CN101580808A - Rhodococcus ruber and application thereof in degradation of hydrocarbon compounds - Google Patents
Rhodococcus ruber and application thereof in degradation of hydrocarbon compounds Download PDFInfo
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- CN101580808A CN101580808A CNA2008100281226A CN200810028122A CN101580808A CN 101580808 A CN101580808 A CN 101580808A CN A2008100281226 A CNA2008100281226 A CN A2008100281226A CN 200810028122 A CN200810028122 A CN 200810028122A CN 101580808 A CN101580808 A CN 101580808A
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Abstract
The invention relates to a bacterial strain of Rhodococcus ruber P14 CGMCC NO.2343. The bacterial strain has the characteristic of floating up from oil matters; the bacterial strain can grow by taking the oil matters as unique carbon source and energy source and degrade the oil matters; and the bacterial strain can grow by taking polycyclic aromatic hydrocarbon as unique carbon source and energy source and degrade hydrocarbon compounds, such as phenanthrene, pyrene, benzopyrene, and the like. The bacterial strain can degrade the oil matters and hydrocarbon compounds, especially the polycyclic aromatic hydrocarbon can be applied in the biological treatment of oily waste water and the biological repair (biological remediation) of oil-contaminated soil.
Description
Technical field
The invention belongs to microorganism biological technology and Environmental Biotechnology field, specifically, relating to its deposit number of Rhodococcus ruber Rhodococcus ruber P14 is: CGMCC NO.2343, this bacterium can be floated in oily substance, the oily substance of degradable hydrocarbon-containifirst compound behind multiplication culture.
Background technology
Along with expanding economy, human demand to the energy also constantly enlarges, and each state has all accelerated the development and use to hydrocarbon resources, and from the desert to the ocean, from the depopulated zone to the densely populated area, increasing oil gas well appears at all parts of the world.The complex mixture that oil is made up of the thousands of kinds of different materials of chemical property mainly comprises stable hydrocarbon, compound fragrant hydrocarbon, bituminous matter, resene etc.The pollution of the exploitation of oil, smelting, use and transportation and omission accident, and the discharging of oily(waste)water, sewage irrigation, the volatilization of various petroleum products, incomplete combustion thing descend slowly and lightly etc. and to cause a series of petroleum pollution problems.
Oil and petroleum products mainly contain ocean, rivers and lakes, groundwater pollution to the pollution of water body.As forming oil film at the water surface, hindered the gaseous interchange between water body and the atmosphere, oils sticks on fish, algae and the planktonic organism, causes marine organisms death, and destroys seabird living environment, causes the dead and population quantity decline of seabird.Petroleum pollution also can make the fishery products quality descend, and causes financial loss.The rivers and lakes water pollution mainly is that waste water and the petroleum products of being refined the oil generation cause.In petroleum refining industry, there are a large amount of oily(waste)waters to discharge, because quantity discharged is big, often exceed the self-purification capacity of water body, easily form oil pollution.In addition, the principal pollutant greasy dirt that oil tanker washwater and boats and ships are produced when navigating by water in the waters also can pollute the waters.These pollutions are formed physics, chemical property or the coenosis of river, water body in lake and bed mud and are changed, thereby have reduced the use value of water body, even jeopardize people's health.
Bioremediation technology is to have occurred since the eighties in 20th century and the removing of development and the biotechnology of curbing environmental pollution, it mainly utilizes the ability of biospecific decomposition hazardous and noxious substances, remove the pollutent in contaminate environment such as the soil, reach the purpose of removing environmental pollution.In the budding stage of this technology, be mainly used in the improvement that environment PetroChina Company Limited. hydrocarbon pollutes, and achieve success.Practice result shows that bioremediation technology is feasible, effective and superior, and after this this technology is by the constantly improvement of broadened application other pollution types in environment.
Not only environmental protection but also oil removal method completely when though biological degradation is described as, in actually operating since bacterial classification can't effectively play a role at the pollution scene, but and action row is not strong.Major cause is as follows: at first be that oily substance is lighter than water mostly, float over the water layer surface impurely, rapidly diffusion and cause Polluted area to enlarge of the oily substance that leaks, this has brought very large influence for the microbiological deterioration oil pollutant.Secondly, in biodegradation process, biomass causes the biomass of useful effect to reduce, thereby causes degradation efficiency to reduce owing to diluted by water body, has also improved the cost of repairing environment simultaneously.
