KR101636852B1 - Method And Agent For Soil Purification By Petroleum-Decomposing Microbe - Google Patents

Abstract

The soil contaminated with petroleum was classified as Arthrobacter sp. CO1 (Accession No. KCTC 12547BP), Methylobacterium fujisawaense CO2 (Accession No. KCTC 12548BP), Cowlobacter Henry Acinetobacter oleivorans CO4 (Accession No. KCTC 12550BP), Pseudomonas frederiksbergensis CO5 (accession number KCTC 12549BP), Accession No. KCTC Wherein the soil is purified by treatment with a petroleum-degrading microorganism strain selected from the group consisting of Pseudomonas sp. CO6 (Accession No. KCTC 12552 BP) and Pseudomonas sp. . When the soil is purified using the microbial strain according to the present invention, the soil is contaminated with petroleum at a low concentration of about 500 to 7000 ppm. In particular, as a purified soil primarily purified by physical or chemical methods, It is possible to purify contaminated soil effectively at a low concentration of ppm, so that the ecological integrity of the soil can be restored and the possibility of recycling can be increased.

Description

TECHNICAL FIELD The present invention relates to a petroleum-decomposing microbe,

The present invention relates to a soil remediation method and a soil remediation agent using petroleum-decomposing microorganisms, and more particularly, to a soil remediation method using a petroleum-decomposing microorganism suitable for purifying soil contaminated with oil at a concentration of about 500 to 7000 ppm, .

Existing methods used to purify soils contaminated with petroleum such as crude oil, gasoline, kerosene, and diesel that are not refined are mostly accompanied by deterioration of the ecological function of the soil during the purification process, due to strong physical and chemical reaction mechanisms . Because of this, purified soils are less adaptive to changes in external stresses such as sustainable production and climate change, and may require more time to restore the ecological function of the soil.

For this reason, most of the cases of reclamation are landfill or disposal without effective recycling. Since disposal sites such as construction materials that require soil resources, embankment sites, embankment construction, and landfill construction are constantly present, disposing of such sites as wastes rather than resources is a waste of resources do. To recycle these purified soils, it is necessary to improve soil quality to restore ecological function.

In order to ecologically recycle these purified soils, there are measures such as incorporation of a remedial agent to improve the deteriorated ecological function, or reinforcement of the grain size. However, due to the nature of soil purification that reduces pollutants within the range of purification standards according to land use methods, the pollution components remaining in the purified soil, especially the residual oil pollution components, have the greatest impact on ecological recycling. Removal of these residual petroleum pollutants will not have a significant impact on plant germination, but it will hinder plant growth in the future, so it is necessary to remove residual petroleum pollutants in order to ecologically recycle the cleaned soil.

Patent Registration No. 10-0435231 (registered on May 31, 2004) is a new middle / low temperature oilseed bacterial strain, YHLT-2KCTC 10203BP strain and a biological oil-removing method using the same. Japanese Patent Application Laid-Open No. 10-2004-0014714 (published Feb. 18, 2004) discloses an Asnithobacter sp. Strain having a lubricant-decomposing activity. Patent Registration No. 10-0676929 (registered on Jan. 25, 2007) discloses a method for removing Pseudomonas JN4 strain having lubricating oil resolution and contaminated lubricant from contaminated soil and ground water using the same. Patent Registration No. 10-1432425 (registered on Apr. 13, 2014) discloses an organic substance cleaning agent containing microorganisms in a plurality of strains identified from coastal tidal flats and waders.

The present invention is based on the fact that existing purified soil is used as a filler material or treated with waste and in order to compensate the disadvantage that the ecological function is lost in the purification process, The purpose of the present invention is to develop a petroleum-degrading microorganism capable of reducing the residual petroleum content, thereby recovering the ecological integrity of the purified soil and recycling it.

That is, the object of the present invention is to provide a soot soil contaminated with petroleum at a concentration of about 500 to 7000 ppm, especially a purified soil primarily purified by physical or chemical methods, The present invention provides a soil remediation method and a soil remediation agent using a petroleum-degrading microorganism suitable for purifying soil.

