CN1904033A - Diesel oil alkane eating bacteria and its degradation enzyme system - Google Patents
Diesel oil alkane eating bacteria and its degradation enzyme system Download PDFInfo
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- CN1904033A CN1904033A CN 200510085897 CN200510085897A CN1904033A CN 1904033 A CN1904033 A CN 1904033A CN 200510085897 CN200510085897 CN 200510085897 CN 200510085897 A CN200510085897 A CN 200510085897A CN 1904033 A CN1904033 A CN 1904033A
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- diesel oil
- eating bacteria
- alkane eating
- acid
- bacteria
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Abstract
The present invention provides an alcanivorax dieselolei NO1ACCTCC NO:M205024 obtained from deep sea. It can degrade straight-chain paraffin whose carbon chain length is 5C-36C.
Description
Technical field
The present invention relates to microbial technology field, specifically relate to a novel species alkane eating bacteria.
Background technology
Diesel oil is the oil fuel of various steamers, the mixture that it mainly is made up of straight-chain paraffin, branched paraffin, naphthenic hydrocarbon and a spot of aromatic hydrocarbon.Each state-owned nearly 80,000 ocean-going merchantman of the whole world shuttle back and forth in each harbour, the whole world at present, and they can reach more than 1,000,000 tons at the greasy dirt of term of voyage to ocean disposal every year.In addition, the marine oil overflow accident also frequently takes place in various places in recent years, marine oil pollution serious threat to the safety of the marine eco-environment.
Marine oil spill will experience dissolving, volatilization, photochemical oxidation decomposition, absorption and a series of processes such as sedimentation and biological degradation.Wherein, biological degradation is thoroughly to eliminate the most important approach of smeary, and the bioremediation technology that utilizes microbiological deterioration initiatively to remove oil spilling is current a kind of very important Environmental Biotechnology.Wherein, alkane eating bacteria is the very important oil degradation dominant bacteria of a class in the ocean.Before the present invention came forth, still nobody disclosed the diesel oil alkane eating bacteria that relates among the application.The present invention may provide important bacterial classification for petroleum hydrocarbon pollutes biological restoration.
Summary of the invention
The invention provides a kind of new microorganism-diesel oil alkane eating bacteria that petroleum hydrocarbon pollutes biological restoration that can be used in.Its called after diesel oil alkane eating bacteria Alcanivorax dieselollei (die.sel.o ' le.i, N.L.masc.n.dieseliuslatinized family name of Rudolf Diesel, L.gen.neut.n.olei of oil, N.L.gen neut.n.dieselolei of diesel oil, the sole carbon source used in the isolation procedure of thisstrain).Its preservation registration number is CCTCC NO:M205024.
Diesel oil alkane eating fungus strain of the present invention is by the bed mud collection of Eastern Pacific pelagite district website A (7 ° of 13 ' 46 " N, 153 ° 52 ' 19 " W, 5027 meters dark), and separation and Culture obtains in the laboratory.
Microorganism of the present invention is that 0.8-2.0 μ m is long, 0.3-0.7 μ m is wide, the flagellum of growing thickly, asporogenous Pueraria lobota Lan Shi negative bacillus.In NaCl concentration be on 3% the LB nutrient agar 28 ℃ cultivate and can form distinctive colonial morphology in 3 days: small, colourless, translucent, central protrusion, edge have irregular and transparent halo.Tween enzyme, katalaze enzyme and oxidase positive, esterase, arginine dihydrolase, amylase and gelatinase feminine gender.Can become nitrite to nitrate reduction.To the PXB sensitivity.Can not utilize carbohydrate.Can utilize following carbon source: polysorbate40, tween 80, methyl-prop ketone acid, monomethyl succsinic acid, p-oxyacetic acid phenyl ester, acetate, citric acid, lactic acid, beta-hydroxy-butanoic acid, gamma-hydroxybutyric acid, propionic acid and 2,3-butyleneglycol.Carbon source that can fine utilization has: alkane, citric acid, p-oxyacetic acid phenyl ester, pyruvic acid and lactic acid.Can utilize the straight-chain paraffin of carbon chain lengths 5C～36C, growth rapidly in 8C～28C.Main lipid acid is: C16:0, C18:1 ω 7c, C16: ω 7c, C19:0 cyclo ω 8c, C12:0 and C12:0 3OH.
Table 1 is diesel oil alkane eating bacteria of the present invention and the existing difference that belongs to other kind together.
Description of drawings
Fig. 1 is the transmission electron microscope photo of diesel oil alkane eating bacteria of the present invention.In lower part of figure, what arrow was indicated is the flagellum of growing thickly.
Embodiment
The following example is intended to further illustrate the present invention, rather than to the qualification of the present invention's scope.
