CN105567595A - Bacillus atrophaeus 5-2a and applications thereof - Google Patents
Bacillus atrophaeus 5-2a and applications thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of tertiary oil recovery utilizing microbes and metabolism thereof to improve the crude oil yield, and discloses a strain of bacillus atrophaeus 5-2a, which is preserved in China Center for Type Culture Collection. The preservation number of the strain is CCTCC M2014673. The strain can take urea as the nitrogen source to synthesize lipopeptide surface active substances, which can sustain a high temperature and high salt environment and have wide pH adaptability. The strain can well desorb the crude oil adhered on filter paper and fine sand. The invention also discloses an application of bacillus atrophaeus 5-2a in degrading crude oil or solid paraffin. The strain has a strong performance on degrading crude oil and solid paraffin, can largely reduce the viscosity, and has a strong performance of producing gas and acid. The effect is prominent.
Description
Technical field
The invention belongs to the technical field of tertiary oil recovery utilizing microorganism and meta-bolites thereof to improve oil recovery factor, be specifically related to atrophy genus bacillus 5-2a and application thereof.
Background technology
Oil is Nonrenewable resources, more adopts fewer.Worldwide, through once with secondary oil recovery after, in stratum, still the crude oil of residual 60% ~ 70% cannot extraction; Have many oil fields to enter the exploitation middle and later periods, exhausted ore deposit ratio increases year by year; Also have the recovery ratio in a considerable amount of viscous crude ore deposit very low; Hypotonic, deep layer, High Temperature High Pressure High water cut and little fault block oil reservoir etc. should not drive by heat and exploit with traditional methods such as chemical floodings.How to improve the tar productivity of above-mentioned low yield oil reservoir, making full use of various difficulty oil recovery and hide resource, is current problem demanding prompt solution.
Utilize microbial technique to improve oil production rate and be considered to the most promising current oil recovery technique, the depleted reservoirs exploitation for High water cut has even more important meaning.Microbial Enhanced Oil Recovery is the biology and the physical chemistry complex art that comprise the complex processes such as microorganism growth, migration and metabolism, has multiple action and net effect, but has stronger selectivity and specific aim.
Summary of the invention
For problems of the prior art, the object of the present invention is to provide strain atrophy genus bacillus 5-2a and an application thereof, this bacterium has very strong degraded and produces surfactant function, produce acid, aerogenesis simultaneously, viscosity of crude can be reduced, and the crude oil on attachment medium is come off, possess and improve crude oil fluidity and oil recovery factor several functions.
In order to reach above object, the present invention is achieved by the following technical solutions.
(1) atrophy genus bacillus 5-2a, be separated in the crude oil and petroleum-contaminated soil of Ansai Oilfield, Wuhan University of Wuhan City China typical culture collection center is preserved on December 28th, 2014, preserving number is CCTCCM2014673, called after atrophy genus bacillus 5-2a (Bacillusatrophaeus5-2a).
Reacted by PCR, from the 16SrDNA of genus bacillus 5-2a, amplification obtains the gene fragment of 1.6kb, after PCR primer purifying, is checked order by Nanjing Genscript Biotechnology Co., Ltd..By being analyzed with other bacterial strains 16SrDNA sequence in Genebank, the 16SrDNA sequence homology of this genus bacillus and BacillusatrophaeusAB021181 reaches 99.86%, therefore confirms that this genus bacillus 5-2a is atrophy genus bacillus (Bacillusatrophaeus).
Cultivated on beef-protein medium by atrophy genus bacillus 5-2a, this bacterial strain colony diameter is less, white, circular, bacterium colony is dry, and opaque, edge is irregular, bacterium colony surface folding, tarnish, under Electronic Speculum, cell is elongated rod shape, wide 0.5 ~ 1.0 μm, long 2.0 ~ 4.0 μm.Gram-positive, aerobic, glucose and sucrose ferment for positive, citric acid and malonate utilization are positive, and oxidation of ethanol is negative, and methyl red test and V-P test are the positive, nitrate reduction test and urease test are the positive, produce ammonia, denitrification test, can hydrolyzed starch and gelatin for negative.
