CN104004686A - Polycyclic aromatic hydrocarbons degrading bacterium s21-t17(H)-N3 and application thereof - Google Patents
Polycyclic aromatic hydrocarbons degrading bacterium s21-t17(H)-N3 and application thereof Download PDFInfo
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- CN104004686A CN104004686A CN201410248117.1A CN201410248117A CN104004686A CN 104004686 A CN104004686 A CN 104004686A CN 201410248117 A CN201410248117 A CN 201410248117A CN 104004686 A CN104004686 A CN 104004686A
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- polycyclic aromatic
- aromatic hydrocarbons
- erythrobacter
- aromatic hydrocarbon
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Abstract
The invention provides a polycyclic aromatic hydrocarbons degrading bacterium s21-t17(H)-N3 and application thereof and relates to biological treatment of environmental pollutants. The polycyclic aromatic hydrocarbons degrading bacterium (Erythrobacter sp.) s21-t17(H)-N3 is registered with the number of CGMCC No.9030 and can be applied to degradation of various organic compounds. The organic compounds are polycyclic aromatic hydrocarbon compounds, and the polycyclic aromatic hydrocarbon compounds comprise but not limited to naphthalene, phenanthrene, anthracene, fluoranthene, chrysene, pyrene, benzopyrene and the like. Genomic sequencing is carried out on the polycyclic aromatic hydrocarbons degrading bacterium (Erythrobacter sp.)s21-t17(H)-N3, and results show that the polycyclic aromatic hydrocarbons degrading bacterium (Erythrobacter sp.)s21-t17(H)-N3 contains a dioxygenase gene related to polycyclic aromatic hydrocarbons. The dioxygenase gene plays an important role when the polycyclic aromatic hydrocarbons are degraded.
Description
Technical field
The present invention relates to environmental pollutant biological treatment, especially relate to a polycyclic aromatic hydrocarbon degrading bacteria s21-t17 (H)-N3 and application thereof.
Background technology
Polycyclic aromatic hydrocarbons (Polycyclic Aromatic Hydrocarbons, PAHs) is the aromatic compound that a class contains at least 1 phenyl ring, and chemical structure is very stable, and hydrophobicity is strong, is difficult for degraded, including naphthalene, phenanthrene, pyrene etc., has kind more than 200.PAHs has high toxicity, there is very strong " three cause " effect, that is: carcinogenic, teratogenesis and mutagenesis (Sato H, Aoki Y.Mutagenesis by environmental pollutants and bio-monitoring of environmental mutagens[J] .Curr Drug Metab.2002,3 (3): 311-319; Phillips DH.Fifty years of benzo (a) pyrene[J] .Nature.1983,303 (5917): 468-472; Sternberg SS.The carcinogenesis, mutagenesis and teratogenesis of insecticides.Review of studies in animals and man[J] .Pharmacology & Therapeutics.1979,6 (1): 147-166.).PAHs is a class maximum in environmental carcinogenesis chemical substance, wherein, benzo [α] pyrene, benzo [α] anthracene etc. are that harm is maximum, benzo [α] pyrene is first found environmental chemical carcinogen, be appointed as one-level carcinogenic substance by international cancer research institution, classified one of conventional project of home and abroad environment monitoring as.PAHs has been subject to the attention of various countries' environment protection tissue; Environmental Protection Agency is by 16 kinds of pollutent Black List (Autrup H that listed preferential improvement with substituent PAHs in; Harris CC; Trump BF et al.Metabolism of benzo (a) pyrene and identification of the major benzo (a) pyrene-DNA adducts in cultured human colon[J] .Cancer Res.1978,38 (11Pt1): 3689-3696.).
Most of PAHs in atmosphere derives from the incomplete combustion of coal and oil, is mainly derived from solid waste, industry is leaked and blended atmospheric precipitation in terrestrial environment.The main PAHs of ocean environment source is the former oil seepage in seabed, sea transport, oil well seepage, oil tanker accident etc.PAHs is the main ingredient except alkane in oil.Average every day is exploited out 70,000,000 gallons of oil in the whole world, wherein has half to transport by ocean, and oil well nature seepage, sea transport leakage etc. inevitably can occur in the production and transport process of oil.More seriously the large-scale pollution causing due to oil tanker accident, in pollutent, volatile composition enters in atmosphere by evaporation, indissoluble is solved problem the composition minority of volatilizing can be by approach degradeds such as photoxidation, majority by attach to oceanic sediment for years in ocean, accumulate, PAHs just belongs to the latter.Thereby the PAHs remaining in ocean may enrichment poison marine organisms in marine organism.More very be, the compound of this difficult degradation may directly have influence on human health (Shiaris MP through food chain cumulative function, D.J-S.Polycyclic aromatic hydrocarbons in surficial sediments of Boston Harbour, MA, USA.[J] .Mar Poll Bull.1986,17:469～472.; Baumard, Budzinski, Garrigues.Polycyclic aromatic hydrocarbons in sediments and mussels of the western Mediterranean Sea[J] .Environ Toxicol & Chem.1998,15:765-776.).
