Method for decomposing hydrocarbon compound, method for purifying soil and / or water, composition for decomposing hydrocarbon compound, and microorganism capable of decomposing hydrocarbon compound

Abstract

To provide a technique for decomposing hydrocarbon compounds in soil and / or water, using microorganisms with hydrocarbon-decomposing capabilities even under anaerobic environments and low safety risks.SOLUTION: The present invention provides a method for decomposing hydrocarbon compounds in soil and / or water using microorganisms, the method comprising a step of contacting the soil and / or water containing the hydrocarbon compounds with the microorganisms. The microorganisms refer to one or more microorganisms belonging to any of the species of (1) Gordonia neofelifaecis, (2) Gordonia rubripertincta, and (3) Rhodococcus rhodochrous.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present disclosure relates to a method for decomposing hydrocarbon compounds, a method for purifying soil and / or water, a composition for decomposing hydrocarbon compounds, and a microorganism having an ability to decompose hydrocarbon compounds. More specifically, the present disclosure relates to a method for decomposing hydrocarbon compounds contained in soil and / or water using a microorganism. [Background technology]

[0002] When reusing the sites of factories, gas stations, etc., the soil at the sites may be contaminated with mineral oils and other chemicals, making it necessary to take measures against such contaminated soil. One method for purifying contaminated soil is bioremediation, which utilizes the ability of microorganisms to decompose pollutants. Bioremediation has the advantages of low environmental impact and low processing costs.

[0003] In the purification of contaminated soil, air is supplied near the ground surface, so purification is carried out in an aerobic state, but as the depth from the ground surface increases, the supply of air becomes less and the soil gradually becomes anaerobic. Therefore, in order to make the decomposition ability of microorganisms manifest even in soil deep below the ground surface, in the purification of contaminated soil, pipes that supply air underground are installed and the soil is stirred periodically, artificially creating aerobic conditions even in soil deep underground.

[0004] Patent Document 1 discloses a microorganism capable of decomposing hydrocarbon compounds or oils even under anaerobic conditions. The microorganism belongs to Pseudomonas aeruginosa and must be handled at Biosafety Level 2 (BSL2), raising safety concerns about its application to outdoor bioremediation. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] JP 2015-042612 A Summary of the Invention [Problem to be solved by the invention]

[0006] The main objective of the present disclosure is to provide a technology for decomposing hydrocarbon compounds contained in soil and / or water using microorganisms that exhibit the ability to decompose hydrocarbon compounds even under anaerobic conditions and have few safety concerns. [Means for solving the problem]

[0007] In order to solve the above problems, this disclosure provides the following [1]-

[22] . [1] A method for decomposing hydrocarbon compounds contained in soil and / or water by microorganisms, comprising the steps of: contacting the soil and / or the water containing the hydrocarbon compound with the microorganism, The method, wherein the microorganism is one or more microorganisms belonging to any of the following species (1) to (3): (1) Gordonia neofelifaecis (2) Gordonia rubripertincta (3) Rhodococcus rhodochrous [2] The microorganism is It is a microorganism of any of the following strains (1)-(7), or The method according to [1], wherein the microorganism has 16S rRNA comprising a base sequence having 90% or more sequence identity with the strain and exhibits the ability to decompose hydrocarbon compounds under anaerobic conditions. (1) Gordonia neoferrifaesis strain MCI526 (accession number: NITE P-03813) (2) Gordonia lubripeltinctula DSMZ43570 strain (3) Rhodococcus rhodocrus ATCC21291 strain (4) Rhodococcus rhodocrus ATCC14347 strain (5) Rhodococcus rhodocrus ATCC14348 strain (6) Rhodococcus rhodocrus ATCC14349 strain (7) Rhodococcus rhodocrus ATCC14350 strain [3] The method according to [1] or [2], wherein the step is carried out under anaerobic conditions. [4] The method according to any one of [1] to [3], wherein the hydrocarbon compound is a long-chain n-alkane of C12 to C44. [5] The method according to [4], wherein the hydrocarbon compound is a long-chain n-alkane of C28 to C44. [6] The method according to [4], wherein the hydrocarbon compound is octadecane.

