CN1285827C - Microbe oil-displacement method and microbe trielement compound oil-displacement agent - Google Patents

Abstract

The present invention discloses an oil displacement method which orderly comprises the following procedures: at least one of bacillus cereus HPCGMCC No. 1141 and brevibacillus brevis HTCGMCC No. 1142 is cultivated in a culture medium which takes crude oil as a carbon source to obtain fermentation liquor; the obtained fermentation liquor is injected in an oil layer for 3 to 5 days; oil is displaced by a ternary compounding system. After the microorganism fermentation liquor obtained by HP and HT acts on crude oil, the physical properties of the crude oil are improved, the interfacial tension of the acted crude oil and the existing ternary formula is lower than that of no acted crude oil and the existing ternary formula, a small amount of alkylbenzene sulfonate surfactant is added to the microorganism fermentation liquor diluted by 2 to 12 times, the interfacial tension can achieve 10<-3>mN/m, the cost is greatly reduced, and the present invention has good stability.

Description

A kind of microorganism-ternary composite oil-displacing method and a kind of microorganism-ternary composite driving finish

Technical field

The present invention relates to oil exploitation technology, thereby particularly using microbe acts on the method that crude oil improves oil displacement efficiency, and the oil displacement agent of using this microorganism and three-component compound system assembly.

Background technology

The world today is carrying out one with bioengineering and be applied as the new technology revolution of sign, and biological develop energy technology becomes economic potential maximum in the bioengineering, is hopeful one of field with future most, analysis of modernization instrument and means also make the microorganism science enter a brand-new developing period, and these factors have promoted microbial enhanced oil recovery Study on Technology and application.

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 use the chemical method crude oil extraction, and the ASP ternary composite oil-displacing system of forming as alkali, surfactant, polymer (Mu Jianhai, prune is helped .[J]. chemical science and technology market .2000,23 (7): 17).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.The technology that this use chemistry or biologic product improve recovery ratio is called as tertiary oil recovery technology, also claims intensified oil reduction (EOR, Enhanced OilRecovery) technology abroad.For they are made a distinction, improve recovery efficiency technique with microorganism and also can be called four oil recovery techniques or microbial enhanced oil recovery (MEOR, Microbial Enhanced Oil Recovery) technology, it is meant and utilizes the useful metabolism product of micro-organisms or utilize microorganism can decompose the performance of hydrocarbon, thus the technology of raising 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 old filed of height.The microbial enhanced oil recovery technology mainly comprise two classes (Li Yugeng. petroleum microbiology .[M]. Shanghai: the .1996 of publishing house of Shanghai Communications University): a class is to utilize microniological proudcts such as biopolymer and biosurfactant to carry out the displacement of reservoir oil as the oil field with chemical agent, be called microorganism ground top fermentation and improve recovery ratio technology, be the bioprocess technology method, this technology has become ripe at home and abroad at present; Another kind of is to utilize microorganism and metabolite 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 of degraded oil microorganism 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, Beck as Pennsylvania crude oil association has reported the experiment that discharges crude oil with sulfate reducing bacteria, studies show that the good result (nineteen forty-seven) that sulfate reducing bacteria occurs in the bacterium oil decomposes, this experiment was drawn striking by the general Dege of strategic point afterwards and reyn further confirms (1954).Larry dimension thunder (nineteen fifty-five) has observed pseudomonas aeruginosa, mycobacterium, bacillus subtilis, lipolytic enzyme read the surface tension in cultures such as glycerine such as coccus, and surface tension descends obviously, but utilizes hydro carbons and bad.Being necessary to seek more effectively, bacterial classification is improved oil recovery efficiency with the use more efficient methods.

Summary of the invention

The objective of the invention is in the method that provides microorganism that a kind of utilization can degraded oil to carry out the displacement of reservoir oil.

Flooding method of the present invention may further comprise the steps in proper order:

1) with Bacillus cercus (Bacillus cereus) HP, CGMCC № .1141 and short bacillus (Brevibacillus brevis) HT, at least one strain is cultivated in the culture medium that with crude oil is carbon source and is obtained zymotic fluid among the CGMCC № .1142;

2) zymotic fluid that obtains was injected oil reservoir 3 to 7 days;

3) with the displacement of reservoir oil of ternary built system.

Above-mentioned flooding method, culture medium A is K in the described step 1) ₂HPO ₄0.1%, NaH ₂PO ₄0.2%, (NH ₄) ₂SO ₄0.2%, CaCl ₂2H ₂O 0.001%, FeSO ₄7H ₂O 0.001%, yeast 0.1%,, PH7.0-7.2, carbon source is a 10-20% crude oil; Condition of culture is 45 ℃ of temperature, shaking speed 120rpm, incubation time 7 days;

Another kind of culture medium B is KH ₂PO ₄: 0.1-1%, Na ₂HPO ₄: 0.05-1%, NH ₄Cl:0.05-1%, NaCl:0.1-1.0%, MgSO ₄7H2O:0.01-0.5%, KCl:0.05-0.5%, FeCl ₂: 0.001-0.01%, CaCl ₂: 0.001-0.01%, MnCl ₂: 0.001-0.01%, CuCl ₂: 0.001-0.01%, ZnCl ₂: 0.001-0.01%, yeast soaks powder: 0.01-0.2%, beef extract: 0.01-0.2%, peptone: 0.01-0.2%, atoleine 0.5-2% or crude oil 1-20%, Tween80:0.01-0.2%, Tween95:0.01-0.2%, carboxylate: 0.005-0.1%, pH:6.8-7.5,121 ℃ of sterilization 15-20Min.

In the above-mentioned flooding method, step 2) its concentration of zymotic fluid of the injection oil reservoir in is 1～5wt%, wherein contains 10 ⁷-10 ⁸Individual cell/ml.

In the above-mentioned flooding method, in the described step 3) in the ternary built system each component be: the zymotic fluid 8wt%-50wt% that step 1) obtains, table live agent 0.04wt%, alkali 1.0wt% and polymer 0.25wt%.

In the above-mentioned flooding method, in the step 3) in the ternary built system each component be: table live agent 0.2wt%, alkali 1.0wt% and polymer 0.25wt%.

Wherein, microorganism that can degraded oil is Bacillus cercus (Bacillus cereus) HP or short bacillus (Brevibacillus brevis) HT, wherein Bacillus cercus (Bacillus cereus) HP was preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center (being called for short CGMCC) on 04 08th, 2004, and preserving number is CGMCC № .1141; Short bacillus (Brevibacillus brevis) HT has been preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center (being called for short CGMCC) on 04 08th, 2004, preserving number is CGMCC № .1142.

Another purpose of the present invention is to provide a kind of ternary composite driving finish that utilizes mentioned microorganism that can effectively improve oil recovery.