Rhodococcus is aerobic, the gram-positive microorganism of a class, because the multiple organic pollutant of this class bacterium energy metabolism, as arene, steroid, chlorination phenoplast, coal and oil or the like.Therefore rhodococcus is having broad application prospects aspect the environmental organism reparation.Rhodococcus has been applied to the processing of brown coal, high sulphur coal and oil now, reduces the viscosity of heavy oil and improves recovery utilization rate of oil or the like.In the oil pollutant, to the environmental hazard maximum be wherein hydrocarbon compound class material, present rarer bibliographical information rhodococcus is to hydrocarbon compound, the especially Degradation of high-molecular-weight hydrocarbons compounds.
Summary of the invention
The objective of the invention is at petroleum pollution provides a Rhodococcus ruber Rhodococcus sp. to the harm that environment causes, and this bacterium can degradation of hydrocarbon compounds, can be used for administering the bioremediation technology of petroleum pollution.This bacterium has the characteristic of floating in oily substance, can grow as sole carbon source with oily substance.
Rhodococcus ruber Rhodococcus sp. provided by the invention, be preserved in " China Committee for Culture Collection of Microorganisms's common micro-organisms " center "; its deposit number is: CGMCC NO.2343; the depositary institution address is: Datun Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica on January 16th, 2008.
Rhodococcus ruber P14 of the present invention cultivated 2 days on the 2216E flat board, and colony characteristics is: bacterium colony is circular, drying, opaque, rat, and neat in edge, color is orange red; Cell morphological characteristic children cell in age is shaft-like, and the thalline difference in length is bigger, shows polymorphism, random arrangement, and incubation time becomes sphere, Gram-positive after increasing; Physiological and biochemical property has the VP positive, do not produce gemma or spore, oxidase negative, the catalase feminine gender, reduction nitrate can utilize following material as carbon source: cyclodextrin, D-fructose, D-seminose, dextrin, D-Lip river ketose, ribose, a-D-glucose, D-pectinose, D-N.F,USP MANNITOL, polysorbate40, tween 80, L-lactic acid, acetic acid, propionic acid, a-ketone valeric acid, a-hydroxybutyric acid, 6-O-D Glucopyranose acyl, pyruvic acid, β beta-hydroxy-butanoic acid, mono succinate methyl ester, Pyruvic Acid Methyl ester.Flat board of the present invention and electromicroscopic photograph are respectively as Fig. 1, and be shown in Figure 2.
Adopt colony polymerase chain reaction (PCR) method, directly get single bacterium colony lysate and carry out PCR as template.The primer that utilizes pcr amplification 16SrDNA is a pair of universal primer, 27f:5 '-AGAGTTTGATCCTGGCTCAG-3 ', 1492R:5 '-GGTTACCTTGTTACGACT-3 '.The PCR reaction conditions is: 94 ℃ of 3min; 94 ℃ of 30s, 55 ℃ of 30s, 72 ℃ of 2 min, 30 circulations; 72 ℃ of 10min.T carrier (Shanghai SangonTA clones test kit) will be connected behind the pcr amplification product purifying, sequence on sequencing result and the Genbank is carried out Blast and is analyzed, sequence length is 1500bp, and accession number is EF634456 in Genbank, and the present invention and Rhodococcus ruber have very big homology; Its 16S rDNA sequence is:
1 agagtttgat?cctggctcag?gacgaacgct?ggcggcgtgc?ttaacacatg?caagtcgaac
