CN106085899B - Microbial inoculum and the application of one plant of process for preparation of benzoylurea compounds degradation bacteria and its production - Google Patents
Microbial inoculum and the application of one plant of process for preparation of benzoylurea compounds degradation bacteria and its production Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/025—Achromobacter
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D3/00—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances
- A62D3/02—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances by biological methods, i.e. processes using enzymes or microorganisms
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D2101/00—Harmful chemical substances made harmless, or less harmful, by effecting chemical change
- A62D2101/04—Pesticides, e.g. insecticides, herbicides, fungicides or nematocides
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- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D2101/00—Harmful chemical substances made harmless, or less harmful, by effecting chemical change
- A62D2101/20—Organic substances
- A62D2101/26—Organic substances containing nitrogen or phosphorus
Abstract
The invention discloses one plant of process for preparation of benzoylurea compounds degradation bacteria strains M6, it is identified as achromobacter (Achromobacter dolens), it is preserved in China typical culture collection center, the preservation time is on May 11st, 2016, and deposit number is CCTCC NO:M 2016257；The invention also discloses application of the degradation bacteria strains M6 in production process for preparation of benzoylurea compounds degradation bacterial agent, the degradation bacterial agent can degrade a variety of process for preparation of benzoylurea compounds such as flubenzuron, diflubenzuron, d ichlorbenzuron, can make in a short time remaining flubenzuron in soil or water body environment, diflubenzuron, d ichlorbenzuron degradation rate up to 90% or more；Degradation bacterial agent of the invention can be produced with the general Zymolysis Equipment of fermentation industry, have production cost low, easy to use, the good advantage of removal effect.
Description
Technical field
The present invention relates to environmental pollution microorganism remediations, and in particular to one plant of process for preparation of benzoylurea compounds degradation bacteria and its life
The microbial inoculum of production and application.
Background technique
Process for preparation of benzoylurea compounds, including flubenzuron, diflubenzuron, d ichlorbenzuron etc. are chitin desinsection inhibitor, pass through suppression
Chitin synthesis processed causes insect death.Process for preparation of benzoylurea compounds shows very abnormal insect, especially lepidopterous larvae
Good insecticidal activity, for preventing and treating various pests, such as mythimna separata, beet armyworm, prodenia litura, poison moth, cabbage caterpillar, pine moth, small
Diamond-back moth, rice leaf roller, oriental fruit months etc., and the pest of resistance is generated to organic phosphorus, carbamates, pyrethroid
With good result, can also prevent and treat Brontispa longissima with the co-administrations such as avermectin, be applied to extensively rice, pakchoi, rape,
The crops such as lichee, longan, tea.Global process for preparation of benzoylurea compounds sales volume is 7.19 hundred million dollars within 2014, and is in sustainable growth
Trend.
Process for preparation of benzoylurea compounds drug effect speed is slower, and the residual period is longer, and principal item diflubenzuron is in different type farmland
Half life of decomposition in soil is 8~16 weeks.Such pesticide main environment behavior has leaching, hydrolysis, photodissociation, microbial degradation.
Remains of pesticide focuses primarily upon in topsoil, is slowly migrated to deep layer due to eluviation；Benzoyl area kind in soil
Insecticide moves to neighbouring river with runoff, in lake, due to sunlight irradiation and photodissociation occurs, the pesticide in atmosphere
Understand the sedimentation due to particulate matter and enter water body, water body is made to become another important place existing for such pesticide；In water body and
Hydrolysis and microbial degradation can all occur in soil, and microorganism plays a leading role in process for preparation of benzoylurea compounds degradation.
After process for preparation of benzoylurea compounds application, phytotoxicity will be caused in the case where strong light, high temperature, high concentration；Such pesticide
The residual period is longer in the soil, and decomposition product also has larger toxicity, and residual mixture on vegetables leads to mouse DNA double-strand
Fracture, generate carcinogenic, teratogenic effect or other serious diseases；And itself and its decomposition product can enter ground from soil mobility
It is lauched, causes production and living pollution of waterhead；To silkworm, honeybee and aquatic shell-fish and arthropods animal, especially to its young
Murder by poisoning it is larger, have genetoxic.