Microorganism provided by the invention-ternary composite driving finish comprises following component:

Microbial fermentation solution 8%-50wt%,

Table agent 0.04wt% alive,

Alkali 1.0wt% and

Polymer 0.25wt%;

Wherein, microbial fermentation solution is Bacillus cercus (Bacillus cereus) HP, CGMCC № .1141, or short bacillus (Brevibacillus brevis) HT, CGMCC № .1142 bacterial classification is cultivated in the culture medium that with crude oil is carbon source and is obtained zymotic fluid.

Experimental results show that, Bacillus cercus (Bacillus cereus) HP and short bacillus (Brevibacillusbrevis) HT all can improve the character of crude oil effectively, acid value for crude oil improves more than 8 times, light component increases, the content of wax contains glue reduced rate 30-50%, rheological characteristic of crude oil improves, and interfacial tension reduces between profit, and content of material such as organic acid increases in the zymotic fluid; Its metabolism crude oil approach is time terminal oxidation, has produced a kind of novel lipid surfactant.

Experiment is proof also, after utilizing the microbial fermentation solution effect crude oil that HP, HT obtain, crude oil property improves, effect back crude oil and existing ternary prescription interfacial tension effect crude oil reduce, doubly (concentration 50%～8wt%) microbial fermentation solution adds a spot of alkylbenzenesulfonate table agent S1 (0.01wt%-0.04wt%) alive to dilution 2-12, and interfacial tension can reach 10 ^-3MN/m greatly reduces cost, and has stability preferably.

The laboratory core oil displacement experiment shows, under the situation that adds a small amount of sulfonate table agent alive (0.04%), 2 times (50wt%) of microbial fermentation solution dilution that utilizes HP, HT to obtain, the indoor natural core displacement of reservoir oil improves the recovery ratio amplitude on average 20.25% after the zymotic fluid effect, suitable with existing ternary prescription, but can save the consumption of surfactant in the former ternary prescription; Annotate zymotic fluid earlier,, improve the recovery ratio amplitude, increase about 9% than independent three-component compound system displacement of reservoir oil amplitude on average 29% with the displacement of existing ternary prescription.

Description of drawings

Fig. 1 a is a HP effect back crude oil total hydrocarbon chromatogram

Fig. 1 b is a HT effect back crude oil total hydrocarbon chromatogram

Fig. 2 is original rheological characteristic change curve before and after bacterial classification HP, the HT effect

Fig. 3 a be blank oil sample nonhydrocarbon micro--the infrared analysis collection of illustrative plates

Fig. 3 b be HP effect back oil sample nonhydrocarbon micro--the infrared analysis collection of illustrative plates

Fig. 3 c be HT effect back oil sample nonhydrocarbon micro--the infrared analysis collection of illustrative plates

Fig. 4 is an alkane microbiological oxidation general way

Fig. 5 is an alkane time terminal oxidative pathway

Fig. 6 a is the chromatogram of organic acid mixing standard specimen

Fig. 6 b is an organic acid chromatogram in the HT sample

Fig. 6 c is an organic acid chromatogram in the HP sample

Fig. 7 a is the mass spectrogram of acetate

Fig. 7 b is the mass spectrogram of propionic acid

Fig. 7 c is the mass spectrogram of butyric acid

Fig. 7 d is the mass spectrogram of isovaleric acid

Fig. 8 a is the chromatogram of organic pure standard specimen

Fig. 8 b is the organic pure chromatogram of HP sample

Fig. 8 c is the organic pure chromatogram of HT sample

Fig. 9 a is the infrared spectrogram of the B1 material that extracts in the HT sample

Fig. 9 b is the infrared spectrum of the B2 material that extracts in the HT sample

Fig. 9 c is the infrared spectrum of the C material that extracts in the HT sample

Fig. 9 d is the infrared spectrogram of the B material that extracts in the HP sample

Fig. 9 e is the infrared spectrogram of the C material that extracts in the HP sample

Figure 10 is the correlation curve figure of the interfacial tension of microbial fermentation solution one ternary system

Figure 11 is an interfacial tension between ternary system and effect front and back crude oil

Figure 12 a is an extraordinarily interfacial tension between alkali (0.8) profit of microbial fermentation solution dilution 4

Figure 12 b is an extraordinarily interfacial tension between alkali (1.0) profit of microbial fermentation solution dilution 4

Figure 12 c is an extraordinarily interfacial tension between alkali (0.6) profit of microbial fermentation solution dilution 4

Figure 13 a is that in the time of 30 minutes the active region for zymotic fluid A and A effect crude oil

Figure 13 b is that in the time of 120 minutes the active region for zymotic fluid A and A effect crude oil

Figure 13 c is that in the time of 30 minutes the active region for zymotic fluid B and B effect crude oil

Figure 13 d is that in the time of 120 minutes the active region for zymotic fluid B and B effect crude oil

The specific embodiment

" % " that mentions in technical solution of the present invention and embodiment if no special instructions, is mass percentage concentration.

The screening of embodiment 1, bacterial classification

1, Bacillus cercus (Bacillus cereus) HP takes a sample and enrichment culture with being divided into separating of short bacillus (Brevibacillusbrevis) HT, screening and two steps of purifying, and concrete grammar is as follows:

(1) sampling and enrichment culture

Get behind the profit sample of first oil recovery factory of Daqing, Heilongjiang Province enrichment culture immediately, required bacterial classification is quantitatively preponderated, suppress unwanted growth.What enriched medium was used is minimal medium: fluid nutrient medium A:K ₂HPO ₄: 0.1-1%, NaH ₂PO ₄: 0.1-0.5%, (NH ₄) ₂SO ₄: 0.05-0.2%, MgSO ₄7H2O:0.01-0.5%, FeCl ₂: 0.001-0.01%, CaCl ₂: 0.001-0.01%, yeast soaks powder: 0.02-0.2%, crude oil 0.5-20%.121 ℃ of pH:6.8-7.5, sterilization 15-20min.45 ℃ of 120rpm shaking tables were cultivated 5-7 days.

(2) screening of bacterial classification and purifying

Two kinds of selective mediums of screening usefulness are crude oil inorganic salts solid medium and blood plate culture medium.

A. the screening on the crude oil inorganic salts solid medium

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.The described enrichment inorganic salts of preparation process under aseptic condition (1) solid medium flat board, crude oil after the dilution is poured into (addition of crude oil is 1ml crude oil/L culture medium) on the inorganic salts solid medium flat board for preparing, after treating that crude oil is tiled in the inorganic salts planar surface equably and solidifies, the enrichment culture liquid in the step (1) is uniformly coated on the flat board.Cultivated 3～5 days for 45 ℃, can find out significantly that the crude oil in long bacterium place is utilized, the former oil reservoir attenuation on the inorganic salts flat board forms transparent circle.Selection can form the bacterial classification of transparent circle, waits to be further purified.

