CN107893037A - Oil and phenanthrene degradation bacteria strain P51 suitable for subsea cryogenic environment - Google Patents

Oil and phenanthrene degradation bacteria strain P51 suitable for subsea cryogenic environment Download PDF

Info

Publication number
CN107893037A
CN107893037A CN201711067118.6A CN201711067118A CN107893037A CN 107893037 A CN107893037 A CN 107893037A CN 201711067118 A CN201711067118 A CN 201711067118A CN 107893037 A CN107893037 A CN 107893037A
Authority
CN
China
Prior art keywords
oil
bacterial strain
degradation
strain
bacterial
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201711067118.6A
Other languages
Chinese (zh)
Other versions
CN107893037B (en
Inventor
刘秋
肖圣月
李佳怡
于基成
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Dalian Minzu University
Original Assignee
Dalian Minzu University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Dalian Minzu University filed Critical Dalian Minzu University
Priority to CN201711067118.6A priority Critical patent/CN107893037B/en
Priority to CN201711358029.7A priority patent/CN108046441B/en
Publication of CN107893037A publication Critical patent/CN107893037A/en
Application granted granted Critical
Publication of CN107893037B publication Critical patent/CN107893037B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/02—Separating microorganisms from their culture media
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00—Nature of the contaminant
    • C02F2101/30—Organic compounds
    • C02F2101/32—Hydrocarbons, e.g. oil
    • C02F2101/327—Polyaromatic Hydrocarbons [PAH's]

Abstract

The invention belongs to microbial technology field, and in particular to oil and phenanthrene degradation bacteria strain P51 suitable for subsea cryogenic environment.The bacterial strain deposit number is CGMCCNo.14411, its 16SrRNA sequence such as SEQ ID No：Shown in 1.The bacterial strain P51 that the present invention screens has the function of efficient degradation oil, bacterial strain P51 is while degraded oil, also there is higher degradability to polycyclic aromatic hydrocarbon China and Philippines, this characteristic solves caused marine pollution matter after oil pollution, marine environment is recovered completely, and laid the foundation for exploitation microorganism remediation oil pollution product.

