Petroleum degrading bacterium, obtaining method and application thereof in crude oil degradation

A technology for oil degrading bacteria and crude oil, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve the problems of long-chain hydrocarbons that need to be improved, and achieve fast degradation, high degradation efficiency, and good degradation effect Effect

Active Publication Date: 2018-07-20
YONKER ENVIRONMENTAL PROTECTION
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Chinese patent CN 104017747 B discloses a oil-degrading bacterium in oily sludge and its application. Corynebacterium glutamicum is used to degrade oily sludge. It degrades at 37°C for 30 days. The degradation rate of saturated hydrocarbons is The degradation rate to total petroleum hydrocarbons is 20.74%, and the degradation rate to total petroleum hydrocarbons is 39.69%. The degrading bacteria were separated and screened out, and added to pollutants containing 0.5% crude oil. After 7 days of degradation, most of the short-chain hydrocarbons and medium- and long-chain hydrocarbons can be degraded, and the degradation rate of long-chain hydrocarbons can reach 70-80%. ; Chinese patent CN 102...

Method used

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  • Petroleum degrading bacterium, obtaining method and application thereof in crude oil degradation
  • Petroleum degrading bacterium, obtaining method and application thereof in crude oil degradation
  • Petroleum degrading bacterium, obtaining method and application thereof in crude oil degradation

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Effect test

Embodiment 1

[0037] Enrichment and Isolation of Mycobacterium Mycobacteium sp.CSC-6

[0038] (1) On-site collection of oil-contaminated soil near Shengli Oilfield, with crude oil as the only carbon source, to screen out bacteria that can efficiently degrade crude oil. Take by weighing 5g oil sludge sample, add and fill 50mL inorganic salt culture medium (1.00g / L NH 4 Cl, 0.05g / L CaCl 2 , 0.10g / L MgSO 4 .7H 2 O, 1.00g / L K 2 HPO 4 , 1.00g / L KH 2 PO 4 ) in an Erlenmeyer flask at 30°C and 120r / min on a shaker for 7 days. After the culture medium became turbid, pipette 5mL of the culture medium and transfer it into a fresh 50mL inorganic salt medium, and continuously transfer and enrich the culture for 6 times.

[0039] (2) Add 1mL of the culture solution obtained from 6 times of enrichment culture into 9mL of PBS solution, serially dilute it 6 times, and coat the plate with different dilution gradient culture solution, culture in a constant temperature incubator at 30°C for...

Embodiment 2

[0042] Degradation performance of mycobacterium Mycobacteium sp.CSC-6

[0043] Use LB medium to expand the culture of strain CSC-6, centrifuge and separate the bacteria after 48 hours, prepare a suspension with PBS and adjust the OD value to 1.0, and inoculate the bacteria solution to a crude oil concentration of 150 at a ratio of 2% by volume. mg / L inorganic salt medium, take the crude oil concentration of 150 mg / L inorganic salt medium that does not contain the strain CSC-6 as a blank, place it on a shaker at 30°C and 120r / min for shaking culture for 1, 3 days, and use n-Hexane extracts the remaining petroleum hydrocarbons. The experimental group and the blank group were analyzed respectively by GC-MS, and the results were as follows: figure 1 As shown, after 1 day of degradation, the degradation rate of C10-C16 in crude oil was 55.44%, the degradation rate of C17-C28 was 45.86%, and the degradation rate of total petroleum hydrocarbons was 48.27%; The degradation ra...

Embodiment 3

[0045] Use LB medium to expand the culture of strain CSC-6, centrifuge and separate the bacteria after 48 hours, and prepare a suspension with PBS and adjust the OD value to 1.0, and inoculate the bacteria solution to a crude oil concentration of 300 at a ratio of 2% by volume. mg / L inorganic salt medium, take the crude oil concentration of 300mg / L inorganic salt medium that does not contain the strain CSC-6 as a blank, place it on a shaker at 30°C and 120r / min for shaking culture for 1, 3 days, and use n-Hexane extracts the remaining petroleum hydrocarbons. The experimental group and the blank group were analyzed by GC-MS. After 1 day of degradation, the degradation rate of C10-C16 in crude oil was 23.95%, the degradation rate of C17-C28 was 70.47%, and the degradation rate of C29-C36 was 100%. The degradation rate of total petroleum hydrocarbons was 69.58%; after 3 days of degradation, the degradation rate of C10-C16 in crude oil was 100%, the degradation rate of C17-C...

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Abstract

The invention relates to a petroleum degrading bacterium and an application thereof in crude oil degradation. The petroleum degrading bacterium is named as Mycobacterium sp.CSC-6, and is a Gram-negative bacterium. The bacterial strain is preserved in China Center for Type Culture Collection (CCTCC) in Nov. 27th, 2017. The preservation number is CCTCC No. M2017726. The bacterial strain is in a rodshape, has a light yellow color, and is opaque. After 48 hour culture, the diameter of the bacterial colony is 9.0-11.0 mm. The log-in number of the 16Sr DNA of the bacterial strain in GeneBank is MG490974.1. Compared with the prior art, the provided bacterial strain has high degradation efficiency, can well degrade short chain hydrocarbons, middle chain hydrocarbons, and long chain hydrocarbons,and is capable of completely degrading middle chain hydrocarbons and long chain hydrocarbons in crude oil, and at the same time, the degradation speed is quick.

Description

technical field [0001] The invention relates to a petroleum degrading bacterium, its obtaining method and its application in degrading crude oil. Background technique [0002] As the blood of industry, petroleum is closely related to human life. However, during the process of mining, refining, storage, transportation, and use, petroleum is easy to enter the soil environment, causing changes in soil physical and chemical properties. For example: petroleum will block the pore structure of the soil to make it salinized, bituminized, compacted, and reduce the water permeability of the soil; the reactive groups rich in petroleum can combine with inorganic nitrogen and phosphorus in the soil and limit nitrification and Dephosphorylation reduces the available nitrogen and phosphorus content of the soil, leading to changes in the carbon-nitrogen and carbon-phosphorus ratios of soil organic matter, thereby reducing the types and quantities of microorganisms in the soil, and reducing...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02B09C1/10C12R1/32
CPCB09C1/10C12N1/02C12N1/20C12N1/205C12R2001/32
Inventor 罗启仕赵秀红崔长征闫淑兰周玉强缪周伟吴晴雯
Owner YONKER ENVIRONMENTAL PROTECTION
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