KR100641834B1 - A novel microorganism capable of decomposing petroleum and a method for decomposing petroleum therewith - Google Patents

Abstract

A novel microorganism capable of decomposing petroleum and a method for decomposing petroleum by using the same microorganism are provided to purify the environments polluted by petroleum without second pollution. The microorganism capable of decomposing petroleum Kribbia dieselivorans R33(KCTC 10801BP) is provided, wherein Kribbia dieselivorans R33(KCTC 10801BP) is isolated from the soil polluted by petroleum; and the petroleum is gasoline, kerosene, diesel oil, crude petroleum or lubricating oil. The soil polluted by petroleum is purified by treating it with Kribbia dieselivorans R33(KCTC 10801BP).

Description

A Novel Microorganism Capable of Decomposing Petroleum and a Method for Decomposing Petroleum Therewith}

1 is a Crivia diesel ribonance This diagram shows the molecular tree showing the evolutionary position of R33.

Figure 2 is a graph showing the diesel oil decomposition ability of microorganisms separated from the microbial community (MC-KY7) excellent in the decomposition activity of petroleum oil.

3 is the Crivia diesel ribonance This graph shows the decomposition of diesel oil over time of R33.

The present invention relates to a novel microorganism having a resolution of petroleum oil and a method of decomposing petroleum oil using the same, and more particularly, to a novel microorganism purely separated from a microbial community selected from oil contaminated soil. Crivia Diesel Ribolance in New Genus with Oil Resolution Kribbia dieselivorans R33 (R33), a method for decomposing petroleum oils using this novel microorganism and a method for the purification of oil pollution using the same.

Due to the rapid development of the domestic industry, the consumption of oil, which is used as an energy source and raw materials of various chemical products, increases exponentially, which increases the amount of oil storage facilities and transportation handling, causing concern about environmental damage.

Currently, more than 16,000 gas stations and 8,000 oil storage industry facilities are managed as oil pollution-causing facilities in Korea. In the case of foreign countries, it is known that in the case of gas stations, about 2% of facilities less than 5 years after completion and oil leaks occur in facilities that are 11 to 25 years old, and most oil storage facilities in Korea have been installed for more than 10 years. It is estimated that due to equipment aging and inadequate handling, many of these facilities leaked huge amounts of oil, contaminating the surrounding soil and groundwater.

In fact, as a result of measuring the soil pollution level of oil storage facilities, many facilities have received corrective orders exceeding the standards of concern in the Soil Environment Conservation Act, and representative examples of oil pollution include Busan Munhyeon district, near Anyang Indeokwon station, Incheon Munhaksan district, and Noksapyeong station in Seoul. US troops are included.

The soil contamination characteristics by oil are indirect, chronic and difficult to recover. In other words, when soil is contaminated, it leads to pollution of soil organisms including plants and groundwater, which returns to humans with great damage and has a long-term cumulative effect. In addition, soil pollution, like most environmental pollution, is difficult to improve, but requires much longer time and more economic investment than air or water quality.

The most common oils in gas stations and oil storage industry facilities are gasoline and diesel. Gasoline is easily volatilized into the air even if it is spilled into the soil because of its rapid volatilization, but light oil is not easily volatilized, and thus contaminates the soil while remaining long. There are petroleum-based diesel and tar-based diesel. Petroleum-based diesel is commonly referred to as diesel oil and diesel as a component that flows out from kerosene when distilling crude oil. Petroleum-based diesel oil is a mixture of various hydrocarbons, its boiling point is in the range of 200 to 350 ° C., its ignition point is at least 50 ° C. and its specific gravity is 0.830 to 0.880. The proportion of hydrocarbons is 75 to 80% of aliphatic hydrocarbons, 20 to 25% of aromatic hydrocarbons, 50% of aliphatic hydrocarbons are saturated alkanes, and the remainder is composed of alkyl side chain alkanes and cycloalkanes.