Embodiment
The separation of diesel oil alkane eating bacteria NO1A
Gather deep-sea bed mud veneer of soil sample from Eastern Pacific's tuberculosis district website A, in the laboratory, can obtain diesel oil alkane eating bacteria NO1A of the present invention by following condition cultivation.
Artificial seawater substratum: NaCl=24g/L; MgSO
47H
2O=7.0g/L; NH
4NO
3=1g/L; KCl=0.7g/L; KH
2PO
4=2.0g/L; Na
2HPO
4=3.0g/L, pH7.4 adds an amount of micro-mixed solution after the sterilization, and mends with diesel oil as sole carbon source.Diesel oil and trace element are through 0.22 μ M membrane filtration degerming.The 100mL artificial seawater that soil sample is inoculated in diesel oil content 1% (v/v) is cultivated concentrated.28 ℃, 200r/min shaking table were cultivated after 4～5 days, take out 1mL and change in the 100mL fresh culture from nutrient solution.After the enrichment like this 6 times,, cultivate separation, purifying list bacterium colony after 48 hours for 28 ℃ with final enriched substance gradient dilution, spread plate.
SEQ ID N0.1 is the 16S rDNA sequence of diesel oil alkane eating bacteria of the present invention.The one of ordinary skilled in the art is appreciated that, also may be separated to same substantially microorganism from other environmental sample.Among these 16S rDNA sequences and the SEQ ID No.1 nucleotide sequence homology reach at least 98% or the microorganism that reaches more than 70% of DNA-DNA hybridization dependency also should be considered as belonging within the scope of the invention.
Table 1
| Feature | Alcanivorax dieselolei | Alcanivorax borkumensis | Alcanivorax venustensis | Alcanivorax jadensis |
| GC content (mol%) mobility/flagellum mode NO 3 -Reduction ion demand organic acid utilization: citric acid butanedioic acid β-carbonyl glutaric acid alpha-hydroxybutyric acid beta-hydroxy-butanoic acid gamma-hydroxybutyric acid optimum temperature (℃) 4 ℃ of 45 ℃ of growths of growth maximum NaCl concentration (%) the suitableeest NaCl concentration (%) | 62.1+, grow thickly+Na + + + - - + + 28-35 - + 15.0 3.0-7.5 | 53-54-+multiple (Na +，Mg 2+) - - + + + - 25-30 - - 12.5 3.0-10.0 | 66.4+, extremely give birth to-multiple---ND ND+23-25+-20.0 3.0-10.0 | 63-64 - + Na + - - ND ND ND ND 30 - - 15.0 3.0 |
Sequence table
The information of SEQ ID No.1:
(1) sequence signature
(A) length: 1504bp
(B) type: nucleic acid
(C) topological framework: linearity
(2) molecule type: tDNA
(3) sequence description: SEQ ID No.1
1 AGAGTTTGAT CCTGGCTCAG ATTGAACGCT GGCGGCAGGC CTAACACATG CAAGTCGAGC
61 GGAAACGATG GGAGCTTGCT CCCAGGCGTC GAGCGGCGGA CGGGTGAGTA ACACGTGGGA
121 ATCTGCCCAT TAGTGGGGGA TAACTCGGGG AAACTCGAGC TAATACCGCA TAATCCCTAC
181 GGGGGAAAGC AGGGGATCTT CGGACCTTGC GCTGATGGAT GAGCCCGCGT CGGATTAGCT
241 TGTTGGTGGG GTAATGGCCC ACCAAGGCGA CGATCCGTAA CTGGTCTGAG AGGATGGCCA
301 GTCACACCGG GACTGAGACA CGGCCCGGAC TCCTACGGGA GGCAGCAGTG GGGAATCTTG
361 GACAATGGGC GCAAGCCTGA TCCAGCCATG CCGCGTGTGT GAAGAAGGCC TTCGGGTTGT
421 AAAGCACTTT CAGTAGGGAG GAAGGCTTTG GGCTAATACC CTGGAGTACT TGACGTTACC
481 TACAGAAGAA GCACCGGCTA ATTTCGTGCC AGCAGCCGCG GTAATACGAA AGGTGCGAGC
541 GTTAATCGGA ATTACTGGGC GTAAAGCGCG CGTAGGCGGT GTGTTAAGTC GGATGTGAAA
601 GCCCAGGGCT CAACCTTGGA ATTGCATCCG ATACTGGCAC GCTAGAGTGC AGTAGAGGGA
661 GGTGGAATTT CCGGTGTAGC GGTGAAATGC GTAGAGATCG GAAGGAACAC CAGTGGCGAA
721 GGCGGCCTCC TGGACTGACA CTGACGCTGA GGTGCGAAAG CGTGGGGAGC AAACAGGATT
781 AGATACCCTG GTAGTCCACG CTGTAAACGA TGTCTACTAG CCGTTGGGGT CCTTAGTGAC
841 TTTGGTGGCG CAGCTAACGC GATAAGTAGA CCGCCTGGGG AGTACGGCCG CAAGGTTAAA
901 ACTCAAATGA ATTGACGGGG GCCCGCACAA GCGGTGGAGC ATGTGGTTTA ATTCGATGCA
