CN105505827B - The gene and application of petroleum sweetening denitrogenation bacterial strain and its desulfurization removing nitric - Google Patents

The gene and application of petroleum sweetening denitrogenation bacterial strain and its desulfurization removing nitric Download PDF

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CN105505827B
CN105505827B CN201610012840.9A CN201610012840A CN105505827B CN 105505827 B CN105505827 B CN 105505827B CN 201610012840 A CN201610012840 A CN 201610012840A CN 105505827 B CN105505827 B CN 105505827B
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CN105505827A (en
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王威
朱琳
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Tianjin Weirui Biological Technology Co ltd
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Abstract

The present invention relates to the bacterial strains to oil with desulfurization removing nitric effect and its desulfurization removing nitric gene and application, entitled W 1 to be preserved in " China Committee for Culture Collection of Microorganisms's common micro-organisms center ", preserving number CGMCC 11780.The strain is isolated from oil field stratum water sample；The strain that the present invention identifies belongs to Soil Bacillus category, and high temperature resistant has good thermal stability, can be applied in the industrial production that fermentation etc. needs thermophilic enzyme condition；It can in reservoir media well-grown, many and element sulphur and nitrogen related gene in sour gas field in removing oil are found by carrying out genome sequencing to W 1, it is demonstrated experimentally that the bacterium has the ability of desulfurization removing nitric, it is remarkably improved the biological desulphurization of fuel oil（Nitrogen）Efficiency simultaneously effectively solves the problem of environmental pollution after use, has the good characteristic of commercial Application bacterial strain, bacterial strain has wide commercial application potentiality.

Description

The gene and application of petroleum sweetening denitrogenation bacterial strain and its desulfurization removing nitric
Technical field
The present invention relates to microbial strains and field of environment microorganism, and in particular to one plant of hot glucoside of petroleum sweetening denitrogenation Ground bacillus W-1 and its degrading genes and application.
Background technology
21 century is environmentally friendly century, and the energy industry as chemical industry main force all suffers from perhaps in environmental protection and process aspect More challenges and opportunity, and the new branch of science that be thus born biochemical industry and field of environment protection are intersected.With global industry The fast development of process increases severely to the demand of the energy, currently, world energy sources structure petrochina proportion is maximum, is 37.4%.There are sulphur, nitrogen compounds in oil product can cause environmental pollution using containing sulphur, nitrogen compound this product, right Equipment generates corrosion.Therefore, desulfurization, denitrogenation technology are the important subjects of petrochemical industry and environmental protection.
Oil and products thereof deep desulfuration is the important class that China or even whole world petrochemical industry are faced in the new century Topic.Sulphur in oil is mainly with mercaptan, thioether, thiophene, benzothiophene（BT）And their alkyl substituents and derivative etc. have The form of machine sulfide exists, these compounds generate a large amount of rhodanide in burning, bring a variety of harm.Wherein the most The serious is SO caused by burning2And SO3Acid rain easily is formed with vapor in the atmosphere chemical combination, causes serious pollution, is broken The bad ecological balance is detrimental to health（Monticello D J. desulfurization of fossil fuels.Ann Rev Microbiol.1985,39:371~389）.In recent years, as biotechnology desulfurization is increasingly that the people are of interest.Biology Catalytic desulfurization（BDS）It is that a kind of removed at normal temperatures and pressures using aerobic, anaerobic bacteria combines sulphur in oil sulfur heterocyclic compound A kind of new technology, sulphur in fossil fuel removed in such a way that biochemistry cracks by the distinctive enzyme of microorganism, is in room temperature Normal pressure and not needing carries out under conditions of hydrogen, can specificity cut-out carbon-sulfur bond and make the sulphur in fossil fuel with the shape of sulfate Formula releases, and the purpose is to utilize hydro carbons of the microorganism removing sulphur without destroying organic-fuel value existing for nature.Its Feature is that process costs are low, and saving energy, flow is simple, and reaction condition is mild, clean environment, meets the development of eco-friendly Trend.Decades have been developed from appearance in BDS technologies so far, so far still in the developmental research stage.Living things catalysis Desulfurization technology is the removal of organic sulphur component in oil, is opened up a new way.
Nitrogen content is generally lower than sulfur content in oil, and mass fraction is only a small number of former usually in 0.05% ~ 0.5% range The nitrogen content score of oil is more than 0.6%.Nitrogen content is higher in crude oil in China, and concentration about 90% in the residual oil of most of crude oil Nitrogen, therefore when processing crude oil in China, the nitrogen content in crude oil should be paid attention in again.The presence of organic nitrogen compound is to oil refining in crude oil Technique, oil product and environment have very big harm.Although the value volume and range of product of Nitrogen Compounds in Crude Oils does not have, sulfide is more, it Influence be greater than sulfide.In Oil Production, the organic nitrogen compound of denier can cause catalytic cracking, hydrofinishing etc. Valuable catalyst is poisoned in technical process, shortens the service life of catalyst, to increase production cost, reduces yield.According to Statistics reduces 90% of nitrogen content in crude oil, can improve 20% gasoline production, moreover it is possible to reduce equipment corrosion, reduce expense.Separately Outside, nitride can influence the color and stability of oil product, and especially during storage, the presence of nitride can cause oil colours to become Deep and generation colloid and precipitation;The burning of nitride can generate the very strong sour gas of corrosivity, with nitrogen oxides NOXForm It is discharged into air, causes serious air pollution, and form acid rain.Therefore, the nitrogen removed in oil is particularly important, and traditional Bio-denitrification technology haves the shortcomings that various very important：Of high cost, operation difficulty is big, removing substrate kind is limited, Removal effect is not notable, and the research report of existing related denitrification microorganism is almost all room temperature microorganism.
The bacterial strain of the present invention belongs to thermophilus strain, has the function of while to petroleum sweetening denitrogenation, can be applied to petrochemical industry In industry, environmental protection industry and the production of other related industries.
Invention content
The main purpose of the present invention is to provide a kind of hot glucoside ground bacillus W-1 (Geobacillus thermoglucosidasius) and its identify and apply, the strain of identification belongs to Soil Bacillus category, and high temperature resistant has Good thermal stability can be applied in the industrial production that fermentation etc. needs thermophilic enzyme condition；Oil can effectively be removed Middle sulfur-bearing（Nitrogen）Sulphur in compound（Nitrogen）And the bacterial strain is in sulfur-bearing（Nitrogen）Crude oil and its fraction desulfurization removing nitric in application.
Second object of the present invention is to provide the identification mark and 16S rRNA sequences of the bacterial strain.
Third object of the present invention is to provide the bacterial strain remove sulfur-bearing nitrogenous compound in element sulphur, nitrogen side Method.
The present invention is applied to Oil Generation object depth degree desulfurization removing nitric process.
Technical scheme of the present invention is summarized as follows:
Hot glucoside ground bacillus provided by the present invention, it is characterised in that：The bacterial strain isGeobacillus thermoglucosidasius, it is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms on December 4th, 2015 Center（China General Microbiological Cultuer Collection Centre）Progress preservation, preservation Location：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, postcode：100101, register into Volume number is CGMCC NO.11780.