In order to accomplish the above object, the present invention provides a method for purifying soil using petroleum-decomposing microorganisms, wherein the soil contaminated with petroleum is treated with Arthrobacter sp. CO1 (Accession No. KCTC 12547BP) (Methylobacterium fujisawaense CO2) (Accession No. KCTC 12548BP), Caulobacter henricii CO3 (Accession No. KCTC 12549BP), Acinetobacter oleivorans CO4 (Accession No. KCTC Treated with a petroleum-degrading microorganism strain selected from the group consisting of Pseudomonas frederiksbergensis CO5 (Accession No. KCTC 12551 BP), Pseudomonas sp. CO6 (Accession No. KCTC 12552 BP) To purify the soil.

In the above method, it is preferable to further supply a nitrogen source and a phosphoric acid source to the soil so that the C / N / P ratio of the oil-contaminated soil is in the range of 100/10 / 1 to 100/1 / 0.5.

In addition, the petroleum content in the petroleum-contaminated soil is preferably in the range of 500 to 7000 ppm. Particularly, the soil contaminated with petroleum is preferably a purified soil primarily purified by physical or chemical methods, and it is preferable that the soil contains petroleum components in the range of 500 to 2000 ppm.

The present invention also relates to a process for the preparation of an antimicrobial composition comprising Arthrobacter sp. CO1 (Accession No. KCTC 12547BP), Methylobacterium fujisawaense CO2 (Accession No. KCTC 12548BP) (Accession No. KCTC 12551BP), Acinetobacter oleivorans CO4 (Accession No. KCTC 12550BP), Pseudomonas frederiksbergensis CO5 (Accession No. KCTC 12551BP), Caulobacter henricii CO3 (Accession No. KCTC 12549BP) , And Pseudomonas sp. CO6 (Accession No. KCTC 12552BP). The present invention also provides a soil remover comprising as an active ingredient, a petroleum-degrading microorganism strain selected from the group consisting of Pseudomonas sp.

The present invention also relates to a process for the preparation of an antimicrobial composition comprising Arthrobacter sp. CO1 (Accession No. KCTC 12547BP), Methylobacterium fujisawaense CO2 (Accession No. KCTC 12548BP) (Accession No. KCTC 12551BP), Acinetobacter oleivorans CO4 (Accession No. KCTC 12550BP), Pseudomonas frederiksbergensis CO5 (Accession No. KCTC 12551BP), Caulobacter henricii CO3 (Accession No. KCTC 12549BP) , And Pseudomonas sp. CO6 (Accession No. KCTC 12552BP). The present invention also provides a petroleum-degrading microorganism strain selected from the group consisting of Pseudomonas sp.

When the soil is purified using the microbial strain according to the present invention, the soil is contaminated with petroleum at a concentration of about 500 to 7000 ppm, in particular, a purified soil primarily purified by physical or chemical methods, So that it is possible to recover the ecological integrity of soil and increase the possibility of recycling.

Figure 1 shows the nucleotide sequence of Arthrobacter sp.
Figure 2 is a graph fujisawaense . < / RTI >
Figure 3 is a graph < / RTI > henricii .
Figure 4 shows that Acinetobacter oleivorans . < / RTI >
FIG. 5 shows the Pseudomonas frederiksbergensis . < / RTI >
Figure 6 shows the nucleotide sequence of Pseudomonas sp.
FIG. 7 is a graph showing a correlation between the concentration of high concentration TPH and the days of restoration of contaminated soil according to the microbial strain treatment according to an embodiment of the present invention.
FIG. 8 is a graph showing a correlation between the concentration of low concentration TPH and the days of restoration of contaminated soil according to the microbial strain treatment according to an embodiment of the present invention.

Soils that are heavily contaminated with petroleum, such as crude oil, gasoline, kerosene, light oil that are not normally refined, are purified by physical or chemical methods. These purified soils are low in concentrations of 500 to 2000 ppm, but still contain petroleum, which can not be recycled ecologically due to the residual oil.