Description

Oil and phenanthrene degradation bacteria strain P51 suitable for subsea cryogenic environment
Technical field
The invention belongs to microbial technology field, and in particular to one plant of bacterial strain P51 and profit with oil degradation function Marine Environmental Governance is carried out with the bacterial strain.
Background technology
With the development of the social economy, requirement of the mankind to oil constantly expands.Oil product such as gasoline, diesel oil, lubrication Oil, paraffin, pitch etc. gradually walk close to the life of the mankind, and all kinds of contamination accidents of the thing followed are also of common occurrence.2007 11 Month, the Russian oil tanker " volga oil 139 " for loading 4700 tons of heavy oil meets with blast in Kerch straits, sinking of disintegrating, More than 3000 tons of heavy oil spill, accident sea area is caused seriously to be polluted；On April 20th, 2010, semisubmersible drilling platform " deep water Horizon Line " explodes, and sinks to the Gulf of Mexico two days later, has about 10,000 tons of crude oil leakages daily to the Gulf of Mexico, causes extremely serious Oil pollution；On July 17th, 2010, the oil pipeline generation of the neighbouring petrochina of Dalian New Port is on fire from explosion, although Puted out a fire to save life and property after 15 hours, but this blast but result in substantial oil and be bled into sea；The CNOOC Bohai Sea during in June, 2011 Oil accident occurs for the oil field of gulf one, oil leak periods of months, caused by endanger and be unable to estimate.In mid or late November, 2013, positioned at green grass or young crops The yellow petroleum pipeline in sinopec east of island economic development zone occurs to rupture and explode, and this event not only causes dirt to local marine site Contaminate the security of the lives and property more to local resident and ecological environment causes huge loss.The explosion of offshore oil well, oil Endangered caused by the rupture of pipeline, marine environment, the health of the mankind and sustainable development are generated directly or indirectly Influence.According to statistics, the whole world there are about in 600~10,000,000 tons of oil and products thereof inflow water body every year, and wherein have absolutely It is most of to flow into ocean.
Oil pollution getting worse, the concern of various circles of society is triggered, the governing problem of oil pollution, has also become current The task that scholar primarily solves.Currently used method has physical treatment, chemical treatment and biological treatment.Physical treatment is to pass through The oil of ocean surface such as is diluted, assembled, migrating at the processing by physics mode, but can not thoroughly remove in seawater and dissolve Oil；Chemical treatment is that certain chemical reagent is added into seawater, reaches degraded purpose with this, but such a mode can draw Play secondary pollution；Biological treatment is then to utilize bioremediation technology, is entered the petroleum pollution in ocean by microbial metabolism Row degraded, has good in economic efficiency, treatment effeciency high and the small advantage of environmental pollution, can fundamentally solve asking for oil pollution Topic.For bioremediation technology for other processing methods, expense is relatively low, and economic benefit and ring benefit are all good, will turn into one Kind solves the effective ways of complex environment pollution problem.
Current study show that the microorganism for being capable of degraded oil has 79 category, totally 200 multiple-microorganism, they belong to respectively In bacterium, actinomyces, mould, saccharomycete and algae.Wherein mainly there is most common oil degradation bacteria in soil：Microbacterium (Microbacterium), Joint core bacterium category (Arthrobacter), Micrococcus (Micrococcus), streptomyces (Streptomyces) etc., most important degradation bacteria has in ocean：Achromobacter (Achromobacter), acinetobacter calcoaceticus Belong to (Acinetobacter), alcaligenes (Alcaligenes) etc..
The more microbial treatment environment oil pollution of document report at present is concentrated mainly on land oil pollution.It is such as former Know gorgeous[1]3 plants of bacterial strains using crude oil as sole carbon source are isolated from petroleum-polluted seawater, identified 3 plants of bacterial strains are respectively Belong to γ-deformation Gammaproteobacteria and Actinomycetes, wherein γ-deformation Gammaproteobacteria degradation rate highest, can reach 30.52%.By these three bacterium Respectively during combination of two culture, its combination strain is above independent bacterial strain to the degradation rate of crude oil, and not equal to each corresponding single bacterium Degradation rate sum.It can be seen that combination strain than single bacterium can more effective decomposing petroleum hydrocarbon, due to competition, short of money between each bacterial strain The effect of the various factors such as anti-, promotion, form the degradation rate of Mixed Microbes.Guo Qianyu etc.[2]2 plants of advantages are obtained by screening to drop Oily bacterium, through Preliminary Identification, one plant is bacillus, and another strain is acinetobacter.And by changing temperature, pH, inoculum concentration etc. Different condition, finally draw the optimal degradation condition of two plants of bacterium.Wang Jianan etc.[3]From Tianjin Dagang Oilfield oil-polluted soils and 6 plants of Crude Oil-degrading Bacterias are isolated in the offshore drilling platform washing oil sewage of the Bohai Sea, it is identified to be belonging respectively to Bacillus, false list Born of the same parents Pseudomonas and Ochrobactrum, wherein, Bacillus S3 has highest alkane (41.3%) and aromatic hydrocarbons (30.9%) degraded Rate.
