Rhodococcus erythropolis for biological desulfurization and its use
A technology of Rhodococcus erythropolis and biological desulfurization, applied in the direction of bacteria, etc., can solve the problems of long growth cycle of strains, shortening the life of biocatalysts, and insufficient catalytic activity of desulfurization strains, and achieve good oil resistance, short growth cycle, and desulfurization highly active effect
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Embodiment 1
[0046] Embodiment 1: the screening of FSD-2 bacterial strain
[0047] Oil-contaminated soil samples were taken from the vicinity of the Gudao Oilfield in Shandong, and a three-step process was used to screen, isolate, and purify microorganisms that selectively desulfurize oil. In the first step, add 0.5 g of oil-contaminated soil sample to 50 ml of sulfur-free basic medium FBM containing a final concentration of 1 mmol / l DBT (FBM medium contains 10.0 g / l glucose, 2.0 g / l NH 4 Cl, 6.3g / l KH 2 PO 4 , 8.0g / l K 2 HPO 4 , 0.2g / lMgCl 2 ·6H 2 O, etc., the FBM medium containing 1mmol / l DBT is FBMD medium), 30 DEG C shaking culture 5 days, transfer to FBMD medium with 10% inoculum size, cultivate 2 days under the same condition, transfer 2 times repeatedly -3 times. In the second step, after such 2-3 enrichment cultures, take 1ml of the culture, centrifuge at 3000rpm for 5min, and take 0.5ml of the supernatant for Gibb's analysis (after DBT is degraded by the "4S" pathway, the g...
Embodiment 2
[0049] Example 2: FSD-2 strain metabolite analysis
[0050] Pick the FSD-2 strain cultured on the FMAD agar slant and inoculate into 50ml FMAD liquid medium. Components of FMAD liquid medium: 5g / l glucose, 2g / l NH 4 Cl, 14.04g / l Na 2 HPO 4 , 2.24g / l KH 2 PO 4 , 0.2g / l MgCl 2 ·6H 2 O, the addition amount of trace element solution mother solution and vitamin solution mother solution is 1.0ml / l respectively, the final concentration of DBT is 1.0mmol / l, and the pH value is 7.0~7.2. Trace element mother liquor contains 0.5g / l FeCl 2 4H 2 O, 0.5g / l ZnCl 2 , 0.5g / l MnCl 2 4H 2 O, 0.1g / l NaMoO 4 2H 2 O, 0.05g / l CuCl 2 , 0.05g / l NaWO 4 2H 2 O and 120mmol / l HCl. Vitamin mother solution contains 400mg / l calcium pantothenate, 200mg / l inositol, 400mg / l niacin, 400mg / l pyridoxine hydrochloride, 200mg / l p-aminotoluene and 0.5mg / l VB12. Culture at 30℃, 150 rpm After 24-48 hours, take 10ml of the culture, add 2ml of dichloromethane to extract the metabolites in the culture, a...
Embodiment 3
[0051] Embodiment 3: FSD-2 bacterial strain growth cell degrades DBT
[0052] Pick the FSD-2 strain cultured on the FMAD agar slant and inoculate into 50ml FMAD liquid medium. Components of FMAD liquid medium: 5g / l glucose, 2g / l NH 4 Cl, 14.04g / l Na 2 HPO 4 , 2.24g / l KH 2 PO 4 , 00.2g / l MgCl 2 ·6H 2 O, the addition amount of trace element solution mother solution and vitamin solution mother solution is 1.0ml / l respectively, the final concentration of DBT is 0.5mmol / l, and the pH value is 7.0~7.2. Trace element mother liquor contains 0.5g / l FeCl 2 4H 2 O, 0.5g / l ZnCl 2 , 0.5g / l MnCl 2 4H 2 O, 0.1g / l NaMoO 4 2H 2 O, 0.05g / l CuCl 2 , 0.05g / l NaWO 4 2H 2 O and 120mmol / l HCl. Vitamin mother solution contains 400mg / l calcium pantothenate, 200mg / l inositol, 400mg / l niacin, 400mg / l pyridoxine hydrochloride, 200mg / l p-aminotoluene and 0.5mg / l VB12. Culture at 30℃, 150rpm , regularly sampled for Gibb's analysis. Gibb's analysis method is as follows: centrifuge the cul...
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