(2) atrophy genus bacillus 5-2a can produce surfactant.
Further, described surfactant is lipopeptid class surfactant.
Further, described atrophy genus bacillus 5-2a can be nitrogenous source synthesis lipopeptid class surfactant with urea.
(3) application of atrophy genus bacillus 5-2a in oil degradation.
(4) application of atrophy genus bacillus 5-2a in solid paraffin degraded.
Utilize the mechanism of microbial technique raising oil production rate very complicated, be roughly divided into two aspects: one is the direct effect of microorganism cells to oil reservoir, if thalline is to the selectively blocking off effect of oil reservoir macropore and the Degradation to crude oil thereof, two is that microbial metabolites is as the indirect action to oil reservoir such as surfactant, biogas, acid and organic solvent.
Bio-surface active material be microorganism metabolism under certain condition synthesis there is the active and interfacial activity of certain surface, contain the amphoteric substance of hydrophilic group and hydrophobic group simultaneously, mainly comprise glycolipid class, lipopeptid class, phospholipid and in conjunction with polysaccharide etc., its type is relevant with bacterial strain and substrate.Bio-surface active material, except having reduction surface tension that synthetic surfactant has, stable emulsion and dispersion, solubilising and the performance such as wetting, has the advantages such as resource is easy to get, energy consumption is little, ecological safety simultaneously.Therefore, the existing genus bacillus by product surfactant is used for improving oil recovery factor at present, but institute's bacterium mostly is subtilis, Bacillus licheniformis and bacillus cereus etc., and selected bacterial strain or to produce surfactant, or based on the polymer substance in degrading crude oil, asynchronously have effect degraded and effectively produce the several functions such as surfactant, and there is no about the report of atrophy genus bacillus in the displacement of reservoir oil.
Compared with prior art, the present invention has the following advantages:
(1) atrophy genus bacillus 5-2a of the present invention can be nitrogenous source synthesis lipopeptid class surfactant with urea; This surfactant has good degreasing activity, emulsifying activity and lower surface tension value; Can the extreme environment such as withstand high temperatures, high salt, pH; To the crude oil that filter paper and fine sand adhere to, all there is good desorption.
(2) atrophy genus bacillus 5-2a of the present invention has degradation capability by force to crude oil and solid paraffin, and fall the feature that glutinous amplitude is large and aerogenesis acid producing ability is strong, action effect is remarkable.
The strong degradation function that above atrophy genus bacillus 5-2a has, aerogenesis function, produce acid function and produce all without reporting in the surfactant function document before the applying date, therefore this bacterium multi-functional displacement of reservoir oil bacterium that to be a strain new.This bacterium is respectively 37.8%, 22.0% and 53.1% to aromatic hydrocarbon, colloid and bitum degradation rate in crude oil; 30 DEG C, 40 DEG C time, make viscosity of crude comparatively contrast viscosity of crude and reduce by 24.7%, 25.6% respectively; 90.0%, 93.1% is respectively to the desorption of crude oil rate that fine sand, filter paper adhere to; Inhibiting rate for wax precipitation reaches 98.1%, and wax removal rate reaches 89.3%; The percent crystallization in massecuite of solid paraffin is made comparatively to contrast reduction by 88.9%; Gas production rate in crude oil is 2 times of fermentating liquid volume; Product acid amount in crude oil and solid paraffin culture system reaches 1.41g/L and 1.37g/L respectively, therefore this bacterium is to raising oil recovery factor, solves all have important using value to waxy crude oil production and a pipeline transportation wax deposition difficult problem.
Accompanying drawing explanation
Below in conjunction with the drawings and specific embodiments, the present invention is described in further details.
Fig. 1 is colonial morphology and the cell microscopic morphology figure of atrophy genus bacillus 5-2a.
Fig. 2 is the systematic evolution tree schematic diagram of atrophy genus bacillus 5-2a.