Various techniques for restoration of contaminated soil have been developed, including physicochemical and biological methods. Among these methods, biological methods can minimize secondary pollution compared with physicochemical methods. The core of the biological method is the decomposition of oil hydrocarbons by microorganisms, but the fact that petroleum hydrocarbons have a low bioavailability due to the hydrophobic nature and toxicity, so the degradation efficiency is very low.

Therefore, in recent years, in order to treat soil contaminated with petroleum oil, a method of activating indigenous microorganisms by injecting nutrients such as nitrogen and phosphorus into the contaminated place, or by injecting air, reapplying microorganisms separated from the contaminated place, and having excellent resolution Has been tried.

Therefore, the researchers secured an indigenous microbial community from the soils of oil-contaminated areas to restore petroleum oil-contaminated soils, and selected strain R33 with excellent oil resolution from the obtained microbial community. As a result of the isolation and identification of the selected strain R33, the present invention was completed by confirming that it is a new microorganism of a new genus not previously reported, and naming it Kribbia dieselivorans R33 .

An object of the present invention is to secure the indigenous microbial community (MC-KY7) from the soil in the oil contaminated area for the restoration of petroleum oil contaminated soil and separated from the secured microbial community identified petroleum oil resolution, especially diesel oil resolution New microorganism, Crivia Diesel Ribolance To provide R33.

Another object of the present invention is to stably and environmentally cleanse the soil contaminated with oil, and also the wastewater, seawater, or sewage, by using the oil resolution of the microorganisms to decompose pollutants.

It is another object of the present invention to provide an oil digester comprising said strain for use in the decomposition of oil contaminants.

Hereinafter, the present invention will be described in more detail.

In the present invention, a novel genus having a petroleum oil resolution, a new microorganism in the genus Kribbia , Crivia diesel riborance R33 (accession number: KCTC 10801BP) is provided.

In the present invention, as a result of securing the indigenous microbial community (MC-KY7) having oil degradation activity from the soil of petroleum oil contaminated area and searching for a novel microorganism expressing oil degradation ability from the obtained microbial community, It was selected as a strain showing good oil degradation activity. As a result of the isolation and identification of the finally selected strain, it was identified as a novel microorganism genus and species that has not been reported so far. Named R33.

In the present invention, "oil" means, but is not limited to, petroleum hydrocarbons such as gasoline, kerosene, diesel oil, heavy oil or lubricating oil.

Specifically, microorganisms having oil degradation activity according to the present invention were separated and identified by the following process: securing an indigenous microbial community with excellent oil degradation activity from tidal flats and soil samples near Gwangyang Works contaminated with petroleum oil. It was. In order to search for microorganisms having oil degradation activity from this indigenous microbial community, they were suspended in physiological saline, and then smeared and cultured in solid nutrient medium containing diesel oil. After 7 days, one strain having excellent oil degradation activity among the strains generated on the nutrient medium was selected and named R33.

After pure culture to identify the strains selected above, the form; Physiological and biochemical properties including utilization characteristics of a substrate that can be used as a carbon source and an energy source, and a property of generating an acid from carbohydrates; Chemical taxonomic characteristics such as the type of fatty acid; Sequence analysis of the 16S rRNA gene was performed.

According to the results of examining the morphology and physiological characteristics of strain R33, R33 is a gram positive aerobic bacteria. Growth conditions of the strain were tryticase soy agar (TSA) medium (Pancreatic Digest of casein 17.0 g, Enzymatic Digest of Soybean Meal 3 g, sodium chloride 5.0 g, K ₂ HPO ₄ 2.5 g / L, [difco 30.0 g / L], tryptic soybean agar) is preferably at 30 ℃, pH 7.0 to 8.0, physiological characteristics are oxidase negative, catalase positive, the ability to reduce nitrate to nitrite Confirmed. Also, alkaline phosphatase negative, esterase (C4) positive, lipase (C14) positive, leucine arylamidase positive, valine arylamidase negative, cysteine arylamidase negative, trypsin negative, alpha Chymotrypsin negative, acid phosphatase negative, naphthol-AS-BI-phosphohydrolase positive, alpha-galactosidase negative, beta-galactosidase positive, alpha-glucosidase positive, beta -Glucosidase positive, alpha-mannosidase negative. It decomposes esculin, Tween 80, but not urea, hypoxanthine, xanthine, tyrosine, and starch. Acids are produced from D-glucose, D-cellobiose, maltose, sucrose and D-trehalose, but D-sorbitol, Myo- inositol, D-xylose, D-ribose, D-fructose , D-mannitol, melibiose, L-arabinose, D-melezose, D-galactose, L-rhamnose, D-mannose, lactose, D-raffinose ) Did not produce acid.