（1）The bacterial strain isolates and purifies to obtain from the water sample of Dagang Oilfield stratum, is sequenced through full genome, 16S rDNA annotations Belong toGeobacillus thermoglucosidasius.The bacterium is gram-positive bacteria, and biological characteristics are catalase sun Property, volume morphing is to have the long bacillus of gemma, bacterium colony rounded, and milky is slightly swelled, smooth-shaped, neat in edge, moistening, impermeable It is bright；It can gelatin hydrolysate, starch and casein；NO3-N and NO2-N can be restored；Citrate cannot be utilized；Higher than 3% NaC l growths are suppressed.By the sequencing to the bacterium genome, it is found that it is much related to petroleum sweetening denitrogenation the bacterium contains Gene.The optimum growing condition of the bacterium is：60-65 DEG C of temperature, pH=7.0 ~ 7.5, colonial morphology such as Fig. 1.
（2）The bacterial strain is carried out with 50 CHB/E indentifying substances items of ABI to produce sour experiment using different carbon source, is as a result shown The bacterial strain can utilize ribose, D- xyloses, L-arabinose, glucose, mannose, fructose, mannitol, N-Acetyl-D-glucosamine, Cellobiose, maltose, sucrose, trehalose, scholar's logical sequence sugar, cannot utilize the fermentations such as glycerine, D-arabinose, sorbierite, lactose Production acid.
（3）By rightGeobacillus thermoglucosidasiusFull genome sequencing is carried out to find,Geobacillus thermoglucosidasiusRelated gene with element sulphur in many sulfur-bearing nitrogenous compounds with removing （Referring to table 1）, nitrogen related gene（Referring to table 2）, these genes are for the desulfuration in terms of petrochemical industry and environmental protection Denitrification is significant.
In 1 W-1 genomes of table with the relevant gene of desulfurization
The described and removing relevant gene of sulphur is referring to table 1
In 2 W-1 genomes of table with the relevant gene of denitrogenation
The described and removing relevant gene of nitrogen is referring to table 1
Nucleotide sequence SEQ ID No.2：
orf00188
ATGGAATTATTATGGTTTATCCCTTCCCATGGTGATGGGCGGTATCTTGGAACGACCAAAGGAGGACGC GCTCCGGAATACAGCTATTTCCGGCAAATCGCCCAAGCGGCTGACCGGCTTGGCTATAAAGGGGTATTGATTCCGAC AGGAAAATCGTGCGAAGATCCGTGGCTGCTTGCGTCAGCATTAGCGGCAGAAACCGAACAATTGCGTTTTCTCGTTG CCGTGCGACCGGGGTTAATGTCTCCTACGCTAGCAGCACGCATGGCTTCCACATTGGACCGCATTTCGGAAGGCCGG TTGCTTATCAATGTCGTAGCTGGCGGCGATCCTGTTGAGCTTGCGGGCGATGGGCTATTTTTAAGCCATGACGAACG TTATGAAGCGACGGATGAATTTTTAACCGTTTGGAAACGGCTGTTAAGCGGCGAAGAAGTCACTTTAAAAGGGAAAC ATATCCATGTGAATGAGGCAAAGCTTCTGTTTCCGCCAACTCAAAAACCGTATCCGCCGATTTATTTTGGCGGGTCG TCACCGGCAGGCCAACTTGTGGCAGCAAAACATGCGGATGTTTACTTAACATGGGGGGAACCGCCAGCGCAGGCGGA AGAAAAAATTTCCCGGGTGAGAAAATTGGCTGAACAGCAAGGACGTGCGATTGAATTCGGCATTCGACTTCATATCA TTGTGCGCGAAACAGAAAAGGAAGCGTGGAATGCTGCGGAACAGCTCATTCGTTATGTCGATGAAAAAACCATTCAG GAAGCACAACGAGTTTTTGCTCGGTATGATTCTGTTGGGCAGCAGCGGATGAGGCAGTTGCATAATGGAAGCAGAGA ATCTCTTGAAATTAGTCCAAATTTATGGGCGGGAGTTGGTCTTGTCAGAGGTGGTGCAGGAACCGCTTTAGTCGGAG ATCCAGAAACAGTAGCTGAGCGGCTATTAGAGTATCATCAATTAGGCATCCGCTACTTTATTTTATCCGGTTACCCG CATTTAGAAGAAGCATACCGGGTAGCTGAACTGCTATTCCCGCTTTTGCCGTTAAACCATAAAAAACAAACAAAACA GTTTATACAAGGGGAAGTTATCGGAAATGAATTTTTTCCTTCATTTATTAAAGTGTGA
orf00957 SEQ ID No.3
TTGGTCATTTCGAAGATTCCGCAAAAAACCGGCTGGTCTTTTGAAGACAATGTTAGATATGCCCAAATT GCTGAAGAAGCCGGATTTGAATATGCGTTGCTGCAGACCAGGTTTGTTGCAAGCTATGGTGCGGAAAATCAACTGGA GGCTATCACGTTGGCGGCAGCTTTAGCGGCGGTAACCAAAAAGCTAAAGCTGATTTCGGCTGTTCTTCCTGGCTTGT GGCATCCGGGGACGGTAGCCAAAATGATCTCTACCATTGACCAAATCAGCCATGGACGTGCGGCGGTTAACATAGTG AGCGGGTGGTTTAAAGGAGAATTCCACGCGTACGGCGAGCCTTGGCTGGATCACGATGAAAGATACCGGAGATCCGA AGAGTTCATCCGTGTGCTGCGCAGCATGTGGACAGAGGAAGAGACCCATTTTCGCGGCGATTTTTACCGAATTAACG GTGCTCCTCTAAAACCGAAGCCATATAACGTTCCGGAGATTTTCCAAGGAGGCAATTCCCGTGCCGCCCGGCAGATG GCTTCCAGAGTGTCCGACTGGTATTTCATGAACGGAAATACGATCGACGGTCTCAAAGCGCAAATCGATGAAGTGTC ATCGCTGGCTAAAGCAAACAACCGCAAGGTGAATTTCGGCGTCAACGCGTTTGTGATCGTACGGGACACGGAAGAGG AAGCGAAGCAGGTGCTTCGCGACATCATTGCATACGCGGATGCCGAAGCCGTGGAAGGCTTCCGCAGCCAAGTGCGC TTTGCCGGCAAAGCCTCCCCAGAAAAAGAAGGCATGTGGGCTCATTCAGATTTTAACGATTTAGTCCAATACAACGA CGGTTTCAAAACCGGATTGATCGGTACGGCGGAACAGGTAGCGGACCGCATTATCGAACTCAAAAAAATCGGCGTCG ATCTCATCCTGACCGGGTTCCTCCATTATGACGAAGACATCCGATTATTCGGCGAAAAAGTGATTCCGTTGGTTCGC GAAAAGGAAGCAGCGTTAAACGTGGATGCCGTATAA
