CN107446840A - One plant of 2 pyridazinone degradation bacteria of 6 hydroxyl of phenyl 3 and its application - Google Patents
One plant of 2 pyridazinone degradation bacteria of 6 hydroxyl of phenyl 3 and its application Download PDFInfo
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Abstract
The invention provides a kind of degradation bacteria of the pyridazinone of 2 phenyl, 6 hydroxyl 3, the degrading strain identification is aerosporusBacillus aerophilus, belong to Gram-positive mushroom, it is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on May 26th, 2017（CGMCC）, deposit number：CGMCC NO.14198; it can be grown using the pyridazinone of 2 phenyl, 6 hydroxyl 3 as sole carbon source and the energy; to the degradation rate of the pyridazinone of 100 mg/L, 2 phenyl, 6 hydroxyl 3 up to more than 90% in 32 h; the pyridazinone difficult degradation of 2 phenyl, 6 hydroxyl 3 caused by solving the problems, such as during pesticide producing; pesticide wastewater qualified discharge can be made, important function is served to the clean manufacturing and environmental protection of the pyridazinone of 2 phenyl, 6 hydroxyl 3.
Description
Technical field
The invention belongs to biological modeling technical field, and in particular to a kind of 2- phenyl -6- hydroxyls -3- pyridazinone degradation bacterias
And its application.
Background technology
Organophosphorus pesticide has made significant contribution in agricultural production, and 2- phenyl -6- hydroxyl -3- pyridazinones are organophosphorus insecticidals
The intermediate of worm agent pyridaphethione, because it has unique heterocycle structure, therefore in process of production, caused waste water is so far
Do not administered effectively also, also bring serious problem of environmental pollution, it pollutes the reparation of environment and causes extensive pass
Note.
Microbial degradation has the advantages that low cost, efficiency high, non-secondary pollution, ecological recovery are good, in many sides
Face is applied, so filtering out 2- phenyl -6- hydroxyls -3- pyridazinones degradation bacteria for handling 2- phenyl -6- hydroxyl -3- pyridazines
Ketone waste water is significant.
The content of the invention
It is an object of the invention to provide a kind of 2- phenyl -6- hydroxyls -3- pyridazinones degradation bacteria and its application, this bacterium is used
Obtained microbial inoculum can make the degradation rate of 2- phenyl -6- hydroxyl -3- pyridazinones, and up to more than 90%, production and use cost are relatively low.
The present invention provides a kind of 2- phenyl -6- hydroxyls -3- pyridazinone degradation bacterias, and its bacterial strain NZJ-52 is one plant of gram sun
Property bacterium, Main Biological be thalline it is shaft-like, bacterium colony is white, opaque, and edge is imperfect, central spore, diameter 1-2
Mm, optimum growth temperature are 30 DEG C, pH 7.0.It is preserved in Chinese microorganism strain preservation management on May 27th, 2017
Committee's common micro-organisms center（CGMCC）, address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, deposit number：CGMCC
NO.14198, identified is aerosporusBacillus aerophilus, can using 2- phenyl -6- hydroxyl -3- pyridazinones as
Sole carbon source, nitrogen source are grown, and its mineralising is discharged into ammonia nitrogen.
The invention provides the microbial inoculum of the aerosporus NZJ-52.
Present invention also offers the method for aerosporus NZJ-52 production microbial inoculums.
The packaging formulation of the product is liquid bacterial agent or solid absorption microbial inoculum.
The method of the production microbial inoculum, is concretely comprised the following steps：
1）2- phenyl -6- hydroxyl -3- pyridazinone degradation bacteria aerosporus NZJ-52 test tube kind is inoculated in LB culture mediums
In, shaken cultivation to logarithmic phase, LB culture medium prescriptions are：The g/L of NaCl 10.00, the g/L of peptone 10.00, dusty yeast 5.00
G/L, pH 7.0；
2）By step 1）Cultured strain is inoculated with into 500L seeding tanks by 10% inoculum concentration, is cultivated to exponential phase, kind
Culture medium prescription used in sub- tank is：Glucose 0.8%,（NH4）2SO4 1%, K2HPO4 0.2%, MgSO40.05%, NaCl
0.01%, CaCO30.3%, yeast extract 0.02%, pH value 7.2-7.5.
3）By step 2）The seed liquor carries out fermented and cultured by 10% inoculum concentration access production tank, produces used in tank
Culture medium is identical with seed tank culture base, and the nutrient solution after the completion of fermentation is microbial inoculum.
Step 2）The seeding tank and step 3）In the incubation of described production tank, the throughput of filtrated air is 1：
0.6-1.2, mixing speed are 180-240 r/min, and cultivation temperature is 30-35 DEG C, step 2 and 3 whole technological process culture
Time is 48-60 h.
Application of the microbial inoculum in the production waste water of processing organophosphorus insecticide pyridaphethione, degraded object is 2- benzene
Base -6- hydroxyl -3- pyridazinones.
