CN101092602A - Viscosity reduction bacteria for increasing recovery ratio of petroleum, and application - Google Patents
Viscosity reduction bacteria for increasing recovery ratio of petroleum, and application Download PDFInfo
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- CN101092602A CN101092602A CNA2007100906957A CN200710090695A CN101092602A CN 101092602 A CN101092602 A CN 101092602A CN A2007100906957 A CNA2007100906957 A CN A2007100906957A CN 200710090695 A CN200710090695 A CN 200710090695A CN 101092602 A CN101092602 A CN 101092602A
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Abstract
This invention discloses Streptococcus zooepidemicus (CGMCC No.1988) for reducing petroleum viacosity and increasing petroleum yield, and its application. Streptococcus zooepidemicus is applied by: inoculating in petroleum, and fermenting at 20-60 deg.C for 8-72 h. Streptococcus zooepidemicus can reduce petroleum viacosity from 8000-15000 mPa.s to 50-250mPa.s, increase petroleum acid value by 8-15 times and light component contents, reduce wax and glue contents by 50-95%, improve rheological property of petroleum, reduce interfacial tension between water and oil, increase organic acid contents in fermented petroleum, and increase oil layer pressure and petroleum fluidity. Besides, CO2, biogas and hyaluronic acid are generated during the fermentation process, thus fermented petroleum is easy to emulsify, moisten and disperse. Streptococcus zooepidemicus can be used in oil production, especially microbe-enhanced oil production and petroleum transportation.
Description
Technical field
The present invention relates to a strain improves oil recovery in the bioengineering field viscosity reduction microorganism and application thereof, particularly a strain improves the viscosity reduction bacterium and the application thereof of oil recovery.
Background technology
The world today is carrying out one with biotechnology and be applied as the new technology revolution of sign, and biological develop energy technology becomes economic potential maximum in the biotechnology, is hopeful one of field with future most, and these factors have promoted microbial enhanced oil recovery Study on Technology and application.
In the petroleum industry, primary oil recovery is that crude oil relies on subsurface pressure at present, and from the process that sprays ground, along with the reduction of subsurface pressure, blowing finishes by oil well, but about the 10-15% of the underground crude oil of primary oil recovery extraction; Secondary oil recovery is by toward underground continuous water filling, replenishes underground energy, thereby realizes the purpose of water displacing oil.But along with the increase of water injection rate, water content constantly rises in the extraction liquid, and is moisture up to about 92-94% in the present Chinese onshore oil field extraction liquid.According to internationally recognized standard, moisturely in the extraction liquid reach 97%, both there has not been economic benefit, but secondary oil recovery also only about 20% of the underground crude oil of extraction.
Oil is a kind of deregenerative energy, and after primary oil recovery and secondary oil recovery, the crude oil of the 60-70% that still has an appointment in the stratum can't be exploited out.How to improve recovery ratio, from the more crude oil of underground extraction, be the constantly problems of research of many countries for many years always.What generally adopt at present is to use the chemical process crude oil extraction, as the ASP ternary composite oil-displacing system of alkali, tensio-active agent, polymkeric substance composition.But there is shortcoming in this method, because generally all be higher than 50 ℃ at subsurface temperature, and polyacrylamide is under the sodium hydroxide existence condition, 50 ℃ just are degraded to small molecules or unit molecule acrylamide very soon, and acrylamide has not only lost the oil displacement efficiency of polyacrylamide, and be toxic substance, a large amount of uses will cause phreatic pollution.Since 1980, a lot of countries have carried out the technology that improves oil recovery factor with microbial process, and through the effort of two more than ten years, this technology has obtained very big progress.This use microorganism is improved recovery efficiency technique and is called four oil recovery techniques or microbial enhanced oil recovery technology, is meant and utilizes the useful product of microorganisms producing or utilize microorganism can decompose the performance of hydrocarbon polymer, thereby improve the technology of oil recovery factor.The microbial enhanced oil recovery technology is a new technology high in technological content, that development is swift and violent, is modern biotechnology ground-breaking application in the petroleum production engineering field, more demonstrates its great vitality for the moisture and approaching exhausted maturing field of height.
The microbial enhanced oil recovery technology mainly comprises two classes: a class is to utilize microniological proudcts such as biological polymer xanthan gum and bio-surfactant to carry out the displacement of reservoir oil as the oilfield chemistry agent, be called microorganism ground top fermentation and improve recovery ratio technology, be the bioprocess technology method, this technology at home at present, oneself becomes ripe outward; Another kind of is to utilize microorganism and meta-bolites thereof to improve recovery ratio, mainly is the vigor that utilizes microorganism ground bottom fermentation and utilize autochthonous microorganism in the oil reservoir, is called microorganism ground bottom fermentation and improves the recovery ratio method.