961 ACGCGAAGAA CCTTACAGGC CTTGACATCC TGCGAACTTT CTAGAGATAG ATTGGTGCCT
1021 TCGGGAACGC AGTGACAGGT GCTGCATGGC TGTCGTCAGC TCGTGTCGTG AGATGTTGGG
1081 TTAAGTCCCG TAACGAGCGC AACCCTTGTC CTTAGTTGCC AGCACTTCGG GTGGGAACTC
1141 TAGGGAGACT GCCGGTGACA AACCGGAGGA AGGTGGGGAC GACGTCAAGT CATCATGGCC
1201 CTTACGGCCT GGGCTACACA CGTGCTACAA TGGTTGGTAC AGAGGGTTGC GAAGTCGCGA
1261 GGCGGAGCTA ATCTCTCAAA GCCAATCGTA GTCCGGATTG GAGTCTGCAA CTCGACTCCA
1321 TGAAGTCGGA ATCGCTAGTA ATCGCGGATC AGAATGCCGC GGTGAATACG TTCCCGGGCC
1381 TTGTACACAC CGCCCGTCAC ACCATGGGAG TGGATTGCAC CAGAAGTAGT TAGTCTAACC
1441 TTCGGGAGGA CGATTACCAC GGTGTGGTTC ATGACTGGGG TGAAGTCGTA ACAAGGTAGC
1501 CGTA
Claims (2)
1, a kind of alkane alkane eating bacteria is characterized in that called after diesel oil alkane eating bacteria Alcanivorax dieselolei, and its preservation registration number is CCTCC NO:M205024.
2, diesel oil alkane eating bacteria as claimed in claim 1, it is characterized in that the nucleotides sequence among its 16S rDNA sequence and the SEQ IDNo.1 shows at least 98% homology, perhaps the DNA-DNA of the genomic dna of described diesel oil alkane eating bacteria and bacterial strain CCTCC NO:M205024 hybridization dependency reaches more than 70%.
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|---|---|---|---|
| CN 200510085897 CN1904033A (en) | 2005-07-20 | 2005-07-20 | Diesel oil alkane eating bacteria and its degradation enzyme system |
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|---|---|---|---|
| CN 200510085897 CN1904033A (en) | 2005-07-20 | 2005-07-20 | Diesel oil alkane eating bacteria and its degradation enzyme system |
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| Publication Number | Publication Date |
|---|---|
| CN1904033A true CN1904033A (en) | 2007-01-31 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104498379A (en) * | 2014-07-24 | 2015-04-08 | 国家海洋局第三海洋研究所 | Marine oil-degrading bacterium, fermentation compound and application thereof |
| CN109337938A (en) * | 2018-08-31 | 2019-02-15 | 河南师范大学 | Utilize the method for alkane eating bacteria fermentative degradation paraffin production 2- oxo adipic acid |
| CN115747124A (en) * | 2023-01-09 | 2023-03-07 | 海南热带海洋学院 | Alkylophagus, application and culture method thereof and method for degrading plastics |
-
2005
- 2005-07-20 CN CN 200510085897 patent/CN1904033A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104498379A (en) * | 2014-07-24 | 2015-04-08 | 国家海洋局第三海洋研究所 | Marine oil-degrading bacterium, fermentation compound and application thereof |
| WO2016011805A1 (en) * | 2014-07-24 | 2016-01-28 | 国家海洋局第三海洋研究所 | Marine oil-degrading bacterium, fermentation compound and uses thereof |
| CN104498379B (en) * | 2014-07-24 | 2017-11-14 | 国家海洋局第三海洋研究所 | One plant of offshore oil degradation bacteria, fermentation gained compound and application thereof |
| US9920387B2 (en) | 2014-07-24 | 2018-03-20 | Third Institute Of Oceanography, State Oceanic Administration | Strain of marine oil-degrading bacteria, compounds obtained by fermentation and their applications |
| CN109337938A (en) * | 2018-08-31 | 2019-02-15 | 河南师范大学 | Utilize the method for alkane eating bacteria fermentative degradation paraffin production 2- oxo adipic acid |
| CN109337938B (en) * | 2018-08-31 | 2021-05-07 | 河南师范大学 | Method for producing 2-oxoadipic acid by fermenting and degrading paraffin with alcanivorax |
| CN115747124A (en) * | 2023-01-09 | 2023-03-07 | 海南热带海洋学院 | Alkylophagus, application and culture method thereof and method for degrading plastics |
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