Rhodococcus sp.P52 capable of degrading dibenzofuran and use thereof
A rhodococcus and diphenyl technology, applied in the biological field, can solve the problems of lack of degradation ability to degrade strains, difficulty in removal, poor adaptability, etc., and achieve the effects of strong processing ability and adaptability, fast degradation rate, and improved adaptability
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Embodiment 1
[0043] Example 1. Screening of Rhodococcus sp.P52CCTCC No: M2011181 strain
[0044] Collect soil samples from oil-contaminated areas such as Shengli Oilfield, take 1 g and add them to inorganic salt liquid medium containing 200 mg / L dibenzofuran, and culture them in a reciprocating shaker at 30°C for 3-5 days. Take 5% (v / v) of the total volume of the culture solution and transfer it to fresh culture solution, repeat 3 times, inoculate the culture solution on the inorganic salt solid medium containing 200mg / L dibenzofuran and separate it by streaking, 30 Cultivate at ℃ for 2-4 days, pick colonies with clear edges, and repeatedly purify by streaking on the plate until pure bacteria are obtained, and verify their ability to degrade dibenzofuran and 2-chlorodibenzofuran. The result is as figure 1 , figure 2 shown.
[0045] The formula of inorganic salt liquid medium is: K 2 HPO 4 12g, KH 2 PO4 11g, Na 2 SO 4 2.0g, MgSO 4 ·7H 2 O0.2g, NH 4 NO 3 3g, trace metal salt ...
Embodiment 2
[0047] Embodiment 2, the preparation of Rhodococcus sp.P52 bacterium liquid
[0048] (1) Select Rhodococcus sp.P52, CCTCC No: M2011181;
[0049] (2) Slant culture: inoculate a ring of single colonies in a solid slant medium (containing 1.5 wt% agar) containing a concentration of 500 mg / L dibenzofuran inorganic salt, and culture at 30° C. for 48 hours;
[0050] (3) Seed culture: inoculate 2 loops of the strain cultivated in step (2) into 50 mL of inorganic salt liquid medium containing 500 mg / L dibenzofuran under sterile conditions, and culture at 30°C with shaking at 180 rpm / min 36 hours made seed liquid;
[0051] (4) Expansion culture: Inoculate the seed liquid into 500 mL dibenzofuran (concentration 500 mg / L) inorganic salt liquid medium with an inoculum size of 5% (v / v), and culture with shaking at 180 rpm / min at 30°C for 48 Hour;
[0052] (5) Collecting cells: centrifuging the culture solution of step (4) at 5000 rpm for 10 minutes, collecting the expanded cultured cell...
Embodiment 3
[0054] Embodiment 3, degradation experiment of dibenzofuran
[0055] Add dibenzofuran to the cell suspension prepared in Example 2, so that the final concentration of dibenzofuran reaches 500mg / L, shake at 180rpm / min at 30°C, take samples during the treatment, and detect dibenzofuran by gas chromatography The degradation situation, when the dibenzofuran in the mixed solution is completely undetectable, continue to cultivate for 1 hour and then terminate the treatment.
[0056] Described gas chromatography adopts Agilent 6890 type gas chromatograph, and chromatographic column is HP-5 capillary column (30m * 250 μ m * 0.25 μ m), and helium is used as carrier gas, and instrument analysis condition is: FID detector, injector, detection The temperature of the oven is 280°C, and the temperature of the column is 170°C. The samples taken during the treatment were treated by the following method and then determined by gas chromatography for the determination of dibenzofuran. Centrifu...
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