Beneficial effect：
Aerosporus NZJ-52 of the present invention to the degradation rates of 2- phenyl -6- hydroxyl -3- pyridazinones up to more than 90%,
The workload during production and use is greatly reduced, reduces production and use cost.The present invention is for protecting ecology ring
Border, the cost for reducing wastewater treatment have great importance.Prepared microbial inoculum has that production and application cost is low, easy to use,
The advantages of removal effect is good.Degradation bacteria NZJ-52 can be in the culture medium using 2- phenyl -6- hydroxyl -3- pyridazinones as sole carbon source
Upper growth；Degradation bacteria NZJ- is accessed in the minimal medium of -6- hydroxyl -3- pyridazinones of phenyl containing 2- by 1% inoculum concentration
52, under the conditions of 30 DEG C, pH 7.0, the degraded in the h of shaken cultivation 32 to 100 mg/L 2- phenyl -6- hydroxyl -3- pyridazinones
Rate solves the problems, such as caused 2- phenyl -6- hydroxyls -3- pyridazinone difficult degradations during pesticide producing, can made up to more than 90%
Pesticide wastewater qualified discharge, important work is served to the clean manufacturing and environmental protection of 2- phenyl -6- hydroxyl -3- pyridazinones
With.
Brief description of the drawings
Fig. 1 aerosporus NZJ-52 colonial morphology figure；
Fig. 2 aerosporus NZJ-52 shaking flask degraded figure.
Embodiment
The separation and identification of the bacterial strain of embodiment 1
2- phenyl -6- hydroxyl -3- pyridazinones are made into 10-1Pregnant solution, then draw 1.0 ml are prepared 10-1Pregnant solution add
Enter in 9.0 ml sterilized waters, fully mix and be made into 10-2Pregnant solution, by that analogy, to pregnant solution carry out gradient dilution.Draw each
The concentration that the ml of dilution 0.1 of gradient is coated on the -6- hydroxyl -3- pyridazinones of phenyl containing 2- is trained for 100 mg/L inorganic salts solids
Supporting base, (formula is：Every liter contains 1.50 g K2HPO4、0.50 g KH2PO4、0.20 g MgSO4×7H2O、1.00 g NaCl、
1.00 g (NH4)2SO4, 20.00 g agar, pH 7.0) on, 30 DEG C are cultivated 7 days.Therefrom picking single bacterium colony verifies its degraded
Effect, one plant of higher bacterial strain of degradation efficiency is preserved, carries out subsequent experimental.Bacterium colony figure is as shown in figure 1, be accredited as thermophilic gas gemma
BacillusBacillus aerophilus.Main Biological is that thalline is shaft-like, and bacterium colony is white, opaque, and edge is endless
Whole, central spore, diameter 1-2mm, optimum growth temperature is 30 DEG C, pH 7.0.Can using 2- phenyl -6- hydroxyl -3- pyridazinones as
Sole carbon source, nitrogen source are grown, and its mineralising is produced into ammonia nitrogen.
The aerosporus NZJ-52 of embodiment 2 shaking flask degradation experiment
In the minimal medium of the 2- phenyl -6- hydroxyl -3- pyridazinones containing 100mg/L, its formula is：Every liter contains 1.50 g
K2HPO4,0.50g KH2PO4,0.20g MgSO4 × 7H2O, 1.00g NaCl, 1.00g (NH4) 2SO4, pH 7.0, by 1%
Inoculum concentration inoculation NZJ-52, in 30 DEG C of shaken cultivations, be measured by sampling every 4 h.As seen from Figure 2,2- phenyl -6- in 32 h
Hydroxyl -3- pyridazinones are degraded more than 90%.
Preparation of the embodiment 3 containing aerosporus NZJ-52 microbial inoculums
Aerosporus NZJ-52 is activated on culture dish, is inoculated in test tube slant（Culture medium prescription：NaCl 10.00
G/L, the g/L of peptone 10.00, the g/L of dusty yeast 5.00,2% agar, pH 7.0）It is upper standby.Test tube kind is inoculated in containing 200ml
In the 1000ml shaking flasks of LB culture mediums, LB culture medium prescriptions are：Yeast extract 5.00g/L, peptone 10.00g/L, NaCl
10.00g/L, pH 7.0, constant-temperature shaking culture to logarithmic phase, prepare inoculation seeding tank.500 liters of seeding tank, 400 liters of inventory,
Culture medium prescription is：Glucose 0.8%,（NH4）2SO4 1%, K2HPO4 0.2%, MgSO40.05%, NaCl 0.01%, CaCO3
0.3%, yeast extract 0.02%, pH value 7.2-7.5.121 DEG C of high pressure moist heat sterilizations after feeding intake, will be above-mentioned after being cooled to 33 DEG C
Cultured shaking flask strain is inoculated with into 500 liters of seeding tanks by 10% inoculum concentration, is cultivated to exponential phase, mixing speed and is
220 revs/min, filtrated air intake is 1:0.8.The seed liquor for reaching logarithmic phase is trained by 10% inoculum concentration access production tank
Support, production tank used medium composition is identical with seed tank culture base.Produce 5 tons of tankage size, 4.5 tons of inventory.After feeding intake
Produce tank 1.1kg/cm2Pressure under, 121 DEG C of high pressure moist heat sterilizations, less than 35 DEG C are cooled to after sterilizing, lead to filtrated air keep
Germ-free condition is standby.Production tank temperature control after inoculation produces the throughput of filtrated air in the incubation of tank at 35 DEG C
For 1:1.2, mixing speed is 240 revs/min, and whole technological process incubation time is 60 hours.Thalline quantity reaches after fermentation ends
To 1,000,000,000/more than ml.After the completion of fermentation nutrient solution go out tank directly with plastic barrel or Packaging Bottle be distributed into liquid dosage form or
Solid fungicide formulation is distributed into packaging bag using adsorption by peat.