In addition, as a result of observing the growth of the cells according to the sugar type of R33 and conducting sugar availability experiments as a carbon source and energy source, D-glucose, D-cellobiose, D-trehalose, sucrose, maltose, acetic acid, benzoic acid, pyruvic acid May be used as a carbon source and an energy source, but D-fructose, D-galactose, D-mannose, D-xylose, L-arabinose, citric acid, succinic acid, L-malate, and salicycin , Formic acid, and L-glutamate were not available as carbon and energy sources.

One of the chemical taxonomy of strain R33, the major fatty acids present in R33 are C _{16: 0} , iso-C _{16: 0} , 10-methyl-C _{16: 0} , C _{18: 1} ° C. 9c, 10-methyl- C _{18: 0} (TBSA). This is the main fatty acid of all community Nikko coarse genus (Genus Ornithinicoccus), intra-Castello is erasure in (Genus Intrasporangium), TB bakteo genus (Genus Terrabacter), in terra Rhodococcus (Genus Terracoccus) showing strain R33 homology with iso-C _{15: 0,} and the unsaturated fatty acid C _{17: 1} , It is also distinguished from Genus Knoellia , which has a small amount of C _{18: 1} (Groth et al ., Int. J. Syst. Bacteriol. , 49 , 1717, 1999; Int. J. Syst. Evol.Microbiol ., 52, 77, 2002; ... Maszenan et al, Int J. Syst Evol Microbiol 50, 593;.... Yoon et al, Int J. Syst Evol Microbiol, 50, 1821, 2000;.. Yoon et al ., Int. J. Syst. Evol. Microbiol. , 54 , 1975, 2004).

On the other hand, it is also different from Genus Janibacter with fatty acids of iso-C _{16: 0} , C _{17: 1} ω8c, C _{18: 1} ω9c, and C _{17: 0} (Yoon). et al ., Int. J. Syst. Evol. Microbiol. , 54 , 1975, 2004).

The nucleotide sequence of the 16S rRNA gene of strain R33 is shown in SEQ ID NO: 1.

Strain R33 is in Johnny bakteo showing molecular Systematic haksang, R33 and homology of the 16S rRNA base sequence (Genus Janibacter), greater Noel Ria in (Genus Knoellia), all community Nikko coarse genus (Genus Ornithinicoccus), TB bakteo in (Genus Terrabacter ), Genus Terracoccus , Genus Intrasporangium , compared to the standard strains of existing strains, showed a low homology of 95.9 to 93.8% and less than 97% of new species. Was identified as a new species not found so far based on international standards (Wayne et al. , Int. J. Syst. Bacteriol . 37 , 463, 1987). In the molecular system, Genus Janibacter , Genus Knoellia , Genus Ornithinicoccus , Genus Terrabacter , Genus Terracoccus , Intrasporane It has been identified as a new genus that does not exist because of its lack of flexible relation with Genus Intrasporangium . 1 is a molecular tree showing the evolutionary position of Crivia diesel riborance R33.

Therefore, in the present invention, the isolated strain was named Kribbia dieselivorans R33 and was deposited on May 16, 2005 with the Korea Biotechnology Research Institute Gene Bank (Accession Number: KCTC 10801BP).