PAHs in environment can remove in several ways, comprise nature volatilization, photodegradation, chemical degradation, biological degradation, particle absorption etc., microbial transformation and degraded are considered to PAHs in environment and remove the most thoroughly efficient manner (Dua M, Singh, A., Sethunathan, N., Johri, A.K..Biotechnology and bioremediation:successes and limitations[J] .ApplMicrobiolBiotechnol.2002,59 (2-3): 143-152.).At present, in known environment, PAHs degrading microorganism has manyly, comprises bacterium, actinomycetes, fungi, algae etc.The existing research widely of degradation by bacteria of PAHs, the separated PAHs degradation bacteria that is accredited as of many bacteriums, these bacterial strains mainly comprise the genus such as Pseudomonas, Sphingomonas, Cycloclasticus, Burkholderia, Rhodococcus, Polaromonas, Neptunomonas, Janibacter, Nocardia, Bacillus, Mycobacterium.
Summary of the invention
The object of this invention is to provide a polycyclic aromatic hydrocarbon degrading bacteria (Erythrobacter sp.) s21-t17 (H)-N3.
Another object of the present invention is to provide the application of a polycyclic aromatic hydrocarbon degrading bacteria (Erythrobacter sp.) s21-t17 (H)-N3 in the various organic compound of degraded.
Described polycyclic aromatic hydrocarbon-degrading bacteria (Erythrobacter sp.) s21-t17 (H)-N3, in 04 Yue03 China Committee for Culture Collection of Microorganisms common micro-organisms center preservation in 2014, preservation address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode: 100101, register on the books and be numbered CGMCC No.9030.Described polycyclic aromatic hydrocarbon-degrading bacteria (Erythrobacter sp.) s21-t17 (H)-N3 derives from the deposit on marine-bottom surface of South Atlantic, obtains through artificial enrichment culture, separation and purification.This bacterium is that the red bacterium of Gram-negative belongs to bacterial strain s21-t17 (the H)-N3 of (Erythrobacter sp.), biological characteristics is catalase feminine gender, oxidase positive, non-fermented type, obligate is aerobic, thalli morphology is rounded, safran is opaque, regular edges, smooth surface projection, without halo, diameter 3～4mm.The optimum growing condition of this bacterium is: pH=6.5～8.5,25～28 DEG C of temperature.
Described polycyclic aromatic hydrocarbon-degrading bacteria (Erythrobacter sp.) s21-t17 (H)-N3 can be in the application of degrading in various organic compound.
Described organic compound is polycyclic arene compound, and described polycyclic arene compound includes but not limited to naphthalene, phenanthrene, and anthracene, fluoranthene, bends at (Fourth Ring) pyrene, the compounds such as benzopyrene.
Described polycyclic aromatic hydrocarbon-degrading bacteria (Erythrobacter sp.) s21-t17 (H)-N3 is carried out to gene order-checking discovery, and this bacterium contains the dioxygenase gene relevant to polycyclic aromatic hydrocarbons.These dioxygenase genes play a significant role in the time of this bacterium degrading polycyclic aromatic hydrocarbons.
Experiment showed, that described polycyclic aromatic hydrocarbon-degrading bacteria (Erythrobacter sp.) s21-t17 (H)-N3 can utilize mixing PAHs, naphthalene, phenanthrene, anthracene, fluoranthene, bend at (Fourth Ring), the polycyclic aromatic hydrocarbonss such as pyrene and diesel oil are as unique energy and carbon source for growth breeding, and 20 days energy of this bacterium are by the mixing PAHs in minimal medium, naphthalene, phenanthrene, anthracene, fluoranthene, bend at (Fourth Ring) pyrene, benzopyrene and Diesel degradation 67.24%, 68.26%, 71.54%, 39.44%, 35.12%, 22.14%, 27.34%, 25.46%, 50.47%.