Although bioanalysis, which administers oil polluted environment, substantial amounts of research, it is applied at present at oil pollution Most of degradation bacteria of reason comes from soil, and source Yu Haiyang habitat degradation bacteria report is relatively fewer.Corresponding to soil environment For, marine environment has the characteristics of low temperature, hypoxemia, oligotrophic, and launching high efficiency degradation bacteria often needs with dispensing nutrient source Deng, and can inoculating microbe retain its competitive edge for a long time to indigenous microorganism, if autochthons can be threatened, launched Whether nutriment can impact to ecological environment, and many scholars have different opinions to this.And screen ocean habitat degradation bacteria and exist Largely solve these problems.Therefore, turned into current by screening marine microorganism to administer petroleum pollution in ocean The direction of research.
The content of the invention
To solve the problems, such as petroleum pollution in ocean processing in the prior art, inventor passes through to be obtained to the screening of oil pollution marine site There must be the microbial strains of oil degradation performance, determine its classification position and its oil degradation performance is evaluated, simultaneously Research finds that bacterial strain P51 also has the function that degraded is luxuriant and rich with fragrance, established for degrading polycyclic aromatic hydrocarbons pollutant while degraded oil Basis.
To achieve the above object, the present invention adopts the following technical scheme that：
One plant have oil degradation function bacterial strain P51, Latin name be Tessaracoccus flavescens. its 16S rRNA sequences are as shown in SEQ ID No.1.Described bacterial strain P51 is collected in Talien New Port oil pollution marine site sea-bottom deposit Thing, concentration and separation gained.
Described bacterial strain P51 has been filed on preservation, and specific preservation information is as follows：
Depositary institution's title：China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC)；
Depositary institution address：The institute 3 of Chaoyang District, Beijing City Beichen Lu 1, Institute of Microorganism, Academia Sinica；
Preservation date：On July 12nd, 2017；
Deposit number：CGMCC No.14411；
Described bacterial strain P51 form and physicochemical characteristicses be：
Bacterial strain P51 is on solid LB media in 28 DEG C of condition underscore cultures to single bacterium colony, observation bacterial strain P51 bacterium colony Form, phage surface smooth bumps, how rounded neat in edge is, opaque, orange-yellow.
Bacterial strain P51 bacterial strains are in rod-short, atrichia, are about 0.74-0.84 μm, wide about 0.30-0.37 μm.
The separation method of the bacterial strain P51 comprises the following steps：Bacterial strain P51 is seeded to enrichment according to 1% inoculum concentration In culture medium, cultivated under the conditions of 15 DEG C, bacterial strain P51 bacteria suspension concentration is 108Cfu/ml, described enriched medium composition It is：It is added to using oil as sole carbon source in minimal medium, oil addition is the 0.5% of minimal medium total amount (v/v)。
Described minimal medium forms：MgSO4·7H2O 0.7g, NH4NO31g, KCl 0.7g, KH2PO42g, Na2HPO43g, natural sea-water 1000mL, PH 7.5,10mL micro-mixed liquor is added after sterilizing.
Above-mentioned bacterial strains P51 application is claimed in second purpose of the invention, i.e., in a manner of oil degradation and luxuriant and rich with fragrance degraded Carry out the improvement of oil pollution.
The method that above-mentioned bacterial strains P51 degraded oils are claimed in 3rd purpose of the invention, it is specially：
It is 10 by bacteria suspension concentration8Cfu/ml bacterial strains P51 is seeded in enriched medium according to 1% inoculum concentration, in 15 DEG C, 150r/min shaking tables concussion 15d.
The luxuriant and rich with fragrance method of above-mentioned bacterial strains P51 degradeds is：It is 10 by bacteria suspension concentration8Cfu/ml bacterial strains P51 according to 1% inoculation Amount is seeded in enriched medium, in 15 DEG C of quiescent culture 7d.
Compared with prior art, the beneficial effects of the invention are as follows：The present invention filters out one plant from bottom sediment first There is oil degradation and the bacterial strain P51 of luxuriant and rich with fragrance degraded simultaneously.After 15 DEG C of shaking tables shake 15d, petroleum degradation rate can reach the bacterial strain 28.90%.Tested and analyzed through GC-MS, bacterial strain P51 can reach 61.60% after 15 DEG C of quiescent culture 7d, to the degradation rate of phenanthrene. Found during the bacterial strain screening, bacterial strain P51 also has higher degraded while degraded oil, to polycyclic aromatic hydrocarbon China and Philippines Property, this characteristic solves caused marine pollution matter after oil pollution, marine environment is recovered completely, and is that exploitation is micro- Biological prosthetic oil pollution product lays the foundation.