[0008] [7] A method for remediating soil and / or water contaminated with hydrocarbon compounds, comprising: The method includes a step of contacting the soil and / or the water with microorganisms and decomposing the hydrocarbon compounds by the action of the microorganisms, The method, wherein the microorganism is one or more microorganisms belonging to any of the following species (1) to (3): (1) Gordonia neofelifaecis (2) Gordonia rubripertincta (3) Rhodococcus rhodochrous [8] The microorganism is It is a microorganism of any of the following strains (1)-(7), or The method according to [7], wherein the microorganism has 16S rRNA comprising a base sequence having 90% or more sequence identity with the strain and exhibits the ability to decompose hydrocarbon compounds under anaerobic conditions. (1) Gordonia neoferriphaesis strain MCI526 (accession number: NITE P-03813) (2) Gordonia lubripeltinctula DSMZ43570 strain (3) Rhodococcus rhodocrus ATCC21291 strain (4) Rhodococcus rhodocrus ATCC14347 strain (5) Rhodococcus rhodocrus ATCC14348 strain (6) Rhodococcus rhodocrus ATCC14349 strain (7) Rhodococcus rhodocrus ATCC14350 strain [9] The method according to [7] or [8], wherein the step is carried out under anaerobic conditions.

[10] The method according to any one of [7] to [9], wherein the hydrocarbon compound is a long-chain n-alkane of C12 to C44.

[11] The method according to

[10] , wherein the hydrocarbon compound is a long-chain n-alkane of C28 to C44.

[12] The method according to

[10] , wherein the hydrocarbon compound is octadecane.

[0009]

[13] A composition containing a microorganism for use in decomposing hydrocarbon compounds contained in soil and / or water, comprising: A composition, wherein the microorganism is one or more microorganisms belonging to any of the following species (1) to (3). (1) Gordonia neofelifaecis (2) Gordonia rubripertincta (3) Rhodococcus rhodochrous

[14] The microorganism is It is a microorganism of any of the following strains (1)-(7), or The composition of

[13] , which is a microorganism having 16S rRNA comprising a base sequence having 90% or more sequence identity with the strain and exhibiting the ability to decompose hydrocarbon compounds under anaerobic conditions. (1) Gordonia neoferrifaesis strain MCI526 (accession number: NITE P-03813) (2) Gordonia lubripeltinctula DSMZ43570 strain (3) Rhodococcus rhodocrus ATCC21291 strain (4) Rhodococcus rhodocrus ATCC14347 strain (5) Rhodococcus rhodocrus ATCC14348 strain (6) Rhodococcus rhodocrus ATCC14349 strain (7) Rhodococcus rhodocrus ATCC14350 strain

[15] The composition according to

[13] or

[14] , wherein the hydrocarbon compound is a long-chain n-alkane of C12 to C44.

[16] The composition according to

[15] , wherein the hydrocarbon compound is a long-chain n-alkane of C28 to C44.

[17] The composition of

[15] , wherein the hydrocarbon compound is octadecane.

[18] A composition containing a microorganism for use in decomposing hydrocarbon compounds contained in soil and / or water by the microorganism, comprising: The microorganism is Rhodococcus rhodocrustacean ATCC21291 strain, or A microorganism having 16S rRNA containing a base sequence having 90% or more sequence identity with the strain, and exhibiting an ability to decompose hydrocarbon compounds under anaerobic conditions. The hydrocarbon compound contains a long-chain n-alkane of C28 to C44. composition.

[0010]

[19] Gordonia neoferrifaesis MCI526 strain (accession number: NITE P-03813), or A microorganism belonging to Gordonia neoferriphaesis, having 16S rRNA containing a base sequence having 90% or more sequence identity with said strain, and exhibiting the ability to decompose hydrocarbon compounds under anaerobic conditions.