SUMMARY OF THE INVENTION The present invention is intended to solve such problems of the prior art. That is, the present invention relates to a soil which is contaminated with petroleum components in the range of 500 to 7000 ppm, in particular as a purified soil primarily purified by physical or chemical methods, A purification agent, and a microorganism strain.

The present invention separates and identifies microbial strains to achieve the above object. Specifically, the microbial strains of the present invention are those obtained by selecting microbial strains having a high growth rate in a medium in which crude oil exists as a sole carbon source among microbial strains isolated from soil contaminated with crude oil. , Respectively.

First, a sample collected from a soil contaminated with crude oil was inoculated into a nutrient rich medium except for a carbon source, and the crude oil was further added to the medium so as to have a crude oil of 500 ppm as a carbon source, and cultured to grow crude oil-degrading microorganism strains.

After 10 days, 10 kinds of microorganism strains (M1 to M10) growing on a solid phase plate medium containing crude oil as the carbon source among the microorganism strains which occupied the dominant position in the culture medium were separated and 500 ppm of crude oil Were identified as CO1, CO2, CO3, CO4, CO5, and CO6, respectively.

One end of the selected microbial strain identified by using the partial nucleotide sequence determination and similarity analysis of the 16S rDNA bar, respectively, Arthrobacter sp., Methylobacterium fujisawaense, Caulobacter henricii , Acinetobacter Oleivorans , Pseudomonas frederiksbergensis , Pseudomonas sp. Of the isolates were Arthrobacter sp. CO1, Methylobacterium fujisawaense CO2, Caulobacter henricii CO3, Acinetobacter oleivorans CO4, Pseudomonas frederiksbergensis CO5, Pseudomonas sp. KCTC 12548BP, KCTC 12550BP, KCTC 12551BP, KCTC 12552BP and KCTC 12552BP deposited on February 5, 2014 with the accession number KCTC 12547BP, KCTC 12548BP, KCTC 12550BP, and KCTC 12552BP to the international depositary,

The isolated strains were found to be able to grow to OD = 10 or higher in both the essential minerals and the minimal medium containing crude oil contaminants as carbon source.

The strains according to the present invention not only have resistance to crude oil but also can be grown as a sole carbon source, so that the biological treatment of soil contaminated with crude oil was possible using these strains.

At this time, the strains of the above species can be used independently, and two or more of the six strains may be used in a mixed cultured state if necessary.

Also, in order to maximize the oil resolution of the strains, the nitrogen source and the phosphoric acid source necessary for decomposing the crude oil components of the strains were compared with the molar ratio of the contaminated carbon source, and ammonium sulphate, urea, ammonium nitrate and the like were added as the nitrogen source And the ratio of C / N / P is in the range of 100/10/1 to 100/1 / P, and the ratio of C / N / P of the soil is within the reference range, by supplying potassium phosphate, diphosphate, Lt; RTI ID = 0.0 > 0.5. ≪ / RTI >

In addition, the artificial crude oil decomposition effect using the strains and nutrients of the present invention developed as described above, and the natural crude oil decomposition effect in which the crude oil component is naturally volatilized and the crude oil component is decomposed by the natural microorganisms in the contaminated soil The present invention has been completed.

In the above, the microbial strains of the present invention were separated from soil polluted with crude oil and described that the soil contaminated with crude oil was purified. However, it is also possible to use a purified petroleum component instead of crude oil such as petroleum, kerosene, It is clear that it could also be used to purify the soil.

The above and other objects, features, and advantages of the present invention will become more apparent from the following detailed description of the present invention when taken in conjunction with the accompanying drawings.

Example

Example  1: Isolation of crude oil-degrading strain

For the isolation and identification of the crude oil-degrading strains, the medium components as shown in the following Table 1 were added and added in an amount of 10 g per liter of the medium contaminated with crude oil, followed by culturing at 160 rpm in a shaking incubator at 30 ° C.

Table 1

The culture broth grown in the proliferation step under the above conditions was applied to the crude degrading strain isolation medium as shown in Table 2, and cultured for 48 hours at 30 ° C in an incubator. Ten microorganism communities in which cell growth was confirmed by visual inspection were selected, ~ M10.