Fig. 3 is the infrared spectrogram of the Bio-surface active material that atrophy genus bacillus 5-2a produces.
Fig. 4 is the oil extraction loop diameter of differing temps, pH and salt concn process test strain fermented liquid, emulsification index and capillary comparison diagram.Wherein, the oil extraction loop diameter that Fig. 4-(a) is treatment of different temperature test strain fermented liquid, emulsification index and surface tension, the oil extraction loop diameter that Fig. 4-(b) is different pH process test strain fermented liquid, emulsification index and surface tension, the oil extraction loop diameter that Fig. 4-(c) is different salt concn process test strain fermented liquid, emulsification index and surface tension.
Fig. 5 is the solute effect comparison diagram of atrophy genus bacillus 5-2a process solid paraffin in normal hexane; Wherein, figure left side be blank group to solid paraffin solvability effect, figure the right side be atrophy genus bacillus 5-2a to the deliquescent effect of solid paraffin.
Fig. 6 is the desorption effect comparison diagram of atrophy genus bacillus 5-2a fermented liquid to filter paper, fine sand absorption crude oil; Wherein, Fig. 6-(a) is the desorption effect comparison diagram to Adsorption of Filter Paper crude oil, and a figure left side is the blank group of desorption effect to filter paper, and figure is right for atrophy genus bacillus 5-2a fermented liquid is to the desorption effect of filter paper; Fig. 6-(b) is the desorption effect comparison diagram to fine sand absorption crude oil, and a figure left side is the blank group of desorption effect to fine sand, and figure is right for atrophy genus bacillus 5-2a fermented liquid is to the desorption effect of fine sand.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in further details, but following examples are as limiting the scope of the invention.
Embodiment 1 bacteria selection
Utilize enrichment culture and dilution-plate method to separate several strain crude oil and paraffin degradation bacteria from the crude oil and petroleum-contaminated soil of Ansai Oilfield, Zhidan oil field, filter out high yield surfactant bacterial strain further by oil extraction circle and measurement of surface tension method.
The qualification of embodiment 2 bacterial classification
(1) morphological specificity
Cultivated on beef-protein medium by atrophy genus bacillus 5-2a, this bacterial strain colony diameter is less, white, circular, bacterium colony is dry, and opaque, edge is irregular, bacterium colony surface folding, tarnish, under Electronic Speculum, cell is elongated rod shape, wide 0.5 ~ 1.0 μm, long 2.0 ~ 4.0 μm.Concrete form is as Fig. 1.
(2) biochemical characteristic
Gram-positive, aerobic, glucose and sucrose ferment for positive, citric acid and malonate utilization are positive, and oxidation of ethanol is negative, and methyl red test and V-P test are the positive, nitrate reduction test and urease test are the positive, produce ammonia, denitrification test, can hydrolyzed starch and gelatin for negative.
(3) 16SrDNA sequential analysis
Reacted by PCR, from the 16SrDNA of genus bacillus 5-2a, amplification obtains the gene fragment of 1.6kb, after PCR primer purifying, is checked order by Nanjing Genscript Biotechnology Co., Ltd..By being analyzed with other bacterial strains 16SrDNA sequence in Genebank, the 16SrDNA sequence homology of this genus bacillus and BacillusatrophaeusAB021181 reaches 99.86%, therefore confirms that this genus bacillus 5-2a is atrophy genus bacillus (Bacillusatrophaeus).The systematic evolution tree of atrophy genus bacillus 5-2a is as Fig. 2.
Embodiment 3 bacterial classification produces surfactant
(1) bacterial classification produces the qualification of surfactant
Authentication method: first adopt acidization precipitation surface active substance, then carry out qualitative analysis with thin layer chromatography, by FTIR spectrum analysis, further qualification is done to its functional group.Specifically comprise the following steps:
1. the preparation method of atrophy genus bacillus 5-2a fermented liquid is: be linked on atrophy genus bacillus inclined-plane and be equipped with in the 600mL tissue culture bottle of 100mL liquid fermentation medium, 120r/min, and 3d, atrophy genus bacillus 5-2a fermented liquid cultivated by 30 DEG C of shaking tables.Wherein, the composition of liquid fermentation medium is: MgSO
47H
2o0.3, KH
2pO
45.0, K
2hPO
43H
2o10.0, NaCl5.0, urea 3.0, edible brown sugar 10.0, tap water 1000mL, pH7.0.