61?gatgaagccc?agcttgctgg?gtggattagt?ggcgaacggg?tgagtaacac?gtgggtgatc
121?tgccctgcac?ttcgggataa?gcctgggaaa?ctgggtctaa?taccggatag?gacctcggga
181?tgcatgttcc?ggggtggaaa?ggttttccgg?tgcaggatgg?gcccgcggcc?tatcagcttg
241?ttggtggggt?aacggcccac?caaggcgacg?acgggtagcc?ggcctgagag?ggcgaccggc
301?cacactggga?ctgagacacg?gcccagactc?ctacgggagg?cagcagtggg?gaatattgca
361?caatgggcgc?aagcctgatg?cagcgacgcc?gcgtgaggga?tgacggcctt?cgggttgtaa
421?acctctttca?gtaccgacga?agcgcaagtg?acggtaggta?cagaagaagc?accggccaac
481?tacgtgccag?cagccgcggt?aatacgtagg?gtgcgagcgt?tgtccggaat?tactgggcgt
541?aaagagctcg?taggcggttt?gtcgcgtcgt?ctgtgaaaac?ccgcagctcg?actgcgggct
601?tgcaggcgat?acgggcagac?ttgagtactg?caggggagac?tggaattcct?ggtgtagcgg
661?tgaaatgcgc?agatatcagg?aggaacaccg?gtggcgaagg?cgggtctctg?ggcagtaact
721?gacgctgagg?agcgaaagcg?tgggtagcga?acaggattag?ataccctggt?agtccacgcc
781?gtaaacggtg?ggcgctaggt?gtgggtttcc?ttccacggga?tccgtgccgt?agctaacgca
841?ttaagcgccc?cgcctgggga?gtacggccgc?aaggctaaaa?ctcaaaggaa?ttgacggggg
901?cccgcacaag?cggcggagca?tgtggattaa?ttcgatgcaa?cgcgaagaac?cttacctggg
961?tttgacatac?accggaccgc?cccagagatg?gggtttccct?tgtggtcggt?gtacaggtgg
1021?tgcatggctg?tcgtcagctc?gtgtcgtgag?atgttgggtt?aagtcccgca?acgagcgcaa
1081?cccttgtcct?gtgttgccag?cacgtaatgg?tggggactcg?caggagactg?ccggggtcaa
1141?ctcggaggaa?ggtggggacg?acgtcaagtc?atcatgcccc?ttatgtccag?ggcttcacac
1201?atgctacaat?ggccggtaca?gagggctgcg?ataccgcgag?gtggagcgaa?tcccttaaag
1261?ccggtctcag?ttcggatcgg?ggtctgcaac?tcgaccccgt?gaagtcggag?tcgctagtaa
1321?tcgcagatca?gcaacgctgc?ggtgaatacg?ttcccgggcc?ttgtacacac?cgcccgtcac
1381?gtcatgaaag?tcggtaacac?ccgaagccgg?tggcctaacc?cctcgtggga?gggagccgtc
1441?gaaggtggga?tcggcgattg?ggacgaagtc?gtaacaaggt?aaccaagact?ggagatctg。
Sequence among this sequence and the Genbank is compared, choose similar bacterial strain, with Flip software building systematic evolution tree, constructed dendrogram as shown in Figure 4.
Rhodococcus ruber P14 of the present invention can be sole carbon source growth with the oily substance, the consisting of of its substratum: (NH
4)
2SO
4, 1.0g; Na
2HPO
4, 0.8g; KH
2PO
4, 0.2g; MgSO
47H
2O, 0.2g; CaCl
22H
2O, 0.1g; FeCl
36H
2O5mg; (NH
4)
6Mo
7O
244H
2O, 1mg; Distilled water 1000ml; Oils 40ml; PH7.0,121 ℃ of sterilization 30min.
Rhodococcus ruber P14 of the present invention can be sole carbon source growth with the polycyclic aromatic hydrocarbons, the consisting of of its substratum: (NH
4)
2SO
4, 1.0g; Na
2HPO
4, 0.8g; KH
2PO
4, 0.2g; MgSO
47H
2O, 0.2g; CaCl
22H
2O, 0.1g; FeCl
36H
2O5mg; (NH
4)
6Mo
7O
244H
2O, 1mg; Distilled water 1000ml; Polycyclic aromatic hydrocarbons 50mg; PH7.0,121 ℃ of sterilization 30min.
The present invention also provides Rhodococcus ruber P14 to use in degradation of hydrocarbon compounds, the particularly application in the polycyclic aromatic hydrocarbons in degradation of hydrocarbon, therefore bacterial strain of the present invention can be applied in the biological treatment of oily(waste)water and/or the biological restoration of oil-polluted soils owing to energy degradation of hydrocarbon compounds spy.
Bacterial strain of the present invention can be at nutritional medium, as cultivating in 2216E, LB, the common beef broth, also can in the basic medium that with oily substance such as crude oil, diesel oil, paraffin is sole carbon source, grow, can also in the basic medium that with polycyclic aromatic hydrocarbons is sole carbon source, grow as phenanthrene, pyrene, benzopyrene, bacterial strain all can carry out aerobic growth between 25 ℃-35 ℃, optimum growth temperature is 30 ℃.