Less to process for preparation of benzoylurea compounds degradation bacteria report both at home and abroad, degradation bacteria type only has Penicillium notatum, bud branch at present
Mould, Aspergillus, Trichoderma, Rhodococcus sp, secondary coccus, degradation efficiency 67%~89% differ.But it up to the present, yet there are no energy
The research for a variety of process for preparation of benzoylurea compounds bioremediation technology patents such as flubenzuron, diflubenzuron, d ichlorbenzuron of degrading and application report
Road.
Summary of the invention
Goal of the invention: in view of the problems of the existing technology, the present invention provides one plant of process for preparation of benzoylurea compounds degradation bacteria,
For remaining flubenzuron, diflubenzuron, d ichlorbenzuron process for preparation of benzoylurea compounds in degrade soil or water body environment；Of the invention is another
One purpose is to provide the microbial inoculum of process for preparation of benzoylurea compounds degradation bacteria production；The degradation bacterial agent of bacterial strain preparation can degrade fluorine
A variety of process for preparation of benzoylurea compounds such as bell urea, diflubenzuron, d ichlorbenzuron, this microbial inoculum can make in a short time in soil or water body environment
Remaining flubenzuron, diflubenzuron, d ichlorbenzuron degradation rate up to 90% or more；Degradation bacterial agent of the invention can be logical with fermentation industry
It is produced with Zymolysis Equipment, has production cost low, easy to use, the good advantage of removal effect.
Technical solution: to achieve the goals above, one plant of process for preparation of benzoylurea compounds degradation bacteria strains M6 as described in invention,
It is identified as achromobacter (Achromobacter dolens), has been preserved in China typical culture collection center (referred to as
CCTCC), preservation address: the Chinese Wuhan Wuhan University, postcode: 430072, the preservation time is on May 11st, 2016, and preservation is compiled
Number be CCTCC NO:M 2016257.The bacterial strain is inventor from Haikou city Hainan Run He pesticide Co., Ltd agricultural chemicals waste water
It is separated in the activated sludge of biochemical treatment tank.Degradation bacteria strains M6 Main Biological are as follows: G?, raw on solid medium
After long 3d, bacterium colony dimpling, translucent, non-pigment, by negative staining, the bacterium is in the shape of a rod under transmission electron microscope, amphitrichous, nothing
Pod membrane does not form gemma, and size is 0.4-0.7 × 1.4-2.8 μm, and aerobic growth, nitrate reaction is the positive, using grape
Sugar, gluconic acid, adipic acid, citric acid have alkaline phosphatase activities, and nitrite, indoles generate, gelatin liquefaction reaction is negative
Property.
Application of the degradation bacteria strains M6 of the present invention in degradation process for preparation of benzoylurea compounds.
Further, the degradation bacteria strains M6 answering in process for preparation of benzoylurea compounds in degradation soil or water body environment
With.
Further, the process for preparation of benzoylurea compounds is one of flubenzuron, diflubenzuron, d ichlorbenzuron or a variety of.
Process for preparation of benzoylurea compounds flubenzuron, diflubenzuron, d ichlorbenzuron in the present invention are bought in the brilliant pure biochemistry in Aladdin reagent Shanghai
Limited liability company.
Application of the degradation bacteria strains M6 of the present invention in production process for preparation of benzoylurea compounds bacterium for degradating residual agricultural chemical.
The present invention also provides a kind of degradation bacterial agents produced using the bacterial strain M6.
Further, the degradation bacterial agent, by made by following steps:
1) the cultured test tube liquid of degradation bacteria strains M6 is inoculated in fermentation medium by the inoculum concentration of 1-1.5% volume ratio
In, fermenting microbe is made in shaken cultivation to logarithmic phase；
2) above-mentioned cultured fermenting microbe is inoculated in the culture medium of seeding tank by the inoculum concentration of 8-10% volume ratio,
Seed liquor is made to logarithmic growth phase in culture；
3) it is cultivated in the culture medium by seed liquor by the inoculum concentration access production tank of 8-10% volume ratio；
4) ventilatory capacity of filtrated air is 1:(0.8-1.0 in the incubation of seeding tank and production tank), mixing speed
For 170-230rpm, cultivation temperature is 28-32 DEG C, and whole process incubation time is 36-48 hours, and thalline quantity reaches after fermentation
To 1,000,000,000/mL or more, culture solution goes out tank and is directly distributed into liquid agent with plastic barrel or Packaging Bottle after fermentation.
