DU-E4LP seudomonas putida and use in degradating parathionmethyl

A technology of methyl parathion and degrading bacteria, applied in bacteria and other directions, can solve problems such as health threats, and achieve the effect of large application potential and wide substrate utilization range

Inactive Publication Date: 2007-08-29
INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When this method degrades p-methyl parathion, the intermediate product produced is p-nitrophenol, which is a moderately toxic substance that will pose a potential threat to human health.

Method used

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  • DU-E4LP seudomonas putida and use in degradating parathionmethyl
  • DU-E4LP seudomonas putida and use in degradating parathionmethyl
  • DU-E4LP seudomonas putida and use in degradating parathionmethyl

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1, separation and screening methyl parathion degrading bacteria of the present invention

[0049] Sampling from the long-term pesticide-contaminated soil collected around Tianjin Pesticide Factory, the collected pesticide-contaminated soil sample is prepared into a 20wt% suspension with inorganic salt culture solution, and then received with 5wt% inoculum containing 50mg / L formazan Shake culture in the inorganic salt culture solution based on parathion; subculture for 6 generations, gradually increase the concentration of methyl parathion in the inorganic salt culture solution to 600ppm; Distribute on inorganic salt solid medium containing 500ppm methyl parathion, culture in a 30°C incubator; select colonies with fast growth (transparent circle on the plate), regular colonies, and stable passage, and purify 2 to 3 times to obtain pure Bacterial strain; The 5 strains that isolate are used as re-screening bacterial strains to expand and cultivate to the same OD...

Embodiment 2

[0060] Embodiment 2, identify the methyl parathion degrading bacteria of embodiment 1

[0061]The bacterial strains obtained in Example 1 were identified by using the computer classification and identification system for microorganisms (Biolog. Inc., USA). Cultivate the purified strain on Biolog medium for 16-24 hours; prepare the bacterial suspension with GN / GP-IF by dry tube method, adjust the turbidity to 52% T; inoculate the bacterial suspension with 150 μl per well on GN-Microplate for identification Incubate the identification plate at 30°C for 16-24 hours; scan the identification plate with Reader, measure the absorbance value of each well, compare the measured results with the database, and automatically give the identification results by the software. The strain was identified as Bacillus subtilis by Biolog technology, and named as methyl parathion degrading bacteria DM-1. The identification results are shown in Table 1.

[0062] Table 1. Identification results of m...

Embodiment 3

[0064] Embodiment 3, the influence of glucose concentration on the growth and degradation rate of methyl parathion degrading bacteria DM-1 of the present invention

[0065] The isolated methyl parathion-degrading bacteria DM-1 bacterial strain in LB medium (same as before) is enlarged and cultivated OD value about 1.5, washes with sterile water after centrifugation, prepares bacterium suspension, with 10% inoculum size is inserted in the inorganic salt culture fluid (the same as before) that contains methyl parathion 200ppm, adds the inorganic salt liquid culture fluid of the glucose of different concentrations respectively (the concentration of glucose is respectively 0g / L, 1g / L L, 2g / L, 5g / L, 10g / L, 20g / L, pH value of 7.0), and with no inoculation as a control, two parallels are set for each sample. 30°C, 180r / min shaker shaking culture for 32h. Get 10ml of culture solution, extract 3 times with equal volume of ethyl acetate, combine three extracts, dry over anhydrous sodiu...

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Abstract

The invention supplies a methyl parathion degradation fungus DM-1 that the preservation NO is CGMCC No.1620. The classification designation is Bacillus subtilis. It is separated and selected from soil polluted by agricultural chemical. The fungus has good degradation effect to 50-500ppm methyl parathion at 30 degree centigrade.

Description

technical field [0001] The invention belongs to the field of microbial degradation, and in particular relates to a methyl parathion degrading bacterium and its application in degrading methyl parathion. Background technique [0002] Methyl Parathion (MP for short) is a highly toxic organophosphorus insecticide with the chemical formula C 8 h 10 NO 5 PS, the chemical name is O, O-dimethyl O-(p-nitrophenyl) phosphorothioate. Due to the characteristics of high efficiency and economy, this insecticide is widely used in agricultural production and pest control. But at the same time, because its residual poison will cause serious pollution to the environment and food, and produce a large amount of pesticide waste, seriously affect people's health and the quality of export agricultural products, bringing a series of negative effects. How to effectively deal with the residual methyl parathion to avoid damage to the environment and human beings has become an urgent problem to be ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20
Inventor 乔传令杨超董明黄瑶
Owner INST OF ZOOLOGY CHINESE ACAD OF SCI
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