Table 2

Then, 10 strains selected from liquid nutrient medium containing crude oil as a carbon source were cultured for 48 hours, and the absorbance (OD) was measured at 600 nm. As shown in Table 3, 6 strains (CO1, CO2, CO3, CO4, CO5, CO6) were selected.

Table 3

As a result of partial sequencing of the 16S rDNAs selected as described above, the CO1 strain was Arthrobacter sp., The CO2 strain was Methylobacterium fujisawaense , CO3 strain Caulobacter henricii , CO4 strain is Acinetobacter oleivorans , CO5 strain is Pseudomonas frederiksbergensis , and CO6 strains were found to be novel strains corresponding to Pseudomonas sp.

Example 2: Nutrient supply

In order to maximize the degradation efficiency of the microbial strains, nitrogen and phosphorus are required as nutrients for the carbon sources contained in the pollutants. The concentration of phosphoric acid in the nitrate nitrogen and ammonia nitrogen available for the microorganisms in the contaminated soil is analyzed Ammonium sulfate, urea, ammonium nitrate, etc. as a nitrogen source so that the molar ratio of the carbon concentration to the carbon concentration to be decomposed in the contaminated TPH may be in the range of 100/10 / 1 to 100/1 / 0.5, potassium phosphate, potassium phosphate diphosphate . In the examples of the present invention, C / N / P = 100/10/1 was prepared as shown in the following Table 4 and added to the soil together with the new microorganism of Example 1.

Table 4

Example 3: Restoration of contaminated soil of crude oil

A vinyl house treatment plant having a size of 15 m x 90.4 m was prepared using the strains and nutrients of the present invention of Examples 1 and 2, sand, non-woven fabric, and collecting molds for collecting leachate were installed on the lower portion. .

After adding 3 ml of the strain of the present invention cultured at 110 ⁹ CFU / ml per ton of the contaminated soil, the number of moles of carbon contaminated crude oil component and the concentration of nitrogen and phosphoric acid in the contaminated crude oil component were measured as in Example 2, and the C / N / P concentration Was prepared three days before the strain injection. The contaminated soil was inverted 23 times per week by using an excavator, and the water was supplied to the spraying device Were sprayed every day.

As a result of confirming the initial and restoration contamination concentrations contaminated with crude oil by gravimetric method, it was confirmed that after about 50 days, the concentration was restored from approximately 7,000 ppm to approximately 1,000 ppm, and it was confirmed that after approximately 50 days, approximately 250 ppm was recovered from approximately 820 ppm. These are shown in FIGS. 7 and 8. FIG. 7 shows the correlation between the high concentration TPH concentration (7000 ppm) and the days of restoration of the contaminated soil according to the microbial strain treatment of the present invention, and FIG. 8 shows the correlation between the low concentration TPH concentration (800 ppm or higher) Respectively.

Biotechnology Institute Gene Bank KCTC12547BP 20140205 Biotechnology Institute Gene Bank KCTC12547BP 20140205 Biotechnology Institute Gene Bank KCTC12549BP 20140205 Biotechnology Institute Gene Bank KCTC12550BP 20140205 Biotechnology Institute Gene Bank KCTC12551BP 20140205 Biotechnology Institute Gene Bank KCTC12552BP 20140205

