Oil-degrading surfactant-producing plant growth-promoting bacterium and application thereof
A technology for producing surfactants and surfactants, which is applied in the field of plant growth-promoting bacteria producing surfactants, and can solve the problem of less Acinetobacter
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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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