The separation screening that improves the viscosity reduction microorganism of oil recovery is the basis of realizing the microbial enhanced oil recovery technology, and the quality of bacterial classification is the key of microbe oil production.At present, separated the microorganism that obtains some degraded oils, reported the experiment that discharges crude oil with sulphate reducing bacteria, studies show that the good result of sulphate reducing bacteria performance in the bacterium oil decomposes as nineteen forty-seven U.S. scientist Beckman.Nineteen fifty-five, dimension thunder in Larry was observed the surface tension in cultures such as glycerine such as Pseudomonas aeruginosa, mycobacterium, withered grass bud pole bacterium, and surface tension descends obviously, but utilizes hydro carbons and bad.Ni Yin utilized Corynebacterium, mycobacterium and Nocardia as emulsion splitter in 2002, can reduce oil water interfacial tension but must combine with chemical demulsifier and inorganic salt, and application suffers restraints.Hou Zhao in 2004 is big filter out cured shape genus bacillus and short genus bacillus rheological characteristic of crude oil improve aspect and between profit interfacial tension descend certain effect all arranged, but effect is unsatisfactory under the higher condition of temperature.
Summary of the invention
The purpose of this invention is to provide viscosity reduction bacterium and application thereof that a strain can improve oil recovery.
Streptococcus zooepidemicus (Streptococcus zooepidemicus) the CGMCC NO.1988 that the present invention is used, in China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation, preserving number is streptococcus zooepidemicus (Streptococcus zooepidemicus) CGMCC NO.1988, and the preservation time is on 03 28th, 2007.
Streptococcus zooepidemicus (Streptococcus zooepidemicus) CGMCC NO.1988 separates to obtain from the nasal mucosa of ox, and its bacterium colony is that white is transparent, moistening, and diameter 1.0-5.0mm, cell are that spherical shape or chain are spherical; Cell dia≤2 μ m; Cell peripheral band pod membrane, pod membrane are excretory hyaluronic acid biological polyoses in the cell cultivation process meeting; Concrete physiological and biochemical property is as shown in table 1:
Table 1. streptococcus zooepidemicus CGMCC NO.1988 physiological and biochemical property
| Test subject | The result | Test subject | The result |
| Carbohydrate produces sour sucrose glucose utilization sucrose aerogenesis and utilizes the maltose aerogenesis to utilize the glucose aerogenesis to utilize sucrose product polysaccharide to utilize maltose product polysaccharide to utilize glucose product polysaccharide to utilize phosphate to utilize 80 ℃ of survivals of 60 ℃ of growths of sulfate pH3.0-9.5 to grow | + + + + + + + + + + + + + + | The facultative growth of casein methyl red test |
Positive+++---+++++++ |
Annotate: "+" expression growth or reacting positive; "-" expression is not grown or reaction negative.
Experimental results show that, streptococcus zooepidemicus (Streptococcus zooepidemicus) CGMCC NO.1988 can improve the character of crude oil effectively, viscosity of crude drops to 50-250mPas by 8000-15000mPas, the 60-160 that descended doubly, acid value for crude oil has improved more than 8-15 times, light component increases, the content of wax contains glue and is reduced to 4-10% by 20-45%, reduced rate is 50-95%, rheological improves, interfacial tension reduces between profit, and substances content such as organic acid increases in the fermented liquid, and fermenting process produces biogas CO
2Its metabolism crude oil approach is time terminal oxidation, has produced a kind of biological polyoses.This bacterial strain can be widely used in petroleum production engineering field and oil transport field, particularly microbial enhanced oil recovery field.
The screening of streptococcus zooepidemicus (Streptococcus zooepidemicus) CGMCC NO.1988, form by following process steps:
(1) sampling and enrichment culture;
(2) screening of streptococcus zooepidemicus and purifying;
(3) bacterial strain after will screening carries out complex mutation;
(4) will use gammairradiation and magnetic field treatment through the streptococcus zooepidemicus of complex mutation;
(5) high temperature acclimation of streptococcus zooepidemicus.
Concrete processing step and processing condition are:
Step (1) sampling and enrichment culture:
From the nose of an ox of tens of oxen newly slaughtering, scrape and get active strong nasal mucosa, carry out enrichment culture, obtained sticking bacterial strain.Sticking bacterial strain fermentation liquor extraction product and the contrast of hyaluronic acid standard substance infared spectrum are analyzed, determined that starting strain is a streptococcus zooepidemicus.
Enrichment medium is made up of following parts by weight of component:
Sucrose 10-40 part, peptone 20-60 part, extractum carnis 10-50 part, (NH
4)
2SO
42-8 part, sal epsom 0.5-2 part, sodium-chlor 2-5 part, K
2HPO
42-5 part, NaH
2PO
42-5 part, CaCl
22H
20.05 part of O, FeCl
24H
21000 parts in 0.1 part of O and water;
The processing condition of preparation enrichment medium are: pH value is 6.0-8.0, sterilizes 20 minutes for 121 ℃, and culture temperature is that 25-60 ℃, incubation time are 8-48 hour.
The screening and the purifying of step (2) bacterial classification:
The selective medium of screening usefulness is a crude oil inorganic salt solid plate substratum.