2. acidization precipitation surface active substance: get atrophy genus bacillus 5-2a fermented liquid supernatant liquid 10ml, about 6mol/LHCl adjust pH to 2.0 ~ 3.0, put into 4 DEG C, refrigerator and spend the night, observes whether adularescent precipitation produces.If there is white precipitate, show in fermented liquid containing the lipopeptid class surface active ingredient that bacterial metabolism is formed; If produce without white precipitate, then show in fermented liquid containing the glycolipid class surface active ingredient that bacterial metabolism is formed.The fermented liquid of centrifugal adularescent precipitation, the centrifugal 10min of 8000r/min, collect centrifugal gained precipitation, 1mol/L salt acid elution 2 times, obtains surfactant crude product, 40 DEG C of oven dry.
3. thin-layer chromatography: claim surfactant crude product 5mg, be hydrolyzed 24h at 6mol/LHCl110 DEG C, carry out thin-layer chromatography.Coloration method: chromatography terminates rear blower and dried up by thin plate, puts into 110 DEG C of baking ovens and is incubated 10min after spraying developer.Glycolipid developer is phenolsulfuric acid reagent, shows brown spot with glycolipid effect; Lipopeptid developer is triketohydrindene hydrate developer, aobvious red with lipopeptid effect.
4. functional group's qualification: FTIR spectrum analysis, carries out with the Tensor27 type infrared spectrometer that BRUKER company produces.Adopt KBr pressed disc method, sweep limit 400-4000cm
-1.
Qualification result: through thin-layer chromatography and infrared spectrum, the surfactant that bacterial strain 5-2a produces is fat peptide matters, and its infrared spectra spectrogram as shown in Figure 3.
(2) surfactant factors affecting stability:
Test method: after fermentation culture terminates, removes thalline by centrifugal for fermented liquid 8000r/min 10min, and collect supernatant liquor and press following routine processes, process terminates the oil extraction loop diameter of rear mensuration fermented liquid 20 DEG C time, emulsification index and surface tension.
A) temperature stability: get 20mL supernatant liquor, 6h at putting 20,30,40,50,60,80,100 and 120 DEG C respectively.
B) pH stability: get 20mL supernatant liquor, adjusts pH to 2.0,3.0,4.0,5.0,6.0,7.0,8.0,9.0,10.0,11.0,12.0,13.0 respectively with 1mol/LNaOH and HCl, acts on 24h at 20 DEG C.
C) salt concn stability: get 20mL supernatant liquor, with NaCl adjust salt concn 0 respectively, 10,30,50,70,90,110,150,200,300,500,700g/L, act on 24h at 20 DEG C.
Test-results: as shown in Figure 4, from Fig. 4-(a), temperature increases produces the degreasing activity impact of surfactant not quite to this bacterium institute.Between 20 ~ 120 DEG C, the oil extraction loop diameter for the treatment of of different temperature fermented liquid, emulsification index and surface tension value fluctuating range are little, illustrate this bacterium produce surfactant there is good temperature stability; From Fig. 4-(b), when pH value is 6.0 ~ 13.0, the change of the oil extraction loop diameter of different pH process fermented liquid, emulsification index and surface tension value is little, relatively stable; When pH value is below 6.0, due to Surfactant pellets, oil extraction loop diameter and emulsification index all reduce, surface tension increase, show strains tested produce surfactant in the basic conditions compared with there being higher stability under acidic conditions.From Fig. 4-(c), when salt concn is less than 130g/L, the emulsification index of fermented liquid and surface tension value relatively stable, oil extraction loop diameter slightly declines; During salt concn 130 ~ 300g/L, oil extraction loop diameter and the emulsification index of fermented liquid decline fast, and surface tension value increases; When salt concn is for being greater than 300g/L, fermented liquid still has certain degreasing activity, but its emulsifying property disappears.
