Pseudomonad strain and application thereof in degrading petroleum pollutants

A technology of Pseudomonas and petroleum, applied in the field of Pseudomonas and its application in the degradation of petroleum pollutants, can solve the problem of low bioremediation value and achieve the effect of high bioremediation value

Inactive Publication Date: 2015-12-23
CHINA UNIV OF PETROLEUM (EAST CHINA)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Dibenzothiophene is used as a model of sulfur-containing heterocyclic aromatic hydrocarbons. The metabolic pathways of microorganisms to dibenzothiophene are mainly divided into "4S" pathway (the produc

Method used

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  • Pseudomonad strain and application thereof in degrading petroleum pollutants
  • Pseudomonad strain and application thereof in degrading petroleum pollutants
  • Pseudomonad strain and application thereof in degrading petroleum pollutants

Examples

Experimental program
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Effect test

Embodiment 1

[0036] Example 1: Dibenzothiophene degradation

[0037] The Pseudomonassp.LKY-5 strain was inoculated into 100 mL of an inorganic salt medium containing 100 mg / L dibenzothiophene. 30℃, pH7.5, inoculation amount 10wt%, 160rpm shaker culture for 7 days, after acidification with 6M HCl to pH≤2, add an equal volume of ethyl acetate for extraction, centrifuge at 4℃, 5000rpm for 10min, take the supernatant The liquid was measured by Bruker450 gas chromatograph (FID, HP5 capillary column), and the degradation rate of dibenzothiophene reached 100%.

Embodiment 2

[0039] The Pseudomonassp.LKY-5 strain was respectively inoculated into an inorganic salt medium containing 100 mg / L of phenanthrene, pyrene, carbazole, and n-hexadecane at 30°C, pH 7.5, inoculation volume 10 wt%, and 160 rpm shaker culture 7 In days, n-hexadecane was measured by Bruker450 gas chromatograph. Phenanthrene, pyrene and carbazole were determined by high performance liquid chromatography Waters2695, using UV detector, Waterssunfire C18 reversed phase chromatography column (column length 150mm, inner diameter 4.6mm, packing 5μm), the column was kept at 25℃, and the absorption peak of the substance was detected at 254nm Mobile phase A is water containing 0.5% trifluoroacetic acid, and mobile phase B is acetonitrile containing 0.5% trifluoroacetic acid. Using gradient elution, see Table 1.

[0040] Table 1 Gradient elution program

[0041]

[0042] The result is figure 2 As shown, Pseudomonassp.LKY-5 has a degradation effect on n-hexadecane, phenanthrene, pyrene and car...

Embodiment 3

[0044] Add 2mL of commercially available No. 0 diesel (density 0.841g / mL) and 2mL of Qingdao Refinery Catalytic Cracking Diesel (density 0.9468g / mL) to the inorganic salt medium (the final volume of the medium is 30mL), Pseudomonassp.LKY The inoculation amount of strain -5 was 10wt%, and the uninoculated strain was used as a blank control. 30℃, 160rpm, 7 days after incubation time, the blank control and the degraded oil-containing medium were extracted with petroleum ether (30~60℃) by shaking, and the extracts were combined, centrifuged at 5000rpm, 4℃ for 10min, and subjected to anhydrous sulfuric acid After the sodium is dehydrated, it is rotary evaporated under reduced pressure at 30°C until the petroleum ether is completely volatilized, then transferred to a chromatographic bottle, and analyzed by a US Varian 3800 gas chromatograph. The detector is a pulsed flame photometric detector (PFPD), HP-5NS quartz capillary column (30m×0.32mm×0.25μm), injection volume 1μL, split rati...

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Abstract

The invention relates to the field of application of microbes in environmental pollutant degradation, particularly a pseudomonad strain and application thereof in degrading petroleum pollutants. The pseudomonad strain (Pseudomonas sp. LKY-5) is screened from petroleum polluted soil in Shengli Oil Field in China, and the collection number is CCTCC NO:M2015448. The screened Pseudomonas sp. LKY-5 can effectively degrade sulfur-containing heterocyclic aromatics, such as dibenzothiophene, benzothiophene, alkyl dibenzothiophene, 4,6-dimethyl dibenzothiophene, 2,4,6-trimethyl dibenzothiophene and the like. Meanwhile, the Pseudomonas sp. LKY-5 can degrade many other pollutants in crude oil, such as n-hexadecane, phenanthrene, pyrene, carbazole and the like.

Description

Technical field [0001] The invention relates to the application field of microorganisms in the degradation of environmental pollutants, in particular to a strain of Pseudomonas and its application in the degradation of petroleum pollutants. Background technique [0002] Petroleum pollutants are the most important pollutants produced during oilfield development and petroleum processing. The hazards of petroleum pollutants are mainly manifested in the hazards and impacts on humans, animals, soil and natural water bodies. Petroleum pollutants have been included in the list of hazardous wastes in my country, and among the 48 hazardous wastes listed, petroleum ranked 8th. [0003] Blowouts, oil tank leaks, oil tanker leaks, oilfield equipment maintenance, petrochemical production, transportation accidents, etc., will overflow and discharge petroleum hydrocarbons. Among them, sulfur-containing heterocyclic aromatic hydrocarbons are an important component of high-sulfur crude oil, and m...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34B09C1/10C12R1/38C02F101/30
Inventor 赵朝成李琳刘芳刘其友张云波刘春爽陈水泉
Owner CHINA UNIV OF PETROLEUM (EAST CHINA)
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