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Rhodococcus erythropolis and application thereof

A technology of Rhodococcus erythropolis and bacteria agent, which is applied in the field of environmental microorganisms, can solve the problems of low degradation ability of microbial petroleum hydrocarbons, and achieve the effects of prolonging residence time, increasing survival rate, and improving solubility

Inactive Publication Date: 2015-11-04
TIANJIN YISHI KAIYUAN TECH DEV CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims at the technical defects of the prior art, provides a kind of Rhodococcus erythropolis and its application, so as to solve the technical problem of the prior art that the degradation ability of microbial petroleum hydrocarbons is low

Method used

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  • Rhodococcus erythropolis and application thereof
  • Rhodococcus erythropolis and application thereof
  • Rhodococcus erythropolis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment l

[0051] Example 1: Screening of Rhodococcus erythoropolis HL-6 strain provided by the present invention.

[0052] Take 1 g of oil-contaminated soil from Xinjiang Oilfield and put it in 100 mL of inorganic salt medium with 0.5% diesel oil as the sole carbon source, and place it in a constant temperature shaker at 200 r / min at 25 ° C for 72 h for enrichment. Take 2mL of the enriched bacteria liquid and transfer it to a new 100mL inorganic salt medium, and the culture method is the same as above. After three cycles of enrichment, the main strain HL-6 was finally obtained. Take one ring of HL-6 and put it into a test tube filled with 5mL LB culture solution, culture it with shaking at 25°C for 48h, and use it as the seed solution. Put 2% inoculum into a 100mL Erlenmeyer flask equipped with 30mL inorganic salt medium, incubate with shaking at 25°C for 72h, extract with n-hexane, and detect the degradation of diesel oil by gas chromatography (GC). And by measuring the emulsificatio...

Embodiment 2

[0056] Example 2: Morphological and physiological and biochemical characteristics of the Rhodococcus erythoropolis HL-6 strain provided by the present invention.

[0057] Refer to the experimental method of "Bergey's Manual of Systematic Bacteriology" (Vol.Ⅷ) to detect its Gram staining, cell size and shape, presence or absence of flagella and spores, growth temperature, growth pH range, and NaCI tolerance. Catalase, nitrate reduction, urease, escin hydrolysis, 2,3-butanediol, aspartic acid, oxidase, M.R., V-P, indole, nitrite protogenesis, denitrification, gelatin liquefaction, starch Hydrolysis, adenine hydrolysis experiment and the use of mannitol, inositol experiment.

[0058] The results showed that the strain was Gram-positive, and the strain was short rod-shaped, without flagella, immobile, aerobic, and did not produce spores. The cell size was 2.0-3.0 μm (length) × 0.8-1 μm (width), with a typical figure-of-eight arrangement. The growth temperature is 10-37°C, the gr...

Embodiment 3

[0059] Example 3: PCR amplification and sequence determination of the 16S rRNA gene of the Rhodococcus erythoropolis HL-6 strain provided by the present invention.

[0060] Inoculate the strain in LB medium, culture it on a shaker at 25°C (200rpm) for 10 hours, collect the bacteria by centrifugation, resuspend, add lysozyme and SDS to break the wall, extract genomic DNA by the phenol-chloroform method, and use the normal phase primer 27F (5'-GAGAGTTTGATCCTGGCTCAG-3') reverse primer 1541R (5'-AAGGAGGTGATCCAGCC-3'), use this pair of primers to perform PCR amplification of its 16S rDNA tomb, and send the amplification primers to Beijing Aoke 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 16S rRNA gene sequence is 1470bp, the accession number in GenBank is JQ839141, and the similarity value of Rhodococcus erythropolis EPWF 16S rDNA (GenBank accession No. AY822047) is the highest, wh...

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Abstract

The invention provides efficient hydrocarbon-degradation rhodococcus erythropolis HL-6 and application thereof. The rhodococcus is obtained from petroleum polluted soil in a Xinjiang petroleum field through separation, wherein the growth temperature is 10-37 DEG C, the optimal temperature is 15-25 DEG C, the growth pH ranges from 5 to 10, and the sodium chloride tolerability is 0-5%. Alkane or diesel oil is used as a unique carbon source for growth. The rhodococcus erythropolis has the high petroleum hydrocarbon degradation capability, can degrade C11-C36 n-alkanes and diesel oil, has the good tolerance for diesel oil, can still grow under 30% diesel oil, OD600 can reach 1.25, and the emulsifying activity is 44%. 0.5% diesel oil is added, and then the degradation rate reaches 78.5% after 72h. A bacterial strain can produce biosurfactant in a hydrophilic substrate and a hydrophobic substrate, and has the good emulsification effect on saturated alkane and aromatic compounds.

Description

technical field [0001] The invention relates to the technical field of environmental microorganisms, in particular to a Rhodococcus erythropolis and an application thereof. Background technique [0002] With the frequent occurrence of offshore oil development and tanker accidents, offshore oil pollution is becoming more and more serious, which directly affects the development of marine aquaculture and fishing industries. After an oil spill accident, part of the oil spilled on the surface can be removed by physical methods (such as mechanical recovery, etc.), and the remaining oil spilled will drift to the tidal flat under the action of wind, waves, and currents. The refractory aromatic components (carcinogens) in petroleum remain in the tidal flat soil environment for a long time, causing huge and lasting damage to the tidal flat ecosystem and human health. Microbial treatment technology is an internationally recognized new technology for efficient repair of oil pollution. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N11/14B09C1/10
Inventor 黄磊李申
Owner TIANJIN YISHI KAIYUAN TECH DEV CO LTD
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