Do not add any carbon source in this selective medium except crude oil, the bacterial classification that filters out is exactly can be the bacterial classification of sole carbon source growth with crude oil.Crude oil after bacterium liquid and the dilution is mixed, pour on the inorganic salt solid medium flat board for preparing, after the crude oil of waiting to contain bacterium liquid is tiled in the inorganic salt planar surface equably and solidifies.Cultivated 8-48 hour for 25-60 ℃, the result can find out obviously that the local crude oil long bacterium is utilized as shown in Figure 1, and the former oil reservoir attenuation on the inorganic salt flat board forms transparent circle.Select to form the bigger bacterial strain of transparent circle, carry out complex mutation.
Crude oil inorganic salt solid plate substratum is made up of following parts by weight of component:
Crude oil 10-400 part, peptone 20-60 part, extractum carnis 10-50 part, (NH
4)
2SO
42-8 part, sal epsom 0.5-2 part, sodium-chlor 2-5 part, K
2HPO
42-5 part, NaH
2PO
42-5 part, CaCl
22H
2O0.05 part, FeCl
24H
21000 parts in 0.1 part of O, 15 parts in agar and water;
The processing condition of preparation crude oil inorganic salt solid plate substratum are: the pH value is 6.0-8.0, sterilizes 20 minutes for 121 ℃, and culture temperature is 25-60 ℃, and incubation time is 8-48 hour.
The complex mutation of step (3) bacterial classification:
Complex mutation comprises the preparation of seed bacteria suspension successively, the add-on of mutagenic compound, time of ultraviolet irradiation.
The inoculation that step (2) is obtained is to containing in the 5ml physiological saline test tube, adding volume percent in the test tube is the ethyl sulfate mutagenic compound of 0.5%-2.0%, 1-2min vibrates on the vortex oscillation device, get the crude oil inorganic salt solid plate of this bacteria suspension application step (2) of 0.5ml, flat board is placed on ultra violet lamp 20-60s, then flat board is secretly cultivated.Culture condition is: culture temperature 25-60 ℃, incubation time is 24-72 hour.
The gammairradiation and the magnetic field treatment of step (4) bacterial classification:
The cultured bacterial classification flat board of step (3) is placed on irradiation (gamma-rays dosage is 12.9-38.7 C/kg) under the gamma-rays, postradiation flat board is handled (magneticstrength is 10-100T) with magnetic field again, at last the flat board after handling being put into incubator cultivates, culture condition is: culture temperature 25-60 ℃, incubation time is 24-72 hour.
The high temperature acclimation of step (5) bacterial classification:
Will be through the crude oil inorganic salt solid plate substratum of the bacterial strain application step (2) of mutagenesis screening, putting into 40 ℃ of incubators cultivated 8-48 hour, the bacterial strain of growth very fast (former oil reservoir transparent circle is bigger) is picked out the crude oil inorganic salt solid plate substratum that continues application step (2), putting into 50 ℃ of incubators cultivated 8-48 hour, still bacterial strain picks out the crude oil inorganic salt solid plate substratum that continues application step (2) growing faster, put into 60 ℃ of incubators and cultivated 8-48 hour, the picking bacterial strain of growing is faster preserved.
Obtain adapting to pyritous streptococcus zooepidemicus (Streptococcus zooepidemicus) CGMCCNO.1988 at last, be tested and appraised, this Pseudomonas is non-human pathogen in Lancefield serogroup C type streptococcus streptococcus zooepidemicus.
The obtained technical progress of the present invention is:
(1) bacterial classification vitality of the present invention is strong, all can well grow under environment such as high temperature, anaerobism, aerobic, facultative, high salt concentration, and thalline directly can grow in crude oil system, need not additionally to add other medium components, reduces cost for oil production greatly;
(2) bacterial classification of the present invention can be secreted the biological polyoses hyaluronic acid in process of growth, the hyaluronic viscosity that increases water on the one hand that exists of biological polyoses, and the flowability of reduction water reduces fingering and too early water logging, improves sweep efficiency, increases and sweeps oily efficient; On the other hand, the biological polyoses hyaluronic acid has increased the profit mixing, helps profit and forms emulsion, makes the easier emulsification of crude oil, wetting and dispersion; Moreover, biological polyoses hyaluronic acid as the streptococcus zooepidemicus secretory product, be at cell peripheral with the pod membrane wrapped, this form can effectively protect somatic cells to exempt from the injury of external environment, so, thalline has stronger vitality, can grow under more extreme environment (as high temperature, high pressure, high salt etc.);
(3) bacterial classification of the present invention low-molecular-weight organic acid of meeting metabolism in process of growth, their energy dissolved carbon hydrochlorates increase porosity, improve rate of permeation; In addition, bacterial classification improves sand pressure in process of growth meeting release of carbon dioxide, reduces viscosity of crude, improves the crude oil flow ability.