Brief description of the drawings
Fig. 1 is bacterial strain P51 colonial morphology photo；
Fig. 2 is bacterial strain P51 thalli morphology photo；
Fig. 3 is bacterial strain P51 16SrRNA phylogenetic trees；
Fig. 4 is degraded photos of the bacterial strain P51 to oil；
Fig. 5 is degraded photos of the bacterial strain P51 to phenanthrene.
Embodiment
Technical scheme is further described with reference to instantiation, but the present invention is not in any form It is limited to embodiment content.Experimental method described in embodiment is conventional method unless otherwise specified；Unless otherwise specified, The reagent and biomaterial, are commercially obtained.
Embodiment 1
Bacterial strain P51 collection and separation
In Talien New Port oil pollution marine site bottom sediment, ice chest preserves sample collection after collection, and transports reality back rapidly Test the concentration and separation that room carries out oil degradation microorganism.
The freshly harvested sediment samples of about 10g are weighed to be added in the 250mL triangular flasks in enriched medium containing 100mL, 7d is enriched with the conditions of 15 DEG C of quiescent cultures, is then separated with gradient dilution partition method and obtains oil degradation bacterial strain.
Culture medium forms：
Minimal medium：MgSO4·7H2O 0.7g, NH4NO31g, KCl 0.7g, KH2PO42g, Na2HPO43g, day Right seawater 1000mL, PH 7.5,10mL micro-mixed liquor is added after sterilizing.
Micro-mixed liquor：CaCl22mg, FeCl3·6H2O 50mg, CuSO40.5mg, MnCl2·4H2O 0.5mg, ZnSO4·7H2O 10mg, distilled water 1000mL.
Enriched medium：In above-mentioned minimal medium, addition oil presses inorganic salts culture as sole carbon source, oil 1.5% (v/v) additions of base total amount.
Isolation medium：The agar solidification agent of 1.5% (mass percent) is added in enriched medium.
Embodiment 2
The morphologic observation and identification of bacterial strain：
Bacterial strain P51 is drawn on the enriched medium and LB culture mediums using oil as sole carbon source under the conditions of 15 DEG C respectively Line culture is to single bacterium colony, observation bacterial strain P51 colonial morphology.Picking colony fixes 1-2h with 2.5% glutaraldehyde solution simultaneously, so Bacteria suspension is dripped on silicon chip, drying in the air naturally to micro- wet leather hard afterwards, then silicon chip is rinsed with the phosphate buffer that PH is 7.2 10min, after rinsing 3 times, respectively with 30%, 50%, 70%, 85%, 95%, 100% Gradient elution using ethanol, then it is added dropwise to silicon chip Isoamyl acetate is fixed, and stands overnight.After the sample coating prepared, electron microscope sight will be scanned within second day Examine.As shown in figure 1, bacterial strain P51, on solid LB media, phage surface smooth bumps, how rounded neat in edge is, impermeable It is bright, in orange-yellow.As shown in Fig. 2 bacterial strain P51 is in rod-short, atrichia is about 0.74-0.84 μm, wide about 0.30-0.37 μm.
Exclude to repeat bacterial strain according to strain morphology and cultural characteristic, the bacterial strain of acquisition is then subjected to 16SrRNA gene sequences Row analysis.The microbe genome DNA obtained using multistage microwave amplifier separation, with bacterial universal primers F27：5′- AGAGTTTGATCCT GGCTCAG-3 ', 5 '-TACCTTGTTACGAC TT-3 ' (synthesis of Shanghai life work), enter performing PCR amplification, The GeneBank databases that the sequencing result of pcr amplification product is submitted to VCBI are compared, and using Blast softwares and The microbial strains phylogenetic tree that MEGA software buildings obtain, carry out Bacterial diversity population diversity analysis.Such as Fig. 3 institutes Bacterial strain P51 16S rRNA sequence analyses are shown as, with reference to its Observation On The Morphology, it is Tessaracoccus to identify bacterial strain P51 flavescens。
Embodiment 3
The measure of bacterial strain P51 oil degradation performances
Bacterial strain P51 oil degradations performance evaluation determines the petroleum degradation rate of bacterial strain using ultraviolet spectrometry indexing.With extraction The petroleum ether mixtures gone out are sample, and the oil content before and after strains for degrading is changed using U-5100 type ultraviolet spectrometry protractors Analyzed.Bacterial strain is to the degradation rate calculation formula of oil：η=(n0-n1)/n0× 100%, wherein η are petroleum degradation rate, n0For blank control, n1To be vaccinated with oil content remaining in the extract of the nutrient solution of bacterial strain.
150 μ L oil, which are added, in enriched medium carries out bacterial strain oil degradation performance evaluation as sole carbon source.By bacterial strain (bacteria suspension concentration is 108Cfu/ml) it is seeded to according to 1% inoculum concentration in enriched medium containing 100ml, it is quiet at 15 DEG C respectively Put after shaking 15d with 150r/min shaking tables, extracted with 300ml petroleum ethers, extract petrochina hydrocarbon is determined using GC-MS methods The degraded situation of each component.Not add the enriched medium of microbial strains to be repeated 3 times as control, experiment.