The oil degradation activity of the new strain according to the present invention, Crivia diesel riborans R33, was measured as follows: Bushnell-Haas (BH) medium (0.2 g of MgSO ₄ , 0.02 g of CaCl ₂ , 1.0 g of KH ₂ PO ₄ ). , 1.0 g of K ₂ HPO _4, 1.0 g of KNO ₃ , 0.05 g / L of FeCl ₂ , [difco 3.27 g / L]), and 0.5 g (1%) of diesel oil were added as a substrate for decomposition at 30 ° C. After 10 days of incubation with reciprocating shaking at 120 rpm, the culture solution was extracted with the same amount of dichloromethane (DCM), and the remaining total petroleum hydrocarbon (TPH) was analyzed by gas chromatography (GC). The resolution is expressed as a percentage as a reference.

In the present invention, GC was used to analyze the content of TPH in a sample collected from an area contaminated with petroleum oil, and 3 ml of DCM was added to 1 g of a soil sample to perform GC, followed by mixing for 30 minutes. 1 g was added to remove moisture. The solvent layer was filtered through a 45 μm syringe filter and GC analysis was performed. GC analysis conditions were injection temperature 250 ℃, initial temperature of the column 40 ℃, final temperature 320 ℃, nitrogen was used as the carrier gas, operating time 38 minutes, the detector using a FID (flame ionization detector) oil decomposition Activity was measured.

The oil-degrading activity of Crivia Diesel Riborance R33 was very high (80%) as a result of measuring diesel oil-degrading activity with microorganisms purely isolated from the indigenous microbial community (MC-KY7) obtained from soils in oil-contaminated areas. Although it showed degrading activity, the control group and other isolated microorganisms which were not inoculated with microorganisms showed a relatively low degradation rate of 20 to 40%, indicating that the new strain Crivia diesel riborans R33 had excellent oil resolution.

Crivia diesel riborance R33 according to the present invention can be used as an oil decomposer in a biological method for purifying soil contamination by petroleum oil, in which case the crivia diesel riborance R33 according to the present invention is in liquid or powder form. It can be prepared as. In addition, in order to increase the treatment efficiency of the pollution purification by oil, the creevia digerylboranth R33 of the present invention may be manufactured and used in a fixed form by a conventional microorganism immobilization method.

The substrate of Crivia diesel riborance R33 according to the present invention is petroleum oil such as gasoline, kerosene, diesel oil, heavy oil or lubricating oil, preferably diesel oil.

By the following examples will be described in detail the present invention.

The following examples are intended to illustrate the invention, but are not intended to limit the scope of the invention.

Example 1 Isolation of Strains Having Oil Resolution

In order to isolate microorganisms having oil degradation activity, tidal flats and soil samples were collected near the Gwangyang Works contaminated with petroleum oil.

Bushnell-Haas (BH) medium (0.2 g of MgSO ₄ , 0.02 g of CaCl ₂ , 1.0 g of KH ₂ PO _4, 1.0 g of K ₂ HPO _4, 1.0 g of KNO ₃ , 0.05 g / L of FeCl ₂ , used as a separation medium, [difco 3.27g / L]) 50g of diesel oil 0.5g (1%) was added as a substrate for decomposition, 0.5g (1%) of soil sample was added and reciprocated shaking at 30 ° C at 120 rpm for 10 days Liver cultures.

The culture solution was extracted with the same amount of dichloromethane (DCM), and the remaining total petroleum hydrocarbon (TPH) was analyzed by gas chromatography (GC), and the primary oil resolution was expressed as a percentage based on the initial diesel oil concentration. The measurement secured an indigenous microbial community (MC-KY7) with over 80% oil resolution.