(4) bacterial classification of the present invention not only can be secreted biological polyoses hyaluronic acid and low-molecular-weight organic acid in process of growth, but also can produce biogas CO
2, this is different from the bacterial classification that in the past is used for degraded oil, is used for the bacterial classification of degraded oil in the past or only can secretes biological polyoses (as xanthan gum), or only understand the low-molecular-weight organic acid of metabolism (as pyruvic acid), or only can produce biogas (as CO
2).And bacterial classification of the present invention integrates the three, and the biological polyoses hyaluronic acid of generation makes the easier emulsification of crude oil, wetting and dispersion, can also protect somatic cells; Metabolic organic acid can the dissolved carbon hydrochlorate, can change rock surface character and crude oil physical properties, the crude oil that is adsorbed on the porous rocks surface is released, and is easy to extraction ground; The biogas that produces can make the supercharging of oil reservoir part and reduce viscosity of crude, improves the crude oil flow ability, increases rate of permeation, crude oil is expanded, and volume increases, and helps displacing crude oil, increase output, the Jamin effect of bubble also can increase flow resistance simultaneously, improves and injects ripples and volume;
(5) bacterial classification accommodative ability of environment of the present invention is strong, is all fine growths of energy under the 20-60 ℃ of condition at pH3.0-9.5 and temperature;
(6) bacterial classification emulsified crude oil of the present invention is rapid, and bacterial classification adds crude oil and got final product emulsification, viscosity reduction in 8 hours, has improved oil-production efficiency greatly;
(7) utilize the crude oil of bacterial classification extraction of the present invention, oil product subsequent processes (breakdown of emulsion and desalination) is simple, greatly reduces oil product subsequent processes cost.
Embodiment
(1) growth curve
With streptococcus zooepidemicus (Streptococcus zooepidemicus) CGMCC NO.1988 direct inoculation in 100ml crude oil, in 20-60 ℃, 120-180r/min shaking table shaking culture, regularly measure the density (OD value) of thalline in the crude oil, with time is X-coordinate, the OD value is drawn its growth curve for ordinate zou, and the result as shown in Figure 2.By growth curve as can be seen, thalline begins to enter logarithmic phase at crude oil 4h, promptly enters stationary phase about 10h, and this shows that thalline breeds very soon in crude oil, help practical application.
(2) dispersion and emulsion effect
Getting 5ml streptococcus zooepidemicus (Streptococcus zooepidemicus) CGMCC NO.1988 bacterium liquid directly joins in the 100ml crude oil, 20-60 ℃, the 120-180r/min shaking table was cultivated 8-72 hour, the result as shown in Figure 3, by found that, the color of fermented liquid is obviously deepened, and crude oil is dispersed in the fermented liquid, form emulsion, and effect back crude oil has the advantages that not hang bottle; And the blank fermented liquid (not connecing bacterium) that compares with it, the layering still of crude oil and fermented liquid, and also crude oil is basically all attached on the bottle wall.By the emulsion microscopy after the fermentation being found all be wrapped in one deck oil around each thalline, the motion that makes crude oil be accompanied by thalline is dispersed in the fermented liquid, profit forms emulsion, thereby plays emulsification, wetting, the effect that disperses crude oil.
(3) crude oil total hydrocarbon color atlas before and after the streptococcus zooepidemicus CGMCC NO.1988 effect
Get the crude oil of fermentation after 8-72 hour in the step (1), do the analysis of crude oil total hydrocarbon according to a conventional method.Result such as table 2 and shown in Figure 4 can be seen from table 2 and Fig. 4, and this bacterial strain is some the high carbon number alkane in the degrading crude oil optionally, and long chain hydrocarbon content reduces relatively, and short hydrocarbon or low chain hydrocarbon content increase relatively, and the light component of crude oil increases; In addition, ∑ C
21 -/ ∑ C
22 +(C
21+ C
22)/(C
28+ C
29) increase of ratio, expression crude oil is migrated to light constituent compound direction by high-molecular weight compounds.Along with the relative minimizing of macromolecular compound content, the relative increase of light constituent compounds content, the flowability of crude oil improves.
The stable hydrocarbon chromatogram detected result of table 2. streptococcus zooepidemicus CGMCC NO.1988 fermentation back crude oil
| The sample title | Carbon number range | Main peak carbon | ∑C 21 -/∑C 22 + | (C 21+C 22)/(C 28+C 29) |
| Blank crude oil | nC9-C36 | nC23 | 0.675 | 1.78 |
| NO.1988 | nC9-C34 | nC16 | 2.40 | 3.38 |
Annotate: ∑ C
21 -Expression heneicosane and former each carbon number normal paraffin massfraction sum, %;
∑ C
22 +Expression n-docosane and later each carbon number normal paraffin massfraction sum, %;
C
21Expression heneicosane hydrocarbon massfraction, %;
C
22Expression n-docosane hydrocarbon massfraction, %;
C
28Represent positive octacosane hydrocarbon massfraction, %;
C
29Represent positive nonacosane hydrocarbon massfraction, %;
(3) viscosity of crude changing conditions
Getting 5ml streptococcus zooepidemicus (Streptococcus zooepidemicus) CGMCC NO.1988 bacterium liquid directly is added in the 100ml crude oil, 20-60 ℃, the 120-180r/min shaking table was cultivated 8-72 hour, detect viscosity under 30 ℃ of the different time crude oil with the rotor viscometer, the result as shown in Figure 5, the result shows that effect back rheological characteristic of crude oil obviously improves, and viscosity descends significantly.After fermentation 8-12 hour, viscosity of crude is reduced to 50-250mPas by the preceding 8000-15000mPas of effect, and viscosity has reduced 60-160 doubly, and viscosity of crude descends greatly.