Bacterial strain of the present invention has the characteristic that oily substance floats.Cultivating bacterial strain P14 to OD in the 2216E substratum is 0.8, collect thalline, contain in the test tube of minimal medium that different oils are carbon source with being inoculated in after the liquid minimal medium washing three times, cultivate after 10 days, bacterial strain P14 can float to oil reservoir, also breed rapidly, big gun shape protuberance appears in the critical surface at inorganic salt nutrient solution and upper strata oily substance, and the orange (see figure 5) appears in the upper strata oily substance simultaneously; Water oil interface thalline quantity is maximum, the minimum (see figure 6) of thalline quantity of bottom.In addition, after containing the inorganic salt nutrient solution inoculation P14 bacterial strain 12h of 4% crude oil, the crude oil agglutination phenomenon appears; And control group (4% crude oil+inorganic salt nutrient solution, not inoculating strain) does not then change (see figure 7).
Bacterial strain of the present invention has the characteristic of degradation of oils material.Cultivating bacterial strain P14 to OD in the 2216E substratum is 0.8, collect thalline, contain in the triangular flask of minimal medium that different oils are carbon source (prescription of substratum is with embodiment 4) with being seeded to after the liquid minimal medium washing three times, 30 ℃, shaking table cultivation (150rpm) were cultivated respectively 15 days, 30 days.Contrast nonvaccinated parallel sample, with the petroleum ether extraction nutrient solution, weighting method is measured the content of residue oily substance.15 days degradation rates to diesel oil, paraffin of strain culturing are respectively: 17.8% and 25%.30 days degradation rates to diesel oil, paraffin of bacterial strain are respectively 29.5% and 22.5%.This bacterium to the degradation rate figure of diesel oil, paraffin respectively shown in Fig. 8,4-2.
Bacterial strain of the present invention has the ability of degradation of hydrocarbon compounds.Cultivating bacterial strain P14 to OD in the 2216E substratum is 0.8, collect thalline, contain in the minimal medium triangular flask that different polycyclic aromatic hydrocarbonss are carbon source with being seeded to after the liquid minimal medium washing three times, 30 ℃ of shaking tables are cultivated (150rpm) and were cultivated 30 days, by the degradation rate of HPLC analysis polycyclic aromatic hydrocarbons, the result as shown in Figure 5.Figure 10 result shows, bacterial strain P14 is through 30 days cultivation, can reach 43% to the degradation rate of 50mg/L phenanthrene; And to pyrene, the benzopyrene of 50mg/L, through 30 days cultivation, degradation rate can reach 34%, 30% (Figure 11,5-3) respectively.
Description of drawings
Fig. 1 is the dull and stereotyped growing state figure of bacterial strain Rhodococcus ruber P14 CGMCC NO.2343; And electromicroscopic photograph.
Fig. 2 is a bacterial strain P14 electromicroscopic photograph of the present invention.
Fig. 3 is the 16S rDNA sequence chart of bacterial strain P14 of the present invention.
Fig. 4 is the phyletic evolution tree graph of bacterial strain P14 of the present invention.
Fig. 5 floats in the oily substance pictorial diagram for bacterial strain Rhodococcus ruber P14 CGMCC NO.2343.A wherein: inorganic salt/paraffin/P14, B: inorganic salt/diesel oil/P14, C: inorganic salt/vegetables oil, D: inorganic salt/vegetables oil/P14, E: inorganic salt/whiteruss, F: inorganic salt/whiteruss/streptococcus aureus, G: inorganic salt/whiteruss/P14.
Fig. 6 is bacterial strain P14 distribution situation in the inorganic salt nutrient solution that contains 4% oil (diesel oil or whiteruss), and right figure be the situation that adopts whiteruss cultivation bacterial strain P14, and the degree of depth that begins to mark liquid level above the test tube downwards is 1,2,5,10cm.
Fig. 7 makes crude oil aggegation pictorial diagram for bacterial strain P14 of the present invention, and wherein A, D, E inoculation of medium bacterial strain P14 cultivate 12h; B, C be inoculating strain not.
Fig. 8 for bacterial strain P14 of the present invention to the degraded of diesel oil (4%), through this bacterium after 15 days and the increment cultivation in 30 days to diesel degradation rate.