orf00949 SEQ ID No.4
ATGAGATATGGATTTTGGCTGCCGATTTTCGGAGGCTGGCTGCGCAACGTTGAAGATGAAAACATGCCG GCGACGTTTGAATATGCGAAAACGGTGGCGCAAAAAGCGGAACAGTGGGGATATAGCACAACATTAGTCGCAGAGCT GTATTTAAATGATATAAAAGGGCCGGAGAGCGATTCGTTGGAAGCATGGTCAACAGCGGCGGCATTAGCGGCGGTGA CAGAAAAATTGGAAATTATGACGGCAGTCCGCCCTGGTTTCCATAATCCGGCTGTTACGGCAAAAATGGCGGCCAAT ATTGACCACATTAGCAACGGCCGCTTTACGCTGAATATCGTATCAGCATGGTGGGAAGAAGAAGCGCGTCAATATGG CGGTATTTTTACCGAGCATGATAAGCGCTATGATCGCACCGAAGAATTCGTCCAAGTGTTAAAAGGGATGTGGACAA ACGATGTGTTTCATTTCCAAGGGAAATTTTATGAAGTCAAAGGAGCCCATTTAGCGCCAAAGCCGGTGCAAAAGCCG CATCCGATTTTATACGCCGGCGGCGAGAGCGAACGAGGAAAACAAGCGATTGTCGAGCATTGCGATGCGTATGTGAT GCACGGCGGAACAGTCGAAGAAATCGCCCGCAAAGTTGCCGACATGAAACAGCGCCGCCGGCAGGCAGGAAAAGAGC CGTTTCGCTCGTTTGGCATGGCGGCGTTTGTCATTTGCCGCGACAGCGAGGAGGCAGCGCAGGCGGAACTACAGCGT ATTACCGACGTCAAATTATCAAGCGGCTATGCAGGATATCATGATTTTGTCAGCAAATCCCAGCTTGAATTGCAATT AAAATTGCAAGACTATTCTGTATCCAACCGTGGACTGCGGCCAAACTTCGTCGGCACGCCGGAGCAAATTGCCGATC GTATTTTGGCATATGAAAATGCAGGCGTTGATTTATTGTTATTACAATTTTCTCCGCAATTAGAGGAAATGGAGCGG TTTGCCAAACAAGTAATGCCGCTTGTCGAGGAGCGCCGGAACAAGCTCGCGGCAAAAGGGGGAAAGCAAGATGAGCA AAATTTACATTATTCACGAAAATAG
orf01208 SEQ ID No.5
ATGATTTGGCAAACAAGGGTAACGGAACTGCTCGGAATTACATATCCGATTATTCAAGGAGGGCTCGCT TACTTGGCGTACGCTGATTTAGCGGCGGCAGTTTCTAACGCCGGAGGATTGGGACAAATTACAGCCATGTCTCTAGA AACCCCCGGGCAATTGCGCGAAGAAATTCGAAAAGTAAAAGAAAAAACGGATCGTCCATTTGGAGTAAACTTTGCGA TCGGGCAGCATGGCCGCTCTTTTCAGCATATGCTGGAGGCGGCATTGGAAGAAGGGGTACCGGTTGTCTCTGTCACA GGCGGAAATCCTGCTCCGTTTTTCGAACAATTAAAAGGGGTCAACGTGAAAAAACTCGTGCTTGTCGCCGCCGTTCG CCAAGCTGTAAAAGCGGAAGAACTTGGCGCGGACGCCGTCATGGTCGTCGGGCAGGAAGGGGGCGGCCATTTAGGAA AACATGATACTGGGACGTTTGTTCTTATCCCAAAAGTAGTGGACTCCGTTTCCATTCCTGTGATTGCTTCTGGAGGA ATTGGCGACGGGCGCGGGTTGATGGCGGCGTTAGCGCTGGGAGCGGAAGGAGTTGAAATGGGAACGAGATTTATTGC TACAAAAGAGTGCGTCCACGCCCATCCGATTTATAAAGAGATGCTTGTGAACGGTTCTGAACATGATACCGTCGTGA TTAAACGAAGTTTAGGAGCGCCTGGCCGCGCCATCGCCAATGAATGGACGAAAAAAATTTTAGAAATAGAAGAGCAA GGTGGAACGTATGAACAACTAAAAGAATATATCAGCGGAGAAGCGAACCGCCGCTTTATTTATGAAGGAAGAACAAA TGAAGGATTTGCATGGGCGGGGCAAGTGATGGGGCTGATTAAAGACGTACCAACGGTAGCGGAATTGTTTTCGCGAA TGATTAGCGAAGCGGAACAAATTCGGGATCGATGGGCAAACTAA
orf01378 SEQ ID No.6
ATGAATGATGTCTGCCGCTTGTTGCATATTCGTTATCCGATCATTCAAGGGGGAATGGGCAATATTAGT AACGCGCAGCTGGCCAGTGCCGTATCCGAAGCGGGAGGACTTGGAACAATTGGTGTTGGTACAATGCCGCCCGATGA AGTGGAGGAAATCATCATTGAAACAAAACGAAGAACAAGTCAGCCATTTGCCGTTAATATTCCTATTCAGGTAACCC CGTATGTAGACGAAATGATATCGTTAGTCTTAAAACATCGCGTTCCTGTCGTTTCGTTATCAGCGGGGAATCCGGCA CCGCTTATTCCACGTCTTGCGGAACAAGGAGTGAAAATAATTGTCGTGACCGCTTCAGTGAAACAAGCGAAAAAAGC GGAAGCAGCAGGAGCGAATATTATTGTTGCTGAAGGGTATGAAGCTGCGGGAATTAATTCGACGTTAGAGTTGACGA CGATGACGCTGATTCCGCAAATCACCAGCGCTGTCCGTGTTCCCGTCGTAGCTGCCGGCGGAATTGGAGACGGGCGG GGATTGCTTGCCGCGTTTGCACTTGGGGCGCAAGGAGTTCAATTAGGTACGCGCCTCATTGCCACAAAAGATGCGCC GTTTCATGAGACGTACAAACAGTTAATCACCAATGCTAGTGAAAACGAAACAGTGATTGTCGGTCGATCAGTTGGAA GAGTGCGGAGAATTATGCGCACTCCGTATGCAGAGAAATTGCTGCAATATGAAAAGGAAGGAGCGCCGCTTGAAATG TTCAATGAATATACCTCTGAAGATCGTCATCGCCGCGGAGCGCTTCAAGGGGATTTTCATGAAGGATTTGTCAATGC GGGGCAAATTGCCGGACTCATCGAAGACATGCCGACGGTAGCGGAGCTGTTTCGGCAAATGATGGAAGAAGCGAAAC GGCAGCTGCATAAACTACATACACTTTTCCATTCATAA
orf02109 SEQ ID No.7
ATGCATAATTCACATGGATTATGGAAACATCCTGAAAGCAAAAGACAAAGAGGATATAAAGATATTGAT TATTGGATTAAGATGGCAAAACTTTTGGAACGTGGCAAGTTTGATGCCGTTTTTTTCGCGGACGTACTGGGAGTATA TGATACATATCGCCAAAGCAAAGCTCCTTCGATACGTGATGGTCTTCAGTTCCCGGTTAATGATGCAGCATTGATTA TTCCTATTATGGCGAGTGTTACGAAACATTTATCTTTTGCTTTAACAGTTAGTACTACTTATGAACATCCTTTTAGT ACTGCAAGACGTTTTTCCACCCTTGACCATTTGACAAAGGGGCGAATTGCCTGGAATGTCGTCACTTCTTATTTACC AAATGCCGCACGGAACTTCGGGCTTCAAGAAATGATCAAACATGATCAGCGTTACGATATAGCAGACGAATTTCTGG AAGTCGCATATAAACTTTGGGAAGGCAGCTGGGAAGATGATGCATTCATGGAAGATAAACAGAACGGTATATTAATT AATCCAGACAAAGTACATGAAATCAATCATGTCGGAAAATTTTTCTCCGTCGAAGGCCCGCATCTTTGCGAACCATC CCCGCAACGTACACCAGTAATCTATCAAGCAGGCACTTCAGAAAGAGGCAGGGAATTTGCAGCTAAGCATGCTGAGT GTGTTTTCGTTGGAGGGCCTACGCCAGAACGAATTAAATACTATACAAATGACATAAAAAGACGCGCTGAAAAGTAT GGACGTAATCCGGATAATATAAAAGTATTTGCTTTTTTAACGGTTATTGTTGGAAAGACAACGGAAGAAGCAGAACA AAAATTTGCAGAATTAAATCATTTATGGAGCCCAGACGCTTCCAAAGCTCAATTTAGCGGAGCAAGCGGCTATGATC TGGCAGAATATGAAAACAAAGATTTAAATGCCCCTTTTGAATTTAAAAATACAGAGCACGGTCATTATAAAGCGGCT TCTCTTACGAAAGATGCCTCTAAAAGGCTTTCGATAGGCGAAGCGCTTAGAAAGCTTGAACAATTAGACCGAGAGTC TATCATTGTCGGAAATCCAGAAGAAGTGGCAGATGCCATTCAATATCGATTTGAAGCGTCAGGTGTGGATGGATTTA ATCTTAATCATCTGGTTACCCCTTCTAGCCTAGAGGATTTTATTGAATTGGTTATTCCGATACTCCAAAAAAGAGGT TTGTACAAGACGGAATACAAACAAGGAACGTTACGAGAAAAGCTATTTAATCATGGCAGTAGTTTATTACCAGAAGA TCATCCGGGCAGTGCTTATCGCAGTAAATCATTTATTGTGTAA