The application of embodiment 4 bacterial classification
(1) to the degraded of crude oil
Test method: add 2.000g crude oil respectively in the thin mouth vial of 100mL, then add 30mL atrophy genus bacillus 5-2a fermented liquid, to add 30mL distilled water for contrast, add bottle closure of rubber sealing blend compounds band and reinforce, in case pressure of the inside of a bottle increases wash bottle stopper open.40 DEG C of standing and reacting 96h, shake in every 4 hours 1 time.After reaction terminates, measure after atrophy genus bacillus 5-2a degrades, stable hydrocarbon, aromatic hydrocarbon, colloid and asphalt content change in crude oil.
Test-results: as shown in table 1, as seen from table, genus bacillus 5-2a is respectively 12.5%, 37.8%, 21.7% and 53.1% to stable hydrocarbon, aromatic hydrocarbon, colloid and bitum degradation rate in crude oil, and atrophy genus bacillus 5-2a degradation treatment crude oil all reaches conspicuous level (P < 0.05) with the difference contrasting 4 kinds of components in crude oil.
Table 1 atrophy genus bacillus 5-2a is on the impact of crude oil group composition
(2) to the dissolving of Crude viscosity and solid paraffin
The dissolving situation of atrophy genus bacillus 5-2a to Crude viscosity and solid paraffin is as shown in table 2, and as seen from table, when 30 DEG C, after atrophy genus bacillus 5-2a degradation treatment, Crude viscosity comparatively control group Crude viscosity reduces by 24.7%; When 40 DEG C, comparatively control group reduces by 25.6%.Solid paraffin is after atrophy genus bacillus 5-2a degrades, solvability generation noticeable change in normal hexane, its solute effect as shown in Figure 5, solubilized paraffin amount has reached 68.6% of paraffin total amount, undissolved crystallization paraffin amount accounts for paraffin total amount 25.3%, all reaches conspicuous level (P < 0.05) with control group difference.This result shows, atrophy genus bacillus 5-2a significantly can reduce viscosity of crude, and can improve the dissolving power of paraffin in normal hexane, increases the small molecules paraffin amount that normal hexane is solvable.
Table 2 atrophy genus bacillus 5-2a is on Crude viscosity and the deliquescent impact of solid paraffin
(3) gas production rate of crude oil and solid paraffin being degraded and gaseous constituent
The gas production rate that atrophy genus bacillus 5-2a degrades to crude oil and solid paraffin is as shown in table 3, and as seen from table, have gas to produce in atrophy genus bacillus 5-2a degrading crude oil and solid paraffin process, through gas chromatographic analysis institute, aerogenesis body is mainly H
2and CO
2.As seen from Table 3, in degradation process, total gas production reaches 55.5 ~ 63.7mL/ bottle, wherein, and H
2be 36.0 ~ 41.5mL/ bottle, CO
2output be 19.5 ~ 22.2mL/ bottle, factor of created gase is 64.9% ~ 212.3%.
Aerogenesis kind and gas production rate in table 3 atrophy genus bacillus 5-2a degrading crude oil and solid paraffin process
(4) the product acid that crude oil and solid paraffin are degraded is measured and sour composition
Atrophy genus bacillus 5-2a measures as shown in table 4 to the product acid that crude oil and solid paraffin are degraded, as seen from table, after atrophy genus bacillus 5-2a degrades, reaction solution pH comparatively contrasts decline 26.5% ~ 27.1%, total acid content is 22.9 ~ 23.5mmol/L, and acid number reaches 1374 ~ 1410mg/L.Through organic acid stratographic analysis, short chain organic acid mainly oxalic acid and the propionic acid of generation.