[20] The microorganism according to

[19] , wherein the hydrocarbon compound is a long-chain n-alkane having C12 to C44. [twenty one] The microorganism according to

[20] , wherein the hydrocarbon compound is a long-chain n-alkane of C28 to C44. [twenty two] The microorganism according to

[20] , wherein the hydrocarbon compound is octadecane. Effect of the Invention

[0011] The present disclosure provides a technology for decomposing hydrocarbon compounds contained in soil and / or water using microorganisms that exhibit the ability to decompose hydrocarbon compounds even under anaerobic conditions and pose few safety concerns. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] Preferred embodiments for carrying out the present disclosure will be described below. Note that the embodiments described below are examples of typical embodiments of the present disclosure, and the scope of the present disclosure should not be interpreted as being narrow.

[0013] [Hydrocarbon compound decomposition microorganisms] The hydrocarbon compound-decomposing microorganisms disclosed herein exhibit the ability to decompose hydrocarbon compounds under anaerobic conditions and can be used to decompose hydrocarbon compounds contained in soil and / or water (particularly soil and / or water with low oxygen concentrations).

[0014] In the present disclosure, the hydrocarbon compound is any compound consisting of carbon atoms (C) and hydrogen atoms (H), or these and other atoms, but is not limited thereto. It is preferably a long-chain n-alkane of C12 to C44, specifically dodecane, tridecane, tetradecane, pentadecane, hexadecane, heptadecane, octadecane, nonadecane, eicosane, heneicosane, docosane, tricosane, tetracosane, pentacosane, hexacosane, heptacosane, octacosane, nonacosane, hentriacontane, dotricontane, tritriacontane, tetratriacontane, pentatriacontane, hexatriacontane, heptatriacontane, octatriacontane, nonatriacontane, tetracontane, hentetracontane, dotetracontane, tritetracontane, and tetratetracontane. Of these, octadecane is known to be the main n-alkane contained in crude oil, and is therefore preferred from the viewpoint of purifying crude oil contamination. The hydrocarbon compound may be particularly a hydrocarbon compound having a carbon number of 28 to 44. The hydrocarbon compound-decomposing microorganism according to the present disclosure can exhibit high decomposition ability for such hydrocarbon compounds having a large number of carbon atoms per molecule.

[0015] In the present disclosure, anaerobic conditions refer to an environment in which the oxygen concentration is, for example, 1% or less, preferably 0.5% or less, more preferably 0.1% or less, and preferably 5% or more, more preferably 10% or more, and particularly preferably 15% or more of carbon dioxide gas concentration. The oxygen concentration can be measured by known methods, such as by galvanic cell sensors, polarographic sensors, and optical sensors.

[0016] In the present disclosure, the decomposition of a hydrocarbon compound is not particularly limited and may be decomposition by any route. As an example of an anaerobic alkane decomposition route, a route has been proposed in which one molecule of alkylsuccinic acid is synthesized from one molecule of n-alkane and one molecule of fumaric acid by alkylsuccinic acid synthase, two molecules of fatty acid are produced from the alkylsuccinic acid, and the fatty acid is decomposed by β-oxidation. The ability of microorganisms to decompose hydrocarbon compounds can be evaluated by contacting the hydrocarbon compounds with the microorganisms to cause a reaction, and then measuring the weight and molecular weight distribution of the hydrocarbon compounds before and after the decomposition reaction, or by analyzing their molecular structure. The decomposition ability of the hydrocarbon compound-decomposing microorganism according to the present disclosure shows an octadecane decomposition amount of 5% or more when the dry cell weight at the start of the culture is 1 to 4 mg / mL, the octadecane concentration is 1 mg / mL, and the microorganism is anaerobically cultured for 28 days at 30° C. The octadecane decomposition amount is preferably 6% or more, 7% or more, more preferably 8% or 9%, further preferably 10% or 11%, and particularly preferably 12% or more.