B. the screening of blood plate cultivation

Enrichment culture liquid in the step (1) is uniformly coated on the blood plate.Cultivated 1-2 days for 45 ℃, show that the periphery of bacterial colonies after the cultivation forms tangible haemolysis transparent circle, select such bacterium colony and wait to be further purified.Since biosurfactant have can haemolysis characteristic, single bacterium colony of haemolysis promptly is the bacterial classification that produces biosurfactant.The composition of blood plate culture medium: beef extract 0.3%, peptone 1%, yeast extract 0.01%, sodium chloride 0.5%, agar 2%, sheep blood 5%.

C. will dye down by single colony microscope that above-mentioned selective medium obtains and observe, and, utilize above-mentioned culture medium to separate again, up to obtaining pure bacterium colony if impure.In order to obtain anaerobism and facultative bacterial classification, the bacterial classification that obtains by above-mentioned two kinds of selective mediums in the anaerobism work station 45 ℃ cultivated 5 days, acquisition is a strain bacterium HP and a HT of sole carbon source with crude oil, and this two strains bacterium has good product surfactant, produces sour aerogenesis, improves characteristics such as crude oil property.

Can select for use following two kinds of methods that two bacterial strains are carried out long-term culture presevation: 1. low-temperature preservation: to get the bacterial classification that is in exponential phase, make protective agent-70 ℃ of preservations with 15% aseptic glycerine; 2. freeze drying prepares dry powder: centrifugal zymotic fluid obtains thalline, makes protective agent with skim milk, and dry powder is made in freeze drying.

2, condition of culture

Utilize Orthogonal Experiment and Design, determined the prescription of culture medium:

Culture medium A is K ₂HPO ₄0.1%, NaH ₂PO ₄0.2%, (NH ₄) ₂SO ₄0.2%, CaCl ₂2H ₂O 0.001%, FeSO ₄7H ₂O 0.001%, yeast 0.1%, and, PH 7.0-7.2.Carbon source is a 10-20% crude oil; Condition of culture is: cultivation temperature 30-50 ℃, and shaking speed 120rpm, incubation time 5-7 days.

Another kind of culture medium B is KH ₂PO ₄: 0.1-1%, Na ₂HPO ₄: 0.05-1%, NH ₄Cl:0.05-1%, NaCl:0.1-1.0%, MgSO ₄7H2O:0.01-0.5%, KCl:0.05-0.5%, FeCl ₂: 0.001-0.01%, CaCl ₂: 0.001-0.01%, MnCl ₂: 0.001-0.01%, CuCl ₂: 0.001-0.01%, ZnCl ₂: 0.001-0.01%, yeast soaks powder: 0.01-0.2%, beef extract: 0.01-0.2%, peptone: 0.01-0.2%, atoleine 0.5-2% or crude oil 1-20%, Tween80:0.01-0.2%, Tween95:0.01-0.2%, carboxylate: 0.005-0.1%, pH:6.8-7.5,121 ℃ of sterilization 15-20Min; Condition of culture is: cultivation temperature 30-50 ℃, and shaking speed 120rpm, incubation time 1-2 days.

Embodiment 2, bacterial classification are to the effect of crude oil

1, dispersion and emulsion effect experiment

Bacillus cercus (Bacillus cereus) HP bacterial classification and short bacillus (Brevibacillusbrevis) HT bacterial classification are at K ₂HPO ₄0.1%, NaH ₂PO ₄0.2%, (NH ₄) ₂SO ₄0.2%, CaCl ₂2H ₂O 0.001%, FeSO ₄7H ₂O 0.001%, yeast 0.1%, and, PH 7.0-7.2.Carbon source 10% crude oil.45 ℃, the result that the 120rpm shaking table was cultivated after 5 days shows: compare with contrast (not adding bacterium), the color of zymotic fluid is obviously deepened, and effect back crude oil has the advantages that not hang bottle.Because microorganism is the sole carbon source growth with crude oil, changed the character of crude oil, make the component of crude oil that variation take place.Crude oil after the fermentation is examined under a microscope, and finds to have in the crude oil bacterium liquid to exist.Bacterium exists on the oil-water interfaces, and is carbon source with oil, conforms to produce displacement of reservoir oil material, thereby plays emulsification, wetting, the effect that disperses crude oil.

2, crude oil total hydrocarbon chromatogram before and after the bacterial classification effect

Get the crude oil of above-mentioned fermentation after 5 days, do the total hydrocarbon analysis of crude oil according to a conventional method.The result shows optionally some high carbon number alkane in the degrading crude oil of two bacterial strains shown in table 1 and Fig. 1 a and Fig. 1 b, long chain hydrocarbon content reduces relatively, and short hydrocarbon or low chain hydrocarbon content increase relatively, and the light components of crude oil increases; In addition, ∑ C ₂₁/ ∑ C ₂₂And C ₂₁+ C ₂₂/ C ₂₈+ C ₂₉Be the parameter of describing oil-gas migration, ∑ C so ₂₁/ ∑ C ₂₂And C ₂₁+ C ₂₂/ C ₂₈+ C ₂₉Ratio increase, the direction of expression oil migration, along with macromolecular compound content reduces relatively, the light component compounds content increases relatively.Fig. 1 a and Fig. 1 b show because the thermal degradation of high-molecular weight hydrocarbon and shifting to the low carbon number scope, and chromatogram gradually becomes preceding high cutting edge of a knife or a sword type, and normal alkane carbon number distribution curve moves to light component direction.The flowability of crude oil improves.

The saturated hydrocarbons chromatogram testing result of the crude oil after the table 1. bacterial strain HP fermentation

Sample	Carbon number range	Main peak carbon	∑C21 ∑C22	C21+C22 C28+C29
					Blank	nc9-nc35	nc23	0.67	1.66
HP	nc9-nc34	nc16	1.30	1.82
					HT	nc9-nc35	nc23	1.62	1.89
The HT+HP Mixed Microbes	nc9-nc35	nc18	1.24	2.06

3, rheological characteristic of crude oil situation of change

Two strain bacterium are at K ₂HPO ₄0.1%, NaH ₂PO ₄0.2%, (NH ₄) ₂SO ₄0.2%, CaCl ₂2H ₂O 0.001%, FeSO ₄7H ₂O 0.001%, yeast 0.1%, PH 7.0-7.2.Carbon source is 10% crude oil.45 ℃, after the 120rpm shaking table is cultivated 5 days, analyze microbial action front and back rheological characteristic of crude oil according to a conventional method respectively, the result shows that effect back rheological characteristic of crude oil obviously improves as shown in Figure 2, viscosity reduces.When rotor revolution number was 6l/s, viscosity of crude was 101mPas before the effect, and bacterial strain HT effect back viscosity of crude is reduced to 56.9mPas, viscosity of crude has reduced by 43.7%, bacterial strain HP effect back viscosity of crude is reduced to 43.4mPas, and viscosity of crude has reduced by 57.03%, and viscosity of crude declines to a great extent.