Fig. 9 bacterial strain of the present invention is to the degraded of paraffin (4%), through the degradation rate of this bacterium after the cultivation of rising in value in 15 days and 30 days to paraffin.
Figure 10 for bacterial strain P14 of the present invention through 30 days cultivation, to the degradation rate of the phenanthrene of 50mg/L.
Figure 11 for bacterial strain P14 of the present invention through 30 days cultivation, to the degradation rate of the pyrene of 50mg/L.
Figure 12 for bacterial strain P14 of the present invention through 30 days cultivation, to the degradation rate of the benzopyrene of 50mg/L.
Concrete embodiment
In order to understand the present invention better, illustrate better by following embodiment, but be not limitation of the invention.
Embodiment 1: the separation screening of bacterial strain Rhodococcus ruber P14 CGMCC NO.2343 provided by the invention
Sample comes from the Xiamen sea area surface deposit, takes by weighing the 5g bottom silt, adds 50ml sterilized water vibration 2h, leaves standstill that to draw the limpid liquid in upper strata behind the 30min be that inorganic salt add the (composition of substratum: (NH in the polycyclic aromatic hydrocarbons substratum to enrichment medium
4)
2SO
4, 1.0g; Na
2HPO
4, 0.8g; KH
2PO
4, 0.2g; MgSO
47H
2O, 0.2g; CaCl
22H
2O, 0.1g; FeCl
36H
2O5mg; (NH
4)
6Mo
7O
244H
2O, 1mg; Distilled water 1000ml; Polycyclic aromatic hydrocarbons 50mg; PH7.0,121 ℃ of sterilization 30min), cultivate 15d after, switching 1ml cultivates a week to containing 50mg/l polycyclic aromatic hydrocarbons inorganic salt nutrient solution, is forwarded to the fresh 50mg/l polycyclic aromatic hydrocarbons inorganic salt nutrient solution that contains.Behind the triplicate, line separates (consisting of of substratum: yeast extract 1g on the 2216E substratum; Peptone 5g; High ferric phosphate 0.01g; Chen Haishui 1000ml; PH7.6-7.8,121 ℃ of sterilization 30min) after picking list bacterium colony line separates three times, with degradation bacteria strains with the freezing preservation of 20% glycerine.
The pure degradation bacteria strains that obtains is inoculated in the 2216E liquid nutrient medium and is cultured to OD=0.8, centrifugal collection thalline, the influence that the minimal medium washing is removed remaining 2216E carbon source to degradation experiment three times, drawing equivalent bacterium liquid adds in the polycyclic aromatic hydrocarbons substratum in inorganic salt, cultivate after 30 days for 30 ℃, with the dichloromethane extraction nutrient solution, the HPLC method detects polycyclic aromatic hydrocarbons residue content.Separate obtaining a strain, comprise that phenanthrene, pyrene, benzopyrene all have the bacterial strain than high degradation rate, i.e. Rhodococcus rubber P14 CGMCC NO.2343 polycyclic aromatic hydrocarbons.
Embodiment 2: the pcr amplification and the sequencing of the 16S rDNA gene of bacterial strain Rhodococcus ruber P14 CGMCC NO.2343 provided by the invention
Adopt colony polymerase chain reaction (PCR) method, directly get single bacterium colony lysate and carry out PCR as template.The PCR primer of amplification 16S rDNA is a pair of universal primer, 27f:5 '-AGAGTTTGATCCTGGCTCAG-3 ', 1492R:5 '-GGTTACCTTGTTACGACT-3 '.The PCR reaction conditions is: 94 ℃ of 3min; 94 ℃ of 30s, 55 ℃ of 30s, 72 ℃ of 2 min, 30 circulations; 72 ℃ of 10min.Will be behind the pcr amplification product purifying connect T carrier (Shanghai SangonTA clone test kit), serve extra large Ying Jun Bioisystech Co., Ltd and check order.Sequence on sequencing result and the Genbank is carried out Blast and is analyzed.Sequence length is about 1500bp, and accession number is EF634456 in Genbank, and bacterial strain of the present invention and Rhodococcus ruber have very big homology.