Brief description of the drawings
Fig. 1 is the degradation rate of s21-t17 (H)-N3 in various carbon sources.
Embodiment
The performance that the separation of example one: Erythrobactersp.s21-t17 (H)-N3 mixes polycyclic aromatic hydrocarbons with degraded
The deposit on marine-bottom surface of collection in worksite South Atlantic, adopts 100mL sea water medium to add naphthalene, phenanthrene, pyrene mixing polycyclic aromatic hydrocarbons to carry out enrichment culture under 25 DEG C of 180r/min conditions.After two weeks, dilution spread flat board in solid medium (M2), obtain the pure bacterium of many strains, pure bacterium is through being seeded to the ability of verifying its decomposing petroleum hydrocarbon in liquid nutrient medium, finally obtain efficient degrading bacteria s21-t17 (the H)-N3 of a strain polycyclic aromatic hydrocarbons hydrocarbon, this bacterium is accredited as red bacterium through 16S rDNA and belongs to (Erythrobactersp.).The composition of solid medium (M2): sodium acetate 5g/L, Trisodium Citrate 0.05g/L, broth medium 0.5g/L, peptone 0.1g/L, yeast powder 0.1g/L, oxysuccinic acid 0.05g/L, glucose 1g/L, sucrose 0.5g/L, ammonium nitrate 1g/L, ammonium chloride 0.2g/L, sodium tartrate 0.05g/L, Zulkovsky starch 0.5g/L, agar powder 15g/L, add seawater to 1L, adjust pH7.5, after 115 DEG C of high pressure steam sterilization 30min, add the sterilized KH of 1% volume
2pO
4the FeSO of solution (10g/L, pH6.7) and 0.1% volume 0.22 μ m filter membrane degerming
4(0.4g/L).
Erythrobactersp.s21-t17 (H)-N3 is seeded in the artificial seawater substratum (MMC) containing naphthalene, phenanthrene, pyrene mixing polycyclic aromatic hydrocarbons, in 180r/min, 25 DEG C of shaking tables, cultivate after 20 days, measure this bacterium for the degradation capability that mixes polycyclic aromatic hydrocarbons, obtain through measuring, this bacterium is 67.24% (Fig. 1) to the degradation rate that contains naphthalene, phenanthrene, pyrene mixing polycyclic aromatic hydrocarbons.Artificial seawater substratum (MMC): NaCl24g/L, MgSO
47H
2o7g/L, NH
4nO
31g/L, KCl0.7g/L, KH
2pO
42g/L, Na
2hPO
43g/L, pH7.4.After sterilizing, add appropriate micro-mixed solution, trace element is through 0.22 μ M membrane filtration degerming.MMC trace element mixed solution: CaCl
2, 2mg/L; FeCl
36H
2o, 50mg/L; CuSO
4, 0.5mg/L; MnCl
24H
2o, 0.5mg/L; ZnSO
47H
2o, 10mg/L.
The degradation property of example two: Erythrobactersp.s21-t17 (H)-mono-polycyclic aromatic hydrocarbons of N3 is measured
Erythrobactersp.s21-t17 (H)-N3 is seeded to different carbon source (naphthalenes, luxuriant and rich with fragrance, anthracene, fluoranthene, bends at (Fourth Ring) pyrene, diesel oil) artificial seawater substratum (MMC) in, end blank (only add substrate and do not add bacterium) is set simultaneously, in 180r/min, 28 DEG C of shaking tables, cultivates and observe afterwards the upgrowth situation of bacterial strain in 20 days, and measure the degradation rate of this bacterium to each substrate.Result shows that this bacterial strain can utilize naphthalene, phenanthrene, and anthracene, fluoranthene, bends at (Fourth Ring) pyrene, benzopyrene, diesel oil growth, degradation rate is respectively 68.26%, 71.54%, and 39.44%, 35.12%, 22.14%, 27.34%, 25.46%, 50.47% (Fig. 1).
The application of example three: Erythrobactersp.s21-t17 (H)-N3 degrading polycyclic aromatic hydrocarbons genes involved
Erythrobactersp.s21-t17 (H)-N3 is carried out to gene order-checking, from genome, find and degrading polycyclic aromatic hydrocarbons genes involved, find many aromatic hydrocarbon degraded genes involveds (table 1);
Degrading polycyclic aromatic hydrocarbons genes involved in table 1 Erythrobactersp.s21-t17 (H)-N3 genome
Described gene can be applied in degrading polycyclic aromatic hydrocarbons.