4, the crude oil content of wax, contain glue and change

After having analyzed crude oil alkane distribution situation, in order further to inquire into the situation of change of the bacterial strain effect front and back crude oil content of wax, glue, do the crude oil content of wax according to a conventional method, contained glue variation experiment, the result is as shown in table 2, the content that shows the wax of crude oil before and after the bacterial classification effect and colloid all has variation in various degree, and wax content in crude oil descends after the bacterial strain effect.

The change list of crude oil wax, glue before and after the effect of table 2. bacterial classification

The sample title	Content of wax %	Contain glue %
			Blank crude oil	20.2	19.8
HT	12.6	15.1
			HP	14.3	14.7

5, the variation of aromatic hydrocarbons in the crude oil

Get the crude oil of fermentation after 5 days and carry out GC-MS according to a conventional method, the relative amount of the Sino-Philippines series of analyse crude oil aromatic hydrocarbons, the result is as shown in table 3, shows optionally degraded different component in the luxuriant and rich with fragrance series of bacterial strain, causes MPI, DPI index variation.Luxuriant and rich with fragrance series in the crude oil is commonly considered as the cracking from steroid terpene compound.Because alkyl position difference on phenanthrene ring, stability is also variant, generally be in the methyl of β position, more stable as 3-methyl and 2-methylphenanthrene, at the 1-of α position methyl, 4-methylphenanthrene and to be in the 9-methylphenanthrene of meta more active, dimethylphenanthrene also has similarity rules, to β β type, stability increases successively from α α type, α β type, after this bacterial strain effect, the MPI of crude oil and DPI index all improve, and illustrate that the comparatively active component of chemical property in the luxuriant and rich with fragrance series is easily by this strains for degrading.

Aromatic hydrocarbons GC-MS analysis result in the crude oil before and after the table 3. bacterial strain HP effect

Sequence number	The sample title	Luxuriant and rich with fragrance serial content (%)	MPI	DPI
						1 2 3	Blank oily HT oil HP oil	7.26 7.21 6.54	0.67 0.72 0.89	0.87 0.91 1.02
Remarks: MPI=1.5 * (2-methylphenanthrene+3-methylphenanthrene)/(phenanthrene+1-methylphenanthrene+9-methylphenanthrene) DPI=4 * (dimethylphenanthrene 1.+dimethylphenanthrene 2.+dimethylphenanthrene 3.+dimethylphenanthrene 4.)/(phenanthrene+dimethylphenanthrene 5.+dimethylphenanthrene 6.+dimethylphenanthrene 7.)

6, the variation of nonhydrocarbon in the crude oil

Get the crude oil of fermentation after 5 days and adopt micro--infrared method to analyze before and after the two bacterial strain effects variation of non-dydrocarbon constituents and structure in the crude oil, the result is shown in Fig. 3 a and Fig. 3 b (HP oil sample), Fig. 3 c (HT oil sample), and Fig. 3 a shows 2924cm ^-1And 2853cm ^-1The absworption peak difference represent methylidene at place and the asymmetric and symmetrical stretching vibration of C-H of methylene, methylene is dominant, because the C-H stretching vibration peak of general methyl is at 2960cm ^-1And 2870cm ^-1The place is covered by methylene peak, adds 722cm ^-1The peak crude oil nonhydrocarbon is described based on long carbon saturated chain, and the side chain degree is little for the absworption peak of methylene in the long carbochain (n＞4); 1461cm ^-1And 1376cm ^-1The C-H flexural vibrations peak of place's methyl, methylene is the circumstantial evidence of above deduction.1601cm ^-1The absworption peak at place may be that the two key stretching vibrations of C=N, C=C cause, but according to fingerprint region 910cm ^-1Following not obvious infrared absorption peak (relevant with the position of substitution of phenyl ring) can be inferred and be contained a certain amount of aromatic hydrocarbons composition in the crude oil nonhydrocarbon.1650-1900cm ^-1Absworption peak in the scope mainly shows as carbonyls, as the C-O stretching vibration of aldehyde, ketone, acid, ester, acid anhydrides and acid amides etc., from infrared spectrum, a little less than the absorption intensity at the interior peak of this scope, and becoming broadband shape, may be because contain the mixing cpd of a small amount of band carbonyl in the crude oil nonhydrocarbon.Fig. 3 b show after the effect of HP bacterial strain with microbial action before to compare the main distinction be at 3303cm ^-1New infrared absorption peak appears in the place, Fig. 3 c show after the effect of HT bacterial strain with microbial action before to compare the main distinction be at 2463cm ^-1New infrared absorption peak appears in the place, and peak shape is wide and blunt, and this produces owing to hydroxyl forms hydrogen bond in intermolecular or molecule, but at 1603cm ^-1The place does not have tangible infrared absorption peak, illustrates that hydrogen bond is not to cause because contain water in the sample.According to the skew rule of peak shape feature and chemical shift, there is a certain amount of carboxylic acid to generate in the luxuriant and rich with fragrance hydrocarbon of crude oil after the effect of deduction bacterial strain, the interior strong association owing to carbonyl and hydroxyl of carboxylic acid can be from 3300cm according to the different infrared absorption peaks of carboxylic acid content ^-1Be extended to 2500cm ^-1Below.

For further confirmation, measured the acid number of bacterial strain effect front and back crude oil nonhydrocarbons, the result is as shown in table 4, show after the bacterial strain effect, the acid number of crude oil nonhydrocarbon is increased to original 8 times and 14 times, illustrate that bacterial strain in the degradation process to the crude oil different component biooxidation reactions has taken place, generated high-carbon organic acid, improved the acid number of crude oil.