<110> OIKOS CO., LTD. <120> Method And Agent For Soil Purification By Petroleum-Decomposing          Microbe <130> kyd140076 <160> 6 <170> Kopatentin 2.0 <210> 1 <211> 891 <212> DNA <213> Arthrobacter sp. <400> 1 ttagtggcga acgggtgagt aacacgtgag taacctgccc ttgactctgg gataagcctg 60 ggaaactggg tctaataccg gatatgactc ctcatcgcat ggtggggggt ggaaagcttt 120 tgtggttttg gatggactcg cggcctatca gcttgttggt gaggtaatgg ctcaccaagg 180 cgacgacggg tagccggcct gagagggtga ccggccacac tgggactgag acacggccca 240 gactcctacg ggaggcagca gtggggaata ttgcacaatg ggcgcaagcc tgatgcagcg 300 acgccgcgtg agggatgacg gccttcgggt tgtaaacctc tttcagtagg gaagaagcga 360 aagtgacggt acctgcagaa gaagcgccgg ctaactacgt gccagcagcc gcggtaatac 420 gtagggcgca agcgttatcc ggaattattg ggcgtaaaga gctcgtaggc ggtttgtcgc 480 gtctgccgtg aaagtccggg gctcaactcc ggatctgcgg tgggtacggg cagactagag 540 tgatgtaggg gagactggaa ttcctggtgt agcggtgaaa tgcgcagata tcaggaggaa 600 caccgatggc gaaggcaggt ctctgggcat taactgacgc tgaggagcga aagcatgggg 660 agcgaacagg attagatacc ctggtagtcc atgccgtaaa cgttgggcac taggtgtggg 720 ggacattcca cgttttccgc gccgtagcta acgcattaag tgccccgcct ggggagtacg 780 gccgcaaggc taaaactcaa aggaattgac gggggcccgc acaagcggcg gagcatgcgg 840 attaattcga tgcaacgcga agaaccttac caaggcttga catggaccgg a 891 <210> 2 <211> 873 <212> DNA <213> Methylobacterium fujisawaense <400> 2 taacgcgtgg gaacgtgcct tccggttcgg aataaccctg ggaaactagg gctaataccg 60 gatacgccct tatggggaaa ggtttactgc cggaagatcg gcccgcgtct gattagctag 120 ttggtggggt aacggcctac caaggcgacg atcagtagct ggtctgagag gatgatcagc 180 cacactggga ctgagacacg gcccagactc ctacgggagg cagcagtggg gaatattgga 240 caaggggcgc aagcctgatc cagccatgcc gcgtgagtga tgaaggcctt agggttgtaa 300 agctctttta tccgggacga taatgacggt accggaggaa taagccccgg ctaacttcgt 360 gccagcagcc gcggtaatac gaagggggct agcgttgctc ggaatcactg ggcgtaaagg 420 gcgcgtaggc ggcgttttaa gtcgggggtg aaagcctgtg gctcaaccac agaatggcct 480 tcgatactgg gacgcttgag tatggtagag gttggtggaa ctgcgagtgt agaggtgaaa 540 ttcgtagata ttcgcaagaa caccggtggc gaaggcggcc aactggacca ttactgacgc 600 tgaggcgcga aagcgtgggg agcaaacagg attagatacc ctggtagtcc acgccgtaaa 660 cgatgaatgc cagctgttgg ggtgcttgca ccgcagtagc gcagctaacg ctttgagcat 720 tccgcctggg gagtacggtc gcaagattaa aactcaaagg aattgacggg ggcccgcaca 780 agcggtggag catgtggttt aattcgaagc aacgcgcaga accttaccat cctttgacat 840 ggcgtgttac ccagagagat ttggggtcca ctt 873 <210> 3 <211> 873 <212> DNA <213> Caulobacter henricii <400> 3 tcgaacggat ccttcgggat tagtggcgga cgggtgagta acacgtggga acgtgccctt 60 tggttcggaa caactcaggg aaacttgagc taataccgga tgtgcccttc gggggaaaga 120 tttatcgcca ttggagcggc ccgcgtagga ttagctagtt ggtggggtaa aggcccacca 180 aggcgacgat ccttagctgg tctgagagga tgatcagcca cattgggact gagacacggc 240 ccaaactcct acgggaggca gcagtgggga atcttgcgca atgggcgaaa gcctgacgca 300 gccatgccgc gtgaatgatg aaggtcttag gattgtaaaa ttctttcacc ggggacgata 360 atgacggtac ccggagaaga agccccggct aacttcgtgc cagcagccgc ggtaatacga 420 agggggctag cgttgctcgg aattactggg cgtaaaggga gcgtaggcgg actgtttagt 480 cagaggtgaa agcccagggc tcaaccttgg aatagccttt gatactggca gtcttgagta 540 cggaagaggt atgtggaact ccgagtgtag aggtgaaatt cgtagatatt cggaagaaca 600 ccagtggcga aggcgacata ctggtccgtt actgacgctg aggctcgaaa gcgtggggag 660 caaacaggat tagataccct ggtagtccac gctgtaaacg atgagtgcta gttgtcggca 720 ggcatgcctg tcggtgacgc agctaacgca ttaagcactc cgcctgggga gtacggtcgc 780 aagattaaaa ctcaaaggaa ttgacggggg cccgcacaag cggtggagca tgtggtttaa 