Further, the degradation bacterial agent, wherein the training of the culture medium of the fermentation medium, seeding tank, production tank
It is identical to support based formulas, is glucose 0.1wt%, NaCl1.0wt%, peptone 0.5wt%, yeast extract 0.25wt%,
pH7.2-7.5。
Application of the degradation bacterial agent of the present invention in degradation process for preparation of benzoylurea compounds.
Further, the degradation bacterial agent answering in process for preparation of benzoylurea compounds in degradation soil or water body environment
With.
Further, the process for preparation of benzoylurea compounds is one of flubenzuron, diflubenzuron, d ichlorbenzuron or a variety of.
The utility model has the advantages that being compared by the prior art, the present invention obtains one plant of achromobacter through separation screening
Achromobacter dolens M6, for remaining flubenzuron, diflubenzuron, d ichlorbenzuron benzene in degrade soil or water body environment
Benzoyl urea insecticide；The degradation bacterial agent of bacterial strain preparation can degrade a variety of benzoyl area kinds such as flubenzuron, diflubenzuron, d ichlorbenzuron
Insecticide, this microbial inoculum can make in a short time remaining flubenzuron in soil or water body environment, diflubenzuron, d ichlorbenzuron degradation rate
Up to 90% or more, flubenzuron in agricultural product, diflubenzuron, d ichlorbenzuron residual are eliminated, solves flubenzuron, diflubenzuron, d ichlorbenzuron to soil
The harm problem of earth water body environment, crops and human health.
Degradation bacterial agent of the invention can be produced with the general Zymolysis Equipment of fermentation industry, had production cost low, made
With convenient, the good advantage of removal effect, it is suitble to administer in soil and water body environment and is polluted caused by process for preparation of benzoylurea compounds；This
Invention reduces insecticide to crop phytotoxicity, protects the health of the people, improve the attached of agricultural product for preserving the ecological environment
It is value added to have great importance.
Detailed description of the invention
Fig. 1 is bacterial strain M6 bacterium colony photo；
Fig. 2 is bacterial strain M6 electromicroscopic photograph；
Fig. 3 is that the HPLC of bacterial strain M6 degradation flubenzuron analyzes map (A:ck；B: processing)；
Fig. 4 is that the HPLC of bacterial strain M6 degradation diflubenzuron analyzes map (A:ck；B: processing)；
Fig. 5 is that the HPLC of bacterial strain M6 degradation d ichlorbenzuron analyzes map (A:ck；B: processing).
Specific embodiment
Below in conjunction with drawings and examples, the invention will be further described.
Embodiment 1
The separation and identification of bacterial strain M6:
Haikou city Hainan profit standing grain pesticide is derived from for being enriched with the enriched medium of process for preparation of benzoylurea compounds degradation bacteria strains
The activated sludge of Co., Ltd's agricultural chemicals waste water biochemical treatment tank takes activated sludge 5.0g to be added in basal salt media, is added
50mg·L-1D ichlorbenzuron, 30 DEG C, 180rpm cultivates 7d, is transferred in identical culture medium with 5% inoculum concentration, continuous to transfer
3 times, gradient dilution pregnant solution takes 10-4~10-7Each 0.1mL of the pregnant solution of dilution is coated on added with 100mgL-1D ichlorbenzuron
Solid medium tablets on, after 30 DEG C of culture 4d, the single colonie that picking is grown is inoculated in containing 100mgL-1Various benzoyl ureas
In the culture medium of insecticides (flubenzuron, diflubenzuron, d ichlorbenzuron), 30 DEG C, 180rpm cultivates 5d, verifies its degradation effect.Base
Plinth salt culture medium formula (1L) are as follows: 5.0g glucose, 1.0gNH4NO3、1.0gNaCl、1.5gK2HPO4、0.5gKH2PO4、
0.2gMgSO4·7H2O.pH 7.0. solid medium adds 15.0g agar.