Crude oil nonhydrocarbon acid number analysis result before and after the effect of table 4. bacterial strain

Sequence number	The sample title	Nonhydrocarbon acid number (%)	Acid number increases (doubly)
				1 2 3	Blank oily HP oil HT oil	0.1037 0.8362 1.4799	8.06 14.27

7, saturated hydrocarbons biodegradation Analysis on Mechanism in the crude oil

From above research as can be seen, bacterial strain to the degraded of crude oil to be degraded to main path.Most of microbe can be summarized with Fig. 4 the main oxidative pathway of alkane, at first is alkane to be carried out terminal oxidation (or beta oxidation) generate aliphatic acid, press beta oxidation approach degraded alkane again, and association acyl group-CoA participates in the metabolic activity of microorganism.Usually, for a specific alkane, microbial degradation product aliphatic acid is one to differ the mixed acid of two carbon each other, and just degradation condition and carbon number is different, and the carbon number distribution of mixed acid is different.Bacterial strain HP, HT show that to the constituent analysis result of oil degradation the carbon number that mixes fat acid acid concentrates on C ₂-C ₂₀In the scope.This explanation bacterial strain may be since a comparatively special approach to the biological oxidation of crude oil---inferior terminal oxidation (subterminaloxidation) (as Fig. 5).Crude oil medium high carbon chain saturated hydrocarbons is the aliphatic acid of carbochain in time terminal oxidation generates two kinds at first, carries out terminal oxidation and beta oxidation again.

The producing and analyzing of embodiment 3, bacterial strain fermentation liquor

1, producing of zymotic fluid: will in embodiment 1, cultivate 5 days among the culture medium selected A with Bacillus cercus (Bacillus cereus) HP and short bacillus (Brevibacillus brevis) the HT bacterial classification of the screening of embodiment 1 method, or in culture medium B, cultivate after 2 days, 3500rpm is centrifugal with refrigerated centrifuge, remove upper strata crude oil, collect thalline and nutrient solution and obtain zymotic fluid.

2, the analysis of zymotic fluid

(1) organic acid The qualitative analysis

Utilize 6890N/5973N gas chromatograph-mass spectrometer (U.S. Agilent company), (0.25mm * 30m), carrier gas is high-purity helium to the FFAP quartz capillary column, flow 1ml/min, 250 ℃ of temperature of vaporization chamber, column temperature adopts temperature programming, and the beginning temperature is 160 ℃, and speed is 2 ℃/min, final temperature is 180 ℃, sample size 0.5 μ l. carries out the organic acid qualitative determination according to conventional method, the chromatogram of organic acid standard specimen such as Fig. 6 a, and the retention time under chromatographic condition is as shown in table 5.

The organic acid gas chromatogram of HP sample such as Fig. 6 c, the organic acid gas chromatogram of HT sample such as Fig. 6 b, contrast through Mass Spectrometer Method with the organic acid standard specimen, containing four kinds of organic acids in the sample is acetate, propionic acid, butyric acid, isovaleric acid, and organic acid Mass Spectrometer Method figure is shown in Fig. 7 a, 7b, 7c and 7d.

The retention time of table 5. organic acid standard specimen

	Acetate	Propionic acid	Butyric acid	Isovaleric acid
					Retention time (min)	2.16	2.50	2.99	3.26

(2) organic acid quantitative analysis results

The test of water sample distilling effect

This extracting method (is measured pretreated sample 100ml, is placed distilling flask, add the acidifying of 7ml phosphoric acid, distillation.When distilling flask is left small volume of solution, cold slightly, respectively add 20ml distilled water more at twice and continue distillation.With in the NaOH solution and distillate, PH is 8～9 with acidometer control.After being concentrated into about 5ml with Rotary Evaporators, sample is changed in the 10ml volumetric flask, with hcl acidifying to PH≤3, constant volume, organic acid to be measured) can extract various organic acids fully, done the distilling effect test.Promptly respectively get five portions of HP zymotic fluids of 100ml, handle through said method and measure, the result is as shown in table 6:

The test of table 6. organic acid distilling effect

Organic acid	Standard deviation	(five times average: mg/L) for measured value		The coefficient of variation (%)
					The second third fourth isoamyl	Acid acid acid acid	24.35 20.41 3.082 1.269	1.02 0.73 0.16 0.37	4.2 3.6 5.3 1.6

Table 6 shows: five measurement result reappearances of same sample are better, illustrate to adopt the method for distillation fully the low molecular weight organic acid in the water sample to be steamed.

With HP, HT bacterial classification at K ₂HPO ₄0.1%, NaH ₂PO ₄0.2%, (NH ₄) ₂SO ₄0.2%, CaCl ₂2H ₂O 0.001%, FeSO ₄7H ₂O 0.001%, yeast 0.1%, and, PH 7.0-7.2.Carbon source is 10% crude oil.45 ℃, nutrient solution after the 120rpm shaking table is cultivated 5 days is made three parallel samples respectively respectively, press 6890N/5973N gas chromatograph-mass spectrometer (U.S. Agilent company), the FFAP quartz capillary column (0.25mm * 30m), carrier gas is high-purity helium, flow 1ml/min, 250 ℃ of temperature of vaporization chamber, column temperature adopts temperature programming, the beginning temperature is 160 ℃, speed is 2 ℃/min, and final temperature is 180 ℃, and sample size 0.5 μ l measures respectively, obtain organic acid concentration average, result such as table 7, shown in the table 8, show HP, acetic acid content is bigger in the HT sample, based on acetate.

Design formulas:

(ρ is a standard specimen density to standard specimen concentration (mg/L)=20/50 * ρ * 1000=400 ρ, g/ml)

Organic acid quantitative analysis results in the table 7.HP sample

	Concentration (mg/L)	Standard deviation	The coefficient of variation (%)	Molar concentration (mmol/L)
					Acetate propionic acid butyric acid isovaleric acid total acid content	23.22 21.12 2.932 7.201	1.49 1.18 0.12 0.47 0.7763	6.4 5.6 4.1 3.6 mmol/L	0.3870 0.2854 0.0333 0.07060

Organic acid quantitative analysis results in the table 8.HT sample

	Concentration (mg/L)	Standard deviation	The coefficient of variation (%)	Molar concentration (mmol/L)
					Acetate propionic acid butyric acid isovaleric acid total acid content	574.0 8.318 14.68 7.131	45.3 0.44 1.15 0.83 9.9158	7.9 5.3 7.8 6.1 mmol/L	9.5667 0.1124 0.1668 0.0699

3, the organic pure qualitative and quantitative analysis of zymotic fluid

Utilize 6890N/5973N gas chromatograph-mass spectrometer (U.S. Agilent company), the FFAP quartz capillary column (0.25mm * 30m), carrier gas is high-purity helium, flow 1ml/min, 200 ℃ of temperature of vaporization chamber, column temperature adopts temperature programming, the beginning temperature is 80 ℃, kept 1 minute, speed is 4 ℃/min, 110 ℃ of final temperatures, sample size 1.0 μ l. carry out the qualitative determination of organic alcohol according to conventional method, the chromatogram of organic pure standard specimen shows under this chromatographic condition that shown in Fig. 8 a (former Figure 24) four kinds of organic pure separating effects are fine; Retention time under chromatographic condition is shown in table 9a, 9b.