orf02929 SEQ ID No.8
GTGAGGAAAATGCTGAATACCATTTCTGTTCCAATTATTCAAGCGCCGATGGCTGGAGGTGTTTCAACT CCAGCGCTGGCTGCGGCTGTATCGAATGCAGGAGGGCTTGGTTTTTTGGCAGGTGGTTATAAAACGGCGGAAGAAAT GCGCAAAGAAATTGCCGCAGTCCGGGAGATGACCGATAAGCCGTTTGGCGTTAACGTTTTTGTCCCGAGCGAAGAAG ACGTGGACGAGAAAGAGCTGCTTCGTTATCGACAAGTGTTGGAGAAAGAAGCGGAACGATTCGGCGCGCAGCTTGGG GAGGCAAAATGGGATGATGATGATTGGGAAGCAAAGCTCGCTGTTTTGTATGAAGAAAAAGTTCCTGTAGTAAGTTT TACGTTTGGTTGCCCTTCACCTGATATTATCGCAAAATTGAAAGACAACGGTTCGTTTGTCATCGTGACTGTGACAT CAACGGAAGAAGCGTTGATCGCCAAACAGGCAGGGGCGAACGCATTGTGTGTGCAAGGGTCAGAGGCGGGTGGACAT CGAGCGTCATTTCGCAATAACGCCGCTTCCCATGAAAATGACAGCTTGCTTGTTTTATTGCAACAAATTCGTGAAGC GGCCAATATTCCGCTGGTTGCCGCCGGAGGAATCATGAGCGGGCGCGATATCGCCGCGGTGCTTGCAGCGGGGGCGT GCGCTGCTCAATTAGGTACGGCGTTCTTGCGCTGCCCGGAAAGCGGTGCAAATCCGCTGCATAAAAACGCATTAGTC GATCCGCAATTTTCAGCCACTGCAGTTACCCGCGCCTTTACAGGGCGCCCTGCCCGCGGATTGGTGAACCGCTTCCT GATCGAATATGATAAGCTGGCCCCTGCGGCATATCCGCACATTCACCATATGACAAAGCAGCTTCGCAAAGCAGCTG CTCAAGCAAACGATCCGCAGGCGATGTCTTTATGGGCCGGGCAAGGATATCGCTTGGCGAAAGATATGCCGGCTGGA GAGATTGTACAGTTGTTAATGAAGGAATTGAGAGAGATGGTGAATAAATAG
The present invention is using active somatic cell facture processing high sulfur-containing diesel, slurry oil inferior；It is measured through the bacterium using microcoulomb method The content of sulphur in oil and nitrogen before and after the processing.The result shows that：
（1）Sulphur and nitrogen have remarkable result in W-1 bacterial strains removing oil.
（2）Contained sulfur content is 2.15% in high sulfur-containing diesel without the processing of W-1 bacterial strains, nitrogen content 0.57%, through W-1 bacterium Sulphur, nitrogen content are respectively 0.53%, 0.25% after strain processing.
（3）Contained sulfur content is 1.82% in slurry oil inferior without the processing of W-1 bacterial strains, nitrogen content 0.79%, through W-1 bacterium Sulphur, nitrogen content are respectively 0.72%, 0.32% after strain processing.
Description of the drawings：
Fig. 1 is the colonial morphology figure of W-1.
Specific implementation mode：
Illustrate the present invention with reference to embodiment, the scheme of embodiment described here does not limit the present invention, this field it is special Industry personnel spirit according to the invention can make improvements and change, and the such modifications and variations are regarded as at this In the range of invention, the scope of the present invention and essence are defined by the claims；Wherein agents useful for same is commercially available；
Bacterium source used in the present invention：
Embodiment 1：
（One）The extracting method of hot glucoside ground bacillus W-1
Collection in worksite Dagang Oilfield water sample is added 1% oil using water sample and is enriched under the conditions of 60 DEG C of 180r/min Culture.After a week, in solid LB media（Peptone 10g/L, yeast powder 5g/L, sodium chloride 10g/L, agar powder 15g/L, NaOH tune pH to 7.0,121 DEG C, 20min high pressure sterilizations, when temperature drops to 55 DEG C（Hand is tangible）, pour into culture dish, obtain Obtain solid LB media）Middle dilution spread tablet, after bacterium colony is grown well, growth selection is good, and lawn is larger and morphological feature is each Different bacterium colony, in scribing line separation, purifying on enriched medium solid plate.Continuous several times are crossed after purification, and more plants of pure bacterium are obtained, Pure bacterium verifies its desulfurization removing nitric ability by being seeded in fluid nutrient medium, and obtain one plant has desulfurization removing nitric ability to crude oil The hot glucoside ground bacillus W-1 of bacterial strain（Geobacillus thermoglucosidasius）.