(4) the crude oil content of wax and the variation that contains glue
Get the crude oil that has fermented 8-72 hour in the step (1), vapor-phase chromatography is carried out the crude oil content of wax, is contained glue mensuration routinely, the result is as shown in table 3, the wax of crude oil and gelationus content all have decline in various degree before and after the bacterial classification effect, wax content in crude oil is original 11.4% after this bacterial strain effect, and gel content is original 32.9%.This explanation streptococcus zooepidemicus can utilize paraffin in the crude oil as utilization of carbon source, thereby makes the viscosity degradation of crude oil.For further the checking streptococcus zooepidemicus can be with paraffin as carbon source, we replace oil as sole carbon source with whiteruss in minimal medium, found that thalli growth is good, with be that the carbon source situation is the same substantially with the oil, this proves absolutely that this bacterium can well utilize the paraffin in the crude oil to be used as carbon source.
The variation of crude oil wax and glue before and after the table 3. streptococcus zooepidemicus CGMCC NO.1988 effect
| The sample title | The content of wax/% | Viscosity break ratio/% | Contain glue/% | Glue rate/% falls |
| Blank crude oil | 40.5 | 88.6 | 21.7 | 67.1 |
| NO.1988 | 4.63 | 7.14 |
(5) variation of nonhydrocarbon in the crude oil
Get the crude oil that has fermented 8-72 hour in the step (1), the variation of non-hydrocarbon constituents and structure in this bacterial strain effect front and back crude oil of employing Infrared spectroscopy.Analytical results shows that after microbiological treatment, bigger variation has taken place the infrared spectra of nonhydrocarbon in the viscous crude.C-O key absorption peak newly occurred, occurred stronger aldehyde radical, the asymmetric vibration absorption peak of aromatic base ether simultaneously, shown that streptococcus zooepidemicus has oxygenizement to non-hydrocarbon component in the crude oil; C h bond outside sweep vibration absorption peak on the phenyl ring also occurred, illustrated that this bacterium may have the effect of condensation aromatic ring in the nonhydrocarbon opened.In addition, 1697～1377cm
-1Peak absorption intensity in the scope increases, and illustrates that this bacterium can make aromatic ring top side chain come off, and increases methyl in the nonhydrocarbon, methylene radical content.
By measuring the acid number of this bacterial strain effect front and back crude oil nonhydrocarbon, the result is as shown in table 4, the result shows that crude oil is after this bacterial strain effect, the acid number of crude oil nonhydrocarbon is increased to original 8-15 doubly, illustrate that this bacterial strain in the degradation process to the crude oil different components biooxidation reactions has taken place, generate organic acid, improved the acid number of crude oil.
Crude oil nonhydrocarbon acid number analytical results before and after the table 4. streptococcus zooepidemicus CGMCC NO.1988 effect
| The sample title | Nonhydrocarbon acid number/% | The acid number increase/doubly |
| Blank crude oil | 0.0975 | 14 |
| NO.1988 | 1.372 |
(7) stable hydrocarbon biological degradation Analysis on Mechanism in the crude oil
From above research as can be seen, this bacterial strain to the degraded of crude oil to be degraded to main path.Microorganism is different to the pathways metabolism of different hydrocarbons compound in the oil with mechanism.Stable hydrocarbon comprises normal paraffin, branched paraffin and naphthenic hydrocarbon.It has been generally acknowledged that under microbial process, straight-chain paraffin at first is oxidized to alcohol, the alcohol that comes from alkane is oxidized to corresponding aldehyde under the effect of alcoholdehydrogenase, and aldehyde then is oxidized to lipid acid by the effect of aldehyde dehydrogenase.General, oxidative pathway has single terminal oxidation, diterminal oxidation and inferior terminal oxidation, and it may be by way of being shown below:
(1) single terminal oxidation:
R-CH
2-CH
3+O
2→R-CH
2-CH
2-OH→R-CH
2-CHO→R-CH
2-COOH
(2) diterminal oxidation:
H
3C-(CH
2)
n-CH
3+O
2→H
3C-(CH
2)
n-CH
2OH→H
3C-(CH
2)
n-CHO→H
3C-(CH
2)
n-COOH→HOH
2C-(CH
2)
n-COOH→OHC-(CH
2)
n-COOH→HOOC-(CH
2)
n-COOH
(3) inferior terminal oxidation:
H
3C-(CH
2)
11-CH
3→H
3C-(CH
2)
10-CH(OH)-CH
3→H
3C-(CH
2)
10-COCH
3→H
3C-(CH
2)
9-CH
2-O-COCH
3→H
3C-(CH
2)
9-CH
2OH+CH
3COOH
Relative normal paraffin, branched paraffin embarrass Institute of Micro-biology's degraded, and the existence of side chain has strengthened the resistance to corrosion of alkane, and side chain is many more big more, and the difficulty that is degraded by microorganisms is big more.The oxidation of branched paraffin also can be subjected to the inhibition of normal paraffin oxygenizement.This bacterial strain shows that to the composition analysis result of oil degradation the carbon number of mixed fatty acid concentrates on C
2-C
20In the scope.This illustrates that this bacterial strain may be since a comparatively special approach to the bio-oxidation of crude oil---inferior terminal oxidation.Crude oil medium high carbon chain stable hydrocarbon is chain fatty acid in time terminal oxidation generates two kinds at first, carries out terminal oxidation and β-Yang Hua again.