The organic pure gas chromatogram of HP sample shown in Fig. 8 b, the organic pure gas chromatogram of HT sample such as Fig. 8 c.Through Mass Spectrometer Method, only contain ethanol in the HT sample, do not detect organic alcohol in the HP sample.

The retention time of the organic pure standard specimen of table 9a.

	Ethanol	Propyl alcohol	Butanols	Amylalcohol
					Retention time (min)	1.93	2.47	3.51	5.00

Ethanol quantitative result in the HT sample is as table 9b.Wherein 20 μ l ethanol constant volumes are in the 50ml volumetric flask, and concentration is 315.7m g/L, the same organic acid of quantitative analysis computational methods.

The quantitative analysis results of table 9b.HT sample ethanol

	Concentration (mg/L)	Standard deviation	The coefficient of variation (%)	Molar concentration (mmol/L)
					Ethanol	759.9	28.5	3.8	16.52

4, the evaluation of biosurfactant

(1) Preliminary Identification of biosurfactant

1. the evaluation of lipopeptid type biological surfactant

Get each 50ml of HP, HT and HP+HT, transfer pH value to 2 with concentrated hydrochloric acid, 4 ℃ of standing over night, whether observe has precipitation to produce.The result shows that three kinds of zymotic fluids all do not have precipitation and produce, and proves thus in three kinds of zymotic fluids and does not contain the lipopeptid type biological surfactant.

2. UV absorption is identified

Biosurfactant after purifying has according to a conventional method been done determination of uv absorption, and they all do not have absorption in ultraviolet range.Can tentatively judge and not contain aromatic compound.

(2) elementary analysis of biosurfactant

The biosurfactant of purifying and obtaining from HP and HT zymotic fluid carries out C, H, N Determination on content to it respectively according to a conventional method, and data are as shown in table 10:

Table 10 shows the content of these five kinds of biosurfactant N all below 3.5%, and the content of N is all more than 10% in the amino acid of reporting on the document, and as seen, these biosurfactants are not amino acid and lipopeptid class.And detect with ninhydrin solution, amino acid whose characteristic reaction do not occur, further determined above-mentioned conclusion.

The results of elemental analyses of table 10. biosurfactant

	C(％)	H(％)	N(％)
				Extract among the C material HT that extracts among the B material HP that extracts among the HP to extract among the B2 material HT among the B1 material HT and extract the C material	73.26 73.64 73.06 77.30 66.33	12.23 10.82 11.29 15.53 8.71	3.01 3.15 3.50 2.93 2.38

(3) infrared spectrum analysis of biosurfactant

The infrared spectrum of the B material that HP, HT sample obtain after purifying shown in Fig. 9 a/9b/9c, shows 2924--2853cm respectively ^-1, 1456cm ^-1Be the C-H vibration peak of methyl, methylene, do not have the characteristic peak of other biosurfactant.

The infrared spectrum of the C material that HP, HT sample obtain after purifying shows 2872cm shown in Fig. 9 d/9e ^-1, 1455cm ^-1It is the C-H vibration peak of methyl, methylene; At 1724cm ^-1Near peak is the stretching vibration of C=O; At 1103cm ^-1Neighbouring is the symmetrical stretching vibration of C-O-C, shows the existence that ester is arranged.At 3400cm ^-1The stretching vibration peak that tangible O-H is arranged, showing has hydroxyl in the compound.

From ultraviolet and infrared spectrum analysis, the C material that HP, HT sample obtain after purifying is the lipid surfactant.But using the qualitative method to carbohydrate is that the anthrone reagent method is measured, and does not show the characteristic reaction of carbohydrate again.Therefore the C material should be a kind of novel lipid surfactant.

5, the interfacial tension analysis of HP, HT bacterial strain fermentation liquor

The drop volume interfacial tensimeter that utilizes German LAUDA company to produce, under 45 ℃ of current intelligences to Bacillus cercus (Bacillus cereus) HP, short bacillus (Brevibacillus brevis) HT bacterial classification at K ₂HPO ₄0.1%, NaH ₂PO ₄0.2%, (NH ₄) ₂SO ₄0.2%, CaCl ₂2H ₂O 0.001%, FeSO ₄7H ₂O 0.001%, yeast 0.1%, and, PH 7.0-7.2.Carbon source is 10% crude oil.45 ℃, respectively the nutrient solution after the 120rpm shaking table was cultivated 5 days respectively and mix and crude oil between interfacial tension detect, testing result is as shown in table 11, shows HP, the HT bacterial strain can well the metabolism biological surfactant.Biosurfactant has hydrophilic and hydrophobic grouping in the metabolite of bacterium, aligns on oil-water interfaces, improves interfacial activity, reduces interfacial tension.

The interfacial tension determination data of table 11. bacterial strain fermentation liquor

The sample title	Interfacial tension (mN/m)
		Formation water	30.24
HP	11.57
		HT	13.39
HP+HT	10.66

The composite interfacial tension experiment of embodiment 4 microbial fermentation solutions and existing ternary system

Bacterial strain: HP, HT

Culture medium A: K ₂HPO ₄0.1%, NaH ₂PO ₄0.2%, (NH ₄) ₂SO ₄0.2%, CaCl ₂2H ₂O 0.001%, FeSO ₄7H ₂O 0.001%, yeast 0.1%,, PH 7.0-7.2, carbon source is a 10-20% crude oil.45 ℃ of temperature, shaking speed: 120rpm, incubation time: 7 days.

Culture medium B:KH ₂PO ₄: 0.1-1%, Na ₂HPO ₄: 0.05-1%, NH ₄Cl:0.05-1%, NaCl:0.1-1.0%, MgSO ₄7H2O:0.01-0.5%, KCl:0.05-0.5%, FeCl ₂: 0.001-0.01%, CaCl ₂: 0.001-0.01%, MnCl ₂: 0.001-0.01%, CuCl ₂: 0.001-0.01%, ZnCl ₂: 0.001-0.01%, yeast soaks powder: 0.01-0.2%, beef extract: 0.01-0.2%, peptone: 0.01-0.2%, atoleine 0.5-2% or crude oil 1-20%, Tween80:0.01-0.2%, Tween95:0.01-0.2%, carboxylate: 0.005-0.1%, pH:6.8-7.5,121 ℃ of sterilization 15-20Min.35 ℃ of temperature, shaking speed: 120rpm, incubation time: 1 day.

Obtain zymotic fluid A and zymotic fluid B after the cultivation.

Figure 10 is the correlation curve figure of the interfacial tension of microbial fermentation solution-ternary system.Experiment condition injects activating agent (alkylbenzenesulfonate) 0.1wt% then for the microbial fermentation solution implantation concentration is 5%, NaOH 0.6wt%, the ternary prescription that polymer P (polyacrylamide) 0.1wt% forms.Experiment is the grand celebration three factory's profits that recover the oil with oily, water.

