A surfactant-producing plant growth-promoting bacterium for degrading oil and its application
A technology for producing surfactants and surfactants, applied in the direction of bacteria, microorganisms, water pollutants, etc., to achieve the effects of promoting plant growth, broad application prospects and commercial value, and the ability to produce surfactants
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Embodiment 1
[0028] Screening and cultivation of embodiment 1 bacterial strain S4
[0029] (1) Sample source: oily soil in a coking plant in Hubei Province.
[0030] (2) Preliminary screening of bacterial strains: Dilute the soil extract to 10 with sterile water according to the concentration gradient. -3 ~10 -8 , and then take 100-200 μL and evenly spread it on the LB solid plate, select the bacterial cells of different colors and shapes and inoculate them on a new LB solid plate by the plate streak method for separation and purification.
[0031] (3) Screen strains with plant growth-promoting characteristics: Inject the purified single colonies into LB medium containing tryptophan (tryptone 10 g, yeast extract 5 g, sodium chloride 5 g, L-tryptophan 0.1 g, add water to 1L, pH 7.0-8.0) and tricalcium phosphate medium (glucose 10g, Ca 3 (PO 4 ) 2 5g, MgCl 2 5g, MgSO 4 ·7H 2 O 0.2g, KCl 0.2g, (NH 4 ) 2 SO 4 0.2g, add water to 1L, pH 7.0~8.0), culture at 28°C, 150rpm and set a s...
Embodiment 2
[0035] Identification and category determination of embodiment 2 bacterial strain S4
[0036] The physiological, biochemical and genetic characteristics of strain S4 were observed and identified according to the following methods.
[0037] (1) Observed by naked eye and optical microscope, the single cell of strain S4 is spherical, without flagella, and without spores, often in a double distribution. The colonies are round, with neat edges, smooth and moist surface, and light yellow, such as figure 1 shown.
[0038] (2) Physiological and biochemical properties: Gram-negative bacteria, oxidase negative, motility negative, nitrate negative, hemolysis positive, gelatin hydrolysis positive.
[0039] (3) Genetic characteristics: In order to determine the type of strain S4, 16S rDNA gene sequencing was used for identification. First, bacterial DNA was extracted using the D3350-01 bacterial DNA kit (Omega Biotek, USA). Two sets of 16S rDNA bacterial PCR universal primers: 27F [5'-A...
Embodiment 3
[0042] Example 3 Bacterial Strain S4 Degradation Ability Measurement to Different Oily Sewages
[0043] According to the activation of bacterial strain S4 in Example 1 and the preparation of seed liquid, fermentation was carried out as follows.
[0044] (1) Sewage containing diesel oil: add 5000mg / L diesel oil to 20mL sterilized fermentation medium (KH 2 PO 4 0.5g, K 2 HPO 4 1.5g, (NH 4 ) 2 SO 4 1.0g, MgSO 4 ·7H 2 O 0.1g, NaCl 1.0g, add water to 1L, pH 7.0), and then insert Acinetobacter S4 seed liquid with an inoculum size of 5% (v / v); after culturing at 28°C and 150rpm for 6 days, add 20mL Ultrasonic extraction of petroleum ether for 15 minutes, dehydration with anhydrous sodium sulfate, and colorimetry at a wavelength of 225nm. The results are shown in Table 1, showing that strain S4 can degrade 5000 mg / L of high-concentration diesel-containing sewage to 1864 ± 200 mg / L after 6 days, and the degradation rate is about 62.7%, indicating that bacterial strain S4 ha...
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