Under the conditions of 15 DEG C of shaking table cultures, bacterial strain P51 is to the degradation rate highest of oil, and 15d is up to 48.90%；15 DEG C quiet Put under condition of culture, bacterial strain P51 degradation rate is then significantly lower than shaking table culture, and 15d degradation rates are only 41.32%.Illustrate bacterial strain Degradeds of the P51 to oil in shaken cultivation, its degradation capability apparently higher than quiescent culture, oxygen in the degraded of oil very It is necessary.
Embodiment 4
The measure of bacterial strain P51 phenanthrene degradation properties
The evaluation of bacterial strain P51 phenanthrene degradation property determines the petroleum degradation rate of bacterial strain using gas chromatography.With extract two Chloromethane mixture is sample, using GC-MS-QP2010 types gas chromatograph-mass spectrometer (GC-MS) to luxuriant and rich with fragrance before and after strains for degrading Changes of contents is analyzed.Chromatographic column model HP-5MS (30m × 0.25nm × 0.25 μm), sample size are 1 μ L.Bacterial strain is to phenanthrene Degradation rate calculation formula be：η=(n0-n1)/n0× 100%, wherein η are luxuriant and rich with fragrance degradation rate, n0For the peak face of blank control group Product, n1For the experimental group peak area of inoculating strain.
150 μ L oil are added into enriched medium and carry out bacterial strain oil degradation performance evaluation as sole carbon source.By bacterium (bacteria suspension concentration is 10 for strain8Cfu/ml) it is seeded to according to 1% inoculum concentration in enriched medium containing 100ml, respectively at 15 DEG C Stand with after 150r/min shaking tables concussion 7d, extracted with 30ml dichloromethane, extract China and Philippines are determined using GC-MS methods Degraded situation.Not add the enriched medium of microbial strains to be repeated 3 times as control, experiment.
GC-MS analyzes luxuriant and rich with fragrance chromatographic condition：200 DEG C of injector temperature；Carrier gas is helium, and post case initial temperature is 40 DEG C, heating schedule is set as 40 DEG C of holding 5min, is warming up to 230 DEG C with 10.00 DEG C/min speed afterwards, keeps 17min；From Source temperature is 230 DEG C, scanning range 50-600amu.
When being cultivated under 15 DEG C of static conditions, bacterial strain P51 is to the degradation rate highest of phenanthrene, and 7d is up to 61.60%；15 DEG C of shaking tables Under condition of culture, bacterial strain P51 degradation rate is then significantly lower than quiescent culture, and 7d degradation rates are only 52.62%.Illustrate bacterial strain P51 In hypoxemia culture, its degradation capability is apparently higher than aerobic concussion and cultivate, and therefore, it is low that the bacterial strain is applied to seabed for degraded to phenanthrene The luxuriant and rich with fragrance degraded of warm low-oxygen environment.
By evaluating the degradation property of oil and phenanthrene bacterial strain P51, it is found that bacterial strain P51 has to oil and phenanthrene Preferable degradation property.Wherein, 15d is cultivated under the conditions of 15 DEG C of shaking tables, bacterial strain P51 is 28.90% to the degradation rate of oil； 7d is cultivated under 15 DEG C of static conditions, bacterial strain P51 is 61.60% to the degradation rate of phenanthrene.
Bibliography
[1] the former separation for knowing gorgeous littoral zone oil degradation bacteria and diversity analysis [D] Shandong：Qingdao Technological University, 2015：46-47
[2] Guo Qianyu, Zhang Jianmin, Li Rongrong, the screening of oiliness sewage treatment petrochina degradation bacteria and its degradation condition are excellent Change [J] Textile Colleges basic science journals, 2016,29 (2)：269-274
[3] Wang Jianan, Shi Yanyun, Zheng Liyan, the separation identification of oil degradation bacteria and 4 plants of bacillus interspecies effect [J] Environmental science, 2016,36 (6)：2245-2251.
Sequence table
SEQ ID No.1 (bacterial strain P51 (Tessaracoccus flavescens) 16S rRNA sequences)：
CGGGTGTTACCGACTTTCATGACTTGACGGGCGGTGTGTACAAGCCCCGGGAACGTATTCACCGCAGCGTTGCTGAT CTGCGATTACTAGCGACTCCGACTTCATGGGGTCGAGTTGCAGACCCCAATCCGAACTGAGACCGGCTTTCTGAGAT TCGCTCACCCTCACAGGCTCGCAGCTCTTTGTACCGGCCATTGTAGCATGCGTGAAGCCCTGGACATAAGGGGCATG ATGACTTGACGTCATCCCCACCTTCCTCCGAGTTGACCCCGGCGGTCTCCAATGAGTCCCCGGCATTACCCGCTGGC AACATTGGACGAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAGCCATGCACC ACCTGTATACCGACCAAAAAGGGGCACCTATCTCTAGATGTTTCCGGCATATGTCAAACCCAGGTAAGGTTCTTCGC GTTGCATCGAATTAATCCGCATGCTCCGCCGCTTGTGCGGGGCCCCGTCAATTCCTTTGAGTTTTAGCCTTGCGGCC GTACTCCCCAGGCGGGGTACTTAATGCGTTAGCTGCGGCACGGAGAACGTGGAATGTCCCCCACACCTAGTACCCAC CGTTTACGGCGTGGACTACCAGGGTATCTAAGCCTGTTTGCTCCCCACGCTTTCGCTTCTCAGCGTCAGGAAAGGTC CAGAGATCCGCCTTCGCCACCGGTGTTCCTCCTGATATCTGCGCATTCCACCGCTCCACCAGGAATTCCGATCTCCC CTACCTTCCTCAAGTCTGCCCGTATCGGAAGCAGGCTCAGTGTTGAGCACTGAGTTTTCACTCCCGACGTGACAAAC CGCCTACAAGCTCTTTACGCCCAATAAATCCGGACAACGCTCGCACCCTACGTATCACCGCGGCTGCTGGCACGTAG TTAGCCGGTGCTTCTTCTCCCACTACCGTCACGTTAGCTTCGTCATGGGTGAAAGCGGTTTACAACCCGAAGGCCGT CATCCCGCACGCGGCGTTGCTGCATCAGGCTTTCGCCCATTGTGCAATATTCCCCACTGCTGCCTCCCGTAGGAGTT TGGGCCGTATCTCAGTCCCAATGTGGCCGGTCGCCCTCTCAGGCCGGCTACCCGTCGAAGCCTTGGTGAGCCATTAC CTCACCAACAAGCTGATAGGCCGCGAGTCCATCCATGACCGCCGGAGCTTTCCAACCCCAAACATGAGTCCAGGGTT CATATCCGGTATTAGCACCTGTTTCCAGATGTTATCCCAGAGTCAAGGGCAGGTTACTCACGTGTTACTCACCCGTT CGCCACTCGTG。