Experiment shows, is adding NaOH0.6wt%, and the ternary system of zymotic fluid preparation will be hanged down with the ternary system interfacial tension that effect crude oil interfacial tension is injected the sewage preparation during sulfosalt surfactant 0.1wt%.

Test with existing ternary prescription interfacial tension behind embodiment 5 microbial actions

Adopt embodiment 4 preparation microbial fermentation solution A to be mixed with the dilution of concentration 5%, inject oil reservoir, after 3 days, inject existing ternary system slug (surfactant sodium alkyl benzene sulfonate 0.1%, NaOH 0.6%, polyacrylamide 1000PPm), do simultaneously with reference to experiment, the result shows as shown in figure 11: microbial fermentation solution A effect crude oil rear interface tension force is starkly lower than and does not act on crude oil, and blank zymotic fluid is all very high 10 with effect front and back crude oil interfacial tension ^-1More than the mN/m.Microorganism is described in growth course, utilize the crude oil growth, and metabolism goes out biosurfactant.And rerum natura such as viscosity of crude is when improving, testing result shows, effect back acid value for crude oil can be elevated to 0.2mgKOH/g by the 0.01mgKOH/g of blank, alkali and this part contain the carboxyl material and generate salt, certain effect has also been played in reduction to interfacial tension, and therefore effect back crude oil interfacial tension will be hanged down.

Embodiment 6 bio-fermented liquids are not adding interfacial tension experiment under the artificial synthetic table agent situation alive

4 times of zymotic fluid A (embodiment 4 preparation) dilutions (this moment, its concentration was 25%) add alkali NaOH to 0.8%, polyacrylamide 1000PPm, and interfacial tension can be stablized in the time of 2 hours and remains on 10 between this moment and microbial action crude oil A ^-3MN/m.And zymotic fluid B adds alkali NaOH to 1.0% 4 times of dilutions (this moment, its concentration was 25%); With 6 times of zymotic fluid dilutions (this moment, its concentration was 16.7%), adds alkali NaOH to 0.6%, 0.8% o'clock interfacial tension and in the time of 2 hours, can stablize and remain on 10 ^-3MN/m.Interfacial tension figure sees Figure 12 a, 12b, 12c.

Simultaneously, can see from interfacial tension figure that there are some difference in the crude oil interfacial tension before and after the microbial action, this not being both because microorganism causes the metabolite of the change of crude oil property and microorganisms and ternary system compatibility.The crude oil molecular weight changes behind the microbial action, can know that from crude oil total hydrocarbon chromatogram the molecular weight after the effect reduces, and the viscosity of crude reduction, therefore the biosurfactant coupling that crude oil that this rerum natura changes and microbial metabolism produce obtains lower interfacial tension.Simultaneously, testing result shows that effect back acid value for crude oil can be elevated to 0.2KOHmg/g by the 0.01mgKOH/g of blank, and alkali and this part contain the carboxyl material and generate salt, and certain effect has also been played in the reduction of interfacial tension.

Figure 13 a, 13b, 13c, 13d are zymotic fluid A, B dilution 2-10 doubly (its change in concentration is 50%～10%), the activity figure of 30 minutes, 60 minutes and 120 minutes during alkali NaOH to 0.4%-1.2%.As can be seen from the figure, activity is better than 120 minutes in the time of 30 minutes, and it is fine much to put dynamic interfacial tension, but it is very short to hold time, and can give one's full attention to and utilize this point when living the agent compatibility with artificial synthetic table.

Embodiment 7 physical analogy oil displacement experiments

Adopt the natural core post to carry out the indoor physical simulation oil displacement experiment, verify following two prescriptions respectively.Crude oil is three factory's multi-purpose station crude oil, viscosity 19.9mPas.Microbial inoculum is the bacterial strain HT of embodiment 4 preparations, the zymotic fluid of HP.

Prescription one: the compound system of preparing with the agent alive of bacterium liquid+table behind the microbial action

Show the compound system displacement of agent preparation alive behind the microbial action with bacterium liquid+S1.Adopt this prescription mainly in order to reduce cost.

Injection mode: after the model water drive, bacterium liquid (zymotic fluid) 0.3PV of implantation concentration 5%, 45 ℃ of constant temperatures were placed 3-5 days down, injected compound system slug 0.3PV, follow-up polymer protection slug 0.2PV.Slug is formed: compound system is 2 times (50wt%) of zymotic fluid dilution+table agent 0.04%+1.0% alive alkali+2500mg/L polymer, follow-up is the 1800mg/L polymer, wherein, table agent alive is a sodium alkyl benzene sulfonate, alkali is NaOH, polymer produces polyacrylamide, molecular weight 1300-1400 ten thousand, viscosity 57.6mPas for the grand celebration auxiliary reagent factory.

Use prescription one to carry out indoor oil displacement experiment, the result is as shown in table 12, shows to improve the recovery ratio amplitude between 19.5%--21.2%, and on average improving the recovery ratio amplitude is 20.25%, suitable with independent three-component compound system displacement of reservoir oil amplitude.Use this prescription, the agent alive of expensive table only needs 0.04%, saves this expense expenditure greatly.

Table 12 prescription one experimental result

Sequence number	Oil saturation, %	Waterflood recovery efficiency factor, %	Chemical flooding recovery ratio increase rate, %,	Overall recovery factor, %	Pattern number	Perm-plug method (* 10 -3μ m 2)	Remarks
								1	68.5	42.8	20.04	64.84	69	1711
2	74.3	46.3	21.2	67.5	61	1542
								3	66.2	43.87	19.5	63.37	101	1699

Prescription two: use the displacement of existing ternary prescription behind the microbial action

Use the displacement of existing ternary prescription behind the microbial action; Adopt this prescription mainly in order further to improve recovery ratio.

Injection mode: after the model water drive, the bacterium liquid 0.3PV of implantation concentration 5%, 45 ℃ of constant temperatures were placed 3-5 days down, injected ternary system slug 0.3PV, follow-up polymer protection slug 0.2PV.Slug is formed: table agent 0.2wt%+1.0% alive alkali+2500mg/L polymer, and follow-up polymer protection slug is the 1800mg/L polymer, the agent of showing to live is an alkylbenzenesulfonate; alkali is NaOH; polymer produces polyacrylamide, molecular weight 1300-1400 ten thousand, viscosity 46.7mPas for the grand celebration auxiliary reagent factory.

Use prescription two to carry out indoor oil displacement experiment, the result is as shown in table 13, shows to improve the recovery ratio amplitude between 27.6%--30.2%, and on average improving the recovery ratio amplitude is 29%, and three-component compound system displacement of reservoir oil amplitude increases about 9% separately.