Embodiment 3: bacterial strain Rhodococcus ruber P14 CGMCC NO.2343 provided by the invention floats the characteristic in oily substance
Cultivating Rhodococcus ruber P14 CGMCC NO.2343 to OD in the 2216E substratum is 0.8, collect thalline, (a collection of nutrient solution places long 12cm in the 50ml inorganic salt nutrient solution that contains 4% diesel oil of sterilizing or paraffin with being seeded to after the MSM washing three times, in the test tube of diameter 4.5cm, another batch places the 150ml triangular flask), 30 ℃, 150rpm were cultivated 10 days.Contrast nonvaccinated parallel sample, observe the variation of the oily substance that floats on the substratum top layer, for the test tube culture sample, drawing each layer nutrient solution 10ul gradient dilution after the standing over night is 10
-3, 10
-6, 10
-9, get each diluent coating of 50 μ l and be incubated at the 2216E flat board.Count colony counts behind 30 ℃ of cultivation 48h.
Embodiment 4: bacterial strain Rhodococcus ruber P14 CGMCC NO.2343 provided by the invention is to the degraded of oily substance
Cultivating Rhodococcus ruber P14 CGMCC NO.2343 to OD in the 2216E substratum is 0.8, collect thalline, contain in the triangular flask of minimal medium that different oils are carbon source (prescription of substratum is with embodiment 4) with being seeded to after the liquid minimal medium washing three times, 30 ℃, shaking table are cultivated (150rpm) and were cultivated 10 days.Contrast nonvaccinated parallel sample, with the petroleum ether extraction nutrient solution, weighting method is measured the content of residue oily substance; The result as shown in Figure 4.Fig. 8 shows, bacterial strain 15 days, 30 days is respectively 17.8%, 22.5% to diesel degradation rate; Fig. 9 shows that 15 days, 30 days degradation rates to paraffin of bacterial strain are respectively 25%, 29.5%;
Embodiment 5: bacterial strain Rhodococcus ruber P14 CGMCC NO.2343 provided by the invention is to the Degradation of polycyclic aromatic hydrocarbons
Get the fresh slant strains of bacterial strain of the present invention, be inoculated in the 2216E liquid nutrient medium with transfering loop and be cultured to OD0.8, centrifugal collection thalline, the influence that the minimal medium washing is removed remaining 2216E carbon source to degradation experiment three times, drawing equivalent bacterium liquid contains in the minimal medium triangular flask that the distinct fragrance hydrocarbon is a carbon source in 20ml is housed, 30 ℃ of shaking tables are cultivated (150rpm) and were cultivated 30 days, measure the increment (LgCFU/ml) of bacterium and the degraded (HPLC) of polycyclic aromatic hydrocarbons; The result as shown in Figure 5.Figure 10 shows that to the phenanthrene of 50mg/L, through 30 days cultivation, degradation rate can reach 43%; Figure 11 shows that to the pyrene of 50mg/L, through 30 days cultivation, degradation rate can reach 34%; Figure 12 shows that to the benzopyrene of 50mg/L, through 30 days cultivation, degradation rate can reach 30%.
Claims (8)
1. a strain has the Rhodococcus ruber Rhodococcus ruber P14 of degradation of hydrocarbon compounds, and its deposit number is: CGMCC NO.2343.
2. Rhodococcus ruber P14 according to claim 1 is characterized in that the nucleotide sequence of its 16S rDNA is as follows:
1?agagtttgat?cctggctcag?gacgaacgct?ggcggcgtgc?ttaacacatg?caagtcgaac
61?gatgaagccc?agcttgctgg?gtggattagt?ggcgaacggg?tgagtaacac?gtgggtgatc
121?tgccctgcac?ttcgggataa?gcctgggaaa?ctgggtctaa?taccggatag?gacctcggga