The aerosporus NZJ-52 of embodiment 4 answering in the production waste water of processing organophosphorus insecticide pyridaphethione
With
Certain 1 ton of organophosphorus pesticide factory waste water is taken, by coagulation, consolidating in waste water is removed with grid, screen cloth, sedimentation basin
Body suspension, the mg/L of 2- phenyl -6- hydroxyl -3- pyridazinones content 5820 in initial waste, it is preliminary adjust pH value after put into 10
Kg solid fungicides, it is 232.8 mg/L that 2- phenyl -6- hydroxyl -3- pyridazinones contents are determined after 32 h, finds the drop to benzene azoles alcohol
Solution rate reaches 96%.
Claims (7)
1. one plant of 2- phenyl -6- hydroxyl -3- pyridazinone degradation bacteria, its Classification And Nomenclature is aerosporus（Bacillus aerophilus）NZJ-52, has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and preservation is compiled
Number：CGMCC NO.14198.
2. include a kind of microbial inoculum of 2- phenyl -6- hydroxyls -3- pyridazinone degradation bacterias described in claim 1.
3. the preparation method of microbial inoculum described in claim 2, it is characterised in that comprise the following steps：
Step 1）, the test tube kind of 2- phenyl -6- hydroxyl -3- pyridazinone degradation bacterias is inoculated in LB culture mediums, shaken cultivation is extremely
Logarithmic phase；
Step 2）, above-mentioned cultured strain is inoculated with into seeding tank by 10% inoculum concentration, cultivated to exponential phase；
Step 3）, by seed liquor by 10% inoculum concentration access production tank culture, produce tank used medium and seed tank culture base
It is identical；
The throughput of filtrated air is 1 in the incubation of seeding tank and production tank：0.6-1.2, mixing speed 180-240
R/min, cultivation temperature are 30-35 DEG C, and whole technological process incubation time is 48-60 h, and thalline quantity reaches after fermentation ends
1000000000/more than mL, nutrient solution goes out tank direct packaging into liquid bacterial agent after the completion of fermentation, or using adsorption by peat packaging bag point
Dress up solid fungicide.
4. the preparation method of microbial inoculum according to claim 3, it is characterised in that the LB culture medium prescriptions are：NaCl
10.00 g/L, the g/L of peptone 10.00, dusty yeast 5.00 g/L, pH 7.0.
5. the preparation method of microbial inoculum according to claim 3, it is characterised in that the culture medium prescription used in the seeding tank
For：Glucose 0.8%,（NH4）2SO4 1%, K2HPO4 0.2%, MgSO40.05%, NaCl 0.01%, CaCO30.3%, yeast extract
0.02%, pH value 7.2-7.5.
6. 2- phenyl -6- hydroxyls -3- pyridazinones degradation bacteria described in claim 1 is in the life of processing organophosphorus insecticide pyridaphethione
Produce the application in waste water.
7. microbial inoculum described in 2- phenyl -6- hydroxyls -3- pyridazinones degradation bacteria described in claim 1 or claim 2 is in degraded 2-
Application in phenyl -6- hydroxyl -3- pyridazinones.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1302865A (en) * | 2001-01-17 | 2001-07-11 | 南京农业大学 | Bacterium for degradating residual agricultural organophosphorus chemical and its bacterial preparation |
| CN103952353A (en) * | 2014-04-24 | 2014-07-30 | 烟台海上传奇生物科技有限公司 | Bacillus aerophilus, microbial agent and applications of bacillus aerophilus and microbial agent |
-
2017
- 2017-07-10 CN CN201710556797.7A patent/CN107446840B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1302865A (en) * | 2001-01-17 | 2001-07-11 | 南京农业大学 | Bacterium for degradating residual agricultural organophosphorus chemical and its bacterial preparation |
| CN103952353A (en) * | 2014-04-24 | 2014-07-30 | 烟台海上传奇生物科技有限公司 | Bacillus aerophilus, microbial agent and applications of bacillus aerophilus and microbial agent |
Non-Patent Citations (2)
| Title |
|---|
| ROMILA AKOIJAM ET AL.: "Biodegradation of imidacloprid in sandy loam soil by Bacillus aerophilus", 《INTERN. J. ENVIRON. ANAL. CHEM.》 * |
| 李荣等: "菌株 scl-2 对苯线磷、哒嗪硫磷和丙溴磷降解的研究", 《中国环境科学》 * |
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