The verification method of degradation effect: isometric methylene chloride is added in culture solution and carries out full dose extraction, acutely shakes
Rear stratification is swung, after then taking the volatilization completely of 1mL lower layer methylene chloride, 1mL methanol dissolution (chromatographically pure) is added, uses filter membrane
(0.22 μm of aperture) filtering.Using the content of process for preparation of benzoylurea compounds in ultraviolet and high performance liquid chromatography measurement extracting solution, liquid
Phase chromatographic condition: mobile phase is methanol: water (80:20, V/V), and Zorbax C218 ODS Spherex reversed-phase column (5 μm, 4.6mm
× 250mm, Agilent, USA), column temperature is room temperature, and PDA detector measures wavelength 254nm, 10 μ L of sample volume, and flow velocity is
1.0mL·min-1.External standard method presses peak area quantification.
From pregnant solution, it is separated to 1 plant of process for preparation of benzoylurea compounds degradation bacteria, is named as M6, the bacterium is right in 3d
200mg·L-1Flubenzuron, diflubenzuron, d ichlorbenzuron degradation rate reach 90% or more.Bacterial strain M6 is raw on solid medium
After long 3d, bacterium colony dimpling, translucent, non-pigment, as shown in Figure 1；By negative staining, the bacterium is in bar under transmission electron microscope
It is as shown in Figure 2 not to form gemma for shape, amphitrichous, no pod membrane.Bacterial strain M6 is G?, size is 0.4-0.7 × 1.4-2.8 μm, aerobic
Growth, nitrate reaction are the positive, using glucose, gluconic acid, adipic acid, citric acid, have alkaline phosphatase activities,
Nitrite, indoles generation, gelatin liquefaction reaction negative.The form and physiological and biochemical property and Bergey ' s of bacterial strain M6
The description of achromobacter is consistent on manual of systematic bacteriology.
Using the genomic DNA of bacterial strain M6 as template, PCR amplification is carried out with bacterial 16 S rRNA gene order universal primer,
The 16S rDNA gene order that length is 1392bp is obtained, as shown in SEQ ID No:1, (Genbank accession number is
KX348283).Blast is carried out in RDP database (https: //rdp.cme.msu.edu/), the results showed that bacterial strain M6 and nothing
The homology of Chromobacterium (Achromobacter sp.) bacterial strain is nearest, with type strain Achromobacter dolens
26840 homology of LMG is greater than 99%, establishes phylogenetic tree, combining form and physiological and biochemical property, bacterial strain M6 is tentatively reflected
It is set to achromobacter (Achromobacter dolens).Bacterial strain M6 is delivered positioned at Wuhan, China, the China of Wuhan University
Type Tissue Collection (abbreviation CCTCC) preservation, deposit number are CCTCC NO:M 2016257, and the deposit date is 2016
On May 11, in.
Embodiment 2
Biodegradation test of the bacterial strain M6 to process for preparation of benzoylurea compounds in culture medium:
Final concentration of 100mgL is added in basal salt media (with embodiment 1) respectively-1Flubenzuron, diflubenzuron,
D ichlorbenzuron accesses bacterial strain M6 bacterium solution by the inoculum concentration of 1% volume ratio, while respectively in basal salt media (with embodiment 1)
Final concentration of 100mgL is added-1Flubenzuron, diflubenzuron, d ichlorbenzuron, by 1% volume ratio inoculum concentration inoculation inactivation bacterial strain
M6 is set as control 1, control 2, cultivates in 3,30 DEG C of constant-temperature tables of control, and timing sampling is tested using degradation effect in embodiment 1
Card method passes through degradation situation of the high performance liquid chromatography detection bacterial strain to flubenzuron, diflubenzuron, d ichlorbenzuron, the HPLC of degradation results
Analysis map is as shown in Figure 3-Figure 5, and degradation rate is shown in Table 1.