Nutrient solution pH when table 4 atrophy genus bacillus 5-2a degrading crude oil and solid paraffin and produce acid measure
(5) fermented liquid degreasing activity and the desorption to crude oil
Test method:
A fermented liquid degreasing activity: get the plastic tub that bore is 24cm, add water near full, the water surface drips 2 drop of liquid paraffin, treat that paraffin free diffusing on the water surface forms 1 layer of whiteruss film, 1 fermented liquid to be measured is added at cere center, diffuse to form after circular row scraper ring towards periphery until Parafilm, measure oil extraction loop diameter (D) with ruler.Bio-surface active component content in D value and fermented liquid and degreasing activity thereof are proportionate, and are treated to contrast (CK) to drip distilled water.
B is to the desorption of crude oil: desorption of crude oil rate when being 1. absorption carrier with filter paper: immersed in crude oil by 6.0cm × 6.0cm qualitative filter paper of having weighed, suction after crude oil weigh until filter paper; The filter paper of absorption crude oil is placed in the 100mL wide-mouth tissue culture bottle that 80mL ferment product is housed, to add 80mL distilled water for contrast, 40 DEG C of quiescent culture 48h, weigh after taking out filter paper natural air drying, the tack of observed and recorded crude oil on filter paper is also taken pictures, and reckoner fermented liquid is to the desorption rate of crude oil.2. take particle diameter 0.25 ~ 0.50mm fine sand 90.0g to join in 600mL wide-mouth tissue culture bottle, add 10.0g crude oil subsequently, mix evenly, room temperature places 48h.Add 100mL test strains fermented liquid, be treated to contrast to add 100mL distilled water, 40 DEG C of quiescent culture 24h.The part crude oil disengaging grains of sand that reaction terminates rear grains of sand surface attachment are gathered in aqueous phase surface formation oil reservoir; Tissue culture bottle is placed in 4 DEG C of refrigerators and leave standstill, tissue culture bottle is taken out after crude oil solidifies, the state crude oil that solidifies of aqueous top layer in bottle is taken out, the cell free fermentation liquid of state crude oil surface attachment is solidified with tap water cleaning, blot residual moisture with thieving paper to weigh, calculate fermented liquid to the desorption rate of crude oil according to the crude quality come off from the grains of sand.
Test-results: as shown in table 5, as seen from table, this atrophy genus bacillus 5-2a fermented liquid has stronger surfactivity, and its oil extraction loop diameter is 19.0cm, is 63.3 times of contrast; This fermented liquid all has good desorption to the crude oil that fine sand, filter paper adhere to, it adsorbs the desorption effect figure of crude oil respectively as Fig. 6-(a), Fig. 6-(b) to filter paper, fine sand, in the desorption test process taking fine sand as adsorption medium, the crude oil of 90.0% splits away off from fine sand, and the amount of coming off is 9.7 times of contrast; In the desorption test process taking filter paper as adsorption medium, the crude oil of 93.1% splits away off from filter paper, and the amount of coming off is 7.3 times of control group.
Table 5 atrophy genus bacillus 5-2a fermented liquid oil extraction loop diameter and the impact on desorption of crude oil rate
(6) wax removal rate and inhibiting rate for wax precipitation
Test method:
A wax removal rate: adopt hanging slice method.Take slide glass as crude oil absorption carrier, measure the wax removal rate of ferment product.Immersed in crude oil by 26mm × 76mm slide glass of having weighed, make crude oil covered with slide glass lower end 5.3cm length inner area, retaining slide glass upper end 2.3cm is highly without oily district, weighs in the balance and be heavily accurate to 0.001g after crude oil solidifies; The slide glass scribbling crude oil is placed in the 600mL wide-mouth tissue culture bottle that 100mL fermented liquid is housed, to add 100mL distilled water for contrast, 30 DEG C of quiescent culture 96h; Take out slide glass, with the thalline that the clean slide glass of clear water rinsing adheres to and impurity, weigh.Before and after fermented liquid effect, the of poor quality of slide glass is the remaining paraffin deriving from crude oil, calculates wax removal rate (%).