（Two）Microbial name and preservation situation
Hot glucoside ground bacillus W-1（Geobacillus thermoglucosidasius）, in the micro- life of China Object culture presevation administration committee common micro-organisms center（China General Microbiological Cultuer Collection Centre）Preservation is carried out, preserving number is CGMCC 11780.
（Three）The feature of strain and identification.
1, the form and physiological and biochemical property of W-1 bacterial strains
1 mm-25 mm of the colony diameter after 24 h are cultivated on LB culture mediums, milky, bacterium colony is rounded, slightly grand It rises, smooth-shaped, moistening is opaque；Observe under the microscope, cell elongated rod shape, size be 0.3 μm -0.5 μm × 20 μm - 30μm.Gram-positive, peritrichous can move, and gemma end life, round or ellipse is expanded.W-1 cells contact enzyme positive, It can gelatin hydrolysate, starch and casein；NO3-N and NO2-N can be restored；Citrate cannot be utilized；NaC l higher than 3% Growth is suppressed；Optimum growth temperature is 60-65 DEG C, less than 42 DEG C and higher than 75 DEG C.2, W-1 productions acid experiment
Under the conditions of 60 DEG C, with 50 CHB/E carbohydrate indentifying substance items of ABI and its culture medium, by the description of product Book has carried out production acid experiment to W-1 using different carbon source.
As a result show that the bacterial strain fiber can utilize ribose, D- xyloses, L-arabinose, glucose, mannose, fructose, sweet Reveal alcohol, N-Acetyl-D-glucosamine, cellobiose, maltose, sucrose, trehalose, scholar's logical sequence sugar, glycerine, D- cannot be utilized Arabic The fermentation and acids such as sugar, sorbierite, lactose.Identify that this bacterial strain is according to French Mei Liai companies software kitGeobacillus thermoglucosidasius（99.9%, fabulous qualification result）.
3 W-1 of table is to 50 CHB/E sugar culture-medium test results of ABI
Note：+ indicate positive ,-indicating negative, W indicates faint
3, the data for extracting genome sequencing analyze the annotation of 16S rDNA and a series of housekeeping genes, determine thin Bacterium is classified.
The nucleotide sequence of 16S rDNA SEQ ID No.1 as shown in sequence table
ACGAACGCTGGCGGCGTGCCTAATACATGCAAGTCGAGCGGACCGGGCGGGAGCTTGCTTCCGCTTGGTTAGCGGCG GACGGGTGAGTAACACGTGGGTAACCTGCCCGTAAGACCGGGATAACTCCGGGAAACCGGGGCTAATACCGGATAAC ACCGAAGACCGCATGGTCTTCGGTTGAAAGGCGGCTTCGGCTGCCACTTACGGATGGGCCCGCGGCGCATTAGCTAG TTGGTGAGGTAACGGCTCACCAAGGCGACGATGCGTAGCCGGCCTGAGAGGGTGACCGGCCACACTGGGACTGAGAC ACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCGCAATGGACGAAAGTCTGACGGAGCGACGCCGCGT GAGCGAAGAAGGTCTTCGGATCGTAAAGCTCTGTTGTTAGGGAAGAAGAAGTGCCGTTCGAACAGGGCGGCACGGTG ACGGTACCTAACGAGAAAGCCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGGGCGAGCGTTGTCCGG AATTATTGGGCGTAAAGCGCGCGCAGGCGGTCCCTTAAGTC,TGATGTGAAAGCCCACGGCTTAACCGTGGAGGGTC ATTGGAAACTGGGGGACTTGAGTGCAGAAGAGGAGAGCGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGG AGGAACACCAGTGGCGAAGGCGGCTCTCTGGTCTGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAACAGGAT TAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTAGAGGGGTTATTCCCTTTAGTGCTGTAGCTA ACGCGTTAAGCACTCCGCCTGGGGTACGGCCGCAAGGCTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGT GGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCCCTGACAACCCTGGAGACAGG GCGTTCCTCCCTTGCGGGAGGACAGGGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTT AAGTCCCGCAACGAGCGCAACCCTCGCCCCTAGTTGCCAGCATTCAGTTGGGCACTCTAGGGGGACTGCCGGCTAAA AGTCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGG CGGTACAAAGGGCTGCGAACCCGCGAGGGGGAGCGAATCCCAAAAAGCCGCTCTCAGTTCGGATTGCAGGCTGCAAC TCGCCTGCATGAAGCCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACAC ACCGCCCGTCACACCACGAGAGCTTGCAACACCCGAAGTCGGTGAGGTAACCCGCAAGGGAGCCAGCCGCCGAAGGT GGGGCAAGTGATTGGGGTGAAGTCGTAACAAGGTAGCCGTACCGGAAGGTGCGGCTGGATCA
Carry out sequence alignment analysis in GenBank/MBL/DDBJ by the 16S rDNA to W-1, find withGeobacillus thermoglucosidasiusThe similitude highest of BGSC W 95A1 (=ATCC 43742), reaches To 99.22%.
（Four）The application of W-1 bacterial strain correlation desulfurization removing nitric related genes
W-1 bacterial strains are subjected to full genome sequencing, are found from full-length genome and the relevant gene of desulfurization removing nitric, discovery many With desulfurization removing nitric related gene.
In 1 W-1 genomes of table with the relevant gene of desulfurization
In 2 W-1 genomes of table with the relevant gene of denitrogenation
（Five）Desulfurization, denitrification effect
（1）The activation of W-1 strains：W-1 bacterial strains will be deposited in glycerol stocks pipe by 1:1000（v/v）It is seeded to through high pressure In the 50mL culture solutions of sterilizing, 60 DEG C, 150rpm vortex stirrings are closed（Logical oxygen）Culture 18 hours.The group of culture solution therein At and weight percent be：Peptone 1.6%；Yeast extract 1%；Sodium chloride 0.5%；Water surplus.
（2）The expansion culture of W-1 strains：It will（1）The strain of gained activation presses 1:1000（v/v）It is seeded to through high pressure sterilization 50L culture solutions in, 60 DEG C, 150rpm be vortexed stirring, it is closed（Logical oxygen）Culture 18 hours.Culture solution therein is：Inorganic salts Culture medium+final concentration of 1% sucrose solution.The raw material of the minimal medium forms and weight percent is:NH4Cl 0.1%；Na2HPO4·12H2O 0.106%；NaH2PO4·2H2O 0.023%；KCl 0.04%；MgCl·6H2O 0.0092%； FeSO4·7H2O 0.000278%； MnCl2·4H2O 0.000198%；CoSO4·7H2O 0.000281%；CaCl2·2H2O 0.000147%；CuCl2·2H2O 0.000017%；ZnSO4·7H2O 0.000029%；Water surplus.
