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High efficiency chlopyrifos degradating strain capable of generating magic spots

A technology of chlorpyrifos and strains, applied in the field of microorganisms, can solve the problems of low degradation ability, few strains, easy degradation of bacterial degradation ability, etc., and achieve the effects of in-depth research on metabolism, stable degradation performance, and strong degradation ability.

Inactive Publication Date: 2012-04-04
中国农业科学院研究生院
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Problems solved by technology

[0004] However, due to the few strains isolated at home and abroad, the degradation ability is low, the degradation ability of strains is easy to degrade after multiple passages, and the research on the metabolic regulation mechanism of degrading strains is few, so far, the bioremediation of organophosphorus pesticide pollution has still a certain distance

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  • High efficiency chlopyrifos degradating strain capable of generating magic spots

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Experimental program
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Effect test

Embodiment 1

[0032] Embodiment 1 bacterial strain enrichment culture method

[0033] 1) Enrichment culture: Take 1.0 g of activated sludge in the pesticide sewage treatment tank and add it to the enrichment culture containing chlorpyrifos 100mg / L. Based on 30°C, 200rpm enrichment culture for 7 days;

[0034] 2) Increase the selection pressure: transfer the above-mentioned enrichment solution to a fresh enrichment medium with an inoculum size of 10%, and double the pesticide concentration, and then continue to cultivate for 7 days, and transfer 6 times in this way until the enrichment culture The pesticide concentration in the base was increased to 800mg / L.

[0035] Wherein, the composition of the enrichment medium TYC in the step 1) is as follows: tryptone 5g / L, yeast extract 5g / L, KH 2 PO 4 1g / L and agar 15g / L, pH 7.0

Embodiment 2

[0036] Embodiment 2 Bacterial strain domestication isolation method

[0037] 1) Acclimatization culture: After the enrichment culture, 10% of the bacteria were collected by centrifugation, and then transferred to the inorganic salt medium respectively, and 600 mg / L of chlorpyrifos was added, and domesticated for two weeks under the same culture conditions; and different solvents ( Methanol, acetone, ethanol, pyridine) chlorpyrifos mother solution, to exclude the dependence of the strain on the solvent.

[0038] 2) Separation and purification: spread the acclimatized bacterial solution on an inorganic salt medium plate containing 300 mg / L chlorpyrifos by plate dilution method, and select a strain that grows well on the pesticide-containing medium, is stable in passage and has good degradation ability Carry out single bacterium colony streak separation, finally obtain the highly efficient chlorpyrifos degrading bacterial strain Klebsiella sp.CPK of purification (its preservation...

Embodiment 3

[0041] Example 3 Bacterial Strain Degradation Chlorpyrifos Detection Method

[0042] 1) TYC solid plate culture: Pick a single colony of the Klebsiella sp.CPK strain obtained in Example 2, activate it on TYC medium, and culture it at a constant temperature of 30°C until a single colony that can be picked grows.

[0043] Wherein, the enrichment medium TYC is the same as in Example 1

[0044] 2) Seed liquid culture: pick the above-mentioned activated single colony and inoculate the TYC medium (same as step 1), and culture at a constant temperature of 30°C until the logarithmic growth phase.

[0045] 3) Sampling for degradation: Inoculate the above seed solution with 20% inoculation amount into the inorganic salt MSM culture solution containing 50 mg / L chlorpyrifos as the sole carbon source, cultivate at 30°C, and take samples at different time points, three In parallel, use n-hexane equal volume vortex to extract chlorpyrifos residue, vortex for 2 minutes, and stand at 4°C for ...

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Abstract

The invention relates to a high efficiency chlopyrifos degradating strain Klebsiella sp. CPK capable of generating magic spots, and the accession number of the strain is CGMCC No.4167. The invention relates a preparation method and application of the strain. The strain provided in the invention can be widely used for removal of residual pollution of chlopyrifos and for bioremediation and has the advantages of powerful degradation capability, stable degradability, in-depth metabolic studies, etc.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to a high-efficiency chlorpyrifos-degrading bacterial strain capable of producing magic spots. Background technique [0002] At present, many strains of chlorpyrifos-degrading bacteria have been isolated by enrichment separation at home and abroad. For example, Anwar et al. have isolated a strain of Bacillus pumilus strain C2A1 (Bacillus pumilus strain C2A1) from soil by enrichment separation. Chlorpyrifos degraded 73% within 10 days (J Hazard Mater, 2009, 168: 400-405); Xu Gangming et al. also used the enrichment separation method to isolate several strains of chlorpyrifos-degrading bacteria from the activated sludge of pesticide factories and cultured them purely or by mixing The strains were cultured to detect the degradation of chlorpyrifos (Biotechnol Lett, 2007, 29: 1469-1473; International Biodeterioration & Biodegradation, 2008, 62: 51-56). [0003] Recently, magic spot accumu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A62D3/02C12R1/22A62D101/04A62D101/28
Inventor 闫艳春王圣惠李康史延华翟逸曲杰
Owner 中国农业科学院研究生院
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