CN110117559A - One plant of microbacterium W-Y13 and its application - Google Patents
One plant of microbacterium W-Y13 and its application Download PDFInfo
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- CN110117559A CN110117559A CN201910355267.5A CN201910355267A CN110117559A CN 110117559 A CN110117559 A CN 110117559A CN 201910355267 A CN201910355267 A CN 201910355267A CN 110117559 A CN110117559 A CN 110117559A
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- 241001467578 Microbacterium Species 0.000 title claims abstract description 12
- 241000500375 Microbacterium sp. Species 0.000 claims abstract description 44
- 238000004519 manufacturing process Methods 0.000 claims abstract description 15
- 241000894006 Bacteria Species 0.000 claims description 48
- 239000003208 petroleum Substances 0.000 claims description 40
- 239000002068 microbial inoculum Substances 0.000 claims description 34
- 239000000725 suspension Substances 0.000 claims description 31
- 238000011084 recovery Methods 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 14
- 230000015556 catabolic process Effects 0.000 claims description 11
- 238000006731 degradation reaction Methods 0.000 claims description 11
- 238000004945 emulsification Methods 0.000 claims description 7
- 238000002360 preparation method Methods 0.000 claims description 4
- 239000000470 constituent Substances 0.000 claims description 2
- 239000003921 oil Substances 0.000 abstract description 49
- 244000005700 microbiome Species 0.000 abstract description 11
- 239000010779 crude oil Substances 0.000 abstract description 4
- 230000006872 improvement Effects 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 22
- 230000001580 bacterial effect Effects 0.000 description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 16
- 238000000855 fermentation Methods 0.000 description 11
- 230000004151 fermentation Effects 0.000 description 11
- 238000004321 preservation Methods 0.000 description 7
- 108020004465 16S ribosomal RNA Proteins 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 239000001963 growth medium Substances 0.000 description 6
- 238000002347 injection Methods 0.000 description 6
- 239000007924 injection Substances 0.000 description 6
- 230000000813 microbial effect Effects 0.000 description 6
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 6
- 238000006073 displacement reaction Methods 0.000 description 5
- 101150013736 gyrB gene Proteins 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 239000001888 Peptone Substances 0.000 description 4
- 108010080698 Peptones Proteins 0.000 description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 235000019319 peptone Nutrition 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 230000004913 activation Effects 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 230000007483 microbial process Effects 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000009671 shengli Substances 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 238000012549 training Methods 0.000 description 3
- 229910017677 NH4H2 Inorganic materials 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 230000001804 emulsifying effect Effects 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 230000035699 permeability Effects 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241000726221 Gemma Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 241001323976 Microbacterium saccharophilum Species 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 239000006004 Quartz sand Substances 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 235000013681 dietary sucrose Nutrition 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000003129 oil well Substances 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 101150012629 parE gene Proteins 0.000 description 1
- 239000003209 petroleum derivative Substances 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 238000012797 qualification Methods 0.000 description 1
- 238000012113 quantitative test Methods 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000004576 sand Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 229960004793 sucrose Drugs 0.000 description 1
Classifications
-
- 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
- C12N1/20—Bacteria; Culture media therefor
-
- 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
-
- E—FIXED CONSTRUCTIONS
- E21—EARTH OR ROCK DRILLING; MINING
- E21B—EARTH OR ROCK DRILLING; OBTAINING OIL, GAS, WATER, SOLUBLE OR MELTABLE MATERIALS OR A SLURRY OF MINERALS FROM WELLS
- E21B43/00—Methods or apparatus for obtaining oil, gas, water, soluble or meltable materials or a slurry of minerals from wells
- E21B43/16—Enhanced recovery methods for obtaining hydrocarbons
-
- 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
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Geology (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Mining & Mineral Resources (AREA)
- General Health & Medical Sciences (AREA)
- Environmental & Geological Engineering (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Hydrology & Water Resources (AREA)
- Physics & Mathematics (AREA)
- Fluid Mechanics (AREA)
- Water Supply & Treatment (AREA)
- Biodiversity & Conservation Biology (AREA)
- General Life Sciences & Earth Sciences (AREA)
- Geochemistry & Mineralogy (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses one plant of microbacterium W-Y13 and its applications.The classification naming of microbacterium W-Y13 of the invention is microbacterium Microbacterium sp., which is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on January 10th, 2019, and deposit number is CGMCC No.17151.Be experimentally confirmed: microbacterium W-Y13 of the invention can emulsified crude oil, cannot be only used for microbe oil production, improve tar productivity, it may also be used for the improvement and reparation of oil pollution have a good application prospect.
Description
Technical field
The present invention relates to Microbial Enhanced Oil Recovery fields, and in particular to one plant of microbacterium W-Y13 and its application.
Background technique
Oil pollution is widely distributed, and it is various to be related to the various industry and Oil spills etc. such as oil recovery, oil refining, chemical industry, machinery
Pollute environment.Petroleum hydrocarbon complicated components multiplicity, hydrophobicity is strong and is difficult to be contacted by common micro-organisms, degrades, utilizes.It is micro-
Biological oil recovery technique be exactly by injecting nutrition or microorganism into stratum, using the growth metabolism activity of microorganism in oil reservoir,
Improve crude output and recovery ratio.Traditional petroleum function stem separates water body, in soil more.It is existing studies have shown that micro- life
Object removes respectively in water, soil China and foreign countries, also has extensive distribution in petroleum oil phase, therefore separate relevant function from petroleum oil phase
Bacterial strain is of great significance in microbial oil extraction technology.
Summary of the invention
It is an object of the present invention to provide one plant of microbacterium Microbacterium sp.W-Y13.
The deposit number of microbacterium Microbacterium sp.W-Y13 provided by the invention is CGMCC No.17151,
Its classification naming is microbacterium Microbacterium sp., has been preserved in Chinese microorganism strain guarantor on January 10th, 2019
Hide administration committee's common micro-organisms center (abbreviation CGMCC, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Chinese section
Institute of microbiology, institute, postcode 100101).
It is a further object to provide microbacterium Microbacterium sp. or its bacteria suspensions or its culture solution
Or the new application of its tunning or the microbial inoculum containing it.
The present invention provides microbacterium Microbacterium sp. or its bacteria suspensions or its culture solution or its tunning
Or the application containing its microbial inoculum in petroleum recovering.
The present invention also provides microbacterium Microbacterium sp. or its bacteria suspensions or its culture solution or its fermentation to produce
Object or application containing its microbial inoculum in the product for preparing petroleum recovering.
The present invention also provides microbacterium Microbacterium sp. or its bacteria suspensions or its culture solution or its fermentation to produce
Object or application containing its microbial inoculum in microbe oil production.
The present invention also provides microbacterium Microbacterium sp. or its bacteria suspensions or its culture solution or its fermentation to produce
Object or application containing its microbial inoculum in the product for preparing microbe oil production.
The present invention also provides microbacterium Microbacterium sp. or its bacteria suspensions or its culture solution or its fermentation to produce
Object or application containing its microbial inoculum in petroleum emulsification.
The present invention also provides microbacterium Microbacterium sp. or its bacteria suspensions or its culture solution or its fermentation to produce
Object or application containing its microbial inoculum in the product for preparing petroleum emulsification.
The present invention also provides microbacterium Microbacterium sp. or its bacteria suspensions or its culture solution or its fermentation to produce
Object or application containing its microbial inoculum in oil degradation.
The present invention also provides microbacterium Microbacterium sp. or its bacteria suspensions or its culture solution or its fermentation to produce
Object or application containing its microbial inoculum in the product for preparing oil degradation.
The present invention also provides microbacterium Microbacterium sp. or its bacteria suspensions or its culture solution or its fermentation to produce
Object or application containing its microbial inoculum in petroleum pollution.
The present invention also provides microbacterium Microbacterium sp. or its bacteria suspensions or its culture solution or its fermentation to produce
Object or application containing its microbial inoculum in the product for preparing petroleum pollution.
The present invention also provides microbacterium Microbacterium sp. or its bacteria suspensions or its culture solution or its fermentation to produce
Object or containing its microbial inoculum oil pollution reparation in application.
The present invention also provides microbacterium Microbacterium sp. or its bacteria suspensions or its culture solution or its fermentation to produce
Object or containing its microbial inoculum preparation oil pollution reparation product in application.
It is a still further object of the present invention to provide a kind of products.
The active constituent of product provided by the invention is microbacterium Microbacterium sp. or its bacteria suspension or its training
Nutrient solution or its tunning or the microbial inoculum containing it；
The product has following 1) -6) in any function:
1) petroleum recovering；
2) microbe oil production；
3) petroleum emulsification；
4) oil degradation；
5) petroleum pollution；
6) oil pollution reparation.
In above-mentioned application or product, the petroleum can be crude oil；Further, the crude oil can be viscous crude；Further
, the viscous crude can be special thick oil.
It is a still further object of the present invention to provide a kind of methods of microbe oil production.
The method of microbe oil production provided by the invention includes the following steps: during oil recovery, and microbacterium is added
Microbacterium sp. or its bacteria suspension or its culture solution or its tunning or the microbial inoculum containing it.
Final object of the present invention is to provide a kind of method of petroleum pollution and/or reparation.
Petroleum pollution provided by the invention and/or the method for reparation include the following steps: to use microbacterium
Microbacterium sp. or its bacteria suspension or its culture solution or its tunning or containing its microbial inoculum handle oil pollution
Object.
In above-mentioned application or product or method, the microbacterium Microbacterium sp. is microbacterium
Microbacterium sp.W-Y13CGMCC No.17151。
In above-mentioned application or product or method, the microbial inoculum the preparation method is as follows: by microbacterium Microbacterium
Sp.W-Y13CGMCC No.17151 is seeded to LB culture medium, and (it is water that LB, which cultivates based solvent, and solute and its concentration are respectively as follows: 10g/
L NaCl, 10g/L peptone, 5g/L yeast powder, pH 7-8) in culture, obtain the microbial inoculum.Bacteria concentration in the microbial inoculum is not
Lower than 108cfu/mL。
The present invention provides a kind of microbe oil production bacterium W-Y13, classification naming is microbacterium Microbacterium
Sp., which is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on January 10th, 2019,
Deposit number is CGMCC No.17151.Be experimentally confirmed: microbe oil production bacterium W-Y13 of the invention can emulsified crude oil,
It cannot be only used for microbe oil production, improve tar productivity, it may also be used for the improvement and reparation of oil pollution, before there is application well
Scape.
Detailed description of the invention
Fig. 1 is the 16S rDNA Phylogenetic Tree of W-Y13.
Fig. 2 is the gyrB Phylogenetic Tree of W-Y13.
Fig. 3 is that W-Y13 emulsifies special thick oil.A left side is W-Y13, and the right side is control.
Preservation explanation
Latin name: microbacterium Microbacterium sp.
Strain number: W-Y13
Preservation mechanism: China Committee for Culture Collection of Microorganisms's common micro-organisms center
Preservation mechanism abbreviation: CGMCC
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date: on January 10th, 2019
Collection is registered on the books number: CGMCC No.17151
Specific embodiment
Embodiment below facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments
Method is unless otherwise specified conventional method.Test material as used in the following examples is unless otherwise specified certainly
What routine biochemistry reagent shop was commercially available.Quantitative test in following embodiment is respectively provided with three repeated experiments, as a result makes even
Mean value.
LB culture medium prescription in following embodiments is as follows: solvent is water, and solute and its concentration are respectively as follows: 10g/L
NaCl, 10g/L peptone, 5g/L yeast powder, pH 7-8.
The deposit number of CGMCC No.14243 bacterial strain in following embodiments is CGMCC No.14243, and classification naming is
Microbacterium saccharophilum was preserved in Chinese microorganism strain preservation management on 06 14th, 2017
(abbreviation CGMCC, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, the Chinese Academy of Sciences are micro- for committee's common micro-organisms center
Biological study institute, postcode 100101).CGMCC No.14243 bacterial strain is recorded in a kind of entitled " oil reservoir origin microbacterium
And its application in oil exploitation ", application No. is 201710759313.9, publication No. be 107523517 A of CN patent text
In offering.
Embodiment 1, the separation of W-Y13 bacterial strain, identification and preservation
One, the separation of W-Y13 bacterial strain
In September, 2017, obtains oily phase petroleum from the oil well produced liquid of the Shengli Oil Field of Karamay, Xinjiang, inventor from
In isolated one plant of bacterium, be named as bacterial strain W-Y13.
Two, the identification of W-Y13 bacterial strain
1, the Morphological Identification of W-Y13 bacterial strain
The morphological feature of W-Y13 bacterial strain: being wet raised yellow color colonies on LB solid plate, and thallus is that rod-short does not produce
Gemma.
2, the Physiology and biochemistry identification of W-Y13 bacterial strain
The physiological and biochemical property of W-Y13 bacterial strain: 30-35 DEG C of temperature can be suitble to using dextrose and saccharose as carbon source.
3, the Molecular Identification of W-Y13 bacterial strain
By the W-Y13 strain inoculated of activation in LB culture medium, 30 DEG C, 150rpm shaken cultivation for 24 hours.Take the fresh training of 1mL
4 DEG C of bacteria liquid, 8000rpm are centrifuged 5min, collect thallus in 2mL centrifuge tube, extract DNA using DNA extraction kit.Electrophoresis
After detection, 16S rDNA gene universal primer 8F/1492R and gyrB gene universal primer up1F/up2R is respectively adopted and carries out PCR
Amplification.After PCR product electrophoresis detection, 16S rDNA gene order and gyrB gene order, the specific sequence of 16S rDNA gene are measured
Column are as shown in sequence 1 in sequence table, and gyrB gene particular sequence is as shown in sequence 2 in sequence table.
16S rDNA gene order belongs to Microbacterium bacterium after comparing, and is used based on 16S rDNA gene soft
The phylogenetic tree of part MEGA building is as shown in Figure 1.The phylogenetic tree such as Fig. 2 constructed based on gyrB gene using software MEGA
It is shown.
In summary qualification result shows that bacterial strain W-Y13 is a kind of newfound Microbacterium bacterium.
Three, the preservation of W-Y13 bacterial strain
The classification naming of W-Y13 bacterial strain is microbacterium Microbacterium sp., and the bacterial strain is on January 10th, 2019
It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (abbreviation CGMCC, address: Chaoyang District, Beijing City
The institute 3 of North Star West Road 1, Institute of Microorganism, Academia Sinica, postcode 100101), deposit number is CGMCC No.17151.
Embodiment 2, W-Y13 bacterial strain emulsify special thick oil
Shengli Oil Field of the petroleum in the acquisition of in September, 2017 from Xinjiang carat Ma in the present embodiment, 30 DEG C of oil viscosity
For 35076mPa.s, belong to special thick oil.
1, the W-Y13 after activation is seeded in LB culture medium, 30 DEG C, 150rpm shaken cultivation for 24 hours, obtain W-Y13 bacterium
Liquid (bacteria concentration >=108cfu/mL)。
2, test group: by 5 parts by volume bacterium solutions be seeded to 95 parts by volume petroleum-minimal medium (solvent is water, solute and
Its concentration is respectively 1g/L petroleum, 5g/L NaCl, 1g/L NH4H2PO4、1g/L(NH4)2SO4、1g/L K2HPO4、3g/L
KNO3, pH 7-8) in, 30 DEG C, 150rpm shaken cultivation 15d.
Control group 1: by 100 parts by volume petroleum-minimal medium, 30 DEG C, 150rpm shaken cultivation 15d.
Control group 2: being seeded in 95 parts by volume petroleum-minimal medium for the CGMCC No.14243 bacterium solution of 5 parts by volume,
30 DEG C, 150rpm shaken cultivation 15d.Wherein, CGMCC No.14243 bacterium solution is the CGMCC No.14243 inoculation after activating
Into LB culture medium, 30 DEG C, bacterium solution (bacteria concentration >=10 that obtain afterwards for 24 hours of 150rpm shaken cultivation8cfu/mL)。
3, after completing step 2, photographic result is shown in Fig. 3.In Fig. 3, a left side 1 is test group, and a left side 2 is control group 1, a left side 3 is control
Group 2.There is apparent emulsification to special thick oil at 30 DEG C in bacterial strain W-Y13；And CGMCC No.14243 does not have emulsifying effectiveness,
Speculate that CGMCC No.14243 only has emulsifying effectiveness to thin oil (viscosity is less than 50mPa.s).
4, it is completed in the system of step 2 according to the method measurement in document " separation of oil degradation bacterium and its degradation property "
Petroleum residual amount, calculate petroleum degradation rate.Petroleum degradation rate=(petroleum additional amount-petroleum residual amount)/petroleum additional amount.Meter
Calculate the result shows that: petroleum degradation rate 41.8%.
Embodiment 3, the application of W-Y13 bacterial strain in microbial oil extraction technology
Shengli Oil Field of the petroleum in the acquisition of in September, 2017 from Xinjiang carat Ma in the present embodiment, 30 DEG C of oil viscosity
For 35076mPa.s, belong to special thick oil.
The present embodiment passes through the oil displacement efficiency that indoor object mould oil displacement experiment evaluates microorganism fungus kind.Specific experiment method is such as
Under:
1, using High Pressure Model pipe (specificationPurchased from Hai'an petroleum scientific research Instrument Ltd.) in load
The quartz sand of different meshes is made as sandpack column pipe.Using permeability measuring apparatus measurement model pipe permeability, selection meets
Model pipe.Saturation water flooding is vacuumized, porosity is calculated；The parameter of the model pipe for subsequent step finally prepared is shown in Table
1。
Table 1, model pipe back-up sand parameter
2, petroleum is persistently injected from model tube inlet, until model pipe outlet detects that petroleum flows out, measures initial oil
Saturation degree；And aging 3 days under reservoir temperature (30 DEG C).
3, the displacement of reservoir oil is carried out with injection water, moisture content (98%) stops water drive until outlet production fluid reaches capacity, and calculates harvesting
Rate (as waterflood recovery efficiency factor).
4, according to the injection rate injection infected water of 1 model pipe pore volume volume (after completing injection, calculate recovery ratio,
As infuse microbial process recovery ratio), it is cultivated 30 days under reservoir temperature (30 DEG C).
Infected water the preparation method is as follows: the W-Y13 of activation is seeded in LB culture medium, 30 DEG C, 150rpm oscillation training
It supports for 24 hours, obtains W-Y13 bacterium solution (bacteria concentration >=108cfu/mL).W-Y13 bacterium solution, NH is added in injection water4H2PO4、(NH4)2SO4、K2HPO4、KNO3, peptone and yeast powder, make it in the middle concentration of infected water be respectively 5% (volume fraction) W-Y13 bacterium
Liquid, 1g/L NH4H2PO4、1g/L(NH4)2SO4、1g/L K2HPO4、3g/L KNO3, 2g/L peptone and 1g/L yeast powder.
5, the displacement of reservoir oil is carried out with injection water again, moisture content (98%) stops water drive until outlet production fluid reaches capacity, and calculating is adopted
Yield (as microbial action subsequent waterflooding recovery ratio).
To in water drive, note microbial process, the recovery ratio of the different phases such as microbial action subsequent waterflooding compared,
It the results are shown in Table 2.The indoor object mould displacement of reservoir oil shows that recovery ratio 6.11% can be improved using W-Y13 bacterial strain.
Table 2, recovery ratio
| Object mode step section | Recovery ratio (%) | It improves recovery ratio (%) |
| Water drive | 36.51 | / |
| Infuse microbial process | 38.64 | 2.13 |
| Microbial action subsequent waterflooding | 42.62 | 6.11 |
Sequence table
<110>Beijing Run Shi Enertech Co., Ltd.
<120>one plants of microbacterium W-Y13 and its applications
<160>2
<170>PatentIn version 3.5
<210>1
<211>1379
<212>DNA
<213>artificial sequence (Artificial Sequence)
<400>1
tgcagtcgaa cggtgaacac ggagcttgct ctgtgggatc agtggcgaac gggtgagtaa 60
cacgtgagca acctgcccct gactctggga taagcgctgg aaacggcgtc taatactgga 120
tatgtgacgt gatcgcatgg tctgcgtctg gaaagaattt cggttgggga tgggctcgcg 180
gcctatcagc ttgttggtga ggtaatggct caccaaggcg tcgacgggta gccggcctga 240
gagggtgacc ggccacactg ggactgagac acggcccaga ctcctacggg aggcagcagt 300
ggggaatatt gcacaatggg cgcaagcctg atgcagcaac gccgcgtgag ggatgacggc 360
cttcgggttg taaacctctt ttagcaggga agaagcgaaa gtgacggtac ctgcagaaaa 420
agcgccggct aactacgtgc cagcagccgc ggtaatacgt agggcgcaag cgttatccgg 480
aattattggg cgtaaagagc tcgtaggcgg tttgtcgcgt ctgctgtgaa atccggaggc 540
tcaacctccg gcctgcagtg ggtacgggca gactagagtg cggtagggga gattggaatt 600
cctggtgtag cggtggaatg cgcagatatc aggaggaaca ccgatggcga aggcagatct 660
ctgggccgta actgacgctg aggagcgaaa gggtggggag caaacaggct tagataccct 720
ggtagtccac cccgtaaacg ttgggaacta gttgtggggt ccattccacg gattccgtga 780
cgcagctaac gcattaagtt ccccgcctgg ggagtacggc cgcaaggcta aaactcaaag 840
gaattgacgg ggacccgcac aagcggcgga gcatgcggat taattcgatg caacgcgaag 900
aaccttacca aggcttgaca tatacgagaa cgggccagaa atggtcaact ctttggacac 960
tcgtaaacag gtggtgcatg gttgtcgtca gctcgtgtcg tgagatgttg ggttaagtcc 1020
cgcaacgagc gcaaccctcg ttctatgttg ccagcacgta atggtgggaa ctcatgggat 1080
actgccgggg tcaactcgga ggaaggtggg gatgacgtca aatcatcatg ccccttatgt 1140
cttgggcttc acgcatgcta caatggccgg tacaaagggc tgcaataccg cgaggtggag 1200
cgaatcccaa aaagccggtc ccagttcgga ttgaggtctg caactcgacc tcatgaagtc 1260
ggagtcgcta gtaatcgcag atcagcaacg ctgcggtgaa tacgttcccg ggtcttgtac 1320
acaccgcccg tcaagtcatg aaagtcggta acacctgaag ccggtggcct aacccttgt 1379
<210>2
<211>888
<212>DNA
<213>artificial sequence (Artificial Sequence)
<400>2
acaaggggct gcgcatcgag ctgaaggacg agcgggagtc ctcggcctac gaggtggagg 60
aggacggcgc gaccgtcacc cgccagccga gcgacgtgtt cttctacgag cgcggcctcg 120
tcgactacgt cgagtacctc aacaaggtcc gccatgccga ggtcgtgaac gacgagatca 180
tcgcgttcga gtccgaggac accgagcgga agatctcgct cgaggtcgcg atgcagtgga 240
ccacctcgta caccgagaac gtgttcacct acgccaacac gatcaacacg cacgagggag 300
gcacccacga ggagggcttc cgcgcggcgc tgaccactct cgtgaacaag tacgcccgcg 360
cgaacaacct gctcaaggag aaggacgaca acctctccgg cgacgacgtg cgcgagggac 420
tgacggccgt catctcgatc aagctcggcg agccccagtt cgaggggcag accaagacga 480
agctcggcaa caccgaggcg aaggcgttcg tgcagaaggt cgtcggcgat cagctcggcg 540
actggttcga tcgcaacccg aaccaggcga agaacatcat ccgcaaggcg atcgacgcgg 600
ccaccgcccg tctcgccgcg cgcaaggccc gggagaccgc gcggcgcaag agcgtcttcg 660
agtcggcggc catgccggac aagctcaagg actgcacgag caaggatccg tcgatcagcg 720
agatcttcct ggtggagggt gactccgccg gcggttcggc cgtgcagggg cgcgacccgc 780
acacccaggc gatcctcgcg ctgcgcggca agatcctcaa cgtggagcgt gcacgtctcg 840
acaaggcttt gggtaacaag gaagtccagg cgatgatcca ggccttcg 888
Claims (10)
1. one plant of microbacterium Microbacterium sp.W-Y13, deposit number is CGMCC No.17151.
2. microbacterium Microbacterium sp. or its bacteria suspension or its culture solution or its tunning or the microbial inoculum containing it
Application in petroleum recovering；
Or, microbacterium Microbacterium sp. or its bacteria suspension or its culture solution or its tunning or the microbial inoculum containing it
Application in the product for preparing petroleum recovering.
3. microbacterium Microbacterium sp. or its bacteria suspension or its culture solution or its tunning or the microbial inoculum containing it
Application in microbe oil production；
Or, microbacterium Microbacterium sp. or its bacteria suspension or its culture solution or its tunning or the microbial inoculum containing it
Application in the product for preparing microbe oil production.
4. microbacterium Microbacterium sp. or its bacteria suspension or its culture solution or its tunning or the microbial inoculum containing it
Application in petroleum emulsification；
Or, microbacterium Microbacterium sp. or its bacteria suspension or its culture solution or its tunning or the microbial inoculum containing it
Application in the product for preparing petroleum emulsification.
5. microbacterium Microbacterium sp. or its bacteria suspension or its culture solution or its tunning or the microbial inoculum containing it
Application in oil degradation；
Or, microbacterium Microbacterium sp. or its bacteria suspension or its culture solution or its tunning or the microbial inoculum containing it
Application in the product for preparing oil degradation.
6. microbacterium Microbacterium sp. or its bacteria suspension or its culture solution or its tunning or the microbial inoculum containing it
Application in petroleum pollution；
Or, microbacterium Microbacterium sp. or its bacteria suspension or its culture solution or its tunning or the microbial inoculum containing it
Application in the product for preparing petroleum pollution.
7. microbacterium Microbacterium sp. or its bacteria suspension or its culture solution or its tunning or the microbial inoculum containing it
Application in oil pollution reparation；
Or, microbacterium Microbacterium sp. or its bacteria suspension or its culture solution or its tunning or the microbial inoculum containing it
Application in the product of preparation oil pollution reparation.
8. a kind of product, active constituent is microbacterium Microbacterium sp. or its bacteria suspension or its culture solution or its hair
Ferment product or microbial inoculum containing it；
The product has following 1) -6) in any function:
1) petroleum recovering；
2) microbe oil production；
3) petroleum emulsification；
4) oil degradation；
5) petroleum pollution；
6) oil pollution reparation.
9. a kind of method of microbe oil production, includes the following steps: during oil recovery, microbacterium is added
Microbacterium sp. or its bacteria suspension or its culture solution or its tunning or the microbial inoculum containing it；
Or, a kind of petroleum pollution and/or the method for reparation, include the following steps: with microbacterium Microbacterium
Sp. or its bacteria suspension or its culture solution or its tunning or petroleum pollution is handled containing its microbial inoculum.
10. according to any application of claim 2-7 or product according to any one of claims 8 or side as claimed in claim 9
Method, it is characterised in that: the microbacterium Microbacterium sp. is microbacterium Microbacterium sp.W-Y13
CGMCC No.17151。
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| CN201910355267.5A CN110117559B (en) | 2019-04-29 | 2019-04-29 | Microbacterium W-Y13 and application thereof |
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| CN201910355267.5A CN110117559B (en) | 2019-04-29 | 2019-04-29 | Microbacterium W-Y13 and application thereof |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104342392A (en) * | 2014-10-30 | 2015-02-11 | 中国石油天然气股份有限公司 | Microbacterium oxydans for degrading polycyclic aromatic hydrocarbon and application thereof |
| CN104651280A (en) * | 2015-03-02 | 2015-05-27 | 南京大学 | Petroleum degrading strain P-6 and application thereof |
| CN107523517A (en) * | 2017-08-30 | 2017-12-29 | 中国石油天然气股份有限公司 | A kind of oil reservoir origin microbacterium and its application in oil exploitation |
| CN108046441A (en) * | 2017-10-30 | 2018-05-18 | 大连民族大学 | The degraded oil of bacterial strain P51 and the method for phenanthrene |
-
2019
- 2019-04-29 CN CN201910355267.5A patent/CN110117559B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104342392A (en) * | 2014-10-30 | 2015-02-11 | 中国石油天然气股份有限公司 | Microbacterium oxydans for degrading polycyclic aromatic hydrocarbon and application thereof |
| CN104651280A (en) * | 2015-03-02 | 2015-05-27 | 南京大学 | Petroleum degrading strain P-6 and application thereof |
| CN107523517A (en) * | 2017-08-30 | 2017-12-29 | 中国石油天然气股份有限公司 | A kind of oil reservoir origin microbacterium and its application in oil exploitation |
| CN108046441A (en) * | 2017-10-30 | 2018-05-18 | 大连民族大学 | The degraded oil of bacterial strain P51 and the method for phenanthrene |
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| CN110117559B (en) | 2022-12-02 |
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