1 bacterial strain M6 of table is in the medium to the degradation effect of flubenzuron, diflubenzuron and d ichlorbenzuron
Bacterial strain M6 respectively reached 45.1%, 43.7% and to the degradation rate of flubenzuron, diflubenzuron, d ichlorbenzuron in 1 day
49.8%, 92.6%, 91.4% and 96.1% is respectively reached to the degradation rate of flubenzuron, diflubenzuron, d ichlorbenzuron in 2 days, is owned
The degradation rate of control is 0.
Embodiment 3
The preparation method of degradation bacterial agent
1) the cultured test tube liquid of bacterial strain M6 is inoculated in fermentation medium by the inoculum concentration of 1% volume ratio, oscillation training
It supports to logarithmic phase, fermenting microbe is made；
2) above-mentioned cultured fermenting microbe is inoculated with by the inoculum concentration of 8% (v/v, on the basis of culture volume) into dress
Liquid measure is cultivated in the culture medium for 500 liters of seeding tanks of 70% (on the basis of fermenter volume, similarly hereinafter), culture to logarithmic growth
Seed liquor is made in phase；
It 3) is 70% by the inoculum concentration access liquid amount of 8% (v/v, on the basis of culture volume, similarly hereinafter) by seed liquor
Production tank culture medium in cultivate.
4) ventilatory capacity of filtrated air is 1:0.8 in the incubation of seeding tank and production tank, and mixing speed is
170rpm, cultivation temperature are 28 DEG C, and entire process flow incubation time is 48 hours, and thalline quantity reaches 10 after fermentation
Hundred million/mL or more.Culture solution goes out tank and is directly distributed into liquid agent with Packaging Bottle after fermentation.
Wherein, the fermentation medium, the culture medium of seeding tank, the culture medium prescription of production tank are identical, are glucose
0.1wt%, NaCl1.0wt%, peptone 0.5wt%, yeast extract 0.25wt%, pH7.2.
Embodiment 4
The fermentation of degradation bacterial agent:
1) the cultured test tube liquid of bacterial strain M6 is inoculated in fermentation medium by the inoculum concentration of 1.5% volume ratio, is vibrated
Fermenting microbe is made to logarithmic phase in culture；
2) by above-mentioned cultured fermenting microbe by 10% (v/v, on the basis of culture volume) inoculum concentration be inoculated with into
Liquid amount is cultivated in the culture medium for 500 liters of seeding tanks of 70% (on the basis of fermenter volume, similarly hereinafter), and culture is raw to logarithm
For a long time, seed liquor is made；
It 3) is 70% by the inoculum concentration access liquid amount of 10% (v/v, on the basis of culture volume, similarly hereinafter) by seed liquor
Production tank culture medium in cultivate.
4) ventilatory capacity of filtrated air is 1:0.8, mixing speed 230 in the incubation of seeding tank and production tank
Rev/min, cultivation temperature is 32 DEG C, and entire process flow incubation time is 36 hours, after fermentation thalline quantity reach 1,000,000,000/
ML or more.Culture solution goes out tank and is directly distributed into liquid agent with plastic barrel after fermentation.
Wherein, the fermentation medium, the culture medium of seeding tank, the culture medium prescription of production tank are identical, are glucose
0.1wt%, NaCl1.0wt%, peptone 0.5wt%, yeast extract 0.25wt%, pH7.5.
Embodiment 5
The preparation method of degradation bacterial agent
The preparation method of 5 degradation bacterial agent of embodiment is substantially the same manner as Example 4, the difference is that in seed in step 4)
The ventilatory capacity of filtrated air is 1:0.9, mixing speed 200rpm, cultivation temperature 30 in the incubation of tank and production tank
DEG C, entire process flow incubation time is 42 hours.