B inhibiting rate for wax precipitation: adopt stationary method.Accurately taking 2.000g waxy crude oil is respectively charged in 100mL medical brine bottle, then add 30mL ferment product respectively, 30 DEG C of constant temperature culture 48h, cultivate and pour in 50mL small beaker by the mixture of fermented liquid and crude oil after terminating, by by clean-out system and washed with de-ionized water and dry slide glass, vertical hanging is in the fermented liquid containing crude oil, in 1h, crude oil temperature is down to 25 DEG C by 60 DEG C, take out slide glass after leaving standstill 30min, wash away crude oil with Virahol, then weigh.To add 30mL distilled water for blank.Calculate bacterium inhibiting rate for wax precipitation (%).
Test-results: as shown in table 6, as seen from table, this atrophy genus bacillus 5-2a has good wax removal, wax control effect.Wherein, inhibiting rate for wax precipitation reaches 98.1%, and wax removal rate reaches 89.3%.
Table 6 atrophy genus bacillus 5-2a lab simulation Paraffin Removal rate
Comprehensive above embodiment is known, the atrophy genus bacillus 5-2a of the present invention's screening has following multiple displacement of reservoir oil function: 1. Degradation: can alkane, aromatic hydrocarbon, colloid and pitch significantly in degrading crude oil and solid paraffin, the heavy component in crude oil and paraffin is made to be decomposed into light component, reduce Crude viscosity, the paraffin amount that minimizing can deposit, and then effectively improve the mobility of crude oil and the wax crystal particle of paraffin.2. aerogenesis: bacterial strain produces a large amount of H in degrading crude oil and paraffin process
2and CO
2, these gases can increase oil reservoir internal pressure; Gas dissolves in crude oil can reduce Crude viscosity, improves crude oil fluidity.3. acid is produced: in degrading crude oil and paraffin process, produce the organic acids such as a large amount of oxalic acid, propionic acid, effectively can dissolve the carbonate in oil reservoir rock hole, increase the porosity and permeability of oil reservoir, the interfacial tension between profit can also be reduced simultaneously, form oil-water emulsion, thus improve oil recovery factor.4. surfactant is produced: this bacterial strain can be nitrogenous source synthesis lipopeptid class surfactant with urea, this surfactant can withstand high temperatures, high salt, there is pH adaptability more widely, and to filter paper, fine sand absorption crude oil, all there is good desorption, contribute to desorption of crude oil, to the paraffinic components in crude oil, there is certain emulsification, dissemination simultaneously, wax recovery is attenuated and diminishes and flow out oil well with Produced Liquid, improve wax control wax removal effect, and then improve oil recovery factor.
Known by inference by above each effect, this multi-functional displacement of reservoir oil sporeformer, to improving the effect of oil recovery factor, wax removal and wax control by being better than active substance producing strains or the degradation bacteria with simple function, has wide application potential in crude oil exploration industry and pipeline transportation.
Although the present invention is described in detail with a general description of the specific embodiments in this specification sheets, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, all belong to the scope of protection of present invention.
Claims (6)
1. atrophy genus bacillus (Bacillusatrophaeus) 5-2a, is preserved in China typical culture collection center, and preserving number is CCTCCM2014673.
2. atrophy genus bacillus 5-2a according to claim 1, is characterized in that, the 16SrDNA sequence of described atrophy genus bacillus 5-2a is as shown in SEQIDNo.1.
3. atrophy genus bacillus 5-2a according to claim 1, is characterized in that, described atrophy genus bacillus 5-2a can produce surfactant; Described surfactant is lipopeptid class surfactant.
4. atrophy genus bacillus 5-2a according to claim 3, is characterized in that, described atrophy genus bacillus 5-2a can be nitrogenous source synthesis lipopeptid class surfactant with urea.
5. the application of atrophy genus bacillus 5-2a in oil degradation described in claim 1 or 2.
6. the application of the atrophy genus bacillus 5-2a described in claim 1 or 2 in solid paraffin degraded.
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