Table 13 prescription two experimental results

Sequence number	Oil saturation, %	Waterflood recovery efficiency factor, %	Chemical flooding recovery ratio increase rate, %,	Overall recovery factor, %	Pattern number	Perm-plug method (* 10 -3μ m 2)	Remarks
								1	68.2	41.4	29.2	70.6	T-51	1631
2	74.9	38.2	30.2	68.4	T-53	1891
								3	69.0	36.7	27.6	64.3	T-57	1946

The present invention is by above embodiment, illustrate utilize the microbial fermentation solution effect crude oil that HP, HT obtain after, crude oil property improves, the interfacial tension effect crude oil reduction of filling a prescription of effect back crude oil and existing ternary.Dilute 2-12 times of microbial fermentation solution and add a spot of alkylbenzenesulfonate table agent S1 (0.01wt%-0.04wt%) alive, interfacial tension can reach 10 ^-3MN/m greatly reduces cost, and has stability preferably.

The laboratory core oil displacement experiment shows that under the situation that adds a small amount of sulfonate table agent alive (0.04%), zymotic fluid dilutes 2 times, and the indoor natural core displacement of reservoir oil improves the recovery ratio amplitude on average 20.25% behind the microbial action, and is suitable with existing ternary prescription; Annotate microorganism earlier,, improve the recovery ratio amplitude, increase about 9% than independent three-component compound system displacement of reservoir oil amplitude on average 29% with the displacement of existing ternary prescription.

Claims (9)

1, a kind of microorganism-ternary composite oil-displacing method, order may further comprise the steps:

1) with Bacillus cercus (Bacillus cereus) HP, CGMCC № .1141 and short bacillus (Brevibacillus brevis) HT, at least one strain is cultivated in the culture medium that with crude oil is carbon source and is obtained fermentation among the CGMCC № .1142;

2) zymotic fluid that obtains was injected oil reservoir 3 to 7 days;

3) with the displacement of reservoir oil of ternary built system.

2, according to the described microorganism of claim 1-ternary composite oil-displacing method, it is characterized in that: culture medium is culture medium A: K in the described step 1) ₂HPO ₄0.1%, NaH ₂PO ₄0.2%, (NH ₄) ₂SO ₄0.2%, CaCl ₂2H ₂O 0.001%, FeSO ₄7H ₂O 0.001%, yeast 0.1%, and PH7.0-7.2,121 ℃ of sterilization 15-20Min, carbon source is a 10-20% crude oil; Condition of culture is 45 ℃ of temperature, shaking speed 120rpm, incubation time 7 days.

3, according to the described microorganism of claim 1-ternary composite oil-displacing method, it is characterized in that: culture medium is culture medium B:KH in the described step 1) ₂PO ₄: 0.1-1%, Na ₂HPO ₄: 0.05-1%, NH ₄Cl:0.05-1%, NaCl:0.1-1.0%, MgSO ₄7H2O:0.01-0.5%, KCl:0.05-0.5%, FeCl ₂: 0.001-0.01%, CaCl ₂: 0.001-0.01%, MnCl ₂: 0.001-0.01%, CuCl ₂: 0.001-0.01%, ZnCl ₂: 0.001-0.01%, yeast soaks powder: 0.01-0.2%, beef extract: 0.01-0.2%, peptone: 0.01-0.2%, atoleine 0.5-2% or crude oil 1-20%, Tween 80: 0.01-0.2%, tween 95:0.01-0.2%, carboxylate: 0.005-0.1%, pH:6.8-7.5,121 ℃ of sterilization 15-20Min; Condition of culture is 30～45 ℃ of temperature, shaking speed 120rpm, incubation time 1～2 day.

4, according to the arbitrary described microorganism of claim 1 to 3-ternary composite oil-displacing method, it is characterized in that: its concentration of zymotic fluid of the injection oil reservoir described step 2) is 1～5wt%, wherein contains 10 ⁷-10 ⁸Individual cell/ml.

5, according to the arbitrary described microorganism of claim 1 to 3-ternary composite oil-displacing method, it is characterized in that: in the described step 3) in the ternary built system each component be:

The zymotic fluid 10-50wt% that step 1) obtains,

Table agent 0-0.04wt% alive,

Alkali 0.4-1.2wt% and

Polymer 0.25wt%.

6, according to the arbitrary described microorganism of claim 1 to 3-ternary composite oil-displacing method, it is characterized in that: in the described step 3) in the ternary built system each component be:

The zymotic fluid 10-50wt% that step 1) obtains,

Table agent 0.2wt% alive,

Alkali 0.8-1.0wt% and

Polymer 0.25wt%.

7, a kind of microorganism-ternary composite driving finish comprises following component:

Microbial fermentation solution 10-50wt%,

Table agent 0-0.04wt% alive,

Alkali 0.4-1.2wt%, and

Polymer 0.25wt%;

Wherein, microbial fermentation solution is Bacillus cercus (Bacillus cereus) HP, CGMCC № .1141, or short bacillus (Brevibacillus brevis) HT, CGMCC № .1142 bacterial classification is cultivated the zymotic fluid that obtains in the culture medium that with crude oil is carbon source.

According to the described microorganism of claim 7-ternary composite driving finish, it is characterized in that 8, described culture medium is culture medium A: K ₂HPO ₄0.1%, NaH ₂PO ₄0.2%, (NH ₄) ₂SO ₄0.2%, CaCl ₂2H ₂O 0.001%, FeSO ₄7H ₂O0.001%, yeast 0.1%, PH7.0-7.2,121 ℃ of sterilization 15-20Min, carbon source is a 10-20% crude oil; Condition of culture is 45 ℃ of temperature, shaking speed 120rpm, incubation time 7 days.

According to the described microorganism of claim 7-ternary composite driving finish, it is characterized in that 9, described culture medium is culture medium B:KH ₂PO ₄: 0.1-1%, Na ₂HPO ₄: 0.05-1%, NH ₄Cl:0.05-1%, NaCl:0.1-1.0%, MgSO ₄7H2O:0.01-0.5%, KCl:0.05-0.5%, FeCl ₂: 0.001-0.01%, CaCl ₂: 0.001-0.01%, MnCl ₂: 0.001-0.01%, CuCl ₂: 0.001-0.01%, ZnCl ₂: 0.001-0.01%, yeast soaks powder: 0.01-0.2%, beef extract: 0.01-0.2%, peptone: 0.01-0.2%, atoleine 0.5-2% or crude oil 1-20%, Tween 80: 0.01-0.2%, tween 95:0.01-0.2%, carboxylate: 0.005-0.1%, pH:6.8-7.5,121 ℃ of sterilization 15-20Min; Condition of culture is 30～45 ℃ of temperature, shaking speed 120rpm, incubation time 1～2 day.

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