Gordona terrae C-6 and its desulfurization effect
A strain and technology of desulfurization bacteria, applied in the field of auxiliary fuel oil deep desulfurization, can solve the problem that Rhodococcus cannot be removed
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Embodiment 1
[0043] Embodiment 1 Screening steps of the Gordona terrae C-6 bacterial strain provided by the present invention
[0044] Collect oil-soaked soil samples from around the high-sulfur oil wells in Gudao Oilfield, Shandong, China, and take 5g of samples suspended in 250mL sulfur-free medium WS in a 250mL triangular flask (WS medium composition: 1000mL deionized water contains: 2.56g of K 2 HPO 4 , 2.08 g KH 2 PO 4 , 1.00 g NH 4 Cl, 0.25g MgCl 2 , 0.001g CaCl 2 , 0.005g of triammonium citrate, 5.00g of sodium succinate, pH7.2), 10 to 20 glass beads are placed in the Erlenmeyer flask to break up the soil sample suspension, and the medium is pre-autoclaved at 121°C for 25 minute. Put it in an air-bath shaker and mix for 30 minutes. After standing still, take 1 mL of the upper layer liquid and add it to the enrichment medium (the composition is the above-mentioned basal salt medium plus 0.1-0.3% sulfur-free carbon source and 0.01%-0.05% BT). After 2-3 days of culture at 30°C ...
Embodiment 2
[0046] Example 2 PCR amplification, sequencing and comparison of the 16S rRNA sequence of the Gordona terrae C-6 strain of the present invention 1, a small amount of bacterial total DNA extraction
[0047] ①Pick a freshly activated single colony on the LB plate and put it in 5mL LB medium, shake and culture at 37°C for 12 hours;
[0048] ②Transfer 3mL of bacterial liquid to a 5mL centrifuge tube, centrifuge at 4°C, 12,000 rpm for 5 minutes, and discard the supernatant;
[0049] ③Wash the bacterial sediment once with 0.5mol / L NaCl, and dry the supernatant as much as possible;
[0050] ④Use liquid nitrogen to fully grind the bacterial sediment into powder;
[0051] ⑤Resuspend the ground bacteria in 1mL 50mmol / L Tris (pH 8.0) buffer, then add 0.2mL freshly prepared lysozyme solution (10mg / mL in 0.25mol / L Tris, pH8.0) and 0.8mL 0.25mol / L EDTA solution, mix well and treat at 37°C for 1 hour, then add 200μL 10% SDS solution, mix well and place at 55°C for 5 minutes;
[0052] ⑥ Ad...
Embodiment 3
[0092] Example 3 Preparation of cell liquid culture of Gordona terrae C-6 bacterial strain
[0093] Pick a single colony of Gordonia C-6 cultured on a nutrient agar plate and put it in a test tube containing 5 mL of sterilized basic inorganic salt medium WS-yeast powder, and cultivate it on a shaker at 180 rpm for 36-48 hours. Draw 1mL of the bacterial solution in the test tube and add it to a 500mL Erlenmeyer flask containing 100mL of basic inorganic salt medium WS, then add sterilized BT-ethanol stock solution (to make the BT concentration 0.5mmol / L) and the final concentration is 1% sterile dextrose stock solution. The Erlenmeyer flask was placed in a shaker at 30° C. and cultured at 180 rpm for 48 hours to prepare a liquid culture solution of Gordonia C-6 cells. Bacteria concentration was determined by the dry weight of cells and the optical density (OD) of cell suspension at 660nm 660 ) The linear relationship between ) was obtained by measuring the OD value of the cult...
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