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Bacillus subtilis and its application

A technology of Bacillus subtilis and bacterial strains, applied in bacteria, petroleum industry, refined hydrocarbon oil, etc., can solve problems such as limited desulfurization capacity, and achieve the effect of strong desulfurization function

Active Publication Date: 2005-04-27
CHINA PETROLEUM & CHEM CORP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Although people have isolated various desulfurization microorganisms, their desulfurization ability is limited, and they have to undergo mutagenesis screening or molecular level improvement, or provide new types of desulfurization microorganisms

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Screening method for Bacillus subtilis B. Subtilis.

[0025] (1). Take 10ml of basic salt medium, add 2mM DBT, and insert 5g of soil sample. Shake for 1 minute and settle for 30 minutes. Use a pipette to take out the supernatant and put it into the broth medium, and cultivate it at 28-30°C for 24-48 hours.

[0026] (2). The above-mentioned cultured strains were inoculated into the selection medium with DBT as the only sulfur source, and cultured at 30° C. for 48 hours. After further enrichment and cultivation for 2-3 days, the culture medium was taken and centrifuged, and the supernatant was analyzed by Gibb's. The blue color indicated that the culture contained a strain that could specifically convert DBT into 2-HBP, or it could be analyzed by gas chromatography / mass spectrometry. Analyze and detect 2-hydroxybiphenyl to judge the existence of specific desulfurization bacteria.

[0027] (3). The desulfurization microorganisms that were positive in the prev...

Embodiment 2

[0029] Example 2: Ultraviolet mutagenesis screening was carried out on the screened B. Subtilis strains.

[0030] With the B.Subtilis bacterial strain isolated in Example 1 as the starting strain, through ultraviolet (U.V) mutagenesis, the mutant strain B.Subtilis Fds-1 with strong desulfurization ability is screened, the method is as follows:

[0031] (1). Determining the lethality rate of ultraviolet radiation

[0032] The isolated Bacillus subtilis with desulfurization ability was inoculated into the enrichment medium of basal salt plus DBT, and cultured with shaking at 28-30°C for 2 days. The above culture medium was centrifuged to collect the cells. After washing with normal saline, the bacterial suspension was prepared. Take 10ml of bacterial suspension in a petri dish with a magnetic rotor, put it under a 20w ultraviolet lamp, and irradiate it at a vertical distance of 30cm for 6 minutes. Samples are taken every 30 seconds. Bacterial liquids with different irradiati...

Embodiment 3

[0038] Example 3: Growth of B. Subtilis Fds-1 strain Desulfurization of cells.

[0039] (1) Insert the B.Subtilis Fds-1 bacterial strain into 50ml basal salt medium containing 0.5~1.0mM DBT according to 10% inoculum size (the components are: every liter of distilled water contains 10.0g glucose, 4.0g NH 4 Cl, 6.3g KH 2 PO 4 , 8.0 g K 2 HPO 4 , 0.2g MgCl 2 ·6H 2 O, 1.0ml metal salt solution and 1.0ml vitamin mixture. Among them, the metal salt solution is 2.0g CaCl 2 , 1.0g NaCl, 0.5g FeCl 2 4H 2 O, 0.5g ZnCl 2 , 0.5g MnCl 2 4H 2 O, 0.1 g Na 2 MoO 4 2H 2 O, 0.05g CuCl 2 , 0.05g Na 2 WO 4 2H 2 O and 10 ml of 10M HCl in 1 liter of distilled water; the vitamin mix is ​​400 mg calcium pantothenate, 200 mg inositol, 400 mg niacin, 400 mg pyridoxine hydrochloride, 200 mg p-aminobenzoic acid, and 0.5 mg vitamin B 12 solution in 1 liter of distilled water). Cultivate on a shaker at 30° C. and 200 rpm for 48 hours, and measure the concentration of bacteria in the cul...

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Abstract

The present invention relates to Bacillus subtilis Fds-1 with the preservation number of CGMCC No. 0644. The strain of the present invention is obtained through separation from the soil near high sulfur oil well, ultraviolet mutagensis and screening. The growing cell and the rest cell of the strain can make the C-S bond oxidized to break and thus eliminate organic sulfur from organic compound, so that the strain is especially suitable for eliminate organic sulfur from petroleum fractions in high desulfurizing rate.

Description

technical field [0001] The invention relates to a bacillus subtilis and its application in deep desulfurization of petroleum distillate oil. Background technique [0002] When the sulfides contained in fossil fuels are burned, a large amount of toxic gases such as sulfur dioxide are released, and the resulting large-scale acid rain seriously pollutes the air and water sources, destroys the ecological balance, and endangers human existence. The composition of organic sulfur compounds in fossil fuels is complex, and most of them are heterocyclic compounds. The C-S chemical covalent bond is very strong. It is difficult to effectively remove organic sulfur in heterocyclic rings by conventional physical and chemical methods, and the cost is also high. [0003] In recent years, the rapidly developing biodesulfurization (BDS) technology in the world will become an effective way to reduce the sulfur content of petroleum products. BDS can be widely used in the treatment of gasoline,...

Claims

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Application Information

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IPC IPC(8): C10G32/00C12N1/20C12S1/02
Inventor 佟明友方向晨张全李淑兰
Owner CHINA PETROLEUM & CHEM CORP
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