（3）W-1 strains handle variety classes oil product：
1）Handle high sulfur-containing diesel：It will（2）The 50L bacterium solutions that gained expands after culture are mixed with 20L high sulfur-containing diesels, 60 DEG C, 150rpm, which is vortexed, to be stirred, confined reaction 24 hours；Bacteria-removing liquid, 60 DEG C of pure water, 150rpm is gone to be vortexed stirring clearly after the completion of reaction It is washed till without apparent impurity, collecting that treated, diesel oil is the low nitrogen diesel oil of low-sulfur, is specifically shown in Table 4.
2）Processing slurry oil inferior：It will（2）The 50L bacterium solutions that gained expands after culture are mixed with 25L poor quality slurry oils, 60 DEG C, 150rpm, which is vortexed, to be stirred, confined reaction 24 hours；Bacteria-removing liquid, 60 DEG C of pure water, 150rpm is gone to be vortexed stirring clearly after the completion of reaction It is washed till without apparent impurity, collecting that treated, slurry oil is the low nitrogen slurry oil of low-sulfur.Specifically it is shown in Table 4.
（4）The measurement of present invention total sulfur/total nitrogen content in two kinds of oil product systems：It is examined respectively with WK-2D type microcoulomb instrument Survey two kinds of difference oil products handle without W-1 strains and processed through W-1 strains（3）Total sulfur content, nitrogen pool in gained oil reservoir. Condition：Crack furnace temperature（700 DEG C of stable section, 800 DEG C of burning zone, 600 DEG C of gasification section）, bias ＞ 180mV, amplification factor 200, 2000 Ω of integrating resistor, conversion ratio should be between 75% ~ 115%.
Handling result such as following table：
Table 4：W-1 bacterial strains are to different oil product desulfurizing and denitrifying situation tables
Conclusion：
（1）W-1 bacterial strains are 75.35% to the removal efficiency of the sulphur in high sulfur-containing diesel, and the removal efficiency to the sulphur in slurry oil inferior is 60.34%。
（2）W-1 bacterial strains are 56.14% to the removal efficiency of the nitrogen in high sulfur-containing diesel, and the removal efficiency to the nitrogen in slurry oil inferior is 59.49%。
（3）W-1 bacterial strains have desulfurization removing nitric effect, and significant effect to different oil products.
SEQUENCE LISTING
<110>The Tianjin bio tech ltd Wei Rui
<120>The gene and application of petroleum sweetening denitrogenation bacterial strain and its desulfurization removing nitric
<160> 8
<170> PatentIn version 3.5
<210> 1
<211> 1523
<212> DNA
<213>Artificial sequence
<400> 1
acgaacgctg gcggcgtgcc taatacatgc aagtcgagcg gaccgggcgg gagcttgctt 60
ccgcttggtt agcggcggac gggtgagtaa cacgtgggta acctgcccgt aagaccggga 120
taactccggg aaaccggggc taataccgga taacaccgaa gaccgcatgg tcttcggttg 180
aaaggcggct tcggctgcca cttacggatg ggcccgcggc gcattagcta gttggtgagg 240
taacggctca ccaaggcgac gatgcgtagc cggcctgaga gggtgaccgg ccacactggg 300
actgagacac ggcccagact cctacgggag gcagcagtag ggaatcttcc gcaatggacg 360
aaagtctgac ggagcgacgc cgcgtgagcg aagaaggtct tcggatcgta aagctctgtt 420
gttagggaag aagaagtgcc gttcgaacag ggcggcacgg tgacggtacc taacgagaaa 480
gccccggcta actacgtgcc agcagccgcg gtaatacgta gggggcgagc gttgtccgga 540
attattgggc gtaaagcgcg cgcaggcggt cccttaagtc tgatgtgaaa gcccacggct 600
taaccgtgga gggtcattgg aaactggggg acttgagtgc agaagaggag agcggaattc 660
cacgtgtagc ggtgaaatgc gtagagatgt ggaggaacac cagtggcgaa ggcggctctc 720
tggtctgtaa ctgacgctga ggcgcgaaag cgtggggagc aaacaggatt agataccctg 780
gtagtccacg ccgtaaacga tgagtgctaa gtgttagagg ggttattccc tttagtgctg 840
tagctaacgc gttaagcact ccgcctgggg agtacggccg caaggctgaa actcaaagga 900
attgacgggg gcccgcacaa gcggtggagc atgtggttta attcgaagca acgcgaagaa 960
ccttaccagg tcttgacatc ccctgacaac cctggagaca gggcgttcct cccttgcggg 1020
aggacagggt gacaggtggt gcatggttgt cgtcagctcg tgtcgtgaga tgttgggtta 1080
agtcccgcaa cgagcgcaac cctcgcccct agttgccagc attcagttgg gcactctagg 1140
gggactgccg gctaaaagtc ggaggaaggt ggggatgacg tcaaatcatc atgcccctta 1200
tgacctgggc tacacacgtg ctacaatggg cggtacaaag ggctgcgaac ccgcgagggg 1260
gagcgaatcc caaaaagccg ctctcagttc ggattgcagg ctgcaactcg cctgcatgaa 1320
gccggaatcg ctagtaatcg cggatcagca tgccgcggtg aatacgttcc cgggccttgt 1380
acacaccgcc cgtcacacca cgagagcttg caacacccga agtcggtgag gtaacccgca 1440
agggagccag ccgccgaagg tggggcaagt gattggggtg aagtcgtaac aaggtagccg 1500
taccggaagg tgcggctgga tca 1523
<210> 2
<211> 1128
<212> DNA
<213>Artificial sequence
<400> 2
atggaattat tatggtttat cccttcccat ggtgatgggc ggtatcttgg aacgaccaaa 60
ggaggacgcg ctccggaata cagctatttc cggcaaatcg cccaagcggc tgaccggctt 120
ggctataaag gggtattgat tccgacagga aaatcgtgcg aagatccgtg gctgcttgcg 180
tcagcattag cggcagaaac cgaacaattg cgttttctcg ttgccgtgcg accggggtta 240
atgtctccta cgctagcagc acgcatggct tccacattgg accgcatttc ggaaggccgg 300
ttgcttatca atgtcgtagc tggcggcgat cctgttgagc ttgcgggcga tgggctattt 360