181?tgcatgttcc?ggggtggaaa?ggttttccgg?tgcaggatgg?gcccgcggcc?tatcagcttg
241?ttggtggggt?aacggcccac?caaggcgacg?acgggtagcc?ggcctgagag?ggcgaccggc
301?cacactggga?ctgagacacg?gcccagactc?ctacgggagg?cagcagtggg?gaatattgca
361?caatgggcgc?aagcctgatg?cagcgacgcc?gcgtgaggga?tgacggcctt?cgggttgtaa
421?acctctttca?gtaccgacga?agcgcaagtg?acggtaggta?cagaagaagc?accggccaac
481?tacgtgccag?cagccgcggt?aatacgtagg?gtgcgagcgt?tgtccggaat?tactgggcgt
541?aaagagctcg?taggcggttt?gtcgcgtcgt?ctgtgaaaac?ccgcagctcg?actgcgggct
601?tgcaggcgat?acgggcagac?ttgagtactg?caggggagac?tggaattcct?ggtgtagcgg
661?tgaaatgcgc?agatatcagg?aggaacaccg?gtggcgaagg?cgggtctctg?ggcagtaact
721?gacgctgagg?agcgaaagcg?tgggtagcga?acaggattag?ataccctggt?agtccacgcc
781?gtaaacggtg?ggcgctaggt?gtgggtttcc?ttccacggga?tccgtgccgt?agctaacgca
841?ttaagcgccc?cgcctgggga?gtacggccgc?aaggctaaaa?ctcaaaggaa?ttgacggggg
901?cccgcacaag?cggcggagca?tgtggattaa?ttcgatgcaa?cgcgaagaac?cttacctggg
961?tttgacatac?accggaccgc?cccagagatg?gggtttccct?tgtggtcggt?gtacaggtgg
1021?tgcatggctg?tcgtcagctc?gtgtcgtgag?atgttgggtt?aagtcccgca?acgagcgcaa
1081?cccttgtcct?gtgttgccag?cacgtaatgg?tggggactcg?caggagactg?ccggggtcaa
1141?ctcggaggaa?ggtggggacg?acgtcaagtc?atcatgcccc?ttatgtccag?ggcttcacac
1201?atgctacaat?ggccggtaca?gagggctgcg?ataccgcgag?gtggagcgaa?tcccttaaag
1261?ccggtctcag?ttcggatcgg?ggtctgcaac?tcgaccccgt?gaagtcggag?tcgctagtaa
1321?tcgcagatca?gcaacgctgc?ggtgaatacg?ttcccgggcc?ttgtacacac?cgcccgtcac
1381?gtcatgaaag?tcggtaacac?ccgaagccgg?tggcctaacc?cctcgtggga?gggagccgtc
1441?gaaggtggga?tcggcgattg?ggacgaagtc?gtaacaaggt?aaccaagact?ggagatctg。
3. Rhodococcus ruber P14 according to claim 1 is characterized in that: cultivated 2 days on the 2216E flat board, colony characteristics is: circular, dry, opaque, and rat, neat in edge, color is orange red; Cell morphological characteristic children cell in age is shaft-like, and the thalline difference in length is bigger, shows polymorphism, random arrangement, and incubation time becomes sphere, Gram-positive after increasing; Physiological and biochemical property has the VP positive, does not produce gemma or spore, oxidase negative, catalase feminine gender, reduction nitrate.
4. Rhodococcus ruber P14 according to claim 3: it is characterized in that: this bacterium can be sole carbon source growth with the oily substance, the consisting of of its substratum: (NH
4)
2SO
4, 1.0g; Na
2HPO
4, 0.8g; KH
2PO
4, 0.2g; MgSO
47H
2O, 0.2g; CaCl
22H
2O, 0.1g; FeCl
36H
2O5mg; (NH
4)
6Mo
7O
244H
2O, 1mg; Distilled water 1000ml; Oils 40ml; 7.0,121 ℃ of sterilizations of pH 30min.
5. Rhodococcus ruber P14 according to claim 1 is characterized in that this bacterium can be sole carbon source growth with the polycyclic aromatic hydrocarbons, the consisting of of its substratum: (NH
4)
2SO
4, 1.0g; Na
2HPO
4, 0.8g; KH
2PO
4, 0.2g; MgSO
47H
2O, 0.2g; CaCl
22H
2O, 0.1g; FeCl
36H
2O5mg; (NH
4)
6Mo
7O
244H
2O, 1mg; Distilled water 1000ml; Polycyclic aromatic hydrocarbons 50mg; 7.0,121 ℃ of sterilizations of pH 30min.
6. the described Rhodococcus ruber P14 of claim 1 uses in degradation of hydrocarbon compounds.
7. the application of the polycyclic aromatic hydrocarbons of the described Rhodococcus ruber P14 of claim 6 in degradation of hydrocarbon compounds.
8. the application of the described Rhodococcus ruber P14 of claim 1 in the biological restoration of the biological treatment of oily(waste)water and/or oil-polluted soils.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008100281226A CN101580808B (en) | 2008-05-15 | 2008-05-15 | Rhodococcus ruber and application thereof in degradation of hydrocarbon compounds |
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