Claims (7)
1. polycyclic aromatic hydrocarbon-degrading bacteria (Erythrobacter sp.) s21-t17 (H)-N3,, in 04 Yue03 China Committee for Culture Collection of Microorganisms common micro-organisms center preservation in 2014, register on the books and be numbered CGMCC No.9030.
2. the application of polycyclic aromatic hydrocarbon-degrading bacteria (Erythrobacter sp.) s21-t17 (H)-N3 in the various organic compound of degraded as claimed in claim 1.
3. application as claimed in claim 2, is characterized in that described organic compound is polycyclic arene compound.
4. application as claimed in claim 3, is characterized in that described polycyclic arene compound includes but not limited to naphthalene, phenanthrene, and anthracene, fluoranthene, bends at (Fourth Ring) pyrene, benzopyrene.
5. a polycyclic aromatic hydrocarbon degrading bacteria (Erythrobacter sp.) s21-t17 (H)-N3 as claimed in claim 1, is characterized in that it has the dioxygenase gene relevant to degrading polycyclic aromatic hydrocarbons;
Degrading polycyclic aromatic hydrocarbons genes involved in table 1 Erythrobactersp.s21-t17 (H)-N3 genome
Described gene is in table 1.
6. a polycyclic aromatic hydrocarbon degrading bacteria (Erythrobacter sp.) s21-t17 (H)-N3 as claimed in claim 5, is characterized in that described gene applies in polycyclic aromatic hydrocarbons contaminated environmental monitoring and biological restoration.
7. a polycyclic aromatic hydrocarbon degrading bacteria (Erythrobacter sp.) s21-t17 (H)-N3 as claimed in claim 5, is characterized in that described gene applies in degrading polycyclic aromatic hydrocarbons.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105624058A (en) * | 2015-12-04 | 2016-06-01 | 辽宁省海洋水产科学研究院 | Marine diesel oil degrading bacterial strain BHB-16 and immobilization method thereof |
| CN108504614A (en) * | 2018-03-28 | 2018-09-07 | 新疆大学 | A kind of degrading polycyclic aromatic hydrocarbons engineering bacteria wp4-C23O and its construction method and application |
| CN110656069A (en) * | 2019-11-07 | 2020-01-07 | 自然资源部第三海洋研究所 | Petroleum hydrocarbon degrading bacteria and culture, screening and application thereof |
| CN113577637A (en) * | 2021-09-07 | 2021-11-02 | 中国科学院南京土壤研究所 | Method for enhancing microbial degradation of polycyclic aromatic hydrocarbon and microbial agent |
| CN114703222A (en) * | 2022-03-15 | 2022-07-05 | 上海市农业科学院 | Cultivation method of completely degradable polycyclic aromatic hydrocarbon plant |
-
2014
- 2014-06-06 CN CN201410248117.1A patent/CN104004686A/en active Pending
Non-Patent Citations (2)
| Title |
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| 谭田丰: "海洋石油降解菌的分离鉴定及其在海洋石油污染生物修复中的应用研究", 《中国优秀硕士学位论文全文数据库工程科技I辑》 * |
| 谭田丰: "海洋石油降解菌的分离鉴定及其在海洋石油污染生物修复中的应用研究－2", 《中国优秀硕士学位论文全文数据库工程科技I辑》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105624058A (en) * | 2015-12-04 | 2016-06-01 | 辽宁省海洋水产科学研究院 | Marine diesel oil degrading bacterial strain BHB-16 and immobilization method thereof |
| CN108504614A (en) * | 2018-03-28 | 2018-09-07 | 新疆大学 | A kind of degrading polycyclic aromatic hydrocarbons engineering bacteria wp4-C23O and its construction method and application |
| CN110656069A (en) * | 2019-11-07 | 2020-01-07 | 自然资源部第三海洋研究所 | Petroleum hydrocarbon degrading bacteria and culture, screening and application thereof |
| CN113577637A (en) * | 2021-09-07 | 2021-11-02 | 中国科学院南京土壤研究所 | Method for enhancing microbial degradation of polycyclic aromatic hydrocarbon and microbial agent |
| CN114703222A (en) * | 2022-03-15 | 2022-07-05 | 上海市农业科学院 | Cultivation method of completely degradable polycyclic aromatic hydrocarbon plant |
| CN114703222B (en) * | 2022-03-15 | 2024-02-02 | 上海市农业科学院 | Cultivation method of plant capable of completely degrading polycyclic aromatic hydrocarbon |
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Application publication date: 20140827 |