Claims (7)

1. one plant of oil and phenanthrene degradation bacteria strain P51 for being applied to marine low temperature environment, it is characterised in that deposit number CGMCC No.14411, its 16SrRNA sequence such as SEQ ID No：Shown in 1.
2. a kind of separation method of bacterial strain P51 as claimed in claim 1, it is characterised in that comprise the following steps：By bacterial strain P51 It is seeded in enriched medium according to 1% inoculum concentration, is cultivated under the conditions of 15 DEG C, bacterial strain P51 bacteria suspension concentration is 108Cfu/ml, described enriched medium composition are：It is added to using oil as sole carbon source in minimal medium, oil Addition is 0.5% (v/v) of minimal medium total amount.
3. according to the method for claim 2, it is characterised in that described minimal medium, which forms, is：MgSO4·7H2O 0.7g, NH4NO31g, KCl 0.7g, KH2PO42g, Na2HPO43g, natural sea-water 1000mL, PH 7.5,10mL is added after sterilizing Micro-mixed liquor.
4. according to the method for claim 2, it is characterised in that described micro-mixed liquor：CaCl22mg, FeCl3· 6H2O 50mg, CuSO40.5mg, MnCl2·4H2O 0.5mg, ZnSO4·7H2O 10mg, distilled water 1000mL.
5. applications of the bacterial strain P51 as claimed in claim 1 in petroleum pollution.
6. application according to claim 5, it is characterised in that the method for bacterial strain P51 degraded oils is：By bacteria suspension concentration For 108Cfu/ml bacterial strains P51 is seeded in enriched medium according to 1% inoculum concentration, and in 15 DEG C, 150r/min shaking tables shake 15d。
7. application according to claim 5, it is characterised in that the luxuriant and rich with fragrance method of above-mentioned bacterial strains P51 degradeds is：Bacteria suspension is dense Spend for 108Cfu/ml bacterial strains P51 is seeded in enriched medium according to 1% inoculum concentration, in 15 DEG C of quiescent culture 7d.
CN201711067118.6A 2017-10-30 2017-10-30 Petroleum and phenanthrene degrading strain P51 suitable for seabed low-temperature environment Active CN107893037B (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
CN201711067118.6A CN107893037B (en) 2017-10-30 2017-10-30 Petroleum and phenanthrene degrading strain P51 suitable for seabed low-temperature environment
CN201711358029.7A CN108046441B (en) 2017-10-30 2017-10-30 Method for degrading petroleum and phenanthrene by using strain P51

