2,4-dinitrotoluene sulfonate high-efficient degradation strain Sphingomonas sp.X4 and application thereof
A kind of technology of dinitrotoluene sulfonate and strain, applied in the field of environmental biology
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Embodiment 1
[0024] Example 1: Isolation and purification of bacterial strain Sphingomonas sp.X4
[0025] Soil samples were collected from Baiyin City, Gansu Province (104°13′43.907″ east longitude, 36°30′44.676″ north latitude), packed in ziplock bags, stored at 4°C and shipped back to the laboratory.
[0026] Enrichment culture: Take 10g of contaminated soil and put it into a conical flask filled with 100ml LB liquid medium, shake it on a constant temperature bed at 30°C and 120rpm, and enrich it for 24 hours.
[0027] Acclimatization: Take 1ml of the above-mentioned culture solution and apply it to the solid inorganic salt medium containing 2,4-DNT-3-SA and 2,4-DNT-5-SA with a coating rod, and put it into a constant temperature incubator for 7 days;
[0028] Separation and purification: After the colony grows, observe the colony shape, use a sterile inoculation loop to select the colony with a large difference in shape from the above-mentioned solid inorganic salt medium, and inoculate ...
Embodiment 2
[0031] Example 2: Degradation Ability Determination of 2,4-Dinitrotoluenesulfonate Degrading Strain Sphingomonas sp.X4
[0032] Take uncontaminated soil, air-dry and grind, pass through a 1mm sieve, and set aside. A certain amount of 2,4-DNT-3-SA and 2,4-DNT-5-SA was weighed and dissolved in acetone. In the fume hood, the acetone solution of the two sulfonates was evenly sprayed into the above soil and stirred evenly. The soil concentration was about 2,4-DNT-3-SA and 2,4-DNT-5-SA concentrations of 500 mg / kg each; it was allowed to air dry naturally in a fume hood for 2 days.
[0033] A certain concentration of bacterial suspension was inoculated into the above soil according to the inoculum amount of 2%. And add a certain amount of inorganic salt liquid medium to maintain the water-to-soil ratio at 0.4. And as for culturing in a constant temperature incubator at 30°C, samples were taken every 24 hours, and the concentrations of 2,4-DNT-3-SA and 2,4-DNT-5-SA were determined ...
Embodiment 3
[0034] Example 3: The degradation effect of Sphingomonas sp.X4 on sulfonate at different pH values
[0035] Take uncontaminated soil, air-dry and grind, pass through a 1mm sieve, and set aside. A certain amount of 2,4-DNT-3-SA and 2,4-DNT-5-SA was weighed and dissolved in acetone. In the fume hood, the acetone solution of the two sulfonates was evenly sprayed into the above soil and stirred evenly. The soil concentration was about 2,4-DNT-3-SA and 2,4-DNT-5-SA concentrations of 500 mg / kg each; it was allowed to air dry naturally in a fume hood for 2 days. Weigh 20 g of the above soil into a conical flask, and use dilute H 2 SO 4 and NaOH to adjust the pH of the soil to be 3, 5, 7, 9, 11 respectively, add the bacterium liquid activated by the Sphingomonas sp.X4 of OD600=1 according to the inoculum size of 5%, and adjust the water-soil ratio with the inorganic salt liquid medium to be 2: 5. Seal with a breathable sealing film. All the Erlenmeyer flasks were placed in a cons...
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