840 ttcgaagcaa cgcgcagaac cttaccacct ttt 873 <210> 4 <211> 869 <212> DNA <213> Acinetobacter oleivorans <400> 4 tgcaagtcga gcggagagag gtagcttgct actgatctta gcggcggacg ggtgagtaat 60 gcttaggaat ctgcctatta gtgggggaca acatttcgaa aggaatgcta ataccgcata 120 cgtcctacgg gagaaagcag gggatcttcg gaccttgcgc taatagatga gcctaagtcg 180 gattagctag ttggtggggt aaaggcctac caaggcgacg atctgtagcg ggtctgagag 240 gatgatccgc cacactggga ctgagacacg gcccagactc ctacgggagg cagcagtggg 300 gaatattgga caatgggcgg aagcctgatc cagccatgcc gcgtgtgtga agaaggcctt 360 atggttgtaa agcactttaa gcgaggagga ggctacttta gttaatacct agagatagtg 420 gacgttactc gcagaataag caccggctaa ctctgtgcca gcagccgcgg taatacagag 480 ggtgcaagcg ttaatcggat ttactgggcg taaagcgcgc gtaggcggct aattaagtca 540 aatgtgaaat ccccgagctt aacttgggaa ttgcattcga tactggttag ctagagtgtg 600 ggagaggatg gtagaattcc aggtgtagcg gtgaaatgcg tagagatctg gaggaatacc 660 gatggcgaag gcagccatct ggcctaacac tgacgctgag gtgcgaaagc atggggagca 720 aacaggatta gataccctgg tagtccatgc cgtaaacgat gtctactagc cgttggggcc 780 tttgaggctt tagtggcgca gctaacgcga taagtagacc gcctggggag tacggtcgca 840 agactaaaac tcaaatgaat tgacggggg 869 <210> 5 <211> 953 <212> DNA <213> Pseudomonas frederiksbergensis <400> 5 tgcaagtcga gcggcagcac gggtacttgt acctggtggc gagcggcgga cgggtgagta 60 atgcctagga atctgcctgg tagtggggga taacgttcgg aaacgaacgc taataccgca 120 tacgtcctac gggagaaagc aggggacctt cgggccttgc gctatcagat gagcctaggt 180 cggattagct agttggtgag gtaatggctc accaaggcga cgatccgtaa ctggtctgag 240 aggatgatca gtcacactgg aactgagaca cggtccagac tcctacggga ggcagcagtg 300 gggaatattg gacaatgggc gaaagcctga tccagccatg ccgcgtgtgt gaagaaggtc 360 ttcggattgt aaagcacttt aagttgggag gaagggcagt tacctaatac gtaattgttt 420 tgacgttacc gacagaataa gcaccggcta actctgtgcc agcagccgcg gtaatacaga 480 gggtgcaagc gttaatcgga attactgggc gtaaagcgcg cgtaggtggt tcgttaagtt 540 ggatgtgaaa tccccgggct caacctggga actgcattca aaactgtcga gctagagtat 600 ggtagagggt ggtggaattt cctgtgtagc ggtgaaatgc gtagatatag gaaggaacac 660 cgtggcgaa ggcgaccacc tggactgata ctgacactga ggtgcgaaag cgtggggagc 720 aaacaggatt agataccctg gtagtccacg ccgtaaacga tgtcaactag ccgttgggag 780 ccttgagctc ttagtggcgc agctaacgca ttaagttgac cgcctgggga gtacggccgc 840 aaggttaaaa ctcaaatgaa ttgacggggg cccgcacaag cggtggagca tgtggtttaa 900 ttcgaagcaa cgcgaagaac cttaccaggc cttgacatcc aatgaacttt cca 953 <210> 6 <211> 858 <212> DNA <213> Pseudomonas sp. <400> 6 tgcaagtcga gcggatgaaa ggagcttgct cctggattca gcggcggacg ggtgagtaat 60 gcctaggaat ctgcctggta gtgggggaca acgtttcgaa aggaacgcta ataccgcata 120 cgtcctacgg gagaaagcag gggaccttcg ggccttgcgc tatcagatga gcctaggtcg 180 gattagctag ttggtgaggt aatggctcac caaggcgacg atccgtaact ggtctgagag 240 gatgatcagt cacactggaa ctgagacacg gtccagactc ctacgggagg cagcagtggg 300 gaatattgga caatgggcga aagcctgatc cagccatgcc gcgtgtgtga agaaggtctt 360 cggattgtaa agcactttaa gttgggagga agggttgtag attaatactc tgcaattttg 420 acgttaccga cagaataagc accggctaac tctgtgccag cagccgcggt aatacagagg 480 gtgcaagcgt taatcggaat tactgggcgt aaagcgcgcg taggtggttc gttaagttgg 540 atgtgaaatc cccgggctca acctgggaac tgcatccaaa actggcgagc tagagtatgg 600 tagagggtgg tggaatttcc tgtgtagcgg tgaaatgcgt agatatagga aggaacacca 660 gtggcgaagg cgaccacctg gactgatact gacactgagg tgcgaaagcg tggggagcaa 720 acaggattag ataccctggt agtccacgcc gtaaacgatg tcaactagcc gttgggagcc 780 ttgagctctt agtggcgcag ctaacgcatt aagttgaccg cctggggagt acggccgcaa 840 ggttaaaact caaatgaa 858

Claims (6)

The soil contaminated with petroleum was classified as Arthrobacter sp. CO1 (Accession No. KCTC 12547BP), Methylobacterium fujisawaense CO2 (Accession No. KCTC 12548BP), Cowlobacter Henry Acinetobacter oleivorans CO4 (Accession No. KCTC 12550BP), Pseudomonas frederiksbergensis CO5 (accession number KCTC 12549BP), Accession No. KCTC Wherein the soil is purified by treatment with a petroleum-degrading microbial strain selected from the group consisting of Pseudomonas sp. CO6 (Accession No. KCTC 12552BP). The method according to claim 1,
Wherein the nitrogen source and the phosphoric acid source are further supplied to the soil so that the C / N / P ratio of the soil contaminated with the petroleum is in the range of 100/10 / 1 to 100/1 / 0.5. Soil remediation method. 3. The method according to claim 1 or 2,
Wherein the petroleum component in the petroleum-contaminated soil is in the range of 500 to 7000 ppm. 3. The method according to claim 1 or 2,
Wherein the soil contaminated with petroleum is a purified soil primarily purified by a physical or chemical method and comprises a petroleum component in a range of 500 to 2000 ppm. (Arthrobacter sp. CO1) (Accession No. KCTC 12547BP), Methylobacterium fujisawaense CO2 (KCTC 12548BP), Caulobacter henricii CO3 KCTC 12549BP), Acinetobacter oleivorans CO4 (KCTC 12550BP), Pseudomonas frederiksbergensis CO5 (KCTC 12551BP), Pseudomonas sp. CO6 (KCTC 12551BP), Pseudomonas sp. 12552BP) as an active ingredient. (Arthrobacter sp. CO1) (Accession No. KCTC 12547BP), Methylobacterium fujisawaense CO2 (KCTC 12548BP), Caulobacter henricii CO3 KCTC 12549BP), Acinetobacter oleivorans CO4 (KCTC 12550BP), Pseudomonas frederiksbergensis CO5 (KCTC 12551BP), Pseudomonas sp. CO6 (KCTC 12551BP), Pseudomonas sp. 12552BP). &Lt; / RTI &gt;

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