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201711067118.6A CN107893037B (en) 2017-10-30 2017-10-30 Petroleum and phenanthrene degrading strain P51 suitable for seabed low-temperature environment

Related Child Applications (1)

Application Number Title Priority Date Filing Date
CN201711358029.7A Division CN108046441B (en) 2017-10-30 2017-10-30 Method for degrading petroleum and phenanthrene by using strain P51

Publications (2)

Publication Number Publication Date
CN107893037A true CN107893037A (en) 2018-04-10
CN107893037B CN107893037B (en) 2020-05-08

Family

ID=61803002

Family Applications (2)

Application Number Title Priority Date Filing Date
CN201711358029.7A Active CN108046441B (en) 2017-10-30 2017-10-30 Method for degrading petroleum and phenanthrene by using strain P51
CN201711067118.6A Active CN107893037B (en) 2017-10-30 2017-10-30 Petroleum and phenanthrene degrading strain P51 suitable for seabed low-temperature environment

Family Applications Before (1)

Application Number Title Priority Date Filing Date
CN201711358029.7A Active CN108046441B (en) 2017-10-30 2017-10-30 Method for degrading petroleum and phenanthrene by using strain P51

Country Status (1)

Country Link
CN (2) CN108046441B (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108949634A (en) * 2018-08-08 2018-12-07 东南大学 The oil degradation bacteria and its separation method of a kind of degradable heavy crude and application
CN113493752A (en) * 2021-07-15 2021-10-12 长江大学 Iron reducing bacterium Tessaracococcus oleiagri DH10 strain and application thereof

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110117559B (en) * 2019-04-29 2022-12-02 北京润世能源技术有限公司 Microbacterium W-Y13 and application thereof
CN113249272B (en) * 2021-06-28 2022-05-27 中国科学院水生生物研究所 Novel marine-source halotolerant zoogloea forming strain and application thereof

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106190923B (en) * 2016-08-11 2020-02-14 中山大学 Application of sphingolipid bacterium (Novosphingobium sp.)1MP25 strain in degrading polycyclic aromatic hydrocarbon
CN106315868A (en) * 2016-10-25 2017-01-11 山东大学 Application of degrading bacterium capable of metabolizing various hydrocarbons to petroleum pollutant disposal