ttaagccatg acgaacgtta tgaagcgacg gatgaatttt taaccgtttg gaaacggctg 420
ttaagcggcg aagaagtcac tttaaaaggg aaacatatcc atgtgaatga ggcaaagctt 480
ctgtttccgc caactcaaaa accgtatccg ccgatttatt ttggcgggtc gtcaccggca 540
ggccaacttg tggcagcaaa acatgcggat gtttacttaa catgggggga accgccagcg 600
caggcggaag aaaaaatttc ccgggtgaga aaattggctg aacagcaagg acgtgcgatt 660
gaattcggca ttcgacttca tatcattgtg cgcgaaacag aaaaggaagc gtggaatgct 720
gcggaacagc tcattcgtta tgtcgatgaa aaaaccattc aggaagcaca acgagttttt 780
gctcggtatg attctgttgg gcagcagcgg atgaggcagt tgcataatgg aagcagagaa 840
tctcttgaaa ttagtccaaa tttatgggcg ggagttggtc ttgtcagagg tggtgcagga 900
accgctttag tcggagatcc agaaacagta gctgagcggc tattagagta tcatcaatta 960
ggcatccgct actttatttt atccggttac ccgcatttag aagaagcata ccgggtagct 1020
gaactgctat tcccgctttt gccgttaaac cataaaaaac aaacaaaaca gtttatacaa 1080
ggggaagtta tcggaaatga attttttcct tcatttatta aagtgtga 1128
<210> 3
<211> 1029
<212> DNA
<213>Artificial sequence
<400> 3
ttggtcattt cgaagattcc gcaaaaaacc ggctggtctt ttgaagacaa tgttagatat 60
gcccaaattg ctgaagaagc cggatttgaa tatgcgttgc tgcagaccag gtttgttgca 120
agctatggtg cggaaaatca actggaggct atcacgttgg cggcagcttt agcggcggta 180
accaaaaagc taaagctgat ttcggctgtt cttcctggct tgtggcatcc ggggacggta 240
gccaaaatga tctctaccat tgaccaaatc agccatggac gtgcggcggt taacatagtg 300
agcgggtggt ttaaaggaga attccacgcg tacggcgagc cttggctgga tcacgatgaa 360
agataccgga gatccgaaga gttcatccgt gtgctgcgca gcatgtggac agaggaagag 420
acccattttc gcggcgattt ttaccgaatt aacggtgctc ctctaaaacc gaagccatat 480
aacgttccgg agattttcca aggaggcaat tcccgtgccg cccggcagat ggcttccaga 540
gtgtccgact ggtatttcat gaacggaaat acgatcgacg gtctcaaagc gcaaatcgat 600
gaagtgtcat cgctggctaa agcaaacaac cgcaaggtga atttcggcgt caacgcgttt 660
gtgatcgtac gggacacgga agaggaagcg aagcaggtgc ttcgcgacat cattgcatac 720
gcggatgccg aagccgtgga aggcttccgc agccaagtgc gctttgccgg caaagcctcc 780
ccagaaaaag aaggcatgtg ggctcattca gattttaacg atttagtcca atacaacgac 840
ggtttcaaaa ccggattgat cggtacggcg gaacaggtag cggaccgcat tatcgaactc 900
aaaaaaatcg gcgtcgatct catcctgacc gggttcctcc attatgacga agacatccga 960
ttattcggcg aaaaagtgat tccgttggtt cgcgaaaagg aagcagcgtt aaacgtggat 1020
gccgtataa 1029
<210> 4
<211> 1095
<212> DNA
<213>Artificial sequence
<400> 4
atgagatatg gattttggct gccgattttc ggaggctggc tgcgcaacgt tgaagatgaa 60
aacatgccgg cgacgtttga atatgcgaaa acggtggcgc aaaaagcgga acagtgggga 120
tatagcacaa cattagtcgc agagctgtat ttaaatgata taaaagggcc ggagagcgat 180
tcgttggaag catggtcaac agcggcggca ttagcggcgg tgacagaaaa attggaaatt 240
atgacggcag tccgccctgg tttccataat ccggctgtta cggcaaaaat ggcggccaat 300
attgaccaca ttagcaacgg ccgctttacg ctgaatatcg tatcagcatg gtgggaagaa 360
gaagcgcgtc aatatggcgg tatttttacc gagcatgata agcgctatga tcgcaccgaa 420
gaattcgtcc aagtgttaaa agggatgtgg acaaacgatg tgtttcattt ccaagggaaa 480
ttttatgaag tcaaaggagc ccatttagcg ccaaagccgg tgcaaaagcc gcatccgatt 540
ttatacgccg gcggcgagag cgaacgagga aaacaagcga ttgtcgagca ttgcgatgcg 600
tatgtgatgc acggcggaac agtcgaagaa atcgcccgca aagttgccga catgaaacag 660
cgccgccggc aggcaggaaa agagccgttt cgctcgtttg gcatggcggc gtttgtcatt 720
tgccgcgaca gcgaggaggc agcgcaggcg gaactacagc gtattaccga cgtcaaatta 780
tcaagcggct atgcaggata tcatgatttt gtcagcaaat cccagcttga attgcaatta 840
aaattgcaag actattctgt atccaaccgt ggactgcggc caaacttcgt cggcacgccg 900
gagcaaattg ccgatcgtat tttggcatat gaaaatgcag gcgttgattt attgttatta 960
caattttctc cgcaattaga ggaaatggag cggtttgcca aacaagtaat gccgcttgtc 1020
gaggagcgcc ggaacaagct cgcggcaaaa gggggaaagc aagatgagca aaatttacat 1080
tattcacgaa aatag 1095
<210> 5
<211> 960
<212> DNA
<213>Artificial sequence
<400> 5
atgatttggc aaacaagggt aacggaactg ctcggaatta catatccgat tattcaagga 60
gggctcgctt acttggcgta cgctgattta gcggcggcag tttctaacgc cggaggattg 120
ggacaaatta cagccatgtc tctagaaacc cccgggcaat tgcgcgaaga aattcgaaaa 180
gtaaaagaaa aaacggatcg tccatttgga gtaaactttg cgatcgggca gcatggccgc 240
tcttttcagc atatgctgga ggcggcattg gaagaagggg taccggttgt ctctgtcaca 300
ggcggaaatc ctgctccgtt tttcgaacaa ttaaaagggg tcaacgtgaa aaaactcgtg 360
cttgtcgccg ccgttcgcca agctgtaaaa gcggaagaac ttggcgcgga cgccgtcatg 420
gtcgtcgggc aggaaggggg cggccattta ggaaaacatg atactgggac gtttgttctt 480
atcccaaaag tagtggactc cgtttccatt cctgtgattg cttctggagg aattggcgac 540
gggcgcgggt tgatggcggc gttagcgctg ggagcggaag gagttgaaat gggaacgaga 600
tttattgcta caaaagagtg cgtccacgcc catccgattt ataaagagat gcttgtgaac 660
ggttctgaac atgataccgt cgtgattaaa cgaagtttag gagcgcctgg ccgcgccatc 720
gccaatgaat ggacgaaaaa aattttagaa atagaagagc aaggtggaac gtatgaacaa 780
ctaaaagaat atatcagcgg agaagcgaac cgccgcttta tttatgaagg aagaacaaat 840
gaaggatttg catgggcggg gcaagtgatg gggctgatta aagacgtacc aacggtagcg 900
gaattgtttt cgcgaatgat tagcgaagcg gaacaaattc gggatcgatg ggcaaactaa 960
<210> 6
<211> 954
<212> DNA
<213>Artificial sequence
<400> 6
atgaatgatg tctgccgctt gttgcatatt cgttatccga tcattcaagg gggaatgggc 60
aatattagta acgcgcagct ggccagtgcc gtatccgaag cgggaggact tggaacaatt 120
ggtgttggta caatgccgcc cgatgaagtg gaggaaatca tcattgaaac aaaacgaaga 180