[0017] There are no particular limitations on the microorganism as long as it exhibits the ability to decompose hydrocarbon compounds under anaerobic conditions, but it is preferably one or more microorganisms belonging to any one of the following species (1) to (3): (1) Gordonia neofelifaecis (2) Gordonia rubripertincta (3) Rhodococcus rhodochrous

[0018] An example of a microorganism belonging to Gordonia neoferriphaesis is the Gordonia neoferriphaesis MCI526 strain (accession number: NITE P-03813). The MCI526 strain has been deposited domestically at the National Institute of Technology and Evaluation, Patent Microorganism Depositary (NPMD) (Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture), which is a depository institution under Article 27-2 and 3 of the Enforcement Regulations of the Patent Act and an international depository authority under the Budapest Treaty on the International Recognition of the Deposit of Microorganisms. The accession number of the above strain is NITE P-03813, and the date of receipt is January 26, 2023.

[0019] An example of a microorganism belonging to Gordonia lubripeltinctta is the Gordonia lubripeltinctta DSMZ43570 strain. The DSMZ43570 strain is available from the German Cell Bank (DSMZ-German Collection of Microorganisms and Cell Culture).

[0020] Examples of microorganisms belonging to the Rhodococcus rhodoclass include Rhodococcus rhodoclass ATCC21291 strain, ATCC14347 strain, ATCC14348 strain, ATCC14349 strain, and ATCC14350 strain. These strains are available from the American Type Culture Collection (ATCC).

[0021] The hydrocarbon compound-decomposing microorganism may be a closely related strain belonging to the same species as the above-mentioned strain, such as a microorganism belonging to the genus Gordonia or Rhodococcus, preferably Gordonia neoferrifaesis, Gordonia rubripeltincta, or Rhodococcus rhodoclass, and having a 16S rRNA containing a base sequence having 90% or more, preferably 93% or more, sequence identity with the above-mentioned strain. The nucleotide sequences of specific regions (about 1.5 kb) of 16S rRNA of the Gordonia neoferrifaecalis MCI526 strain and the Gordonia lubripeltincta DSMZ43570 strain are shown in SEQ ID NOs: 1 and 2. The nucleotide sequences of 16S rRNA of the Rhodococcus rhodocras ATCC21291 strain, ATCC14347 strain, ATCC14348 strain, ATCC14349 strain, and ATCC14350 strain are shown in SEQ ID NOs: 3-7. These closely related species are expected to be capable of decomposing hydrocarbon compounds under anaerobic conditions, similar to the above strains.

[0022] From the viewpoint of retaining hydrocarbon compound decomposition activity, the closely related strain has a 16S rRNA comprising a base sequence that is preferably 90% or more identical to the base sequence of a partial region of the 16S rRNA of the strain, more preferably 93% or more, 95% or more, 97% or more, and particularly preferably 99% or more identical to the base sequence of the entire region of the 16S rRNA of the strain, or has a 16S rRNA comprising a base sequence that is preferably 85% or more identical to the base sequence of the entire region of the 16S rRNA of the strain, more preferably 90% or more, 93% or more, 95% or more, 97% or more, and particularly preferably 99% or more identical to the base sequence of the entire region of the 16S rRNA of the strain. Hydrocarbon compound-degrading microorganisms can be obtained by selecting candidate microorganisms from among microorganisms belonging to the above-mentioned taxonomic genera or species based on sequence identity of the 16S rRNA base sequence, and evaluating the hydrocarbon compound decomposition ability of the candidate microorganisms. For the analysis and identification method of 16S rRNA, reference can be made to, for example, "Japanese Pharmacopoeia, 16th Edition; Rapid Identification Method of Microorganisms by Genetic Analysis."

[0023] Furthermore, the hydrocarbon compound-degrading microorganism may include genetically modified strains of the above strains. The genetic modification may be a modification for improving the expression level or activity of an enzyme that is actively involved in the decomposition of hydrocarbon compounds, or a modification for suppressing the expression level or activity of an enzyme that is involved in the inhibition of the decomposition of hydrocarbon compounds. The genetic modification may be performed according to conventionally known molecular biology techniques.

[0024] Normal culture conditions can be applied to the culture of hydrocarbon compound-degrading microorganisms. The culture conditions may be aerobic or anaerobic, but aerobic conditions are preferred when the purpose is to grow microorganisms. Conditions such as medium, temperature, and period can be set within the scope of conventional technology for each target microorganism.

[0025] The composition containing the cultured microorganism can be used in the method for decomposing hydrocarbon compounds or the method for purifying soil and / or water, which will be described below. The composition may contain the hydrocarbon compound-decomposing microorganism and a carrier that holds the microorganism in a state in which it can grow again. The carrier may be, for example, a culture medium or a cryopreservation solution.

[0026] [Method for decomposing hydrocarbon compounds, and method for purifying soil and / or water] The method for decomposing hydrocarbon compounds according to the present disclosure is a method for decomposing hydrocarbon compounds contained in soil and / or water using microorganisms, and includes a step of contacting soil and / or water containing hydrocarbon compounds with hydrocarbon compound-decomposing microorganisms. Furthermore, the method for purifying soil and / or water according to the present disclosure includes a step of contacting the soil and / or water with a hydrocarbon compound-decomposing microorganism and allowing the action of the microorganism to decompose the hydrocarbon compounds.

[0027] Contact between the hydrocarbon compound and the microorganism can be achieved, for example, by adding the microorganism or a composition containing the microorganism to soil or water. The microorganism or the composition containing the microorganism may be added together with a known microbial growth promoter or a hydrocarbon decomposition promoter. As the hydrocarbon compound-decomposing microorganism, any one or more of the above-mentioned microorganisms can be used in appropriate combination.

[0028] Since the hydrocarbon compound-decomposing microorganism according to the present disclosure grows under anaerobic conditions and exhibits the ability to decompose hydrocarbon compounds, the step of contacting the hydrocarbon compound with the microorganism can be carried out under anaerobic conditions, and the microorganism can be particularly suitably used for bioremediation of hydrocarbon compounds contained in soil and / or water with low acidity concentrations. That is, the method for decomposing hydrocarbon compounds and the method for purifying soil and / or water according to the present disclosure can decompose hydrocarbon compounds contained in soil and / or water deep underground where oxygen concentrations are low, without creating aerobic conditions by installing piping to supply air underground or periodically stirring the soil. The method for decomposing hydrocarbon compounds and the method for purifying soil and / or water according to the present disclosure can be suitably applied to soil deep below the ground surface where oxygen is difficult to be supplied, for example, soil below 5 m, or even below 10 m from the ground surface.

[0029] Furthermore, since the hydrocarbon compound-decomposing microorganisms disclosed herein are unlikely to cause serious illness in humans or animals, the hydrocarbon compound decomposition method and soil and / or water purification method disclosed herein can be suitably applied to bioremediation of outdoor contaminated soil in factories, gas stations, etc. EXAMPLES

[0030] [Example 1: Anaerobic octadecane vaporization supply test] Microorganisms properly held by the applicant and microorganisms properly obtained from microorganism holding institutions were inoculated onto OM2 synthetic medium agar plates. The compositions of the vitamin solution, trace element solution, and OM2 agar medium are shown in Table 1, Table 2, and Table 3, respectively. Filter paper was placed on the lid side of the petri dish and soaked in 2 mL of octadecane. An anaerobic environment (oxygen concentration below 0.1%, carbon dioxide concentration above 15%) was created using an oxygen absorber / carbon dioxide generator (Anelopac, Mitsubishi Gas Chemical Co.), and the microorganisms were cultured while supplying vaporized octadecane at 30°C.

[0031] [Table 1]

[0032] [Table 2]

[0033] [Table 3]

[0034] For strains that showed good growth potential, a specific region (approximately 1.5 kb) of 16S rDNA was amplified by PCR, and the species was identified by base sequence analysis. The colonies were suspended in 100 μL of 20 mM NaOH, heated at 98°C for 10 minutes, and 4 μL of 1 M Tris-HCl (pH 7.5) was added and centrifuged to obtain the supernatant. The PCR products were purified and sequenced, and the species was identified by a homology search of the sequence using the international base sequence database (Standard Nucleotide BLAST).

[0035] [Example 2: Anaerobic octadecane decomposition test] The test strains were aerobically cultured in LB medium at 30°C and 270 rpm for 3 to 4 days. The cells were harvested, washed with 1 mL of 0.9% NaCl, and suspended in 1 mL of 0.9% NaCl. 0.3 mL of the bacterial cell suspension containing 1 to 4 mg of dry bacterial cell weight was added to a test medium (OM3 medium) containing 1 g / L octadecane emulsified with 0.7 mL of Tween 80 to prepare 1 mL of test solution. The composition of the OM3 medium is shown in Table 4. The 15 mL glass tube containing the test solution was placed in an Anaeropack-containing square jar (manufactured by Mitsubishi Gas Chemical Co., Ltd.) under anaerobic conditions (oxygen concentration 0.1% or less, carbon dioxide concentration 15% or more) at 30°C to start the octadecane decomposition test. After 28 days, 9 mL of ethanol containing hexadecane (internal standard) was added to the test solution to extract the remaining octadecane, and the octadecane concentration was quantified by GC-FID.

[0036] [Table 4]

[0037] The results are shown in Table 5.

[0038] [Table 5]

[0039] No reduction in the amount of octadecane was observed in Gordonia terrae MCI484 and Pseudomonas stutzeri MCI2666, indicating that these strains did not exhibit the ability to decompose octadecane under anaerobic conditions. In addition, the reduction in octadecane content in Rhodococcus erythropolis MCI2259, Gordonia alkanivorans MCI2955, Rhodococcus erythropolis MCI2575 and Delftia acidovorans MCI4574 was less than 5%.

[0040] On the other hand, a decrease in the predominant octadecane amount of 5% or more was observed in Rhodococcus rhodochrous ATCC21291, Rhodococcus rhodochrous ATCC14347, Gordonia neoferifaecis MCI526, Rhodococcus rhodochrous ATCC14348, Gordonia rubripertincta DSMZ43570, Rhodococcus rhodochrous ATCC14349 and Rhodococcus rhodochrous ATCC14350, demonstrating that these strains grow under anaerobic conditions (Example 1) and exhibit excellent octadecane decomposition ability.

[0041] The sequence identity of the base sequences of the specific regions of 16S rDNA determined in Example 1 for these seven strains was calculated using the nucleic acid-to-nucleic acid homology search function of GenetyxRver.17 (Japan Server Co., Ltd.), and the results are shown in Table 6. The lowest sequence identity was 93% and the highest was 99%.

[0042] [Table 6]

[0043] [Example 3: Decomposition test of high carbon number hydrocarbon compounds other than octadecane] Rhodococcus rhodochrous ATCC21291 was spread on LBG agar medium (LB medium, 2% glucose, 1.5% agar) and cultured at 30°C for 3 days. 150 mL of LBG medium (LB medium + 2% glucose) was added to a 500 mL Erlenmeyer flask, and the strain grown on the plate was inoculated and cultured aerobically at 30°C and 130 rpm for 2 days. The bacteria were collected and washed with 300 mL of 0.9% sodium chloride aqueous solution, and then suspended in 300 mL of 0.9% sodium chloride aqueous solution. 300 mL of the bacterial suspension (containing 2.6 g of bacterial cells as dry weight), 250 mL of the medium shown in Table 7, 1 mL of the medium shown in Table 8, and 100 mL of oil-contaminated soil from the Fukaya oil field were added to a stirring device, and black horticultural soil (400-460 g) was added to make up to 1 L. 100 mL of oil-contaminated soil weighed approximately 170 g, and the measured value of total petroleum hydrocarbons (TPH) in the oil-contaminated soil was 15,000 mg / kg. The mixture was dispensed into small containers (140 mL) until they were full, and the containers were capped to prepare test samples. For comparison, a control sample was also prepared by replacing 300 mL of the bacterial suspension with 300 mL of water. After standing at room temperature for a period of time, the samples were subjected to TPH analysis.

[0044] [Table 7]

[0045] [Table 8]

[0046] The results are shown in Tables 9 and 10. Table 9 shows the measured values ​​of TPH for the test samples (containing Rhodococcus rhodochrous ATCC21291). Table 10 shows the measured values ​​of TPH for the control samples (not containing Rhodococcus rhodochrous ATCC21291).