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
CAI M等: "《Salinarimonas ramus sp. nov. and Tessaracoccus oleiagri sp. nov.,isolated from a crude oil-contaminated saline soil.》", 《INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY 》 *
CHEN, CHAO; LIU, QIU; LIU, CHANGJIAN; 等.: "《Effect of different enrichment strategies on microbial community structure in petroleum-contaminated marine sediment in Dalian, China》", 《MARINE POLLUTION BULLETIN 》 *
MARSH, N. A.等: "《Characterization of aliphatic hydrocarbon degrading anaerobes isolated from sand dune soils》", 《ABSTRACTS OF THE GENERAL MEETING OF THE AMERICAN SOCIETY FOR MICROBIOLOGY》 *
程之光主编: "《重油加工技术》", 30 June 1994 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108949634A (en) * 2018-08-08 2018-12-07 东南大学 The oil degradation bacteria and its separation method of a kind of degradable heavy crude and application
CN108949634B (en) * 2018-08-08 2022-03-08 东南大学 Petroleum degrading bacteria capable of degrading heavy crude oil and separation method and application thereof
CN113493752A (en) * 2021-07-15 2021-10-12 长江大学 Iron reducing bacterium Tessaracococcus oleiagri DH10 strain and application thereof
CN113493752B (en) * 2021-07-15 2022-05-06 长江大学 Iron reducing bacterium Tessaracococcus oleiagri DH10 strain and application thereof

Also Published As

Publication number Publication date
CN108046441A (en) 2018-05-18
CN107893037B (en) 2020-05-08
CN108046441B (en) 2020-06-23

Similar Documents

Publication Publication Date Title
AI-Jawhari Ability of some soil fungi in biodegradation of petroleum hydrocarbon
Bao et al. Biodegradation of crude oil using an efficient microbial consortium in a simulated marine environment
Hassanshahian Isolation and characterization of biosurfactant producing bacteria from Persian Gulf (Bushehr provenance)
CN107893037A (en) Oil and phenanthrene degradation bacteria strain P51 suitable for subsea cryogenic environment
Xia et al. Comparative study of biosurfactant produced by microorganisms isolated from formation water of petroleum reservoir
Morales-Guzmán et al. Diesel degradation by emulsifying bacteria isolated from soils polluted with weathered petroleum hydrocarbons
CN104745506A (en) Petroleum hydrocarbon degrading bacteria and application thereof
CN103215204A (en) Arthrobacter strain highly effectively degrading phenanthrene, and application thereof
CN105861388A (en) Oil field thermophilic degradation long chain hydrocarbon bacillus HNMC 11117 and application thereof
CN104388328B (en) Degrade bacterial strain and its acquisition methods, the application of 5 rings and 6 ring polycyclic aromatic hydrocarbons
CN1223673C (en) Erythro micrococcus Em and usage for generating biologic emulsifier as well as degrading polycyclic aromatic hydrocarbon
Fakhrzadegan et al. A study of crude oil‐degrading bacteria from mangrove forests in the Persian Gulf
CN105695360B (en) A kind of phenanthrene degradation bacteria Acinetobacter tandoii LJ-5 and its application
CN102399719A (en) Bacterium DW3 capable of degrading marine diesel oil pollutants
CN101935631B (en) Ralstoniasp. and application thereof in bioremediation of petroleum-contaminated saline-alkali soil
CN111733098A (en) Application of bacillus in low-temperature degradation of petroleum hydrocarbon
CN108034626A (en) A kind of degradation bacteria strains JN1 of oily sludge petrochina hydro carbons and its application
Baruah et al. Native hydrocarbonoclastic bacteria and hydrocarbon mineralization processes
CN106434413B (en) One kind is planted raw Raoul bacterium and the method using pyrene in bacterium degradation soil
Liu et al. Biodegradation and removal of heavy oil using Pseudomonas sp. and Bacillus spp. isolated from oily sludge and wastewater in Xinjiang Oilfield, China
CN105802883A (en) Method for improving screening efficiency of high-ring PAHs (polycyclic aromatic hydrocarbons) degrading bacteria
CN107629976A (en) A kind of microorganism and its microbial inoculum and application
CN106929454B (en) Strain CS07 with petroleum degradation and condensation performance and application thereof
CN108034625A (en) A kind of degradation bacteria strains JN7 of oily sludge petrochina hydro carbons and its application
Omotayo et al. Hydrocarbon degradation by free-living nitrogen-fixing bacteria

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant