CN110079479B - Tenebriomonas lutescens W-Y14 and application thereof - Google Patents
Tenebriomonas lutescens W-Y14 and application thereof Download PDFInfo
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Abstract
The invention discloses a xanthomonas tenosynoviae W-Y14 and application thereof. The Xanthomonas rattan W-Y14 is classified and named as Xanthomonas rattan Luteimonas sp, and the strain is preserved in the China general microbiological culture Collection center in 2019, 1 month and 10 days, and the preservation number is CGMCC No. 17152. Experiments prove that: the xanthomonas tenore W-Y14 can emulsify crude oil, can be used for microbial oil recovery and oil recovery rate improvement, can also be used for treatment and restoration of petroleum pollution, and has good application prospect.
Description
Technical Field
The invention relates to the technical field of microbial oil recovery, in particular to a luteolomonas lutea W-Y14 and application thereof.
Background
The petroleum pollution is widely distributed, and relates to various industries such as oil extraction, oil refining, chemical engineering, machinery and the like, and various polluted environments such as petroleum leakage and the like. The petroleum hydrocarbon components are complex and diverse, and have strong hydrophobicity, so that the petroleum hydrocarbon components are difficult to be contacted, degraded and utilized by common microorganisms. The microbial oil recovery technology is to inject nutrients or microbes into the stratum and to utilize the growth and metabolism of microbes in oil deposit to raise the yield and recovery rate of crude oil. The traditional petroleum functional bacterial strain is separated from water and soil. The existing research shows that the microorganisms are widely distributed in the petroleum oil phase besides water and soil respectively, so that the separation of related functional strains from the petroleum oil phase has important significance in the microbial oil recovery technology.
Disclosure of Invention
One purpose of the invention is to provide a strain of Xanthomonas rattus Luteimonas sp.W-Y14.
The collection number of the xanthomonas luteoimmunas sp.W-Y14 provided by the invention is CGMCC No.17152, which is classified and named as the xanthomonas luteoimuns sp.and is preserved in the common microorganism center of China Committee for culture Collection of microorganisms (CGMCC for short, the address: No. 3 of West Siro 1 of the sunward area of Beijing, institute of microbiology of China academy of sciences, postal code 100101) in 1 month and 10 days of 2019.
Another purpose of the invention is to provide a new use of the luceimonas sp.
The invention provides application of Xanthomonas rattan Luteimonas sp or bacterial suspension thereof or culture solution thereof or fermentation product thereof or microbial inoculum containing the same in petroleum recovery.
The invention also provides application of the xanthomonas tenore Luteimonas sp or bacterial suspension thereof or culture solution thereof or fermentation product thereof or microbial inoculum containing the same in preparation of petroleum products.
The invention also provides application of the xanthomonas tenore Luteimonas sp or bacterial suspension thereof or culture solution thereof or fermentation product thereof or microbial inoculum containing the same in microbial oil recovery.
The invention also provides application of the xanthomonas tenore Luteimonas sp or bacterial suspension thereof or culture solution thereof or fermentation product thereof or microbial inoculum containing the same in preparation of microbial oil recovery products.
The invention also provides application of the xanthomonas tenore Luteimonas sp or bacterial suspension thereof or culture solution thereof or fermentation product thereof or microbial inoculum containing the same in petroleum emulsification.
The invention also provides application of the xanthomonas tenore Luteimonas sp or bacterial suspension thereof or culture solution thereof or fermentation product thereof or microbial inoculum containing the same in preparation of petroleum emulsified products.
The invention also provides application of the xanthomonas tenore Luteimonas sp or bacterial suspension thereof or culture solution thereof or fermentation product thereof or microbial inoculum containing the same in petroleum degradation.
The invention also provides application of the xanthomonas tenore Luteimonas sp or bacterial suspension thereof or culture solution thereof or fermentation product thereof or microbial inoculum containing the same in preparation of petroleum degradation products.
The invention also provides application of the xanthomonas tenore Luteimonas sp or bacterial suspension thereof or culture solution thereof or fermentation product thereof or microbial inoculum containing the same in petroleum pollution treatment.
The invention also provides application of the xanthomonas tenore Luteimonas sp or bacterial suspension thereof or culture solution thereof or fermentation product thereof or microbial inoculum containing the same in preparing products for treating petroleum pollution.
The invention also provides application of the xanthomonas tenore Luteimonas sp or bacterial suspension thereof or culture solution thereof or fermentation product thereof or microbial inoculum containing the same in petroleum pollution remediation.
The invention also provides application of the xanthomonas tenore Luteimonas sp or bacterial suspension thereof or culture solution thereof or fermentation product thereof or microbial inoculum containing the same in preparation of products for repairing petroleum pollution.
It is a further object of the invention to provide a product.
The active component of the product provided by the invention is Xanthomonas rattan Luteimonas sp or the bacterial suspension thereof or the culture solution thereof or the fermentation product thereof or the microbial inoculum containing the same;
the product has any one of the following functions 1) to 6):
1) oil recovery;
2) microbial oil recovery;
3) petroleum emulsification;
4) degrading petroleum;
5) treating petroleum pollution;
6) and (4) repairing petroleum pollution.
In the above application or product, the petroleum may be crude oil; further, the crude oil may be a thick oil.
It is yet another object of the present invention to provide a method for microbial oil recovery.
The microbial oil recovery method provided by the invention comprises the following steps: in the oil extraction process, the Luteimonas sp, or the bacterial suspension thereof, or the culture solution thereof, or the fermentation product thereof, or the microbial inoculum containing the Luteimonas sp is added.
It is a final object of the present invention to provide a method for remediation and/or rehabilitation of petroleum pollution.
The method for treating and/or repairing petroleum pollution provided by the invention comprises the following steps: the petroleum pollutants are treated by using the xanthomonas tenofovir sp, or a bacterial suspension thereof, or a culture solution thereof, or a fermentation product thereof, or a microbial inoculum containing the same.
In the above application or product or method, the luceimonas sp. of Tenacia is Luteimonas sp.W-Y14 CGMCC No. 17152.
In the above application or product or method, the preparation method of the microbial inoculum is as follows: the strain is obtained by inoculating the Xanthomonas rattus Luteimonas sp.W-Y14 CGMCC No.17152 into an LB culture medium (the solvent of the LB culture medium is water, the solute and the concentration thereof are respectively 10g/L NaCl, 10g/L peptone and 5g/L yeast powder, and the pH value is 7-8) for culture. The concentration of the bacteria in the microbial inoculum is not less than 108cfu/mL。
The invention provides a microbial oil recovery bacterium W-Y14, which is classified and named as Xanthomonas luteinii sp, and the strain is preserved in the China general microbiological culture Collection center in 2019, 1 month and 10 days, and the preservation number is CGMCC No. 17152. Experiments prove that: the microbial oil recovery bacterium W-Y14 can emulsify crude oil, can be used for microbial oil recovery and oil recovery rate improvement, can be used for treatment and restoration of oil pollution, and has good application prospect.
Drawings
FIG. 1 shows a phylogenetic tree of the 16S rDNA gene of W-Y14.
FIG. 2 shows W-Y14 emulsified thick oil. The left is W-Y14 and the right is control.
Deposit description
Latin name: xanthomonas rattus Luteimonas sp.
The strain number is as follows: W-Y14
The preservation organization: china general microbiological culture Collection center
The preservation organization is abbreviated as: CGMCC (China general microbiological culture Collection center)
Address: xilu No.1 Hospital No. 3 of Beijing market facing Yang district
The preservation date is as follows: 1 month and 10 days 2019
Registration number of the preservation center: CGMCC No.17152
Detailed Description
The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified. The quantitative tests in the following examples, all set up three replicates and the results averaged.
The formulation of LB medium in the following examples is as follows: the solvent is water, and the solute and the concentration thereof are respectively: 10g/L NaCl, 10g/L peptone and 5g/L yeast powder, and the pH value is 7-8.
Example 1 isolation, identification and preservation of the W-Y14 Strain
Isolation of the first, W-Y14 Strain
In 9 months in 2017, oil phase petroleum is obtained from oil well produced liquid of Xinjiang oil field of Xinjiang Clamayi, and the inventor separates a strain from the oil phase petroleum and named the strain W-Y14.
Identification of the two, W-Y14 Strain
1. Morphological characterization of W-Y14 Strain
Morphological characteristics of the W-Y14 strain: irregular, raised, bright yellow colonies formed on the LB solid plates.
2. Physiological and biochemical identification of W-Y14 Strain
Physiological and biochemical characteristics of the W-Y14 strain: growth was carried out with 0-5% sodium chloride, and glucose and galactose were able to be utilized as carbon sources.
3. Molecular characterization of the W-Y14 Strain
The activated W-Y14 strain was inoculated in LB medium and cultured with shaking at 150rpm at 30 ℃ for 24 hours. 1mL of fresh culture solution is taken, centrifuged for 5min at 4 ℃ and 8000rpm, the thalli are collected in a 2mL centrifuge tube, and DNA is extracted by adopting a DNA extraction kit. After electrophoretic detection, 16S rDNA gene universal primer 8F/1492R is adopted for PCR amplification. And (3) after electrophoresis detection of the PCR product, determining a 16S rDNA gene sequence, wherein the specific sequence of the 16S rDNA gene is shown as a sequence 1 in a sequence table.
The 16S rDNA gene sequence belongs to Luteimonas after alignment, but has lower similarity with the existing strain. The phylogenetic tree was constructed using the software MEGA as shown in figure 1. The comparison of the physiological and biochemical characteristics of W-Y14 and the strain Luteimonas compositi with the highest similarity is shown in Table 1, wherein, -means no growth and + means growth.
Table 1, W-Y14 comparison of physiological and biochemical characteristics of the strain Luteimonas composti with highest similarity
| Numbering | 6% sodium chloride | Sodium acetate |
| W-Y14 | - | - |
| Luteimonas composti | + | + |
The results of the above identification are combined to show that the strain W-Y14 is a newly discovered Luteimonas.
Deposit of the three, W-Y14 Strain
The W-Y14 strain is classified and named as Xanthomonas luteinis sp, and the strain is preserved in China general microbiological culture Collection center (CGMCC for short, the address: Beijing university Hokko No.1, West Lu No. 3, China academy of sciences microbial research institute, postal code 100101) in 1 month and 10 days in 2019, and the preservation number is CGMCC No. 17152.
Example 2 emulsification of crude oil by W-Y14 Strain
The oil in this example was collected from Xinjiang oil fields of Xinjiang kramey in 2017, 9 months, and the oil viscosity at 50 ℃ was 191.9 mPa.s.
1. Inoculating activated W-Y14 into LB culture mediumPerforming shaking culture at 30 deg.C and 150rpm for 24 hr to obtain W-Y14 bacterial solution (bacterial concentration is not less than 10)8cfu/mL)。
2. Test groups: inoculating 5 volume parts of the bacterial liquid to 95 volume parts of petroleum-inorganic salt culture medium (the solvent is water, the solute and the concentration thereof are respectively 1g/L petroleum, 5g/L NaCl and 1g/L NH)4H2PO4、1g/L(NH4)2SO4、1g/L K2HPO4、 3g/L KNO3pH 7-8), 30 ℃ and 150rpm for 7 d.
Control group: 100 parts by volume of the petroleum-inorganic salt medium was subjected to shaking culture at 30 ℃ and 150rpm for 7 days.
3. After completion of step 2, the photographic results are shown in FIG. 2. In FIG. 2, the left side shows the test group, and the right side shows the control group. The strain W-Y14 has obvious emulsification effect on the thick oil at 30 ℃.
4. And (3) measuring the residual petroleum amount in the system for completing the step (2) according to a method in the literature 'separation of crude oil degrading bacteria and degradation performance thereof', and calculating the petroleum degradation rate. Petroleum degradation rate (petroleum addition-petroleum residue)/petroleum addition. The calculation result shows that: the petroleum degradation rate is 48.4%.
Example 3 application of the W-Y14 Strain in microbial oil recovery technology
The oil in this example was collected from Xinjiang oil fields of Xinjiang kramey in 2017, 9 months, and the oil viscosity at 50 ℃ was 191.9 mPa.s.
In the embodiment, the oil displacement effect of the microbial strains is evaluated through an indoor physical model oil displacement experiment. The specific experimental method is as follows:
1. using high pressure die-type pipe (specification)
Purchased from haian oil research instruments ltd) filled with quartz sand of different mesh numbers to form sand-filled mold tubes. And measuring the permeability of the model pipe by adopting a permeability measuring device, and selecting the model pipe which meets the permeability. Vacuumizing saturated formation water, and calculating porosity; the parameters of the finally prepared model tubes for the subsequent steps are shown in table 2.
TABLE 2 model pipe Sand pack parameters
2. Continuously injecting petroleum from the inlet of the model pipe until the outlet of the model pipe detects the outflow of the petroleum, and measuring the original oil saturation; and aged at reservoir temperature (30 ℃) for 3 days.
3. And (4) displacing oil by using injected water, stopping water drive until the outlet produced liquid reaches the limit water content (98%), and calculating the recovery ratio (namely the water drive recovery ratio).
4. Injecting bacteria-containing water according to the injection amount of the volume of the pore of the 1 model tube (after the injection is completed, calculating the recovery ratio, namely the recovery ratio in the process of injecting the microorganisms), and culturing for 15 days at the oil reservoir temperature (30 ℃).
The preparation method of the bacteria-containing water comprises the following steps: inoculating activated W-Y14 in LB culture medium, performing shaking culture at 30 deg.C and 150rpm for 24h to obtain W-Y14 bacterial liquid (bacterial concentration is not less than 10)8cfu/mL). Adding W-Y14 bacterial liquid and NH into the injected water4H2PO4、(NH4)2SO4、K2HPO4、KNO3Peptone and yeast powder, wherein the concentrations of the peptone and the yeast powder in the bacteria-containing water are respectively 5% (volume fraction) W-Y14 bacteria liquid and 1g/L NH4H2PO4、1g/L(NH4)2SO4、1g/L K2HPO4、3g/L KNO32g/L peptone and 1g/L yeast powder.
5. And then, injecting water to displace oil, stopping water drive until the outlet produced liquid reaches the limit water content (98%), and calculating the recovery ratio (namely the recovery ratio of the subsequent water drive under the action of microorganisms).
The recovery ratios in different stages of water flooding, microbial injection, microbial action follow-up water flooding and the like are compared, and the results are shown in table 3. Indoor physical model oil displacement shows that the W-Y14 strain can improve the recovery ratio by 8.12 percent.
TABLE 3 recovery ratio
| Stage of material model | Recovery ratio (%) | Enhanced recovery (%) |
| Water drive | 43.57 | / |
| Microbial injection process | 47.88 | 4.31 |
| Follow-up water drive of microbial action | 51.69 | 8.12 |
Sequence listing
<110> Beijing Ruichi energy technology Limited
<120> one strain of Xanthomonas tenuis W-Y14 and application thereof
<160>1
<170>PatentIn version 3.5
<210>1
<211>1376
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
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ttgctctctg ggtggcgagt ggcggacggg tgaggaatac atcggaatct gcctatacgt 60
gggggataac gtagggaaac ttacgctaat accgcatacg tcctacggga gaaagcgggg 120
gatcgcaaga cctcgcgcgg atagatgagc cgatgtccga ttagctagtt ggcggggtaa 180
aggcccacca aggcgacgat cggtagctgg tctgagagga tgatcagcca cactggaact 240
gagacacggt ccagactcct acgggaggca gcagtgggga atattggaca atgggcgcaa 300
gcctgatcca gccatgccgc gtgggtgaag aaggccttcg ggttgtaaag cccttttgtt 360
gggaaagaaa tcctgtcgat taatacccga cggggatgac ggtacccaaa gaataagcac 420
cggctaactt cgtgccagca gccgcggtaa tacgaagggt gcaagcgtta ctcggaatta 480
ctgggcgtaa agcgtgcgta ggtggttcgt taagtctgat gtgaaagccc tgggctcaac 540
ctgggaattg cattggaaac tggcgggcta gagtacggta gagggtagtg gaattcctgg 600
tgtagcagtg aaatgcgtag agatcaggag gaacatccgt ggcgaaggcg actacctgga 660
ccagtactga cactgaggca cgaaagcgtg gggagcaaac aggattagat accctggtag 720
tccacgccct aaacgatgcg aactggatgt tgggtgcaat ttggcactca gtatcgaagc 780
taacgcgtta agttcgccgc ctggggagta cggtcgcaag actgaaactc aaaggaattg 840
acgggggccc gcacaagcgg tggagtatgt ggtttaattc gatgcaacgc gaagaacctt 900
acctggcctt gacatgtccg gaacccttga gagatcgagg ggtgccttcg ggaaccggaa 960
cacaggtgct gcatggctgt cgtcagctcg tgtcgtgaga tgttgggtta agtcccgcaa 1020
cgagcgcaac ccttgtcctt agttgccagc acgtaatggt gggaactcta aggagaccgc 1080
cggtgacaaa ccggaggaag gtggggatga cgtcaagtca tcatggccct tacggccagg 1140
gctacacacg tactacaatg ggaaggacag agggctgcga acccgcgagg gcaagccaat 1200
cccagaaacc ttctctcagt ccggatcgga gtctgcaact cgactccgtg aagtcggaat 1260
cgctagtaat cgcagatcag cattgctgcg gtgaatacgt tcccgggcct tgtacacacc 1320
gcccgtcaca ccatgggagt ttgttgcacc agaagcaggt agcctaaccg caagga 1376
Claims (10)
1. A strain of Xanthomonas rattan Luteimonas sp.W-Y14 with the preservation number of CGMCC No. 17152.
2. The application of the xanthomonas luteoimmunas sp, or the bacterial suspension thereof, or the culture solution thereof, or the fermentation product thereof, or the microbial inoculum containing the same in the petroleum recovery;
or, the application of the xanthomonas tenosynoviae sp, or the bacterial suspension thereof, or the culture solution thereof, or the fermentation product thereof, or the microbial inoculum containing the same in the preparation of petroleum products;
the luceimonas sp. of the xanthomonas rattan is luceimonas sp.W-Y14 CGMCC No.17152 of the xanthomonas rattan.
3. The application of the xanthomonas luteoimmunas sp, or the bacterial suspension thereof, or the culture solution thereof, or the fermentation product thereof, or the microbial inoculum containing the same in microbial oil recovery;
or, the application of the xanthomonas tenosynoviae sp, or the bacterial suspension thereof, or the culture solution thereof, or the fermentation product thereof, or the microbial inoculum containing the same in the preparation of the microbial oil recovery product;
the luceimonas sp. of the xanthomonas rattan is luceimonas sp.W-Y14 CGMCC No.17152 of the xanthomonas rattan.
4. The application of the xanthomonas luteoii Luteimonas sp, or the bacterial suspension thereof, or the culture solution thereof, or the fermentation product thereof, or the microbial inoculum containing the same in petroleum emulsification;
or, the use of the xanthomonas tenosynoviae sp, or a bacterial suspension thereof, or a culture solution thereof, or a fermentation product thereof, or a microbial inoculum containing the same in the preparation of petroleum emulsified products;
the luceimonas sp. of the xanthomonas rattan is luceimonas sp.W-Y14 CGMCC No.17152 of the xanthomonas rattan.
5. The application of the xanthomonas luteoii Luteimonas sp, or the bacterial suspension thereof, or the culture solution thereof, or the fermentation product thereof, or the microbial inoculum containing the same in petroleum degradation;
or, the use of the xanthomonas tenofovir sp, or a bacterial suspension thereof, or a culture solution thereof, or a fermentation product thereof, or a microbial inoculum containing the same in the preparation of petroleum degradation products;
the luceimonas sp. of the xanthomonas rattan is luceimonas sp.W-Y14 CGMCC No.17152 of the xanthomonas rattan.
6. The application of the xanthomonas luteoii Luteimonas sp, or the bacterial suspension thereof, or the culture solution thereof, or the fermentation product thereof, or the microbial inoculum containing the same in the treatment of petroleum pollution;
or, the application of the xanthomonas tenosynoviae sp, or the bacterial suspension thereof, or the culture solution thereof, or the fermentation product thereof, or the microbial inoculum containing the same in the preparation of products for treating petroleum pollution;
the luceimonas sp. of the xanthomonas rattan is luceimonas sp.W-Y14 CGMCC No.17152 of the xanthomonas rattan.
7. The application of the xanthomonas luteoimmunas sp, or the bacterial suspension thereof, or the culture solution thereof, or the fermentation product thereof, or the microbial inoculum containing the same in the petroleum pollution remediation;
or, the application of the xanthomonas tenosynoviae sp, or the bacterial suspension thereof, or the culture solution thereof, or the fermentation product thereof, or the microbial inoculum containing the same in the preparation of products for repairing petroleum pollution;
the luceimonas sp. of the xanthomonas rattan is luceimonas sp.W-Y14 CGMCC No.17152 of the xanthomonas rattan.
8. A product contains active ingredients of Luteimonas sp, or a bacterial suspension, a culture solution, a fermentation product or a microbial inoculum containing the Luteimonas sp;
the product has any one of the following functions 1) to 6):
1) oil recovery;
2) microbial oil recovery;
3) petroleum emulsification;
4) degrading petroleum;
5) treating petroleum pollution;
6) repairing petroleum pollution;
the luceimonas sp. of the xanthomonas rattan is luceimonas sp.W-Y14 CGMCC No.17152 of the xanthomonas rattan.
9. A method of microbial oil recovery comprising the steps of: adding Luteimonas sp or bacterial suspension thereof or culture solution thereof or fermentation product thereof or microbial inoculum containing the Luteimonas sp during oil extraction;
the luceimonas sp. of the xanthomonas rattan is luceimonas sp.W-Y14 CGMCC No.17152 of the xanthomonas rattan.
10. A method for treating and/or remediating petroleum pollution, comprising the steps of: processing petroleum pollutants by using Xanthomonas vine Luteimonas sp or bacterial suspension thereof or culture solution thereof or fermentation product thereof or microbial inoculum containing the same;
the luceimonas sp. of the xanthomonas rattan is luceimonas sp.W-Y14 CGMCC No.17152 of the xanthomonas rattan.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910355269.4A CN110079479B (en) | 2019-04-29 | 2019-04-29 | Tenebriomonas lutescens W-Y14 and application thereof |
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| CN103865820A (en) * | 2012-12-10 | 2014-06-18 | 中国石油天然气股份有限公司 | Luteimonas sp., and preparation method and application thereof |
| CN104371941A (en) * | 2013-08-15 | 2015-02-25 | 中国科学院大连化学物理研究所 | Luteimonas capable of degrading petroleum hydrocarbon, and its application |
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| CN103865820A (en) * | 2012-12-10 | 2014-06-18 | 中国石油天然气股份有限公司 | Luteimonas sp., and preparation method and application thereof |
| CN104371941A (en) * | 2013-08-15 | 2015-02-25 | 中国科学院大连化学物理研究所 | Luteimonas capable of degrading petroleum hydrocarbon, and its application |
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| Title |
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| Microbial enhanced oil recovery in Baolige Oilfield using an indigenous facultative anaerobic strain Luteimonas huabeiensis sp.nov;Ke et al.;《Journal of Petroleum Science and Engineering》;20180831;全文 * |
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