Embodiment 6
Bacterial strain M6 is in the soil to the Degrading experiment of process for preparation of benzoylurea compounds
Vegetable garden soil is taken to be used as trying soil sample.Soil sample is crossed into 2mm sieve, takes a certain amount of flubenzuron, diflubenzuron and d ichlorbenzuron
Missible oil is dissolved in acetone, it is uniformly admixed in 1000g soil, keeps flubenzuron in soil, diflubenzuron and d ichlorbenzuron final concentration equal
For 10mgkg-1.Fresh bacterial strain M6 degradation bacterial agent, preparation method is referring to embodiment 5, after 5000rpm is centrifuged 5min, collects bacterium
Body, again with sterile aqueous suspension after washing 3 times with aseptic deionized water, adjusting cell concentration is about 1.0 × 109cfu·mL-1.With
10% inoculum concentration is linked into above-mentioned soil, is cultivated in 30 DEG C of constant temperature dark culturing casees, if not connecing the soil conduct pair of bacterium
According to during which the water-holding capacity of soil is maintained at 60%, after cultivating 10 days, high performance liquid chromatography is utilized to measure residual quantity.It is residual in soil
Verification method of the detection method of the flubenzuron, diflubenzuron and the d ichlorbenzuron that stay referring to the degradation effect of embodiment 1.Measurement result is
Bacterial strain M6 in 10 days to flubenzuron in soil, diflubenzuron, d ichlorbenzuron degradation rate respectively reach 91.5%, 90.2%,
92.1%.
Claims (10)
1. one plant of process for preparation of benzoylurea compounds degradation bacteria strains M6 is identified as achromobacter (Achromobacter dolens),
It is preserved in China typical culture collection center, the preservation time is on May 11st, 2016, and deposit number is CCTCC NO:
M2016257。
2. application of the bacterial strain M6 described in claim 1 in degradation process for preparation of benzoylurea compounds.
3. application according to claim 2, which is characterized in that bacterial strain M6 benzoyl urea in degradation soil or water body environment
Application in insecticides.
4. application according to claim 2 or 3, which is characterized in that the process for preparation of benzoylurea compounds is flubenzuron, removes
One of worm urea, d ichlorbenzuron are a variety of.
5. application of the bacterial strain M6 described in claim 1 in production process for preparation of benzoylurea compounds degradation bacterial agent.
6. a kind of degradation bacterial agent containing bacterial strain M6 produced using bacterial strain M6 described in claim 1.
7. degradation bacterial agent according to claim 6, which is characterized in that by made by following steps:
1) the cultured test tube liquid of degradation bacteria strains M6 is inoculated in fermentation medium by the inoculum concentration of 1-1.5% volume ratio, is shaken
Culture is swung to logarithmic phase, and fermenting microbe is made；
2) above-mentioned cultured fermenting microbe is inoculated in the culture medium of seeding tank by the inoculum concentration of 8-10% volume ratio, is cultivated
To logarithmic growth phase, seed liquor is made；
3) it is cultivated in the culture medium by seed liquor by the inoculum concentration access production tank of 8-10% volume ratio；
4) ventilatory capacity of filtrated air is 1:(0.8-1.0 in the incubation of seeding tank and production tank), mixing speed is
170-230rpm, cultivation temperature are 28-32 DEG C, and whole process incubation time is 36-48 hours, and it is straight to go out tank for culture solution after fermentation
It connects and is distributed into liquid agent with plastic barrel or Packaging Bottle.
8. application of the degradation bacterial agent as claimed in claim 6 in degradation process for preparation of benzoylurea compounds.
9. application according to claim 8, which is characterized in that degradation bacterial agent benzoyl in degradation soil or water body environment
Application in urea insecticide.
10. application according to claim 8 or claim 9, which is characterized in that the process for preparation of benzoylurea compounds be flubenzuron,
One of diflubenzuron, d ichlorbenzuron are a variety of.
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| CN113234438B (en) * | 2021-05-17 | 2023-09-01 | 西华大学 | Red light carbonized polymer dot based on citric acid and benzoyl urea, and preparation method and application thereof |
| CN114990019B (en) * | 2022-06-16 | 2023-07-18 | 上海市农业科学院 | Organic pollution degradation strain A7, microbial inoculum produced by same and application thereof |
| CN116478879B (en) * | 2023-04-11 | 2024-02-06 | 中国水产科学研究院 | Bacterial strain for degrading diflubenzuron and application thereof |
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