acaagtcagc catttgccgt taatattcct attcaggtaa ccccgtatgt agacgaaatg 240
atatcgttag tcttaaaaca tcgcgttcct gtcgtttcgt tatcagcggg gaatccggca 300
ccgcttattc cacgtcttgc ggaacaagga gtgaaaataa ttgtcgtgac cgcttcagtg 360
aaacaagcga aaaaagcgga agcagcagga gcgaatatta ttgttgctga agggtatgaa 420
gctgcgggaa ttaattcgac gttagagttg acgacgatga cgctgattcc gcaaatcacc 480
agcgctgtcc gtgttcccgt cgtagctgcc ggcggaattg gagacgggcg gggattgctt 540
gccgcgtttg cacttggggc gcaaggagtt caattaggta cgcgcctcat tgccacaaaa 600
gatgcgccgt ttcatgagac gtacaaacag ttaatcacca atgctagtga aaacgaaaca 660
gtgattgtcg gtcgatcagt tggaagagtg cggagaatta tgcgcactcc gtatgcagag 720
aaattgctgc aatatgaaaa ggaaggagcg ccgcttgaaa tgttcaatga atatacctct 780
gaagatcgtc atcgccgcgg agcgcttcaa ggggattttc atgaaggatt tgtcaatgcg 840
gggcaaattg ccggactcat cgaagacatg ccgacggtag cggagctgtt tcggcaaatg 900
atggaagaag cgaaacggca gctgcataaa ctacatacac ttttccattc ataa 954
<210> 7
<211> 1344
<212> DNA
<213>Artificial sequence
<400> 7
atgcataatt cacatggatt atggaaacat cctgaaagca aaagacaaag aggatataaa 60
gatattgatt attggattaa gatggcaaaa cttttggaac gtggcaagtt tgatgccgtt 120
tttttcgcgg acgtactggg agtatatgat acatatcgcc aaagcaaagc tccttcgata 180
cgtgatggtc ttcagttccc ggttaatgat gcagcattga ttattcctat tatggcgagt 240
gttacgaaac atttatcttt tgctttaaca gttagtacta cttatgaaca tccttttagt 300
actgcaagac gtttttccac ccttgaccat ttgacaaagg ggcgaattgc ctggaatgtc 360
gtcacttctt atttaccaaa tgccgcacgg aacttcgggc ttcaagaaat gatcaaacat 420
gatcagcgtt acgatatagc agacgaattt ctggaagtcg catataaact ttgggaaggc 480
agctgggaag atgatgcatt catggaagat aaacagaacg gtatattaat taatccagac 540
aaagtacatg aaatcaatca tgtcggaaaa tttttctccg tcgaaggccc gcatctttgc 600
gaaccatccc cgcaacgtac accagtaatc tatcaagcag gcacttcaga aagaggcagg 660
gaatttgcag ctaagcatgc tgagtgtgtt ttcgttggag ggcctacgcc agaacgaatt 720
aaatactata caaatgacat aaaaagacgc gctgaaaagt atggacgtaa tccggataat 780
ataaaagtat ttgctttttt aacggttatt gttggaaaga caacggaaga agcagaacaa 840
aaatttgcag aattaaatca tttatggagc ccagacgctt ccaaagctca atttagcgga 900
gcaagcggct atgatctggc agaatatgaa aacaaagatt taaatgcccc ttttgaattt 960
aaaaatacag agcacggtca ttataaagcg gcttctctta cgaaagatgc ctctaaaagg 1020
ctttcgatag gcgaagcgct tagaaagctt gaacaattag accgagagtc tatcattgtc 1080
ggaaatccag aagaagtggc agatgccatt caatatcgat ttgaagcgtc aggtgtggat 1140
ggatttaatc ttaatcatct ggttacccct tctagcctag aggattttat tgaattggtt 1200
attccgatac tccaaaaaag aggtttgtac aagacggaat acaaacaagg aacgttacga 1260
gaaaagctat ttaatcatgg cagtagttta ttaccagaag atcatccggg cagtgcttat 1320
cgcagtaaat catttattgt gtaa 1344
<210> 8
<211> 1044
<212> DNA
<213>Artificial sequence
<400> 8
gtgaggaaaa tgctgaatac catttctgtt ccaattattc aagcgccgat ggctggaggt 60
gtttcaactc cagcgctggc tgcggctgta tcgaatgcag gagggcttgg ttttttggca 120
ggtggttata aaacggcgga agaaatgcgc aaagaaattg ccgcagtccg ggagatgacc 180
gataagccgt ttggcgttaa cgtttttgtc ccgagcgaag aagacgtgga cgagaaagag 240
ctgcttcgtt atcgacaagt gttggagaaa gaagcggaac gattcggcgc gcagcttggg 300
gaggcaaaat gggatgatga tgattgggaa gcaaagctcg ctgttttgta tgaagaaaaa 360
gttcctgtag taagttttac gtttggttgc ccttcacctg atattatcgc aaaattgaaa 420
gacaacggtt cgtttgtcat cgtgactgtg acatcaacgg aagaagcgtt gatcgccaaa 480
caggcagggg cgaacgcatt gtgtgtgcaa gggtcagagg cgggtggaca tcgagcgtca 540
tttcgcaata acgccgcttc ccatgaaaat gacagcttgc ttgttttatt gcaacaaatt 600
cgtgaagcgg ccaatattcc gctggttgcc gccggaggaa tcatgagcgg gcgcgatatc 660
gccgcggtgc ttgcagcggg ggcgtgcgct gctcaattag gtacggcgtt cttgcgctgc 720
ccggaaagcg gtgcaaatcc gctgcataaa aacgcattag tcgatccgca attttcagcc 780
actgcagtta cccgcgcctt tacagggcgc cctgcccgcg gattggtgaa ccgcttcctg 840
atcgaatatg ataagctggc ccctgcggca tatccgcaca ttcaccatat gacaaagcag 900
cttcgcaaag cagctgctca agcaaacgat ccgcaggcga tgtctttatg ggccgggcaa 960
ggatatcgct tggcgaaaga tatgccggct ggagagattg tacagttgtt aatgaaggaa 1020
ttgagagaga tggtgaataa atag 1044

Claims (5)

1. one plant of Geobacillus thermoglucosidasius (Geobacillus thermoglucosidasius), which is characterized in that name Referred to as W-1 has been preserved in " China Committee for Culture Collection of Microorganisms's common micro-organisms center ", preserving number CGMCC 11780。
2. element sulphur side of the Geobacillus thermoglucosidasius described in claim 1 in removing oil organic compounds containing sulfur The application in face.
3. nitrogen side of the Geobacillus thermoglucosidasius described in claim 1 in removing oil organic compounds containing nitrogen The application in face.
4. application of the Geobacillus thermoglucosidasius described in claim 1 in terms of petrochemical industry and environmental protection.
5. the application described in claim 4, petrochemical industry includes oil extraction in oil field, purification containing oil substance；Environmental protection includes Microorganism remediation, fossil fuel deep desulfuration denitrogenation.
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