyrifos 94 waste water is taked the immobilized spherule of 185mL to process.
Embodiment 3:
First activated by immobilized spherule aeration 12h in Chlorpyrifos 94 factory effluent, water temperature 28 DEG C in reactivation process, aeration rate is 1.2L/min; Then temperature be 28 DEG C, under pH value is 7.0, Chlorpyrifos 94 influent concentration 200mg/L, hydraulic detention time be the condition of 32h, often liter of Chlorpyrifos 94 waste water is taked the immobilized spherule of 185mL to process.
Adopt the method for above-described embodiment 1-3 for Chlorpyrifos 94 removal situation as shown in Figure 2, the clearance of Chlorpyrifos 94, more than 90%, has good removal effect.
As shown in Figure 3, for the removal situation of the Chlorpyrifos 94 of different influent concentration, research shows, the clearance of Chlorpyrifos 94 reduces along with the rising of influent concentration, when intake Chlorpyrifos 94 concentration lower than in the scope of 500mg/L time, Chlorpyrifos 94 has higher clearance, can reach more than 85%.
Claims (6)
1. a Pseudomonas aeruginosa, its Classification And Nomenclature is Pseudomonas aeruginosa (Pseudomonasaeruginosa) DSP-05, be deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is: CGMCCNo.8390, and preservation date is on October 24th, 2013.
2. the application of Pseudomonas aeruginosa according to claim 1 in chlorpyrifos degrading.
3. application according to claim 2, is characterized in that, described Pseudomonas aeruginosa is prepared into the application in chlorpyrifos degrading after immobilized spherule.
4. application according to claim 3, is characterized in that, is first activated by immobilized spherule aeration 12h in Chlorpyrifos 94 factory effluent, water temperature 26 DEG C-30 DEG C in reactivation process, and aeration rate is 1.2L/min; Then temperature be 26 DEG C-30 DEG C, under pH value is 6.8-7.2, Chlorpyrifos 94 influent concentration 200mg/L, hydraulic detention time be the condition of 32h, often liter of Chlorpyrifos 94 waste water is taked the immobilized spherule of 185mL to process.
5. application according to claim 3, is characterized in that, the method that described Pseudomonas aeruginosa is prepared into immobilized spherule is comprised the following steps:
(1) use a small amount of Pseudomonas aeruginosa strain inoculation of transfering loop picking on slant medium in 30 DEG C in LB liquid nutrient medium, under 160r/min condition, shaking culture is to logarithmic phase;
(2) by above-mentioned cultured bacterium liquid centrifugal 2min under the condition of 8000r/min, outwell supernatant liquor, add the sterilized water of same volume, after shaking up under the condition of 8000r/min centrifugal 2min, repetitive operation twice, suspends with the sterilized water of same volume, makes thalline liquid;
(3) mass concentration of getting 1mL thalline liquid and 20mL be 4% sodium alginate soln mix in room temperature, then with syringe, by mixed solution, the mass concentration instilled under room temperature is the CaCl of 3%
2solution, obtains calcium alginate gel immobilized spherule after crosslinked calcification 6h at 4 DEG C, aseptic washing be stored in for twice 4 DEG C for subsequent use.
6. application according to claim 5, is characterized in that: in described step (1), the formula of LB substratum is: 1000mL distilled water, yeast powder 5.0g, peptone 10.0g, sodium-chlor 10.0g, pH7.0.
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| CN104129852B (en) * | 2014-06-20 | 2016-05-11 | 浙江工商大学 | A kind of refuse landfill H2The method of the endogenous reduction of S and reaction unit |
| CN104250629A (en) * | 2014-07-09 | 2014-12-31 | 浙江师范大学 | Pseudomonas fluorescens CHZYR6 3 strain and enzyme preparation thereof and application thereof to degradation of chlopyrifos pesticide |
| CN105886426B (en) * | 2016-03-18 | 2019-11-15 | 天津大学 | Oil Spill Restoration Bacteria and Its Application in Polluted Environment |
| CN106754513B (en) * | 2016-12-22 | 2020-08-14 | 浙江工商大学 | Preparation and Application of TX-100 Modified Sodium Alginate-Embedded Pseudomonas Particles |
| CN113897407B (en) * | 2021-10-27 | 2023-10-24 | 长春工业大学 | Pseudomonas aeruginosa and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101096644A (en) * | 2006-06-26 | 2008-01-02 | 谢明 | Pseudomonas stutzeri JSD-008 and its degradation function for organophosphorus pesticide |
| CN102533602A (en) * | 2012-01-05 | 2012-07-04 | 陕西延长石油(集团)有限责任公司研究院 | Pseudomonas aeruginosa, and culture method and application thereof |
| CN103114066A (en) * | 2013-03-15 | 2013-05-22 | 江苏南资环保科技有限公司 | Diethylamine degradation pseudomonas strain EYA-2 and application thereof |
| CN103468597A (en) * | 2012-10-11 | 2013-12-25 | 广西科学院 | Pseudomonas aeruginosa producing alkaline cellulases |
| CN103525802A (en) * | 2013-10-24 | 2014-01-22 | 东华大学 | Preparation method of immobilized globules of klebsiella pneumoniae |
-
2014
- 2014-03-07 CN CN201410083918.7A patent/CN103820370B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101096644A (en) * | 2006-06-26 | 2008-01-02 | 谢明 | Pseudomonas stutzeri JSD-008 and its degradation function for organophosphorus pesticide |
| CN102533602A (en) * | 2012-01-05 | 2012-07-04 | 陕西延长石油(集团)有限责任公司研究院 | Pseudomonas aeruginosa, and culture method and application thereof |
| CN103468597A (en) * | 2012-10-11 | 2013-12-25 | 广西科学院 | Pseudomonas aeruginosa producing alkaline cellulases |
| CN103114066A (en) * | 2013-03-15 | 2013-05-22 | 江苏南资环保科技有限公司 | Diethylamine degradation pseudomonas strain EYA-2 and application thereof |
| CN103525802A (en) * | 2013-10-24 | 2014-01-22 | 东华大学 | Preparation method of immobilized globules of klebsiella pneumoniae |
Non-Patent Citations (1)
| Title |
|---|
| "铜绿假单胞菌对DBP的降解特性研究";吴为中 等;《环境科学》;20090228;第30卷(第2期);第510页第1.1节 * |
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