Suspended in 10 ml of saline solution (0.85% NaCl) to search for microorganisms with oil degradation activity from the obtained native microbial community (MC-KY7), and then R2A (0.5 g yeast extract, prothios peptone) with diesel oil added (Proteose Peptone No.3) 0.5 g, Casamino Acids 0.5 g, Dextrose 0.5 g, Soluble Starch 0.5 g, Sodium Pyruvate 0.3 g, K ₂ HPO ₄ 0.3 g, Magnesium Sulfate 0.05 g, Agar 15.0 g / L) Stained and cultured in solid nutrient medium. After 7 days, the strains generated on the nutrient medium were selected, and the remaining TPH of the selected strains was analyzed using gas chromatography. Based on the initial diesel oil concentration, the resolution was expressed as a percentage to confirm the diesel oil resolution, and one strain having excellent diesel oil degradation activity was selected and named as R33.

As a result of measuring the oil degradation activity of the strain R33 by measuring the oil degradation activity with microorganisms purely isolated from the indigenous microbial community (MC-KY7) selected from the soil of oil contaminated area, very high degradation of about 80% Control and other isolated microorganisms that showed activity but were not inoculated with microorganisms had relatively low degradation rates of 20-40%.

Example 2 Identification of Strains with Separated Oil Resolution

The strains isolated and selected in Example 1 were incubated in nutrient medium (TSA medium, manufactured by Difco) for 2 to 3 days at 30 ° C., and then examined for morphology, physiological and biochemical properties, chemical classification and molecular biological properties. It was.

(1) Morphology and Physiological and Biochemical Characteristics

Strain Morphological and physiological properties of R33 were determined using the method of Yoon et al., Int. J. Syst. Bacteriol. , 47 , 904, 1997 and the API ZYM kit (Biomerio). The production of acid from carbohydrates was done using Hugh & Leifson, Int. J. Syst. Bacteriol. , 48 , 907, 1998.

As a result of examining the morphology and growth conditions of strain R33, R33 is a gram positive aerobic bacterium and grew well under the temperature of 30 ° C. and pH 7.0-8.0. General physiological and biochemical characteristics and selected carbon source (energy source) utilization characteristics of the selected strain R33 are shown in Table 1 below.

 Physiological and Biochemical Characteristics and Selected Carbon Sources of Selected Strains  Characteristics Selected Strains (R33) Enzyme Activity (API ZYM)  Oxidase voice  Catalase positivity  Alkaline phosphatase voice  Esterase (C4) positivity  Lipase (C14) voice  Leucine arylamidase positivity  Valine arylamidase voice  Cysteine arylamidase voice  Trypsin voice  Alpha-chymotrypsin voice  Exid phosphatase voice  Naphthol-AS-BI-phosphohydrolase positivity  Alpha-galactosidase voice  Beta-galactosidase positivity  Alpha-glucosidase positivity  Beta-glucosidase positivity  Alpha-mannosidase voice  Nitrate reducing ability positivity Positive enzymatic activity; No negative enzyme activity Hydrolysis  Esculin +  Casein +  starch -  Twin 80 +  Element -  Hypersantin -  Santin -  Tyrosine +  + Degradable; -Cannot be disassembled Carbon source  D-glucose +  D-fructose -  D-galactose -  D-cellobiose +  D-Mannose -  D-trehalose -  D-Xylose -  L-Arabinose -  Suros +  Maltose +  Acetic acid +  Citric acid -  Succinic acid -  Benzoic acid +  L-apple acid -  Pyruvic acid +  Salinity -  L-glutamate - + Carbon source available; -No carbon source available

As shown in Table 1, strain R33 was oxidase negative, catalase positive, and had the ability to reduce nitrate to nitrite. It decomposes esculin, Tween 80, but not urea, hypoxanthine, xanthine, tyrosine, and starch. Acids are produced from D-glucose, D-cellobiose, maltose, sucrose, D-trehalose, but D-sorbitol, Myo- inositol, D-xylose, D-ribose , D-fructose, D-mannitol, melibiose, L-arabinose, D-melezose, D-galactose, L-rhamnose, D-mannose, No acid was produced from lactose, D-rapinose.