The analysis of embodiment 2, streptococcus zooepidemicus CGMCC NO.1988 bacterial strain fermentation liquor
(1) the organic acid qualitative and quantitative analysis of fermented liquid
1. organic acid The qualitative analysis
Utilize day island proper Tianjin GC-14C gas-chromatography: FFAP quartz capillary column (0.250 * 30m), carrier gas is a high pure nitrogen, flow 1ml/min, 220 ℃ of temperature of vaporization chamber, column temperature adopts temperature programming, the beginning temperature is 90 ℃, and speed is 3 ℃/min, and outlet temperature is 200 ℃, sample size 1.0 μ l, carry out the organic acid qualitative test according to ordinary method, the color atlas of organic acid standard model such as Fig. 6 a, the retention time under chromatographic condition is as shown in table 5.Organic acid gas chromatogram such as Fig. 6 b and the 6c of crude oil sample before and after the streptococcus zooepidemicus effect.Containing three kinds of organic acids in the crude oil sample of streptococcus zooepidemicus effect back as seen from the figure is acetate, propionic acid and butyric acid.
The retention time of table 5. organic acid standard specimen
| Acetate | Propionic acid | Butyric acid | |
| Retention time/min | 1.865 | 2.073 | 2.357 |
2. organic acid quantitative analysis results
Measure pretreated sample 100ml, place distilling flask, add the acidifying of 7ml phosphoric acid, then distillation.When distilling flask is left small volume of solution, after the cooling, respectively adds 20ml distilled water more at twice and continue distillation a little.Among the NaOH and distillate, control PH is 8-9.After being concentrated into about 5ml with Rotary Evaporators, sample is changed in the 10ml volumetric flask, be acidified to PH≤3, constant volume, organic acid to be measured with HCl.This extracting method can extract various organic acids fully, and gas chromatographic analysis is used in the various acid that extract respectively, and peak area is inserted in various acidity scale directrix curves, can obtain the concentration of various acid.Concrete outcome is as shown in table 6:
Table 6. organic acid quantitative analysis results
| Acetate | Propionic acid | Butyric acid | Total acid | |
| Blank crude quality concentration (g/L) | 2.1 | 3.5 | 3.3 | 8.9 |
| Blank crude oil volumetric molar concentration (mmol/L) | 35 | 47 | 38 | 120 |
| Handle back oil sample mass concentration (g/L) | 18.7 | 37.2 | 24.6 | 80.5 |
| Handle back oil sample volumetric molar concentration (mmol/L) | 312 | 503 | 280 | 1095 |
(2) the organic pure qualitative and quantitative analysis of fermented liquid
Utilize day island proper Tianjin GC-14C gas-chromatography: FFAP quartz capillary column (0.250 * 30m), carrier gas is a high pure nitrogen, flow 1ml/min, 220 ℃ of temperature of vaporization chamber, column temperature adopts temperature programming, the beginning temperature is 90 ℃, and speed is 3 ℃/min, and outlet temperature is 200 ℃, sample size 1.0 μ l, carry out the qualitative test of organic alcohol according to ordinary method, the color atlas of organic pure standard model such as Fig. 7 a, the retention time under chromatographic condition is as shown in table 7.Organic pure gas chromatogram such as Fig. 7 b and the 7c of crude oil sample before and after the streptococcus zooepidemicus effect.All do not contain three kinds of organic alcohol in the crude oil sample before and after the streptococcus zooepidemicus effect as seen from the figure.