[0047] [Table 9]

[0048] [Table 10]

[0049] After the second week, the test samples showed a decrease in TPH measurements. The decrease in TPH measurements was particularly notable for C28-C44 hydrocarbon compounds. This indicates that Rhodococcus rhodochrous ATCC21291 is particularly useful for decomposing hydrocarbon compounds with a large number of carbon atoms per molecule. [Sequence List Free Text]

[0050] SEQ ID NO: 1: Nucleotide sequence of 16S rRNA of Gordonia neoferrifaecis strain MCI526 SEQ ID NO: 2: Nucleotide sequence of 16S rRNA of Gordonia rubripeltinctta strain DSMZ43570 SEQ ID NO: 3: Nucleotide sequence of 16S rRNA of Rhodococcus rhodoclass ATCC21291 strain SEQ ID NO: 4: Nucleotide sequence of 16S rRNA of Rhodococcus rhodoclass ATCC 14347 strain SEQ ID NO: 5: Nucleotide sequence of 16S rRNA of Rhodococcus rhodoclass ATCC 14348 strain SEQ ID NO: 6: Nucleotide sequence of 16S rRNA of Rhodococcus rhodoclass ATCC 14349 strain SEQ ID NO: 7: Nucleotide sequence of 16S rRNA of Rhodococcus rhodoclass ATCC 14350 strain

Claims

1. A method for decomposing hydrocarbon compounds contained in soil and / or water by microorganisms, comprising the steps of: contacting the soil and / or the water containing the hydrocarbon compound with the microorganism, The method, wherein the microorganism is one or more microorganisms belonging to any of the following species (1) to (3): (1) Gordonia neofelifaecis (2) Gordonia rubripertincta (3) Rhodococcus rhodochrous

2. The microorganism is It is a microorganism of any one of the following strains (1) to (7), or The method according to claim 1, wherein the microorganism has 16S rRNA comprising a base sequence having 90% or more sequence identity with the strain and exhibits the ability to decompose hydrocarbon compounds under anaerobic conditions. (1) Gordonia neoferrifaesis MCI526 strain (accession number: NITE P-03813) (2) Gordonia lubripeltincta DSMZ43570 strain (3) Rhodococcus rhodocrus ATCC21291 strain (4) Rhodococcus rhodocrus ATCC14347 strain (5) Rhodococcus rhodocrus ATCC14348 strain (6) Rhodococcus rhodocrus ATCC14349 strain (7) Rhodococcus rhodocrus ATCC14350 strain

3. The method of claim 1 or 2, wherein the process is carried out under anaerobic conditions.

4. The method according to claim 1 or 2, wherein the hydrocarbon compound is a long-chain n-alkane of C12 to C44.

5. The method according to claim 4, wherein the hydrocarbon compound is a long chain n-alkane of C28 to C44.

6. 1. A method for remediating soil and / or water contaminated with hydrocarbon compounds, comprising: The method includes a step of contacting the soil and / or the water with microorganisms and decomposing the hydrocarbon compounds by the action of the microorganisms, The method, wherein the microorganism is one or more microorganisms belonging to any of the following species (1) to (3): (1) Gordonia neofelifaecis (2) Gordonia rubripertincta (3) Rhodococcus rhodochrous

7. A composition containing a microorganism for use in decomposing hydrocarbon compounds contained in soil and / or water by the microorganism, comprising: The composition, wherein the microorganism is one or more microorganisms belonging to any of the following species (1) to (3): (1) Gordonia neofelifaecis (2) Gordonia rubripertincta (3) Rhodococcus rhodochrous

8. Gordonia neoferrifaesis MCI526 strain (accession number: NITE P-03813), or A microorganism belonging to Gordonia neoferriphaesis, having 16S rRNA containing a base sequence having 90% or more sequence identity with said strain, and exhibiting the ability to decompose hydrocarbon compounds under anaerobic conditions.

Images