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Rhodococcus ruber and application thereof in degradation of hydrocarbon compounds

A technology for Rhodococcus rhodochrous and chemical compounds is applied in the fields of microbial biotechnology and environmental biology, and can solve the problems of the degradation of less hydrocarbon compounds and the great environmental harm.

Inactive Publication Date: 2011-11-16
SHANTOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among oil pollutants, hydrocarbons are the most harmful to the environment. At present, there are few literature reports on the degradation of hydrocarbons, especially high molecular weight hydrocarbons, by Rhodococcus

Method used

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  • Rhodococcus ruber and application thereof in degradation of hydrocarbon compounds
  • Rhodococcus ruber and application thereof in degradation of hydrocarbon compounds
  • Rhodococcus ruber and application thereof in degradation of hydrocarbon compounds

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1: Isolation and screening of bacterial strain Rhodococcus sp.CGMCC NO.2343 provided by the present invention

[0059] The sample comes from the surface sediments in the sea area of ​​Xiamen. Weigh 5g of seabed silt, add 50ml of sterile water to shake for 2h, let it stand for 30min, absorb the clear liquid from the upper layer, and transfer it to the enrichment medium, that is, the medium of inorganic salts plus polycyclic aromatic hydrocarbons. Composition: (NH 4 ) 2 SO 4 , 1.0g; Na 2 HPO 4 , 0.8g; KH 2 PO 4 , 0.2g; MgSO 4 ·7H 2 O, 0.2g; CaCl 2 2H 2 O, 0.1g; FeCl 3 ·6H 2 O, 5 mg; (NH 4 ) 6 Mo 7 o 24 4H 2 O, 1mg; distilled water 1000ml; polycyclic aromatic hydrocarbons 50mg; pH 7.0, sterilized at 121°C for 30min), after culturing for 15 days, transfer 1ml to culture medium containing 50mg / l polycyclic aromatic hydrocarbon inorganic salts for one week, then transfer to fresh 50mg / l polycyclic aromatic hydrocarbon inorganic salt culture solution. ...

Embodiment 2

[0061]Embodiment 2: PCR amplification and sequence determination of the 16S rDNA gene of bacterial strain Rhodococcus sp.CGMCC NO.2343 provided by the present invention

[0062] The colony PCR method was adopted, and the single colony lysate was directly taken as a template for PCR. The PCR primers for amplifying 16S rDNA were a pair of universal primers, 27f: 5’-AGAGTTTGATCCTGGCTCAG-3’, 1492R: 5’-GGTTACCTTGTTACGACT-3’. The PCR reaction conditions were: 94°C for 3min; 30 cycles of 94°C for 30s, 55°C for 30s, and 72°C for 2min; 72°C for 10min. The PCR amplified product was purified and connected to T vector (Shanghai Sangon TA cloning kit), and sent to Shanghai Yingjun Biotechnology Co., Ltd. for sequencing. Blast analysis was performed on the sequencing results and the sequences on Genbank. The length of the sequence is about 1500bp, and the accession number in Genbank is EF634456. The strain of the present invention has great homology with Rhodococcus erythrococcus.

Embodiment 3

[0063] Embodiment 3: the bacterial strain Rhodococcus sp.CGMCC NO.2343 provided by the present invention floats in the characteristic of oily substance

[0064] Cultivate Rhodococcus sp.CGMCC NO.2343 to OD in 2216E medium and be 0.8, collect thalli, inoculate in the 50ml inorganic salt culture fluid containing 4% diesel oil or paraffin after washing with MSM (a batch of cultivation The solution is placed in a test tube with a length of 12 cm and a diameter of 4.5 cm, and another batch is placed in a 150 ml Erlenmeyer flask), and cultivated at 30° C. and 150 rpm for 10 days. Control the non-inoculated parallel samples, and observe the changes of the oily substances floating on the surface of the culture medium. For the test tube culture samples, draw 10ul of the culture solution of each layer and gradually dilute it to 10 -3 、10 -6 、10 -9 , Take 50μl of each dilution and spread on 2216E plate. Count the number of colonies after incubation at 30°C for 48 h.

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Abstract

The invention relates to a bacterial strain of Rhodococcus ruber P14 CGMCC NO.2343. The bacterial strain has the characteristic of floating up from oil matters; the bacterial strain can grow by taking the oil matters as unique carbon source and energy source and degrade the oil matters; and the bacterial strain can grow by taking polycyclic aromatic hydrocarbon as unique carbon source and energy source and degrade hydrocarbon compounds, such as phenanthrene, pyrene, benzopyrene, and the like. The bacterial strain can degrade the oil matters and hydrocarbon compounds, especially the polycyclicaromatic hydrocarbon can be applied in the biological treatment of oily waste water and the biological repair (biological remediation) of oil-contaminated soil.

Description

technical field [0001] The invention belongs to the field of microbial biotechnology and environmental biotechnology, and specifically relates to a strain of Rhodococcus sp., whose preservation number is: CGMCC NO.2343. Degrades oily substances containing hydrocarbons. Background technique [0002] With the development of the economy, the human demand for energy is also expanding. All countries have accelerated the development and utilization of oil and gas resources. From deserts to oceans, from uninhabited areas to densely populated areas, more and more oil and gas wells appear all over the world. . Petroleum is a complex mixture composed of thousands of substances with different chemical properties, mainly including saturated hydrocarbons, aromatic hydrocarbons, asphaltenes, resins, etc. A series of oil pollution problems are caused by pollution and omission accidents in the process of oil extraction, smelting, use, and transportation, as well as the discharge of oily w...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A62D3/02C02F3/34B09C1/10C12S13/00C02F103/36A62D101/20C12R1/01
Inventor 胡忠徐艳宋小慧伦镜盛黄通旺
Owner SHANTOU UNIV
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