In addition, by examining the growth of the bacteria according to the sugar type of strain R33, as a carbon source and an energy source, sugar availability experiments were conducted. As a result, D-glucose, D-cellobiose, D-trehalose, sucrose, horse Toss, acetic acid, benzoic acid, pyruvic acid can be used as carbon and energy sources, but D-fructose, D-galactose, D-mannose, D-xylose, L-arabinose, citric acid, succinic acid, L-peracid (L- malate, salicycin, formic acid, and L-glutamate were not available as carbon and energy sources.

(2) Chemical Classification and Sequence Analysis of 16S rRNA Genes

The chemistry and the sequencing and analysis of 16S rRNA genes, including the fatty acid species, were performed by Yoon et al . (Yoon et al . Int. J. Syst. Bacteriol. , 47 , 933, 1997). The major fatty acids present in strain R33 are detailed in the form of C _{16: 0} , iso-C _{16: 0} , 10-methyl-C _{16: 0} , C _{18: 1} ω9c, 10-methyl-C _{18: 0} (TBSA) information are as shown in Table 2, which R33 and in all community Nikko coarse showing homology (genus Ornithinicoccus), intra-Castello is erasure in (genus Intrasporangium), TB bakteo genus (genus Terrabacter), in terra Rhodococcus (genus Terracoccus ) Is the major fatty acid of iso-C _{15: 0} and C _{17: 1} , It is also distinguished from Genus Knoellia , which has a small amount of C _{18: 1} (Groth et al ., Int. J. Syst. Bacteriol. , 49 , 1717, 1999; Int. J. Syst. Evol.Microbiol ., 52, 77, 2002; ... Maszenan et al, Int J. Syst Evol Microbiol 50, 593;.... Yoon et al, Int J. Syst Evol Microbiol, 50, 1821, 2000;.. Yoon et al ., Int. J. Syst. Evol. Microbiol. , 54 , 1975, 2004).

It also shows a clear difference from Genus Janibacter , which contains fatty acids of iso-C _{16: 0} , C _{17: 1} ω8c, C _{18: 1} ω9c, and C _{17: 0} , showing the potential as a new genus. Yoon et al ., Int. J. Syst. Evol. Microbiol. , 54 , 1975, 2004).

Fatty Acid Composition of Strain R33 Fatty acid types Furtherance (%) C _{14: 0}  2.10 C _{15: 0}  0.81 C _{16: 0} 11.30 C _{17: 0}  4.12 C _{18: 0} 11.01 iso-C _{14: 0}  0.60 iso-C _{15: 0}  0.80 iso-C _{16: 0} 15.63 iso-C _{17: 0}  0.65 iso-C _{18: 0}  2.41 C _{17: 1} ω8c  1.49 C _{18: 1} ω7c  1.09 C _{18: 1} ω9c 15.29 10-methyl C _{16: 0}  8.08 10-methyl C _{17: 0}  2.68 10-methyl C _{18: 0} 17.89 C _{16: 1} ω7c and / or iso-C _{15: 0} 2-OH  4.05

Strain R33 is in Johnny bakteo showing molecular Systematic haksang, R33 and homology of the 16S rRNA base sequence (Genus Janibacter), greater Noel Ria in (Genus Knoellia), all community Nikko coarse genus (Genus Ornithinicoccus), TB bakteo in (Genus Terrabacter ), Genus Terracoccus , Genus Intrasporangium showed a low homology of 95.9 to 93.8% compared to the standard strains of existing species.

Thus, up to 97% homology was identified as a new species that did not exist based on the international standard of new species (Wayne et al. , Int. J. Syst. Bacteriol . 37 , 463, 1987). Genus Janibacter , Genus Knoellia , Genus Ornithinicoccus , Genus Terrabacter , Genus Terracoccus , Intrasporangium (Genus Intrasporangium ) has a low flexibility relationship with the genus was identified as a new genus that does not exist (Fig. 1). The 16S rRNA gene nucleotide sequence of R33 was analyzed and shown in SEQ ID NO: 1, and was registered with GenBank, Accession No. DQ062659.