The retention time of the organic pure standard specimen of table 7.
| Ethanol | Propyl alcohol | Butanols | |
| Retention time/min | 1.865 | 2.073 | 2.357 |
(3) evaluation of biogas
Get two test tubes, add 5ml crude oil substratum (in vitro being placed with inverted moral Han Shi tubule) respectively, past wherein test tube inserts 0.5ml streptococcus zooepidemicus CGMCC NO.1988, and another does not connect bacterium as blank.Two test tubes are put into 20-60 ℃ of incubator cultivate 8-72h, result such as Fig. 8, by graph discovery, be connected to the invisible spectro moral Han Shi tubule of streptococcus zooepidemicus aerification, the gas feeding is contained in the clear aqueous solution of saturated calcium oxide, found that calcium oxide aqueous solution becomes muddy, can judge that thus the biogas that streptococcus zooepidemicus produces is a carbonic acid gas.
(4) the hyaluronic evaluation of biological polyoses
1. the hyaluronic preliminary judgement of biological polyoses
Get 5ml streptococcus zooepidemicus (Streptococcus zooepidemicus) CGMCC NO.1988 bacterium liquid and directly be added in the 100ml crude oil, 20-60 ℃, the 120-180r/min shaking table was cultivated 8-72 hour.Whether get the 5ml fermented liquid and add 10ml 95% concentration edible ethanol, observing has throw out to produce.Found that has the cotton shape throw out to occur in the fermented liquid, proving thus has biological polyoses to produce in this fermented liquid, is streptococcus zooepidemicus owing to what use again, is hyaluronic acid so tentatively judge this biological polyoses.
2. infrared absorption is identified
Biological polyoses in the crude oil fermented liquid is separated purification, carry out Infrared spectroscopy then, result such as Fig. 9 b.By (Fig. 9 a) is analyzed, and determines that at last this biological polyoses is exactly a hyaluronic acid with hyaluronic acid standard substance infrared spectra.
The subsequent processes of embodiment 3, streptococcus zooepidemicus CGMCC NO.1988 extraction oil
The subsequent processes of crude oil comprises: dehydration and desalination.
(1) dehydration
Wet oil is defeated must the dehydration to reduce transportation cost before outside the oil field, make crude oil oily water separation, needs carry out breakdown of emulsion to crude oil.Breakdown of emulsion is exactly to be the process of a comparison Unstable Systems with a more stable system transition.At present, Chang Yong breaking method comprises mechanical breakdown of emulsion, electric demulsification and chemical demulsification etc.
The machinery breakdown of emulsion comprises gravity settling, centrifugation, fibre bed filtration, super filtration etc., and mechanical breakdown of emulsion is because its de-emulsification speed is slow, and the facility investment height is so large-scale application is subjected to certain restriction; Electric demulsification is under the high-voltage electric field effect, the liquid pearl is arranged in rows by the direction of electric field, and be elongated to spheroid, the opposite charge of its adjacent both ends has the electrostatic attraction that attracts each other, and this attractive energy causes that the liquid pearl is close mutually, contact and coalescent, reach breakdown of emulsion at last, electric demulsification is because it need consume a large amount of electric energy, and the breakdown of emulsion cost is very high, so brought difficulty to practical application; Chemical demulsification is exactly to utilize the method breakdown of emulsion that adds chemical demulsifier, yet a large amount of uses of chemical demulsifier increase the difficulty of oil product subsequent processes on the one hand, and on the other hand, too much the use of chemical agent brings severe contamination to environment.
Crude oil by streptococcus zooepidemicus CGMCC NO.1988 handles when carrying out crude oil demulsification, need not to add chemical demulsifier, can reach demulsification, result such as Figure 10 as long as crude oil pH is transferred to 9-10.This is because when crude oil system pH is raised to 9-10, and thalline can be dead at once, the crude oil that is in different densities and water under the effect that does not have bacterium with fast separation very, thereby reach the purpose of breakdown of emulsion.Therefore, not only speed is fast when utilizing the crude oil demulsification that streptococcus zooepidemicus CGMCC NO.1988 handles, and cost is very low, helps large-scale application.
(2) desalination
Salt in the crude oil can hydrolysis generates the HCl of severe corrosive, salt also can be on tube wall the formation of deposits salt crust, this not only can reduce thermo-efficiency, increases resistance to flow, even can blocking pipeline; In addition, the salt in the crude oil and also can cause poisoning of catalyst, so crude oil needs to carry out desalting treatment before processing.Separating oil and salt mainly adopt the method for electric desalting at present, and this method not only needs to consume a large amount of electric energy, and desalting effect is not too desirable.
Utilizing streptococcus zooepidemicus CGMCC NO.1988 to carry out intensified oil reduction, is directly bacterial classification to be added in the crude oil system, and does not need additionally to add any nutritive substance in crude oil, and this is different from the method for utilizing microbe oil production in the past.Utilize streptococcus zooepidemicus CGMCC NO.1988 to recover the oil, not only can not increase the salt concn of crude oil, also can the saltiness in the crude oil be reduced greatly because of thalline growth needs in crude oil system consumes most salt in the crude oil, the follow-up desalination operation of crude oil be simplified greatly.So utilize the oily saltiness of streptococcus zooepidemicus CGMCC NO.1988 extraction very low, greatly reduce the crude oil desalting cost, for large-scale application is laid a good foundation.
