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Microbacterium oxydans for degrading polycyclic aromatic hydrocarbon and application thereof

A technology for oxidizing microbacteria and polycyclic aromatic hydrocarbons, which can be used in the fields of microbial biotechnology and environmental biology, and can solve the problems of high content of polycyclic aromatic hydrocarbons and refractory crude oil components.

Active Publication Date: 2015-02-11
PETROCHINA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the past, microbial oil recovery strains were mainly screened to degrade saturated hydrocarbons such as long-chain alkanes, cycloalkanes, and waxes. However, in some heavy oil reservoirs, the content of macromolecular or ultra-macromolecular polycyclic aromatic hydrocarbons is high, and it is difficult for saturated hydrocarbon degrading bacteria to degrade saturated hydrocarbons. degrade these crude oil components

Method used

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  • Microbacterium oxydans for degrading polycyclic aromatic hydrocarbon and application thereof
  • Microbacterium oxydans for degrading polycyclic aromatic hydrocarbon and application thereof
  • Microbacterium oxydans for degrading polycyclic aromatic hydrocarbon and application thereof

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Experimental program
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Effect test

Embodiment 1

[0043] Embodiment 1: Screening and breeding of microbacterium oxydans (Microbacterium oxydans) Ji 2 bacterial strain provided by the present invention

[0044] Take a water sample from Zhongyuan Oilfield, oscillate evenly, take 10ml with a pipette, and inoculate aseptically into 100ml inorganic salt basic medium (culture medium composition g / L: heavy oil 5, K 2 HPO 4 1, KH 2 PO 4 1. NaNO 3 4. MgSO 4 0.5, (NH 4 ) 2 SO 4 2. Yeast powder 0.2, distilled water, 1000ml, pH7.2, sterilized at 121°C for 30min, cultured on a shaker at 35°C for 7 days. Dilute the enriched shake flask with sterile water to 10 -5 、10 -6 、10 -7 , spread on new Wang's + liquid wax and LB plates, culture at 35°C for 2 days, and select strains with different colony shapes and sizes. Observe the colonies that grow. Single colonies were picked and checked for purity using a microscope. Inoculate each colony grown at 35°C into nutrient broth medium and cultivate to OD 600 It is about 0.8, and it is in...

Embodiment 2

[0045] Embodiment 2: Morphological characteristics and physiological and biochemical characteristics of the microbacterium oxydans (Microbacterium oxydans) Ji 2 bacterial strain provided by the present invention

[0046] Carry out with reference to the experimental method of "Bergey's Manual of Systematic Bacteriology" (Vol.Ⅷ), detect the bacterium colony morphology of Ji 2 of the present invention, Gram staining, thalline size and shape, presence or absence of flagella and spores, growth temperature, growth pH range , NaCl tolerance. Catalase, V-P, mouse acute oral, casein hydrolysis, gelatin hydrolysis, starch hydrolysis, nitrate reduction to nitrite, cellulose hydrolysis, glucose gas production, tyrosine hydrolysis, phenylalanine dehydrogenase, Yolk lecithinase, NaCl and KCl requirement, allantoin and urea salt requirement, oxidase, lysozyme resistance, D-glucose, L-arabinose, D-xylose, D-mannitol acid production, citrate and propionate requirements etc. experiments.

[0...

Embodiment 3

[0048] Embodiment 3: The present invention provides the PCR amplification and sequence determination of the 16S rRNA gene of Microbacterium oxydans (Microbacterium oxydans) Ji 2 bacterial strain

[0049] The Ji 2 bacterial strain of the present invention was inoculated in LB medium, cultured on a shaker at 35°C (120rpm) for 24h, centrifuged to collect the bacteria, resuspended, added lysozyme and SDS to break the wall, extracted genomic DNA by the phenol-chloroform method, and used Forward primer 8F (5'-AGA GTT TGA TCC TGG CTC AG-3', SEQ ID No.2) and reverse primer 1541R (5'-AAGGAGGTGATCCA GCC-3', SEQ ID No.3), use this pair The primers were used to amplify the 16SrDNA gene by PCR, and the amplified primers were sent to Shanghai Meiji Company for sequencing. The PCR conditions are: 94°C, 10min; 94°C, 45s, 55°C, 45s, 72°C, 90s, 30 cycles; 72°C, 10min, 4°C storage. The length of the 16S rDNA gene sequence is 1563bp (SEQ ID No.1), the accession number in GenBank is KJ627769, and...

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Abstract

The invention provides a microbacterium oxydans for degrading a polycyclic aromatic hydrocarbon and the application thereof. Particularly, the invention provides a microbacterium oxydans Ji2 strain with a preservation number of CGMCC No.9072 and the application effect of the microbacterium oxydans in emulsifying and degrading crude oil and the polycyclic aromatic hydrocarbon and performing indoor physical simulation oil displacement. According to the microbacterium oxydans strain, the crude oil can be emulsified and degraded under aerobic and anaerobic conditions by taking the crude oil or the polycyclic aromatic hydrocarbon as the only carbon source and energy source, the degradation rate of the degraded polycyclic aromatic hydrocarbon, such as naphthalene is up to 100 percent, and analysis on the degraded crude oil shows that aromatic hydrocarbon and colloid, which are the components of the crude oil, are mainly degraded. The stain can be used for degrading the polycyclic aromatic hydrocarbon in an environment by the performance of the stain, and can also be applied to microbial enhanced oil recovery, and the oil recovery is improved.

Description

technical field [0001] The invention belongs to the field of microbial biotechnology and environmental biotechnology. Specifically, the invention relates to a strain of Microbacterium oxydans Ji 2 with a preservation number of CGMCC No.9072 and its use for emulsifying and degrading crude oil and polycyclic aromatic hydrocarbons and Indoor physical simulation of oil displacement. Background technique [0002] At present, most oilfields in my country have entered the stage of high water-cut development. The remaining recoverable reserves of crude oil are insufficient, and the difficulty is increasing. The main oilfields that have been developed will face the severe situation of total production decline. Therefore, enhancing oil recovery has always been a widely concerned issue in the world's oil recovery industry. In recent years, microbial oil recovery, as an economical and effective replacement technology, has received widespread attention, developed rapidly, and showed goo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A62D3/02C09K8/582B01F17/00C12R1/01A62D101/20C09K23/00
CPCA62D3/02C09K8/582A62D2101/20C12R2001/01C12N1/205C09K23/00
Inventor 齐义彬崔庆锋马原栋修建龙董汉平俞理黄立信
Owner PETROCHINA CO LTD
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