Based on the morphology, physiological and biochemical properties, chemical classification and 16S rRNA sequencing as described above, the isolated strain was identified as a new species of a new genus that has not been published previously. Kribbia dieselivorans R33) and was deposited with the Korea Biotechnology Research Institute Gene Bank on May 16, 2005 (accession number: KCTC 10801BP).

Example 3 Crivia Diesel Ribolance Oil resolution test of R33

(1) Comparison test of diesel oil resolution of Crivia Diesel Ribolance R33

Crivia Diesel Ribolance Separated and Identified in Examples 1 and 2 In order to investigate the oil-degrading activity of R33, 0.5 g (1%) of diesel oil was added to the separation medium according to Example 1 as a substrate for decomposition. After incubation for 10 days with reciprocating shaking at 30 ° C. at 120 rpm, the culture solution was extracted with the same amount of DCM, and the remaining TPH was analyzed using GC. The resolution was expressed as a percentage based on the initial diesel oil concentration.

GC was used to analyze the content of TPH in samples taken from areas contaminated with petroleum oil. To perform GC, add 3 ml of DCM to 1 g of soil sample, mix for 30 minutes, and add 1 g of sodium sulfate to remove moisture. The solvent layer was filtered through a 45 μm syringe filter and GC analysis was performed.

GC analysis conditions

Injection temperature: 250 ℃

Initial temperature of the column: 40 ℃

Final temperature: 320 ℃

Carrier Gas: Nitrogen

Driving time: 38 minutes

Detector: FID (flame ionization detector)

Diesel oil degradation activity was measured for the microorganisms (N54, N22, R33, R42, R44) purely separated from the obtained microbial community, and the results are shown in FIG. 2.

As shown in Figure 2, strain R33 showed lower degradation activity than the microbial community MC-KY7, but showed a very high degradation activity of about 80%, control and non-inoculated control microorganisms and other isolated microorganisms of about 20 to 40% Low degradation rate was shown.

(2) Diesel oil resolution over time of Crivia Diesel Riborance R33

Taking only the Crivia diesel riborance R33 having the best diesel oil resolution, the diesel oil decomposition pattern was investigated over time, and the results are shown in FIG. 3.

As shown in FIG. 3, when analyzed by the first order rate coefficient ( k ), the decomposition rate was very high as 0.234 (r ² = 0.953) .In strain R33, about 80% of diesel oil was decomposed at 6 days after inoculation. Since then no further decomposition has taken place. On the other hand, in the control group, about 30% of diesel oil naturally decreased due to volatilization after 10 days.

Crivia Diesel Ribolance from the results as described above R33 was confirmed that the oil resolution is excellent.

As described above, the present invention is a novel microorganism having a decomposition activity of petroleum oil, Crivia diesel riborance R33 ( Kribbia dieselivorans R33).

Crivia Diesel Ribolance of the invention R33 can effectively break down petroleum-based hydrocarbons and other oils, substances that are released into the natural environment during various industrial activities, causing environmental pollution and further harmful effects on humans and ecosystems.

Therefore, when applying Crivia diesel riborance R33 according to the present invention to various environments such as soils contaminated with petroleum hydrocarbons, unlike the conventional physicochemical soil purification method, the biodegradation process using microorganisms is stable and eco-friendly without secondary pollution. Can clean the environment.

Attach sequence list electronic file  

Claims (7)

Crivia Diesel Ribolance with Diesel Oil Resolution R33 (KCTC 10801BP). A method for decomposing petroleum oil components using the microorganism according to claim 1. 3. The method of claim 2 wherein the petroleum oil component is diesel oil. A petroleum oil decomposition agent comprising the microorganism according to claim 1. 5. The oily disintegrant of claim 4, wherein said petroleum oil disintegrant is used to treat oil contamination of soil. The oil decomposing agent according to claim 4 or 5, wherein the petroleum oil is diesel oil. The oil degradation agent of claim 4 in the form of a powder or liquid formulation.

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