Description of drawings
Fig. 1 is the molten oily photo of streptococcus zooepidemicus (Streptococcus zooepidemicus) CGMCC NO.1988;
Fig. 2 is streptococcus zooepidemicus (Streptococcus zooepidemicus) CGMCC NO.1988 growth curve;
Fig. 3 is the dispersion and emulsion of streptococcus zooepidemicus (Streptococcus zooepidemicus) CGMCC NO.1988 to crude oil;
Fig. 4 is a crude oil total hydrocarbon color atlas before and after streptococcus zooepidemicus (Streptococcus zooepidemicus) the CGMCC NO.1988 effect;
Fig. 5 is streptococcus zooepidemicus (Streptococcus zooepidemicus) CGMCC NO.1988 effect back viscosity of crude change curve;
Fig. 6 a is an organic acid standard model gas chromatogram;
Fig. 6 b is a crude oil organic acid gas chromatogram before streptococcus zooepidemicus (Streptococcus zooepidemicus) the CGMCC NO.1988 effect;
Fig. 6 c is streptococcus zooepidemicus (Streptococcus zooepidemicus) CGMCC NO.1988 effect back oil sample organic acid gas chromatogram;
Fig. 7 a is organic pure standard model gas chromatogram;
Fig. 7 b is the organic pure gas chromatogram of crude oil before streptococcus zooepidemicus (Streptococcus zooepidemicus) the CGMCC NO.1988 effect;
Fig. 7 c is the organic pure gas chromatogram of streptococcus zooepidemicus (Streptococcus zooepidemicus) CGMCC NO.1988 effect back oil sample;
Fig. 8 is that streptococcus zooepidemicus (Streptococcus zooepidemicus) CGMCC NO.1988 effect back produces biogas CO
2
Fig. 9 a is biological polyoses hyaluronic acid standard substance infrared spectrograms;
Fig. 9 b is streptococcus zooepidemicus (Streptococcus zooepidemicus) CGMCC NO.1988 effect back oil sample biological polyoses infrared spectrogram;
Figure 10. behind streptococcus zooepidemicus (Streptococcus zooepidemicus) the CGMCC NO.1988 extraction oil breakdown of emulsion.
Claims (3)
1, streptococcus zooepidemicus (Streptococcus zooepidemicus) CGMCC NO.1988 is in the petroleum production engineering Application for Field.
2, streptococcus zooepidemicus (Streptococcus zooepidemicus) CGMCC NO.1988 is in the application of oil transport field.
3, streptococcus zooepidemicus (Streptococcus zooepidemicus) CGMCC NO.1988.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101538459B (en) * | 2009-04-30 | 2011-08-03 | 中国石油天然气股份有限公司 | Oil well pollution-free paraffin removing and viscosity reducing agent |
| CN104059865A (en) * | 2014-06-11 | 2014-09-24 | 滨州安华生物工程有限公司 | Streptococcus zooepidemicus and production process for preparing hyaluronic acid by using same |
| CN111909860A (en) * | 2020-07-30 | 2020-11-10 | 克拉玛依市新奥达石油技术服务有限公司 | Bacterial strain for producing biological emulsifier and biological emulsifier produced by using bacterial strain |
| CN112225324A (en) * | 2020-09-16 | 2021-01-15 | 山东大学 | Screening method of nitrate reducing bacteria for demulsification of strong-emulsification ASP flooding produced water |
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2007
- 2007-04-03 CN CNB2007100906957A patent/CN100503813C/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101538459B (en) * | 2009-04-30 | 2011-08-03 | 中国石油天然气股份有限公司 | Oil well pollution-free paraffin removing and viscosity reducing agent |
| CN104059865A (en) * | 2014-06-11 | 2014-09-24 | 滨州安华生物工程有限公司 | Streptococcus zooepidemicus and production process for preparing hyaluronic acid by using same |
| CN104059865B (en) * | 2014-06-11 | 2016-08-24 | 山东安华生物医药股份有限公司 | A kind of streptococcus zooepidemicus and prepare the production technology of hyaluronic acid with it |
| CN111909860A (en) * | 2020-07-30 | 2020-11-10 | 克拉玛依市新奥达石油技术服务有限公司 | Bacterial strain for producing biological emulsifier and biological emulsifier produced by using bacterial strain |
| CN111909860B (en) * | 2020-07-30 | 2022-06-21 | 克拉玛依市新奥达石油技术服务有限公司 | Bacterial strain for producing biological emulsifier and biological emulsifier produced by using bacterial strain |
| CN112225324A (en) * | 2020-09-16 | 2021-01-15 | 山东大学 | Screening method of nitrate reducing bacteria for demulsification of strong-emulsification ASP flooding produced water |
| CN112225324B (en) * | 2020-09-16 | 2021-09-17 | 山东大学 | Screening method of nitrate reducing bacteria for demulsification of strong-emulsification ASP flooding produced water |
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| CN100503813C (en) | 2009-06-24 |
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